Objective To compare the clinical efficacy and safety of endoscopic papillary balloon dilation (EPBD) and endoscopic papillary balloon dilation (EPBD) combined with small endoscopic sphincterotomy (SEST) in treatment of large choledocholithiasis. Methods 78 patients with large choledocholithiasis from January 2014 to December 2015 were randomly divided into EPBD group, and combination treatment group. The level of bilirubin, transaminase, alkaline phosphatase (ALP) before and after the operation, the success rates of stone removal, serum amylase of 24 h after the operation, the operation times of endoscopic retrograde cholangiopancreatography (ERCP) and whether complicated with postoperative pancreatitis were compared between the two groups. Results There was no significant difference of the success rates of stone removal, serum amylase of 24 h after the operation, the operation times of ERCP between the two groups. The level of bilirubin, transaminase, alkaline phosphatase (ALP) was declined after EPBD or EPBD and SEST, and the results of the two groups had no statistical significance (P > 0.05). Also there was no significant difference of the incidence of postoperative pancreatitis, postoperative bleeding and postoperative hyperamylasemia between the two groups (P > 0.05). Conclusion EPBD is worthy of promoting because it has a similar clinical efficacy and safety to EPBD and SEST in treatment of large choledocholithiasis.

Objective: To investigate the role of the recombinant S protein of SARS-CoV in the induction of chemokine IP-10 and other cytokines in airway epithelial cells and immunocytes. Methods: Using insect-baculovirus expression system and Nickel affinity Magnet Beads, S protein of SARS-CoV was produced and then used to stimulate cultured human bronchial epithelial cells (16HBE), human peripheral blood mononuclear cells(PBMC), human peripheral blood monocytes and alveolar macrophages. The levels of IP-10 and the cytokines involved in immunoreaction in response to virus infection were detected in the supernatants of those cells cultured with the S protein by liquid chip system. Results: Under normal condition, no detectable IP-10 was found in 16HBE. A high level of IP-10(79.97? 13.81) pg/ml was detected in the 16HBE 12 hrs after being treated with the S protein, and the induction of IP-10 by S protein displayed at a significant quantity-effect reaction, but not in PBMC, monocytes and alveolar macrophages. In contrast, IFN-? was able to induce the production of IP-10 in either 16HBE or the immunocytes. Conclusion: 1.S protein of SARS-CoV can induce a high level of IP-10 in lung epithelial cells at early stage after the virus infection, which may initiate the process of the immune damage in the lung. 2. S protein of SARS-CoV induces the production of IP-10 by a way of IFN-? independent.

OBJECTIVE To investigate the relation of expression and significance of IL-25,IL-12 and IgE in children with allergic rhinitis.METHODS The concentration of IL-25,IL-12 were measured in 34 children with allergic rhinitis and 34 health control subjects by enzyme-linked immunosorbent assay(ELISA),and compare to them and relationship.RESULTS The concentrations of IL-25,IL-12,IgE in periodicitical allergic rhinitis(AR)were(42.64? 11.77)ng/L,(38.52?8.32)ng/L,(563.2?141.7) kU/L,respectively.And in persistence AR they are(57.51?5.36)ng/L,IL-12(29.31?11.69)ng/L,(625.6?104.3)kU/L.However,in control group,the concentrations of IL-25,IL-12 and IgE were(33.42 ?9.78)ng/L,(49.31?11.69)ng/L(209.7?113.2) kU/L,respectively.There were significant difference in periodicitical AR and persistence AR,periodicitical AR and control group,persistence AR and control group(P

Objective To investigate the effect and safety of Tongqiaoxingnao soup applied in elder severe craniocerebral injury stasis resistance qing qiao patients.Methods 40 cases with severe craniocerebral injury stasis resistance qing qiao were enrolled in the study,and they were divided into two groups randomly,each group had 20 cases.Patients in the monotherapy group received routine western medicine treatment,while the combination group was added the treatment of Tongqiaoxingnao soup,200mL per day for 4 weeks.GCS scores and the persistent time of coma were analyzed.After treatment for 4,8,14 days of intracranial pressure and cerebral edema were recorded and analyzed,the total effective rate and the incidence of adverse reactions were compared between two groups.Results GCS score of the combination group was (13.7 ±4.2)points,and it was obviously higher than the monotherapy (9.2 ±3.5)points (t =2.86,P <0.05).The coma duration of the combination group was (6.5 ±1.4)d,and it was obviously shorter than the monotherapy group (10.2 ±2.5 )d (t =2.86,P <0.05 ).The amount of intracranial pressure increased of combination group were 10 cases and 8 cases,which were significantly less than the monotherapy after 8 and 14 days with 16 cases and 15 cases (χ2 =3.96,5.01,all P <0.05).Total effective rate of combination treatment group was 75.0%,which was significantly higher than 40.0% of monotherapy group (χ2 =5.01,P <0.05).The occurrence of adverse reactions such as bleeding,electrolyte imbalance,hypoxemia,and the acid -base imbalance had no significant differences between the two groups (P >0.05 ).Conclusion Tongqiaoxingnao soup applied in elder severe craniocerebral injury stasis resistance qing qiao patients has clinical curative effect,can shorten the coma time,reduce intracranial pressure and relieve cerebral edema,and has high security,it is worthy of populari-zation and application.

The B.subtilis ywtD gene,encoding a ?-polyglutamic acid(?-PGA)depolymerase,was amplified from the genome of B.subtilis NX-2 by PCR.The comparability between the cloned ywtD gene sequence to the reported sequence is high to 99.0%.Only one of the substituted nucleotide base caused the change to the amino acid sequence.The recombinant plasmid pET-15b-ywtD was then transformed into E.coli Rosetta(DE3)and the ywtD gene product could be expressed with the induction of 0.5mmol/L IPTG.The YwtD protein exhibited a remarkable activity in ?-polyglutamic acid degradation.The molecular weight of ?-PGA could be reduced from 700kDa to 20kDa after 72h through the enzymatic hydrolysis and consequently trended to be constant.

Objective To establish a cellular model for the expression of the C-type lectin dendritic cellspecific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN),and to provide a basis for the functional analysis of DC-SIGN.Methods The cDNA of DC-SIGN was obtained via PCR,and cloned into the eukaryotic expression vector porcine cytomegalovirus-enhanced green fluorescent protein (PCMV-EGFP) with EGFP at the N terminal of DC-SIGN.Then,the recombinant PCMV-EGFP-DC-SIGN plasmid was transfected into HEK293T cells followed by the detection of DC-SIGN expression using PCR,Western blot and flow cytometry.Confocal microscopy was performed to localize the expression of DC-SIGN-EGFP and visualize the recognization and internalization of the Derp2 allergen by DC-SIGN.Results The recombinant fluorescent fusion protein-expressing plasmid was successfully constructed.Both PCR and Western blot confirmed the expression of DC-SIGN.Flow cytometry showed that the expression of DC-SIGN was increased by approximately 50％ in HEK293T cells transfected with the recombinant expression plasmid compared with those untransfected.As confocal microscopy showed,the green fluorescence-labelled DC-SIGN was located on the cell membrane,which could bind to the red fluorescence-labelled antigen Derp2 and internalize it into the cells.Conclusions The recombinant DC-SIGN-EGFP fusion protein is characteristically located on the surface of 293T cells,which can be recognized by the DC-SIGN-specific antibody and is capable of internalizing the allergen Derp2,and may serve as an ideal cell model for further studies on molecular function of DC-SIGN.

Objective To synthesize herpes simplex virus (HSV) envelope glycoprotein gC using gene engineering techniques,and to verify its expression.Methods Two separate parts of the HSV envelope glycoprotein gC,i.e.,GC-F and GC-R,were respectively synthesized.The GC-F and GC-R genes were synthesized,subcloned into the expression vectors pSumo-Mut (containing recognition sequences for endonucleases Stu1 and XhoI) and pCzn1 (containing recognition sequences for endonucleases NdeI and XhoI) respectively to form the recombinant plasmids pSumo-Mut-GC-F and pCzn1-GC-R.E.coli BL21 Arctic Express (DE3) cells were transformed with the two recombinant plasmids separately.Isopropyl thiogalactoside (IPTG) was used to induce the expression of target protein which was subsequently purified by nickel affinity chromatography.Finally,Western blot was performed to verify the reactivity of the synthesized protein with the sera of HSV-1-positive patients.Results Both GC-F and GC-R genes were synthesized by a total gene synthesis method.As sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) showed,the fusion proteins were mainly distributed in the sediment layer.The purity of GC-F and GC-R proteins was over 80％ after purification by affinity chromatography.Western blot showed that both of the proteins were reactive with anti-HSV-1 antibody-positive sera.Conclusions Fusion expression vectors have been constructed for the gC protein,and IPTG successfully induces its expression.Moreover,the resulting proteins could react with anti-HSV-1 antibody-positive sera,and may serve as an ideal experimental material for next functional study.

Objective To investigate the effect of polyphenol of salvia miltiorrhiza polyphenols combined with collagen sponge in the treatment of diabetic foot.Methods A total of 115 patients with type 2 diabetic foot were randomly divided into treatment group (55 cases) and control group (60 cases).Salvia miltiorrhiza polyphenols and collagen sponge treatment group were treated with conventional treatment plus type 2 diabetes treatment of salvia miltiorrhiza polyphenol combined with collagen sponge;4 weeks as a treatment course, observation of 8 weeks, ulcer and other adverse reactions were recorded and compared.Results After 4 weeks of treatment, 17 cases (30.91%) with 30% or more reduction in foot ulcer area in treatment group,33 cases (60.00%) after 6 weeks treatment and the number of cases with 30% or more reduction in foot ulcer area,49 cases (89.09%) after 8 weeks of treatment, and the number of cases and the proportion of cases with 30% or more decrease of ulcer area in the control group after 4, 6 and 8 weeks were 12(20.00%), 22 (36.67%) and 39 (65.00%).There was no statistical difference between the two groups for the healing time of foot ulcer at three time points.After 8 weeks of treatment, the cure rate was 38.18% in salvia miltiorrhiza polyphenol combined with collagen sponge group. The effective rate was 50.91% and 45.00% for the two groups, the difference was not statistically significant; while the total effective rate was also significantly higher(89.09% vs.65.00%,P<0.01).Conclusion Salvia miltiorrhiza polyphenol combined with collagen sponge has a good effect on diabetic foot treatment, especially it can improve the cure rate, which is superior to the routine therapy.

Objective To compare the clinical efficacy of covered and uncovered metal stents in treatment of malignant biliary obstruction. Methods 123 cases of malignant biliary obstruction from May 2003 to May 2014. The survival time, the stent patency rates, the effective and biochemical indexes between the two groups were analyze and compared, follow-up period ended in March 2015. Results The level of bilirubin, transaminase, alkaline phosphatase (ALP) was declined after the covered and uncovered metal stents placed by ERCP, and the results of the two groups had no statistical significance (P > 0.05). Also there was no significant difference of the incidence of postoperative cholangitis and the cumulative survival rate between the two groups (P > 0.05). One year survival rates was related to tumor types (r = -1.55, P < 0.05). Conclusions Covered and uncovered metal stents for malignant biliary obstruction have no statistically significant about remission of liver function, stent occusion and the cumulative survival rate.

AIM To investigate effects of shenning on nephrotic rats. METHODS Adult male wistar rats were randomly devided into normal control group, pathological pattern group, high-dosage shenning group,middle-dosage group,low-dosage group,shenfukang group and prednisone group. Rats with membranous glomerulonephritis were induced by injection of cation bovine serum albumin into tail vein. The rats were treated respectively for 32 days by gavage. Urinary protein was observed once a week after 2 weeks. After 7weeks ,plasma albumin,renal histology,parameters of renal function and prostaglandins were examined. In addition TXB 2/6-Keto-PGF 1? in serum were also examined. RESULTS The urinary protein,serum cretinine level,BUN and TXB 2/6-Keto-PGF 1? can be reduced in shenning groups compared with pathological pattern group. Plasma albumin was significantly increased in high-dosage shenning group. CONCLUSION High-dosage and middle-dosage shenning can significantly decrease urinary protein in rats with membranous glomerulonephritis. High-dosage shenning can increase the plasma albumin level in rats with membranous glomerulonephritis.