Objective To explore the effect of the C gene truncated HBV mutant on HBV replication. Methods The C gene truncated HBV vector pHBV-?C was constructed through the molecular clone in vitro,and then transfected transiently into HepG2 cell.The expression of S protein was assayed by ELISA and SDS-PAGE Western blot.After co-transfection with pHBV-?C and wild HBV genome adwR9 into HepG2 cell the DNA was detected quantitatively by real-tine fluorescence quantitative PCR in the culture medium and the cell. Results There was no significant difference in expression of S protein assay by ELISA and Western blot.The DNA of the cotransfected group with pHBV-?C and adwR9 was lower than that of control group in the culture medium and the cell. Conclusions(C gene) truncated HBV mutant can cause the reduction of HBV replication.

Objective:To investigate the significance of T helper type 17 cells(Th17)/T regulatory cells (Treg) in the prognosis of chronic hepatitis B(CHB) by comparing the changes of Th17 cells ,Treg cells and their rate under different therapy in chronic hepatitis B patients.Methods: 40 patients infected with chronic hepatitis B(CHB) were divided into chronic hepatitis B without antiviral(CHBWA) group and chronic hepatitis B antiviral(CHBA) group depending on whether nucleoside antiviral was applied,with 20 cases each,10 cases in asymptomatic hepatitis b carriers(AsC) group.10 cases in health control (HC) group.Flow cytometer(FCM) was adopted to detect peripheral blood Th17 and Treg cell frequency in CHB patients at 0,1 month in CHBWA group ,at 0,1,3 month in CHBA group,AsC patients and HC cases ,then calculating Th17/Treg ratio and observing the dynamic changes at different disease states.Results: After the liver-protecting treatment in CHBWA patients,the Th17/Treg ratio at 1 month(0.39±0.11)decreased compared to 0 month(1.20±0.26)rapidly(P<0.01),and the ratio nearly reached the level of AsC and HC group (P>0.05).Ratio of Th17/Treg and ALT had good positive correlation (r=0.709,P=0).In CHBA patients group,after the liver-protecting and antiviral treatment,patients′ Th17/Treg ratio decreased rapidly in the first month(0.73±0.32) and third month(0.76±0.44),and there′s statistical difference compared with 0 month(1.18±0.27,P<0.01),but there was no significant change of Th17/Treg ratio after being treated for 3 months compared with 1 month,and the Th17/Treg ratios at 0,1,3 month were higher than that in AsC group and HC group(P<0.01).Ratio of Th17/Treg and ALT have good positive correlation (r=0.500,P=0.000),also the ratio of Th17/Treg and HBV DNA were positively correlated (r=0.345,P=0.007).Comparison of peripheral blood Th17,Treg cells frequency,Th17/Treg ratio between CHBWA and CHBA group at 0 month had no statistical significance(P>0.05),while the Th17/Treg ratio in CHBA group was higher than CHBWA group(t=4.471,P<0.01) after 1 month of treatment.Conclusion: In the CHB patients,antiviral treatment will influence the Th17,Tregs cells frequency,resulting the change of Th17/Treg ratio,but also the ratio may be associated with virus removal.

Objective:To investigate the protective effect of intratracheal transplantation of different dose of human umbilical cord mesenchymal stem cells (MSCs) in rats with acute lung injury induced by severe burns.Methods:Seventy-five male Wistar rats were randomly divided into five groups:Sham(group A),Saline group(group B) and different doses of hUMSCs transplantation groups(C,D and E).The dosage ofhUMSCs was 1 × 105,5 × 105 and 1 × 106 respectively.Rats inflicted by 50 ％TBSA Ⅲ degree scalding employed as the model.After modeling,rats in group B and transplantation groups were immediately fluid resuscitated.Transplantation groups were intratracheally administered different dose hUCMSCs (0.2 mL),and group B were given normal saline in the same dose intratracheally.The lung tissue samples were collected on day 1,day 3 and day 7 after administration.HE staining was used to observe the pathological changes of lung tissue.MPO and CD68 immunohistochemical staining were used to observe the positive expression of neutrophils and macrophages in lung tissue.Results:Lung pathology showed that alveolar cavity was clear,alveolar structure integrity,occasionally a small amount of inflammatory cells of group A at each time point.At 1 day after scald,group B and the transplantation group (group C,D,E)the alveolar septum was thickened,and there was visible pulmonary capillary hyperemia,as well as a large amount of inflammatory cell infiltrations in the pulmonary capillaries and alveolar space.At 3 day,group B and the transplantation group alveolar structural damage,pulmonary hemorrhage and inflammatory cell infiltrations were better than those in 1 day.Compared with group B,the alveolar structure was clear and the septum was thinner,but there was no significant difference between the transplantation groups.On the 7 day after scald,the lung injury in the transplanted group was significantly less than group B,and the recovery of the injured lung tissue in E group was the most obvious.The number of the MPO positive cells increased significantly on the first day after scald (P ＜0.05) compared with group A,but there was no significant difference between the two groups.Compared with B group,the number of positive cells in transplantation group was significantly reduced at 3 and 7 day after scald,(P＜0.05),and the number of positive cells in group E was significantly lower than other groups (P＜0.05).CD68 staining showed a significant increase in positive cells in each group on day 1 (P＞ 0.05).The number of positive cells decreased in 3 day after transplantation (P＜0.05),but there was no significant difference between the transplantation groups.The number of positive cells in transplantation group was significantly lower than group B (P＜0.05) after 7 day.Compared with group C and D,there was significant difference in group E (P＜0.05).Conclusions:Intratracheal transplantation of different dose hUCMSCs have protective on severe burns induced acute lung injury models;the protection mechanisms may be that the hUCMSCs transplantation can inhibit the invasion of the inflammatory cells in lung tissues,and the optimal dosage is 1 × 106.

Objective To investigate the anticancer effects and detailed mechanisms of Saikosaponin D (SSD) in human hepatoma HepG2 cells. Methods Cell proliferation and apoptosis were tested by MTT assay and Annexin-V/PI assay respectively. The expressions of CCAAT enhancer binding protein β(C/EBPβ) and p53 were detected by RT-PCR and Western blotting. Results SSD inhibited cell proliferation in a dose-dependent manner and induced apoptosis at the concentration of 5.0mg/L. SSD significantly increased the mRNA and protein levels of C/EBPβ and p53 in a dose-dependent manner. Conclusion SSD exerts its anticancer effect by inhibiting cell proliferation and inducing apoptosis partly through C/EBPβ-p53 signal pathway in HepG2 cells.

Correct implementation of laparoscopic total gastrectomy with D2 lymph node dissection depends on the understanding of anatomical features of peripancreatic space, the landmark of pancreas and blood vessels, and the diverse perigastric vascular anatomy and standardized surgical procedures designed according to the regional distribution of lymph nodes.From September 2006 to November 2009, laparoscopic total gastrectomy with D2 lymph node dissection surgery were completed in 12 cases in the Nanfang Hospital. A reasonable anatomy method and a simple, effective surgical procedure of laparoscopic D2 lymph node dissection were introduced.

Laparoscopic pancreas- and spleen-preserving splenic hilar lymph nodes dissection is still difficult to accomplish,which restrains its application in total gastrectomy for advanced proximal gastric cancer.Based on our anatomical understanding of pre- and retropancreatic spaces,features of splenic vessels and distribution of perigastric lymph nodes,we combined the characteristics of laparoscopic surgery and developed a novel technique for laparoscopic pancreas- and spleen-preserving splenic hilar lymph nodes dissection through retropancreatic space.The key lies in mobilization of the splenic pedicle through retropancreatic space,dissection of peri-vascular lymph nodes in sequence of trunk-to-branch,in-sheath vascularization of the splenic vessels.From August 2009 to December 2010,this technique was performed on 6 patients in Nanfang hospital.The initial results suggested that this technique could be safe and feasible for skillful surgeons.Further studies on the application of this technique in total gastrectomy for advanced proximal gastric cancer would be needed.

Objective To evaluate the effect of dexmedetomidine administered locally on the median effective concentration ( EC50 ) of ropivacaine for paravertebral nerve block ( PVNB) . Methods Forty?eight ASA physical status Ⅰ or Ⅱ female patients, aged 20-64 yr, with body mass index<24 kg∕m2 , scheduled for elective unilateral segmental mastectomy under PVNB, were randomly divided into 2 groups ( n=24 each) using a random number table: ropivacaine group ( group R) and ropivacaine mixed with dexmedetomidine group ( group RD) . PVNB was performed at T4 on the operated side guided by ultrasound and nerve stimulator. Ropivacaine 20 ml and a mixture of ropivacaine and 20 μg dexmedetomidine 20 ml were injected locally in group R and group RD, respectively. The concentration of ropivacaine was determined by up?and?down sequential allocation. The initial ropivacaine concentration was set at 0. 35%, and the ratio between the two successive concentrations was 1. 2. The EC50 and 95%confidence interval of ropivacaine were calculated using Dixon?Massey method. Results The EC50 ( 95%confidence interval) of ropivacaine was 0.27% (0.23%-0.30%) and 0.22% (0.18%-0.25%) in group R and group RD, respectively. Compared with group R, the EC50 of ropivacaine was significantly decreased by 19% in group RD. Conclusion Small dose of dexmedetomidine administered locally can significantly enhance the efficacy of PVNB with ropivacaine.

Objective To study the alternations of regulatory T cells in early HIV infected patients and its association with disease progression.Methods Fifty-one untreated HIV infected patients were enrolled and divided into 3 groups according to their infection time and CD+4 T cell levels(30 early HIV infected patients,15 typical progressors,6 AIDS patients).Twenty normal controls were enrolled.There were no significant differences between the age and sex among four groups.Blood was drawn by venipuncture from each subject in EDTA tubes and the levels of CD+4 CD+25 Foxp3 + regulatory T cells were detected by FACSAria flow-cytometry.Spearman correlation was used to detect association between CD+4 CD+25 Foxp3 + regulatory T cells and the absolute CD+4 T cells,viral load and activation of T cells.Results The levels of CD+4 CD+25Foxp3+ regulatory T cells showed the tendency of increasing tendency from normal control to early HIV infected patients,asymptomatic HIV infected patients and AIDS patients.Early HIV infected patients was significantly lower than that in AIDS group [3.79(2.11 - 5.43) % vs 8.09(4.90 - 8.90) %,Z = - 2.29,P = 0.022].The levels of CD+4 CD+25 Foxp3 + Treg cells were associated with viral set point(r = 0.479,P =0.038) and inversely associated with CD+4 T cells(r = -0.455,P =0.011) and closely associated with HLA-DR expression on CD+3 T cells(r = 0.533,P = 0.002).Conclusions The ratio of CD+4 CD+25 Foxp3 +regulatory T cells of early HIV infected patients was significantly increased and associated with viral set point and CD+4 T cell counts,which indicate that alternation of regulatory T cell may be an important factor contributing to the disease progression in early HIV infection.

Objective: To explore the impact of ticagrelor on platelet aggregation in patients with acute coronary syndrome (ACS) after percutaneous coronary intervention(PCI). Methods: A total of 98 ACS patients received PCI in our hospital from 2015-01 to 2015-12 were enrolled. The patients were randomly divided into 2 groups: Clopidogrel group, the patients received oral clopidogrel 300mg at first time and then maintained by 75mg/qd, n=48 and Ticagrelor group, the patients received oral ticagrelor 180mg at first time and then maintained by 90mg/bid, n=50. All patients were treated for 12 months.The level of vasodilator stimulated phosphoprotein (VASP) phosphorylation and platelet reactivity index (PRI) at pre-medication and 24h, 7 days and 1 month after PCI were detected; major adverse cardiovascular events (MACE) and bleeding events were recorded within 1 month after PCI, the incidence of platelet aggregation, MACE and bleeding events were compared between 2 groups.Results: The baseline information and PCI condition were similar between 2 groups, P>0.05. The overall average PRI was different between 2 groups, P<0.001 and PRI at each time point was different between 2 groups, P<0.001, different group and time point had interactive effect on PRI, P<0.001. Compared with Clopidogrel group, Ticagrelor group had the lower ratio of PRI≥50% at different time points after PCI, P<0.001. The incidence of MACE and bleeding event were similar between 2 groups within 1 month after PCI, P>0.05. Conclusion: Ticagrelor was superior toclopidogrel for anti-platelet aggregation in ACS patients after PCI, it didn't increase bleeding events.

Objectives To assess the plasma levels of acyl ghrelin (AG) and dys-acyl ghrelin (DG) in chronic kidney disease (CKD) and hemodialysis (HD) patients and analyze their relationships with different stages of CKD and hemodialysis. Methods Forty-six CKD stage 1-5 patients and 15 hemodialysis patients were enrolled into the study. Body weight, height, hemoglobin, biochemical parameters, inflammatory parameters, preprandial, postprandial and 3 hours after hemodialysis plasma AG and DG levels were measured. Appetite and food intake were assessed. Body mass index (BMI), and estimated glomerular filtration rate (eGFR) were calculated. Results There were no significant differences in BMI, SGA, appetite, food intake and malnutrition among CKD patients of different stages. eGFR was declining with the progression of CKD stages and patients received a three-week hemodi-alysis. Compared with that in CKD stage 1-2 patients, the level of preprandial and postprandial DG was remarkably increased in stage 3-5 patients (P<0.01). The level of DG was significantly decreased after a standard breakfast in CKD patients (P<0.01). CKD stage was positively correlated with preprandial (r=0.31, P<0.05)a nd postprandi-al DG (r=0.34, P < 0.05), TNF-α (r=0.33, P < 0.05), IL-6 (r=0.40, P < 0.05), leptin (r=0.34, P < 0.05), and age (r=0.41, P<0.05). CKD stage was also highly and positively correlated with the proportion of preprandial and postprandial DG (r=0.61, P<0.01;r=0.55, P<0.01). Multivariate partial-correlation analysis showed that CKD was independently associated with the proportion of preprandial and postprandial DG (r=0.55, P < 0.01; r=0.43, P < 0.01).There was no decrease in AG postprandially, nor any changes in AG resultant from dialysis (P > 0.05);levels of DG decreased slightly postprandially and were markedly decreased by hemodialysis (P<0.01), even lower than those seen postprandially in CKD stage 1-2;Both preprandial and postprandial DG were negatively correlated with serum albumin levels (r=-0.64, P < 0.05; r=-0.59, P < 0.05), while there was no correlation between AG and serum albumin levels. Conclusions There is a strong and independent correlation of DG with CKD stage. Postprandial suppression of ghrelin is impaired with reduced renal function. Hemodialysis removes DG but not AG.