Objective To observe the effect of MG132 on the expression of extracellular regulated kinase 1/2 (ERK1/2) and connective tissue growth factor (CTGF) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose (1.5％,2.5％,4.25％) for 24 hours,high glucose (2.5％) for 0,12,24,48 hours,incubated with MG132 (0.5,1,2 μmol/L) for half an hour and then with high glucose (2.5％) for 24 hours.ERK1/2 protein was detected by Western blotting,and CTGF protein in supernatant was detected by ELISA.Results Compared with the control group,the expression of p-ERK1/2 was significantly increased in the groups stimulated by high glucose (P ＜0.01),reached the peak at 24th hour (P ＜ 0.01),and then the expression decreased at 48th hour,but still was higher than that in the normal control group (P ＜ 0.01).CTGF protein expression of RPMCs induced by high glucose increased,in time-and dose-dependent manner (P ＜ 0.05).MG132 could significantly decrease the expression of ERK1/2 and CTGF induced by high glucose (P＜0.05).Conclusions MG132 can decrease the expression of p-ERK1/2 and CTGF in RPMCs induced by high glucose.The ubiquitin proteasome pathway participates in the development of peritoneal fibrosis,and blocking the way may contribute to the prevention of peritoneal fibrosis.

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Applications of network pharmacology are increasingly widespread and methods abound in the field of drug development and pharmacological research. In this study, we choose rosiglitazone compound as the object to predict the targets and to discuss the mechanism based on three kinds of prediction methods of network pharmacology. Comparison of the prediction result has identified that the three kinds of prediction methods had their own characteristics: targets and pathways predicted were not in accordance with each other. However, the calcium signaling pathway could be predicted in the three kinds of methods, which associated with diabetes and cognitive impairment caused by diabetes by bioinformatics analysis. The above conclusion indicates that the calcium signaling pathway is important in signal pathway regulation of rosiglitazone compound, which provides a clue to further explain the mechanism of the compound and also provides a reference for the selection and application of methods of network pharmacology in the actual research.
Calcium Signaling ; Cognitive Dysfunction ; Computational Biology ; Diabetes Mellitus ; Humans ; Pharmacology ; methods ; Thiazolidinediones ; pharmacology

Objective To establish an eukaryotic vector of monkey B virus glycoprotein D gene and analyze the expression of gD gene in human embryonic kidney 293T cells.Method First, the protein of monkey B virus glycoprotein D was obtained by gene synthesis.The gene fragments were digested with Pst I and Not I, and ligated to pEGPF-N3. Then, the recombinant plasmid pEGPF-N3-GD was transfected into 293T cells.The expression of gD protein in the cells was detected by Western blot, and the expression localization was investigated using laser scanning confocal microscopy. Results The recombinant plasmid pEGPF-N3 carrying gD gene was successfully constructed, and normally expressed in the 293T cells.Conclusions Glycoprotein D of monkey B virus is expressed successfully in the 293T cells and the protein is located on the cell surface.It may be useful for the preparation of specific recombinant antigen to the glycoprotein D of monkey B virus on cell surface, and can be also used for preparation of antigen slide for detection of monkey B virus.

Objective To investigate the immunoregulatory effect and compare the different regula- tions of bone marrow mescnchymal stem cells(MSCs)derived from both lupus(NZBWF1/J)and normal(BALB/ c)mice on T lymphocytes in vitro.Methods MSCs from NZBWF1/J and BALB/c mice bone marrow were iso- lated and expanded,and identified by the surface phenotypes.CD3~+ T lymphocytes isolated by nylon wool columns were stimulated by phorbol myristate acetate(PMA)and co-cultured with or without the two strains of MSCs for 24 h.Intracellular eytokines of T cell,such as interferon(IFN)-?,interleukin(IL)-4,IL-12,IL-6, were analyzed by flow cytometry and quantification of transcription factors T-box expressed in T cells(T-bet) and GATA-binding protein 3(GATA-3)were detected by reverse transcriptase PCR(RT-PCR).T cell apop- tosis was assessed by flow cytometry using rhodamine123.Results The results showed that a decrease of CD3~+ T cell apoptosis was seen when NZBWF1/J MSCs or BALB/c MSCs were added to T cells stimulated by PMA(P＜0.05),and an increase of TH2 cytokines by NZBWF1/J MSCs and TH1 eytokines by BALB/c MSCs were observed in the CD3~+ T cells eo-cuhured with MSCs(P＜0.05).Conclusion It is suggested that the al- teration of T subsets caused by MSCs may interfere with the systemic lupus erythematosus(SLE)development and normal MSCs may be effective in the improvement of SLE.NZBWF1/J MSCs have defective immunoregula- tory function when compared with MSCs from healthy mouse strains.

Objective:To compare preoperative planning parameters between non-coplanar and coplanar template-assisted radioactive seed implantation in the treatment of pancreatic cancers, in order to guide clinical application.Methods:Patients with pancreatic cancers who received external irradiation in the Peking University Third Hospital from Jan 2017 to May 2019 were selected.Their image information was imported into the brachytherapy planning system, and the non-coplanar plan and coplanar plan were designed individually.Each patient′s prescription dose was set to 110 Gy, and the activity of the radioactive seeds were 0.4 mCi(1 Ci=3.7×10 10Bq), respectively.For the two plans, the dose distribution was optimized and dosimetric parameters were compared, including the implantation needle number, the implanted seed number, the minimum prescription doses delivered to 90% and 100% of the target volume ( D90 and D100), mean percentages of volume receiving 100%, 150% and 200% of the prescription doses ( V100, V150 and V200), conformity index (CI), external index (EI), and homogeneity index (HI) of the target volume, as well as the doses of 2 cm 3 and 5 cm 3 ( D2 cm 3 and D5 cm 3) of the surrounding normal organs such as the small intestines, colon, duodenum, stomach, and spinal cord. Results:The implantation needle number in the coplanar plan was slightly higher than that in the non-coplanar plan, namely 18.63 vs. 16.45 ( t=-3.239, P <0.05). The implanted seed number was equivalent, namely 90.2 vs. 91.01, with no statistical difference ( P>0.05). There was no significant difference between D90, D100, V100, V150, V200, CI, EI, and HI in the target area of the two plans ( P>0.05). Meanwhile, there was no obvious difference in D2 cm 3 and D5 cm 3 of normal organs including the small intestines, colon, duodenum, stomach, and spinal cord ( P>0.05). Conclusions:With both the coplanar plan and the non-coplanar plan, the prescription doses can be achieved and meanwhile, there are very small differences in the doses of normal organs.Given that 3D-printing non-coplanar and coplanar templates have their own characteristics, it is necessary to choose them according to specific situations.

Objective To study the correlation between human papillomavirus(HPV) infection and cervical immune function.Methods A total of 209 women with HPV positive results in HPV-infected group,40 women with HPV negative results in control group,cervical samples were detected and genotyped with HPV GenoArray Diagnostic Kit.Concentrations of cytokines including interleukin(IL)-2,IL-4,interferon(IFN)-γ,and transforming growth factor(TGF)-β in cervical specimens were measured by ELISA.Results IL-2,IL-4 and IFN-γ levels in the HPV-infected group were different with those of the control group(P=0.000 1,0.001 0,0.000 1),TGF-β in HPV-infected group and control group showed no significant difference(P=0.680 0).IFN-γ in high-risk HPV group was significant lower than that in the low-risk HPV group(P=0.007 0).Conclusion HPV infection might be one of the main reasons of cervical local immune dysfunction.

Objective To observe the effect of silibinin on the expression of integrin linked kinase (ILK),transforming growth factor β1 (TGF-β1) and α-smooth muscle actin (α-SMA) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose(1.5％,2.5％,4.25％)for 24 hours,high glucose (2.5％) for 12,24,48,72 hours,high glucose (2.5％) for 24 hours after silibinin (5,10,20 mg/L) preincubate for 2 hours.ILK and α-SMA mRNA were detected by real-time PCR.ILK protein was detected by Western blotting.TGF-β1 protein in supernatants was detected by ELISA.Results Compared with the control group,the expresssion of ILK,TGF-β1 and α-SAM was significantly increased in groups stimulated by high glucose (all P ＜ 0.05).Silibinin could significantly decrease the expression of ILK,TGF-β1 and α-SMA induced by high glucose (all P ＜ 0.05).Conclusions High glucose can up-regulate the expression of ILK,TGF-β1 and α-SMA.Silibinin can reverse these changes.

ObjectiveTo evaluate the effects of cardiac resynchronization therapy (CRT) on improving left ventricle (LV) function and LV asynchrony in patients with dilated cardiomyopathy(DCM)using three-dimensional ultrasound speckle tracking imaging(3D-STI).MethodsTwenty one patients with DCM and 25 normal subjects were enrolled in this study.Parameters including LV mean longitudinal peak strain (MLS),LV mean circumferential peak strain (MCS),LV mean radial peak strain (MRS),LV ejection fraction(3D-LVEF),maximal temporal difference in area strain of LV 16 segments (A-MaxTs) and its standard deviation (A-Ts-SD) were investigated by 3D-STI.LV ejection fraction (ECT-LVEF) and LV phase angle width (LVW) were obtained by radionuclide ventriculography and phase image analysis.Results The indexes of MLS,MCS,MRS and 3D-LVEF were decreased and A-MaxTs,A-Ts-SD were prolonged in DCM group compared with those of control group.There was a slightly improvement in MRS and ECTLVEF 1 week after CRT,but other Parameters including MLS,MCS,A-MaxTs,A-Ts-SD and 3D-LVEF had no significant change.Compared with corresponding values before and 1 week after CRT,all parameters (MCS,MRS,A-MaxTs,A-Ts-SD,3D-LVEF and ECT-LVEF) were further improved 6 months after CRT.MLS,MCS,MRS and 3D-LVEF had good relationship with ECT-LVEF,and A-MaxTs,A-Ts-SD had good relationship with LVW in DCM group.Conclusions3D-STI is a simple and accurate method for evaluating effects of CRT on improving LV function and LV asynchrony in patients with DCM.

Objective To investigate the relationship between expression of Her-2 gene and the transfer number of axillary lymph node and its influence on prognosis.Methods A total of 351 cases with primary breast cancer from January 2008 to January 2011 were selected.The expression of Her-2 gene was detected by immunohistochemical method,and analyzed the relationship between it and the transfer number of axillary lymph node and its influence on prognosis.Results The expression of Her-2-,+,++ had no correlation with the transfer number of axillary lymph node (x2 =3.757,1.650,2.379,P ＞ 0.05),while the expression of Her-2 +++ had correlation with the transfer number of axillary lymph node (x2 =8.681,P ＜ 0.05).The 2 years survival rates in the expression of Her-2-,+,++,+++ were 77.01％ (201/261),85.00％ (34/40),29.17％(7/24),1 1.54％ (3/26),and which in the expression of Her-2--+ was significantly higher than that in the expression of Her-2 ++-+++ (x2 =61.605,P ＜ 0.01).The transfer number of axillary lymph node was 0 and 1-3,the 2 years survival rate in the expression of Her-2--+ was significantly higher than that in the expression of Her-2 ++-+++,and there was significant difference (x2 =61.605,14.747,P ＜ 0.05).The transfer number of axillary lymph node was 4-9 and ≥ 10,there was no significant difference in the 2 years survival rate between the expression of Her-2--+ and Her-2 ++-+++ (x2 =3.691,3.482,P ＞ 0.05).Conclusions The expression of Her-2-,+,++ has no correlation with axillary lymph node metastasis,and the 2 years survival rate in the expression of Her-2--+ is higher than that in the expression of Her-2 ++-+++,while Her-2-has no difference with Her-2 + in prognosis.While the transfer number of axillary lymph node ≤ 3,the expression of Her-2 gene may be an important prognostic factor.