Objective To observe the transport of hepatitis B virus (HBV)-infected peripheral blood mononuclear cells (PBMC)through placental barrier set up by choriocarcinoma trophoblast cells (Bewo cells),and to explore the biological role of PBMC as a carrier for HBV transport.Methods Bewo cells and PBMC were cultured and their proliferation and activity were detected by cell counting kit (CCK)-8.One hundred μL serum containing 5 ×10 6 copy/mL HBV DNA was used to infect PBMC,and cells infected with HBV were labeled by fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE).A co-culture model of Bewo cells and HBV-infected PBMC was set up by transwell chamber. The migration of HBV-infected PBMC was detected by flow cytometry.Realtime fluorescence quantitative polymerase chain reaction method was used to detect HBV DNA contents of PBMC under transwell chamber.Results PBMC and Bewo cells proliferated at around 24 h and entered into growth stagnation at around 120 h.The contents of PBMC labeled by green fluorescent at 0,12,24 and 48 h during co-culture under chamber were (0.445 ±0.021)%,(21 .180 ± 4.653 )%,(34.830 ± 7.156 )% and (64.185 ± 3.161)%,respectively.The amount of PBMC marked green fluorescence increased over prolonged incubation time (F =68.983,P =0.001 ).PBMC HBV DNA contents at 24 and 48 h of co-culture under chamber were (1.925±0.431)×103 copy/mL and (2.565 ±0.361)×103 copy/mL,respectively,indicating that PBMC under chamber were infected with HBV.Conclusions PBMC may be a target for HBV infection in extrahepatic tissues.Placental trophoblastic barrier built by transwell chambers may provide new ideas to investigate HBV transmission across the placenta in vitro .

Objective To investigate the infiltration of dendritic cell (DC) in Vocal cord polyp, laryngeal leukopla-kia and glottic squamous cell carcinoma,and to observe laryngeal mucosal lesions of the state of the immune microenviron-ment, and to research significance of DC on the development of glottic squamous cell carcinoma. Methods The infiltration of S-100+and CD83+DC in 20 cases of Vocal cord polyp, 47 cases of laryngeal leukoplakia, 45 cases of glottic squamous cell carcinoma tumor and 20 cases of laryngeal normal mucosa were examined using immunohistochemistry. Results The num-ber of S-100+and CD83+DC were significantly higher in glottic squamous cell carcinoma, laryngeal leukoplakia and vocal cord polyp than that in laryngeal normal mucosa (P<0.05). The number of S-100+and CD83+DC were higher in laryngeal leukoplakia than that in glottic squamous cell carcinoma (P<0.05). The number of S-100+and CD83+DC in mild dysplasia and moderate dysplasia were less than that in severe atypical hyperplasia (P<0.01). In glottic squamous cell carcinoma, S-100+ and CD83+DC with poor-differentiated was significantly less than that with well-differentiated (P < 0.01). Conclu-sion Changes in the number of dendritic cell was found in vocal cord polyp, laryngeal leukoplakia and glottic squamous cell carcinoma, which indicated that there were an abnormal immune status. Changing of dendritic cell in laryngeal mucosa plays an important role in laryngeal cancer development.

Objective To explore the relationship of hepatitis B virus surface antigen(HBsAg)infec-tion and family history of hepatocellular carcinoma(HCC)with age at primary liver cancer. Methods Totally 1 359 cases of primary liver cancer were enrolled. Their data of sex,HBsAg status and family history informa-tions of liver cancer were analyzed on the associations with diagnosis age. Results Of the 1 359 cases,1 053 were males and 306 were females,their average age at diagnosis was(54. 02 ± 10. 47)years(20-84 years). For HBsAg positive cases,the average age at diagnosis was 51. 99,significantly younger than that of HBsAg negative cases(61. 23),t = 13. 51,P = 0. 000. Cases with family history of HCC were diagnosed at a signifi-cantly earlier age than those without family history(52. 53 vs 55. 23,t = 4. 389,P = 0. 000). In HBsAg posi-tive cases,the average age at diagnosis showed a significant difference not only between males and females (51. 18 vs 54. 89,t = 5. 353,P = 0. 000),but also between cases with family history and cases without family history(51. 33 vs 52. 62,t = 2. 233,P = 0. 026). In HBsAg negative cases,the average age at diagnosis of males and females were 60. 83 and 62. 45 respectively(t = 1. 126,P = 0. 261). The average age at diagnosis of cases with family history and cases without family history were 59. 58 and 61. 92 respectively(t = 1. 728,P =0. 085),both showed no significant difference. Conclusion Cases of primary liver cancer with positive-HBsAg are diagnosed averagely 9. 24 years younger than those with negative-HBsAg in Qidong. Sex and family history of HCC significantly advance hepatocarcinogenesis only in HBsAg positive individuals,not in HBsAg negative individuals.

Objective To observe the clinical efficacy of bosentan in treatment of severe pulmonary hypertension related to congenital heart disease (CHD-PAH) .Methods 5 patients with severe CHD-PAH patients received bosentan therapy, then pulmonary artery pressure, pulmonary vascular resistance (PVR), 6min walk test,right ventricular end-systolic diameter (RVSD) changes were observed and statistically analyzed after six months medication. Results Pulmonary arterial systolic pressure (sPAP) was significanfly decreased from (96±11) mmHg to (86±10) mmHg, <0.01.pulmonary arterial diastolic pressure (dPAP) was significanfly decreased from (56±10) mmHg to (46±9) mmHg ( <0.01),pulmonary arterial mean pressure (mPAP) was significanfly decreased from (73 ±11) mmHg to (59 ±10) mmHg ( <0.05), pulmonary vascular resistance was significanfly decreased from (17.8±1.9) Wood to (13.1±1.7) Wood (<0.01) . 6min walk test was improved from (136±40) m to (198±55) m, <0.01.right ventricular end-systolic diameter significanfly decreased from (40±5) mm to (36±6) mm after 6 months therapy ( <0.05) . Conclusion Bosentan can decrease pulmonary arterial systolic pressure, improve exercise tolerance, improve right ventricular function in patients with severe CHD-PAH.

Objective Sirolimus ( SRL) is a potent immunosuppressive drug used to prevent acute allograft rejection after re-nal transplantation.Nevertheless, the occurrence of proteinuria has recently been recognized among patients treated with SRL-based therapy.The aim of this study was to investigate the therapeutic effect of tripterygium wilfordii hook F ( T II) on proteinuria caused by SRL in renal transplant recipients who were treated by trilogy immunosuppressive therapy of sirolimus combined with mycophenolate and hormone. Methods 52 recipients were divided into 2 groups randomly:TⅡgroup (n=27) and valsartan group (n=25).The TⅡgroup was administered 1 mg/kg/d, and the valsartan group 80-160 mg/d for consecutive 12 months.Based on primary trilogy immu-nosuppressive therapy of sirolimus combined with mycophenolate and hormone, the dosage of sirolimus was adjusted according to the target concentration 6-10 ng/ml( ELASA approach) and mycophenolate was administered 750 mg twice per day, adjusting dosage ac-cording to the mycophenolate AUC 0-12 level(35-45 mg· h/L).The evaluation of therapeutic effect includes: complete remission, proteinuria decreased by>50%; partial remission, proteinuria decreased by 20% to 50%; ineffective, proteinuria decreased by<20%. Results During the 12 month follow-up, the total effective rates in the TⅡgroup and the valsartan group were 95.2%and 86.7%respectively, in which the TⅡ group decreased more significantly (P<0.01).The total cholesterol level and triglyceride level in TⅡgroup were obviously lower than those in valsartan group(P<0.01). The total cholesterol level and triglyceride level in valsartan group increased ([6.60±0.2]mmol/L vs [7.11±1.13]mmol/L, [2.47± 1.48]mmol/L vs [2.49±0.32] mmol/L).The serum protein level in TⅡ group was obviously higher than that in valsartan group ([41.1±1.2]g/L vs [37.9±4.2]g/L, P<0.05).At 3 month, 6 month and 12 month follow-up, the average serum creatinine levels in TⅡgroup were obviously lower than those in valsartan group ([1.5±0.4]mg/dl vs [1.6±0.3]mg/dl, P<0.05), ([1.3±0.3]mg/dl vs [1.8±0.5]mg/dl, P<0.05), ([1.1±0.4]mg/dl vs [2.1±0.5]mg/dl, P<0.05).The incidence rate of adverse reaction in valsartan group was obviously greater compared with TⅡgroup( P<0.05) . Conclusion Both tripterygium wilfordii multiglycosides and valsar-tan can reduce proteinuria caused by SRL in renal transplant patients,while the application of TⅡhas more remarkable effect.

Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.

Objective Little research has been done on the risk factor analysis of BK virus(BKV) infection in renal transplant recipients in Chinese population.The article aimed to investigate BKV infection and analyze its risk factors in renal transplant recipients in China.Methods Renal transplant recipients who had received the detection of BKV DNA in urine and blood samples in Nanjing General Hospital from June 2015 to July 2016 were selected, while the patients with uremia hemodialysis and healthy living donors were included as control group.According to the detection results of BKV DNA in urine and blood samples, renal transplant recipients were divided into BKV DNA positive group(n=89, positive urine or blood and urine BKV DNA) and BKV DNA negative group(n=359, negative blood and urine BKV DNA).Analysis was made on BKV infection in renal transplant recipients in order to investigate the effects of factors including clinical condition, postoperative complications and immunosuppressive regimen on BKV infection.Results The positive rate of BKV DNA in urine samples of renal transplant recipients was 19.9%, which was higher than those of patients with dialysis and healthy living donors(6.3% and 4.2% respectively, P<0.001).Multivariate logistic regression analysis showed BKV infection was associated with pulmonary infection(OR[95%CI], 3.468[1.227-9.802];P=0.019) , acute rejection (OR[95%CI], 2.645[1.142-6.127];P=0.023), and FK506 (OR[95%CI], 2.408[1.104-5.254];P=0.027).Conclusion The incidence of BKV infection in renal transplant recipients increases significantly.Pulmonary infection, acute rejection and FK506-based immunosuppressive regimen are risk factors leading to BKV infection.

Objective BK virus-associated nephropathy ( BKVAN) after kidney transplantation is a key factor that influence the prognosis of transplant kidney .To our knowledge , it is believed to be associated with immune suppression .We observed the cura-tive effect and influencing factorsof anti-rejection scheme that Leflunomide was administered instead of Mycophenolate Mofetil ( MMF) on transplant kidney BKVAN .. Methods This study included 15 kidney transplant recipients with pathologically confirmed BKVAN in Nanjing General Hospital of Nanjing Military Region form March 2007 to March 2013 .Leflunomide was administered instead of Myco-phenolate Mofetil ( MMF) .Serum creatinine level , renal allograft loss rate and side effects of leflunomide were monitored after medica-tion switch.The patients were divided into two groups , which were renal allograft loss group and renal allograft survival group , for fur-ther analyses . The differences between each groups in clinical characteristics as well as histochemical features of the transplanted kidneys were analyzed to determine the cause of renal allograft loss in patients with BKVAN . Results Six patients experienced renal al-lograft loss after switching to leflunomide and needed hemodialysis , and 9 patients had stable renal allograft function , renal allograft loss rate was 40.0%.Hyperuricemia occurred in 8 patients in the period before the medication switch and in 5 patients after the switch;a decrease in blood white cell orplateletcount was found in 2 patients during both periods;an increase in Alanine aminotransferase ( ALT) level occurred in one patient after the medication switch .There were no statistically significant differences in any of the above parame-ters before and after the medication switch.Compared to allograft survival group, serum creatinine level[(1.80 ±0.53)mg/dL vs (2.74 ±0.58)mg/dL, P=0.007], the number of B lymphocytes [(206.44 ±144.96) vs (439.67 ±267.77), P=0.047] and CD68[(588.44 ±271.80) vs (944.67 ±259.32), P=0.025] in renal allograft tissue were significantly higherin the allograft loss group. ConclusionLeflunomide is a safe and effective medication for BKVAN .Patients with significantly increased serum creatinine level might have a poorer prognosis .Significantly increased B lymphocytes and CD 68 cells in renal allograft tissue might indicate a poor prognosis.

Objective To analyze the management of clinical pathways ( CP) in China. Methods Cross-sectional questionnaire surveys of 51 public hospitals with CPs in place in Shanghai, Hubei province and Gansu province were conducted from March to May of 2015. Results Among the 51 public hospitals with CPs, 48 ( 94. 1%) of them organized training on CPs, 48 ( 94. 1%) of them monitored CPs′implementation, and 40 (78. 4%) applied incentives for CPs′ implementation. But there were some issues and difficulties encountered in CPs′ implementation. Conclusions Comprehensive measures are necessary to improve the management of CPs at public hospitals of China.

Objective To analyze the implementation of clinical pathways ( CP) at public hospitals at different levels and in different regions in China. Methods The status of CPs′ implementation at 54 public hospitals in Shanghai, Hubei province and Gansu province was surveyed by questionnaires from March to May of 2015. Results 51 (94. 4%) of the surveyed public hospitals put in place clinical pathway(s), where the average CPs implemented were 45 and the average percentage of the cases using CPs was 52. 7%. There were great variations among these hospitals. In addition, the common diseases with definite diagnostic and treatment options were found with the highest implementation rates of CPs at such hospitals. Conclusions CPs are implemented widely at public hospitals of China, yet enhanced implementation strategies are expected to further CPs′adoption.