Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.

Objective To compare the difference of the HBsAg detected results between the Roche cobas e601 electrochemilumi-nescence immunoassay instrument and the Abbott Architect i2000chemiluminescent microparticle immunoassay instrument.Methods The HBsAg positive specimens with the quantitation results of lower than 250IU/mL detected by the Abbott Architect i2000 were selected and simultaneously detected by the Roche cobas e601.The differences of detected results were compared and per-formed the linear correlation and analysis regression.Results 46 clinical specimens were detected.The detected results had best correlation between the two instruments by getting rid of 1 specimen with unconformable reactivity of detected results.15 speci-mens had the HBsAg detected result of 0.05-1.00 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =17.49X+0.843(r=0.979);15 specimens had the HBsAg detected result of 1.1 -10.00 IU/mL IU/mL by the Abbott Architect i2000,the linear regression equation was Y =15.72X +21.06(r=0.952);15 specimens had the HBsAg detected result of 11 -250 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =29.17X -129(r=1.000).Conclusion The detected results have better correlation between the two instruments and can be mutually converted by the formulas.

AIM:To study the effect of Hand2 (one of basic helix-loop-helix proteins’ transcription factors) expression on the development of the cardiac tissues in the fetal rats from gestational diabetes mellitus ( GDM) parents, and to investigate the potential pathogenesis of GDM-induced congenital cardiac defects in rats.METHODS: The adult Spra-gue-Dawley female rats were randomly divided into blank control group (n=24), GDM group (n=30), negative control group ( n=30) and insulin intervened group ( n=30) .The GDM model was established by intraperitoneal injection of 2%streptozotocin (STZ, 40 mg/kg body weight) to the pregnant rats on the successive day.The rats in insulin intervened group were injected with intermediate-acting insulin in order to keep the fasting blood glucose in the normal range.The rats in negative control group were injected with citric acid-sodium citrate buffer solution in the same position.Blood glucose and body weight were examined every day 72 h after STZ injection.On E12, E15 and E19, the rats were anesthetized and the embryonic cardiac tissues were collected after caesarean section.The histopathological changes of the cardiac tissues were observed under microscope with HE staining.The expression of Hand2 was analyzed by the method of immunohisto-chemistry.The expression of Hand2 in the cardiac tissues at mRNA and protein levels was determined by real-time fluores-cence quantitative PCR and Western blotting.RESULTS:During the development of embryonic heart, the protein expres-sion of Hand2 in the cardiac tissues was showed dynamic changes.Observed on E12, obviously increased on E15, and at the highest level on E19.Compared with the other 3 groups, the protein and mRNA expression of Hand2 in GDM group was decreased at the time points of E12 and E15.CONCLUSION:The morbidity of fetal cardiac malformation is significantly increased in GDM group, suggesting that Hand2 may be involved in the development of cardiac malformation in GDM.

group was significantly longer than that in the low MELD score group (P <0.05).And there was no significant difference in the length of hospital stay,mortality during hospitalization,incidence of early complications, follow-up time and overall survival rate between two groups (all in P >0.05).The peak level of aspartate aminotransferase (AST)in the high MELD score group was significantly higher compared with that in the low MELD score group (P <0.05 ).The incidence of bile leakage,abdominal abscess and liver dysfunction significantly differed between two groups (all in P <0.05 ).Conclusions It is a safe and short-term efficacious approach to perform liver transplantation with organs obtained from Chinese donation after citizens’ death in patients with high MELD score liver recipients.

[摘 要] 目的：探讨β-1,6 N-乙酰氨基葡萄糖转移酶2（GCNT2）基因在胃癌（GC）组织中的表达及其在GC发生、发展和诊断及预后中的作用。方法：利用TIMER、GEPIA2、Oncomine和UALCAN等数据库数据，以及2018年1月至2019年12月滨州医学院附属医院手术切除的25例GC患者的癌和配对癌旁组织标本，分析GCNT2基因在GC组织中的表达及其在GC诊断和预后中的价值，利用LinkedOmics、GSEA和ssGSEA分析GCNT2所涉及的主要信号通路及其与免疫浸润之间的相关性。将pc-GCNT2及其阴性对照质粒转染进胃癌SGC-7901和BGC-823细胞，用克隆形成实验和Transwell实验检测GCNT2对GC细胞增殖和侵袭的影响，WB法检测细胞中GCNT2、STAT3和PD-L1蛋白的表达水平。结果：GCNT2 mRNA在GC组织中的表达水平显著低于癌旁组织（P<0.05或P<0.01），其表达水平与患者预后显著相关（P<0.05），其对GC诊断有较高的价值。GCNT2在GC组织中的甲基化状态显著高于癌旁组织，GCNT2基因参与的生物过程主要是参与细胞形态发生的成分、细胞间黏附、多细胞生物信号和突触传递等。单基因GSEA分析发现，GCNT2在GC中主要抑制IL-6/JAK/STAT3、IL-2/STAT5信号通路和炎症反应、α/γ干扰素响应与NF-κB表达等。GCNT2的表达与GC组织的免疫浸润具有显著相关性。过表达GCNT2可显著抑制GC细胞的增殖和侵袭能力（均P<0.01），降低细胞中STAT3和PD-L1的表达水平（均P<0.01）。结论：GCNT2基因在GC组织中低表达，与GC的诊断及预后显著相关，其主要通过抑制IL-6/JAK/STAT3和免疫相关致癌信号通路而在GC的发生、发展中发挥重要的作用。

The eutR gene deletion mutant of Salmonella enteritidis was successfully constructed by homologous recombination.Through the study of its biochemical characteristics,motility,resist-ance to stress in vitro and survival ability in RAW 264.7 cells,it was found that the biochemical characteristics and motility of the eutR gene deletion mutant of Salmonella enteritidis had no sig-nificant change compared with the wild type of Salmonella enteritidis.The ability of eutR gene de-letion strain of Salmonella enteritidis to resist acid,alkali and oxidation was significantly reduced,while the ability to resist heat was not significantly changed;the survival ability of eutR gene dele-tion strain in RAW 264.7 cells was significantly reduced compared with the wild type.In order to further analyze the effect of eutR gene on the expression of virulence factors of Salmonella enterit-idis,the relative expression levels of invH,ssav,ssrA,xthA,orf245,sodC,lrp,mrr1 and hflk virulence genes of the deletion strain and the wild strain were detected by SYBR Green PCR.It was found that the expression of the virulence factors mentioned above in the eutR gene deletion strain of Salmonella enteritidis was significantly down-regulated compared with that in the wild-type strain.The LD50 of the eutR gene-deleted strain of Salmonella enteritidis was determined by ani-mal experiments,and the results showed that the LD50 of the eutR gene-deleted strain was higher than that of the wild-type strain,indicating that the eutR gene could affect the virulence of Salmonella.This study clarified the effect of eutR gene on the survival ability,some biological characteristics and virulence of Salmonella enteritidis in macrophages,and provided a new gene knockout target for the development of attenuated Salmonella enteritidis genetic engineering vac-cine.

AIM:To investigate the effects of Balanophora involucrata Hook.f.(BIH)extracts on bile acid metabolism and liver injury in mice with metabolic dysfunction-associated fatty liver disease(MAFLD)through the gut mi-crobiota-farnesoid X receptor(FXR)axis,and to explore the underlying mechanisms.METHODS:Forty C57BL mice were randomly divided into control group,MAFLD model group,medium-dose BIH group,and high-dose BIH group.The mice in control group received a regular diet,while those in other groups were fed with high-fat diet for 12 weeks to induce MAFLD.The mice in medium-and high-dose BIH groups received 0.598 and 0.299 g/kg BIH solution,respectively,while those in control and MAFLD groups received an equivalent volume of normal saline.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were measured using an automatic biochemical analyzer.Liver morphology,steatosis and fibrosis were assessed by HE,oil red O and Masson staining.Levels of TC,tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in liver tissues,and bile acids in serum and ileum tissues were measured by ELISA.Protein expression of FXR and fibroblast growth factor 15(FGF15)in ileum tissues,and FXR,small heterodimer partner(SHP)and cholesterol 7α-hydroxylase(CYP7A1)in liver tissues were analyzed by Western blot.Intestinal microbiota changes were assessed by 16S rRNA gene sequencing.RESULTS:(1)The MAFLD mice exhibited increased serum TC,TG,LDL-C and bile acid levels,liver TC,TNF-α and IL-6 levels,and lipid deposition.However,BIH intervention improved these factors and increased FXR and SHP pro-teins,but decreased CYP7A1 expression in the liver.The protein levels FXR and FGF15 in the ileum were also elevated.(2)Intestinal flora analysis demonstrated that BIH intervention improved the abundance and diversity of intestinal flora in MAFLD mice.Specifically,there was an increase in Bacteroidetes/Firmicutes ratio and a decrease in Proteobacteria and Verrucomicrobia.At the genus level,abundance of Duncaniella,Muribaculum and Paramuribaculum increased,while He-licobacter decreased.CONCLUSION:Treatment with BIH regulates intestinal flora,decreases FXR levels,enhances CYP7A1 expression,promotes bile acid synthesis,reduces hepatic cholesterol accumulation,and attenuates liver steato-sis and inflammation in MAFLD mice,indicating potential therapeutic effects.

Objective: To evaluate the endothelial function in obstructive sleep apnea syndrome(OSAS) children and to identify related factors of endothelial dysfunction. Methods: This was a cross-sectional study. Children with habitual snoring (snoring ≥3 nights per week) admitted to the ward of otolaryngology, head and neck surgery, Beijing Children′s Hospital were recruited to this study between 1^st June 2015 and 1^st March 2016. All children aged 3 to 11 years and of them 245 were boys and 110 were girls. All subjects underwent an overnight polysomnography (PSG), as well as endothelial function testing. All subjects were grouped into primary snoring (PS) and OSAS group according to the obstructive apnea hypopnea index (OAHI). T test or Wilcoxon test were used to compare the differences in PSG results between the two groups, and univariate and multivariate correlation analyses were used to explore the relevant factors affecting the endothelial function. Results: A total of 355 subjects were enrolled and 248 had OSAS, and 107 had PS. There were no significant differences in age, gender and body mass index (BMI) Z-score between the two groups (all P>0.05). OSAS group had higher OAHI, oxgen desaturation index and respiratory related arousal index (5.2 (2.2, 13.2) vs. 0.4 (0.1, 0.7), 4.1 (2.0, 13.1) vs. 0.5 (0.1, 1.0), 2.5 (1.0, 4.8) vs. 0.4 (0.1, 0.9), Z=-14.957, -11.790, -10.378, all P<0.01), and lower minimum oxygen saturation and reactive hyperemia index (RHI) than those of PS (0.89 (0.85, 0.92) vs. 0.94 (0.91, 0.95), 1.2±0.2 vs. 1.1±0.1, Z=-9.337, t=5.354, P<0.01). Univariate regression analysis showed that RHI was linearly correlated with age (parameter estimate=0.017, P<0.01), gender (parameter estimate=0.065, P<0.01), OAHI (parameter estimate=-0.023, P<0.01), oxygen desaturation index (parameter estimate=-0.019, P<0.01), respiratory related arousal index (parameter estimate=-0.031, P<0.01), and oxygen saturation nadir (parameter estimate=0.067, P=0.045). The relationship between BMI Z-score and RHI was quadratic. Multivariate regression analysis showed that age (parameter estimate=0.015, P<0.01), BMI Z-score (parameter estimate=0.040, P<0.01), BMI Z-score quadratic form (parameter estimate=-0.010, P<0.01), respiratory related arousal index (parameter estimate=-0.020, P<0.01) were independently correlated with RHI. Conclusions Children with OSAS have significant endothelial dysfunction compared with PS. Frequent arousals due to obstructive respiratory events during sleep may be a candidate risk factor for endothelial dysfunction in children with OSAS.

To investigate the γ pass rate limit of plan verification equipment for volumetric modulated arc therapy (VMAT) plan verification and its sensitivity on the opening and closing errors of multi-leaf collimator (MLC), 50 cases of nasopharyngeal carcinoma VMAT plan with clockwise and counterclockwise full arcs were randomly selected. Eight kinds of MLC opening and closing errors were introduced in 10 cases of them, and 80 plans with errors were generated. Firstly, the plan verification was conducted in the form of field-by-field measurement and true composite measurement. The γ analysis with the criteria of 3% dose difference, distance to agreement of 2 mm, 10% dose threshold, and absolute dose global normalized conditions were performed for these fields. Then gradient analysis was used to investigate the sensitivity of field-by-field measurement and true composite measurement on MLC opening and closing errors, and the receiver operating characteristic curve (ROC) was used to investigate the optimal threshold of γ pass rate for identifying errors. Tolerance limits and action limits for γ pass rates were calculated using statistical process control (SPC) method for another 40 cases. The error identification ability using the tolerance limit calculated by SPC method and the universal tolerance limit (95%) were compared with using the optimal threshold of ROC. The results show that for the true composite measurement, the clockwise arc and the counterclockwise arc, the descent gradients of the γ passing rate with per millimeter MLC opening error are 10.61%, 7.62% and 6.66%, respectively, and the descent gradients with per millimeter MLC closing error are 9.75%, 7.36% and 6.37%, respectively. The optimal thresholds obtained by the ROC method are 99.35%, 97.95% and 98.25%, respectively, and the tolerance limits obtained by the SPC method are 98.98%, 97.74% and 98.62%, respectively. The tolerance limit calculated by SPC method is close to the optimal threshold of ROC, both of which could identify all errors of ±2 mm, while the universal tolerance limit can only partially identify them, indicating that the universal tolerance limit is not sensitive on some large errors. Therefore, considering the factors such as ease of use and accuracy, it is suggested to use the true composite measurement in clinical practice, and to formulate tolerance limits and action limits suitable for the actual process of the institution based on the SPC method. In conclusion, it is expected that the results of this study can provide some references for institutions to optimize the radiotherapy plan verification process, set appropriate pass rate limit, and promote the standardization of plan verification.
Humans ; Radiotherapy, Intensity-Modulated ; Immune Tolerance ; Nasopharyngeal Carcinoma ; ROC Curve ; Nasopharyngeal Neoplasms/radiotherapy*