Acquired Immunodeficiency Syndrome ; China ; Cross-Sectional Studies ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male

Objective To explore the expressions of Survivin and VEGF and relationship between them in hepatocellular carcinoma(HCC).Methods The expressions of Survivin protein and VEGF protein in 50 HCC.30 cirrhosis and 10 normal tissues were assessed by immunohistochemical method.The expressions of Survivin mRNA and VEGF mRNA in 50 HCC,30 cirrhosis and 10 normal tissues were assessed by in situ hybridization.Results The expressions of Survivin and VEGF in cancer tissues,cirrhosis tissues,normal tissues weresignificantly different. The expression of Survivin in HCC tissues was stronger than that in cirrhosis,but the expreesion of VEGF in cirrho- sis was stronger than that in HCC tissues.Conclusion The expression of survivin.is closely associated with the ex- pression of VEGF in HCC and they take positive correlation.The abnormal expressions of Survivin and VEGF are closely associated with the development of HCC.They may play important roles in the development of HCC.

OBJECTIVETo explore the effect of ligustrazine injection (LI) on serum levels of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) in patients with bronchial asthma and determine the mechanism of action of LI in preventing and treating asthma.

METHODSSixty-eight patients with mild or moderate bronchial asthma were assigned to two groups equally according to their sequence number, odd or even. The conventional treatment was administered to both groups, and LI was given to the treatment group by ultrasonic spray inhalation twice a day but not to the control group. The therapeutic course for all was 2 weeks. Further, 30 healthy subjects who had no history of smoking were enrolled as the normal control group. The clinical condition scores, frequency of attacks and dosage of Terbutaline inhaled were scored and recorded on the first day of hospitalization (before treatment) and after treatment. At the same time, the indexes of lung function, including forced expiratory volume in one second (FEV1), forced expiratory volume in one second to forced vital capacity ratio (FEV1%) and the peak expiratory flow (PEF) were determined before treatment. The levels of IL-4 and IFN-gamma in peripheral blood were detected by ELISA before and after treatment, and compared with that of the healthy control group.

RESULTSAfter treatment, the clinical condition scores were found to be lower, indexes of lung function were elevated, but serum level of IL-4 and ratio of IL-4/IFN-gamma were reduced in both groups, showing significant differences as compared to those before treatment (P<0.05). However, the changes in all the indexes were more significant in the treatment group than in the control group, also showing statistical significance (P<0.05). No significant difference was revealed when IFN-gamma levels were compared before and after treatment in both groups, though a lowering trend could be seen, significant difference could not be found in the comparison of IFN-gamma levels between groups after treatment (P>0.05).

CONCLUSIONLI shows good clinical effect in treating bronchial asthma, and its mechanism might be related to the suppression of the synthesis of IL-4, thus leading to the inhibition of TH2 cell subset preponderant response and immune equilibrium regulation.

Adrenergic beta-Agonists ; administration & dosage ; therapeutic use ; Aged ; Aged, 80 and over ; Asthma ; blood ; drug therapy ; physiopathology ; Case-Control Studies ; Female ; Humans ; Injections ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; Pyrazines ; administration & dosage ; therapeutic use ; Respiratory Function Tests ; Terbutaline ; therapeutic use

OBJECTIVETo prepare and characterize monoclonal antibodies (mAbs) against the recombinant nucleocapsid (N) protein of 3 human coronaviruses SARS-CoV, 229E and OC43 and study the antigenic relationship between the 3 N proteins.

METHODSBALB/c mice were immunized with the recombinant N proteins of SARS-CoV, 229E and OC43 to obtain the mAbs by means of hybridoma. Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay (ELISA), Western blotting and indirect immunofluorescence assay. Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs.

RESULTSThe mAbs against the recombinant N proteins of SARS-CoV, 229E and OC43 were obtained, which reacted specifically with the corresponding viral N protein as shown by indirect ELISA, Western blotting and indirect immunofluorescence assay. No cross-reactivity was found between the 3 N proteins.

CONCLUSIONThe prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.

Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Coronavirus 229E, Human ; genetics ; immunology ; Coronavirus OC43, Human ; genetics ; immunology ; Cross Reactions ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; immunology ; SARS Virus ; genetics ; immunology

OBJECTIVETo observe the efficacy and safety of Jingui Shengqi Pill in treating partial androgen deficiency in aging males (PADAM), and to explore the new approach in improving the quality of life in PADAM patients.

METHODSForty patients with PADAM were treated with JSP, the efficacy was evaluated with international index of erectile function (IIEF) scoring, PADAM questionnaire scoring, hormone, prostatic specific antigen (PSA), etc., and the data before treatment were compared with those after treatment in the same group.

RESULTSAfter 3 months of treatment, PADAM scoring and IIEF scoring were all significantly improved. Symptoms regarding physical ability, vasomotion, and psychical and mental condition all got improved more markedly than symptoms regarding sexual hypofunction. The serum level of testosterone was 3.85 +/- 0.36 before treatment and 5.02 +/- 0.83 after treatment (P < 0.05); luteinizing hormone of 7.33 +/- 2.14 and 4.84 +/- 1.43 (P < 0.01), follicle-stimulating hormone of 10.22 +/- 4.48 and 6.47 +/- 3.28 (P < 0.01), respectively. The level of PSA failed to change significantly (1.94 +/- 0.55 and 2.06 +/- 0.47, P > 0.05).

CONCLUSIONJSP is effective and safe in treating PADAM, the mechanism of it is different from supplementing extrinsic androgen. It may have produced the effect by means of favorably regulating the condition of sex hormone to improve the balance of pituitary-sex gland axis, so it has more extensive clinical application.

Aged ; Androgens ; deficiency ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Testosterone ; blood

OBJECTIVETo clone and express avian influenza A virus [A/Hong Kong/482/97(H5N1)] H5 subtype hemagglutinin in baculovirus-insect cell expression system and investigate the antigenicity and bioactivity of the recombinant protein.

METHODSH5 gene of influenza A virus was amplified by PCR. The recombinant bacmid was obtained by cloning the gene to the donor plasmid of pFastBacHTB and transformed into DH10Bac competent cells. The recombinant baculovirus stock was prepared by transfecting the recombinant bacmid DNA into the insect cell line for protein expression after amplification. Immunofluorescene assay (IFA) and Western blotting were performed to identify the antigenicity of the recombinant protein, and hemagglutination assay was used to identify its bioactivity.

RESULTSThe recombinant his-H5 protein was expressed in the insect cells with a relative molecular mass of 64,000, which showed erythrocyte-agglutinating activities with the red blood cells of guinea pig. Western blotting and IFA demonstrated that the recombinant his-H5 could be recognized and bound by standard anti-H5 sera.

CONCLUSIONThe recombinant his-H5 with a post-translation modification is successfully obtained in insect cells, which may provide a potential source for further study of the antigen's biological function and for production of the subunit vaccine or monoclonal antibodies.

Animals ; Baculoviridae ; genetics ; Cell Line ; Erythrocytes ; cytology ; immunology ; Genetic Vectors ; genetics ; Guinea Pigs ; Hemagglutination Tests ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; immunology ; Spodoptera ; Transfection

Objective To study the morbidity of cough variant asthma (CVA) among patients with chronic cough syndrome and its relative risk factors. Methods Patients were recruited with detailed history on their illness. Data were collected on physical examination, chest X-ray, eosinophil cell counts, pulmonary ventilation with histamine stimulating test and bronchi dilation test. According to available data, diagnosis of CVA was confirmed and the relative factors Questionnaire form was completed for each patient. Results Among 473 patients with chronic cough, 95 (44 male and 51 female) were confirmed to be CVA (20.08%). Analysis of the relative factors suggested that CVA was associated with multiple factors. Morbidity of CVA was associated with season, personal histories on allergy and family history on asthma, CVA could be induced by upper respiratory tract infection, inhale of oil vapor, acrimony air, over-burdened physical exercises etc. Conclusion For patients with chronic cough symptom, clear diagnosis of CVA, avoid of passable risk factors and timely medical intervention when necessary, would be helpful in controlling clinical courses and improving the prognosis of the disease.

Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs used in experiment were divided into three groups:control group,mannitol group and hypertonic glucose group.HPMCs in control group received no treatment,and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours,respectively.Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB,E-cadherin,α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),Cyclin D1,cyclin dependent kinase inhibitor 4 (CDK4),protein 27 (p27)and proliferating cell nuclear antigen (PCNA).Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA,CTGF,Cyclin D1,CDK4,p27 and PCNA,and flow cytometry was used to test the cell cycle.Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs.Real-time PCR was employed to detect the mRNA expression of E-cadherin,α-SMA and CTGF,Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA and CTGF,and flow cytometry was used to test the cell cycle.Results It is revealed by Real-time PCR,Western blotting and flow cytometry that the expressions increased of LncRNA-ATB,α-SMA,CTGF,Cyclin D1,CDK4 and PCNA induced by hypertonic glucose,and decreased of E-cadherin and p27 (P＜0.05).Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation,while down-regulation alleviated HPMCs phenotypic transition and proliferation.Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.

OBJECTIVETo prepare and characterize monoclonal antibodies (mAb) and polyclonal antibodies against nucleocapsid (N) protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) and to establish antibodies-based sandwich ELISA for detecting N protein of SARS-CoV, which might apply to early diagnosis of patients with SARS-CoV infection.

METHODSBALB/c mice were immunized with purified recombinant N protein of SARS-CoV for producing mAbs, and New Zealand white rabbits were immunized for producing polyclonal antibodies. The identification of antibodies was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. Capturing and detecting antibodies were selected by pairing the mAbs and polyclonal antibodies one by one and an antibodies-based sandwich antigen capture ELISA was used for detecting N antigen of SARS-CoV.

RESULTSNine mAbs and hyperimmune rabbit polyclonal antibodies, specifically against SARS-CoV nucleocapsid protein were obtained. Using paired ELISA assay, three mAbs N1E8, N8E1 and N10E4 were selected as capturing antibody and rabbit polyclonal antibodies as detecting antibody then triple antibodies-based sandwich ELISA was established following horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G. The recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml with this assay. When tested with 420 serum specimens from serologically confirmed SARS patients, the positive rates of serum N protein were 90.1%, 23% and 0%, in which sera collected from 1 to 10 days, 11 to 20 days and beyond 21 days respectively after the onset of symptoms. The specificity of the assay was 99.86% in 715 control serum specimens. There was no cross-reaction with other respiratory viruses and coronaviruses.

CONCLUSIONSpecific and high affinity mAbs and rabbit polyclonal antibodies were obtained. By paired and optimized sandwich ELISA, a sensitive and specific antigen capture ELISA was established for detecting N antigen of SARS-CoV, which might apply to early diagnosis, source tracing and epidemiological studies of SARS.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid ; immunology ; Rabbits ; SARS Virus ; immunology ; isolation & purification ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; virology

OBJECTIVETo produce neutralizing antibodies against envelope protein domain III (EDIII) of dengue virus serotype I (DENV-1) and evaluate the nonstructural protein 1 (NS1) antigen capture enzyme-linked immunosorbent assay (ELISA) for identification of antibody neutralizing abilities.

METHODSFive BALB/c mice and one New Zealand Rabbit were immunized with recombinant EDIII protein of DENV-1 for the production of hybridomas and hyperimmune sera. Indirect ELISA, immunofluorescence assay (IFA) and Western Blot analyses were applied to identify specificity of antibodies. Comparing to plaque reduction neutralization test (PRNT), the new established DENV-1 specific NS1 antigen capture ELISA was used for detecting the neutralizing abilities of these antibody.

RESULTSFour strains of monoclonal antibodies (mAbs) named 1A1, 1B3, 3D3 and 9D6 and one hyperimmune serum of rabbit were obtained, all of which were approved to have neutralizing abilities to DENV-1 with the PRNT titer of 1:1024, 1:512, 1:256, 1:4096 and 1:4096. MAb 3D3 with the lowest neutralization titer in PRNT had not shown neutralizing ability to DENV-1 in NS1 antigen capture ELISA, while MAbs 1A1, 1B3 and 9D6 and the rabbit hyperimmune serum could protect the C6/36 from being infected by DENV-1 with the neutralization titer of 1:32, 1:32, 1:128 and 1:128 in this assay.

CONCLUSIONNS1 antigen capture ELISA could be used to identify antibody neutralizing abilities to DENV, it was a faster and more convenient way to screen antibodies with high neutralization titer and might also be used as one of the methods to evaluate the effects of vaccines.

Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Dengue Virus ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Mice ; Mice, Inbred BALB C ; Neutralization Tests ; Rabbits ; Viral Envelope Proteins ; immunology ; Viral Nonstructural Proteins ; immunology