Objective:The degree of involvement of extraocular muscles varies across different regions of retrobulbar tissue in patients with thyroid eye disease, but the mechanism is unclear. This study aims to explore the relationship between differential expression of thyroid-stimulating hormone receptor(TSHR) in different parts of the extraocular muscles and the varying degrees of muscle involvement.Methods:The medial, lateral, superior, and inferior rectus muscle were separated from the retrobulbar tissue of rats, and the expression level of TSHR in four extraocular muscles was detected by immunofluorescence and qPCR. Extraocular muscle tissue of patients with strabismus was collected to detect the expression of TSHR and the cell types expressed by fluorescence.Results:The results of qPCR showed that the expression of TSHR in the medial rectus muscle was significantly higher than that in the lateral, superior, and inferior rectus muscle(medial rectus vs lateral rectus, P=0.012; medial rectus vs superior rectus, P=0.015; medial rectus vs inferior rectus, P=0.013), but there was no difference in insulin-like growth factor 1(IGF-1R) expression. Immunofluorescence showed that TSHR was co-expressed with PAX7, a molecular marker of muscle satellite cells, and the expression level in the medial rectus muscle of rats and humans was significantly higher than those in the other three extraocular muscles. Conclusion:The high specific expression of TSHR in the satellite cells of the medial rectus muscle may be the reason why the medial rectus muscle is most susceptible to involvement in thyroid eye disease.

Objective:To explore clinical value of nucleic acid detection for hepatitis B virus (HBV) screening in hospitalized patients.Methods:This cross-sectional study collected and analyzed plasma samples from patients admitted to 10 domestic medical institutions from July 2021 to December 2021. Serological immunoassay and nucleic acid screening were used to simultaneously detect hepatitis B markers such as hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B e Antigen (HBeAg), hepatitis B e antibody (HBeAb), hepatitis B core antibody (HBcAb),and HBV DNA. Statistical analysis was performed on the serology, nucleic acid test results and clinical information of the patients.Results:Of the 8 655 collected samples, HBsAg was positive in 216 (2.50%) samples,HBV DNA was positive in 238 (2.75%) samples ( P>0.05); 210 (2.43%) samples were positive for both HBsAg and HBV DNA, 28 (0.32%) were HBsAg negative and HBV DNA positive, 6 cases (0.07%) were HBsAg positive and HBV DNA negative. Conclusion:These results indicate that the HBV DNA testing is equally effective as hepatitis B virus serological detection for hepatitis B virus screening in hospitalized patients.

Objective:This multi-centre study was conducted to assess the efficacy of various preoperative/pre-transfusion screening methods for blood transmitted disease.Methods:From July 2021 to December 2021, plasma samples of patients admitted to 10 hospitals were collected for screening preoperative/pre-transfusion blood transmitted disease. Nucleic acid detection technology was used to detect hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA and human immunodeficiency virus (HIV)(1+2) RNA, and the results were compared with the immuno-serological methods. χ 2 test and Kappa test were used to analyze the efficacy of these two methods. Results:A total of 8 655 valid specimens were collected from 10 hospitals. There was a statistically significant difference in the positive detection rate of HCV between the two methods ( P<0.001). There was no significant difference in the positive detection rate of HBV and HIV assessed by the two methods ( P>0.05), but the number of positive cases detected by HBV DNA and HIV RNA (218 and 4 cases) was significantly higher than the corresponding serological results (216 and 2 cases). At the same time, there were HBV, HCV and HIV immuno-serological omissions by the immuno-serological methods, among which 28 cases were HBsAg negative and HBV DNA positive, 2 cases were HCV antibody negative and HCV RNA positive, and 2 cases were HIV antigen/antibody negative and HIV RNA positive. In addition, in the 66 samples with inconsistent results from the two detection methods, 83.3% (55/66), 68.2% (45/66), 63.6% (42/66) and 62.1% (41/66) of patients aged was>45 years, tumor, surgery and male, respectively. Conclusions:Compared with immuno-serological tests, nucleic acid tests have the advantage in terms of sensitivity on detecting HBV, HCV and HIV infection and could reduce missed detection. The risk of transmission can be reduced by adding HBV, HCV, and HIV nucleic acid tests to preoperative/pre-transfusion immuno-serological tests screening for patients over 45 years of age and tumor patients.

Objective:To compare the cost-effectiveness of hospitalized Chinese patients undergoing nucleic acid screening strategies for hepatitis B and hepatitis C, immunological screening strategy, and no screening strategy under different willingness to pay (WTP). The results might aid to decision-making for the optimal strategy.Methods:In this study, nucleic acid screening, immunological screening and no screening were used as screening strategies, and China′s GDP in 2021 (80 976 yuan) was used as the threshold of WTP to construct a Markov model. After introducing parameters related to the diagnosis and treatment of hepatitis B and C in inpatients, a cohort population of 100 000 inpatients was simulated by TreeAge Pro 2021 software, the total cost, total health effects, incremental cost-effectiveness ratio and average cost-effectiveness ratio of different screening strategies were calculated, and cost-effectiveness analysis was conducted. Univariate and probabilistic sensitivity analysis were used to assess the impact of parameter uncertainty on the final results.Results:Compared with the non-screening strategy, the incremental total cost of the hepatitis B immunological screening strategy for cohort patients was 11 049 536 yuan, and the incremental cost-effectiveness ratio was 24 762 yuan/quality-adjusted life years (QALY), while the total incremental cost of nucleic acid screening was 19 208 059 yuan, and the incremental cost-effectiveness ratio was 29 873 yuan/QALY; the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 834 yuan/QALY. Compared with the non-screening strategy, the incremental cost-effectiveness ratio of hepatitis C immunological screening strategy was 5 731 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening strategy was 8 722 yuan/QALY, the incremental cost-effectiveness ratio of nucleic acid screening and immunological screening was 45 591 yuan/QALY. The results of probabilistic sensitivity analysis showed that when the cost of nucleic acid testing exceeded 214.53 yuan, it was not cost-effective to perform hepatitis B nucleic acid screening under the WTP as 1 fold GDP. When the cost of nucleic acid testing exceeded 132.18 yuan, it was not cost-effective to conduct hepatitis C screening under the WTP as 1 fold GDP.Conclusions:Nucleic acid screening strategy can achieve more cost-effectiveness and is worthy of vigorous promotion. Compared with no screening, both the nucleic acid and immunological screening strategies are cost-effective, and hepatitis nucleic acid screening is the optimal strategy for hospitalized patients.

Objective To investigate the mutation characteristics and influencing factors of ethambutol(EMB)resistance gene of Mycobacterium tuberculosis in Guangxi Zhuang Autonomous region,and to provide evidence for molecular diagnosis and clinical treatment of tuberculosis.Methods A total of 655 strains of Myco-bacterium tuberculosis(52 ethambutol resistant strains and 603 ethambutol sensitive strains)were collected continuously from 30 TB drug resistance monitoring sites in Guangxi in 2018-2019,and the mutation characteristics and influencing factors of ethambutol resistant genes were analyzed by whole genome sequencing.Results Among 655 strains of Mycobacterium tuberculosis,54 strains had ethambutol drug resistance gene mutation,the mutation rate was 8.24%(54/655).Among 52 EMB-resistant strains detected by proportional method,21 had gene mutation,the mutation rate was 40.38%(21/52),and 33 of 603 EMB-sensitive strains had gene mutation,the mutation rate was 5.47%(33/603).The gene mutation rate in drug-resistant strains was higher than that in sensitive strains(χ2 = 77.133,P = 0.000).The coincidence rate of EMB drug resistance phenotype and gene mutation was 40.38%(21/52),and the results of the two tests were not highly consistent(Kappa = 0.343,P<0.001).The mutant genes of 54 strains were embA,embB and embC,and there were 20 mutant forms,among which 29 were mutated at unit point,accounting for 53.70%(29/54),and 25 were mutated at joint site,accounting for 46.30%(25/54).Among the unit point mutations,embB306(35.19%)had the highest mutation proportion,followed by embB497(5.56%)and embB406(3.70%).Among the joint site mutations,embC270+embB378 had the highest mutation proportion(22.22%),The second was embB306+embA-12(3.70%).Gender,anti-tuberculosis treatment history,genotype and MDR might be related to EMB gene mutation(χ2 = 9.388,P = 0.004;χ2 = 27.084,P = 0.000;χ2 = 6.671,P = 0.010;χ2 = 68.826,P = 0.000).Multivariate logistic regression analysis showed that male(OR = 6.150),retreatment(OR = 2.636)and multidrug resistance(OR = 7.333)may be risk factors for EMB resistance gene mutation,and Beijing genotype may be a protective factor for EMB resistance gene mutation(OR = 0.511).Conclusion EMB resistance of Mycobacterium tuberculosis is related to embA,embB and embC gene mutations,and the incidence of EMB resistance phenotype is not high.For male,retreatable,MDR-resistant,and non-Beijing genotype TB patients,attention should be paid to the mutation of the EMB resistance gene.

Objective:To investigate the expressions of miRNA-4686 (miR-4686) and protein disulfide isomerase A4 (PDIA4) in esophageal cancer tissues, and the effect of miR-4686 on the proliferation and migration of esophageal cancer cells in vitro and possible mechanisms.Methods:The complementary integration of miR-4686 and PDIA4 mRNA were predicted using the miRWalk version 3 online website. The expression profiles of miR-4686 and PDIA4 mRNA were obtained from the University of California, Santa Cruz Genome (UCSC) database, and the relative expression of miR-4686 and PDIA4 mRNA in esophageal cancer tissues and paracancerous tissues were analyzed. The UCSC database was used to obtain the localization of miR-4686 in human cells. The relative expressions of miR-4686 and PDIA4 mRNA in esophageal cancer Eca109, TE-13, EC9706 and KYSE-510 cells and normal esophageal epithelial HET-1A cells were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Eca109 cells with the lowest relative expression of miR-4686 were selected for subsequent research, and the Eca109 cells were divided into miR-4686 group (transfected with miR-4686 mimic) and miR-NC group (transfected with miR-4686 negative control sequence mimic). The proliferation ability of Eca109 cells in the two groups was detected by colony formation assay, the migration ability of Eca109 cells was detected by scratch healing assay, and the targeting relationship between miR-4686 and PDIA4 mRNA was verified by dual luciferase reporter gene assay; the expressions of PDIA4 protein and PI3K-AKT-mTOR signaling pathway-related proteins were detected by Western blotting.Results:Compared with the paracancerous tissues, miR-4686 expression was low and PDIA4 mRNA expression was high in esophageal cancer tissues (both P < 0.01). Compared with normal esophageal epithelial HET-1A cells (0.98±0.15), the relative expression of miR-4686 in esophageal cancer Eca109 (0.11±0.04), TE-13 (0.58±0.10), EC9706 (0.34±0.05) and KYSE-510 cells (0.69±0.06) were all decreased (all P < 0.05). Compared with the miR-NC group, the relative expression of miR-4686 in Eca109 cells of the miR-4686 group increased (9.4±2.1 vs. 1.0±0.4, t = 3.88, P = 0.008), the number of colony formation decreased (38±9 vs. 114±18, t = 3.78, P = 0.009), the scratch healing rate decreased [(27.13±0.91)% vs. (45.05±3.89)%, t = 4.48, P = 0.004], and the relative expression of PDIA4 mRNA decreased [1.0±0.5 vs. 6.3±0.9, t = 5.04, P = 0.002]. Compared with wild type (WT)-PDIA4+miR-NC group, the relative luciferase activity of Eca109 cells in WT-PDIA4+ miR-4686 group decreased (0.31±0.08 vs. 0.99±0.08, t = 5.96, P < 0.001). Compared with miR-NC group, the expressions of PDIA4, p-PI3K, p-AKT and p-m-TOR proteins in Eca109 cells of the miR-4686 group were all decreased. Conclusions:In esophageal cancer tissues, miR-4686 is lowly expressed and PDIA4 mRNA is highly expressed. miR-4686 may inhibit the proliferation and migration of esophageal cancer Eca109 cells by targeting regulation of PDIA4 expression.

Background::Essential tremor (ET) and Parkinson’s disease (PD) are common movement disorders. ET-PD syndrome is characterized by the occurrence of PD in patients with a previous history of ET, which may be an independent phenotype distinct from PD. This study aims to identify clinical characteristics and subtypes in ET-PD.Methods::A total of 93 newly diagnosed ET-PD patients and 93 newly diagnosed PD patients matched for age, sex, education, and disease duration of PD were selected using propensity score matching analysis. The K-means cluster analysis was performed for 11 variables derived from the ET-PD group, and cluster profiles were established through statistical analysis of demographic and clinical variables.Results::The ET-PD group consisted of a high number of patients with a family history of ET exhibiting evident tremor with milder hypokinesia and postural instability symptoms, as compared to the PD group. Through the cluster analysis, two clusters of ET-PD patients were identified. The ET-PD cluster 1 ( n = 34) had a shorter ET duration before PD onset, lower number of patients with a family history of ET, higher unified PD rating scale instability scores, higher non-motor symptoms scores (non-motor symptoms scale D1 scores, Hamilton depression scale scores, Hamilton anxiety scale scores, and PD sleep scale-2 scores), and higher Chinese version of the PD questionnaire-39 scores relative to the ET-PD cluster 2 ( n = 59). Conclusion::ET-PD patients had significantly different characteristics for motor symptoms as compared to PD patients, and may be distinctly divided into two clinical subtypes, namely, the ET-PD complex type and the ET-PD simple type.

Objective To summarize the computed tomography (CT) and magnetic resonance imaging (MRI) features of IgG4-related pancreatitis.Methods The retrospective and descriptive study was conducted.The clinical data of 23 patients with IgG4-associated pancreatitis who were admitted to Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from November 2012 to May 2018 were collected.There were 21 males and 2 females,aged from 45 to 83 years,with an average age of 63 years.Patients underwent upper abdominal enhanced CT and enhanced MRI examinations.Observation indicators:(1) imaging examination of the patients;(2) imaging characteristics on CT and MRI examinations;(3) follow-up.Follow-up using outpatient examination including laboratory and imaging examination was performed to detect clinical symptoms and signs once a month within 3 months postoperatively,once every 3 months within 3 months to 1 year postoperatively,once every 6 months with 1-2 years postoperatively,and once a year after 2 years postoperatively up to August 2018.Measurement data with skewed distribution were represented as M (range).Count data were described as absolute numbers.Results (1) Imaging examination of the patients:19 of 23 patients underwent upper abdominal enhanced CT combined with upper abdominal enhanced MRI scanning,3 underwent upper abdominal enhanced CT combined with upper abdominal MRI plain scanning,and 1 underwent only upper abdominal enhanced MRI scanning.(2) Imaging characteristics on CT and MRI examinations:pancreatic manifestations included 7 aspects.① Location of lesion and morphological classification:of 23 patients,17 were diffuse type,showing sausage-like appearance;4 were focal type,including 2 with mass in the pancreatic head and 2 with mass in the pancreatic body and tail;2 were multiple type,including 1 with mass in the pancreatic head,body and tail,and 1 with mass in the pancreatic body and tail.② Density of lesions on CT examination:of 23 patients,22 received CT plain scanning,including 10 with equal density and 12 with slightly low density;1 didn't receive CT plain scanning.③ Signal of lesions on MRI examination:on T1 weighted imaging,16 of 23 patients showed homogeneous slightly low signal,4 showed isointensity signal,2 showed mixed slightly low signal,and 1 showed slightly high signal.On T2 weighted imaging,21 of 23 patients showed homogeneous slightly high signal,1 showed isointensity signal,and 1 showed mixed slightly high signal.Of 23 patients,19 underwent diffuse weighted imaging (DWI) and 4 didn't undergo DWI.There were 17 patients with slightly high signal and 2 with high signal on DWI.On apparent diffusion coefficient imaging,10 patients showed slightly low signal,and 9 showed low signal intensity.④ Calcification:of 23 patients,2 had multiple calcifications including 1 of diffuse type with calcification located at pancreatic head,and 1 of focal type with calcification located at pancreatic body and tail;20 had no calcification;1 without CT plain scanning cannot be judged calcification.⑤ Enhancement pattern:23 patients showed progressive delayed enhancement of pancreatic lesions on enhancement scanning,homogeneous or heterogeneous enhancement in the arterial phase,and further enhancement in the portal venous phase and delayed phase (enhancement degree of focal type and multiple type was basically consistent with that of normal pancreas).⑥ Halo sign surrounding lesions:of 23 patients,7 had no halo sign,including 2 of diffuse type,3 of focal type,and 2 of multiple type;16 had halo sign (15 of diffuse type and 1 of focal type),including 11 with halo sign surrounding pancreatic body and tail,3 with halo sign surrounding whole pancreas,and 1 with halo sign posterior pancreatic body and tail,and 1 with halo sign anterior pancreatic head.⑦ The main pancreatic duct:14 of 23 patients had irregular stenosis of the main pancreatic duct,7 had mild dilatation of the main pancreatic duct,and 2 had no dilatation of the main pancreatic duct.Extra-pancreatic manifestations included 3 aspects.① Bile duct dilatation:10 of 23 patients had simple intra-hepatic and extra-hepatic bile duct dilatation,8 had no intrahepatic or extra-hepatic bile duct dilatation,and 5 had intra-pancreatic common duct stenosis with upstream bile duct dilatation.② Extra-pancreatic organ involvement:18 of 23 patients had extra-pancreatic organ involvement (5 with gall bladder involvement,4 with intra-hepatic and extra-hepatic bile duct and gall bladder involvement,3 with intra-hepatic and extra-hepatic bile duct involvement,1 with lung involvement,1 with spleen involvement,1 with common duct involvement,1 with kidney involvement,1 with lung and intra-hepatic and extra-hepatic bile duct involvement,1 with fibrosis of mesenteric root),and 5 had no extra-pancreatic organ involvement.③ Retroperitoneal lymph nodes:2 of 23 patients had enlarged retroperitoneal lymph nodes,21 had no enlarged retroperitoneal lymph nodes.(3) Follow-up:of 23 patients,19 were followed up for 3-74 months,with a median time of 22 months.Sixteen of 19 patients had various degrees of improvement in clinical symptoms,laboratory and imaging manifestations after standardized hormonotherapy,and 3 had no response to hormonotherapy.Conclusions CT and MRI examinations have important diagnostic value for IgG4-related pancreatitis.The pancreas mainly show diffuse swelling and halo sign surrounding lesions is a typical imaging characteristic.

OBJECTIVE： To study the correlation of tacrolimus concentrations among transplant patients’ whole blood， plasma and blood cells， analyze the effects of transplant types and ages on the their correlation， and to provide reference for rational drug use in clinic. METHODS： Totally 20 patients receiving tacrolimus anti-rejection therapy after transplantation and undergoing therapeutic drug monitoring （TDM） were randomly selected. According to the type of transplantation， they were divided into renal transplantation group and lung transplantation group （10 cases for each group）.  According to age， they were divided into three groups： 20-40 years old group， 41-60 years old group and 61-80 years old group （4， 9， 7 cases for each group）. Their residual blood after TDM was collected. Chemiluminescence microparticle immuno assay  （CMIA） was used to detect the concentration of tacrolimus in whole blood. UPLC-MS/MS was used to measure the concentrations of tacrolimus in plasma and blood cells. Pairs plots and Spearman rank correlation analysis were used to analyze the correlation of tacrolimus between whole blood and plasma， between whole blood and blood cells， between plasma and blood cells as well as the effects of transplant types and ages on tacrolimus concentrations among tansplant patient’s whole blood， plasma and blood cells. RESULTS： The correlation of tacrolimus concentrations in whole blood and plasma （r＝0.623，P＜0.01） was slightly stronger than that of whole blood and blood cells （r＝0.591， P＜0.01）； while  the correlation of tacrolimus concentration in plasma and blood cells was relatively weak （r＝0.497，P＜0.05）. Transplant type and age had an effect on the correlation of tacrolimus concentrations among patients’ whole blood， plasma， blood cells. The correlation of tacrolimus concen- tration in whole blood， blood cells and plasma in renal transplantation group was also weak （all r＜0.5）， and was weaker than that in lung transplantation group. The correlation of tacrolimus concentration among whole blood， plasma and blood cells was weak in patients of aged 20-40 years old group （all r＜0.3）， and was weaker than that of patients of aged 41-60 years old group and 61-80 years old group. CONCLUSION： Post-transplantation patients’ tacrolimus concentrations in whole blood， plasma and blood cell have a weak correlation. Rejections and adverse effects should be monitored in these patients， especially those renal transplantation patients or those patients aged 20 to 40.

OBJECTIVETo investigate the effect of aluminum exposure on neuronal apoptosis of rats hippocampus and the correlation of and synaptic plasticity.

METHODSThere were 40 SPF grade SD rats which were randomly divided into four groups: the control group, the low dose group, the medium dose group and the high dose group, 10 rats in each group. The rats were daily gavaged with aluminum lactate for 30 days. The hippocampal fEPSPs in rat was measured by electrophysiological grapher and the neuronal apoptosis in hippocampus was detected by Flow cytometer. In addition, the relative expression of gene which includes caspase-3, 8, 9 was measured by Real-time PCR.

RESULTSCompared to the control group, the average of fEPSPs which after HFS 10, 20, 30, 40, 50, 60 min was decreased at different time point in the low dose group, the medium dose group and the high dose group (P < 0.05). Compared with the control group, the rate of apoptosis was significantly increased in the medium dose group and the high dose group (P < 0.05). Compared to the control group, the relative expression of caspase-3 in the medium dose group and the high dose group was significantly increased in Real-time PCR (P < 0.05), and the relative expression of caspase-8 in the high dose group was significantly increased (P < 0.05).

CONCLUSIONAluminum exposure may induced neuronal apoptosis in rats, and then affect hippocampal synaptic plasticity.

Aluminum ; toxicity ; Aluminum Compounds ; toxicity ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Hippocampus ; cytology ; drug effects ; Lactates ; toxicity ; Neuronal Plasticity ; drug effects ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley