Adolescent ; Cholesteatoma ; complications ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans ; Male ; Temporal Bone

Objective To investigate the clinical spectrum of respiratory viruses in infants and young children with acute lower respiratory infection(ALRI) in Chongqing area from 2003-2007.And to assess the clinical diagnostic value of virus detection in nasopharyngeal secretions(NPS) and serum viral antibody detection for ALRI.Methods Cases of 2 529 specimens of NPS in hospitalized children with ALRI from Apr.2003 to Oct.2007 were taken for detecting 7 common respiratory virus antigens by immunofluorescence assay including respiratory syncytial virus (RSV),adenovirus(ADV),influenza A(IA),influenza B (IB),parainfluenza virus1-3 (PIV1,PIV2,PIV3).Fifty-five thousand eight hundred and eighty-seven samples were tested for ADV-IgM by ELISA.Among those,45 159 cases were further tested for RSV-IgM by ELISA.Results Respiratory virus pathogens were detected in 778 samples out of 2 529(30.76%) including RSV positive in 668 samples (85.86%),PIV3 positive in 75 samples (9.64%),IA positive in 22 samples (2.57%),ADV positive in 15 samples ( 1.93%),only 1 sample ( 0.13%) positive for both PIV1 and RSV. And the positive rate of RSV-IgM was 0.9%-15.2%,and the positive rate for ADV-IgM was about 0.6%-10.6%.RSV infection occured mainly in winter and spring.Conclusions Respiratory virus is the most common pathogen in children with ALRI during the survey period in Chongqing area,especially for RSV infection.The pattern of RSV circulation varied every year with seasonality.It is suggest that this year is peak one for RSV infection from the monthly positive results,especially in Feburary(50%) in 2007.But the infection rate of PIV3,IA,ADV and PIV1 are lower,particularly IB and PIV2 infection have not been seen for the last 5 years.It is fast and accurate to detect RSV antigen and suit to clinical diagnosis by using immunofluorescence assay than other antibody detection.

The double-antibody-sandwich enzyme-linked immunoadsorbent assay (ELISA) for detection of rLysostaphin in humans had been developed and established through this study. rLysostaphin of high purity ( > 95 % ) produced in Shanghai Hi-Tech United Bio-Technological Research & Development Co., Ltd (SHUBRD) was used to produce a rabbit anti-rLysostaphin polyclonal antibody. The standard curve of rLysostaphin polyclonal antibody that was constructed showed that the lowest range of detection was found at 0. 98 ng of rLysostaphin/mL, and the curve exhibited linearity preferably from 0. 98 to 500 ng of rLysostaphin/mL. When three serum samples of the same batch were assayed for 6 replicates, and more 3 samples from different batches for 6 replicates, the average intra-assay and inter-assay coefficient variances ( CV) were 6. 4% and 6. 5%, respectively. The relative recovery rate was 98.6% when quantitative standard antigens were added to the serum. The present method for detection of rLysostaphin in serum is specific, highly sensitive, highly precise, and exhibited a low CV and will be helpful in the further study of rLysostaphin pharmacokinetics and holds promise in clinical applications.
Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Enzyme Stability ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immune Sera ; immunology ; Lysostaphin ; blood ; immunology ; metabolism ; Male ; Rabbits ; Recombinant Proteins ; immunology ; metabolism ; Reproducibility of Results ; Temperature

OBJECTIVETo investigate the effects of human cytomegalovirus (HCMV) on the proliferation of duct epithelial cells of human salivary gland (HSG).

METHODSThe expression of proliferating cell nuclear antigen (PCNA) and p53 were studied in 11 cases of parotid cytomegalic inclusive disease (PCID) using immunohistochemical staining method. The effects of human cytomegalovirus (HCMV) on the proliferation of HSG were investigated by MTT method in vitro. The expression of PCNA in HSG infected by HCMV was examined using immunocytochemical staining and Western blotting.

RESULTSPCNA was expressed weakly in most of megalic inclusion cells which were positive for HCMV, while all the megalic inclusion cells were p53 negative in all 11 cases of PCID. HCMV inhibited proliferation of HSG in vitro in a time dependent and dose dependent manner. Down-regulation of PCNA was shown in infected cells.

CONCLUSIONHCMV inhibits proliferation of HSG and down-regulation of PCNA may be an expression of the inhibition.

Cell Division ; Cells, Cultured ; Cytomegalovirus ; genetics ; pathogenicity ; physiology ; Cytomegalovirus Infections ; genetics ; pathology ; Down-Regulation ; Epithelial Cells ; pathology ; Female ; Humans ; Male ; Parotid Gland ; pathology ; virology ; Proliferating Cell Nuclear Antigen ; analysis ; Salivary Ducts ; pathology ; virology ; Tumor Suppressor Protein p53 ; analysis

OBJECTIVETo explore the correlations of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) with the clinicopathological characteristics, prognostic events, and survival outcomes in esophageal cancer (EC) patients.

METHODSThe PubMed, Web of Science, Embase database and Cochrane database were searched for studies reporting the outcomes of interest. The studies were selected according to established inclusion/exclusion criteria. Meta-analysis of the studies was performed using Review Manager 5.3 and Stata12.0 software with the odds ratio (OR), risk ratio (RR) , hazard ratio (HR) , and 95% confidence interval (95% CI) as the effect indexes.

RESULTSNineteen studies involving a total of 1766 patients were included in the analysis. Significant correlations of CTCs and DTCs were found with the clinicopathological parameters including the tumor stage (OR=1.95), depth of invasion (OR=1.99), lymph node metastasis (OR=2.44), distal metastasis (OR=5.98), histological differentiation (OR=1.67) and lymphovascular invasion (OR=4.48). CTCs and DTCs were also correlated with the prognostic events including relapse (RR=6.86) and metastasis (RR=3.22) and with the survival outcomes including the overall survival (OS) overall analysis (HR=3.46) and disease-free survival/progression-free survival (DFS/PFS) overall analysis (HR=3.00).

CONCLUSIONCTCs and DTCs are significantly associated with an advanced tumor stage, depth of tumor invasion, lymph node metastasis, distant metastasis before therapy, differentiation, lymphovascular invasion, relapse and metastasis in patients with EC. They are also significantly correlated with a poorer survival for OS and DFS/PFS to serve as clinical and prognostic predictors in patients with EC.

Disease-Free Survival ; Esophageal Neoplasms ; diagnosis ; Humans ; Lymphatic Metastasis ; Neoplasm Recurrence, Local ; Neoplastic Cells, Circulating ; Odds Ratio ; Prognosis ; Proportional Hazards Models ; Survival Analysis

OBJECTIVENucleostemin (NS) is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells, and maintains cell self-renewal. We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation.

METHODSNS mRNA and protein expression in A549, HepG2, SMMC-7721, HeLa, and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA (NC). The effect on cell proliferation was also analyzed by MTT assays.

RESULTSNS mRNA and protein were both expressed in A549 cells and four other tumor cell lines; the relative expression levels were similar in all five cell lines. The three pairs of NS siRNA, either transfected alone or cotransfected into A549 cells, could effectively inhibit the expression of NS mRNA and protein. Moreover, the interference ratio showed an obvious concentration-dependent relationship. NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%.

CONCLUSIONNS gene was not only highly expressed but also played an important role in A549 cell proliferation. Thus, targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.

Carcinoma ; genetics ; metabolism ; therapy ; Cell Line, Tumor ; Cell Proliferation ; Feasibility Studies ; GTP-Binding Proteins ; genetics ; Genetic Therapy ; Humans ; Lung Neoplasms ; genetics ; metabolism ; therapy ; Nuclear Proteins ; genetics ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering

The applications accepted and approved by general program, young scientist fund and fund for less developed region of national natural science funds in the discipline of Chinese materia medica, NSFC in 2012 have been introduced. The research contents of the funded projects in the popular research areas have been summarized and the problems in the applications have been analyzed to give a reference to the scientists in the field of Chinese materia medica.
China ; Financing, Organized ; organization & administration ; Humans ; Laboratory Personnel ; economics ; Materia Medica ; chemistry ; Medicine, Chinese Traditional ; economics ; Natural Science Disciplines ; economics ; manpower ; organization & administration

To investigate the prevalence of drug-resistance mutations, resistance to antiretroviral drugs, and the subsequent virological response to therapy in treatment-naive and antiretroviral-treated patients infected with HIV/AIDS in Henan, China, a total of 431 plasma samples were collected in Queshan county between 2003 and 2004, from patients undergoing the antiretroviral regimen Zidovudine + Didanosine + Nevirapine (Azt+Ddi+Nvp). Personal information was collected by face to face interview. Viral load and genotypic drug resistance were tested. Drug resistance mutation data were obtained by analyzing patient-derived sequences through the HIVdb Program (http://hivdb.stanford.edu). Overall, 38.5% of treatment-naive patients had undetectable plasma viral load (VL), the rate significantly increased to 61.9% in 0 to 6 months treatment patients (mean 3 months) (P＜0.005) but again significantly decrease to 38.6% in 6 to 12 months treatment patients (mean 9 months) (P＜0.001) and 40.0% in patients receiving more than 12 months treatment (mean 16 months) (P＜0.005). The prevalence of drug resistance in patients who had a detectable VL and available sequences were 7.0%, 48.6%, 70.8%, 72.3% in treatment-na(1)ve, 0 to 6 months treatment, 6 to 12 months treatment, and treatment for greater than 12 months patients, respectively. No mutation associated with resistance to Protease inhibitor (PI) was detected in this study. Nucleoside RT inhibitor (NRTI) mutations always emerged after non-nucleoside RT inhibitor (NNRTI) mutations, and were only found in patients treated for more than 6 months, with a frequency less than 5%, with the exception of mutation T215Y (12.8%, 6/47) which occurred in patients treated for more than 12 months. NNRTI mutations emerged quickly after therapy begun, and increased significantly in patients treated for more than 6 months (P＜0.005), and the most frequent mutations were K103N, V106A, Y181C, G190A. There had been optimal viral suppression in patients undergoing treatment for less than 6 months in Queshan,Henan. The drug resistance strains were highly prevalent in antiretroviral-treated patients, and increased with the continuation of therapy, with many patients encountering virological failure after 6 months therapy.

OBJECTIVETo explore the effect of ricin temperature response gel on breast cancer and its regulatory effect on immune function in rats.

METHODSRicin was purified by chromatography and identified by immunoblotting. The rat subcutaneously transplanted breast cancer model was established. Forty model rats with a tumor diameter of about 3.0 cm were subjected to the study. They were randomized into four groups equally: the model group and three treated groups (blank gel, ricin, ricin-gel) were administered with blank gel, ricin, and ricin temperature response gel via percutaneous intratumor injection, respectively. The tumor was isolated 10 days later for the estimation of tumor inhibition rate (TIR) by weighing, pathologic examination, and detection of tumor apoptosis-associated genes bcl-2 and bax with semiquantitative RT-PCR. Also, peripheral blood was obtained to test T-lymphocyte subsets, the killing function of lymphocytes, and the contents of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2). The outcomes were compared between groups.

RESULTSThe TIR in the ricin-gel group was 61.8%, with the pathologic examination showing extensive tumor tissue necrosis. Compared with the model group, after ricin temperature response gel treatment, bcl-2 expression was down-regulated, bax expression was up-regulated, CD4+ lymphocytes and CD4+/CD8+ ratio in peripheral blood were increased, the killing function of lymphocytes was enhanced, and the contents of TNF-α and IL-2 were elevated (P < 0.05 or P < 0.01).

CONCLUSIONIntratumor injection of ricin temperature-responsive gel showed significant antitumor effect on breast cancer and could enhance the immune function in the tumor-bearing rat.

Animals ; Antineoplastic Agents ; administration & dosage ; Apoptosis ; drug effects ; CD4-CD8 Ratio ; Disease Models, Animal ; Female ; Gels ; therapeutic use ; Immunohistochemistry ; Immunomodulation ; drug effects ; Injections, Intralesional ; Interleukin-2 ; immunology ; metabolism ; Mammary Neoplasms, Experimental ; drug therapy ; immunology ; pathology ; Random Allocation ; Rats ; Rats, Wistar ; Ricin ; administration & dosage ; Sensitivity and Specificity ; Temperature ; Tumor Necrosis Factor-alpha ; immunology ; metabolism

OBJECTIVETo investigate the effects of HCMV infection on phenotypes of parotid duct epithelial cells and relative mechanisms.

METHODSThe expressions of immediate early antigen of HCMV, pan cytokeratin and cathepsin D etc. were detected by immunohistochemical staining in tissues of parotid cytomegalic inclusion disease.

RESULTSCytokeratin which acts as an epithelial marker became negative while staining of Cathepsin D was intensified in parotid duct epithelial cells after infected by HCMV.

CONCLUSIONIt demonstrated that cytokeratin was lost through over-expression of Cathepsin D in parotid duct epithelial cells infected by HCMV.

Animals ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Antigens, Viral ; analysis ; Cathepsin D ; analysis ; Cytomegalovirus ; immunology ; physiology ; Cytomegalovirus Infections ; metabolism ; pathology ; virology ; Desmin ; analysis ; Epithelial Cells ; metabolism ; pathology ; virology ; Female ; Glial Fibrillary Acidic Protein ; analysis ; Host-Pathogen Interactions ; Humans ; Immunohistochemistry ; Infant ; Keratins ; analysis ; Male ; Mice ; Salivary Ducts ; metabolism ; pathology ; virology ; Vimentin ; analysis