Objective To investigate the effect of CKLF-like MARVEL transmembrane domain contai-ning member 5(CMTM 5)on EG-VEGF expression in prostate cancer cells , and to detect the potential mechanism of CMTM5 inhibiting prostate cancer .Methods The relative expression of CMTM 5 and EG-VEGF in prostate cells and prostate cancer cells was detected .Prostate cancer cells were given Plasmid transfection to overexpress CMTM5 and EG-VEGF expression was again detected .Then CMTM5 and EG-VEGF were compared between be-fore and after CMTM5 plasmid transfecting prostate cancer cells .Results Compared with the relative expresion of EG-VEGF and CMTM5 in prostate cells , prostate cancer cells showed high expression of EG-VEGF and low ex-pression of CMTM5, which were statistically significant , P<0.05.Compared with prostate cancer cells , the rela-tive expression of CMTM5 were obviously upregulated and EG-VEGF obviously decreased in prostate cancer cell after transfected by CMTM5 plasmid, which were statistically significant , P<0.05.Conclusions Prostate canc-er cells shows higher EG-VEGF expression and lower CMTM 5 campared with normal prostate cells .EG-VEGF is suppressed significantly when the prostate cancer cell is transfected by CMTM 5 plasmid and shows highlevel of CMTM5 expression , suggesting high expression of CMTM 5 may inhibit the development of prostate cancer by downregulating EG-VEGF expression .

Objective To establish a HPLC method for the determination of Shionone in Ziwan Zhisou granule. Methods Phenomenex C18 column (250? 4.6 mm) was used. The mobile phase was acetonitrile and the detection wave length was at 203 nm. Result A good linearity of Shionone was in the range of 0.025～ 0.250 ? g/? L, r=0.9996. The average recovery was 97.8 % , RSD was 2.37 % ( n=5) . Conclusion This method is sensitive, accurate and simple and with good reproducibility for the determination of Shionone in Ziwan Zhisou granule. 

Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25 T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the therapy of type 1 diabetes.Methods CD4+ CD25-T cells of 10 normal people and 10 newly-onset type 1 diabetic patients were separated from peripheral blood by MACS immunomagnetic beads and stimulated by Human T-Activator CD3/CD28 Dynabeads to proliferate.CFSE labeling technique was used to evaluate the proliferation of CD4+ CD25-T cells by flow cytometry.Liraglutide of different concentrations(0,25,50,and 100 nmol/ml) was added to the proliferation system,then the proliferation of CD4+CD25-T cell was measured.Results (1) Liraglutide suppressed the proliferation of CD4+ CD25-T cells from either normal people or type 1 diahetic patients with dose-dependent manner (P ＜ 0.05).(2) Under the different concentrationsofliraglutide,the proliferation ofCD4+CD25 T cells from diabetic patients was mueh more robust than that of normal people (P＜0.01).(3) The inhibitory effects of liraglutide on CD4+ CD25-T cells proliferation in normal people and diabetic patients were similar (P＞0.05).Conclusion The proliferation of CD4+ CD25 T cells in type 1 diabetic patients was more robust than normal people,which indicated cellular immune dysfunction in type 1diabetes.Liraglutide inhibits the proliferation of CD4+ CD25-T cells of type 1 diabetic patients in vitro.The immunosuppression effect of liraglutide may have potential value in the treatment of type 1 diabetes.

Multiple Linear Regression (MLR) is a generalization of simple linear regression and is one of the commonly used models in multivariate statistical analysis. This article introduces the MLR model from the perspective of practical application. Four parts, including basic principle, application examples, the application condition and diagnosis, and the extension of the model, are sequentially illustrated in this article. Particularly, in the last part, alternative methods of the model are introduced when the application condition of the model is not met. We sincerely hope that this article could make our audiences have a better understanding of the MLR model in order to improve the efficiency of data utilization and statistical analysis by correctly performing this model in their research.

Analyses of the multivariate regression model are ued very widely in the medical research. Analytical methods of the mutivariate regression model including multiple linear regression, logistic regression, Poisson regression and Cox proportional hazard model were introduced in this article. The contents of the article covered the application conditions of regression models, analytical procedures, strategies of selecting independent variables, extended discussions of regression models and application notes. It is expected that authors could understand the principle of the mutivariate regression model, accurately use these analytical methods in their research, improve the efficiency of data utilization, and enhance the level of statistical analyses.

Logistic regression is a kind of multiple regression method to analyze the relationship between a binary outcome or categorical outcome and multiple influencing factors, including multiple logistic regression, conditional logistic regression, polytomous logistic regression, ordinal logistic regression and adjacent categorical logistic regression. This paper illustrates the basic principle, independent variable selection and assignment, applied condition, model evaluation and diagnosis for multiple logistic regression model. Moreover, the principle and application for polytomous logistic regression and ordinal logistic regression models were also introduced. By providing SAS codes and detailed explanations of the result for an example of obesity, readers could be able to better understand logistic regression model, and apply this method correctly to their research and daily work, so as to improve their capacity of the data analysis.

Objective To evaluate the imaging features of nigrosome-1 in Parkinson's disease (PD) with a 3 T scanner by susceptibility weighted imaging (SWI),and to explore its clinical relevance.Methods Thirty-two patients with primary PD diagnosed by neurologists were collected.Healthy controls matched to their age and gender were recruited during the same period (n=20).All subjects underwent routine brain magnetic resonance imaging (MRI) and sensitive weighted imaging (SWI).The SWI images of the subjects were evaluated to evaluate nigrosome-1 by blinded investigators.Then,the correlation between imaging features and clinical data was analyzed.Results In the PD group,21 cases of bilateral "absent swallow-tail sign",five cases of bilateral "indecisive swallow-tail sign",five cases of "absent swallow-tail sign" on one side and "indecisive swallow-tail sign" on the other side,and one case of bilateral "clear swallow-tail sign" were found.The course of the "absent swallow-tail sign" group (56 (54) months) was significantly longer than the "non-absent swallow-tail sign" group (18 (18) months;U=-2.47,P=0.01).The Hoehn-Yahr stage was significantly higher in the "absent swallow-tail sign" group (2.0 (0.5)) than in the "non-absent swallow-tail sign" group (1.5 (0.5),U=-2.21,P=0.03).There was also a statistically significant difference in the Unified Parkinson's Disease Rating Scale score (24 (8),13 (14)) between the two groups (U=-2.91,P=0.01).However,there were no statistically significant differences between the two groups in the Hamilton Depression Scale score (5 (2) vs 5 (7),U=-0.10,P=0.94) and the Hamilton Anxiety Scale score (3.0 (2.5) vs 3.0 (3.0),U=-0.02,P=1.00).Conclusion The images of nigrosome-1 by SWI are closely related to the severity of the condition and motor symptoms of patients with PD,which can reflect the severity of the disease.

Objective:To evaluate the feasibility of a new ultrasonic parameter to assess right ventricular-pulmonary artery (RV-PA) coupling in patients with acute pulmonary embolism (APE).Methods:A retrospective analysis was performed in 140 patients with APE diagnosed by computed tomography pulmonary angiography (CTPA) in the Second Affiliated Hospital of Harbin Medical University from August 2017 to June 2020. According to the tricuspid annular plane systolic excursion/pulmonary arterial systolic pressure (TAPSE/PASP) ratio cutoff value 0.40 mm/mmHg reported by the European Society of Cardiology in 2020, the patients were divided into the coupling group ( n=99) and the uncoupling group ( n=41). The conventional ultrasonic parameters of the 2 groups were measured, and then several ultrasonic parameter ratios were obtained. The new ultrasonic parameter, which can replace the TAPSE/PASP ratio, was screened out by Spearman correlation analysis, and ROC curve was plotted to calculate the diagnostic efficacy of this parameter. Results:①Compared with the coupling group, patients in the uncoupling group were older and more likely to be accompanied by dyspnea and venous thrombosis in the lower extremities (all P<0.05), but there was no significant difference in other general data(all P>0.05); ②Compared with the coupling group, tricuspid regurgitation velocity (TRV), tricuspid regurgitation pressure gradient(TRPG), PASP, right ventricle end-diastolic transverse diameter(RVTD), inferior vena cava(IVC) diameter and the ratio of early diastolic tricuspid inflow to tricuspid lateral annular velocity(E/e′), in the uncoupling group increased significantly (all P<0.05), and TAPSE, peak systolic velocity of tricuspid annulus(s′), TAPSE/PASP ratio, TAPSE/TRPG ratio, TAPSE/RVTD ratio and s′/TRPG ratio decreased significantly (all P<0.05); ③The TAPSE/TRPG ratio was highly correlated with TAPSE/PASP ratio ( rs=0.970, P<0.001); The TAPSE/TRPG ratio was still highly correlated with TAPSE/PASP ratio in the uncoupling and coupling groups ( rs=0.966, 0.922; all P<0.001). ④ROC analysis showed that the area under curve for TAPSE/TRPG in diagnosing RV-PA coupling was 0.992. At the cutoff of TAPSE/TRPG <0.625 mm/mmHg for indicating RV-PA coupling, the sensitivity and specificity were 97.6% and 92.9%, respectively. Conclusions:TAPSE/TRPG ratio can be used as a new ultrasonic parameter to reflect RV-PA coupling, which is helpful for clinical identification of APE patients with high risk and poor prognosis.

OBJECTIVE: To establish a method for in vitro expansion of human natural CD4⁺CD25⁺ T regulatory cell (Treg) cells for clinical study and immunotherapy. METHODS: Human natural CD4⁺CD25⁺ Treg were isolated from peripheral blood monocyte cells (PBMCs) by magnetic activated cell sorting (MACS) and expanded by CD3/CD28 expansion beads, IL-2 and rapamycin. The number and the viability of the freshly isolated and expanded Treg were detemined by trypan blue staining. The phenotype and the purity of the freshly isolated and expanded Treg were analyzed by FACS. Treg suppression activity was assessed by mixed lymphocyte reaction (MLR) assay. RESULTS: Human natural Treg were expanded up to 2 000 folds after 3 weeks in culture, and the activity was more than 97%. The expanded Treg retained Treg phenotype as shown by their freshly isolated counterparts, and the purity of CD4⁺CD25⁺FoxP3⁺ Treg was (94.22 ± 2.12)%. The expanded Treg demonstrated a similar potent suppression of both proliferating auto- and allo- CD4⁺CD25⁻ effector T cells in vitro in a cell number-dependent manner. CONCLUSION An in vitro expansion of human natural Treg was established to obtain large numbers of human Treg with highly suppressive phenotype and function, thereby providing a solution to the availability of sufficient human natural Treg in clinical study and immunotherapy.
Cell Culture Techniques ; Cell Separation ; Cells, Cultured ; Humans ; Interleukin-2 ; Leukocytes, Mononuclear ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes, Regulatory ; cytology