The nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) has been shown to interact with a variety of cellular proteins and implicated in the regulation of cell growth, interferon resistance, and other cellular signaling pathways. Using the yeast-two hybrid method, we have isolated a clone that encodes a novel NS5A--associated binding protein: NS5ABP37. Reverse transcription polymerase chain reaction (RT-PCR) method was employed to amplify the full fragment,and the plasmid pGADT7-NS5ABP37 with the Saccharomyces cerevisiae vector pGADT7 was constructed. To prove the interaction, yeast cell Y187 transformed with pGADT7-NS5ABP37 was mated with yeast cell AH109 containing pGBKT7-NS5A to verify the interaction between the novel protein coded by the new gene NS5ABP37 and NS5A.

Protein-protein binding is the basis of virus and host cell interactions. With the application of technology of studying protein interactions, more knowledge of replication and pathogenesis of hepatitis C virus (HCV) was acquired. Non-structure protein 5B(NS5B) of HCV is a kind of viral protein, which plays an important role in replication of HCV. However, the effect of NS5B is not clear. To investigate the biological function of NS5B, we performed yeast two hybrid to look for proteins in hepatocytes interacting with NS5B. We constructed NS5B bait plasmid by cloning the gene of NS5B into pGBKT7, then transformed it into yeast AH109(a type). The transformed yeast was mated with yeast Y187(? type)containing liver cDNA library plasmid in 2?YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-?-gal for screening. Thirty-three colonies were selected and sequenced. Among them, two colonies were new genes with unknown function. The preliminary successful cloning of gene of protein interacting with NS5B paved the way for the study of the physiological function of NS5B and its associated protein.

Objective HCV NS3 protein plays an important role in disease caused by HCV. We investigate the gene expression of HCV NS3 in yeast for future study of the function of the protein. Methods PCR was performed to amplify the gene of HCV NS3 from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector. Thereafter, HCV NS3 gene was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, and recombinant pGBKT7∶NS3 was transformed into yeast AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. Results HCV NS3 gene was successfully cloned into pGBKT7. The results of SDS PAGE and Western blotting assay showed that the molecular weight of the expressed product was about 22000 Da and HCV NS3 protein was existed within yeast cells.Conclusions HCV NS3 was successfully expressed in yeast expression system.

Recent studies indicated that epigenetic abnormality is the important molecular mechanism in leukemia. The change of epigenetic modification occurs in most kinds of leukaemia. Based on the epigenetic modification, the therapies of leukemia with hypomethylating agents and histone deaeetylases inhibitors, which are different from traditional chemotherapy are applied in the treatment of leukemia at beginning. The aim of this article is to summarize the recent advances of DNA methylation and histone modification in leukaemia occurrence, treatment and prognosis.

To try to find out the possible use of histochemistry in the prenatal diagnosis in first trimester of pregnancy, 20 kinds of enzymes in chorionic villi from 6th to 9th week of gestation which are related to inborn errors of metabolism were investigated and graded according to their histochemical reactive intensity; the enzymes listed below: (1). ?-glucuronidase, (2). ?-galactosidase, (3). arylsulfatase, (4). ?-N-acetylglucosaminidase, (5). 3?-hydroxysteroid dehydrogenase, (6). NADH dehydrogenase, (7). adenosine triphosphatase, (8). alkaline phosphatase,(9). acid phosphatase, (10). creatine phosphokinase, (11). glutamic dehydrogenase, (12). aldolase, (13). xylitol dehydrogenase, (14). alcohol dehydrogenase, (15). xanthine oxidase, (16). catalase, (17). ornithine carbamoyl transferase, (18). lipase, (19). cholinesterase, (20). ?-glutamyl transpeptidase. The experimental results show that the last 6 kinds of enzymes present negative reactions. Since the his- tochemical reactions of the first 14 kinds of enzymes are positive, it is possible that they can be used for the prenatal diagnosis of inborn errors of metabolism caused by dificiency of corresponding enzymes. The results also show that compared with biochemical method, histochemical method has the advantages of requiring smaller amount of chorionic villi, being without contamination by maternal cells, simplicity and rapidity.

Objective To investigate the effect of escitalopram treatment on cognitive bias to the emotional facial information in patients with panic disorder. Methods 30 patients met CCMD-3 criteria for panic disorder were enrolled as research group and marched sexual and age 30 healthy persons enrolled as control group. Patients were treated with escitalopram for 8 weeks. All participants measured with dot-probe task of emotional facial information at base and after 8 weeks. RTs and attentional bias scores were compared respectively. Results After 8 weeks,HAMA scores (7. 81 ± 2. 52) in research group were lower than that of at base ( 17. 23 ± 3. 12) (P = 0.002). A repeated measure ANOVA revealed a significant probe site main effect (F(1,58) =4. 34, P = 0.031 ) , RTs of antarafacial site were longer than that of homonymy site. It revealed a significant probe site and group interaction(F(1,58) =16.15, P=0.000) ,a significant emotional facial information type and probe site interaction(F(1,58)=9.25, P =0.015) ,and a significant emotional facial information type × probe site× group interaction(F(1,58) =7. 31, P = 0. 002). LSD test showed that RTs of antarafacial site to fear facial information in research group were longer than that of homonymy site(P = 0.0009). RTs and attention bias scores of antarafacial site to fear facial information after 8 weeks in research group were lower than that of at base(P=0.032,0.008). Conclusion Patients with panic disorder have the cognitive bias to the fear facial emotional stimulus, and escitalopram treatment might improve the cognitive bias.

Aim To study the immunomodulatory activity of total flavonoids of Litsea Coreana Leveille (LCTF) on cyclophosphamide(CY)-induced immunosuppressive mice. Methods CY (50 mg?kg-1) was administered by intraperitoneal(ip)injection for 2 consecutive days to induce immunosuppressive model. Carbon clearance, quantitative hemolysis and DNFB-induced delayed-type hypersensitivity(DTH) were applied to assay effects of LCTF on nonspecific immunity, humoral immunity and cellular immunity. Results In carbon clearance test, the clearance index (K) and values of phagocytic index (?) were elevated by LCTF (200 and 400 mg?kg-1), indicating the phagocytosis of macrophages was enhanced in immunosuppressive mice.In quantitative hemolysis, productions of IgM and IgG in serum and hemolysin in splenocytes were enhanced in immunosuppressive mice by LCTF (100 and 200 mg?kg-1). LCTF (200 and 400 mg?kg-1) obviously increased DTH reactivity in immunosuppressive mice. LCTF not only increased percentages of T cells expressing CD4+ and CD8+,but also enhanced the ratio of the two subset of T lymphocyte,and LCTF (200 and 400 mg?kg-1) could also improve IL-2 production of spleen lymphocytes. Conclusion LCTF showed significant immunomodulatory property on immunosuppressive mice through specific and nonspecific immunity.

Objective To investigate the polyploid induction and identification of Nerviliae fordii for harvesting the polyploid plants. Methods The materials and methods for polyploid induction of Nerviliae fordii were screened separately by comparing the induction rates of rhizomes and bulbs under natural conditions and tissue culture environment, and by comparing soaking method with agar method. The effects of colchicine concentration ( 200, 300, 400, 300 mg/L), colchicine action time ( 7, 14, 21, 28 d), DMSO concentration ( 0, 10, 20, 40 mL/L) and KT concentration ( 0, 1.0, 2.0, 4.0 mg/L) on induction rate were observed by orthogonal design method. The polyploid induction in the treated plants was identified by morphology, cytology and chromosome methods. Results After the tissue culture rhizomes were treated with 300 mg/L colchicine, 10 mL/L DMSO, and 2.0 mg/L KT by agar method for 28 days, the polyploid induction rate arrived to 50%, showing better induction effect. The morphology of polyploid plants was characterized by giantism, and the leaf length, leaf width and plant height were respectively 152.17%, 158.67%and 60.90%of those of the diploid plants. The length, width and density of stoma of leaf epidermal cell as well as the number of chloroplast in the treated plants were 138.46%, 153.00%, 59.09% and 109.09% of those of the untreated plants. The results of chromosome identification showed that the amount of the tetraploid ( about 40) was 2 times of the diploid chromosome ( about 20) in the treated plants, proving that the achieved Nerviliae fordii was a tetraploid plant. Conclusion Polyploid plants of Nerviliae Fordii have been successfully obtained, which will supply evidence for improving species, richening seed-breeding resources, and selecting of improved seeds of Nerviliae fordii.

Chronic Disease ; Graft Rejection ; therapy ; Humans ; Integrative Medicine ; Kidney Diseases ; etiology ; therapy ; Kidney Transplantation ; adverse effects

This study investigated the expression of hemeoxygenase-1 (HO-1) in rats with acute lung rejection and its implication. A valid rat orthotopic left lung transplantation model (SD rat-->Wistar rat) was established by using an improved three-cuff anastomosis technique. The rats were divided into control group, CoPP (HO-1 inducer)-treated group and ZnPP (HO-1 inhibitor)- treated group. The severity of acute rejection was graded on the basis of the morphologic changes of the lung samples stained with HE. The expression of HO-1 protein in lung tissue was detected by using immunohistochemistry and Western blot, and HO-1 mRNA activity was assayed by RT-PCR. The results showed that the expression of HO-1 protein was significantly increased with the acute rejection grading in rats (P<0.01). As compared with control and ZnPP-treated groups, the severity of acute rejection was not alleviated and the grade not reduced significantly in CoPP-treated group (P>0.05). It was concluded that HO-1 protein might be involved in the pathological process of post-graft acute rejection. The expression of HO-1 protein was increased gradually with aggravation of acute rejection, and HO-1 protein might be used as an index to monitor acute rejection after lung transplantation.
Graft Rejection/*enzymology ; Heme Oxygenase (Decyclizing)/genetics ; Heme Oxygenase (Decyclizing)/*metabolism ; Lung Transplantation ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Rats, Sprague-Dawley ; Rats, Wistar