Objective To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.Untransfected HK-2 cells and those transfected with empty vector were used as controls.The TLR4 expression was detected by real-time PCR and Western blotting.The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry,and the contents of IL-4 and IFN-γ in the supernatant were detected by EIISA.Results Compared with control groups,the number of apoptotic cells was significantly increased in the HBX transfection group (P ＜ 0.05),and the expressions of TLR4,MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05).IFN-γ level in the supernatant of HBX transfection group was higher (P ＜ 0.05),but IL-4 level was lower as compared to control groups (all P ＜ 0.05).Conclusions Over-expression of HBX gene may induce apoptosis of HK-2 cells and upregulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.

Objective To investigate the effects of hepatitis B virus X (HBX) gene on cell morphology and transdifferentiation of human proximal tubular epithelial cells.Methods The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation.The expression of HBX was confirmed by Q-PCR and Western blotting.Untransfected HK-2 cells and those transfected with empty vector were used as control.The morphology of HK-2 cells was observed by microscopy,the expressions of differentiation marker proteins α-SMA and E-cadherin were detected by Western blotting and Q-PCR,and the contents of IL-1,IL-6 and TNF-α in the supernatant were detected by ELISA assay.Results HBX was successfully expressed in HK-2 cells after transfection.After transfection of HBX gene,the shape of HK-2 cells became irregular,HK-2 cells significantly expressed E-cadherin and α-SMA,and had high levels of IL-1,IL-6 and TNF-α in the cell supernatant (P＜0.01).Conclusion Overexpression of HBX gene in renal tubular epithelial cells may damage cell morphology and promote the occurrence of epithelial-mesenchymal transdifferentiation,which may be related to the inflammatory microenvironment.

Astrocytoma ; blood supply ; pathology ; Capillaries ; pathology ; Central Nervous System Neoplasms ; blood supply ; pathology ; Diagnosis, Differential ; Ependymoma ; blood supply ; pathology ; Glioblastoma ; blood supply ; pathology ; Hemangioblastoma ; blood supply ; pathology ; Humans ; Hyperplasia ; Microvessels ; pathology ; Neoplasms, Neuroepithelial ; blood supply ; pathology ; Oligodendroglioma ; blood supply ; pathology ; Paraganglioma ; blood supply ; pathology

Objective To investigate the effects of different doses of propofor on ATP-sensitive K~+currents(I_KATP)in human atrial myocytes and the underlying mechsnism.Methods A small piece of myocardiumwas obtained from right atrium in patients undergoing atrial septal defect or ventricular septal defect surgery.Themyocardium specimen was placed in cold Ca~(2+)-free cardioplegic solution aerated with 100% oxygen.Themyocardium specimen was cut into small chunks(1 mm~3).Atrial myocytes were isolated by enzymatic dissociationtechnique.The effects of propofol on I_KATP in atrial myocytes were studied using the whole-cell configuration ofpatch-clamp technique.Results The outward currents were recorded with a pipitte solution containing 0.3mmol?L~(-1) ATP.The currents were inhibited by glibendamide 10 ?mol?L~(-1),a specific K_ATP channel inhibitor,suggesting that the outward currents were I_KATP.I_KATP aws activited by propofol in a dose-dependent manner.Conclusion Propefol can activate the I_KATP in human myocytes in a concentration-dependent manner and themechanism of its myocardial depressant action may be partly explained.

Objective To explore prevention and cure effects of the freeze-dried human fibrin glue as the way of the lacrimal duct embolization on xerophthalmia in perimenopausal female rabbit.Methods A total of 72 female rabbits,after anti infection treatment and were cut off third eyelid,were made into perimenopausal xerophthalmia rabbit models.After surgery,all of these rabbits were randomly divided into 6 groups (12 rabbits per group):No treatment group after surgery (group A),PBS prevention group (group B),freeze-dried human fibrin prevention group (group C);no treatment group after modeling (modeling time:Two weeks after surgery,group D),PBS treatment group (group E),freeze-dried human fibrin treatmentgroup (group F).The Schirmer test (SIT),corneal fluorescein (FL) and corneal confocal microscope were performed before and 2 weeks,4 weeks,6 weeks after injection.Results There were statistical differences in FL score and SIT in group A,group B and group C among different time points (F =27.346,10.608;P =0.000,0.001);There were statistical differences between FL scores and SIT among three groups (F =7.579,6.786;P =0.002,0.007);There was significant difference in FL scores and SIT trends among three groups(F =44.897,3.424;P =0.000,0.045).The FL score and SIT of group D,group E and group F were significantly improved after treatment for 2 weeks,4 weeks and 6 weeks,the differences were statistically significant (t =2.906,3.654,4.504;P =0.022,0.017,0.013.t =4.573,5.759,7.231;P =0.032,0.019,0.008);The difference between FL score and SIT in group E and group F was statistically significant after treatment (t =2.776,4.124,5.324;P =0.032,0.026,0.017.t =1.969,3.122,4.324;P =0.038,0.023,0.009).After injection of 6 weeks,the epithelial basal cells (F =17.306,P =0.002) and inflammatory cells (F =34.024,P =0.000) of group A,B and C were significant changed,the differences were statistically significant.After injection of 6 weeks,the epithelial basal cells (F =3.749,P =0.042)and inflarnmatory cells(F=8.806,P =0.005) of group D,E and F were significant changed,the differences were also statistically significant.Conclusion Lacrimal duct embolization with freeze-dried human fibrin glue is effective for the xerophthalmia.

Objective:To explore the monitoring value of left ventricular functional parameters obtained by bedside ultrasound combined with clinically relevant indicators in patients with veno-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A retrospective study was conducted. A total of 24 patients receiving VA-ECMO adjuvant support in Renmin Hospital of Wuhan University from June 2018 to January 2020 were selected. The bedside ultrasound was performed on the first day of ECMO support, the day before weaning, the clinical indicators before weaning were obtained. The differences in clinical indicators and the left ventricular functional parameters between the two groups of whether weaning successfully were compared; univariate Logistic regression analysis was used to screen out the related factors affecting weaning.Results:Sixteen patients were successful weaned and 8 patients failed. Compared with the weaning failure group, patients in the weaning success group required less continuous renal replacement therapy (CRRT, cases: 4 vs. 6, P < 0.05), mean arterial pressure (MAP) before weaning was higher [mmHg (1 mmHg = 0.133 kPa): 84.64±9.55 vs. 62.30±8.79, P < 0.05], and the pulse oxygen saturation (SpO 2) was also higher (0.966±0.670 vs. 0.866±0.061, P < 0.05), while vasoactive-inotropic score (VIS), serum creatinine (SCr) and serum lactic acid (Lac) were lower [VIS score: 7.27±1.42 vs. 16.93±8.52, SCr (μmol/L): 123.60±83.64 vs. 213.10±117.39, Lac (mmol/L): 1.94±0.91 vs. 5.62±5.48, all P < 0.05]. Univariate Logistic regression analysis showed that the MAP, VIS, SCr, Lac, SpO 2 before weaning were the related factors affecting weaning [odds ratio ( OR) were 0.306, -0.740, -0.011, -0.632, -4.069; 95% confidence interval (95% CI) were 1.065-1.732, 0.235-0.899, 0.979-0.999, 0.285-0.992 and 0.001-0.208; P values were 0.014, 0.022, 0.038, 0.047, 0.002]. In the weaning success group, left ventricular ejection fraction (LVEF), velocity of mitralannulus in systolic (LatSa), maximum flow velocity of aortic valve (AV-Vmax), velocity-time integral (VTI), left ventricular global longitudinal strain (LVGLS), left ventricular global longitudinal strain rate (LVGLSr) were all increased on the day before ECMO weaning compared with the first day of ECMO support [LVEF: 0.40±0.05 vs. 0.28±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.62±1.02, AV-Vmax (cm/s): 104.81±33.98 vs. 64.44±16.85, VTI (cm): 14.56±3.11 vs. 7.96±1.98, LVGLS: (-8.95±2.59)% vs. (-5.26±1.28)%, LVGLSr (1/s): -0.48±0.11 vs. -0.29±0.09], whereas the ECMO flow was significantly reduced (L/min: 1.46±0.47 vs. 2.64±0.31), the differences were statistically significant (all P < 0.05). There was no significant difference in left ventricular functional parameters between the first day of ECMO support and the day before ECMO weaning in the weaning failure group. Compared with the weaning failure group, the weaning success group had higher LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr on the day before ECMO weaning [LVEF: 0.40±0.05 vs. 0.26±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.31±1.03, AV-Vmax (cm/s): 104.81±33.98 vs. 67.67±18.46, VTI (cm): 14.56±3.11 vs. 7.75±2.77, LVGLS: (-8.95±2.59)% vs. (-4.81±1.81)%, LVGLSr (1/s): -0.48±0.11 vs. -0.30±0.10, all P < 0.05] and lower ECMO flow (L/min: 1.46±0.47 vs. 2.20±0.62, P < 0.05). Conclusion:Bedside echocardiographic left ventricular function parameters (LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr) combined with clinical indicators (MAP, VIS, SCr, Lac, SpO 2) were helpful to evaluate the therapeutic effect of patients receiving VA-ECMO support and can provide important guiding value in the selection of VA-ECMO weaning timing and the judgment of prognosis.

Objective To evaluate the expression characteristics of calcium-binding protein S100A8 (Calgranulin A) and S100A9 (Calgranulin B) in lung squamous cell carcinoma and lung adenocarcinoma and their correlations with clinical data.Methods Twenty paired snap frozen cancer tissue and para-cancer tissue of lung cancer patients were collected form Jan 2013 to Dec 2014 in our hospital.And other 100 cancer paraffin wax specimen,20 para-cancer paraffin wax specimen were retrospectively collected from Jan 2007 to Dec 2009.The expressions of S100A8 and S100A9 were analyzed by PCR,Western blot and immunohistochemistry assay.The association between S100A8,S100A9 expression and patients clinical characteristics were analyzed.Results Positive rates of S100A8,S100A9 mRNA were 100.0％ for cancer tissue,significant higher than those of para-cancer tissue of 40.0％ and 85.0％ (P ＜0.05).Positive rates of S100A8,S100A9 protein were 100.0％ for cancer tissue,significant higher than those of para-cancer tissue of 20.0％ and 90.0％ (P ＜ 0.05).S100A8 positive expression rate was 28.6％ and 89.5％ for well differentiation and poor differentiation groups with statistical difference (P ＜ 0.05).The expression rate of S100A9 for stage Ⅰ positive was 73.0％ and the 100.0％ of S100A9 for stage Ⅲ/Ⅳ groups with statistical difference (P ＜ 0.05).Conclusion The expression levels of S100A8 and S100A9 in lung adenocarcinoma and lung squamous cell carcinona was higher than those in normal tissue,which were negatively correlated with the degree of differentiation of lung adenocarcinoma,and without correlation to the degree of differentiation of lung squamous cell carcinoma.The expression level of S100A9 was negatively correlated with tumor TNM stage.

ObjectiveTo investigate the expression and distribution of Toll-like receptor 4 (TLR4) in renal tissue of HBV associated nephropathy (HBV-GN) and its role in the pathogenesis and clinical manifestations of HBV-GN.MethodsRenal tissues were sampled from 48 HBV-GN patients confirmed by renal biopsy and 154 non-HBV-GN patients.The distribution of TLR4 in renal tissue and the relationship between the distribution of TLR4 and HBsAg were detected by immunohistochemistry.Integrating case record,correlations between the expression of TLR4 with clinical parameters including pathology,glomeruli,kidney tubules lesions,renal interstitial inflammatory infiltration and blood serum HBV were analyzed.ResultsTLR4 mainly distributed in the renal tubular epithelial cells and interstitial areas as brownish red and granular,which was in consistent with HBsAg distribution.The TLR4 positive rate and score in HBV-GN group were higher than those in non-HBV-GN group (P ＜ 0.05 ).TLR4 positive score was slightly higher in mesangial proliferative glomerulonephritis group and focal segmental glomerulosclerosis group,which had no significant difference (P ＞ 0.05).Kidney tubules lesions were strongly associated with TLR4 expression (r =0.748,P ＜ 0.001 ) which increased with aggravation of renal interstitial fibrosis ( r =0.569,P ＜0.001 ),tubular atrophy ( r =0.577,P ＜ 0.001 ) and inflammatory cell infiltration ( r =0.684,P ＜0.001 ).No obvious correlation with glomeruli lesions was observed ( r =0.293,P =0.053 ).Negative correlation could be seen between TLR4 and the renal function ( R2 =0.784),systolic blood pressure ( R2 =0.869),high sensitivity C-reactive protein (R2 =0.979) and urinary protein (R2 =0.615 ) by regression analysis.Other clinical parameters had no statistical significances.ConclusionsThe expression of TLR4 is abnormal in the renal tissue of HBV-GN patients,mainly in renal tubular epithelial cells and interstitial,which is consistent with the distribution of HBsAg.Its intensity is closely related with renal interstitial lesions,renal function changes and inflammatory cell infiltration.A speculation,that HBV can promote abnormal expression of TLR4 in renal tissues of HBV-GN which may be involved in the lesion progress of HBV-GN,is made upon our study.

Confronted with similar problems and difficulties in mainland,Taiwan reformed its medical education. The reform included setting up flexible program,formulating objective and detailed rotary guide,making the best of the resources,implementing medical humanities education based on the reality and forming unique cultivation system and mode. These measures can be taken as references for medical education reform in mainland.

Objective To explore the expression and role of Toll receptor 4(TLR4)in human proximal tubular epithelial cell line HK-2,infected by HBV. Methods The serum of HBV DNA copies between 107-108/ml was collected. Before and after infected by HBV DNA positive serum. the HK-2 cells' morphology and the expression of α-smooth muscle actin(α-SMA)were observed by microscopy and immunofluorescence, and the effects of different concentrations of lipopolysaccharides(U)S.TLR4-stimulating factor)and CLI-095(TLR4 Inhibitor)on the proliferation rate of HK-2 cells were observed by MTT assays. After HBV serum and 10μg/ml LPS and 5μl/ml CLI-095 acted on HK-2 cells,TLR4 protein expression was measured by immunofluorescence and Western-blotting assay, and HBsAg and HBeAg in cell culture medium were detected by ELISA. and HBV DNA copies by fluorescence quantitative PCR. Results The longer HBV infected HK-2 cells, the more irregular of the cells' shape, the fewer number of the cells were left. But compared with HBV infected after 24 hours, α-SMA was more expressed after HBV infected 12 hours. After infected by HBV serum in 24 hours.HK-2 cells' proliferation rate was positively correlation in a dose range of LPS, but was negatively correlated with the CLI-095(P＜0.05＝.The levels of HBsAg and HBeAg in cell culture medium were largest when the LPS concentration was at 10μg/ml and CLI-095 at 5μg/ml.The expression of TLR4 significantly increased in HK-2 cells treated with LPS compared with those with CLI-095.but HBV DNA levels and HBsAg and HBeAg expression levels were lower. Conclusions HBV infection may promote cell transdifferentiation and cell injury. The stimulation of HK-2 infected with HBV by LPS may upregulate the expression of TLR4 and reduce the copies of HBV DNA.