Research on dyslipidemia during pregnancy in women with gestational diabetes mellitus (GDM) has rarely been conducted in Asia. The present study aimed to evaluate maternal mid-trimester lipid profile in relation to GDM and clinical outcomes in these high-risk populations.The medical records of 632 pregnant women in the second trimester were retrospectively analyzed. Maternal fasting serum lipids were assayed for total cholesterol (TC), triglycerides (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), apolipoprotein A1 (Apo A1) and Apo B concentrations during the second trimester. The atherogenic index of plasma (AIP) was calculated as log (TG/HDL). The clinical outcomes were collected by evaluating delivery mode, postpartum hemorrhage, prematurity, macrosomia, birth weight, body length and neonatal Apgar 5 min score.Levels of TG and AIP were elevated while decreased HDL-C was observed in women with GDM compared with that of the control group. Significant differences were observed in gestational weeks at birth, cesarean section, postpartum hemorrhage, birth weight, body length, prematurity and macrosomia between the two groups. Compared with women with hyperlipidemia, the incidence of GDM and cesarean section was lower in normal lipid group. Women in the hyperlipidemia group had smaller gestational weeks at birth than those in the control group. According to the logistic regression analysis, each unit elevation in AIP increased the risk of GDM by 18.48 times . Besides, age  and pre-pregnancy BMI  were the risk factors of GDM.These findings suggested that reasonable lipid control in the second trimester might reduce the incidence of GDM and be a potential strategy for improving clinical outcomes in these high-risk women. Maternal energy metabolism in the second half of pregnancy is directed toward lipolysis, which indicates a physiological adaptation of mothers to maintain stable fuel supplementation to the fetus , 2. MildThe body suffers a substantial increase in the workload of different organs during pregnancy, which may be more likely to result in metabolic diseases. Gestational diabetes mellitus (GDM) is a common metabolic complication in pregnant women and affects up to 22% of pregnancies . PregnanDyslipidemia is a well recognized risk factor for metabolic syndromes. Earlier studies have reported an increase lipid profile in patients with GDM compared to women without GDM . EvidencAlthough changes in lipid profile are expected, there is a shortage of consensus regarding reference values per trimester and the diagnostic criteria for hyperlipidemia. In addition, the use of lipid-lowering drugs during pregnancy is relatively contraindicated. Thus, evaluating the diagnostic criteria of hyperlipidemia and its association with adverse pregnancy outcomes is urgently needed, which may allow for appropriate and timely intervention. In the present study, we examined the blood lipids in patients with GDM, and analyzed putative relations with lipid profiles and clinical outcomes.n = 273 in control group, n = 359 in GDM group) were enrolled in the final analysis. Figure Pregnant women who were admitted to Nanjing Drum Tower Hospital and delivered between January 2015 and December 2017 were recruited for the present study. The inclusion criteria were as follows: (1) age between 20 to 40 years, (2) a lipid measurement at 20–28 gestational weeks, (3) regularly for prenatal examination in the Drum Tower Hospital; We excluded: (1) Incomplete data of medical records; (2) women with other pregnancy complications, medical or surgical diseases ; (3) abortive times ≥3, (4) twin or multiple pregnancy, (5) received assisted reproductive technology, (6) the use of insulin therapy or hypoglycaemic drugs, (7) previous diagnosis of gestational diabetes (or diabetes) and (8) smoking, alcohol abuse and long-term medication treatment (i.e anti-hyperglycemic and anti-hyperlipidemic drugs). Considering a type I error of 5% (α = 0.05), a study power of 80% and the incidence of GDM as key variable, we estimated a sample size of sixty persons for each group at least. Finally, 632 pregnant women , gravidity, parity, permanent residence , monthly income and education attainment. Maternal metabolic parameters included serum TG, TC, HDL-C, LDL-C Apo A1 and Apo B. The atherogenic index of plasma (AIP) = log (TG/HDL). Other clinical data were collected by evaluating delivery mode, postpartum hemorrhage, prematurity, macrosomia, birth weight, body length and neonatal Apgar 5 min score. Preterm birth refers to as birth of newborns less than 37 weeks gestational age. Macrosomia is defined as a fetal birth weight equal to or greater than 4000 g.Pregnant women should subject to GDM screening universally at 24–28 weeks of gestation. And all the women were advised to undergo a regular biochemistry test (serum lipid was included) at the same time. In addition, early glucose screening (<24 weeks) is normally completed at the first prenatal visit in women with risk factors that include obesity with a BMI of ≥30 kg/m2, history of gestational diabetes in a prior pregnancy, known impaired glucose metabolism, hemoglobin A1C of ≥5.6%, first-degree relative with diabetes mellitus, high-risk ethnicity, history of polycystic ovarian syndrome, pre-existing hypertension or cardiovascular disease, or a prior large baby ≥4000 g. Thus, the data were collected at 20–28 gestational weeks.All pregnant women at Nanjing Drum Tower hospital should subject to GDM screening, following an overnight fasting for 8–14 h, universally between 24 and 28 weeks’ gestation. According to the International Association of Diabetes and Pregnancy Study Groups criteria, the diagnosis of GDM was based on a 75 g OGTT . The normal values were fasting plasma glucose < 5.1 mmol/L, 1-h glucose < 10.0 mmol/L and 2-h glucose < 8.5 mmol/L. GDM was diagnosed if one or more values equaled or exceeded the above thresholds .There is no consensus on which criteria should be used in diagnosis of hyperlipidemia. We combined the diagnostic criteria of dyslipidemia in general Chinese population (2016 revised edition) and clinical data, the diagnostic criteria for hyperlipidemia of second and third trimester was established: ①TC > 6.20 mmol/L and/or ②TG > 2.30 mmol/L.p value < 0.05 was considered as statistically significant.To ensure the normal distribution of variables, Histogram and Kolmogrov–Smirnov test were applied. Differences between groups were compared using the independent two sample t test and chi-squared test. Logistic regression analysis was performed to identify risk factors that were significantly correlated with GDM. Confounders were identified based on clinical experience and results of univariate analysis. The relevant confounding variables controlled for the multiple regression analysis included age, pre-pregnancy BMI, parity, gravidity, and gestational age at blood collection, and 95% CIs were calculated. All statistical analyses were performed using SPSS statistical software . A P <  0.001). The pre-pregnancy BMI was significantly higher in the GDM group than that in the control group . There was no significant difference between the two studied groups in terms of gestational weeks, gravidity, parity, residence, monthly income and education attainment , AIP  and lower levels of HDL-C  than that of control. TC, LDL-C, Apo A1 and Apo B were not significantly different between the groups. Regarding the pregnancy outcomes, there was more excessive postpartum hemorrhage  and higher rate of cesarean section  in GDM group than that in control group. The women in control group had larger gestational weeks at birth than the women in the GDM group . As for the conditions of newborn, the birth weight was even lighter in the GDM group . Body length of the newborn in GDM was slightly shorter compared with the control group . In addition, women with GDM had a higher rate of prematurity  while a lower rate of macrosomia . We failed to find significant differences in mean changes of Apgar score (5 min) between the two groups (P > 0.05).Maternal lipid profile in the second trimester can be found in Table P <  0.001) and cesarean section  was significantly increased in the hyperlipidemia group. Compared with women in the normal lipid group, those in the hyperlipidemia group had smaller gestational weeks at birth . No statistically significant difference was seen between the two groups in terms of pre-pregnancy BMI, postpartum hemorrhage, birth weight, the rate of prematurity, macrosomia,and Apgar score (5 min) (P > 0.05).Maternal hyperlipidemia was defined as ①TC > 6.20 mmol/L or ②TG > 2.30 mmol/L . Meanwhile, GDM had a positive correlation with age  and pre-pregnancy BMI . Other lipid variables were almost no contribution to the risk of GDM  was missing. Second, participants recruited in the present study may not represent the general population of pregnant woman due to the selection bias and sample size, which might influence on the analysis of the relationship between mid-trimester lipid profile and GDM. Third, a relatively short duration of observation. Lipid concentrations could change constantly through gestational weeks. Finally, it would be better to perform a longitudinal study to show the association between serum lipids and GDM.In conclusion, this study added to the current knowledge of associations between changes occurred in lipid metabolism during pregnancy and outcomes for mothers and newborns in women with GDM. Further studies are also warranted to establish the standard of hyperlipidemia diagnosis during pregnancy according to local maternal characteristics. More attention should be paid to the mid-trimester lipid profile in all mothers in clinical practice. As for pregnant woman with abnormal blood lipids, close monitoring and lifestyle management during pregnancy should be carried out to prevent pregnancy complications and improve clinical outcomes as much as possible.


Research shows that there are connections among academic burnout, resilience, and life satisfaction in medical students. However, no study has yet examined the temporal relationships between academic burnout, resilience, and life satisfaction among medical students. This longitudinal study aimed to examine the temporal associations between academic burnout, resilience, and life satisfaction and to explore the possible mediating role of resilience in the relationship between academic burnout and life satisfaction among medical students.This is a three-wave longitudinal study covering the preclinical education period of 20 months. From October 2018 to June 2020, a total of 190 students majoring in clinical medicine filled out the Chinese College Student Academic Burnout Inventory (CCSABI), the Connor-Davidson Resilience Scale (CD-RISC) and the Satisfaction With Life Scale (SWLS) three times. Cross-lagged models were constructed to examine the temporal relationships between academic burnout, resilience, and life satisfaction and longitudinal mediation models were constructed to explore the possible mediating role of resilience in the association of academic burnout with life satisfaction.Among medical students, resilience uni-directionally and positively predicted life satisfaction, while academic burnout uni-directionally and negatively predicted life satisfaction. However, the temporal association between resilience and academic burnout was negative and somewhat bidirectional. Resilience had a significant mediating effect on the relationship between academic burnout and life satisfaction in medical students.Medical educators need to identify and take effective measures to combat academic burnout problems which can lead to reduced life satisfaction among medical students. Resilience-based interventions may be promising in buffering the negative impacts of academic burnout and improving life satisfaction. It is recommended that effective resilience-promotion interventions be developed and implemented in medical education to help enhance medical students’ psychological well-being.The online version contains supplementary material available at 10.1186/s12909-022-03326-6. Medical education is a demanding process full of difficulties and challenges, and different stressors medical students encounter in the process of learning medicine may lead to mental health problems such as academic burnout –3. The cAcademic burnout, also known as school burnout or learning burnout, is a kind of burnout syndrome students acquire in the academic environment. Lian et al. defined academic burnout as a state of prolonged negative emotions towards study, inappropriate learning behaviors and reduced sense of academic accomplishment . StudentAn important psychological resource researchers paid much attention to in medical education is resilience. Connor and Davidson defined resilience as a personal quality which enables individuals to thrive when facing adversities . IndividResearch shows that academic burnout among medical students exerted a considerable impact on their psychological health , 23. A tThe protective role of resilience for medical students’ mental health has been demonstrated in a large number of studies , 28–38. Finally, in view of the above-mentioned studies on the close associations between academic burnout, resilience and life satisfaction among medical students, and since previous research has revealed that resilience is an important psychological resource and an essential mental health protector, we hypothesized that resilience may play a mediating role in the relationship between academic burnout and life satisfaction among medical students. Thus, the fourth aim of the present study was to test this hypothesis by using longitudinal mediation models.To summarize, the current research aimed to examine the temporal relationships (i) between resilience and academic burnout; (ii) between academic burnout and life satisfaction; (iii) between resilience and life satisfaction and (iv) to explore the possible mediating role of resilience in the relationship between academic burnout and life satisfaction among medical students by adopting a three-wave longitudinal design.Mage = 18.09 years, SDage = 0.66 years, T1). In October 2019, in the recruitment for subjects for Wave 2, 222 medical students agreed to participate . As for Wave 3, 216 medical students participated in June 2020 . Among the subjects, 190 medical students  completed all three waves of the survey and we used the data from this group of 190 medical students for the cross-lagged analyses in our study  in Wave 1 . Medical students who agreed to participate completed the questionnaires and the informed consent forms online.1), 0.907 (T2) and 0.890 (T3), respectively.Medical students’ academic burnout levels were measured by the Chinese College Student Academic Burnout Inventory (CCSABI) . With re1), 0.947 (T2) and 0.966 (T3), respectively.Medical students’ resilience levels were assessed by the 25-item Connor-Davidson Resilience Scale (CD-RISC) . Student1), 0.913 (T2) and 0.909 (T3), respectively.The 5-item Satisfaction With Life Scale (SWLS) developed by Diener et al.  was used2/df), Goodness of Fit Index (GFI), Adjusted Goodness of Fit Index (AGFI), Comparative Fit Index (CFI), Root Mean Square Error of Approximation (RMSEA) and Standardized Root Mean Square Residual (SRMR). According to Hu and Bentler [2/df < 3, GFI > 0.90, AGFI > 0.90, CFI > 0.90, RMSEA< 0.08, and SRMR < 0.08, the model was of acceptable fit. Mediation analysis was conducted using the PROCESS Procedure for SPSS (2.16.2). All statistical analyses were performed with the significance level set at p < 0.05 (two-tailed).Means, standard deviations and Pearson correlation coefficients of academic burnout, resilience and life satisfaction were calculated using SPSS version 22.0. Three cross-lagged path models were constructed and examined by AMOS version 24.0. For model fit, six indices were adopted, comprising Chi-square divided by degrees of freedom  and in the expected directions. Specifically, academic burnout was significantly negatively associated with both resilience (r range: −.631 ~ −.451) and life satisfaction (r range: −.542 ~ −.350), while resilience was significantly positively related to life satisfaction (r range: .393 ~ .662) in medical students.Table 2 /df = 2.214, p = 0.084, GFI = 0.989, AGFI = 0.920, CFI = 0.994, RMSEA = 0.080, SRMR = 0.032. As can be seen from the figure, the temporal relationship between academic burnout and resilience among medical students seemed reciprocal, with Academic Burnout (T1) negatively predicting Resilience (T2)  and Resilience (T1) negatively predicting Academic Burnout (T2) . However, between time point 2 and time point 3, while Resilience (T2) still negatively predicted Academic Burnout (T3) , the path coefficient from Academic Burnout (T2) to Resilience (T3) was non-significant.Figure 2 /df = 0.981, p = 0.416, GFI = 0.993, AGFI = 0.964, CFI = 1.000, RMSEA = 0.000, SRMR = 0.024. It is clear to see from the figure that academic burnout significantly negatively predicted life satisfaction in medical students, but not the other way around. The standardized path coefficient from Academic Burnout (T1) to Life Satisfaction (T2) was significant  and the standardized path coefficient from Academic Burnout (T2) to Life Satisfaction (T3) was also significant , whereas the path coefficients from Life Satisfaction (T1) to Academic Burnout (T2) and from Life Satisfaction (T2) to Academic Burnout (T3) were non-significant.The three-wave cross-lagged model exploring the temporal relationship between academic burnout and life satisfaction among medical students was shown in Fig. 2 /df = 1.315, p = 0.262, GFI = 0.991, AGFI = 0.953, CFI = 0.997, RMSEA = 0.041, SRMR = 0.034. As shown in the figure, resilience significantly positively predicted life satisfaction in medical students, but not the other way around. The standardized path coefficient from Resilience (T1) to Life Satisfaction (T2) was significant  and the standardized path coefficient from Resilience (T2) to Life Satisfaction (T3) was also significant , while the path coefficients from Life Satisfaction (T1) to Resilience (T2) and from Life Satisfaction (T2) to Resilience (T3) were non-significant.Figure 1 on Resilience at T2 was significant ; the effect of Resilience at T2 on Life Satisfaction at T2 was significant ; the total effect of Academic Burnout at T1 on Life Satisfaction at T2 was significant . According to Hayes [1 on Life Satisfaction at T2 became non-significant after adding the mediator of Resilience  and the indirect effect of Resilience at T2 was significant  with 95% confidence interval excluding 0 , resilience played a full mediating role in the association of academic burnout with life satisfaction among medical students between T1 and T2 [In order to explore the mediating role of resilience in the relationship between academic burnout and life satisfaction among medical students, two longitudinal mediation models were constructed and tested with 5000 bootstrap samples by using Model 4 in the PROCESS Procedure for SPSS (2014). Figure to Hayes , the pre1 and T2 .Fig. 4Lo2 on Resilience at T3 was significant ; the effect of Resilience at T3 on Life Satisfaction at T3 was significant ; the total effect of Academic Burnout at T2 on Life Satisfaction at T3 was significant , so the prerequisite for testifying the mediation model was met [2 on Life Satisfaction at T3 was still significant after adding the mediator of Resilience  and the indirect effect of Resilience at T3 was significant  with 95% confidence interval excluding 0 , resilience played a partial mediating role in the association of academic burnout with life satisfaction among medical students between T2 and T3 [In the second longitudinal mediation model between time point 2 and time point 3, the effect of Academic Burnout at T was met . Since t2 and T3 .We have also tested the differences in academic burnout, resilience and life satisfaction regarding gender for each of the three waves, but results show that there were no significant differences in these three psychological variables in terms of gender for the three waves , an essential psychological resource (resilience) and a key subjective well-being indicator (life satisfaction) among medical students.The result of the first three-wave cross-lagged model shows that the temporal relationship between resilience and academic burnout among medical students was somewhat reciprocal. In our study, between Time 1 and Time 2, academic burnout was found to negatively predict resilience and resilience was also found to negatively predict academic burnout. However, between Time 2 and Time 3, resilience was found to negatively predict academic burnout, but not the other way around. The possible reason for this phenomenon is that we conducted our survey at Time 3 during the COVID-19 online teaching period. Literature shows that medical students were facing great challenges when trying to adapt to the new mode of online teaching , 46. TheThe result of the second three-wave cross-lagged model shows that the temporal relationship between academic burnout and life satisfaction was unidirectional, i.e. academic burnout was found to be a negative predictor of life satisfaction among medical students, but not the other way around. This finding was in line with the conclusion of the seven-year longitudinal study conducted by Hakanen and Schaufeli , which fThe third three-wave cross-lagged model reveals that the temporal relationship between resilience and life satisfaction was unidirectional, i.e. resilience was found to be a positive predictor of life satisfaction among medical students, but not the other way around. This finding was consistent with the result of a six-year longitudinal study by Shek and Liang , which fOur study shows that resilience, as an important psychological resource, played a significant mediating role in the relationship between academic burnout and life satisfaction among medical students. Lower levels of academic burnout were associated with higher levels of resilience, which in turn contributed to higher life satisfaction levels and higher levels of academic burnout were associated with lower levels of resilience, which in turn led to lower life satisfaction levels. Resilience is a personal quality essential for one to cope with adversity and an individual with a high level of resilience can adapt to and overcome adversity despite under continuous stress , 50. MedThe findings of our study have important implications for medical education. First, academic burnout was found to negatively predict life satisfaction in medical students. This finding indicates that medical institutions and educators need to identify effective measures to reduce medical students’ academic burnout levels in order to enhance their life satisfaction levels. Literature shows that academic burnout can be caused by both external factors such as heavy academic workload and a stressful learning environment , and intFirst, self-reported questionnaires were distributed as our data collection method, so response bias and social desirability bias cannot be avoided. Second, the sample of medical students in the current study came from only one medical university in China, so generalizations about conclusions need to be made with caution. Third, in our study, the temporal associations of only one personal psychological resource of resilience with academic burnout and life satisfaction among medical students were explored, but previous studies show that psychological resources such as self-efficacy and self-esteem are also correlated with academic burnout and life satisfaction, which can be examined in future studies. Fourth, the present longitudinal study only covers the preclinical education over a period of 20 months, longitudinal research spanning a longer period of time covering both preclinical education and clinical education is recommended.This study adopted a three-wave longitudinal design to examine the temporal relationships between academic burnout, resilience and life satisfaction, and to explore the possible mediating role of resilience in the association of academic burnout with life satisfaction among medical students. Results show that academic burnout uni-directionally and negatively predicted life satisfaction, while resilience uni-directionally and positively predicted life satisfaction. However, the temporal relationship between academic burnout and resilience among medical students was somewhat bidirectional. Resilience was found to have a significant mediating effect on the relationship between academic burnout and life satisfaction. It is suggested that medical educators pay attention to medical students’ academic burnout problems and they may consider implementing resilience-promotion interventions in medical education to buffer the negative effect of academic burnout and improve medical students’ life satisfaction levels.Additional file 1. Results of the homogeneity tests of 190 medical students as the representative for 212, 222 and 216 subjects respectively in the three waves.Additional file 2. Differences in medical students' academic burnout, resilience and life satisfaction regarding gender for each wave. Additional file 3. Changes in medical students' academic burnout, resilience and life satisfaction regarding gender over the three waves.


Diabetic cardiomyopathy (DCM), a cardiovascular complication of patients with diabetes, is a special cardiomyopathy that is independent of coronary heart disease, hypertension, and valvular disease. Citronellal (CT) is a monoterpene compound generated by the secondary metabolism of plants. In this work, the therapeutic effect and mechanism of CT in DCM were investigated. Experimental diabetic rat models were constructed through a high-fat and high-carbohydrate diet combined with low-dosage streptozotocin (STZ) treatment. CT was intragastrically administered at the dosage of 150 mg/kg/day. The cardiac functions of the rats were evaluated via cardiac Doppler ultrasound. Changes in myocardial structure were analyzed through histopathology. Changes in the representative indices of oxidative stress, namely, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected on the basis of a biochemical test. Related protein levels were assayed via immunofluorescence and Western blot analyses. The DCM rats in the nontreatment group experienced diastolic and systolic dysfunctions, associated with myocardial hypertrophy, fibrosis, and cardiomyocyte apoptosis. Moreover, this condition was concurrent with metabolic disorders, the degradation of SOD activity in myocardial tissues, the increase in MDA content, the abnormal activation of sodium–hydrogen exchanger 1 (NHE1), and the aggravation of cell apoptosis . Myocardial hypertrophy, fibrosis, oxidative stress, and cell apoptosis were obviously inhibited after treatment with CT (150 mg/kg/day). The abnormal activation of NHE1 was recovered under the action of CT. Our study results showed that CT might play a protective role in the treatment of DCM by repressing the abnormal activation of NHE1. Epidemiological studies show that three quarters of deaths among patients with diabetes mellitus (DM) are caused by cardiovascular diseases (CVDs) , 2. DiabCymbopogon citratus Stapf (DC), commonly known as lemon grass, is a perennial herb that is extensively distributed in tropical and subtropical areas. It has been applied in all kinds of foods, chemical engineering, and traditional medical treatment for a long time [ong time . In receong time . Essentiong time , 11, 12.ong time , anticonong time , and gluong time  activitiong time –18. Our ong time .On the basis of the results of the above studies, citronella essential oil was selected as the drug for the prevention and treatment of DCM. This experiment aims to evaluate the therapeutic effect of CT on DCM, discuss its therapeutic mechanism, and further facilitate the research and comprehensive development of the pharmaceutical values of citronella and promote the discovery of safe and effective natural drugs for the prevention and treatment of DCM.CT was provided by Sigma with a purity of ≥95.0%. Streptozotocin (STZ) was purchased from Sigma  and used to induce experimental diabetes. Primary antibodies against sodium–hydrogen exchanger 1 (NHE1), Bax, and Bcl-2 were obtained from Affinity . Superoxide dismutase (SOD) and malondialdehyde (MDA) detection kits were provided by Jiancheng Bioengineering Institute .Male SD rats (180–220 g) were purchased from Henan Experimental Animal Centre and reared in the SPF-level animal house of Xinxiang Medical University at the temperature of 21°C ± 1°C and humidity of 40%–60%. The light/dark period was 12 h, and the rats were fed with pure water and conventional feed. The animal experimental scheme passed the review of the Animal Nursing and Use Commission of Xinxiang Medical University. All experiments were carried out in strict accordance with the suggestions specified in The Guide for Care and Use of Laboratory Animals of the National Institutes of Health.After adaptive feeding for 1 week, the rats were randomly divided into the control group and experimental group. The control group was kept on a normal diet  during the experimental process. In reference to related literature , type-2 The experimental groups were as follows: (1) control group; (2) CT (150 mg/kg/day) group; (3) DCM group; (4) DCM + CT (150 mg/kg/day) group. The corresponding dosage of CT was given to the CT control group and drug treatment group through intragastric administration once per day for 16 consecutive weeks. The same volume of physiological saline was given to the control group and DCM group.After mildly anesthetizing the animals with pentobarbital, the same doctor evaluated cardiac function and structural abnormality by using a VINNO70 color Doppler ultrasonic cardiogram system. Then, transthoracic 2D m-type ultrasonic cardiograms were acquired. The exported ultrasonic cardiogram parameters included left ventricular end-diastolic volume (LVEDv), left ventricular end-systolic volume (LVESv), fractional shortening rate (FS), ejection fraction (EF), stroke volume (SV), and interventricular septal depth (IVSd).Cardiac tissue specimens were weighed and SOD activity and MDA content in myocardial tissues were detected via spectrophotometry–colorimetry in accordance with the kits' instruction manuals (Nanjing Jiancheng).μm slices. The slices were stained with hematoxylin and eosin (H&E) to observe the change in myocardial structure. Meanwhile, 10 cardiomyocytes were selected from each slice under 400× magnification. The short-axis diameters of the cardiomyocytes were measured, and the mean value was calculated to analyze cardiomyocyte hypertrophy quantitatively.Specimens were collected from the atrioventricular ring parallel to the heart, fixed with 4% paraformaldehyde, embedded in paraffin, and cut into 5 The PAS staining method was used to detect the change in the glycogen content of the cardiomyocytes. Five high-power fields (400×) were randomly selected from each specimen. The Image-Pro Plus 6.0 image analysis system was used to determine myocardial glycogen volume fraction .The Masson trichrome staining method and Gomori silver staining method were used to detect the changes in myocardial interstitial and reticular fibers to analyze the cellular and molecular mechanisms of the development of cardiac hypertrophy. The area of myocardial fibrosis was quantified by using a colored high-definition pathological image-text analysis system . The myocardial fibrosis degree was the ratio of the fibrosis area to the whole myocardial area.Tissue slices and NHE1  were incubated overnight at 4°C. The assay was performed by using an IHC detection kit  in accordance with the kit's instruction manual. Positive immunoreaction was visualized by using 3,3′-diaminobenzidine (Beijing Solarbio). Images were captured with a scanning microscope . NHE1 staining was quantitatively analyzed by using Image-Pro Plus 6 analysis software.After hydration, the sections were incubated together with specific primary antibodies overnight at 4°C and then with fluorescent secondary antibodies at 37°C for 1 h. Positive immunoreaction was visualized with fluorescein FITC and CY3 and observed under a fluorescent scanning microscope .Western blot analysis was performed in accordance with the related reference . The prop < 0.05 was considered statistically significant.Statistical analysis was performed with SPSS20.0 software. The data are presented as means ± SD. Before this analysis, the data were subjected to the normality test and homogeneity test of variance. Intergroup comparison was carried out by using one-way analysis of variance, and p < 0.05). However, blood glucose and FINs levels in the DCM group were significantly higher than those in the control group (p < 0.05). The heart weight/body weight (HW/BW) ratio of the DCM group was higher than that of the control group (p < 0.05). In comparison with that of the DCM group, the body weight of the CT treatment group was remarkably increased, whereas the HW/BW ratio was significantly reduced (p < 0.05). Moreover, CT could reduce the blood glucose and FINs levels of DCM rats to a great extent (p < 0.05).The metabolic characteristics of the experimental animals are shown in p < 0.05), and reduced EF, FS, and SV (p < 0.05). After the intragastric administration of CT, DCM rats showed obviously elevated EF, FS, and SV, but reduced levels of LVEDv, LVESv, and IVSd. However, CT showed no influence on normal SD rats . CT treatment could obviously repress DCM-induced cardiomyocyte hypertrophy (p < 0.05) (p < 0.05). This effect could be reversed by CT (p < 0.05) (p < 0.05)  . PAS sta < 0.05) , glycoge < 0.05) . Masson  < 0.05) . These r Figures .The abnormal activation of NHE1 protein facilitates cell apoptosis, and myocardial cell apoptosis is considered as an important factor promoting the genesis and development of DCM. Given this situation, the expression levels of NHE1, the antiapoptotic protein Bcl-2, and the proapoptotic protein Bax in myocardial tissues were detected through the IHC technique and Western blot analysis. As shown in Citronella is a nontoxic and nonmutagenic high-safety medicinal plant when used at conventional dosage, which has a potential therapeutic effect on a variety of diseases , 23. ItsIn patients with diabetes, DCM is a common serious cardiovascular complication, the primary factor causing heart failure and the main reason for deaths in advanced stages. Its pathogenesis mechanism may be related to insulin resistance, hyperglycaemic injury, oxidative stress, and cardiomyocyte apoptosis. In this work, the DCM model was established through treatment with a high-fat and high-carbohydrate diet combined with intraperitoneal STZ injection. In this approach, the high-fat and high-carbohydrate diet would first induce insulin resistance. Subsequently, the injection of low-dosage STZ resulted in islet cell dysfunction and insulin secretion reduction. Reliable type-2 diabetes animal models were thus obtained. Consistent with those in previous reports , the untC. citratus aqueous extract (125–500 mg/kg) to normal Wistar rats orally once per day. After 42 days, the rats presented dosage-dependent weight loss and reduced blood glucose and blood fat [Previous studies have shown that the occurrence of cardiovascular complications in patients with DM can be postponed by reductions in blood fat and blood glucose levels , 28. Adelood fat . Our prelood fat  and thisCT could exert a therapeutic effect on DCM by repressing oxidative stress. SOD activity could be enhanced, and MDA content could be reduced by the oral administration of CT. Previous reports  have ind+ with intracellular H+ at the proportion of 1 : 1 to maintain the intracellular pH value and cell volume under physiological conditions. Moreover, it is the subtype that is mainly expressed in the cardiomyocytes of human beings and mammals [How CT decelerates the development of DCM remains a question. As far as we know, this effect may be correlated with the inhibition of the abnormal activation of NHE1 by CT. NHE1 belongs to the NHE family. As a transmembrane protein that extensively exists in cells, NHE1 mediates the exchange of extracellular Na mammals . Past st mammals , 32. Jan mammals  experime mammals , 35. Ourp < 0.05) and Bax expression levels were significantly higher (p < 0.05) in the DCM rats, indicating that cardiomyocyte apoptosis was enhanced during DCM. After CT treatment, Bcl-2 levels were significantly elevated and Bax levels were apparently reduced (p 0.05) in the treatment group, indicating that CT could repress cardiomyocyte apoptosis and exert a protective effect on cardiomyocytes. This result was identical to the experimental result obtained by Javadov et al. [The increase in cell apoptosis is the main pathological change in the development of DCM. The loss of cardiomyocytes damages cardiac structure and functions. The levels of Na ions are increased due to overactivation of NHE1. Given the exchange between Na and Ca ions, intracellular Ca ion levels also increase, and Ca overload can activate the mitochondrial permeability transition pore, activate the mitochondrial apoptosis pathway, and lead to cell death . The celv et al.  who applThis study provided support for the new function of CT. Specifically, citronella essential oil exerted potential protective and therapeutic effects on DCM. DC, a natural aromatic plant, remitted metabolic disorders, oxidative stress, myocardial fibrosis, and cell apoptosis in DCM rats, thus improving cardiac functions. This cardiac protective effect was associated with the repression of abnormal NHE1 activation.


Globally Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS) is an ongoing public health issue associated with high morbidity and mortality. Efforts have been made to reduce HIV/AIDS-related morbidity and mortality by delivering antiretroviral therapy. However, the incidence and predictors of mortality in border areas like Metema were not investigated. This study aimed to assess predictors of mortality rate among adult HIV-positive patients on antiretroviral therapy at Metema Hospital.Retrospective follow-up study was employed among ART patients from January 1, 2013, to December 30, 2018. Data were entered in Epi-data 3.1 and exported to STATA 14 for analysis. Kaplan–Meier and Log-Rank test was used to compare survival differences among categories of different variables. In bi-variable analysis p-values < 0.20 were entered into a multivariable analysis. Multivariate Weibull model was used to measure the risk of death and identify the significant predictors of death. Variables that were statistically significant at p-value < 0.05 were concluded as predictors of mortality.2  and viral load > 1000 copy/ml  were found to be a significant predictor for mortality among HIV patients on ART treatment.A total of 542 study participants were included. The overall incidence rate was 6.7 (95% CI: 5.4–8.4) deaths per 100 person-years of observation. Being male , STAGE IV , stage III , TB-coinfection , low hemoglobin , BMI ≤ 15.4 kg/mThe incidence of death was high. Being male, viral load, those with advanced STAGE (III & IV), TB co-infected, low BMI, and low hemoglobin were at a higher risk of mortality. Special attention should be given to male patients and high public interventions needed among HIV patients on ART to reduce the mortality rate.The online version contains supplementary material available at 10.1186/s12981-021-00353-z. Globally Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS) is an ongoing public health issue associated with high morbidity and mortality , 2. An eAccessing antiretroviral therapy (ART) has been increased since 2005, globally an estimated 21.7 million people were accessing antiretroviral therapy (ART) and 940,000 people died from AIDS-related illnesses at the end of 2017 , 5, 6. EInstitutional based retrospective follow-up study was conducted from January 1, 2013, to December 30, 2018, in Metema Hospital, which is located in west Gondar province of the Amhara National Regional State. It is 158 km away from Gondar town, 338 km from Bahir Dar , and 908 km from the country’s capital Addis Ababa.The town has a total population of 119,054  . There a2 = 0.93. Based on this, the calculated sample size by using STATA 14 software is 542. Regarding the sampling technique, a record of study participants have been filtered first from the ART database according to their entry time to follow up, next patients have been filtered using age and eligibility criteria, then we give a unique number for the remaining records and select each record for our study using systematic random sampling.All adult HIV-positive individuals who were on ART at Metema Hospital and who enrolled from January 1, 2013, to December 30, 2018, were included. However, transferred in-patients with incomplete baseline data and unknown initiation date of treatment and status of patients were excluded. The sample size was estimated using Log-Rank and Cox proportional hazard model in STATA version 14. The calculation was done based on the assumption that type I error of 5%, power of 80% and the exposure is body mass index (BMI) the most significant variable . SurvivaThe outcome variable was the time event (death) of patients on ART. The independent variables considered in the study were socio-demographic characteristics such as age, sex, level of education, occupation, marital status, clinical factors like WHO clinical stage, CD4 count, functional status, TB/HIV co-infection, opportunistic infection other than TB, drug adherence, viral load, hemoglobin level, body mass index (BMI) and substance use which is a behavioral factor.In this study was based on records as Good , Fair (85–94% or 4–8 doses missed per month) or Poor (less than 85% or ≥ 9 doses missed per month), health workers used reports by the patient or by directly counting the pills to classify adherence. Those patients who adhere greater than or equal to 95% was considered as good adherence; 85–94% fair adherence and Less than 85% was considered as poor adherence . PatientThe hemoglobin level > 10 gm/dl was categorized as normal hemoglobin and hemoglobin ≤ 10 gm/dl was categorized as low hemoglobin level . In thisTo assure the data quality, high emphasis was given in designing a data collection instrument. The training was given for data collectors and supervisors to have a common understanding of the purpose of research and the techniques of data collection. A standard checklist was used to record information extracted from electronic and patient cards. Before the actual data collection pretest was done on 25 patient charts, those charts were not included in the study. A standard checklist was used for recording information from the database and patient cards. This form was developed using the standardized ART entry and follow-up form employed by the ART clinic. Three experienced ART Nurses who were trained on comprehensive HIV care were involved in data collection. Throughout the data collection process, completeness, consistency, and accuracy were checked by supervisors and the principal investigator daily.The data was entered in Epi-data 3.1 and exported to STATA 14 for analysis. Descriptive statistics including proportions, median, tables, and charts was done to describe the characteristics of study participants. Nonparametric estimations of Kaplan–Meier was done graphically and Log-rank test was used to compare the survival probabilities of categorical variables. After fitting the survival model, the proportional hazard assumption was checked using a graphical and global test (goodness test). The model goodness of fit was checked by using the Cox-Snell residual. Cox proportional and parametric models were compared using Akaike Information Criteria (AIC) or Bayesian Information Criteria (BIC). Weibull model was selected since they have low value of AIC and had a good fitness of the Cox Snell residual. Those covariates with a p-value < 0.2 in the bi-variable analysis were again fitted to the multivariable Weibull regression analysis. Both crud and adjusted hazard ratio with the corresponding 95% Confidence Interval (CI) was calculated to show the strength of association. In multivariable analysis, covariates with a p-value of < 0.05 were considered statistically significant.3, and the baseline WHO staging 2470 (45.6%) of patients were at stage I followed by stage III and stage IV 115 (21.2%). Most of the 413 (76.2%) of the study participants had working functional status and three-fourths of the study subjects had good treatment adherence  study participants were females. The median age of study participants was 32 (IQR = 26–39) years. Nearly two-thirds of the study participants were not educated. Almost half of the 278 51.3%) study participants disclosed their HIV status and nearly half of the 274 (50.5%) study subjects are co-infected with tuberculosis (TB) ) per 100-person year of observation. Over the 72 months follow-up period; 4.9 per 100 adult-months (95% CI 3.7–6.3) died during the first 12 months and 0.7 per 100 adult-months (95% CI 0.4–1.1) died during the second year of follow-up. The cumulative survival probability of patients on ART was 77.44% at the end of the study period  survivor curves versus ln  over different categories of variables. The graph of the two categories of tuberculosis and hemoglobin level hazards does not cross each other, which means that the proportional hazard assumption was satisfied Fig. . An addiThe Cox-Snell residuals  were done to check the goodness of fit for all models. The figure shows that the plot of the Nelson Aalen cumulative hazard function with Cox-Snell follows the 45° line closely. Hence, the output shows Cox–Snell residuals of the Weibull model were satisfied the overall model fitness compared with other models. Among all models, the Weibull model was appropriately fitted data compared with others ) times higher than females. Patients with low hemoglobin patients were 4.14 ) times higher risk of death than those with normal hemoglobin. The hazard of death among patients who had a high viral load was 6.7 ) times higher than that of low viral load. Patients at baseline on ART advanced CLINICAL stage IV had 5.64 ) times increased risk of mortality compared to those presenting with WHO clinical Stage I at the initiation of ART ) per 100-person year of observation, consistent with other studies conducted in different areas of Ethiopia , 27. ThiIn this study, the hazards of death among male patients were 2.4 times higher than females in line with other studies , 35, 36,In this study, those ART clients with higher viral load had an increased hazard of death in agreement with the study conducted in seven teaching Hospitals of Ethiopia . This coSimilarly, patients who had advanced WHO clinical stages (stage III and IV) were a higher hazard of death than those in the WHO clinical stage I consistency with studies conducted in Sub-Saharan Africa , 38–41. Patients who had low hemoglobin levels have an increased hazard of death than those who had normal hemoglobin levels. This finding is comparable with the studies conducted in Sub-Saharan Africa , 42–44. Patients with TB-co infected with increased the hazard of death compared to clients not developing TB. This finding is consistent with studies conducted in rural Uganda  and in E2 was 2.4 times higher than those with a BMI ≥ 18.5 kg/m2. This study is similar to studies conducted in Tanzania [In this study, the hazard of death among patients with BMI ≤ 15.4 kg/mTanzania , CamerooTanzania , Uganda Tanzania , and MalTanzania . PatientTanzania .The potential limitation of this study was because of the retrospective nature of the study which lacks completeness of patient records like substance use, provision of opportunistic infection, prophylaxis status, and hospitalization of patients. We could not ascertain that all recorders of mortality were AIDS-related, so all mortality was considered as HIV/ AIDS-related because of the lack of records on the cause of mortality.2, and being male sex had a higher risk of mortality. Special attention should be given to male patients and high public interventions needed among HIV patients on ART to reduce the mortality rate. Nutritional support and close monitoring of patients in the early period of ART treatment initiation is very vital to improve patient survival.In this study, the incidence of mortality was high. Patients on ART who have Advanced WHO patients (stage III and IV), viral load > 1000 copy /ml, hemoglobin ≤ 10 mg/dl, TB co-infection, BMI ≤ 15.4 kg/mAdditional file 1. AIC and BIC value for model comparison of Cox and parametric model


However, the current knowledge of the molecular mechanism of selenite reduction remains limited. In this study, a selenite-resistant bacterium was isolated and identified as Proteus sp. YS02. Strain YS02 reduced 93.2% of 5.0 mM selenite to selenium nanoparticles (SeNPs) within 24 h, and the produced SeNPs were spherical and localized intracellularly or extracellularly, with an average dimension of 140 ± 43 nm. The morphology and composition of the isolated and purified SeNPs were characterized using dynamic light scattering (DLS), scanning electron microscopy (SEM) with energy-dispersive X-ray (EDX) spectrometry, and Fourier transform infrared (FTIR) spectroscopy. FTIR spectroscopy indicated the presence of proteins, polysaccharides, and lipids on the surface of the isolated SeNPs. Furthermore, the SeNPs showed excellent antimicrobial activity against several Gram-positive and Gram-negative pathogenic bacteria. Comparative transcriptome analysis was performed to elucidate the selenite reduction mechanism and biosynthesis of SeNPs. It is revealed that 197 genes were significantly upregulated, and 276 genes were significantly downregulated under selenite treatment. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that genes associated with ABC transporters, sulfur metabolism, pentose phosphate pathway (PPP), and pyruvate dehydrogenase were significantly enhanced, indicating selenite is reduced by sulfite reductase with PPP and pyruvate dehydrogenase supplying reducing equivalents and energy. This work suggests numerous genes are involved in the response to selenite stress, providing new insights into the molecular mechanisms of selenite bioreduction with the formation of SeNPs.Biotransformation of selenite by microorganisms is an effective detoxification (in cases of dissimilatory reduction, e.g., to Se Among these, selenite is the most toxic form for aquatic life and humans because of its high mobility in aqueous environments and high bioavailability. However, elemental selenium is biologically inert and generally thought to be non-toxic (Selenium (Se) is a metalloid that plays a vital role in maintaining human health . It can on-toxic . Further0 particles formed by Se-oxyanion reducing microorganisms, including Azospirillum thiophilum  was investigated intensively using comparative transcriptomics analysis to clarify the possible mechanism of selenite reduction and biogenesis of SeNPs by the isolate YS02. Furthermore, the antibacterial activity of the SeNPs against Gram-negative and Gram-positive bacteria, such as Escherichia coli and Bacillus subtilis, was also investigated to explore the potential of the SeNPs as key assets in the future of healthcare.In the present study, the bacterial strain YS02 exhibiting an efficient selenite transformation ability was isolated from soil. Selenite reduction assay showed that strain YS02 can transform 93.2% of 5.0 mM selenite to SeNPs within 24 h, serving as an eco-friendly cell factory for the biogenesis of SeNPs. However, the exact molecular mechanism of selenite reduction by this strain is not clear. Therefore, the transcriptome response of YS02 cells exposed to sodium selenite (Na2SeO3) was obtained from Sigma-Aldrich . YEP (yeast extract peptone) broth was provided by Shanghai Gu Duo Biotechnology Co., Ltd. . All other analytical grade reagents used in present study were obtained from Sinopharm Chemical Reagent Co., Ltd.  and Solarbio Science and Technology Co., Ltd. .Sodium selenite  was finally selected considering the most promising growth performance and selenite reduction activity.Soil samples were taken from the seleniferous soil of Shitai county, ChiZhou, Anhui Province, China. The primary screening for microorganisms capable of reducing selenite to elemental Se was performed as described by Genomic DNA of strain YS02 was isolated using a Dzup Genomic DNA Isolation Kit  for the identification of isolate YS02. The 16S rRNA gene fragment amplification and sequencing were conducted as described by 32− reduction efficiency and the amount of Se0 produced by isolate YS02 were performed following a previously established protocol (0) formed, samples were centrifuged at 12,000 g for 20 min. The remaining levels of Se(IV) were determined using ICP-OES . Bacterial cells cultured in YEP medium without Na2SeO3 were set as a control. After incubation overnight, the samples were collected using gentle centrifugation .Isolate YS02 was cultured in YEP medium containing 5.0 mM NaTo use transmission electron microscopy (TEM), the pellets were fixed with glutaraldehyde , placed onto carbon-coated copper grids and observed at 80.0 kV on a transmission electron microscope . For scanning electron microscopy (SEM), the pellets were fixed overnight at 4°C with glutaraldehyde , followed by dehydration with a 30%, 50%, 70%, 80%, 95%, 100% ethanol gradient. Finally, the samples were processed for critical point drying and observed with a Hitachi S4800 SEM .The preparation and purification of SeNPs from bacterial cultures of YS02 were conducted using a previously published protocol of g for 5 min. After centrifugation, the supernatant was collected and filtered through a 0.22 μm filter. Then, SeNPs were harvested by centrifuging the resulting filtered supernatant ; the obtained pellet was washed three times with ddH2O and finally resuspended in ddH2O; the suspension was dried in a vacuum freeze-drying system. A mid-infrared spectrum  of the freeze-dried SeNPs  was recorded in the transmission mode using a Thermo Scientific Nicolet iS20 FTIR spectrometer . The baseline-corrected spectroscopic data were collected and manipulated using the OMNIC software (ver. 8.2.0.387); no automatic smoothening was applied, as the spectrum was of appropriate quality.For FTIR spectroscopic analysis, SeNPs were separated from cells and cell debris in the bacterial culture after incubation with sodium selenite by centrifugation at 6,000 2O to obtain a concentration of 200 mg/ml. Two Gram-positive bacteria including Staphylococcus epidermidis (ATCC-51625), and B. subtilis (ATCC-6633), and two Gram-negative bacteria including Pseudomonas aeruginosa (ATCC-47085), and E. coli (ATCC-8739), were used in this study. Briefly, the four bacteria were cultured in a shaking incubator (180 rpm) at 30°C (P. aeruginosa) or 37°C (other organisms) for 24 h, and then seeded in Petri dishes containing agar media. Then a droplet of 50 μl of the SeNPs suspension or a Kanamycin solution (100 μg/ml) was added onto filter paper disks (ø 6 mm) and left to dry. Finally, the disks with the investigated SeNPs and with the standard antibiotic were placed on the agar plates and incubated at 30°C (P. aeruginosa) or 37°C (other organisms). After 24 h of incubation, the antibacterial activity was evaluated by measuring the size of the inhibition zone.The antibacterial activity of SeNPs produced by strain YS02 was measured determined using the cup diffusion method as described by sequencing of the cDNA library was carried out on a BGISEQ-500 platform . After quality control and removing adaptor sequences/low-quality sequences (P. mirabilis ATCC 29906 (genome assembly ASM16075v1) using HISAT . The raw data produced in the present study have been deposited in the National Center for Biotechnology Information (NCBI) database under the accession number PRJNA737579.Isolate YS02 was cultured under the same culture conditions in YEP medium containing 5.0 mM selenite (selenite treatment) or without selenite (control). After batch culturing at 30°C for 14 h, YS02 cells were harvested, quenched in liquid nitrogen immediately for 5 min, and stored at −80°C until use. The samples from control were defined as CK-1, CK-2, and CK-3, while samples from selenite treatment were set as Se-1, Se-2, and Se-3, respectively. The total RNA extraction, RNA integrity assessment, and cDNA library construction were conducted at Beijing Genomics Institute  according to the methods described by equences , the cle2 ) ≥ 1 or ≤ −1 and False Discovery Rate (FDR) < 0.05 were defined to be regulated differently. GO-Term Finder was used to identify gene ontology (GO) terms, and GO enrichment analysis was performed to evaluate significantly over-represented functional categories using Fisher’s exact test with an FDR threshold of 5%. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was conducted to understand high-level functions and utilities of differentially expressed genes . Differential expression analysis between the two groups (CK vs. selenite treatment) was performed using the DESeq2 Bioconductor package. Gene abundance ratio loges DEGs; .−ΔΔCt method.To verify the reliability of the transcriptome sequencing data, several representative DEGs were selected for quantitative analysis. These genes encode key enzymes involved in the glutathione and sulfur metabolism, pyruvate dehydrogenase system, and pentose phosphate pathway (PPP). Primer sequences used in this study are listed in 2SeO3. BLAST search results indicated that the 16S rRNA gene sequence of strain YS02 exhibited 99% sequence similarity to that of Proteus mirabilis ATCC 29906(T). Furthermore, phylogenetic analysis also showed that strain YS02 belongs to the genus Proteus, with the highest similarity to P. mirabilis  and promote plant growth. Moreover, several members of the genus Proteus, such as P. mirabilis YC801  was depleted during the exponential growth phase (between 9 and 15 h), and then nearly exhausted after 24 h of incubation  both inside and outside of the cells. However, the Se0 nanoparticles could be observed mainly in the extracellular space ; a series of bands at ~3,000–2,800 cm−1 ; weak but typical bands at 1,741 cm−1 (stretching vibrations of the C=O group in ester moieties), 1,451 cm−1 (bending vibrations of –CH3/–CH2– groups) and 1,389 cm−1 (symmetric stretching vibrations of carboxylic –COO− groups in amino acid side chains and/or carboxylated polysaccharides); a characteristic pair of bands between 1,637 and 1,537 cm−1 ; a series of overlapping bands at ~1,200–950 cm−1 featuring polysaccharides and, occasionally, phosphate moieties  and lipids . As was mentioned above, these biomacromolecular components are often found in surface capping layers of SeNPs produced by various microorganisms.Thus, in accordance with the FTIR spectroscopic data, the biomacromolecular composition of the obtained biogenic SeNPs most likely includes proteins  as well as indirectly to the amide I region by its bending (scissoring) H–O–H vibrations which are observed at ~1,640–1,650 cm−1  and Gram-positive (B. subtilis and S. epidermidis) bacteria was evaluated by the plate antibacterial test.So far, a lot of metallic and metalloid nanoparticles have been reported to possess antimicrobial activity and thus can be exploited for alternative medicine therapy against multidrug-resistant pathogenic microbes. To evaluate potential biomedical utilizations of the SeNPs produced by YS02, their antibacterial activity against different specific strains including both Gram-negative (P. aeruginosa and E. coli) was more influenced by the SeNPs compared to that of the Gram-positive bacteria as indicated by corresponding inhibition zones. Application of antibiotic (Kanamycin) provided the highest growth inhibition zones with sizes of 18.4 mm and 22.9 mm for E. coli and P. aeruginosa, respectively. However, the SeNPs also showed significant antibacterial effect with inhibition zone sizes between 13.5 and 12.5 mm for E. coli and P. aeruginosa, respectively. For the Gram-positive bacteria, smaller inhibition zones were observed after the application of SeNPs: the sizes of inhibition zones were 11.9 mm for B. subtilis and 9.8 mm for S. epidermidis. Similarly, it was also found that the growth of E. coli and P. aeruginosa was most influenced by PVA/Chitosan/SeNPs nanocomposite while the effect on B. subtilis was the lowest  were generated and sequenced using a DNBSEQ-T7 sequencer, and about 255 million pair-end clean reads (2 × 150 bp long) were obtained. The number and quality scores of the aligned reads are shown in Although selenite reduction mechanisms and SeNPs synthesis have been studied in several bacterial species, they have not been completely clarified . The gloAfter assembling using Bowtie2, a total of 3,153 transcripts were screened and used for DEGs identification. Following comparison with CK (control sample without Se treatment), a total of 473 DEGs were found to have been expressed during Se treatment, among which 197 genes were significantly upregulated, and 276 genes were significantly downregulated . Based op < 0.05), purine metabolism , and histidine metabolism . The pathways involved in the biosynthesis of antibiotics (ko01130), pyrimidine metabolism (ko00240), and the PPP (ko00030) also had high enrichment scores but were not considered as significant. Furthermore, the enriched KEGG pathways for the downregulated DEGs between the control group and Se treatment group are shown in p < 0.05), oxidative phosphorylation , carbon metabolism , and the citrate cycle  were the most enriched pathways.KEGG pathway enrichment analysis for DEGs has shown that complicated metabolic pathways participated in the response to selenium stress. As shown in cysA, cysW, cysU, and CysP  while comparing Se vs. CK, respectively), suggesting the sulfate transport system permease is responsible for the transmembrane transport of selenite in Proteus sp. YS02. The uptake and transport of selenite in E. coli have also been reported to be controlled by the sulfate transport complex ABC encoded by the cysAWTP operon (cysN (encoding sulfate adenylyltransferase subunit 1), cysD (encoding sulfate adenylyltransferase subunit 2), cysH (encoding phosphoadenosine phosphosulfate reductase), cysJ [encoding sulfite reductase (NADPH) flavoprotein alpha-component], and cysI [encoding sulfite reductase (NADPH) hemoprotein beta-component], were all significantly upregulated . Sulfite reductase (cysJI in this case) was reported to catalyze the reduction of selenite/selenate to Se0 with NAD(P)H serving as an electron donor in bacterial isolates Providencia rettgeri HF16 , deoC , tktA (encoding transketolase), and deoB (encoding phosphopentomutase), were significantly upregulated. Furthermore, the expression of aceE (encoding pyruvate dehydrogenase E1 component) and pdhC (encoding pyruvate dehydrogenase E2 component) were also upregulated in the presence of selenite. The pyruvate dehydrogenase system catalyzes the oxidative decarboxylation of pyruvate with the production of acetyl coenzyme A (acetyl-CoA), NADH, and CO2 (The PPP is one of the major sources of reducing power (NADPH) and metabolic intermediates that are required for biosynthetic processes . It is i and CO2 . Taken tnrd (encoding ribonucleoside-diphosphate reductase), hcp (encoding hydroxylamine reductase), cysG (encoding uroporphyrin-III C-methyltransferase), ndh (encoding NADH dehydrogenase), and hisG (encoding ATP phosphoribosyltransferase) were also upregulated, indicating that these enzymes may be associated with oxidative stress defense and maintenance of redox homeostasis in strain YS02. However, the expression of several expected antioxidant protein-encoding genes, such as gsh (encoding glutathione synthetase), gor (encoding glutathione reductase), gorA (encoding glutathione-disulfide reductase), sodB (encoding superoxide dismutase), trxA (encoding thioredoxin), and trxB (encoding thioredoxin reductase), showed no significant change under selenite treatment. This was similar to the findings of gsh, gor, or trxB.Additionally, the reduction and detoxification of selenite ions by microbes may be accompanied by the production of reactive oxygen species that can damage cell membranes or DNA . It is ncysN，CysA，CysP, CysI, CysJ，metF (encoding methylenetetrahydrofolate reductase), dppB (encoding dipeptide transport system permease), frdC (encoding fumarate reductase subunit C), frdD (encoding fumarate reductase subunit D), and sdhA (encoding succinate dehydrogenase flavoprotein subunit). Overall, the upregulation or downregulation of the tested genes were consistent with the results obtained by transcriptome analysis, which indicated that the latter were reliable.The relative expressions of 10 selected DEGs were determined using RT-qPCR  to validProteus sp. YS02 was proposed based on the above analysis. First, the sulfate transporter system (CysPUWA) is predicted to be involved in the uptake of Se(IV) from the extracellular fluid into the cytoplasm. Meanwhile, the PPP and pyruvate dehydrogenase were also activated under selenite treatment and thus produced NADPH or NADH, providing more reducing power for selenite reduction. The produced NADPH or NADH is then transported to sulfite reductase to favor the subsequent reaction. Finally, the reduction of Se(IV) to Se0 is accomplished by sulfite reductase (CysIJ in this case) with NADPH or NADH serving as the electron donor  can be found in the article/YW and QY: conceptualization. YW: methodology and investigation. YS, YJ, and BM: software. JD and JC: data curation. AT and AK: FTIR spectroscopic analysis and interpretation. SH: writing–original draft preparation, supervision, and project administration. SH and AK: writing–review and editing. YW and SH: funding acquisition. All authors contributed to the article and approved the submitted version.This work was supported by the Anhui Provincial Natural Science Foundation, China (2108085QC88 and 2008085MC60), and the Key projects of Anhui Provincial Department of Education (KJ2021A0881). The work of AT and AK was carried out under research theme no. 121032300311-5 of the Russian Academy of Sciences. The funding organizations had no role in study design, data collection and analysis, decision to publish, or preparation of the 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.


Uric acid has strong antioxidant activity, whereas its oxidative damage is closely related to many diseases. We assessed the association between serum uric acid (SUA) levels and premature rupture of membranes (PROM) in pregnant women with gestational diabetes (GDM) in China.In this cross-sectional study, a total of 456 pregnant women were enrolled. Anthropometric parameters for pregnant women were collected within 12 weeks of gestation. Weight gain during pregnancy was obtained from the patients’ records. GDM was diagnosed according to 75-g oral glucose tolerance tests at the 24-28th week of gestation, and SUA was determined simultaneously. PROM was identified as the natural rupture of foetal membranes before the first stage of labour. Logistic models were fitted to identify the presence of PROM using clinical characteristics with (Model 2) or without serum uric acid (Model 1).P < 0.05). Furthermore, GDM women with PROM had lower levels of SUA compared to those without PROM (P = 0.030). The odds ratio of PROM decreased with increasing SUA levels. The area under the receiver operating characteristic curves for PROM based on Model 2 was larger than that in Model 1 .There were differences in BMI, haemoglobin A1c, fasting blood glucose, 1-h postprandial glucose (PG), 2-h PG, insulin levels, triglycerides,weight gain during pregnancy, the rate of macrosomia, fetus birth weight and PROM between women with and without GDM (all Relatively elevated SUA levels at the 24-28th weeks of gestation were associated with a lower risk of PROM in women with GDM. Therefore, SUA may be a protective factor for PROM in GDM patients. The optimal concentration of uric acid in different diseases and different populations needs to be further studied.The online version contains supplementary material available at 10.1186/s12902-021-00736-3. Premature rupture of the membranes (PROM) happens when foetal membranes rupture during pregnancy at any time before the onset of uterine contractions and occurs in approximately 5 to 10% of pregnancies. PROM that occurs prior to 37 weeks of gestation is referred to as preterm PROM . PROM isGDM affects approximately 6% of pregnant women . After an = 1), 2) diabetes diagnosis before pregnancy (n = 15), 3) polycystic ovary syndrome (n = 15), 4) hypertension (n = 8), 5) liver disease (n = 3), or 6) drugs that may affect glucose metabolism (n = 5). Finally, a total of 152 women with GDM and 304 individuals without GDM were included in this study.In this cross-sectional study, subjects were recruited from the maternity clinic of the Central Hospital of Shanghai Fengxian District from November 2016 to January 2019. The patients were included according to the following criteria: 1) 20 to 40 years old, 2) had a pregnancy examination in our hospital, 3) were prepared to complete a 75-g oral glucose tolerance test (OGTT) screening at 24-28th weeks of gestation and 4) were to give birth in our hospital to ensure the integrity of the data. All subjects underwent a medical history evaluation, and those with the following conditions were excluded: 1) a history of smoking (2) was calculated on the basis of self-reported weight measurements before the pregnancy.Pre-pregnancy BMI  levels were measured using high-performance liquid chromatography , and insulin levels were determined using an insulin detection kit and its reagent .One-step 75 g OGTTs were performed during 24-28th gestational weeks. Fasting and postload blood glucose and insulin levels were measured at 0, 60, and 120 min. GDM was diagnosed according to the 2013 World Health Organization diagnostic criteria: fasting blood glucose ≥5.1 mmol/L,1-h OGTT blood glucose ≥10.0 mmol/L, or 2-h OGTT blood glucose ≥8.5 mmol/L.All participants were inpatients before delivery. Weight gain during pregnancy was collected, and primary adverse outcomes were confirmed by experienced obstetricians. The natural rupture of foetal membranes before the first stage of labour was diagnosed as PROM. All subjects had delivered and finished follow-up by December 2019.P-values < 0.05 were considered statistically significant.Continuous data were tested for normal distribution using the Kolmogorov-Smirnov test. Normally distributed continuous data are presented as the Means ± standard deviation, and a t-test was used for comparison. Nonnormally distributed continuous data are presented as medians (range), and a Mann-Whitney U test was used for comparison. Categorical variables are expressed as frequencies (percentages), and a chi-square test was used for comparison. The blood uric acid level of the total population was divided into three groups according to the tertiles. The association between serum uric acid (SUA) level and PROM was assessed using binary logistic regression, and odds ratios (ORs) and 95% confidence intervals (95% CIs) are presented. The predictive value of different models for PROM was analysed using receiver operating characteristic (ROC) analysis. Statistical analyses were performed using SPSS for Windows version 20.0 . Two-sided P < 0.05). Moreover, there were significant differences in weight gain during pregnancy (P = 0.014), foetal birth weight (P = 0.003), rate of PROM (P = 0.033) and macrosomia (P = 0.004) between the two groups. There was a similar level of systolic blood pressure before delivery in the two groups.Table P = 0.030) and uric acid (P = 0.030) between the two groups. There were no significant differences in uric acid between the GDM and non-GDM groups when PROM status was not considered (P = 0.130)  was related to PROM in the GDM group in multivariate analysis. Furthermore, the uric acid of the whole population was stratified by three quantiles, and the results showed that the incidence of PROM decreased with the increase in blood uric acid tertiles. The corresponding ORs (95% CIs) were 0.13 [0.02–0.95] in the second tertile and 0.07 [0.01–0.93] in the upper tertile in comparison to the lower tertile .Binary logistic regression was conducted to assess the association of PROM with clinical variables with PROM as the dependent variable, as shown in Table P < 0.05. When uric acid was added as a variable, the area under the curve in model 2 reached 0.86 [0.75–0.98], P < 0.001.Furthermore, the predictive value of different models for PROM was analysed using receiver operating characteristic (ROC) analysis. As shown in Fig. The present study indicated that women with PROM had lower levels of SUA than those without PROM, and the prevalence of PROM decreased with the increase in SUA tertiles among the GDM group. Additionally, multiple logistic regression analysis indicated that SUA was independently associated with PROM in the GDM group. Finally, the AUC was larger in the model incorporating SUA combined with general clinical parameters than that only considering general indices.What we already know about PROM focuses on infectious and external factors, such as genital infections by chlamydia and/or gonorrhoea, intrauterine infection and cervical abnormalities before pregnancy . StudiesThe concentration of serum uric acid has two sides in the pathophysiological process of many diseases. Hyperglycaemia accelerates purine nucleotide synthesis, which in turn stimulates nucleotide breakdown and increases the concentration of nucleotide degradation products, including superoxide molecules and uric acid . These sReference values for pregnancy can be extremely important for making clinical decisions. Normal reference values are usually based on blood samples from nonpregnant women. However, uric acid levels are higher in women late in pregnancy than in nonpregnant women . AdditioA general health screening study of individuals revealed that SUA levels are positively associated with antioxidant potential; these results indicate that SUA can prevent oxidative damage . GDM is There was some strength in this study. Blood biochemical tests (including SUA) were collected from all subjects at 24-28th weeks of gestation to exclude the effects of differences in gestational weeks on the results. All the subjects were women because of pregnancy, and none of the subjects consumed alcohol. Consequently, possible SUA-influencing factors, such as sex and alcohol consumption, were excluded. Except for the factors that may affect glucose metabolism, there were no other recruitment restrictions, thus the study population should be a good representation. There are some limitations to this study. This was a single centre study, so the number of subjects was limited. The follow-up period was short; therefore we could make no conclusions on the long-term pregnancy outcomes. The related uric acid metabolism pathways also need further study.In conclusion, changes in uric acid levels at physiological concentrations can have adverse effects on PROM, and maternal age is another important factor affecting only pregnant women with gestational diabetes. The moderate increase in uric acid levels at 24-28th gestation may have a potential protective effect on PROM with GDM. At present, we know that the concentration of uric acid is different in different genders and physiological states, and the influence of uric acid on disease outcomes in different diseases is worthy of further study.Additional file 1: Supplemental Table 1. Multivariate logistic analysis of factors related to PROM at different ages in two groups


Escherichia coli, thus overcoming the loss of intracellular metabolites caused by cell membrane damage. After evaluating the extraction efficiency of several extraction methods, we employed the optimized workflow to profile the metabolome of E. coli exposed to cephalexin. In doing so, we proved the utility of the proposed approach and provided insights into the comprehensive metabolic alterations associated with antibiotic treatment.Metabolomics is a powerful tool that can systematically describe global changes in the metabolome of microbes, thus improving our understanding of the mechanisms of action of antibiotics and facilitating the development of next-generation antibacterial therapies. However, current sample preparation methods are not efficient or reliable for studying the effects of antibiotics on microbes. In the present study, we reported a novel sample preparation approach using cold methanol/ethylene glycol for quenching IMPORTANCE The emergence and global spread of multidrug-resistant bacteria and genes are a global problem. It is critical to understand the interactions between antibiotics and bacteria and find alternative treatments for infections when we are moving closer to a postantibiotic era. It has been demonstrated that the bacterial metabolic environment plays an important role in the modulation of antibiotic susceptibility and efficacy. In the present study, we proposed a novel metabolomic approach for intracellular metabolite profiling of E. coli, which can be used to investigate the metabolite alterations of bacteria caused by antibiotic treatment. Further understanding of antibiotic-induced perturbations of bacterial metabolism would facilitate the discovery of new therapeutic targets and pathways. The emergence and global spread of multidrug-resistant bacteria and genes that reduce antibiotic efficacy pose serious threats to human health . The devSaccharomyces cerevisiae  detection. The proposed method prevented substantial loss of intracellular metabolites during quenching of E. coli and determined that the mid-exponential phase was at an approximate optical density at 600 nm (OD600) ≈ 0.35 . Accordingly, all bacterial cultures were subsequently harvested at OD600 ≈ 0.35. Methanol/water (MW)  (60% MW) and liquid nitrogen (LN) are commonly used quenching solvents for the preparation of microbial metabolomics samples  describes the relationship among ADP, ATP, and AMP in cells, reflecting the energy state of biological systems, and can be used to measure the inactivation of cell metabolism . Most ba. lactis –50. Furt lactis –, reflect lactis – and dive lactis –. Differe lactis –, 52. A hE. coli was similar to that in L. plantarum, resulting in an EC value in the range of 0.771 ± 0.003 ∼ 0.849 ± 0.003 (P < 0.05). In contrast to the other three quenching solvents, quenching with LN resulted in a low EC (0.726 ± 0.003), suggesting that LN is less effective at the deactivation of cellular metabolism. Interestingly, LN possessed the lowest temperature (−196°C) but resulted in the worst quenching efficiency. This may be due to serious damage to the bacterial cell membrane caused by the ultralow temperature in the absence of cryoprotectant, causing the leakage of ATP, ADP, and AMP.EC is maintained at a high level if the metabolism of microorganisms is inactivated rapidly using effective quenching solvents. Quenching with 45% ME, 60% ME, and 60% MW were highly effective at inactivating cell metabolism, as the EC values remained within a physiological range, indicating that these methods can effectively inhibit intracellular enzymes and immediately cease cell metabolism . Quenchi ± 0.003 . The higE. coli after quenching. LN-based quenching resulted in similar intensity of SYTO 9 , to evaluate the cell membrane integrity of f SYTO 9  and PI-sf SYTO 9 . For thef SYTO 9 , few PI-f SYTO 9  with 45%13C6]glucose into the bacterial culture before quenching and compared its concentrations in the samples prepared with and without washing. As shown in Fig. S4 in the supplemental material, the percentage of residual [U-13C6]glucose after different quenching approaches using NaCl washing was significantly lower than that without washing (P < 0.05). Next, we assessed the leakage of representative intracellular metabolites, calculating the percentage of leakage in washing groups or the negative control group versus that in the positive control group . The highest and lowest percentages of leakage of the tested metabolites were observed in the LN and 45% ME quenching groups, respectively. No leakage of adenine and adenosine was detected following quenching by 45% ME. There were no significant differences in the leakage rates of other metabolites, except riboflavin, between the 45% ME quenching group and negative control group (P > 0.05).Without the washing step, extracellular metabolites and culture medium components may still remain on the cell surface after quenching and eventually enter samples, interfering with the analysis of intracellular metabolites. To validate the necessity of the washing step, we spiked  + 0.5[ADP])/([ATP] + [ADP] + [AMP]).Energy charge (EC) was used as an indicator to evaluate the inactivation of cell metabolism. After quenching and extraction, intracellular ADP, ATP, and AMP were quantified using a kit , based on enzyme-linked immunosorbent assay by double antibody sandwich method. The ODTo evaluate cell membrane integrity after quenching, the cells after quenching were stained with fluorescent dyes SYTO 9  and propidium iodide (PI) (Invitrogen). The quenched cell pellets were washed twice with sterile phosphate-buffered saline (PBS)  and resuspended in 2 ml of PBS. Cell suspensions (1 ml) were diluted by adding 20 ml of PBS and incubated at 30°C for 1 h, mixing every 15 min. The incubated cell pellets were washed twice and resuspended in 10 ml of PBS. Equal volumes of SYTO 9 and PI were added to the bacterial suspension at a final concentration of 3 μl/ml, and the mixture was incubated at room temperature in the dark for 15 min. The stained bacterial suspension (5 μl) was pipetted onto a glass slide and covered with a coverslip. The stained, quenched cells were visualized by confocal laser scanning microscopy  with a 543-nm He-Ne laser (red channel) and a 488-nm Ar laser (green channel).13C6]glucose  was added into the bacterial culture and mixed, and bacteria were quenched immediately and washed twice with 0.85% NaCl. The pellets were resuspended in the 0.85% NaCl solution, the remaining [U-13C6]glucose was concentrated using C18 cartridges, and the concentration was determined using UHPLC-TOF/MS. To evaluate the leakage of intracellular metabolites after quenching caused by NaCl washing, the NaCl washing solutions from different quenching approaches were collected and concentrated using C18 cartridges, and the metabolites were analyzed by UHPLC-TOF/MS. The positive control group sample was suspended in 0.85% NaCl and heated in a water bath at 80°C for 20 min. Unquenched bacteria were used as a negative control.To investigate the necessity of washing with NaCl,  < 20%, variable importance in the projection [VIP] >1, fold change ≥ 1.5) to identify the differential metabolites caused by antibiotic perturbation. The metabolites were identified using database searches against HMDB (http://www.hmdb.ca/) and ECMDB (http://www.ecmdb.ca) or by metabolite standards . Multivariate analysis and visualization were performed using EZinfo software (Waters), MetaboAnalyst 5.0 , and R (4.0.5 versions), including PCA and orthogonal partial least-squares discriminant analysis.The raw data were acquired using Analyst Software (AB Sciex) and preprocessed for baseline correction, alignment, and peak picking using Progenesis QI v.2.3 . The detected features were filtered (MTBLS3222.The raw metabolomics data have been deposited in the MetaboLights repository under accession number


We explored whether stem cell therapy was effective for animal models and patients with Crohn’s disease (CD).We searched five online databases. The relative outcomes were analyzed with the aid of GetData Graph Digitizer 2.26 and Stata 16.0 software. The SYRCLE risk of bias tool and the MINORS tool were used to assess study quality.n = 567) and 18 human trials (n = 360). In the animal studies, the disease activity index dramatically decreased in the mesenchymal stem cell (MSC) treatment groups compared to the control group. Rats and mice receiving MSCs exhibited longer colons , lower histopathological scores  and lower myeloperoxidase levels . In clinical trials, stem cell transplantation reduced the CD activity index , the CD endoscopic index of severity  and simplified endoscopy score for CD  and improved the inflammatory bowel disease questionnaire score  compared to control values. CD patients maintained high remission rates for 3–24 months after transplantation.We evaluated 46 studies including 28 animal works (Stem cell transplantation is a valuable supplementary therapy for CD.The online version contains supplementary material available at 10.1186/s13287-021-02533-0. Crohn’s disease (CD), a form of inflammatory bowel disease (IBD), is an immune system-mediated, chronic systemic condition characterized by gastrointestinal inflammation and dysregulation of the mucosal-associated immune system , 2. The In recent years, developments in stem cell (SC) biology and regenerative medicine have revealed that SCs unexpectedly can be used to treat autoimmune diseases. Mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) have been shown to counter rheumatoid arthritis, autoimmune hepatitis, and systemic sclerosis . MSCs exFive databases  were searched from their inception dates to February 2021. The search string added the keywords focus on Crohn’s Disease  and stem cells . We also reviewed secondary references. Two researchers independently screened the titles and abstracts of all retrieved articles.Studies that met all of the following criteria were included: single-arm studies or randomized controlled trials including CD patients or animal studies; studies featuring SC therapy with no restriction imposed on the type of SCs, route of administration, or dose; and the inclusion of human CD clinical parameters  or animal disease activity index (DAI) scores, histopathological scores (HSs), colon lengths, myeloperoxidase (MPO) and cytokine levels. Case reports, repeat studies, reviews, and studies lacking full texts were excluded. If more than one article analyzed the same trial, we included only the latest report.Two researchers independently evaluated article quality and extracted data by screening abstracts and full texts. A third researcher was consulted to resolve any disagreements. For animal studies, all relevant data were recorded in Microsoft Excel including the first author; year; location; mouse sex, strain, and weight; group numbers; modeling methods; modeling duration; type and source of MSCs; how MSCs were administered; times of treatment; and other parameters. For clinical trials, the following data were recorded: first author, year, location, size of the MSC group, size of the control group, male/female ratio, type of SCs given, number of SCs administered, administration route, times of treatment, treatment course and follow-up duration. The SYRCLE risk of bias tool was used to evaluate the quality of animal studies , and theI2 test. An I2 value ≤ 50% indicated homogeneity and a fixed-effect model were employed. An I2 value > 50% indicated heterogeneity, and a random-effect model was used instead. Subgroup analyses were performed to evaluate heterogeneity. We employed STATA ver. 16.0 to create forest plots and facilitate the meta-analysis. We used the Begg and Egger tests in STATA (with the significance level set to P < 0.1) to evaluate publication bias. All tests were two-sided, and P < 0.05 was considered to indicate statistical significance.The DAI was calculated from clinical parameters of inflammation  that reflect CD severity. CD morphological and pathological changes were represented by the colon length and HS. The MPO level reflected the extent of neutrophil infiltration. The standardized mean difference (SMD) with the 95% confidence interval (CI) for each parameter was calculated to reveal changes after stem cell therapy in animals. For data from human studies, the means and standard deviations (SDs) of continuous variables  were subjected to SMD analysis. We used odds ratios (ORs) with 95% CIs to determine “remission rates.” Medians with percentiles were converted to means with SDs. If only figures were presented, two researchers independently used GetData Graph Digitizer ver. 2.26 to extract data and compute the means . Among-sA total of 1002 studies were retrieved, from which 262 duplications were initially removed. A review of the titles led to the removal of 380 irrelevant papers; a further 243 were excluded after reading the abstracts. A total of 117 full-text studies were carefully reviewed, of which 61 were excluded for lack of data, 10 because they were off-topic, and 1 because the full text was unavailable. Finally, 28 animal studies –43 and 1A total of 487 mice and 80 rats were used; 78% of all mice were of the C57BL strain, 19.9% were of the BALB/C strain, and 2.1% were of the NOD.CB17-Prkdcscid/J strain. Of all rats, 20% were of the Wistar strain, 42.5% were of the Sprague–Dawley strain, and 37.5% were of the Lewis strain. All 28 studies used one of two models: either CD groups consumed dextran sodium sulfate (DSS) in water while controls received regular water or colitis was induced by intrarectal administration of trinitrobenzene sulfonic acid (TNBS) in ethanol (controls received ethanol only). Information on study characteristics, study quality, and publication bias is shown in Table n = 118 animals), 9 on day 3 (n = 118), 11 on day 5 (n = 134), 3 on day 7 (n = 124), and 5 on day 9 (n = 64). A random effects model was chosen for analysis, and the Cohen method was used to assess differences in DAI between the treatment and control groups. Subgroup analyses showed that the experimental DAI scores were significantly lower than the control scores on day 1 , day 3 , day 5 , day 7  and day 9  . We used a random-effects model to compare colon lengths between treatment and control groups employing the Cohen method. Colon lengths increased markedly in the experimental groups . As the I2 value was high, we performed subgroup analysis by MSC type. Subgroup heterogeneity was low except in the bone marrow-derived MSC (BM-MSC) group  reported rat colon lengths; we again used a random-effect model to compare colon lengths between the treatment and control groups while employing the Cohen method. Colon lengths in the experimental groups were longer than those in the control groups   . Subgroup analysis by the model used showed that the HS for the DSS group  was lower than that for the TNBS group   imparted better effects than did other MSC types , BM-MSC subgroup (I2 = 76.5%), and UC-MSC subgroup (I2 = 22.6%) , the HS was lower in the experimental groups than in the control groups  and heterogeneity was lacking . The meta-analysis indicated that the MPO level in the treatment groups was significantly lower than that in the control groups  : SMD − 3.75, 95% CI − 5.18 to − 2.33, I2 = 77%, P = 0.000], IL-17  and TNF-α  compared with the control group . Subgroup analysis by stem cell type revealed that HSCs  afforded more stable outcomes than did MSCs   was better than the allogeneic stem cells   decreased more than other groups which was injected stem cells twice or fourth times , 68% , 73% , 54% , 52% , and 43% , respectively . In the same way, the SES-CD score of people who received stem cell treatment reduced compared with the score before treatment  . The subgroup analysis showed that the groups injected HSC  had a higher IBDQ score than the groups received BM-MSC treatment    reported the recurrence data and relapses occurred in 29 patients. There was 10 relapsing within one year after the treatment, and the rest 3, 15 and 1 patients presented clinical and/or endoscopic relapse after 12 months, 53.1 weeks and 15 months after transplant, respectively.A total of 7 studies  mucosal healing, and improved the quality of life in CD patients. SC transplantation should be recommended in clinical practice.After subgroup and sensitivity analyses, heterogeneity among studies remained very high, attributable to the injection method and model used as well as differences in stem cell types. Of 28 animal studies, 14 featured intraperitoneal injections, 7 tail vein injections, and the rest other injections. Two different mouse CD models (DSS and TNBS) were employed; DSS concentrations ranged from 1.25 to 5% (w/v), and the model duration ranged from 3 to 34 days. Finally, SC donor age, health status, whether the cells were frozen, endpoints, and whether the cells came from the same species as the recipient may all impact the therapeutic effect. More high-quality clinical and animal trials are required.We compared the responses of animals with DSS- and TNBS-induced CD to MSC treatment. MSCs were therapeutic in both models, but more so in the DSS model. Similarly, in mice with CD, BM-MSCs had a stronger therapeutic effect than did other MSCs.The safety of SC therapy requires attention. We found that the HSC treatment group was more prone to adverse reactions such as viral infections, which often accompany HSC mobilization and regulation. It is clear that CD patients are at higher risk of infection compared to those who undergo transplantation to treat cancer or other diseases that do not involve the intestinal tract. During SC mobilization, patient immunity is reduced and the risk of infection is higher. Therefore, patients should be carefully nursed during mobilization and reasonable drug levels should be prescribed to reduce the development of adverse reactions. To prevent complications in CD patients with perianal disease, it is advisable to perform drainage, implement strict hygiene measures for contact, and prescribe adequate antibiotic prophylaxis . The imp+ T cells are involved in CD initiation and development, and Th1 or Th2 cells are involved in inflammation [+ helper T cells that secrete effector cytokines such as TNF-α and IFN-γ are abnormally high in the guts of CD patients [However, the pathogenesis of CD is complex and remains poorly understood . The wesammation –70. The patients . Immunompatients . The devpatients .Besides MSCs, cytokines and extracellular vehicles (EVs) which are released by MSC also have the therapeutic effect on CD. Because of the low immunogenic profile, which decreases the potential for cell rejection and graft-versus-host-disease, more and more studies begin to focus on the paracrine action of MSC. Neda Heidari et al.  and Ju-HStem cell therapy is not only useful for refractory luminal CD, but also shows good effect on the complications of CD . About 2Our work had certain limitations. First, most human studies were single-arm trials with few patients and thus of low quality. Second, CD-EIS, SES-CD data , IBDQ scores  and pathological grading were lacked in some studies. Any role for SCs in CD alleviation requires further evaluation.We carefully reviewed whether SC therapy improved CD. Stem cell transplantation reduced gut inflammation and improved the quality of life. However, more high-quality randomized controlled clinical trials and basic research are required.Additional file 1:Table S1: The study quality of animal studies; Figure S1: Publication bias of the outcomes: histopathological score, colon length, MPO activity, CDAI; Figure S2: The sensitivity analysis about DAI scores of the 1st day and the 3rd day after the treatment; Figure S3: The sensitivity analysis about DAI score of the 7th day and the 9th day after the treatment; Figure S4: The sensitivity analysis about colon length in the rat group; Figure S5: The subgroup analysis of histopathological score in the mouse group about modeling methods; Figure S6: The sensitivity analysis of histopathological score in the DSS mouse model group; Figure S7: The sensitivity analysis of histopathological score in the TNBS mouse model and the forest plot about the level of MPO activity of animal studies. Figure S8: The forest plot of IL-6, and IL-17 in the both mRNA and protein levels. Figure S9: The forest plot of IL-10 and TNF-α in the both mRNA and protein levels; Figure S10: The sensitivity analysis of HSC subgroup and subgroup analysis of CDAI score of different stem cell sources. Figure S11: The sensitivity analysis of CDAI scores of different treatment times; Figure S12: The subgroup analysis of IBDQ scores of different stem cell types, the forest plots of CRP and the adverse events happened in the experimental and placebo groups.