Objective To construcr a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein. Methods The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by RT-PCR from peripheral blood Lymphocytes mRNA of patients with positive serum hepatitis B virus core protein antibody and randomly combined through a DNA linker encoding the peptide (Gly 4Ser) 3 to construct single-chain variable fragments (ScFv)gene. In the presence of helper phage M13KO7, the ScFv were displayed on the surface of recombinant phages. Results Repertoire antibody library was obtained with 10 6 clonies resistant to ampicillin and kanamycin. Conclusions A typical phage display library of human single-chain Fv antibodies has been constructed Using RT-polymerase chain reaction (RT-PCR) and phage display technique, human single-chain Fv antibodies can be screened from this library.

OBJECTIVE:To investigate the inhibitory effect of the total flavoniods of Ampelopsis grossedentata on human hepatoma carcinoma cells(HCC) in vitro.METHODS:The inhibitory effect of the total flavoniods of Ampelopsis grossedentata extract on human hepatoma carcinoma BEL7404 cells in vitro was analyzed by MTT,cell growth curve method and colony formation method,respectively.RESULTS:The total flavonoids of Ampelopsis grossedentata significantly inhibited the growth of BEL 7404 cells in vitro.MTT assay showed that the IC50 was 28.59 ?g?mL-1;the cell growth curve measurement showed that the total flavonoids of Ampelopsis grossedentata significantly inhibited the growth of BEL 7404 cells;the colony formation assay showed that the IC50 was 100% at a drug concentration of above 22.22 ?g?mL-1 and the IC50 was 58.05% at a drug concentration of 14.81 ?g?mL-1.CONCLUSION:The total flavonoids of Ampelopsis grossedentata have a remarkable inhibitory effects on the growth of BEL 7404 in vitro.

Obiective:To investigate the effect of interleukin-6(IL-6)on apolipoprotein(apo)M expression in a human hepatoblastoma cell line(HepG2).Methods:HepG2 cells were cultured and incubated with different concentrations of IL-6 (0,1.25,2.5,5,10,20,40 and 80 μg/L)for 24 hours.After the incubations,total RNAs were extracted and applied to reverse transcript PCR and real-time quantitative PCR to detect the expression levels of apoM and apoA-I.The effect of IL-1α on apoM expression was also determined.Results:IL-6 significantly inhibited the expression of apoM(F=10.778,P ＜ 0.01).Whereas IL-6 did not influence the expression of apoA-I(F=2.004,P ＞ 0.05).IL-1α(0,1.25,2.5,5,10,20,40 and 80 μg/L)did not decrease the expression of apoM(F=2.038,P ＞ 0.05).Cooclusion:IL-6 inhibits apoM mRNA transcription,which may contribute to the pathogenesis of abnormal lipid metabolism and macrovascular complications in type 2 diabetes.

Objective To analyze the genetic characteristics of measles virus strains causing two outbreaks in Guizhou province from November 2014 to March 2015. Methods Throat swab samples collect-ed from measles cases in two outbreaks were inoculated into Vero/SLAM cells. Viral RNAs were extracted from positive cultures. Nucleoprotein genes were amplified by using reverse transcription-polymerase chain reaction ( RT-PCR) and the PCR products were sequenced and analyzed. Results Eleven strains of wild-type measles virus were isolated from the two measles outbreaks and all of them belonged to H1a sub-geno-type. Phylogenetic analysis showed that those strains were clustered into two distinct branches. Differences in nucleotide and amino acid genetic distances between the 11 strains of measles virus and the WHO reference strain of H1a sub-genotype (Chin9322) were 1. 1%-1. 6% and 0. 7%-3. 4%, respectively. Compared with the reference strain Chin9322 and Guizhou epidemic strains in recent years, six strains showed amino acid sequence mutations in 47 ( G to S) , 82 ( S to G) and 122 ( R to K) sites and two strains had a mutation in 98 ( P-L) site. Conclusion H1a sub-genotype measles virus was the predominant pathogen causing two measles outbreaks in Guizhou province during 2014 to 2015. Moreover, it was also a predominant sub-geno-type circulating in China and Guizhou province. Different measles virus strains of H1a sub-genotype contin-ued to be prevalent in Guizhou province. This study provided some scientific data for the control and elimina-tion of measles in Guizhou province.

Objective To evaluate the efficacy of different doses of dexmedetomidine mixed with ropivacaine for brachial plexus block.Methods One hundred and twenty ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 20-60 yr,weighing 40-70 kg,scheduled for upper extremity surgery under brachial plexus block,were randomly divided into 6 groups (n =20 each):ropivacaine group (group R) and different doses of dexmedetomidine groups (groups RD1-5).Brachial plexus block was performed using the interscalene approach.In group R,interscalene brachial plexus block was performed using 0.5 ％ ropivacaine 25 ml (single injection).In groups RD1.5,interscalene brachial plexus block was induced with a mixture (25 ml) of dexmedetomidine 0.25,0.50,0.75,1.00,1.25μg/kg and 0.5 ％ ropivacaine,respectively.The onset time and duration of sensory and motor block were recorded.The adverse effects such as adverse cardiovascular events,excessive sedation,and pneumothorax were also recorded.Results Compared with group R,the onset time was significantly shortened,and the duration of block was prolonged (P ＜ 0.05).Compared with groups RD1 and RD2,the onset time was significantly shortened,and the duration of block was prolonged in groups RD3 5 (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups RD1 and RD2 or among groups RD3,RD4 and RD5 (P ＞ 0.05).Some patients developed bradycardia,hypotension or excessive sedation in groups RD4 and RD5,while no adverse effects were observed in the other groups.Conclusion Dexmedetomidine 0.75 μg/kg mixed with 0.5 ％ ropivacaine 25 ml can be safely and effectively used for brachial plexus block in patients.

Objective To investigate whether the combination of hydroxyapatite/tri-calcium phosphase (HA/TCP) and distraction osteogenesis technique would greatly reduce the time for the treatment of bone defects and enhance bone consolidation.Methods Osteotomy was made in the left tibia of all the 36 rabbits (age 20-24 weeks,body weight 2.2-2.8 kg),and a 1.0 cm length of the tibial shaft was removed below the tibiofibular junction.All rabbits were randomly divided into 3 groups,12 per group.Group A:The 1.0 cm defect gap was immediately reduced with the tibia shortened for 1.0 cm.Group B:The 1.0 cm defect gap was immediately filled with 1.0 cm restorable porous HA/TCP cylindrical block.Group C:The 1.0 cm defect gap was immediately reduced with the tibia shortened for 0.5 cm,and the remaining 0.5 cm defect gap was filled with 0.5 cm restorable porous HA/TCP cylindrical block.Then all the tibia was fixed with unilateral lengthener.For Group A and Group C,the lengthening started on the seventh day postoperatively,and lasted for 10 days and 5 days respectively.Group B didn't perform lengthening.All the rabbits wore terminated on the 37th days postoperatively.Serial radiographs were taken on the day of surgery,12,17,27 and 37 d postoperatively.The excised bone specimens were subject to micro-CT,mechanical testing,and histological examinations.Results Compared with the other two groups,the results in Group C were better in the terms of bone mineral content 454.44 ±89.98 mg,tissue mineral content 454.40±89.97 mg,maximum torque,maturity of regenerate bone,and the speed of bone consolidation and remodeling.Complete bone healing was achieved in the Group C within 37 days,but not in the Group A and B within 37 days.Conclusion The combination of HA/TCP and distraction osteogenesis technique can reduce the treatment time and promote bone consolidation compared with single treatment.

Acupuncture Points ; Biological Evolution ; Humans ; Terminology as Topic

Adolescent ; Body Height ; Body Weight ; Child ; Female ; Growth and Development ; Humans ; Male ; Papilloma ; surgery ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; adverse effects

Objective To investigate the clinical efficacy of sequential plasma perfusion of low dose plasma exchange in the pa-tients with acute on chronic liver failure (ACLF).Methods The patients with HBV related ACLF in the infection department of the Affiliated Hospital of Luzhou Medical College were divided into the plasma exchange therapy group (group A,n=114)and the low dose plasma exchange sequential plasma perfusion therapy group (group B,n=144).The group A was treated only by plasma exchange of fresh frozen plasma 2 400 mL;while the group B adopted the sequential plasma perfusion of plasma exchange,with fresh frozen plasma dosage of 1 400 mL.The changes of the liver and renal function,coagulation function and electrolyte indicators were observed in the two groups.The differences in the clinical effects and adverse reactions were compared between the two groups.Results The serum levels of ALT,TBIL and INR after treatment in the two groups were significant decreased than before treatment(P<0.05);ALB and PTA were significantly improved compared with before treatment(P<0.05);there were no signif-icant differences in serum levels of CRE,K+ and Na+ between before and after treatment.The changes of various indexes before and after treatment had no statistical differences between the two groups.The total effective rate and the mortality at 24 weeks in the group B was 66.66% and 47.22% respectively,which had no statistical differences compared with the group A.The total oc-currence rate of adverse reactions in the group B was 23.75%,which was significant lower than 35.26% in the group A with statis-tical difference (P=0.011).Conclusion Sequential plasma perfusion of plasma exchange has better clinical effect and few adverse reactions for treating the patients with ACLF,which can reduce the plasma dosage significantly.

BACKGROUND:Tol-like receptor 4 (TLR4) and its ligand, lipopolysaccharid, are closely associated with the occurrence and development of periodontitis. Meanwhile, the immunological properties of periodontal ligament stem cells (PDLSCs) play an important role in the reconstruction of periodontal tissue and cel-based therapy of periodontitis. However, the effect of TLR4 and lipopolysaccharid on the immunological properties of PDLSCs remains unclear. OBJECTIVE:To investigate the effect of TLR4 on the immunological characteristics of human PDLSCs. METHODS:PDLSCs were isolated by enzyme digestion method as previously reported, and were cultured in the medium containing 10 mg/L lipopolysaccharid, the ligand of TLR4 for 3 days. Using un-treated PDLSCs as controls, we then investigated whether lipopolysaccharid-treated PDLSCs could cause the proliferation of al ogeneic T lymphocytes as wel as the effect of lipopolysaccharid-treated PDLSCs on the mixed lymphocytes reaction and proliferation of lymphocytes induced by phytohemagglutinin. PDLSCs, peripheral blood mononuclear cells and phytohemagglutinin were co-cultured, and the concentration of prostaglandin E2 in the culture supernatant was examined. Then we added indomethacin, which is the inhibitor of prostaglandin E2, into the co-culture system of PDLSCs, peripheral blood mononuclear cells and phytohemagglutinin, and tested the proliferation of lymphocytes. RESULTS AND CONCLUSION:Lipopolysaccharid-treated PDLSCs did not lead to the proliferation of al ogeneic T lymphocytes just as un-treated PDLSCs, and could suppress the mixed lymphocytes reaction and proliferation of phytohemagglutinin-induced lymphocytes. However, the inhibitory ability of lipopolysaccharid-treated PDLSCs was significantly lower than that of un-treated PDLSCs. The levels of prostaglandin E2 were significantly elevated in the co-culture of PDLSCs, peripheral blood mononuclear cells and phytohemagglutinin. After adding of indomethacin, the PDLSCs-suppressed proliferation of lymphocytes restored to normal levels. Lipopolysaccharid weakened the immunosuppressive capacity of PDLSCs, which may be due to the decreasing secretion of prostaglandin E2.