In the setting of liver transplantation, prediction of hepatocellular carcinoma (HCC) recurrence persists as a fundamental issue, and few tools are available during the waitlist. Biological features such as alpha-fetoprotein values are strong predictors, but it is necessary to understand how we can use them. In this study, an AFP cut-off value was individuated and also its dynamic increase on the waitlist was defined as a predictor.n = 29/207). The median MELD (model for end-stage liver disease) at LT was 12 (9–16). The median time on the waitlist was 92 (41–170) days. The HCC recurrence rate was 16.4% (n = 34/208). The mean time to recurrence was 3.3 ± 2.8 years. The median AFP levels at transplant were higher in patients with HCC recurrence (p < 0.001). At multivariate analysis, the AFP value at transplant that was greater than 25.5 ng/mL (AUC 0.69) was a strong predictor of HCC recurrence after LT . The HCC cumulative incidence function (CIF) of recurrence at 10 years from LT was significantly higher in patients with AFP > 25.5 ng/mL [34.3% vs. 11.5% (p = 0.001)]. Moreover, an increase in AFP > 20.8%, was significantly associated with HCC recurrence (p = 0.034). Conclusions: In conclusion, in our retrospective study, the AFP level at transplant > 25.5 ng/mL and its increase greater than 20.8% on the waitlist were strong predictors of HCC recurrence after LT in a cohort of patients that were waitlisted within the Milan criteria. However further studies are needed to validate these data.Background and Aim: Hepatocellular carcinoma (HCC) recurrence rates after liver transplantation (LT) range between 8 and 20%. Alpha-fetoprotein (AFP) levels at transplant can predict HCC recurrence, however a defined cut-off value is needed to better stratify patients. The aim of this study was to evaluate the rate of HCC recurrence at our centre and to identify predictors, focusing on AFP. Methods: We retrospectively analysed 236 consecutive patients that were waitlisted for HCC who all met the Milan criteria from January 2001 to December 2017 at our liver transplant centre. A total of twenty-nine patients dropped out while they were waitlisted, and 207 patients were included in the final analysis. All survival analyses included the competing-risk model. Results: The mean age was 56.8 ± 6.8 years. A total of 14% were female ( Hepatocellular carcinoma (HCC) is the fifth most common cancer in men (7.5%) and ninth in women (3.4%). Moreover, it is the second most common cause of death from cancer worldwide, being responsible for nearly 9.1% of the total deaths for cancer in 2012  * 100. The best cut-off of ∆AFP was identified by the ROC curve (Youden index). For all of the tested hypotheses, two-tailed Statistical analysis was performed using Stata Software, release 16 .Baseline characteristics are shown in n = 29/207). Of the patients, 35 had a Child-Pugh class C (16.9%) at LT. Median MELD at LT was 12 (9–16). A total of 14 patients had MELD > 25. The median time on the waitlist was 92 (41–170) days: 67 patients (32.4%) 60 days; 54 patients (26.1%) between 61 and 120 days; 49 patients (23.7%) between 121 and 240 days and 37 patients (17.9%) > 241 days.The mean age was 56.8 ± 6.8 years. A total of 29 patients were female .n = 106/207; 51.2%), followed by HBV infection .The median AFP value at transplant was 9.7 ng/mL (4.1–27.1 ng/mL). The most common cause of liver disease was HCV infection (Imaging nearby the LT showed in 135 patients (65.2%) a single nodule of HCC, 2 nodules in 51 patients (24.6%), and 3 nodules in 21 patients (10.1%). 3 (4.2–15.9 cm3).The median diameter of the nodule value among the larger nodules was 2.4 cm (1.8–3 cm), the median value of diameter addition was 3 cm (2–4 cm). The TTV median value was 8.4 cmn = 178/208) underwent bridge therapy while they were waitlisted: 9 hepatic resections (4.3%), 9 ethanol injection (4.3%); 130 percutaneous radiofrequency ablations (RFA) (62.8%); 91 trans arterial chemoembolization (TACE) (44%).A total of 86% of the patients (n = 178/208) of the recipients were on calcineurin-inhibitors (CNI) as backbone immunosuppressive treatment after LT.Likewise, a total of 86% (The baseline characteristics are shown in The mean age was 58.7 ± 17.6 years. In 180 patients (87%), steatosis was inferior to 30%. The mean ischemic total time was 394.8 ± 116.3 min.A total of five transplant procedures utilized a split liver (7.2%).In 167 cases (80.7%), the total number of nodules at explant was 3; in 40 patients (19.3%) was > 3. The median value of the major nodule diameter was 2.5 cm (2–3.5 cm). The vascular micro-invasion was detected in 29 patients (14%). Total necrosis was found in 38 patients (18.4%), partial necrosis in 129 patients (62.3%) and no tumor necrosis was found in 19 livers (9.2%). The predictors of HCC recurrence after LT.n = 34/207). The mean time to recurrence was 39.3 ± 33.6 months. HCC hepatic recurrence was recorded in 11/34 patients (32.3%), five in the first two years.HCC recurrence rate was 16.4% (In 17/34 (50%) patients, HCC recurred only extrahepatically. In 6 patients (17.6%), the time to recurrence was within the first year, in 8 (23.5%) within the second year, and in 20 (58.8%) in the third year.The results of the multivariate analysis are shown in p = 0.01]. The time of waitlist of <6 months was not confirmed in the multivariate analysis as a predictor. At multivariate analysis, the AFP value at transplant was > 25.5 ng/mL (AUC 0.69), Previous HCC bridge therapies were not predictors of HCC recurrence in our cohort.n = 75). A cut-off of 20.8% increase was identified by the corresponding ROC curve  in p = 0.034). We analysed patients with an AFP increase while waitlisted  was stratified for AFP values.HCC RFS was 55.6% (95% CI 47.3–63.1%) after 10 years from LT.p = 0.001)]. We stratified the analysis according to the AFP value and HCC CIF of recurrence at 10 years from LT was significant higher in patients with an AFP > 25.5 [34.3% (95% CI 21.0–48.0%) vs. 11.5% (6.3–18.4%) . We also analysed the cause-specific mortality: HCC-related mortality was 14%, while all other causes of mortality was 27% at 10 years from LT; LT is widely validated as a treatment for HCC, but HCC recurrence is an important negative predictor of post-transplant survival, affecting around 10–20% of the cases .In this setting, it is paramount to identify both patients who can benefit most, as well as avoiding “organ-futility”. Several studies have focused their interest on finding the ideal model to predict post-LT HCC recurrence.p = 0.006) [Initially, the Milan criteria in 1996 included only the tumor burden and number of nodules at explant . In 2012= 0.006) .Our study shows that RFS at 10 years after LT in patients within the Milano Criteria was 55%, as previously demonstrated in the literature .p = 0.009), AFP value at transplant (p = 0.001), and the cirrhosis etiology (p = 0.008) emerged as possible predictors of HCC recurrence; HCV etiology was not a predictor. Previous studies have demonstrated the association between several other factors with post-LT recurrence.Following univariate analysis, the waitlist length (p = 0.06).The RETREAT  score shp = 0.003), while, as demonstrated in previous studies, the radiological response to downstaging could improve the prognostic score [The recent XXL trial  in a groic score .In the setting of patients that are affected by HCC on the waitlist for LT, the concept of prioritization is often debated. Some authors have suggested a new paradigm in which the response to bridging or downstaging represent the main drivers for patient selection and priority allocation. Both the response or the lack of response to neoadjuvant therapies could positively impact prioritization, always within the eligibility criteria of LT .This approach was also studied by Di Sandro et al., where the authors showed how that were patients at higher risk on the waitlist could beneficiate from an early LT (within two months), while for patients with an intermediate risk it would be better to wait to evaluate tumor behavior .p = 0.34). It is worth noting that one of the strengths of the present single-centre study is represented by the presence of a multidisciplinary team, with stable and coherent selection criteria: it is somewhat expected that bridge procedures on Milan-in HCC waitlisted patients might not significantly influence HCC post-LT recurrence.In our study, the execution of bridge therapy to transplant with loco regional treatment did not impact recurrence rates  inhibitors as the main immunosuppressive agents are associated with a lower incidence of HCC recurrence after LT. The SILVER study showed an improvement in recurrence-free survival in the first three to five years in patients that were treated with sirolimus vs calcineurin inhibitors (CNI) .p = 0.03): However, we must consider that, in most cases, the switch of immunosuppression to mTOR was made after the diagnosis of post-LT HCC recurrence.In our study, following univariate analysis, patients that were in treatment with mTOR had a higher rate of HCC recurrence  [Moreover, it has been shown that extremely elevated AFP levels (>1000 ng/mL) before LT is associated with the worst post-LT survivals, regardless of the tumor burden . However = 0.02) .p = 0.01). Moreover, the subgroup analysis showed that an increase in AFP values > 20.8% on the waitlist correlated with higher HCC recurrence rates (p = 0.03). These findings confirm the pivotal role of AFP as an “alarm bell” of HCC recurrence in Milan-in-HCC LT recipients.In our multivariate analysis only, the AFP values at transplant were confirmed as HCC recurrence predictors: indeed, AFP > 25.5 ng/mL at the time of LT predicted a greater than three-time increased risk of post-LT HCC recurrence . This study highlighted the utility of the dynamic AFP-R at predicting the patient outcomes after liver transplant for HCC .p < 0.001) [Regarding the role of AFP in this setting, Mahmud et al. showed that, in a cohort of 1164 transplanted patients, pre-transplant AFP < 500 ng/mL had a 1.6-fold higher risk of death versus those with AFP < 20 ng/mL (< 0.001) .Moreover, Koch et al. showed that in patients with an AFP ratio (AFP at recurrence divided by AFP 3 months before recurrence) of 0.5, HCC recurrence was greater than 70 months, compared to a median of 8 months in patients with a ratio of 0.5. It underlines that a rise in AFP levels could identify patients that are at short-term risk for HCC recurrence after LT .The AFP threshold that was identified in this study could optimize both patient selection and also improve surveillance protocols after LT. Post-LT surveillance should be intensified during the period of the highest post-LT recurrences; unfortunately very few studies were aimed at this issue, which leaves the frequency and timing of both imaging and serum testing unclear.At present, there is no specific evidence-based risk stratification criteria that can be strongly recommended. In general CT or MRI should be performed every six months for 2–3 years after LT, and AFP every six months for 5 years post LT .Our results confirm the potential role of AFP to reduce HCC recurrence and improve the outcomes after LT; it is fundamental to understand the importance of AFP to avoid transplant “futility” and to make decisions in a multidisciplinary setting.This study presents several limitations. Firstly was the selection of patients that were represented only by patients within the Milan Criteria. This reflects the past policy of the transplant centre; in fact, 21 patients were excluded from the analysis for disease progression. Secondly, the relatively small sample size and also only the two measurements of AFP during the waitlist as well as the retrospective design. On the other hand, a single-centre study should usually warrant homogeneity. We believe that our results could contribute to stratify LT recipients and eventually improve post-LT outcomes.In conclusion, in our retrospective study, an AFP value at transplant >25.5 ng/mL and its dynamic increase that was greater than 20.8% while on the waitlist, were predictors for HCC recurrence after LT in a cohort of patients within the Milano criteria. The AFP cut-off could help to stratify patients but it is its increase on the waitlist that could highlight that patients needed more attention on two sides. It could be a reason to prioritize patients for LT, maybe representing the “favorable window” to avoid future graft futility for disease progression. On the other hand, if the increase continues to persist and patients are always eligible for LT, physicians have to optimize immunosuppression and strictly monitor the recurrence after LT. These data could be used to correctly stratify HCC patients before LT and improve surveillance after LT, but further studies are needed to validate these results.


To compare the middle-term efficacy and safety results between scrotoscope-assisted (SA) minimally invasive excision and traditional open excision (OE) for the treatment of epididymal mass.A total of 253 males with surgery excision of epididymal mass from 2012 to 2018 were included in this retrospective study. Patients were divided into two groups: the traditional OE group and the SA group. Patient demographics and intraoperative and postoperative outcomes were obtained and compared between these two groups.About 174 patients (68.8%) underwent SA, and the other 79 (31.2%) underwent OE. Demographic data were similar between the two groups. Compared with OE surgery, SA could significantly shorten the operating time (19.4 ± 4.1 vs. 53.8 ± 12.9 min), reduce blood loss (5.3 ± 1.5 vs. 21.3 ± 5.6 ml), and downsize the operative incision (1.5 ± 0.3 vs. 4.5 ± 0.8 cm). Additionally, postoperative complications were significantly less occurred in the SA group than those in OE (15.5% vs. 21.5%), in particular scrotal hematoma (1.7% vs. 12.7%) and incision discomfort (2.8% vs. 6.3%). Patients in the SA group had a significantly higher overall satisfaction score (94.8 ± 3.7 vs. 91.7 ± 4.9) and a significantly shorter length of hospital stay (4.1 ± 0.9 vs. 5.0 ± 1.5 days) than those in the OE group. No postoperative testicular atrophy occurred in the SA group.SA is emerging as a novel and effective option with promising perspectives for epididymal mass therapy. Epididymal mass is recognized as a common disorder in the male population but still seems to be a diagnostic and therapeutic dilemma. The most prominent types of epididymal mass are mass-forming epididymitis , 2, epidFor the surgical treatment of epididymal mass, the traditional open excision (OE) of mass is one of the main choices. However, OE is a non-minimal invasive treatment for scrotal diseases and brings relatively more postoperative discomfort and complications  , 12. FirA retrospective analysis was performed at the Second Xiangya Hospital and the Fujian Provincial Hospital from January 2002 to 2018. Study approval was obtained from the Ethics Committee of Fujian Provincial Hospital (K-2019-10-03). Epididymal mass was diagnosed by scrotum ultrasound. The patient was included under the following conditions: aged 18 to 60 years old, diagnosed with epididymal mass, and failed conservative treatments with or without obvious symptoms. All patients underwent a clinical assessment including vital signs, ECG, and laboratory examinations . We conducted C-reactive protein, purified protein derivative and chest radiography, urinary ultrasound or KUB + IVP, CT, and/or scrotum MRI to exclude epididymis tuberculosis or malignant tumor. Cases with severe cardiopulmonary diseases or coagulation disorders were also excluded. All patients were fully informed about the advantages and disadvantages of each surgical approach, and the patients had their own right to determine which one to choose.All patients received general anesthesia and operated in bladder lithotomy position with routine skin preparation. Disposable plastic incise drape was pasted in the operation area after sterilization.The SA procedure was performed as previously described , 17, 20.For those patients treated by the OE procedure, an about 4-cm anterior scrotal incision was made in the ill side. The epididymis and the testicle were taken out of the incision, and the epididymal mass was inspected and excised. Like SA, the incision was then closed, and a drainage strip was also used and removed within 24–48 h.A descriptive analysis of patient demographics was performed, including age, time since onset, follow-up period, and the characteristics of epididymal mass (location and size). The mass size was defined as the greatest diameter recorded on ultrasound recording. Surgical details mainly included intraoperative  and postoperative  results. Postoperative complications were recorded in detail and graded by Clavien–Dindo system, which could be applied as a widely used tool to assess and report postoperative complications in general surgery , 22. Allt-tests and used a p-value of less than 0.05 as a cutoff for statistical significance. A multivariable logistic analysis with a likelihood ratio test was performed to identify predictors of satisfaction. All data entry and analysis were carried out in the SPSS 24.0 statistical analysis software .Data were expressed respectively in terms of mean ± SD for normally distributed continuous values and as median with range for non-normally distributed data, whereas discrete ones were reported using percentages. We compared continuous variables with A total of 253 patients with epididymal masses were enrolled in this retrospective study, 174 underwent SA and 79 underwent OE. The mean age was 47 ± 12.8 (22-80) years in SA and 48 ± 14.9 (22-80) years in OE, respectively. The mean follow-up time was 20.8 ± 8.2 months in SA vs. 19.4 ± 8.6 months in OE. All patients underwent surgery successfully. The mean operating time in SA was significantly shorter than OE (19.4 ± 4.1 vs. 53.8 ± 12.9 min). The blood loss in SA was significantly less than OE (5.3 ± 1.5 vs. 21.3 ± 5.6 ml). The mean incision size in SA was significantly shorter than that in OE (1.5 ± 0.3 vs. 4.5 ± 0.8 cm).p < 0.05).Patients in the SA group had a significantly less frequency of dressing changes (2.9 ± 1.3 vs. 4.4 ± 1.7 times) and a significantly shorter length of postoperative hospital stay (4.1 ± 0.9 vs. 5.0 ± 1.5 days) when compared with the OE group . A signiNo significant difference was reported for the total incidence of complications and complication Grades, which is 27 (15.5%) in the SA group and 17 (21.5%) in the OE group (Grades I–III) . Among tOur study firstly highlighted the feasibility of scrotoscopy in the treatment of epididymal masses. As shown by our results, SA demonstrated significant superiority over traditional OE for the treatment of epididymal mass.Traditional open resection of epididymal mass or open-partial epididymectomy was most commonly chosen for epididymal mass surgical treatment in the patients who failed medical treatment with/without obvious symptoms, or suspected malignant tumor –10, 24. Consistently, we applied scrotoscopy for the treatment of epididymal masses and achieved superior results over the traditional approach. In this study, SA showed significantly less operating time, less blood loss, and a shorter length of incision than that in OE. Possible reasons for these results are given below. Mean operating time in SA was less than OE because more time was needed to achieve hemostasis in OE. The endoscope can easily insert into the cavity of the tunica vaginalis and can easily observe the space-limited cavity, so the incision does not need to be as long as the OE. With the application of electric cutting technology, no obvious bleeding was observed during the SA process. These results support that scrotoscope could serve as a feasible and efficient tool for the treatment of epididymal mass.Re-emphasizing the safety for scrotoscope to treat epididymal mass, the SA group showed fewer postoperative complications and a faster recovery when compared with the OE group. According to the Clavien–Dindo grading system, all the postoperative complications in two groups were Grades I–III. One Grade III case was reported in the OE group, but none in the SA group. Scrotal edema was the most common complication in SA; this may be related to the damage of epididymis or extra fluid infiltration into the interlayer of the scrotal wall through the incision. Generally, avoiding wall sheath damage and reducing perfusion pressure and time can effectively avoid edema , 20. AddHowever, certain limitations should be addressed in this research, including the retrospective nature, the limited size of the total population, and lack of long-term follow-up data. Furthermore, the SA and OE groups were not perfectly matched by age, size of lesion, and etiology for the operation. The estimation of blood loss in the SA group may be inaccurate, which would reduce the confidence power. Moreover, the postoperative length of stay was relatively long for such a scrotal surgery under scrotoscope, possibly due to previous local medical insurance policy. In addition, patients with SA preferred to stay longer due to the concern of this uncommon surgical procedure to the public. Future studies with larger sample sizes should be conducted to further validate its diagnostic and therapeutic value.In conclusion, our present study confirmed that SA was a safe and effective minimally invasive therapeutic option for epididymal mass. It has the advantages of small incision, rapid recovery, and low risk of complications. It is worthy of further clinical application.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by K-2019-10-03/Ethics Committee of Fujian Provincial Hospital. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements. Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.YW and YG conceived of the presented idea and encouraged JY, RZ, YY, WC, TL, QZ, and LY to collect data and supervised the findings of this work. YW developed the theory and performed the computations. CQ verified the analytical methods and wrote the manuscript with support from YG. All authors discussed the results and contributed to the final manuscript.Support was provided by the Startup Fund for scientific research, the Fujian Medical University (Grant number: 2019QH1154), and the Natural Science Foundation of Fujian Province (Grant number: 2021J01359).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.


Plasmodium falciparum was the predominant species followed by Plasmodium malariae and then Plasmodium ovale. The RDT and LM had the same sensitivity (90.6%) with a slight difference in their specificity . The RDT also demonstrated higher positive and negative predictive values compared with those of the LM.Malaria is still a threat to public health as it remains the first endemic disease in the world. It is a pervasive parasitic disease in tropical and subtropical regions where asymptomatic malaria infection among humans serves as a significant reservoir for transmission. A rapid and correct diagnosis is considered to be an important strategy in the control of the disease especially in children, who are the most vulnerable group. This study assessed the prevalence of asymptomatic malaria in children at the Nkolbisson health area in Yaoundé, Cameroon. A cross-sectional study design and a convenience sampling plan were used. A total of 127 participants were recruited after informed and signed consent from parents and/or guardians. Blood samples were collected by finger-pricking and venipuncture from children aged 6 months to 10 years and then screened for asymptomatic parasitemia by a rapid diagnostic test (RDT), light microscopy (LM) staining with Giemsa and 18S rRNA polymerase chain reaction (PCR) for speciation. The data were analyzed using SPSS version 20 software. The study identified 85 children who were positive from the PCR, 95 positive from the RDT and 71 from the LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively. The prevalence was not observed to be dependent on the sex and age group of the participants.  Plasmodium species have been identified as causing malaria with Plasmodium falciparum accounting for the greater proportion of the global malaria burden especially in children under the age of 5 years [Plasmodium falciparum infection can result in a disease manifested by the asymptomatic carriage of uncomplicated or severe malaria [Plasmodium falciparum and non-falciparum species in children resident in Nkolbisson, Yaoundé, Cameroon.Malaria is still a threat to public health as it remains the first endemic disease in the world. It is a pervasive parasitic disease in tropical and subtropical regions. It is mostly prevalent in Sub-Saharan Africa, Asia and Latin America ,2. The S 5 years . Malaria malaria ,7. Asymp malaria . About 2 malaria ,7. In ma malaria . Most as malaria . The stu malaria . Evidenc malaria . Underst malaria . To comp2. Yaoundé is located 700 m (2300 feet) above sea level and at 3′52′ N 11°31′ E. The climate is of a Guinean-type with two rainy seasons (March–June and September–November) and two dry seasons (December–February and July–August). The average annual rain fall here is 1643 mm with a relative humidity ranging from 85 to 90% [The study was carried out in the Nkolbisson health area in the Cite-Vert health district in the Centre region of Cameroon where Yaoundé is the capital city. Yaoundé has a population of about 2,500,000 inhabitants and covers a surface are of 183 km5 to 90% ,14,15. MThe study received ethical approval from the National Ethics Committee for Human Health Research (N° 2018/07/1091/CE/CNERSH/SP). Research authorizations were also obtained from the Centre Regional Delegation for Public Health and from the health centers where the study was conducted. Prior to the participant enrollment, written and signed consent from each parent or guardian was obtained based on the informed consent forms designed for the study. The potential risks and benefits as well data privacy and confidentiality were clearly explained to parents or guardians. Only children whose parents or guardians signed and returned the consent form were included in the study.This study was a cross-sectional study and a convenience sampling approach was used. All children resident in the study area aged 6 months to 10 years were eligible. All participants were examined by a physician for malaria. Children who presented signs and symptoms of malaria such as a headache, tiredness or fever were not enrolled. Finger-pricking for blood samples was performed on children whose parents or guardians consented to the study. A questionnaire was used to collect the demographic data and vital signs as well as to assess the knowledge and attitude of the participants toward malaria prevention and control. A total of 127 participants were recruited between November 2019 and February 2020.Plasmodium species-specific PCR were blindly submitted to a second independent microscopist. Venous blood samples were collected from each patient by a syringe. A rapid diagnostic test (RDT) was performed on the spot in the field using a malaria rapid diagnostic kit supplied by SD Bioine (batch number: 05EDD011B) for the National Malaria Control Program in Cameroon to detect two antigens of the malaria parasites (HRP2/pLDH) that were at the different health centers, as described by the WHO . About 6Genomic deoxyribonucleic acid (DNA) was extracted from 200 mL of blood in EDTA tubes using an EZNA DNA extraction blood kit  according to the manufacturer’s instructions. Prior to the extraction, a few materials were sterilized by autoclaving and the equipment was disinfected with 10% bleach and 70% alcohol. The blood samples were briefly thawed at room temperature. In a sterile Eppendorf tube, 100 µL blood was introduced with the help of a micropipette. A total of 150 µL of elution buffer (65 °C) was then added onto it. An OB protease and a BL buffer (lysis buffer) was then added to the mixture and vortexed for 15 s, after which it was incubated at 65 °C for 10 min. During the incubation, the tubes were briefly vortexed twice. A total of 260 µL of 100% ethanol was then added to the mixture, vortexed for 20 s and then centrifuged briefly. The mixture was then transferred to a DNA spin column (DNA HiBind Column) and centrifuged for 1 min at 10,000 rpm. A total of 700 μL of a wash buffer was then introduced into each column. This step was repeated for each tube. A 2 min centrifugation at 10,000 rpm was performed to dry the column matrix. The columns were transferred into a nuclease-free microcentrifuge tube. A total of 150 µL of elution buffer was used to elute the DNA, which was then stored at −20 °C for a further analysis.Plasmodium species was performed in accordance with a protocol described by Putapomtip et al. [Plasmodium genus-specific outer primers . A volume of 25 µL from the first reaction contained 20.25 µL of nuclease-free water, 2.5 µL of 10 X thermopol buffer (New England Biolabs), 0.5 µL of 10 mM dNTPs, 0.25 µL of primer, 0.25 µL of Taq polymerase (New England Biolabs) and 1 μL of amplicons. This second step took place under the following conditions: 95 °C for 15 min, 30 cycles of 58 °C for 2 min, 72 °C for 5 min and 94 °C for 1 min followed by 58 °C for 2 min and 72 °C for 5 min.A PCR was performed on all samples and the 18S rRNA gene was used to detect and identify the different malaria-causing species. The PCR amplification of the p et al. . A neste primers  and 0.25A total of 127 children were enrolled into the study . There wPlasmodium falciparum malaria infection by the RDT, 85 (66.9%) by light microscopy and 71 (55.9%), by the PCR. From the patients screened by the PCR, 66.9% were positive, among which were 51 (75.0%) males and 34 (57.6%) females. A total of 42 (33.1%) children were negative for malaria by the PCR. Of the 95 (74.8%) children diagnosed positive for Plasmodium falciparum malaria by the RDT, 54 (79.4%) were males and 41 (69.5%) females. In addition, 32 (25.2%) of the children were diagnosed negative for Plasmodium falciparum malaria by the RDT, among which were 18 (30.5%) females and 14 (20.6%) males. When the children were diagnosed for Plasmodium malaria using microscopy, 71 (55.9%) of them were positive and 56 (44.1) negative. Among the positive cases 17 (28.8%) were females and 54 (79.4%) were males whereas among the negative cases, 42 (71.2%) were females and 14 (20.6%) were males (p = 0.199) and microscopy (p = 0.282). However, an association was observed between sex and malaria prevalence by the PCR (p = 0.038). No significant association was observed between the age group and malaria prevalence, be it by the RDT (p = 0.508), microscopy (p = 0.682) or by the PCR (p = 0.655).The three diagnostic methods, light microscopy (LM), a rapid diagnostic test (RDT) and a polymerase chain reaction (PCR), detected significant malaria infections in asymptomatic children. A total of 95 (74.8%) from the 127 cases detected positive for a re males . A high re males . No signPlasmodium falciparum, 4 were infected with Plasmodium ovale and 7 were infected with Plasmodium malariae. Of the 85 infected with Plasmodium falciparum, 51 (75.0%) were males and 34 (57.6%) were females. A statistical significance was observed between the gender of the children and infection by Plasmodium falciparum (p = 0.038)  was female and 3 (4.4%) were males. Furthermore, 4 (5.9%) of the 7 children infected by Plasmodium malariae were males and 3 (5.1%) were females. No statistical significance was observed between the gender in children infected with Plasmodium ovale (p = 0.382) as well as Plasmodium malariae (p = 0.844)  . Of the = 0.844) .Plasmodium falciparum malaria by microscopy and who were truly infected with Plasmodium falciparum was 80.2% and that of those who were diagnosed negative and who were truly negative was 74.2%. These were the respective positive and negative predictive values for microscopy. The positive and negative predictive values of the RDT stood at 81.1% and 75.0%, respectively.Overall, when light microscopy (LM) and RDT diagnostic tools were compared with the PCR as the standard, 77 out of the 85 children that were diagnosed positive for malaria by the PCR were also positive by microscopy, giving a sensitivity or true positive rate of 90.6% with a false negative rate of 9.4% . Of the p = 0.532) or between the age groups of the children (p = 0.698).The average parasitemia of each child was 8260.19 ± 20,427.84 parasites/µL . Among tPlasmodium falciparum and non-falciparum species in children resident in Nkolbisson, Yaoundé, Cameroon. LM, RDT and PCR malaria diagnostic techniques were used to detect Plasmodium falciparum and non-falciparum species in children between the ages of 6 months and 10 years. The study compared the diagnosis of malaria by LM, a RDT and a PCR with the assessment of asymptomatic malaria in children. A total of 127 asymptomatic children were tested for malaria. The study identified 85 children as positive by the PCR, 95 positive by the RDT and 71 by LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively.This study assessed the prevalence of asymptomatic malaria in children in Nkolbisson, Yaoundé. Nkolbisson is in a forest malaria transmission geo-ecological zone where malaria transmission is year-round . The conBased on LM (55.9%) and the RDT (74.8%), the study reported a high prevalence of asymptomatic malaria in children. This prevalence was higher compared with that observed in other studies carried out in Cameroon; for instance, in Buea where a prevalence of 40.32% from LM and 34.41% from an RDT was observed and in Yaoundé with an overall prevalence of 6.5% ,24. A siPlasmodium infections was observed [Plasmodium compared with younger children [Based on the PCR, a high prevalence of 75.0% was observed compared with a similar study carried out in Ghana where a prevalence of 36.8% of asymptomatic observed . A similobserved  with a pchildren . Overallchildren . The trechildren .Plasmodium carriage showed no significant association with gender in the case of Plasmodium malariae (p = 0.382) but a significant association was observed with Plasmodium falciparum (p = 0.038) and P. ovale (p = 0.382). The total number of species identified was 96, which was slightly than the 95 positive sample diagnoses by the RDT. This difference is due to the fact that one individual was found to be harboring more than one species. The average parasitemia was not significantly associated with the prevalence of parasitemia in either gender or the age group. However, other studies [The distribution of the  studies ,35 have  studies . The sen studies ,20, 37. Plasmodium falciparum was the predominant species followed by Plasmodium malariae and then Plasmodium ovale. Thus, understanding the malaria burden in children has a great implication on the interruption of malaria transmission. Ultimately, an accurate, prompt and cheap diagnosis is a key step in managing and treating the disease.Our study identified 85 children positive by a PCR, 95 children positive by an RDT and 71 by LM, revealing a malaria prevalence of 66.9%, 74.8% and 55.9%, respectively.


Background: Normal pressure hydrocephalus (NPH) is characterized by the triad of dementia, gait disturbance and urinary incontinence, all potentially reversible following a ventriculoperitoneal shunt (VPS). This study aims to evaluate the clinical outcomes of shunting in normal pressure hydrocephalus following a new standardized protocol. Methods: This study is designed according to the STROBE guidelines. Demographical, clinical, surgical and radiological data were collected from May 2015 to November 2019. Gait, balance and incontinence data based on the NPH European scale were collected before and after one, six and twelve months of treatment with a VPS. Clinical symptoms and changes of the stoke volume, measured on phase-contrast MRI, were used to evaluate improvement after VPS surgery. Results: One hundred and eighty-one consecutive patients met the inclusion criteria. The mean age was 73.1 years (59–86) and mean follow-up was 38.3 months (13–50). The gait , the balance , continence domain  scores and neuropsychological scales showed a statistically significant improvement over the follow-up. The overall improvement after 12 months was present in 91.2% of patients. An overall complication rate of 8.8% and a reoperation rate of 9.4% were recorded, respectively. Conclusions: Surgical treatment by VPS for NPH improves symptoms in most patients, when accurately selected. A standardized protocol and a multidisciplinary team dedicated to this disorder is needed to achieve an early and correct diagnosis of NPH. Follow-up with stroke volume measurement is a valuable tool for the early diagnosis of shunt malfunction or the need for valve adjustment. Normal pressure hydrocephalus (NPH) is typically characterized by the triad of dementia, gait disturbance and urinary incontinence, all potentially reversible following a ventriculoperitoneal shunt (VPS). Nevertheless, difficulties in differential diagnosis with neurodegenerative diseases result in a delay and/or lack of treatment, thus preventing recovery with dramatic consequences in terms of clinical deterioration and growing socio-economic costs ,2.Pathophysiology of NPH is still debated. Despite the mechanical nature suggested by the name, recent studies focused on periventricular white matter and its vascular supply degeneration as a possible cause ,4. NPH dHowever, the diagnosis of NPH is complicated by the inconsistency that occurs in its clinical presentation and progression. NPH can be similar, or occur in combination with, various diseases that are prevalent in the elderly, such as neurodegenerative disorders , cerebrovascular disease, urological disorders, cervical or lumbar stenosis, and other diseases ,8,9,10.The European Multicenter Study on NPH reported a good correlation between a newly developed assessment scale and the clinical improvement, providing a promising tool for the diagnostic process ,12. SeveThis multicenter prospective observational study aims to evaluate the clinical outcomes of shunting in normal pressure hydrocephalus following a new uniform protocol.This is an observational study conducted at three institutions and approved by the IRB. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement was the checklist for cohort studies that was used to define the study design.Patients with NPH who underwent a ventriculoperitoneal shunt (VPS) between May 2015 and November 2019 at three institutions were prospectively considered for eligibility in the present investigation. A multidisciplinary team, including neurosurgeons, neurologists, neuropsychiatrists and neuroradiologists, validated these inclusion criteria: at least one of the symptoms of NPH triad ; radiological signs on magnetic resonance imaging (MRI) of communicating hydrocephalus ; and the improvement of ≥5 points in the NPH scale, as described by Hallstrom et al. , after aThe exclusion criteria were: other neurodegenerative disorders (excluding mild Alzheimer’s and Parkinson’s disease); secondary hydrocephalus; congenital and/or acquired neurological deficits; obstructive hydrocephalus; Karnofsky performance status (KPS) <60; and less than 12 months of follow-up. A flowchart of the study design is represented in Demographical and clinical data were collected. The total score on the NPH scale, as described by Hallstrom et al., is the summed score of assessments in 4 domains, gait, neuropsychology, balance and continence, divided by 5 (gait has a double weight) or the number of available domain scores. All scores were converted to a min–max range from 0 to 100, where 100 is the performance of an age-matched healthy population. All domains and scores are summarized in The different clinical domains were administered at one time by one neurologist in an outpatient visit, after the 1 day ELD test and at 1, 6 and 12 months after VPS surgery. Preoperative imaging, immediately a postoperative CT scan and MRI scan at 1, 6 and 12 months were also evaluated. 2O.All patients underwent a standard VPS procedure. In addition, before the insertion, multiple slits are created every 4 cm along the catheter in its intraperitoneal segment to create communications between the various compartments of the peritoneum crossed by the catheter. This makes a CSF distribution along the peritoneum compartments possible without accumulation in only one, avoiding pseudocyst formation . The Codt-test. A statistical comparison of the categorical variables was performed by the chi-square statistic using Fisher’s exact test (2-sided). The differences were considered significant at p < 0.05. Statistical analyses were conducted using StatView version 5 software .A statistical comparison of the continuous variables was performed by Student’s A total of 195 patients underwent VPS surgery for NPH at our neurosurgical departments in the investigation period: 14 (7.2%) patients were excluded because they were lost during follow-up for different reasons, and 181 (92.8%) patients matched the inclusion criteria and were finally included in the present investigation. The mean age of the included patients was 73.6 years ± 6.7 (range 59–86), and there were 97 (53.6%) women and 84 (46.4%) men. The mean follow-up was 38.3 months ± 17.3 (range 13–67). The most common symptoms reported were gait disturbance in 173 patients (95.6%), cognitive impairment in 158 (87.3%) and urinary incontinence in 130 (71.8%). The triad of symptoms was present in 128 patients (70.7%). The median preoperative length of symptoms was 21 months (mean 21.3 ± 7.6).p = 0.0003) postoperatively. The demographical and operative data are summarized in The most common co-morbidity was cardiovascular diseases (60.2%), followed by diabetes mellitus (31.5%), obesity (24.9%) and respiratory disorders (21.5%). A total of 56 (30.9%) patients were smokers. A total of 44 patients were in ASA class II (24.3%), 126 (69.6%) in class III and 11 (6.1%) in class IV. The Codman–Hakim valve system was implanted in 49 (27.1%) patients, and the ProGav 2.0 in 132 (72.9%). The mean length of surgery was 44 min ± 11.4 (range 30–90 min), mean blood loss was 60 mL ± 16.5 (range 40–110), mean length of hospitalization was 2 (range 2–5) days, and every patient was mobilized within 24 h after surgery. No intraoperative complications were reported. The mean Evans’ index was 0.39 ± 0.11. The mean stroke volume was significantly reduced from 103.2 µL ± 25.4 preoperatively, to 94 µL ± 22.8 (p < 0.001), the mean balance domain improved from 66.7 (±21.5) preoperatively to 72.4 (±19.2) at last FU (p = 0.001), and the mean continence domain improved from 69.9 (±20.5) preoperatively to 76 (±22) at last FU (p = 0.002). The neuropsychological domain improved from 57 (±12) to 60.2 (±13) (p < 0.011). The mean functional scale score changed from 57.2 (±7.4) to 70.4 (±5.9) at last FU (p < 0.05), and the mean symptomatic scale score improved from 34.5 (±6.8) to 17.7 (±4.5) (p < 0.05). The overall improvement after 12 months was present in 91.2% (165) of patients. Five patients died during follow-up for other reasons not related to NPH or surgery. The clinical outcomes are summarized in The mean gait domain improved from 58.5 (±14.3) to 70.1 (±13.4) at last FU  was decided based on clinical symptoms and changes of the stroke volume on MRI. The new lower pressure was set to enhance the clinical improvement in cases with worsening symptoms and in case of increasing stroke volume measures on follow-up MRIs.Changes in the valve pressure were performed for 169 patients (93.4%) during follow-up. The first opening pressure was 110 mm H2O; 2 patients (1.1%) had a postoperative intraparenchymal hemorrhage that spontaneously recovered within 3 weeks; 2 patients (1.12%) had a superficial wound infection medically treated with oral antibiotic therapy and recovered in 2 weeks; and 4 patients (2.2%) had a postoperative incisional hernia that required a revision surgery.A total of 16 complications were recorded : 8 patients (4.9%) had a subdural hematoma due to over-drainage that spontaneously recovered after setting the opening pressure at 180 mm HA total of 17 patients (9.4%) required surgical revisions for shunt malfunction; 3 patients  presented with proximal catheter obstruction; 7 patients  presented with a non-functioning valve system; and 7 patients  presented with peritoneal catheter extrusion. The complication and reoperation rates are summarized in p = 0.01), stroke volume (p = 0.024), and length of preoperative symptoms before surgery (p < 0.001) independently influenced the clinical outcomes, providing lower chances of responsiveness to surgery. However, a multivariate analysis (p = 0.0001) and duration of the preoperative symptoms (p = 0.037) should be considered as independent factors for non-responsiveness to shunting in this cohort of patients.Subgroup analysis was conducted for investigating how single factors may independently influence clinical outcomes. In the univariate analysis, the Evans’ index  a multidisciplinary team focused on this disease and a uniform protocol is needed to accurately select patients and achieve a correct diagnosis of NPH; (2) surgery with VPS improves gait, balance and continence domain and some of the neuropsychological domain with an impact on quality of life and daily activities of patients; (3) the efficacy of shunting occurred within 3 months after surgery and persisted with a slight reduction at 12 months follow-up in 90% of patients; (4) the one-day ELD showed a high sensibility to select patients with NPH; (5) the use of programmable valves with the anti-siphon system reduce the complication rate related to over drainage; and (6) clinical improvement and the stroke volume measures are necessary to determinate the valve adjustment during follow-up and for the management of NPH in a long-term period.In our study, as previously reported in other studies ,18,19,20After 1 year, 91.2% of patients had an improvement of clinical outcomes after shunting and our results are consistent with the previous studies reporting the clinical benefit of shunt, ranging from 31% to 96% ,19. TomaIn our study, the length of the preoperative symptoms was a significant outcome predictor, as reported in other investigations ,31. A reOf interest, in responders to VPS, we found a postoperative reduction in stroke-volume values on MRI. In our opinion, stroke volume should be considered not only as an outcome predictor, but is also crucial during follow-up for valve adjustments . To the The 91.2% response rate at the last follow-up (mean: 38 months) in the present investigation is attributed to our selection criteria, which include clinical, radiological, and the one-day ELD test. In our opinion, the one-day ELD and the stroke volume measures preoperatively and during follow-up are the key factors that correctly identify the shunt responders’ patients. The one-day ELD test is a reliable tool in NPH diagnosis and predicting patients that will have an improvement in clinical outcomes after shunting surgery, with a sensitivity, specificity and accuracy of 100%, 75.0% and 97.1%, respectively . HoweverThe stroke volume, measured on phase-contrast cine MRI, seems to increase between the onset of the symptoms and the following 18 months, then there is a plateau phase for another 18–20 months, and finally it decreased. However, after its decrease, the patient’s clinical symptoms progressively deteriorate. The progressive reduction in the stroke volume in untreated patients with deteriorating clinical symptoms may be a sign of a progressive cerebral ischemic injury, which renders the NPH symptoms irreversible ,40. The Certainly, the goal of diagnosis is to appropriately refer patients with NPH for shunt surgery and to correctly identify those who do not have NPH or will be non-responders, and suggest against shunting, thus accordingly minimizing the undiagnosed NPH patients or the risks of unnecessary surgery.There are some limitations to be disclosed. Firstly, the patient cohort is small and follow-up is not too long, and certain complications and outcomes could have been consequently unrecognized. Secondly, our results should be validated in other cohorts using similar clinical and radiological measures, and comparative studies of different diagnostic tools are necessary. Finally, this study does not have a control group, which results in lower levels of evidence. Further randomized studies need to be performed with a larger cohort and longer follow-up to validate our findings.Surgical treatment by VPS for NPH improves symptoms in most patients, when accurately selected. Shunting has a low complication rate and guarantees clinical improvements postoperatively for >90% of patients. Appropriate diagnostic procedures and clinical tests that result in accurate patient selection, are the key factors for an acceptable outcome after VPS. At last, careful clinical follow-up and stroke volume measurement on MRI are necessary for valve adjustments to optimize the long-term success of shunt therapy in NPH. Further randomized and comparative clinical trials would better clarify the role of diagnostic procedures on early diagnosis and its theorical impact on long-term clinical outcomes.


There was an error in the original publication . In patiGCDH  , confirming the diagnosis of GA-1.Section 2. Case Reports, 2.2. Case 2, Paragraph 3: At the age of 10 years, genetic testing revealed previously described compound heterozygous variants in GCDH: p.Arg257Gln and p.Met405Val), she was admitted during an infection with poor oral intake, and a clear deterioration of the movement disorder was observed.Section 2. Case Reports, 2.3. Case 3, Paragraph 1: After the diagnosis was made  with 0% residual activity [11] and a milder mutation  with 4–25% residual activity that is more prevalent in African patients [12].The authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected. The original publication has also been updated.


Danio rerio) as a model organism. The zebrafish were exposed to 0.87 mg/L , 1.75 mg/L (5% of 96 h LC50) and 3.51 mg/L (10% of 96 h LC50) of sulfoxaflor for 24 h–48 h and 96 h periods. AChE enzyme activities were analysed by a spectrophotometric method in the brain and muscle tissues. The results of this study showed that in vivo acute sulfoxaflor exposure significantly increased AChE enzyme activity in the brain and muscle tissues of zebrafish. The induction percentages of AChE were between 10 and 83%, and 19 and 79% for brain and muscle tissues, respectively. As a result, it was found that sulfoxaflor had an effect on AChE enzyme activity in the two main tissues containing this enzyme, and it can be considered as a potential neuroactive compound for zebrafish.Sulfoxaflor is the first member of the neonicotinoid-sulfoximine insecticides that acts as an agonist of nicotinic acetylcholine receptors (nAChRs). This study investigated the acute effects of sulfoxaflor on acetylcholinesterase  enzyme activity in the brain and muscle tissues of zebrafish ( Sufoxaflor[methyl(oxo){1-[6-(trifluoromethyl)-3-pyridyl]ethyl}-λ6-sulfanylidene]cyanamide (IUPAC)  is one of the newly developed neonicotinoid-sulfoximine insecticides . SulfoxaThe use of neonicotinoids has increased in the global market during the last two decades ,11,12. SThe neurotoxicity potential of pesticides can be determined by alterations in the cholinesterase (ChE) activities in the different tissues of organisms ,28. ChEsDanio rerio) express AChE in the brain and muscle tissues, and are selected as a model organism in this research, with no detectable butyrylcholinesterase (BChE) activity [Zebrafish  [A commercially available sulfoxaflor {1-[6-(trifluoromethyl)-3-pyridyl]ethyl}-λ6-sulfanylidene]cyanamide) formulation called Transform 500 WG (50% taurine)  was obtaD. rerio) of the wild type  (0.58 ± 0.12 g weight) were commercially supplied. Fish were maintained for 2 weeks in a renewal static system in 100 L glass aquaria with a 14 h/10 h light/dark cycle at 28 °C ± 1 °C during the adaptation periods. Aquaria water was continuously aerated using a static pump system. The physiochemical properties of aquaria water  were recorded regularly. Stock fish were fed with commercial fish pellets twice a day during the adaptation periods. Feeding was stopped 24 h before the toxicity test. Experimental procedures were conducted in accordance with the protocols approved by the Ethics Committee of the Çukurova University Faculty of Medicine Experimental Medicine Research and Application Centre . All toxicity tests were performed with a renewal static system in accordance with the American Public Health Association’s guidelines [Adult mixed sex zebrafish (50) value of sulfoxaflor. The whole experiment was replicated 2 times so that each group was assigned 24 fish. Preliminary tests were conducted to determine the average dose range. One aquarium was set as a control, and 5 different nominal sulfoxaflor concentrations  were applied to the other aquaria. The experiments were conducted for 96 h. Dead fish were recorded per group during the test procedures. At the end of the 96 h exposure period, the LC50 value of sulfoxaflor was determined as 35.13 mg/L , 32.469–38.298) by using probit analysis.Randomly selected fish were divided into 6 groups (1 control group and 5 treatment groups). There were 12 fish in each group. Each group was transferred into 20 L individual glass aquaria in order to determine the 96 h 50% lethal concentration (LCN = 6). Sulfoxaflor concentrations were selected considering toxicity symptoms such as loss of balance, erratic swimming and rapid gill movement based on preliminary tests. In preliminary tests, fish were exposed to 5 different concentrations of sulfoxaflor for 96 h and toxicity symptoms were observed after exposure to >3.51 mg/L (10% of 96 h LC50) sulfoxaflor without mortality. Three sublethal concentrations were chosen, 0.87 mg/L (2.5% of 96 h LC50), 1.75 mg/L (5% of 96 h LC50), 3.51 mg/L (10% of 96 h LC50), for the acute toxicity test. Group I was held in clean water as a control. Groups II, III and IV were exposed to 0.87 mg/L (2.5% of 96 h LC50), 1.75 mg/L (5% of 96 h LC50) and 3.51 mg/L (10% of 96 h LC50) of sulfoxaflor for 24 h–48 h and 96 h. Fish were removed from the aquaria at the end of each test period. Fish were weighed and quickly euthanized by decapitation. The brain and muscle tissues of the fish were carefully dissected out on an ice plate before washing them with saline, weighing and storing them at −80 °C until the analysis was completed.Acute toxicity tests were carried out in 4 separate 60 L glass aquaria. The selected fish were randomly divided into 4 different experimental groups . Each experimental group comprised 108 individuals: 36 fish for the 24 h exposure period, 36 fish for the 48 h exposure period and 36 fish for the 96 h exposure period. Furthermore, each experimental unit consisted of a pool of 6 individuals, from which the required amount of tissue samples was collected and pooled for each individual measurement. Thus, the measurements were repeated in 6 technical replicates . The homogenates were centrifuged  at 13,000× AChE activities in brain and muscle tissues were detected by using the spectrophotometric method  according to the modified method  developeThe protein levels of homogenates obtained from brain and muscle tissues were determined by utilizing the method developed by Bradford . For thi50 value was determined by probit analysis using the SPSS 22.0 package program. All data are denoted as means ± standard error. Analysis of variance (one-way ANOVA) and Duncan’s multiple comparison tests were used to determine statistical differences between the control and test groups using the SPSS 22.0 package program.The LC50 of sulfoxaflor was calculated as 35.13 mg/L for zebrafish following probit analysis , 32.469–38.298, p < 0.05)  .p < 0.05). The results indicated that the 96 h exposure period had a greater effect on AChE enzyme activity compared with other exposure periods in the brain. AChE enzyme activities rose significantly with higher concentrations of sulfoxaflor in the brain (p < 0.05). The activity of AChE enzymes decreased by 17% with 0.87 mg/L sulfoxaflor exposure and increased by 10% and 25% with 1.75 mg/L and 3.51 mg/L sulfoxaflor exposure at 24 h, respectively (p < 0.05). In addition, inductions in AChE activity were determined at 96 h for all tested sulfoxaflor concentrations. AChE enzyme activity was induced by 27%, 29% and 83% with 0.87 mg/L, 1.75 mg/L and 3.51 mg/L, sulfoxaflor exposure at 96 h, respectively (p < 0.05).Sulfoxaflor significantly increased the activity of AChE enzymes in the brain of zebrafish . The results indicated that AChE enzyme activity was significantly increased by high concentrations of sulfoxaflor during all of the exposure periods in the muscle (p < 0.05). Sulfoxaflor exposure significantly increased AChE enzyme activity by 19% for 3.51 mg/L concentration at 24 h (p < 0.05). Similarly, AChE enzyme activity was induced by 21% and 24% with 1.75 mg/L and 3.51 mg/L sulfoxaflor exposure at 48 h, respectively (p < 0.05). The activity of AChE enzymes was increased by 53%, 34%, and 79% by 0.87 mg/L, 1.75 mg/L and 3.51 mg/L sulfoxaflor exposure at 96 h, respectively (p < 0.05). The results demonstrated that the elevations in AChE enzyme activity were not dependent on the sulfoxaflor concentrations and exposure periods in the muscle  sulfoxaflor without mortality. It can be concluded that sulfoxaflor may cause similar results on animal behaviour, considering previous studies that indicated a relationship between the changes in AChE enzyme activity and the behavioural alterations caused by neonicotinoids. Thus, it is obvious that evaluation of the neurotoxic potential of sulfoxaflor with its effect on behavioural changes in fish and nontarget animal species is important for risk assessment and environmental health.The tests related to the effects of pesticides on behavioural alterations in animals are necessary for evaluating their neurotoxic effects and their effects on the endocrine system . These bNeonicotinoids have highly variable environmental half-lives ranging from minutes to several weeks in water . TherefoThe possible risk of environmental contamination due to the increasing use of neonicotinoid insecticides can create problems for human, animal and environmental health. Although there is clear evidence of the effects of other neonicotinoids on nontarget animal species in the literature, a limited number of studies are available related to the in vivo effects of sulfoximine containing neonicotinoids on other nontarget species. The present study demonstrated that sulfoxaflor acutely caused inductions of AChE enzyme activity in the brain and muscle tissues of zebrafish, in contrast to most of the other neonicotinoids. Thus, the inductions in AChE caused by sulfoxaflor might indicate that sulfoxaflor can be considered as a neuroactive compound for zebrafish. In addition, the tissue-specific response was not determined due to similar induction rates of AChE in both tissues. AChE induction might not be suggested as a biomarker due to the lack of a dose–response relationship for sulfoxaflor exposure. Many of the neonicotinoids cause behavioural changes and produce a neurotoxic response in fish and mammals through AChE inhibition. These findings highlighted the possible effects of sulfoxaflor on AChE enzymes in fish. Further studies are needed to clarify the actual toxic effects of sulfoxaflor on the nervous system by investigating main neurotoxicity parameters and behavioural alterations in nontarget animal species.


To investigate the clinical features and prognosis in patients of hyperlipidemic acute pancreatitis with or without diabetes.  157 patients with hypertriglyceridemic pancreatitis (HTGP) were included in this study. Patients with a previous history of diabetes were identified in the group of HTGP with diabetes (HTGPD), while patients without a history of diabetes were identified in the group of HTGP. The clinical characteristics and prognosis data of these patients in the two groups were analyzed. p < 0.001). Compared to patients of HTGP, those of HTGPD had older age of onset, higher blood glucose levels, and higher GSP levels on admission. Electrocardiograms showed that patients of HTGPD had a significantly higher risk of heart ischemia than those of HTGP (p < 0.05). Patients of HTGPD had higher APACHE II scores than those of HTGP (p < 0.001). Single-factor analysis showed that higher triglyceride levels, GSP, LDL, and previous history of diabetes were associated with HTGP recurrence.  Multivariate Cox regression analysis showed that age, body mass index, glycated serum protein (GSP), and Acute Physiology and Chronic Health Evaluation (APACHE) II score were significantly associated with mortality in patients with HTGP. The mortality was significantly higher in the HTGPD group than in the HTGP group ( Clinicians should be alert to patients of HTGP with diabetes. Diabetes is an important risk factor for HTGP and hyperglycemia may affect the development and prognosis of HTGP. Diabetes has become endemic in China. Based on the most recent data, 12.8% of the population in China has diabetes, while as much as 35.2% of the population has prediabetes . In otheThis was a retrospective study over 3 years from January 2010 to December 2012. 157 patients with HTGP were diagnosed and treated at The First Affiliated Hospital of Nanchang University. The diagnosis of HTGP required (1) diagnosis according to American guidelines of the diagnosis and treatment of AP . Patientt-test, the Mann-Whitney U-test, and the chi-square test were used to describe patients' baseline characteristics. Survival curves were plotted according to the Kaplan-Meier method, and their statistical differences were analyzed by the log-rank test. Cox proportional hazard regression was used to test the association between clinical prognostic factors and mortality in patients with HTGP. Variables that showed an indication of prognostic influence in univariate Cox regression analysis were included in multivariate Cox regression analysis using the backward elimination method. All the differences in the clinical features were analyzed between HTGP and HTGPD. Logistic regression was used to analyze the risk factors of HTGP recurrence. All statistical analyses were performed using the SPSS Statistical Package (version 20.0). Values of p < 0.05 were considered statistically significant.Normally distributed data were expressed as means ± SD, whereas variables with a skewed distribution were reported as median (interquartile range) and log-transformed to approximate normality before the analysis. Enumeration data were expressed as a positive rate or percentage. Student's p < 0.001). Disease incubation period occurred slightly later in the HTGPD group than in the HTGPA group. The patients in the HTGPA group had a shorter time from first symptoms to admission, and most patients in this group went to the hospital for abdominal pain. Almost all patients in the HTGP group had obvious abdominal pain, while only 81.0% of the subjects in the HTGPD group had abdominal pain. All patients had a higher than normal BMI  and 53 patients with HTGPD. The mean age of the total 157 patients in this study was 47.9 ± 12.7  years; there were more males than females . Among them, 104 had HTGPA, and 53 were HTGPD. All 53 diabetic patients were of type 2. The average diabetic course was 4.0 ± 1.7 years. All of 157 patients had characteristic CT imaging changes of AP. Among the 53 patients with HTGPD, 48 took oral antidiabetic drugs, two underwent diet and exercise intervention, and three were under no treatment. In the HTGPA group, 21 (20.2%) subjects had ischemic heart disease, eight (7.7%) had chronic obstructive pulmonary disease (COPD), and 30 (28.8%) had hypertension. In the HTGPD group, 12 (22.6%) subjects had ischemic heart disease, four (7.5%) had the COPD, and 18 (33.9%) subjects had hypertension. The mean APACHE II score of the HTGPD group (7.1 ± 3.5) was significantly higher than that of the HTGPA group (5.1 ± 3.2) . The mean GSP of the HTGPD group (2.8 ± 0.6 mmol/L) was higher than that in the HTGPA group (1.9 ± 0.4 mmol/L). The mean low-density lipoprotein cholesterol (LDL-C) level of patients in the HTGPD group (2.0 ± 1.2 mmol/L) was higher than that of patients in the HTGPA group (1.1 ± 0.8 mmol/L). The mean CRP level of the HTGPD group (229.2 ± 108.7 mg/L) was higher than that of the HTGPA group (162.7 ± 109.7 mg/L). Electrocardiogram findings revealed that patients with HTGPD had a significantly higher risk of heart ischemia than patients with HTGPA  than those in the HTGPA group (8.6 ± 3.7 mmol/L), (p < 0.001). Univariate analysis showed that triglyceride levels and previous history of diabetes were associated with HTGP recurrence. On stepwise logistic regression analysis, HTG and previous history of diabetes were associated with HTGP recurrence . Survival curves showed a significant survival difference between the HTGPD group and HTGP group via the log-rank test , 18 had HTGPD, meaning that the recurrence rate of patients with HTGPD  was higher than that of patients with HTGPA (10/104; 9.6%; ank test , while mank test .The prevalence of diseases associated with obesity and diabetes has increased in recent years and contributes substantially to healthcare costs and mortality rates . In ChinMost of the patients with HTGP were accompanied by hyperglycemia . AnalysiIn the current study, LDL-C levels of patients with HTGPD were higher than those of patients with HTGPA. A high LDL-C contributes to an elevated cardiovascular risk, thus increasing mortality rates of patients with HTGPD. Controlled LDL-C levels facilitate a substantial reduction in cardiovascular risk . As is sIn conclusion, physicians should be alert to HTGPD. Diabetes is an important risk factor for HTGP, and hyperglycemia may affect the development and prognosis of HTGP.


GPX1 (rs1050450), DIO2 (rs225014) and SEPP1 (rs7579) gene polymorphisms and the development of chronic or neoplastic diseases. However, there are no reports on the influence of these polymorphisms on the development of endometrial cancer. Methods: 543 women participated in the study. The study group consisted of 269 patients with diagnosed endometrial cancer. The control group consisted of 274 healthy women. Blood samples were drawn from all the participants. The PCR-RFLP method was used to determine polymorphisms in the DIO2 (rs225014) and GPX1 (rs1050450) genes. The analysis of polymorphisms in the SEPP1 (rs7579) gene was performed by means of TaqMan probes. Results: There was a 1.99-fold higher risk of developing endometrial cancer in CC homozygotes, DIO2 (rs225014) polymorphism , compared to TT homozygotes. There was no correlation between the occurrence of GPX1 (rs1050450) and SEPP1 (rs7579) polymorphisms and endometrial cancer. Conclusion: Carriers of the DIO2 (rs225014) polymorphism may be predisposed to the development of endometrial cancer. Further research confirming this relationship is recommended.Background: Numerous studies indicate a relationship between the presence of  Endometrial cancer is the most common cancer of the female reproductive system. In 2020, there were over 400,000 new cases of endometrial cancer worldwide, and an increase in the incidence compared to previous years was observed . There aSelenium is a trace element that is part of the selenoproteins. There are 25 selenoproteins in the human body. All of them contain selenium in their active center, which forms the amino acid selenocysteine (Sec) . ScientiGPX1 gene is located on the short arm of chromosome 3; it encodes the glutathione peroxidase 1 enzyme. This enzyme is mainly expressed in red blood cells, liver, lung and kidney tissues. It has a protective function against oxidative stress [GPX1 rs1050450 (Pro198Leu). As a result of this polymorphism, there is a change of cytosine (C) to thymine (T) in codon 198. This leads to the change of amino acids from proline (CCC) to leucine (CTC) [The e stress . The mosne (CTC) .DIO2 gene is located on the long arm of chromosome 14; it encodes the iodothyronine 2 deiodinase enzyme. This enzyme is found mainly in the thyroid gland, brain, pituitary gland, skeletal muscle, heart and skin [The and skin . The maiand skin . The mosand skin .SEPP1 gene is located on chromosome 5 and encodes Selenoprotein P1 [SEPP1 polymorphisms is rs7579, which changes guanine (G) to adenine (A) in the 3′ untranslated region (3′ UTR) of the mRNA [The otein P1 . This protein P1 . It occuthe mRNA .GPX1 gene (glutathione peroxidase 1) and the following neoplasms: breast, prostate, bladder and lungs [SEPP1 gene (selenoprotein P1) with colorectal and prostate cancer [DIO2 gene (iodothyronine 2 deiodinase) was associated with the occurrence of diseases such as: type 2 diabetes, osteoporosis, Hashimoto’s disease, Graves’ disease, Alzheimer’s disease and autism [It has recently been observed that some forms of selenoprotein genes are associated with the risk of several cancers. Such a relationship was observed between the rs1050450 polymorphism of the nd lungs ,15,16,17e cancer ,19. The d autism ,21,22,23The study involved 269 patients diagnosed with endometrial cancer based on a previously performed histopathological examination. These patients reported to the Department of Gynecology and Clinical Obstetrics, Provincial Hospital No. 1, in Rzeszow in 2016–2020 in order to start oncological treatment. Before the planned surgery, blood samples were drawn from each patient. The blood was also collected from 274 healthy women who were included in the control group. All study participants agreed to have blood samples drawn for clinical trials. The research was approved by the Bioethics Committee . All samples were stored at −80 °C until DNA isolation.DNA from test and control groups was isolated from peripheral blood lymphocytes using the Quick Blood DNA Purification Kit . The protocol attached by the manufacturer was followed. The quality and quantity of the isolated genomic DNA was determined using Nanodrop . All samples were standardized by dilution to a final concentration of 25 ng/μL. The samples were stored at −20 °C until further analysis.DIO2 (rs225014) and GPX1 (rs1050450) genes. The DIO2 and GPX1 polymorphisms were amplified using primers published by Xiong et al. The amplification was performed in the final concentration reaction mixture containing 1 × PCR buffer, 0.25 mM dNTP mixture, 1.5 mM MgCl2, Pc µM, each of the forward and reverse primers, 0.5 U Taq DNA polymerase  and 50 ng of purified DNA. The final reaction volume was 20 µL. The PCR reaction conditions were as follows: initial denaturation at 95 °C for 10 min, extension of 35 cycles of denaturation at 95 °C for 30 s, hybridization at 58 °C for 45 s, extension at 72 °C for 45 s and final extension at 72 °C for 7 min. The reactions were performed in a T100™ thermocycler . The PCR products were digested with the appropriate restriction enzymes and run on a 2% Midori Green Stain-stained agarose gel .The PCR-RFLP method was used to determine polymorphisms in the SEPP1 (rs7579) gene was performed by an allele discrimination test using the TaqMan genotyping test . The real-time PCR reaction conditions were as follows: 95 °C for 10 min and 40 cycles, 95 °C for 15 sec and 60 °C for 1 min. The reaction was performed on a Roche Molecular Diagnostics Cobas z480 Analyzer in 96-well plates. The Endpoint Genotyping module  was used for data analysis.The analysis of polymorphism in the In order to estimate the odds ratio (OR) of endometrial cancer depending on the genotype of the analyzed genes, one-way logistic regression models were used.GPX1 gene were p(C) = 0.72 and q(T) = 0.28 in the study group and p(C) = 0.71 and q(T) = 0.29 in the control group, respectively. The allele frequency for the DIO2 gene in the test group was p(T) = 0.61 and q(C) = 0.39, and in the control group, p(T) = 0.69 and q(C) = 0.31, whereas for the SEPP1 gene in the test group, p(G) = 0.71 and q(A) = 0.29 and in the control group, p(G) = 0.72 and q(A) = 0.28; for all genes, p > 0.05.The genotype distribution for all genes was at Hardy–Weinberg equilibrium. The allele frequencies for the GPX1 (rs1050450), DIO2 (rs225014) and SEPP1 (rs7579) in the control and study groups.GPX1 (rs1050450) polymorphism, the CC genotype was found in 139 patients from the control group and in 129 women from the study group. CT heterozygous was found in 108 women from the control group and 124 from the study group. A TT genotype was observed in 27 patients from the control group and 16 from the study group. The OR for the CT heterozygote was 1.24 (95% Cl 0.87–1.76), and for the TT homozygote was 0.64 (95% Cl 0.32–1.23). However, the differences observed were not statistically significant.In the case of the DIO2 (rs225014) polymorphism, 128 patients from the control group and 103 from the study group were TT homozygotes. TC heterozygotes were represented by 121 patients from the control group and 126 from the study group. CC homozygous was found in 25 patients from the control group and in 40 patients from the study group. A statistically significant, more frequent occurrence of the rs225014 polymorphism was found in patients with endometrial cancer compared to patients in the control group . The OR for the CC homozygote was 1.99 (95% Cl 1.14–3.53). The OR for the TC heterozygote was 1.29 (95% Cl 0.90–1.86), without, however, reaching statistical significance .In the case of the SEPP1 (rs7579) polymorphism, the distribution was as follows: 138 women from the study group and 145 from the control group had the GG genotype. A total of 107 women from the control group and 109 from the study group were heterozygous GA. An AA genotype was obtained in 22 patients from the test group and 22 from the control group. The odds ratio for the AA homozygote was 1.05 (95% Cl 0.55–1.99), and for the GA heterozygote was 1.07 (95% Cl 0.75–1.53). However, the results obtained are not statistically significant.In the case of the GPX1 (rs1050450), DIO2 (rs225014) and SEPP1 (rs7579), are associated with the risk of developing endometrial cancer. Significantly, one of the polymorphisms analyzed in the DIO2 (rs225014) gene was more common in patients with endometrial cancer . The results of our study showed that carrying the CC genotype resulted in almost twice the risk of endometrial cancer . In the review of the literature, we did not find any papers with a similar research problem. The presence of TC polymorphism also indicates a trend that may be associated with an increased risk of endometrial cancer (OR 1.29), but these differences did not reach statistical significance   . Our obsDIO2 gene has been observed in follicular, anaplastic and medullary thyroid cancer [DIO2 gene was demonstrated in pituitary tumors and in brain gliomas [DIO2 gene was also noted in skin cancer [DIO2 (rs225014) polymorphism and neoplasms; however, numerous studies indicate a correlation between the DIO2 (rs225014) polymorphism and chronic diseases. In 2010, it was shown that CC homozygote carriers had a higher risk of developing type II diabetes . Meta-analyzes confirmed this relationship—the total effect from four studies indicated an OR of 1.18  [DIO2 (rs225014) polymorphism on the risk of developing type 2 diabetes, as well as on insulin resistance and obesity [p-value 0.003) compared to other genotypes [DIO2 (rs225014) polymorphism on bone mineral density was also investigated. Research shows that people with Thr92Ala polymorphism have a higher risk of developing osteoporosis compared to people with the wild-type genotype [DIO2 (rs225014) polymorphism on the occurrence of autoimmune thyroid diseases (Hashimoto’s disease and Graves’ disease). It was observed that patients with the TT and TC genotypes had an increased risk of developing autoimmune diseases of the thyroid gland, especially Hashimoto’s disease [DIO2 (rs225014) polymorphism increased the risk of developing Alzheimer’s disease (AD)/dementia among African Americans  by 1.3 times. In the population of Americans of European descent, no influence of the DIO2 gene polymorphism on the development of AD or dementia was reported [DIO2 polymorphism (rs225014) and the incidence of endometrial cancer is the first consideration of this type.So far, increased activity of the d cancer ,25. Simi gliomas ,27. Alten cancer . No corrue 0.02) . In 2007 obesity . T. Ota enotypes . The infgenotype ,33. In 2 disease . A. Guer disease . In 2018 [DIO2 rs5014 polyreported . The obsGPX1 (rs1050450) polymorphism has been studied for the following cancers so far: breast, prostate, bladder, lung, head, neck and brain. In 2009, N.A. Ermolenkoa et al. observed that the presence of the T allele (rs1050450) in the GPX1 gene reduces the risk of breast cancer  [p-value 0.035) [GPX1 gene had an increased risk of developing breast cancer [GPX1 (rs1050450) gene had a 1.43 times higher risk of breast cancer compared to patients with the CC genotype [GPX1 (rs1050450) polymorphism had no effect on the development of breast cancer [The e 0.012) . In 2015e 0.035) . In 2010t cancer . G. Ravngenotype . In 2004t cancer .GPX1 (rs1050450) polymorphisms and the risk of developing prostate cancer [In 2011, O. Erdem et al. reported no relationship between e cancer . C. Kucue cancer . In 2013e cancer .GPX1 (rs1050450) polymorphism and the incidence of bladder cancer [p-value 0.019) [GPX1 (rs1050450) polymorphism was associated with the occurrence of bladder neoplasms  [In 2016, K. Hadami et al. showed no correlation between the r cancer . Ch. Wan <0.001) .p-value 0.02 and OR 1.45, 95% Cl 1.17–1.81, p-value 0.02) compared to people without these addictions [The impact of the Pro198Leu polymorphism was also studied to assess the risk of developing colorectal cancer. No such dependency was found. The only thing that was observed was that people with the TT genotype (rs1050450) who smoked cigarettes or drank alcohol had a higher risk of developing colorectal cancer  had a reduced risk of developing lung cancer  [In 2008, A. Rosenberger et al. conducted an analysis that showed that the carriers of the Leu smokers) . Studiessmokers) . The oppsmokers) .A meta-analysis from 2017 showed a connection between the rs1050450 C > T polymorphism and the development of head, neck and brain cancer .SEPP1 (rs7579) polymorphism was analyzed for the colon and prostate. In 2010, C. Méplan et al. reported that people with the nonGG genotype of the SEPP1 (rs7579) polymorphism had a 1.43 times higher risk of developing colorectal cancer compared to patients with the GG genotype . The OR for AA homozygotes was 2.47  [SEPP1 (rs7579) polymorphism had a 1.67 times higher risk of developing colorectal cancer [p-value 0.05) for the presence of the A allele in the rs7579 SEPP1 polymorphism [SEPP1 (rs7579) polymorphism and the development of colorectal cancer [The e 0.019) . In 2013SEPP1 (rs7579) polymorphism, but without statistical significance  [An increased risk of prostate cancer development was observed in patients with the AA genotype  . In 2010ue 0.22) .GPX1 (rs1050450) and SEPP1 (rs7579) and the occurrence of endometrial cancer.However, our analysis did not show a relationship between In addition, in our research, apart from selected polymorphisms, we analyzed the most common risk factors for endometrial cancer: obesity, diabetes and arterial hypertension. We subjected the results to univariate and multivariate analysis.p-value < 0.001) [In 2017, D. Aune et al. conducted a literature review and meta-analysis of the research. From 19 case-control studies and 6 cohort studies, the total relative risk (RR) of developing endometrial cancer in patients with arterial hypertension was 1.61 (95% CI 1.41–1.85) . O. Ragl< 0.001) .p-value < 0.001). Diabetes mellitus in univariate analyses significantly influences the development of endometrial cancer (OR 2.86). However, there is no statistical relevance in multivariate analyses.Hypertension and obesity in both univariate and multivariate analyses increase the risk of endometrial cancer (DIO2 polymorphism (rs225014) for the CC genotype, we obtained an OR of 1.99 in univariate analyses . In multivariate analyses for the same genotype, the OR was 2.06 . For the remaining TT and TC genotypes, statistical significance was not achieved in either univariate or multivariate analyses.In the case of the GPX1 (rs1050450) and SEPP1 (rs7579), statistical significance was not achieved in either univariate or multivariate analyses , DIO2 (rs225014) and SEPP1 (rs7579) and the possibility of developing endometrial cancer. Our analyses show that the DIO2 (rs225014) polymorphism may predispose to the development of endometrial cancer. Due to the fact that no similar studies were found during the literature review, and our studies were conducted on a limited number of patients, further analyses are recommended to confirm the relationship between the DIO2 (rs225014) polymorphism and the development of endometrial cancer.In the literature review, no analyses were found regarding the relationship between the polymorphisms