OBJECTIVETo detect the in-vitro effects of arbidol hydrochloride against 2009 new influenza virus A (H1N1).

METHODSThe activity of arbidol hydrochloride against 2009 new influenza virus A (H1N1) was determined in MDCK cell cultures. Hemagglutination assay, observation of cytopathic effects, RT-PCR and quantitative RT-PCR tests were performed for determination of virus titers. Inhibition concentration 50% and cytotoxic concentration 50% were calculated with Chou's Menu of Dose-Effect Program.

RESULTSArbidol hydrochloride showed low cytotoxicity (cytotoxic concentration 50%>100 μmol/L)and significant anti-2009 new influenza virus A (H1N1) activity in cell cultures. Inhibition concentration 50% were (5.5 ± 0.9), (3.4 ± 0.8), and (1.5 ± 0.2) μmol/L in hemagglutination assay, cytopathic effect test, and quantitative RT-PCR assay, respectively.

CONCLUSIONArbidol has low cytotoxicity and high anti-virus activity and can effectively trigger the activities of interferon and immune response, and therefore can be a valuable anti-influenza virus drug.

Animals ; Antiviral Agents ; pharmacology ; Dogs ; Indoles ; pharmacology ; Influenza A Virus, H1N1 Subtype ; drug effects ; Madin Darby Canine Kidney Cells ; Virus Replication ; drug effects

Humans ; In Situ Hybridization, Fluorescence ; Membrane Proteins ; analysis ; NF-kappa B ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Tissue Array Analysis ; methods ; Tumor Cells, Cultured

Objective To observe the changes of antithrombin activity(AT) and D-dimer in acute leukemia(AL)children complicated with disseminated intravascular coagulation(DIC) and to explore the changes of blood coagulation and fibrinolysis function.Methods Twenty-seven AL children without DIC were selected as AL group and 25 childern complicated with DIC were selected as observe group,36 health children were as control group.Plasma level of AT,D-dimer,fibrinogen,activated partial thromboplastin time and prothrombin time were tested by color substrate,immuno-latex turbidimetry,and coagulation method.And the rusults of AL group were compared with observe group and control group by SPSS 10.0 software.Results PT was significantly prolonged and the D-dimer in AL group and observation group were significantly higher than those in control group(Pa

In the two decades since AZT was first approved for clinical use in 1987, 24 additional antiretroviral agents have been approved. They include 7 nucleoside analogs, a nucleotide analog and 4 non-nucleoside reverse transcriptase inhibitors, 10 protease inhibitors, 2 entry inhibitors and an integrase inhibitor. More than 20 investigational agents are currently being studied in clinical trials. Highly active antiretroviral therapy (HAART), which involves a combination of anti-HIV-1 drugs, is extremely effective in suppressing HIV-1 replication and increasing CD4+ number and results in substantial reductions in HIV-1-related morbidity and mortality. In last 20 years, much has been learned about resistance to antiretroviral drugs, drug interactions and metabolic complications of antiviral drug use. Drugs are now selected on the basis of resistance tests and on the risk of specific drug complications in individual patients. As a result, decisions about the therapy of HIV/AIDS have become personalized and are made on a patient-by-patient basis. With appropriate medical management, a person with HIV-1 now has the possibility of a nearly normal life expectancy.
Anti-HIV Agents ; adverse effects ; pharmacology ; therapeutic use ; Antiretroviral Therapy, Highly Active ; Cyclohexanes ; chemistry ; pharmacology ; therapeutic use ; Drug Resistance, Viral ; HIV Envelope Protein gp41 ; chemistry ; therapeutic use ; HIV Fusion Inhibitors ; chemistry ; pharmacology ; therapeutic use ; HIV Infections ; drug therapy ; HIV Integrase Inhibitors ; chemistry ; pharmacology ; therapeutic use ; HIV Protease Inhibitors ; chemistry ; pharmacology ; therapeutic use ; HIV Reverse Transcriptase ; chemistry ; pharmacology ; therapeutic use ; HIV-1 ; drug effects ; physiology ; Humans ; Molecular Structure ; Peptide Fragments ; chemistry ; therapeutic use ; Pyrrolidinones ; chemistry ; pharmacology ; therapeutic use ; Raltegravir Potassium ; Saquinavir ; chemistry ; pharmacology ; therapeutic use ; Triazoles ; chemistry ; pharmacology ; therapeutic use ; Virus Replication ; drug effects ; Zidovudine ; chemistry ; pharmacology ; therapeutic use

OBJECTIVETo investigate the distribution of hepatitis B virus (HBV) genotype in Guizhou and to study the relationship between the genotype and the progression of liver disease.

METHODS786 patients with chronic HBV infection, from 4 cities of Guizhou, including 346 asymptomatic carriers (ASC), 313 chronic hepatitis (CH), 77 liver cirrhosis (LC), 50 hepatocellular carcinoma (HCC) were examined. HBV genotype was determined by restriction fragment length polymorphism analysis and the subtypes were determined by direct sequencing of PCR product in 94 patients with HBV B genotype, the relationship between HBV genotype and the progression of liver disease was studied by multifactor analysis such as HBeAg positivity, HBV DNA load and ALT level.

RESULTSOf the 786 patients, 7 (0.89%), 497 (63.23%), 275 (34.99%), and 7 (0.89%) belonged to genotype A, B, C, D, respectively. There was statistically significant difference in the distribution of genotype B among Kaili (96.04%), Zunyi (78.79%), Duyun (64.52%) and Guiyang (53.14%) (P< 0.01). Genotype C was more prevalent in Guiyang than in other three cities (P < 0.01, or P < 0.05). Out of 94 genotypes B, 93 (98.94%) belonged to subtype Ba, only one was subtype Bj. There were statistically significant difference in the distribution of genotype B and C among various stage of liver disease (P < 0.05 or P < 0.01). Genotype B showed a gradual decrease from ASC, CH, LC to the HCC group while in contrast, genotype C showed a gradual increase in the same order. The ALT levels and the mean age were significantly higher and older in patients with genotype C than those in genotype B (P < 0.01 or 0.05). The HBeAg positivity was significantly lower in genotype C than that in genotype B (P < 0.025).

CONCLUSIONData showed that there were genotype A, B, C and D existing in Guizhou. Genotype B was the major one but genotype C was more commonly seen. In genotype B, subtype Ba appeared to be predominant. The geographic distribution of genotype B and C were different in some cities of Guizhou. Compared to genotype B, genotype C was associated with the development of more severe liver damage.

Carcinoma, Hepatocellular ; pathology ; virology ; DNA, Viral ; analysis ; Disease Progression ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; genetics ; pathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; virology ; Liver Neoplasms ; pathology ; virology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

The expression of the vgb gene in vivo could improve the fermentation density and then contribute the extracellular secretion of the product of bxn gene. Constructed the recombination plasmid pPIC9K-vgbbxn and transformed into Pichia pastoris GS115. The results of PCR and SDS-PAGE indicate that the vgb gene and bxn gene had integrated into the genome of Pichia pastoris GS115 and expressed in efficient level. Also, the protein activity of their products had been verified respectively. Shake flask fermentation experiments showed that the presence of VHb in yeast Pichia pastoris efficiently enhanced cell growth and secretive expression of bxn gene under hypoxic habitats.
Aminohydrolases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; physiology ; Electrophoresis, Polyacrylamide Gel ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Truncated Hemoglobins ; genetics ; physiology

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo study the efficacy, safety and reliability of colonic sac duct for first-stage repair of colorectal anastomotic leakage.

METHODSAn animal model of colon anastomotic leakage was established in 30 Tibet miniature pigs, which were randomly divided into treatment group and control group (n=15). Colon anastomotic leakage in the treatment group was repaired using the colonic sac duct, while the control group received conventional surgical repair. At 7, 14, and 21 days after the surgery, the healing of the anastomotic leakage was evaluated by examining the bursting pressure, tissue microvessel density and hydroxyproline content at the anastomosis.

RESULTSUsing the colonic sac duct, the anastomotic leakage was successfully repaired without death of the pigs or the occurrence of intestinal stenosis or necrosis. At 7 and 14 days after the surgery, the bursting pressure, hydroxyproline contents, and microvessel density in the treatment groups were higher than those in the control group, but such difference was not found at 21 days.

CONCLUSIONColonic sac duct allows effective repair of colon anastomotic leakage, and is especially useful for leakage lasting for 48-72 h complicated by severe abdominal infection.

Anastomosis, Surgical ; adverse effects ; Anastomotic Leak ; etiology ; surgery ; Animals ; Colon ; surgery ; Female ; Male ; Rectum ; surgery ; Swine ; Swine, Miniature

OBJECTIVETo investigate the epidemic state of arboviruses in the downstream area of Lancang river in Yunnan province.

METHODSMosquitoes were collected from Lancang river downstream area (including Lancang county and Simao city) during summer in 1998 and stored in liquid nitrogen after classification. The mosquito pools were homogenized and centrifuged, then the supernatant was inoculated into C6/36 cells for virus isolation. New isolates were identified by neutralization test(NT), ELISA, immunofluorescence assay(IFA) and polyacrylamid gel electrophoresis(PAGE).

RESULTSTotally 22 isolates of arbovirus were obtained from 233 mosquito pools by inoculation of C6/36 cells and positive rate of the isolation was 9.4%. Ten strains were resistant to both ether and 5 prime-IDU. So they were non-enveloped double-stranded (ds) RNA virus. Twelve segmented RNAs were shown by PAGE and PAGE profiles from the ten strains were 6-6 with minor variation. The isolates can be neutralized by immunized mouse ascites fluid of BJ95-75 strains of coltivirus by NT, and reacted with monoclonal antibody against BJ95-75 by ELISA. These viruses were identified as coltivirus. Nine isolates were sensitive to ether and resistant to 5 prime-IDU. So they were non-enveloped RNA viruses. PAGE showed 10 segmented RNA, and they were identified to be orbiviruses. Three isolations were sensitive to ether. One of them can be neutralized with JEV A2 strain antibody by NT and was positive to the homologous antibody by IFA. It was identified being strain of JE virus. One strain(YN92-4) can be reacted with anti-bunyavirus group specific immune ascites fluid by both IFA and ElISA, but reacted neither with anti-alpha virus group, nor with anti-flavivirus group JE virus ascites fluid. The virions are spherical and about 87 nm in diameter with surface projections by negative staining. Conclusion Twenty-two isolates have been obtained from wild caught-mosquitoes of Lancang river down-stream area in Yunnan province. Among them ten, nine, one and one were identified as coltivirus, orbivirus, JE virus and bunyavirus, respectively. One is under identification. This is the first report on bunyavirus isolated from mosquitoes in China.

Animals ; Arboviruses ; classification ; isolation & purification ; China ; Coltivirus ; isolation & purification ; Culicidae ; virology ; Encephalitis Virus, Japanese ; isolation & purification ; Insect Vectors ; virology ; Orbivirus ; isolation & purification ; Orthobunyavirus ; isolation & purification

OBJECTIVETo determine the clinical results of selected CD34(+) cell autologous transplantation in advanced malignant tumors.

METHODSAfter pretreatment, fifteen patients aged 12 - 70 (49.5) years with various Stage III or IV malignant tumors were given the sorted CD34(+) cells collected by magnetic-activated cell sorting (Clini MACS, Milteny Biotech, Germany).

RESULTSPeripheral blood progenitor cells (PBPC) from the patients were mobilized by chemotherapy and G-CSF 5 micro g/kg per day. CD34(+) cells gave 2.0 - 5 log depletion after cell sorting, with a median yield of CD34(+) selected cells of 2.4 (0.15 - 12.03) x 10(6)/kg. It gave a median recovery of 64 (52 - 81.4)% and median purity of 98.2 (83.2 - 99.7)%. The median time of neutrophil recovery > 1.0 x 10(9)/L and platelet recovery > 20 x 10(9)/L post-transplantation were 14 (8 - 26) days and 13 (11 - 35) days, respectively. On follow-up of 2 - 33 (11) months, the event-free survival rate was 53.3% (8/15) and the overall survival rate was 66.7% (10/15).

CONCLUSIONTransplantation of autologous selected PBPC CD34(+) cells gives prompt and stable engraftment. Selected CD34(+) cell transplantation, being a safe approach, may improve the clinical outcome even in patients with advanced malignant tumors.

Adolescent ; Adult ; Aged ; Antigens, CD34 ; analysis ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Neoplasms ; mortality ; therapy ; Survival Rate ; Transplantation, Autologous