Clinical indicators do not adequately predict the long-term prognosis of patients with ST-segment elevation myocardial infarction (STEMI) following percutaneous coronary intervention (PCI). The low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio is expected to be a reliable predictor of the long-term prognosis of these patients. This study aimed to explore the correlation between the LDL/HDL ratio and long-term prognosis in STEMI patients undergoing PCI. Patients with confirmed STEMI who underwent PCI in 7 hospitals in China from January 2009 to December 2011 were enrolled. Information about clinical endpoints, including all-cause death and major adverse cardiovascular events, was collected. Overall, 915 patients were included for analysis, the average follow-up time was 112.2 months. According to the LDL/HDL ratio, the patients were divided into 3 groups using the three-quantile method: low (LDL/HDL≤1.963), medium (1.963<LDL/HDL<2.595), and high (LDL/HDL≥2.595) LDL/HDL groups. The rate of coronary revascularization was higher in the high LDL/HDL group (28.52%) than in the low  and medium  LDL/HDL groups. The hazard ratio of coronary revascularization was significantly higher in the high LDL/HDL group than in the low (P=0.007) and medium (P=0.004) LDL/HDL groups. Increased LDL/HDL ratio was an independent risk factor for long-term coronary revascularization in STEMI patients undergoing PCI . These findings suggest that an increased LDL/HDL ratio was an independent risk factor for long-term coronary revascularization in STEMI patients undergoing PCI. The risk of coronary revascularization was significantly increased in patients with LDL/HDL≥2.595. In low- and middle-income countries, mortality due to coronary heart disease will exceed that of infectious diseases by 2030, and coronary heart disease will become the disease with the highest mortality rate . ST-segmAt present, there is a lack of clinical indicators that can accurately predict the long-term prognosis of STEMI patients undergoing PCI. Commonly used methods for assessing the long-term prognosis of STEMI patients, such as coronary angiography and coronary computed tomography angiography (CTA), are mostly invasive and relatively expensive. On the other hand, biomarkers, such as serum low-density lipoprotein (LDL), high-density lipoprotein (HDL), glucose, and creatinine, are insufficient to accurately identify high-risk STEMI patients.Many studies have shown that oxidation plays an important role in the occurrence and development of coronary heart disease . Not onlIn this research, a cohort study was used to explore the correlation between the LDL/HDL ratio and the long-term prognosis of STEMI patients who underwent PCI and to clarify the predictive value of the LDL/HDL ratio for the long-term prognosis of these patients. It is expected that the results of the study will provide new clinical indicators to accurately predict the prognosis of STEMI patients, thereby promoting early identification and early intervention for high-risk STEMI patients and improving patient prognosis.http://www.chictr.org) under identifier ChiCTR-PRCH-13003570. This study was performed in accordance with the guidelines of the Declaration of Helsinki.This was a multicenter observational study involving 7 hospitals in China. Patients aged >18 years with confirmed STEMI who underwent PCI between January 2009 and December 2011 were included. The diagnosis of STEMI followed the 2007 American College of Cardiology Foundation/American Heart Association guidelines , triglycerides (TG), cystatin C (Cys C), and homocysteine (HCY) were collected within 1 hour of admission as a part of routine clinical care. All patients underwent color Doppler echocardiography within 72 h of admission.All patients had a face-to-face intensive midterm follow-up from August 2013 to January 2014 to obtain information about their clinical endpoints. In November 2019, the patients' clinical endpoints were counted by telephone and medical records.The clinical endpoints included all-cause death and major adverse cardiovascular events (MACEs). MACEs included cardiac death, coronary revascularization, nonfatal acute myocardial infarction, heart failure, and stroke. Cardiac death was defined as death without a clear non-cardiac cause; coronary revascularization was the revascularization of any disease caused by ischemic symptoms or events, including PCI and coronary artery bypass grafting (CABG). The diagnosis of nonfatal acute myocardial infarction was based on the standards proposed by the Academic Research Consortium (ARC) . The diat-test was used for comparisons between groups. Continuous variables with skewed distributions are reported as medians , and the Mann-Whitney U test was used for comparisons between groups. The cumulative hazard curve was displayed by the Kaplan-Meier (K-M) method, and the log-rank test was used for comparisons between groups. The relationship between the LDL/HDL ratio and clinical endpoints was analyzed by Cox regression analysis. Pearson correlation analysis was used to determine the correlation between the LDL/HDL ratio and Cys C and HCY. A two-tailed P value of less than 0.05 was considered statistically significant. All statistical analyses were performed using SPSS 22.0  and GraphPad Prism 7.0 .Categorical variables are reported as rates or percentages, and the chi-squared test was used for comparisons between groups. Continuous variables conforming to a normal distribution are reported as means±SD, and the independent-sampleAfter excluding 185 patients who were lost to follow-up, a total of 915 patients were included for analysis. Among them, 161 (17.60%) died, 440 (48.09%) had MACEs, and the average follow-up time was 112.2 months. According to the LDL/HDL ratio, 915 STEMI patients were divided into 3 groups using the three-quantile method: low LDL/HDL group (LDL/HDL≤1.963), medium LDL/HDL group (1.963<LDL/HDL<2.595), and high LDL/HDL group (LDL/HDL≥2.595).The comparison of baseline data among the three groups is shown inThe comparison of clinical endpoints among the three groups is shown inK-M cumulative hazard curves of clinical endpoints in the three groups are shown inThe Cox regression analysis of the LDL/HDL ratio and clinical endpoints in STEMI patients is shown inThere were weak positive correlations between the LDL/HDL ratio and both Cys C  and HCY .To our knowledge, this is the first report that the LDL/HDL ratio can predict the long-term risk of coronary revascularization in STEMI patients undergoing PCI. A high LDL/HDL ratio was an independent risk factor for coronary revascularization in STEMI patients, and the risk of coronary revascularization significantly increased when LDL/HDL≥2.595.Previous studies have shown that the LDL/HDL ratio can predict the risk of adverse cardiovascular events. After 23 years of follow-up of 2616 healthy subjects, the Kuopio Ischemic Heart Disease Risk Factor (KIHD) study found that neither LDL nor HDL had a correlation with the occurrence of sudden cardiac death (SCD) but that the LDL/HDL ratio was an independent risk factor for SCD. The risk of SCD in patients with LDL/HDL>4.22 was approximately two-fold that in patients with LDL/HDL≤2.3, suggesting that the LDL/HDL ratio is better than LDL or HDL alone for predicting the risk of SCD . A case-The exact mechanism between the LDL/HDL ratio and long-term coronary revascularization in STEMI patients has not been elucidated. Previous studies have confirmed that the occurrence of adverse cardiovascular events, including coronary revascularization, is related to biomarkers such as myeloperoxidase (MPO), glutathione peroxidase, and gamma-glutamyl transferase (GGT), which can reflect the level of human oxidation reaction -15, and2-) and hydroxyl radicals (-OH), etc., are oxygen single-electron reduction products produced in enzymatic reactions catalyzed by enzymes such as cyclooxygenase and peroxidase , including superoxide anions (Oroxidase . LDL genroxidase . In addiroxidase , advanciroxidase , and HDLroxidase . Studiesroxidase . TherefoCys C is produced by the nucleus and is released into the blood, which can be freely filtered from the glomerulus. Studies have shown that the level of Cys C is positively correlated with the infarct size of patients with acute myocardial infarction . MeanwhiIn spite of the interesting findings of the study, several shortcomings should be noted. First, the study is limited in its retrospective and observational nature. Second, the results may be confounded by familial hypercholesterolemia and other unknown or unmeasured factors. Third, the ratio of loss to follow-up in this study was relatively high, which may have introduced loss-to-follow-up bias. Finally, the last follow-up was performed via telephone and medical records, which may have also introduced bias. In our next work, we will establish a larger cohort and conduct more rigorous management to verify the prognostic value of the LDL/HDL ratio for the long-term risk of cardiovascular adverse events in STEMI patients undergoing PCI.In conclusion, our study suggests that the LDL/HDL ratio can predict the long-term risk of coronary revascularization in STEMI patients undergoing PCI. A high LDL/HDL ratio was an independent risk factor for coronary revascularization in STEMI patients. The risk of coronary revascularization in patients with LDL/HDL ≥2.595 was significantly increased.


We reviewed the records of 337 confirmed cases of tuberculosis patients in Monrovia, the capital of Liberia, 2015. The risk factors affecting the survival and multidrug-resistance of tuberculosis patients were examined. Kaplan-Meier analysis and the log-rank test were used to assess the differences in survival among the patients, while Cox regression model was used for multivariate analysis. The qualitative data was tested with chi-square test in the single factor analysis of multidrug-resistant TB. Multivariate analysis was performed using binary logistic regression analysis. The significance level for all the tests were set at 0.05. The mean period of the follow-up of patients was 10 months. In the 337 patients, 33 (9.8%) died, the 21-month survival rate was 90.2%. The results of multivariate Cox regression analysis show that overcrowding , former smoking , current smoking , multidrug-resistance tuberculosis  were risk factors for death during anti-tuberculosis treatment in TB patients in Liberia. The results of binary logistic regression analysis show that extra-pulmonary , family history of TB  and current smoking  were risk factors for multidrug-resistant tuberculosis. These results can provide insights on local tuberculosis early intervention, increase public health awareness, and strengthen the control of factors that may affect the survival and multidrug-resistance of tuberculosis patients. Tuberculosis (TB) is one of the top 10 causes of death worldwide. In 2018, 10 million people fell ill with TB worldwide and 1.5 million people lost their lives to the disease [In 2018, 87% of new TB cases occurred in the 30 high TB burdened countries .Multidrug-resistant tuberculosis (MDR-TB) is a form of TB caused by mycobacterium tuberculosis (MTB) that do not respond to isoniazid and rifampicin, the two most powerful first-line anti-TB drugs . In the In this work, we investigated the possible risk factors affecting the survival of 337 TB patients confirmed cases of tuberculosis patients in Monrovia 2015. We also estimates of the risk factors of MDR-TB patients.Using retrospective research methods, we reviewed the records of 337 patients diagnosed and treated for TB at National Leprosy and Tuberculosis Control Program in Monrovia, Liberia in 2015 . Chest XOvercrowding was defined as two or more adult of the same gender sharing the same room . TobaccoData were entered into an access database using Epidata software , cleaned and analyzed using the statistical package for the social sciences . Kaplan-Meier analysis and the log-rank test were used to assess the differences in survival among patients. Cox regression model was used for multivariate analysis. In the single factor analysis of MDR-TB, the qualitative data was tested with chi-square test, multivariate analysis was performed using binary logistic regression analysis, and the significant level for all the tests was set at 0.05.Patients in this study were fully anonymized before accessing their files. A letter was written by the Ministry of Health and Social Welfare, Liberia to the National Leprosy and Tuberculosis Control Program (NLTCP) administrator for permission to access the raw data. Before the data collection, written consent was obtained from the heads of each department of the hospital and confidentiality was maintained by coding from data collection to analysis. Each patient was assigned a unique study number for identification. Patients’ names were not used on any study related documents.Basic demographics and clinical characteristics of patients with TB are presented in p < 0.05), shown in Kaplan-Meier survival function test was used to estimate the cumulative probability of survival of different gender, marital status, religion, overcrowding, MDR-TB, occupation, CXR and smoking. The results of log-rank test show that the differences of cumulative probability of survival of various factors such as gender, marital status, overcrowding, MDR-TB, occupation, CXR and smoking are statistically significant (p < 0.05) of Kaplan-Meier univariate analyses including gender, marital status, overcrowding, MDR-TB, occupation, CXR and smoking were included in the Cox regression model for multivariate analysis. According to the multivariate analysis, former smoking , current smoking , overcrowding  and MDR-TB  were all associated with increased risk of death during TB treatment in the 337 patients .To investigate the risk factors affecting the occurrence of MDR-TB patients in Liberia, a univariate analysis was performed on TB patients admitted in 2015. As shown in p<0.05) were calculated in the binary multivariable logistic regression analysis. As shown in p = 0.017), family history of TB  and current smoking  were risk factors for MDR-TB patients.After controlling of gender and age factors, five statistically significant factors including marital status, smoking, site of TB, family history and constitutional symptom (p = 0.002) and current smoking  were all associated with increased risk of death in TB patients. The strong relation between smoking and TB were also reported in the literature [The multivariate analysis showed that former smoking . There remains limited research on the association between TB mortality rate and population residence. However, poverty, unemployment and illiteracy can affect access and the utilization of health care services and subsequently affect the outcome of TB [TB patients living in overcrowded areas were 7.942 times more likely to die than patients living in non-overcrowded areas  than males 7.8%). This finding correlates with similar studies among hospitalized pulmonary TB patients in Bolivia and Erbil which reported that after controlling for other variables, including coexisting pathology and extent of lung disease, females were found to be more likely to die from TB than males , On the %. This fp = 0.001) was identified as a risk factor for death during anti-tuberculosis treatment in TB patients. This result correlates with previous findings in Peru [MDR-TB . We also found that patients with a family history of TB were 2.387 times more likely to develop multidrug resistance . A study in Mali has confirmed that a history of contact with TB patients could significantly increase the risk of being diagnosed with MDR-TB disease [p = 0.001). Cigarette smoking adversely affects culture conversion during anti-tuberculosis treatment, which extends treatment cycles [Since MDR-TB had an important impact on patient survival compared to non-MDR-TB, we analyzed the risk of developing MDR-TB. Studies from India have reported that the prevalence of MDR-TB is higher in EPTB cases, although it has not been compared with pulmonary tuberculosis , 28. Acc disease . Patientt cycles . Prolongt cycles . ResistaIn conclusion, overcrowding, smoking, MDR-TB were important risk factors and negatively affected the survival rates of TB patients in Liberia. At the same time, extra-pulmonary, family history of TB and current smoking were significant risk factors and negatively affected the survival rates of MDR-TB patients in Liberia. The results suggested that the mycobacterium tuberculosis drug sensitivity test should be strengthened. In addition, the genotype DST method should also be considered to determine the prevalence of drug-resistant tuberculosis, so as to detect patients with MDR-TB more sensitively and early.S1 Dataset(XLS)Click here for additional data file.S1 File(DOCX)Click here for additional data file.


Background: Alterations in executive functions, emotion regulation, and their interaction are common concomitants of depression. Executive dysfunction frequently lingers after treatment, has adverse effects on daily life, and predisposes to recurrence of depression. Yet, sensitive measures of executive function for reliable assessment of cognitive outcomes are still lacking in clinical practice. To better understand the impact of depression and its most effective treatment, electroconvulsive therapy (ECT), on cognition, we assessed executive functions pre- and post-ECT and whether objective measures reflecting alterations in emotion–executive function interaction correlate with depression severity or with cognitive outcome.Methods: Executive functions were assessed in 21 patients with major depressive disorder (MDD) before and after ECT using subjective measures from the Behavior Rating Inventory of Executive Function—Adult version (BRIEF-A) and objective cognitive performance measures derived from computer-based test of executive function, Executive Reaction Time (RT) Test. In addition, we created novel indices reflecting emotional modulation of cognitive performance by subtracting different performance measures in the context of neutral distractors from those in the context of threat-related distractors. We correlated these indices with Beck Depression Inventory (BDI) and BRIEF-A scores.Results: Depression was significantly alleviated, and executive functions improved post-ECT, as seen in reduced BDI scores, BRIEF-A scores, and number of errors in Executive RT Test. Pre-ECT BDI scores correlated with threat modulation of RT (tmRT) and threat modulation of working memory (tmWM). Post-ECT tmRT correlated with several Behavioral Regulation scales and tmWM with several Metacognition scales of BRIEF-A.Conclusion: While caution is warranted, results from both subjective and objective measures suggest that ECT significantly improves executive functions and emotion regulation along with alleviation of depression. Novel indices derived from threat modulation of executive function and working memory show promise as objective biomarkers of depression severity pre-ECT and cognitive outcome post-ECT with potential for guiding depression treatments. The diagnosis of major depressive disorder (MDD) is primarily based on clinical examination and subjective evaluation of depressive symptoms with objective tests of depression lacking in the current clinical practice. Likewise, reliable and easy-to-obtain measures for cognitive outcomes of depression treatments are still missing considering the limitations of traditional neuropsychological assessments. Identifying novel biomarkers of depression severity and cognitive outcome will have broad implications in developing diagnostic tools, treatment selection, and optimizing neuromodulation treatments. Furthermore, discovery of novel biomarkers of depression will have wide impact not only on clinical practice but also on scientific endeavors aiming to gain better understanding of depression and its treatments.The most effective treatment of pharmacoresistant depression is electroconvulsive therapy (ECT) , 2. TherMDD is associated with cognitive and affective dysfunction, specifically executive dysfunction and emotion dysregulation –9. ConseExecutive dysfunction may not always be depicted with conventional neuropsychological tests, even when interfering with everyday life –20. FurtIt is crucial for survival that individuals pay attention to threat-related stimuli and allocate adequate cognitive control resources to minimize the threat, even when busy with a task engaging attention and executive functions , 25. To In this study, we studied whether the threat-related distractors impact task performance in tasks requiring multiple executive functions simultaneously, i.e., threat modulation of task performance. We assumed that patients with more severe depression would have greater impact of threat-related distractor on task performance. Threat-related stimuli may impact cognitive performance in multiple ways either impairing or improving task performance. Whether improvement or decrement is observed depends on several emotion-, individual-, and task-related factors. Negative emotional task-irrelevant stimuli may compete for the same attentional and cognitive control resources as task-relevant stimuli, leading to task interference, such as impaired response inhibition, impaired attention to global level visual features, or impaired attention to left visual field targets , 28–30. In this study, we investigated the effect of ECT on executive functions and emotional control. We hypothesized that ECT treatment will improve executive functions and, along with improved executive functions, enhance emotional control. To assess executive functions and emotional control, we used both objective and subjective measures, i.e., an experimental computer-based test of executive functions, Executive Reaction Time (RT) Test , and a cthreat modulation indices for RT (tmRT) and working memory (tmWM) by subtracting performance measures in the context of neutral distractors from corresponding performance measures in the context of threat-related distractors. These performance measures were derived from the Executive RT Test, an experimental computer-based test objectively assessing efficiency of different executive functions including working memory in the context of threat-related distractors . Thirteen subjects did not complete the study. Some subjects withdrew from the study at an early phase due to lack of sufficient mental energy to complete the experimental task and some due to interrupted ECT treatment. Fourteen participants had an International Statistical Classification of Diseases and Related Health Problems 10th revision (ICD-10) diagnosis code F33.2 (recurrent severe major depressive disorder without psychotic features) and three F32.2 (single episode major depressive disorder without psychotic features). In the final study group, the mean age of the subjects was 37.4  years.Subjects participated in two testing sessions: one just prior to the first ECT session (pre-ECT) and another after the completion of the course of ECT treatment period (post-ECT). Subjects' treatment period was 3–4 weeks, three sessions per week, and an average number of sessions was 10.9 . ECT session count varied between 4 and 21 and was guided by clinical status of the patient. Post-ECT testing was done 4–8 days after the completion of the treatment period. All patients continued their prescribed medications during ECT, and their medications were not changed during the study.ECT was administered in Tampere University Hospital Psychiatric Ward following hospital ECT protocol. ECT was applied bifrontally using a MECTA Spectrum 5000Q ECT device . Stimulation mode was a brief pulse (0.5 ms) constant current stimulation, and seizure threshold was titrated during the first session. In subsequent sessions, treatment was given at the level of 1.5 × titrated threshold. ECT energy was 81 mC for 3 patients, 157 mC for 1 patient, 162–163 mC for 11 patients, and 324–1,152 mC for 2 patients.Subjects' depression severity was assessed with Beck Depression Inventory II (BDI-II) and Montgomery–Åsberg Depression Rating Scale (MADRS) questionnaires , 43. ExeBRIEF-A questionnaire is designed for the subjective evaluation of executive functions in everyday life. It is composed of eight rating scales assessing distinct aspects of executive functions  and three composite indices  combining individual indices into higher-level composite scores.For objective assessment of executive functions, performance measures of the computer-based test of executive functions, Executive RT Test, were used. In addition, the Executive RT Test allows for the assessment of emotion–attention and emotion–executive function interaction and alterations due to neuromodulation –47 in ththreat modulation index for each subject by subtracting a cognitive measure, either reaction time or number of incorrect responses, in the context of neutral distractor from the same measure in the context of threat-related distractor, thus isolating the impact of a threat on RT (tmRT) reflecting the impact of threat on executive functions in general and on working memory (tmWM), correspondingly.Comparing performance in the context of a threat-related distractor, a line drawing of a spider, to performance in the context of an emotionally neutral control figure resembling a flower, composed of the exact same line elements as the spider, allows for assessing the impact of mere threat on attention and executive functions with the impact of a visual distractor on performance controlled for. We calculated a t-test and BRIEF-A scores and threat modulation indices with Wilcoxon signed-rank test. Reaction times in the Executive RT Test were analyzed with repeated measures ANOVA using Distractor Valence  and Test  as factors. Errors in Executive RT Test were analyzed with generalized binary logistic regression using Distractor Valence  and Test  as fixed effect predictors and Subject as random effect predictor. Each error type  had its own model, and before analysis, trial outcomes were dichotomized so that total errors outcome was “error” or “correct,” incorrect responses outcome “incorrect” or “other,” missing responses “miss” or “other,” and commission errors “commission error” or “correct.” If significant interactions were found, data were stratified into respective groups, and groups were analyzed separately.BDI and MADRS scores were analyzed using a paired Correlation analyses between subjective (BRIEF-A) and objective (Executive RT Test) measures of executive functions, depression severity , and threat modulation indices were done using Spearman rank-order correlation. As correlation analysis is sensitive to outliers , possiblp < 0.001 and BDI p < 0.001, and highly correlated, r = 0.75, p < 0.001.ECT alleviated depression significantly. Subjects' pre-ECT mean BDI score was 35.2 (SD = 7.7) and MADRS score of 36.4 (6.9), both indicating severe depression. Post-ECT mean BDI score was 17.8 (11.2), and MADRS score was 11.9 (10.5), both indicating mild depression. The changes in MADRS and BDI scores were statistically significant, MADRS F = 5.80, p = 0.028, F = 4.64, p = 0.047, F = 9.80, p = 0.006, F = 0.02, p = 0.898, RTs in the Executive RT Test were significantly slower post-ECT  than pre-ECT  . Threat- = 0.00] .The analysis of total errors, missing responses, and commission errors revealed that subjects were 37% less probable to make an error in general , more specifically, 53% less probable to miss responding in Go condition  and 72% less probable to fail in withholding a response in NoGo condition  post-ECT .The analysis of incorrect responses resulted in Test × Distractor Valence interaction. When data were stratified based on distractor valence, subjects were 59% less probable to respond incorrectly post-ECT in the context of a threat-related distractor compared to pre-ECT . There was no difference in the context of the neutral distractor . When data were stratified based on test, there were no difference between distractors pre-ECT , but post-ECT subjects were 39% less likely to respond incorrectly in the context of threat-related distractor compared to neutral distractor .p = 0.016; emotional control index (BRI) from 66.1 to 61.6, p = 0.043; and problem solving and working memory index (MI) from 74.4 to 68.6, p = 0.046. Out of the individual scales, Inhibitory control , Initiation of tasks , Flexibility to shift from one task to another , and Monitoring of one's own task execution performance  were significantly improved post-ECT. When corrected for multiple comparison, adjusted p-values—p(adj)—for GEC and Inhibit, Shift, and Initiate scales remained significant, while BRI, MI, and Task Monitoring approached significance. All scales exceptWhen pre- and post-ECT BRIEF-A T-scores  were anaSelf-Monitoring had lower post-ECT score indicating improved functioning in daily life.There were no systematic correlations between direct outcome measures of Executive RT Test and BRIEF-A before or after ECT.V = 91.5, p = 0.014. The difference between pre- and post-ECT tmRT had a trend toward significance, −5.8 ms pre-ECT and −1.7 ms post-ECT, V = 40, p = 0.089.There was a statistically significant difference in threat modulation index for incorrect responses. Threat modulation of working memory (tmWM) was +0.9% points pre-ECT, indicating that threat-related distractor impaired the working memory performance pre-ECT, and −1.1% points post-ECT, indicating that threat-related distractor improved working memory performance post-ECT. The difference was statistically significant, Correlation analysis of threat modulation indices and BDI score resulted in two significant sets of correlations. Pre-ECT tmRT and tmWM correlated with pre-ECT BDI score . These Post-ECT tmRT correlated with several behavior regulation-related BRIEF-A scales  and tmWM with several metacognition-related BRIEF-A indices . tmRT remained significant even after correction for multiple comparison see . There wWe discovered that the novel threat modulation indices isolating the impact of threat-related distractor on different cognitive performance measures correlated with depression severity pre-ECT and executive functions in daily life post-ECT. To that end, these indices show potential as biomarkers of depression severity pre-ECT and cognitive outcome post-ECT. Such biomarkers might eventually prove beneficial for objective assessment of depression severity and for optimizing cognitive outcomes of depression treatments. Furthermore, in line with previous research, subjects' depression scores were reduced from pre-ECT level indicating severe depression to a level indicating mild depression post-ECT. We also discovered that executive functions and emotion regulation improved post-ECT, as indicated with both objective (Executive RT Test) and subjective (BRIEF-A questionnaire) measures.While improved executive functions post-ECT have been previously reported , to our In large meta-analysis by Semkovska and McLoughin , impaireOne could argue that instead of true improvement in the executive functions, the results merely reflect a change in speed accuracy tradeoff , since sAs expected, ECT caused changes in subjects' emotional reactivity and emotion–cognition interaction. Threat-related distractor interfered less and even improved subjects' working memory performance post-ECT compared to pre-ECT. Working memory is known to be vulnerable to interference –70, and Correlation analysis revealed further details about the emotional reactivity. When the association between the threat modulation index and depression severity was studied, pre-ECT tmRT, and tmWM were negatively associated with depression severity . Threat-Previous depression literature suggests that depressed individuals have hyperactive subcortical structures, such as the amygdala, and hypoactive cortical control structures, such as the dorsolateral prefrontal cortex –74. On tρ > 0.7) and tmWM moderate (ρ > 0.5) correlation with several BRIEF-A scales (ρ = 0.72) with Global Executive Composite. Positive correlation between tmWM index and metacognition scales in BRIEF-A suggests that the lower the tmWM index, the less there are subjective challenges in metacognitive abilities in daily life. In other words, reduced interference or even enhancement of working memory due to threat reflects better metacognitive abilities. Negative correlation between tmRT and emotional and behavioral control scales suggests that the higher the tmRT index, the less challenges there are in behavior and emotion regulation in daily life. To that end, slowing of RTs with threat-related stimuli reflected better behavior and emotion regulation.The impact of threat on performance in Executive RT Test reflected efficiency of executive functions in daily life as measured with BRIEF-A post-ECT. More specifically, the impact of threat on RTs reflected efficiency of emotion and behavior regulation, while the impact of threat on working memory reflected overall executive functions, metacognition, and efficiency of working memory. Post-ECT correlation analysis indicated that tmRT had a strong (A scales . InteresSimilar to pre-ECT correlation with BDI, relatively more positive tmRT index indicates more “normal” reaction to threat-related distractor, now correlated with behavior and emotion regulation in daily life. Threat-related distractors compete for the same attentional resources with the task-relevant stimuli –80. LikeDespite several strengths, there are limitations in this study. This was not a clinical intervention study but rather a small experimental study, where we assessed depression and cognitive outcomes pre- and post-ECT in a treatment-resistant MDD group who received ECT for clinical reasons. Depression severity, executive functions, and emotion–executive function interaction were measured with BDI, MADRS, BRIEF-A, and performance in Executive-RT test. Thus, some of the limitations are inherently linked to small sample size and the study design, where we lack the control group and randomization of an intervention due to ethical reasons. To that end, the results should be considered preliminary, and conclusions need to be taken with appropriate caution. Especially conclusion regarding improved cognitive functions should be considered in the context of well-known ECT-mediated cognitive impairments linked particularly to memory functions .The within-subject approach used in the study could have confounding factors related to the repeated testing, most notably learning effect instead of a genuine improvement of executive functions due to ECT. However, we have previously shown the reliability of repeated assessment of executive functions with Executive RT Test when initial learning is accounted for , and thoRegarding limitations of the threat modulation indices, tmRT and tmWM, as depression biomarkers, there was a lack of correlation with MADRS pre-ECT and with BDI and MADRS post-ECT. While both BDI and MADRS assess depression severity, they differ in a critical way, as BDI is a self-report measure of the depression severity unlike clinician-rated MADRS. The fact that MADRS lacks a structured interview may affect reliability and consequently contribute to lack of correlation with the threat modulation indices pre-ECT. Lack of correlation post-ECT may relate either to the lack of sensitivity of these indices in mild depression, lack of adequate variance in mild depression scores post-ECT, or to a small sample size. Another reason for lack of correlation of threat modulation indices with depression metrics post-ECT could relate to habituation to threat-related stimuli. We did not evaluate subjective perception of the threat nor unpleasantness of threat-related stimuli in this study. The threat-related stimuli were task-irrelevant small black line drawings of spiders presented in the middle of a visually significantly more salient task-relevant stimulus, large and colorful traffic light. Thus, selective attention was not voluntarily geared to emotional distractors. Distractor stimuli were also presented very briefly, only for 150 ms, and many of the subjects were not even aware of distractors while focusing on cognitively challenging task. It is likely that biologically relevant prototypical stimuli engage rapid and evolutionarily hard-wired pathways directly from the thalamus to the amygdala, making the effect of threat stimuli rapid and rather robust . The effIn the future, to further investigate and develop biomarkers of depression based on the impact of threat on cognitive performance, it will be critical to study habituation to threat stimuli and weather potential habituation depends on efficiency of frontal control circuits or depression severity. However, the fact that threat-related stimuli continued to have an impact on cognitive performance post-ECT and that tmRT correlated with several behavior regulation related BRIEF-A scales and tmWM with several metacognition related BRIEF-A indices suggests that subjects did not habituate to these stimuli, at least to the extent where threat would not have any impact on their performance.Another potential confounding factor in the study is the possibility that the study group was not a representative sample of the typical patients receiving ECT for refractory MDD. There are some challenges in the recruitment of subjects with severe MDD to participate in a study, which requires significant cognitive effort. Volunteering as a subject in a research study requires cognitive energy and initiation abilities, which are frequently compromised in MDD. Many subjects withdrew their consent during the first test session due to lack of sufficient energy for carrying out the study task. Thus, there may have been an unintentional bias in the study group favoring subjects with adequate level of mental energy over those with severe lack of energy.In summary, these preliminary results from a small study with aforementioned limitations suggest alleviated depression and improved executive functions and emotion regulation 4–8 days post-ECT. Subjects were less impulsive and more accurate in computerized test of executive functions and experienced improved executive functions in daily life. Furthermore, their reactions to emotional distractors were normalized, and their working memory better shielded from emotional interference. Threat modulation index correlated with depression severity pre-ECT and with BRIEF-A questionnaire scores post-ECT. The fact that threat modulation indices reflect the level of depression in more severe depression and level of executive functions and emotion regulation in milder depression may highlight how intricately intertwined these phenomena are. In milder depression, threat-modulation biomarker might reflect the resilience or susceptibility to reoccurrence of depression. This, however, requires further studies to be confirmed.The current study presented novel threat modulation indices that may serve as biomarkers for depression severity pre-ECT in more severely depressed population and as indicators of emotion regulation and efficiency of executive functions in milder depression post-ECT. In addition to potentially contributing to improved diagnostic tools, such biomarkers have the potential and applicability in scientific endeavors aiming at better understanding the role of executive functions and emotion regulationin depression.Restrictions apply to the datasets. The datasets for this article are not publicly available because the written consent forms signed by the participants did not provide information concerning public distribution of data.The studies involving human participants were reviewed and approved by Tampere University Hospital Ethical board. The patients/participants provided their written informed consent to participate in this study.JP was the main author of the manuscript, participated in data collection, data analysis, interpretation of the results, and authoring of the manuscript. KJ was responsible for the recruitment of subjects, ECT as part of subjects' clinical treatment and participated in interpreting the results, and authoring the manuscript. PH was responsible for data collection and participated in authoring the manuscript. JC participated in interpretation of the results and authoring of the manuscript. KH was responsible for the experimental design, invented the novel biomarkers based on threat-modulation of cognitive performance, supervised data collection and data analysis, participated in the interpretation of the results, and authoring of the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Background/Objectives: Aerobic exercise and mind-body exercise, are vital for improving motor and non-motor functional performance of Parkinson’s disease (PD). However, evidence-based recommendations on which type of exercise is most suitable for each individual are still lacking. Therefore, we conduct a network meta-analysis to assess the relative efficacy of aerobic and mind-body exercise on motor function and non-motor symptoms in Parkinson’s disease and to determine which of these therapies are the most suitable.Design: A network meta-analysis and dose-response analysis.Setting and Participants: Medline, Embase , and the Cochrane Central Register of Controlled Trials were comprehensively searched for related trials through April 2021.Measurements: Study quality was evaluated using the Cochrane Risk of Bias Tool. The effect sizes of continuous outcomes were calculated using mean differences (MDs) or standardized mean differences (SMDs). A network meta-analysis with a frequentist approach was conducted to estimate the efficacy and probability rankings of the therapies. The dose-response relationship was determined based on metaregression and SUCRA.Results: Fifty-two trials with 1971 patients evaluating six different therapies were identified. For the UPDRS-motor score and TUG score, yoga all ranked highest . The SUCRA indicated that walking may best improve the BBS score (SUCRA = 90.2%). Depression, cognitive and activities of daily living scores were significantly improved by yoga . In the dose-response analysis, 60-min sessions, two times a week might be the most suitable dose of yoga for reducing the UPDRS-motor score of PD patients.Conclusion: Yoga and walking are important options for increasing functional mobility and balance function, and yoga might be particularly effective for decreasing depressive symptoms and cognitive impairment and improving activities of daily living in PD. The potential optimal dose of yoga for enhancing motor ability in PD patients is 60-min sessions, two times a week.Registration: PROSPERO CRD42021224823. According to epidemiological studies of Parkinson’s disease (PD), the global PD population reached approximately 6,100,000 in 2016 , 2019, mAlthough pharmacotherapy is currently still the first-line treatment, side effects and response fluctuations limit its application . The latNetwork meta-analysis is a technique that uses direct and indirect results to compare multiple interventions simultaneously and estimates the rank order of the treatments, providing evidence-based recommendations for assisting medical decision making . TherefoThis network-meta analysis was prospectively registered in the International Prospective Register of Systematic Reviews (PROSPERO) with a record number CRD42021224823.The electronic databases Medline, Embase , and the Cochrane Library were searched for all relevant citations published from inception through April 3, 2020. We used various combinations of medical subject headings and free terms, which included Parkinson’s disease, aerobic and mind-body exercise (a list of relevant exercise interventions), and randomized clinical trial designs (search strategies are listed in Eligible studies were included when they met the following criteria: (1). All studies were randomized controlled trials (RCTs). (2) The population of the included studies was adult patients diagnosed with PD. (3) Interventions contained at least one of the following exercises: treadmill exercise, walking, cycling, dance, yoga, and tai chi. The control group received usual care, was a waitlist control or performed other non-aerobic and non-mind-body exercise. (4) Outcomes of the studies included at least one of the following measurements: Unified Parkinson’s Disease Rating Scale (UPDRS) score [UPDRS or Movement Disorder Society-Unified Parkinson’s disease rating scale scores (MDS-UPDRS)], Berg Balance Scale (BBS) score, Timed-Up-and-Go (TUG) score, psychosocial outcomes  and activities of daily living (ADL) .Studies were excluded if insufficient data or information concerning assessment was provided. We also excluded quasi-RCTs, animal trials, clinical protocols, conference abstracts, case reports and systematic reviews.Two investigators (YX & HG) independently extracted the relevant data and information from the eligible studies. Basic information about the study characteristics , population, interventions and comparisons , and measurement outcomes was extracted. Another two reviewers (CW & CT) evaluated the quality of the included studies based on the standard criteria of the Cochrane Risk of Bias Tool . Any disThe outcomes of this network meta-analysis were listed as follow:Motor outcomes: UPDRS-motor score ; BBS anNon-motor outcomes: Psychosocial outcomes  and ADL.Safety outcomes: Non-serious and serious adverse events.I2 statistic. If statistically quantifiable heterogeneity existed, a random-effects model was fitted. Otherwise, we used a fixed-effect model . Mean differences  or standardized mean differences  with 95% confidence interval (CI) was reported as a continuous outcome . We assect model .p-value of the design-by-treatment interaction model and the network side-split analysis exceeded 5%, a consistency model was used to evaluate the effect size of the multiple treatment comparisons.Then, we conducted a network meta-analysis for each outcome using a frequentist approach and the netmeta command in Stata version 14 . We quanWe estimated the ranking probabilities of each treatment for different outcomes based on the surface under the cumulative ranking curve (SUCRA) and mean ranking . MoreoveWe retrieved a total of 5,919 publications from the electronic databases and eliminated 1,592 duplicate publications. A total of 4,327 publications were left for screening according to their titles and abstracts. Of those, 4,095 publications were removed, and 232 remaining publications were then identified as potentially eligible studies and underwent a full-text review. Finally, fifty-two relevant publications were included in the network meta-analysis .A total of 52 eligible RCTs with 1,971 patients diagnosed with PD were included in this network meta-analysis. The interventions of the included trials mainly included treadmill exercise, walking, cycling, dance, yoga, and tai chi. Most trials used usual care, waitlist or other non-aerobic exercise as the control. The duration of each intervention varied from 20 to 90 min per session, and the frequency ranged from one to five times per week. Forty-two RCTs assessed motor function using the UPDRS-motor score. Fourteen studies used the BBS to assess balance function. Twenty-three studies evaluated mobility function using the TUG test, twelve RCTs assessed depressive outcomes, 13 RCTs evaluated cognition, and 17 studies reported ADL outcomes see .Among the 52 RCTs, 65.38% had a low risk in terms of random sequence generation, and 63.46% reported the use of allocation concealment methods. Few studies (11.54%) used blinding methods for participants and personnel because exercise is a non-pharmacologic treatment. Forty studies (77.36%) reported a low risk for bias in terms of blinding the outcome assessment. RCTs (84.62%) had a low risk of attrition bias. Forty-two studies reported a low risk of reporting bias. Overall, 67.31% were deemed to have a low risk of poor methodological quality, whereas 17 studies were regarded as having poor methodological quality .Forty-two RCTs with six different therapy categories assessed motor function using UPDRS III-motor measurements. Treadmill exercise contributed 18.8% to the network plot, walking 8.2%, cycling 4.7%, dance 9.4%, yoga 5.8%, tai chi 7%, and the control group 45.8% .The pairwise meta-analysis illustrated that all therapies could decrease the overall UPDRS motor score. Those performing treadmill exercise, walking, dance, yoga, or tai chi all functioned better than those in the control group in terms of the UPDRS-motor score .p-values exceeded 0.05 ], walking , dance , yoga , and tai chi  were superior to the control in reducing the UPDRS-motor score. Yoga and walking were significantly more effective in decreasing UPDRS-motor scores than treadmill therapy . The ranp = 0.15 > 0.05, Because yoga was ranked first for the UPDRS-motor score, we further analyzed the optimal dose of yoga therapy for improving motor function in PD patients. We analyzed the potential optimal time of yoga per session in UPDRS motor score. Our metaregression revealed that the effect size was not significantly increased with increasing yoga intervention time each session , tai chi , and treadmill exercise  were more beneficial in reducing the TUG score than the control . MoreoveFourteen studies including six treatments were included in the network meta-analysis evaluating BBS outcomes . Figure p > 0.05, The comparisons indicated satisfactory consistency  to comprehensively evaluate the most suitable treatment for improving motor function in PD. The results showed that the yoga, walking, dance, and tai chi groups had higher SUCRA values in the clustered ranking plot, which indicated that yoga and walking in particular, in addition to dance and tai chi, are the most suitable therapies for increasing overall motor function in PD patients .Twelve studies with five treatments assessed depression scales . The paip-values exceeded 0.05 in the network side-split model, which revealed that the direct and indirect comparison outcomes were consistent ]. Yoga, treadmill exercise and dance were significantly superior to cycling for reducing depression scores. Yoga therapy (SUCRA: 86.3%) was ranked highest for ameliorating PD with depression, followed by walking, treadmill exercise and dance .Thirteen studies with five interventions assessed cognitive function . The paip-values of consistency tests were higher than 0.05, which indicated that direct and indirect effects had good consistency ]. The ranks of the five interventions for enhancing cognitive function in PD were as follows: yoga, dance, treadmill exercise, walking, and cycling .Seventeen studies assessed ADL in PD. We conducted a pairwise meta and the results demonstrated that treadmill exercise, cycling and yoga were all associated with significantly greater changes in ADL scores than the control .p–values of consistency tests were substantially higher than 0.05, which demonstrated that the network analysis had good consistency ; SMD = –0.53 ; and SMD = –0.47 , respectively]. Furthermore, yoga and treadmill therapy were significantly more effective in improving performance of activities of daily living than dance. The rankings of these five interventions, which were based on the SUCRAs for the ADL scores, showed that yoga ranked first, followed by cycling, treadmill exercise and tai chi .Thirty-three studies reported adverse events related to exercise therapies. Among these RCTs, 21 studies reported that no adverse events occurred during treatment. The remaining studies reported a minimal number of adverse events, mainly knee, neck and back pain, muscle soreness and non-injurious falls. All these events and the symptoms were resolved by resting or simple treatment without further management see .In order to minimize the influence of heterogeneous baseline severity of PD in our study, we restricted our analysis into trials with early to-moderate stage of Parkinson’s disease as sensitivity analysis. The result showed that five therapies were superior to the control in reducing the UPDRS-motor score and yoga ranked first based on the SUCRA (93.4%). Besides, a sensitivity analysis after excluding studies that contained resistance active exercise as control group was conducted. All therapies except cycling were significantly superior to placebo and yoga also ranked first based on the SUCRA (95.2%), which were consistent with those previous produced . BesidesWe constructed a funnel plot of our outcomes to assess publication bias. The plots all showed that the effect size of the included studies was distributed symmetrically, which demonstrated that there was minimal publication bias in this analysis .We conducted a network meta-analysis that included 52 RCTs with six different treatments and obtained several principal findings summarized as follows.The pooled results suggested that yoga, walking, dance, and tai chi were superior in reducing the UPDRS-motor scores of PD patients, and yoga and walking might be highly effective based on the SUCRA rankings. In the TUG test, yoga, tai chi and treadmill exercise were superior to the control. Additionally, yoga ranked first in decreasing the TUG score. Moreover, in terms of the BBS Walking had the highest efficacy among these various treatments in terms of BBS score. According to the clustered ranking plot of the network, yoga and walking might be the important therapies for comprehensively increasing motor and balance function in PD patients. Previous studies also found that yoga and walking exerted beneficial effects on motor function in PD .Regarding psychosocial outcomes, the results suggested that yoga and dance could be significantly more effective in reducing depressive symptoms and improving cognitive function. Overall, yoga might be the important alternative therapy for PD patients with depression or dementia according to the SUCRA rankings. These findings were in line with previous studies revealing that patients report that yoga is enjoyable, feasible and beneficial regarding their depressive and anxiety symptoms . In addiFor the ADL scale, we found that yoga, cycling, and treadmills were more effective than controls in improving the ADL of PD patients, especially yoga, which ranked higher in the ADL assessment, suggesting that yoga might be the vital option for patients with poor activities of daily living. A previous review also agreed with our results and indicated that yoga therapy improved the PDQ-ADL score and mood of PD patients more than other therapies .No severe adverse events occurred during exercise therapies. Only a few patients suffered muscle soreness or minor low back or knee pain. All these events can be resolved by resting and not affect the treatment process. Overall, aerobic or mind-body therapy is a safer treatment for addressing PD symptoms than other therapies.With regard to the quality of the included RCTs, 67.31% were determined to be high-quality RCTs, which indicated that the pooled results were robust and reliable overall. However, the high risk of improper blinding of participants still existed in this analysis, which might decrease the strength of the evidence to a degree.To our knowledge, this network meta-analysis is the first study to include all previously examined types of aerobic and mind-body exercise and to explore the rankings of various types of therapies for PD according to a comprehensive range of outcomes. Most of the previous meta-analyses focused on evaluating the efficacy of only one type of exercise for PD . HoweverOverall, our results have several clinical implications. First, the results suggest that yoga and walking may be the important exercises for enhancing motor function and balance functional ability. Second, yoga as a mind-body exercise is a good option for PD complicated with depressive disorder and cognitive impairment. Moreover, yoga therapy for 60 min, two times a week can be recommended as the potential optimal dose for improving mobility in PD. Therefore, these evidence-based results could be recommended for PD patients and assist clinical doctors in making appropriate decisions.Our analysis has several limitations. In terms of non-motor outcomes, few studies and interventions were included in the analysis. Moreover, RCTs of head-to-head comparisons with other active therapies that included non-motor assessments were lacking, providing little direct evidence for overall or accurate evaluation of the efficacy of exercise therapies for non-motor symptoms  in PD. Besides, because of the limit number of yoga studies and the limitations of SUCRA, the results in this study should be interpreted with caution. Hence, more studies involving yoga are needed. In addition, most of the included studies lacked participant blinding because the exercise therapies were non-drug interventions. However, this could have resulted in bias for the intervention group.The network meta-analysis reported herein indicates that aerobic and mind-body therapies significantly improve the motor function of PD patients at all stages. Yoga and walking could be prior to recommend for PD patients with motor symptoms and balance impairment. Yoga may also be the important option for decreasing depressive disorder and cognitive impairment and could be recommended as part of treatment for life style changes in patients with PD. The potential appropriate dose of yoga for enhancing motor ability in PD patients is 60-min sessions, 2 times a week.The original contributions presented in the study are included in the article/MZ and DC designed the study and revised the manuscript for important intellectual content. YX and HG acquired the data. CW and CT analyzed and interpreted the data. CW drafted the manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Single-cell RNA sequencing (scRNA-seq) is a tool for studying gene expression at the single-cell level that has been widely used due to its unprecedented high resolution. In the present review, we outline the preparation process and sequencing platforms for the scRNA-seq analysis of solid tumor specimens and discuss the main steps and methods used during data analysis, including quality control, batch-effect correction, normalization, cell cycle phase assignment, clustering, cell trajectory and pseudo-time reconstruction, differential expression analysis and gene set enrichment analysis, as well as gene regulatory network inference. Traditional bulk RNA sequencing does not address the heterogeneity within and between tumors, and since the development of the first scRNA-seq technique, this approach has been widely used in cancer research to better understand cancer cell biology and pathogenetic mechanisms. ScRNA-seq has been of great significance for the development of targeted therapy and immunotherapy. In the second part of this review, we focus on the application of scRNA-seq in solid tumors, and summarize the findings and achievements in tumor research afforded by its use. ScRNA-seq holds promise for improving our understanding of the molecular characteristics of cancer, and potentially contributing to improved diagnosis, prognosis, and therapeutics.The online version contains supplementary material available at 10.1186/s13046-021-01955-1. Tumors are generally considered to be of monoclonal origin, with mutations facilitating the expansion of a malignant cell in the body to visible tumor tissue, as well as from carcinoma in situ to metastatic carcinoma. Notably, patient-derived tumor tissue includes cancer cells as well as other cell types, such as infiltrating immune cells and fibroblasts. Therefore, the use of traditional bulk RNA sequencing technology can only permit an average qualitative characterization of such highly complex tissue, representing the average of cancer cells and non-cancer cells. Although bulk RNA approaches have made many invaluable contributions to medical science , such apSince the first single-cell RNA sequencing (scRNA-seq) study was published in 2009 , compariIn the present review, we introduce methods for the preparation of solid tumor samples, related scRNA-seq platforms, the analysis of scRNA-seq data, and achievements in tumor research facilitated by the use of scRNA-seq.To carry out scRNA-seq, solid tumor samples first need to be processed to effectively isolate viable single cells from the tissue of interest . Thereaf3 and washed with PBS to remove fat, visible vessels, and surrounding necrotic areas [Many published articles on the use of scRNA-seq in cancer research have detailed the preparation of solid tumor specimens. After cancer is diagnosed, tumor tissues are removed by biopsy and treated immediately to preserve single cells. Tumor tissue is usually cut into sections of approximately 1 mmic areas . Tissue ic areas , using aic areas , 9, or tCurrently, major single-cell sequencing platforms include the 10X Genomics Chromium, Nadia (Dolomite Bio), Illumina Bio-Rad ddSEQ Single-Cell Isolator, BD Rhapsody Single-Cell Analysis System (BD), ICELL8 Single-cell System (Takara), Fluidigm C1, and others. Although new platforms for scRNA-seq are still being developed, the most widely used platform remains the 10X Genomics Chromium.Selecting the most suitable sequencing platform for your research project is a key step for achieving the desired research results. There are a number of published articles comparing performance, cost, and other aspects of existing platforms. In 2019, Zhang et al.  comparedData generated during scRNA-seq usually gets processed via two analytical procedures: pre-processing  and downstream analysis . The following section will briefly introduce the significance of and methods for these analytical procedures.Quality control should always be carried out as the first step for scRNA-seq data. Various methods for quality control have been developed –14. DataDuring the scRNA-seq experimental procedure, when cells subject to different conditions are cultured, captured, and sequenced separately, batch effects will be evident . There aData normalization is essential for scRNA-seq to make gene expression comparable within and/or between samples. A number of methods have been developed for the normalization of RNA-seq data –33. HoweDetermining the cell cycle phase of a single cell can facilitate understanding of biological processes such as tumorigenesis –41 and cOne of the basic goals of scRNA-seq data analysis is to identify cell types from experimental samples to elucidate tissue complexity and heterogeneity. Due to the importance of cell type recognition, efforts have been made to develop new algorithms, including CountClust , CIDR 4, SIMLR , SAFE 5, and othBy reconstructing cell trajectory and pseudo-time based on scRNA-seq data, dynamic processes in cells can be calculated and simulated, which is of great significance for understanding the transition between cell states in cancer . CurrentOne of the most common uses of gene expression data is for the identification of differentially expressed (DE) genes under different experimental conditions , and thus to determine the root cause of phenotypic differences observed under different conditions . A zero We usually group DE genes according to their participation in common biological processes to facilitate the interpretation of results . ExistinThe combination of active transcription factors and their target genes is usually described in gene regulatory networks (GRNs). Revealing these regulatory interactions is the goal of GRN inference methods, providing valuable insights for the identification of causal regulatory factors in biological processes . A classCancer patients may be unresponsive to therapy due to drug resistance and metastasis of single cells, both of which constitute major challenges in the treatment of malignant tumors. About 90% of available drugs are effective in less than half of patients . Cancer The considerable heterogeneity of tumors and tumor tissue samples between different patients is an important reason for treatment failure . TherefoGlioblastoma (GBM) is the most common primary malignant brain tumor in adults . It is tMelanoma is a highly malignant skin cancer with four clinically distinguishable subtypes and is responsible for approximately half of skin cancer-related deaths in Japan . Gerber Head and neck squamous cell carcinoma (HNSCC) encompasses a group of malignant tumors originating from the squamous epithelium of the oral cavity, oropharynx, larynx, and hypopharynx –98. PuraLung cancer (LC) can be divided into two subtypes: small-cell lung carcinoma (SCLC) and non-small-cell lung carcinoma (NSCLC), among which NSCLC accounts for the majority of LC cases . LambrecAcute myeloid leukemia (AML) is a highly heterogeneous hematological malignancy. Van Galen et al.  profiledThe ability of single-cell gene expression profiling to identify specific patterns of gene expression allows for the elucidation of mechanisms underlying tumor invasion and metastasis –105 thatPancreatic cancer is one of the leading causes of cancer-related death in developed countries, and it ranks 7th in cancer-related deaths among both men and women . Ting etBreast cancer is by far the most common malignancy in women  as well When anti-cancer drugs are used to target tumor cells, some cells develop resistance to drugs and contribute to disease relapse, rendering treatment ineffective. ScRNA-seq can be employed to distinguish between primary and recurrent tumor cells, providing insight into the cell subsets responsible for tumor recurrence, mutated genes, and multiple pathways that drive tumor growth. Thus, scRNA-seq can guide strategies for the treatment of tumors  is one of the most common malignant brain tumors in children, with a higher incidence in children aged 3 to 4 years and 8 to 10 years, and is slightly more prevalent in boys , with noVarious types of immune cells infiltrate tumor tissue –128. TheLYAN was highly expressed in activated CD8+ T cell and Tregs, which could inhibit the function of CD8+ T cells. In 2017, Muller et al. [Hepatocellular carcinoma (HCC) usually occurs in the context of advanced chronic liver disease, and is primarily associated with hepatitis B or C virus infection and alcohol abuse, accounting for 70–85% of the total cases of liver cancer . Zheng er et al.  applied r et al.  used pair et al.  performeColorectal cancer (CRC) is a common gastrointestinal malignancy, and is the third most common cancer in the United States . Zhang eScRNA-seq can solve problems that previous technologies could not address, thus resolving debates and contributing to the development of oncology.The relationship between human cytomegalovirus (HCMV) and GBM is an ongoing debate, as the detection of HCMV in GBM samples via molecular assays may be influenced by cellular heterogeneity. In 2017, Johnson et al.  aligned ScRNA-seq can be used to better understand the occurrence and development of tumors at the cellular level, which enables to the exploration of new therapeutic methods , and proved that G34R/V HGGs originated from interneuron progenitors expressing GSX2/DLX using scRNA-seq, and then confirmed this result using epigenomic profiles.In 2019, Hovestadt et al.  obtainedAt present, practice has proven the guiding significance of scRNA-seq in anticancer protocol design, with important clinical application value in personalized diagnosis, treatment, and prognosis, as well as the evaluation of treatment effect for highly heterogeneous tumors  are diagnosed with metastases, and metastatic renal cell carcinoma (MRCC) is one of the most drug-resistant malignancies . In 2016Modern cancer treatment approaches are often limited in their efficacy. In-depth analysis of malignant proliferative cells and immune cells at single-cell resolution is conducive to the identification of potential therapeutic targets, contributing to the development of new drug therapy and the improvement of patient survival rate  is an epithelial carcinoma arising from the lining of nasopharyngeal mucosa –152. AltHigh-throughput scRNA-seq technology is transforming biomedical research by establishing the transcriptome profiles of individual cells from different tissue samples , 153. WhIn time, scRNA-seq will be more widely used in the study of various tumor types, helping to develop better diagnostic and prognostic biomarkers, and design more accurate anticancer therapy to improve therapeutic efficacy and avoid tumor drug resistance. While the transcriptome of cells can be studied through scRNA-seq, there are some limitations to this method, particularly with regard to genomic and protein-level information. When CITE-seq was used in combination with scRNA-seq, the transcripts and surface marker proteins of cells could be detected simultaneously, and the phenotypes that could not be detected by scRNA-seq alone could be found . The comAdditional file 1 : Fig. 1. Application of scRNA-seq in tumor research. ScRNA-seq can be used to analyze the proportion of various immune cell types and immune repertoire characteristics, guiding choices related to immunotherapy. The analysis of single tumor cells can improve our understanding of drug resistance, invasion, and metastasis mechanisms, as well as cell origin and evolution, in turn aiding the choice of targeted drugs. T, T cell; B, B cell; NK, natural killer cell; TAM, tumor-associated macrophage; Th, helper T cell; Tc, cytotoxic T cell; Treg, regulatory T cell; Tm, memory T cell; Ts, suppressor T cell; B1, T cell independent B cell; B2, T cell dependent B cell.Additional file 2 : Table 1. Overview of related studies using scRNA-seq.Additional file 3 : Table 2. Overview of related articles using scRNA-seq.Additional file 4 : Table 3. Overview of related studies using scRNA-seq.Additional file 5 : Table 4. Overview of related articles using scRNA-seq.Additional file 6 : Table 5. Overview of related articles using scRNA-seq.Additional file 7 : Table 6. Overview of related articles using scRNA-seq.Additional file 8 : Table 7. Overview of related articles using scRNA-seq.


In 2040, the older population's growth rate in sub-Saharan Africa (SSA) will be faster than those experienced by developed nations since 1950. In preparation for this growth, the National Institute on Aging commissioned the National Academies' Committee on Population to organize a workshop on advancing aging research in Africa. This meeting provided a platform for discussing some areas requiring improvement in aging research in SSA regions. We believed that conducting a systematic review of peer-reviewed articles to set priorities for aging research in SSA is warranted. Therefore, this article is the first in a Four-Part series that summaries the types and trends of peer-reviewed studies in SSA.Search-Appraisal-Synthesis-Analysis Framework. We systematically searched multiple databases from inception till February 2021 and included peer-reviewed articles conducted with/for older adults residing in SSA. Conventional content analysis was employed to categorize studies into subject-related areas.This systematic mapping review followed the We included 512 studies . Studies were conducted in 32 countries. Quantitative studies included were observational studies: cross-sectional , longitudinal , and case-control ; and experimental studies: pre-post design , randomized control trial ; and not reported . Fifteen qualitative studies did not state their study design; where stated, study design ranged from descriptive , ethnography , grounded theory , narrative , phenomenology , interpretative exploratory , case studies . Of the 15 mixed-method studies, seven did not state their mixed-method design. Where stated, design includes concurrent (n = 1), convergent (n = 1), cross-sectional (n = 3), informative (n = 1), sequential exploratory (n = 1) and retrospective (n = 2). Studies were classified into 30 (for quantitative studies) and seven  subject-related areas. HIV/AIDs-related and non-communicable diseases-related studies were the most predominant subject-related areas. No studies explored the transdisciplinary co-production of interventions.There are glaring gaps in ageing research in SSA, especially mixed-methods and RCTs. A large number of studies focused on HIV/AIDs and non-communicable disease-related studies. National and international funding agencies should set up priority funding competitions for transdisciplinary collaborations in ageing research. With increasing life expectancy, older people’s population is projected to grow worldwide, including the world's developing regions . In 1990Although the population of the older adults in the SSA seems not to be the primary concern of several member nations in this region, evidence has demonstrated the impact of these growing populations on the country's economy, health care system, educational system and even workforce . The labThe United Nations has advocated for more evidence/data to understand the status and the needs of older persons in the SSA. For instance, in 2004, the National Institute on Aging commissioned the National Academies’ Committee on Population to organize a workshop on advancing ageing research in Africa . This eiThe World Health Surveys and the follow-up on Global Aging (SAGE) through its Wave I & II has improved data availability on health and related social issues of ageing in three SSA countries: Ghana, South-Africa, and Uganda . There aPart I summarises the types and trends of ageing research in SSA and provided specific recommendations regarding the types of ageing research in SSA.Part II will report the quality of the included studies .Part III will review the longitudinal studies of Ageing in Africa and describe the impact of longitudinal studies in Ageing on the ageing research in SSA.Part IV will present a set of ageing research priorities in SSA developed by stakeholders  through a modified Delphi process.This paper: Part 1 summarises (a) the types and trends of ageing research in SSA and (b) provides updated specific recommendations for ageing research in SSA.Search-Appraisal-Synthesis-Analysis Framework (SALSA) , and experimental studies [pre-post design (n = 4) -26; rand, 30%, anThree-quarter of the included articles  provided statements of obtaining ethical approval for their study or that ethical approval is not required, while 113 (27%) articles did not provide this statement. More than half of the included articles  provided statements of obtaining informed consent from the participants before commencing with data collection.Our content-coding resulted in 30 subject-related areas. Most studies included were categorised into Non-communicable disease-related studies (n = 41), HIV-related studies (n = 30), Physical functioning-related studies (n = 26), Cancer-related studies (n = 28), quality of life/well-being related studies (n = 22), Dementia/cognitive impairment-related studies (n = 30), Neurological-related studies (n = 15). See Of the 71 included qualitative articles, 38 (54%) and 33 46%) principal/corresponding authors' affiliations were health sciences and were social science, respectively, and one was humanities. The sample size ranged from 1 ,40 to 21% principOf the 15 mixed-method studies, the principal/corresponding author's affiliation was in health sciences n = 13, 85%) and social sciences . Participants were recruited from the hospital , and community ; in rural , urban , both  settings and not described in one study . Six stu% and socThe themes from the qualitative and mixed-method studies were group into seven subject related areas. They include health/social care experience (n = 24), HIV/AIDs related studies (n = 21), contemporary studies (n = 14), older peoples’ care-related studies (n = 14), loneliness-related studies (n = 5), dementia care-related studies (n = 4) and quality of life/physical functioning related studies (n = 4). See This mapping review summarized the types and trends of research on ageing in the SSA region. Included studies were predominantly quantitative-cross-sectional studies, published in the last ten years and mainly in South Africa, with HIV-related studies and non-communicable disease-related studies being the most predominant studies on ageing in SSA. These studies published mainly in the social and health science paradigm increased over the last ten years could indicate interest in researching to improve older adults' health and social life in SSA. We could not find any empirical evidence to support or refute if ageing research has increased in the Latin American and Caribbean countries with a similar projection of the ageing population with the SSA countries.Almost half of the included studies were conducted for or with older adults residing in South-Africa. This finding highlights the underrepresentation of other SSA countries, indicating a dearth of ageing research in SSA countries outside South Africa. This finding agrees with the previous review exploring end-of-life research in SSA, reporting a lack of socio-cultural aspects of end-of-life care in African countries outside South-Africa . This hiAside from systematic reviews and meta-analysis, randomized control trials (RCT) are the highest level of evidence for clinical decision-making . Two perThere are fewer qualitative studies compared to quantitative studies included in this review. This is not surprising because historically, quantitative studies are often valued than qualitative studies , maybe bThe mixed-method was the least studied method used in ageing research in SSA. Although 64% of researchers reported the type of mixed-method design , they failed to demonstrate when the qualitative and quantitative data were mixed; maybe because of the many journals' word counts limit. Mixed-methodology researchers had advocated that researchers who intend to use a mixed-method as their research method must demonstrate when and how the data was mixed . The mixHIV-related studies and non-communicable disease (NCDs)-related studies were predominant. With the advent of antiretroviral drugs, individuals residing in SSA countries are treated in the early stages of the diseases, which has increased their life expectancy to at least 50 years . Most ofUnfortunately, the SSA's ageing research focuses more on health and social sciences, with little to no contribution from humanities, environmental or physical sciences. In most developed countries, environmental scientists  have collaborated with the health authorities to develop accessible neighbourhoods  or housing systems for older adults . This coThis study is the first to provide a systematic mapping review that summarises ageing research in SSA; however, it has some limitations. Because this review is broad, the findings are reported in a Four-Part series; this paper is Part I. The quality assessment and detailed description of specific research areas such as HIV related research and care provision research will be published in subsequent papers. The study's limitations include missed studies due to the number of countries included in the search strategies. In addition, we only included peer-reviewed literature published in English, and some of the national journals in SSA are not indexed in MEDLINE or related databases .We recommend more RCTs that focus on examining the effectiveness of interventions to improve health outcomes among older adults in SSA. National and international funding agencies should set up specific funding competitions to encourage ageing research to develop high-quality RCTs.The SAGE and other longitudinal studies in SSA countries have provided valuable information about risk factors that influence age-related health and wellbeing changes. We recommend that national and international funding agencies develop or commission a longitudinal study to examine life-span development (ageing from childhood to adolescence into adulthood and older age) in the SSA regions. The longitudinal studies could also involve genetic and genomic biomarkers of ageing rather than the commonly used epidemiological variables, including socio-demographic-economic determinants of health.https://erpaan.org/funding.html) are examples of strategies to encourage ageing research in SSA.The growth HIV/AIDs epidemic has levelled off in the SSA since the late 1990s [One reason for the underrepresentation of ageing research from other SSA countries could be a lack of funding to conduct ageing research or lack of funding to publish in an open-peer review. Most open-access journals offer article processing fees (APF) waiver to only researchers residing in lower-income countries. The majority of ageing research was conducted by researchers residing in upper and lower-income countries, eg, Nigeria, South Africa and Kenya. Although open access journals offer at least a 50% APF discount for researchers in middle-income countries, most researchers in this region struggle to pay. We recommend that national and international funding agencies subsidize the APC fees for authors whose articles have undergone peer review and are considered to impact either older peoples' policy or care provision substantially. Committees should be set up to develop strategies, criteria and frameworks to determine ageing articles or studies with a potential of substantial impact in SSA.Transdisciplinary co-production approaches encourage stakeholders' active involvement, including older adults, their relatives and professionals (academic and non-academic) who are not typically part of older adult's care. Such professionals could include engineers who design mobility aids for older adults or adaptive devices for driving assistance among older adults. We did not see such collaborations in ageing research in SSA from the included studies.In conclusion, this review's articles indicated that ageing research in SSA is a predominantly cross-sectional quantitative study design. Noteworthy is the small number of RCTs found in this review. Among the few qualitative studies, descriptive studies represent a higher percentage, while mixed-method studies were the least methodology employed by ageing researchers in SSA. The next series of this review will describe the quality of articles included in this systematic review.


Hepatocellular carcinoma is one of the most common malignancies globally. It not only has a hidden onset but also progresses rapidly. Most HCC patients are already in the advanced stage of cancer when they are diagnosed, and have even lost the opportunity for surgical treatment. As an inflammation-related tumor, the immunosuppressive microenvironment of HCC can promote immune tolerance through a variety of mechanisms. Immunotherapy can activate tumor-specific immune responses, which brings a new hope for the treatment of HCC. At the present time, main immunotherapy strategies of HCC include immune checkpoint inhibitors, tumor vaccines, adoptive cell therapy, and so on. This article reviews the application and research progress of immune checkpoint inhibitors, tumor vaccines, and adoptive cell therapy in the treatment of HCC. Primary liver cancer is one of the common malignant tumors, and its main pathological type is hepatocellular carcinoma (HCC). According to the 2018 cancer statistics of the World Health Organization, the incidence of liver cancer ranks 6th and the mortality rate ranks 4th among the most common cancers in the world (stomach cancer ranks third with a slight advantage) , and theThe liver has a complex immune microenvironment. The liver is continuously exposed to various antigens passing through the portal vein, especially those from intestinal tract. Therefore, the liver microenvironment continues to show immune tolerance, which is to inhibit inappropriate inflammatory reaction and prevent autoimmune liver injury , 9. The The occurrence and progression of HCC are usually accompanied by chronic inflammation  and chronic disease (e.g. liver cirrhosis). Under the action of long-term inflammation, many inhibitory cytokines  are constantly produced, and a large number of immunosuppressive cells, such as regulatory T cells (Tregs), M2 macrophages, and myeloid-derived suppressor cells (MDSCs), are recruited into the liver . FurtherImmunosuppressive cells in tumor tissue can promote HCC tolerance , 13. TumTumor-associated macrophages (TAMs), as one of the key components constituting the immunosuppressive microenvironment of HCC, not only cannot eliminate tumor cells, instead will promote tumor growth , 15.HCC cells can release some cytokines, such as 14-3-3ζ, which can destroy the activation, proliferation and anti-tumor function of tumor-infiltrating T lymphocytes (TILs) . Beyond The expression of immune checkpoints in HCC tissues is increased , 17, 18.CTNNB1  can induce the up-regulation of PD-L1, PD-L2, CD73 and B7-H3; and reversing EMT can inhibit the expression of these markers .In response to these mechanisms, some corresponding measures can be taken to block, inhibit or reverse these mechanisms. For instance, measures can be taken to neutralize inhibitory cytokines or prevent their production. Yang et al.  found thIn recent years, unprecedented progress has been made in tumor immunotherapy. Drugs, therapies and strategies related to tumor immunotherapy are also emerging one after another. Nevertheless, the low response rate and the consequent resistance problem have greatly limited the efficacy of immunotherapy . These mICI is one of the most rapidly developing immunotherapy strategies nowadays. Immune checkpoints are membrane-bound molecules that can be expressed not only on the surface of many tumor cells but also on the surface of numerous immune cells . These iPD-1 is a type I transmembrane glycoprotein expressed on the surface of most immune cells. These immune cells mainly include T cells, NK cells, regulatory cell (Treg), and myeloid-derived suppressor cell (MDSC) , 38. TheIn 2017, the United States Food and Drug Administration (FDA) approved Nivolumab for the treatment of HCC . In one Pembrolizumab is a humanized IgG4/kappa monoclonal antibody against IgG4/K, which can directly inhibit the binding of PD-1 to PD-L1. A phase II clinical trial in 2018 showed that 104 patients with HCC who were intolerant or still progressing after receiving sorafenib treatment were treated with Pembrolizumab . The resFor HCC patients, the high expression of PD-L1 is associated with lower tumor differentiation, a higher level of AFP, more frequent vascular invasion, and worse prognosis . PD-L1 iCTLA-4 is a type of protein receptor located on the surface of T cells. CTLA-4 and CD28 receptors have two ligands in common: CD 80 and CD 86. Compared with CD28, CTLA-4 has a higher affinity with both ligands. CTLA-4-CD80 has the highest affinity, while CD28-CD86 has the lowest affinity , 45. CTLVS (7.2% and 9.6%)] and had an encouraging safety. ICIs can also be combined with other treatment strategies, these strategies include locoregional treatments, antiangiogenetic therapy, chemotherapy, the mammalian target of rapamycin inhibitor, etc. , is the first molecular targeted drug approved for the treatment of advanced HCC. TKI mainly achieves the purpose of anti-tumor by inhibiting the tyrosine kinases of several growth factor receptors. It has been proven that sorafenib can only prolong the survival time of HCC patients by several months. Consequently, it is necessary to develop a combined therapy to further improve the clinical efficacy.Lavatinib, a TKI, has become the first-line treatment for advanced HCC. Finn et al.  combinedVS 98.7%). Currently, Atezolizumab+ Bevacizumab has become the first-line treatment for patients with advanced HCC , microwave ablation (MWA) and cryoablation. Both RFA and WMA deliver large amounts of energy to the tumor site, leading to local heating and destroying tumor cells through thermal efficiency. Cryoablation is to freeze tumor tissue at local low temperature, which induces delayed necrosis of tumor cells after injury.Ablation can also mediate immune regulation . The mecThe principle of tumor vaccines is to introduce tumor antigens into patients in various forms, so as to overcome the immunosuppression caused by tumor, enhance the immunogenicity of tumor cells, activate the immune system of patients, and eventually achieve the purpose of anti-tumor. For the moment, tumor vaccines used for HCC treatment and research mainly include nucleic acid vaccines, peptide vaccines, oncolytic virus vaccines, and DC vaccines.Nucleic acid vaccine refers to the recombination of a gene (DNA or RNA) encoding a certain tumor antigen with a vector, and then injecting it into the patient. After these nucleic acids enter the host cells, the host cells can express the corresponding polypeptides or proteins, thus inducing the body to produce an immune response against these antigens . Once thTumor antigen peptides are important components of peptide vaccines. Tumor antigens must be degraded into short peptides and form peptide-MHC-TCR complex in antigen-presenting cells (APCs) to be recognized by T cells and stimulate the corresponding cytotoxic T lymphocyte (CTL) response. The purpose of peptide vaccines is to deliver a high-dose tumor antigen peptides to major histocompatibility complex (MHC) molecules on the surface of APCs, so as to stimulate the specific immune response of the body to tumor cells (There seem AFP peptide vaccine and GPC3 peptide vaccine which are studied more frequently nowadays. Alpha-fetoprotein (AFP) is one of the most common serum markers in the diagnosis of HCC, and its high expression in hepatocellular carcinoma cells makes it a promising target for vaccine-based therapy . AFP pepOncolytic viruses are a type of viruses that can effectively infect and kill cancer cells. When the virus-infected cancer cells rupture and die, the newly generated virus particles will be released to further infect the surrounding cancer cells. Ovs can not only directly kill tumor cells but also stimulate the immune response of the body and enhance the anti-tumor effect . A generin vivo, and ultimately play the role of anti-tumor (Dendritic cells (DCs) are the most powerful antigen presenting cells (APCs) so far. They are named because they mature with many dendritic or pseudopod-like protrusions. DCs have the function of immune response and immune tolerance, which is of great significance for maintaining immune balance. There exit many inhibitory cytokines in the tumor microenvironment. These inhibitory cytokines can inhibit the normal function of DCs and promote the escape of tumor cells . DC vaccti-tumor , 101.In the phase I and II clinical studies of the DC vaccines in the treatment of patients with advanced HCC, the results showed that the DC vaccines had the tumor-specific immune responses in patients with advanced HCC , 103. Inin situ DC vaccination, that is, direct intratumoral inoculation of unloaded DCs produced in vitro. The unloaded DCs can uptake a variety of TAAs in the tumor, so there is no need to generate TAA vectors in vivo and select specific targets (in situ DC vaccine and immunogenic cell death (ICD) inducer can further improve the antitumor effect of DCs. Because ICD inducers can not only cause tumor cell death and release TAAs but also increase the secretion of damage-associated molecular patterns (DAMPs) that can activate DCs  cells, cytokine-induced killer (CIK) cells and chimeric antigen receptor (CAR)-T cells, etc.ACT is an immunotherapy based on the use of autoimmune cells of cancer patients. The main process is to isolate the immunocompetent cells in the tumor patient, modify these immune cells or stimulate them with some cytokines. These immune cells are amplified and screened NK cells are one of the most important immune cells in human innate immunity, which can produce non-specific immune responses without being sensitized by antigens. The activation state of NK cells is determined by the dynamic balance of the expression of inhibitory receptors and activated receptors on the surface of NK cells. NK cells account for 30% ~ 50% of innate immune cells in the liver , which aNK cells in cancer patients are often in a state of functional failure due to the immunosuppressive effect of the tumor microenvironment (TME). If the failure state of these NK cells can be reversed, it is possible to restore their anti-tumor effect. One animal study  showed tin vitro expansion of CD56Birght NK cells in peripheral monocytes with cytokines. NK cells in peripheral blood can proliferate 140 times in a short time by stimulating with some cytokines . Its mechanism of action is to directly kill tumor cells by releasing perforin and granzyme, or indirectly by releasing a variety of cytokines. Similarly, it can also induce tumor cell apoptosis by activating apoptotic genes. CIK cells mainly include NK-like T lymphocytes (NKT cells) and cytotoxic T lymphocytes. Among them, NKT cells are the main effector cells that exert anti-tumor effects.A retrospective study showed that CIK could significantly improve OS of HCC patients . The resin vitro, and finally injected into patients. TCR-T cell therapy can effectively identify and kill tumor cells by enhancing the specific recognition ability of T cells to tumor cells and improving the affinity of T lymphocytes to tumor cells. Compared with CAR-T cells, TCR-T cells can not only recognize antigens on the surface of tumor cells but also intracellular antigens. Currently, some phase I and II clinical trials exploring TCR-T cell treatment of HCC  are in progress.TCR is a specific receptor on the surface of T cells. It activates T cells by recognizing and binding antigens presented by MHC, and ultimately promotes the differentiation and proliferation of T cells. The principle of TCR-T cell therapy is to introduce TCR genes that specifically recognize tumor antigens into patients’ T cells by gene editing technology, so that these T cells can express corresponding TCR on their surfaces. Then these TCR-T cells are screened and amplified in vitro and combine them with vectors to form recombinant plasmids. Then transfecting these plasmids into patient T cells, which makes these T cells express the corresponding CARs. These CAR-T cells are screened and expanded in vitro, and finally returned to the patient. They can target to kill the corresponding tumor cells after entering the body. Compared with TCR-T cells, CAR-T cells do not require antigen processing and MHC presentation.In recent years, CAR-T cell therapy has become a research hotspot in adoptive cell therapy. CAR is mainly composed of four parts, including, single chain variable fragment (scFv); hinge region; transmembrane region (TM); intracellular signal domain . The principle is to construct CAR genes that recognize tumor antigens Since it was put forward by CAR in 1989, it has developed from the first generation to the fifth generation . The maiThe second-generation CARs add a costimulatory molecule to the first-generation CARs, such as CD28, 4-1BB (CD137), OX40 (CD134) or ICOS, etc. These costimulatory factors can promote the proliferation of CAR-T cells, the secretion of cytokines, and the secretion of anti-apoptotic proteins, thus improving the persistence of CAR-T cells and their ability to kill tumors. Among them, the CAR with CD 28 or 4-1 BB has a stronger tumor-killing effect . FurtherThe third-generation CARs contain two costimulatory molecules, which further enhances the activation, proliferation and persistence of T cells, and makes CAR-T cells have a stronger tumor-killing effect. Moreover, the third-generation CARs that contain both CD 28 and 4-1 BB can provide the strongest anti-tumor effect .The fourth-generation CARs, also known as “TRUCKs”, are modified by adding nuclear factor of activated T cells (NFAT) on the basis of the second or third-generation CARs. For example, the most common NFAT is the cytokine IL-12. These fourth-generation CAR-T cells (IL-12) can release IL-12 after being activated, which can not only promote T cell activation and regulate immunity, but also recruit other innate immune cells to attack tumor cells –131. In The fifth-generation of CARs, are also called “universal CARs”. Theoretically, these CARs can target different tumor antigens. In traditional CAR-T cell therapy, one type of CAR-T cells can only target one kind of tumor surface antigens. In order to improve the flexibility and controllability of CARs and expand the scope of antigen recognition, the fifth-generation CAR adopts a “third-party” intermediate system to separate the antigen binding domain of CAR from its T cell signal unit , 135. Th+ liver cancer cells, and their anti-tumor effect is proportional to the expression level of GPC3 , inhibition of immune checkpoints, etc. With the breakthrough of these obstacles, HCC patients will certainly get more benefits from CAR-T cell therapy.CAR-T cell therapy has made great achievements in the treatment of hematologic malignancies. It also has great potential in the treatment of solid tumors. One of the key factors affecting the curative effect of CAR-T cells is the selection of tumor surface antigens. CAR-T cells are designed for one or more tumor antigens, so they can specifically identify tumor cells expressing these tumor antigens. For CAR-T cells, the best design scheme should be to use tumor-specific antigens (TSAs) to design the corresponding CARs, because these CARs are more targeted and can minimize off-target effects. Unfortunately, there seem too few TSAs, and most of the common targets are tumor-associated antigens (TAAs). Currently, there are some therapeutic targets of CAR-T cells for HCC research and treatment  Lanzhou Talent Innovation and Entrepreneurship Project (2020-RC-38), the Fundamental Research Funds for the Central Universities (lzujbky-2020-kb14), and Major Science and Technology Special Project of Gansu Province (20ZD7FA003).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.