Humans ; Lead ; toxicity ; Male ; Reproduction ; drug effects ; Sperm Count ; Sperm Motility ; drug effects

The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.

Objective To investigate the susceptibility of amphotericin B,itroconazol and voriconazole against filamentous fungi.Methods Etest was used to determine the MIC of amphotericin B, itroconazol and voriconazole against filamentous fungi including Aspergillus,Penicillium,Alternaria alternate,Mucor and Rhizopus species.Results The average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus fumiagtus is O.29 ?g/ml,1.16 ?g/ml and 5.88 ?g/ml;the average MIC of amphotericin B and voriconazol to Aspergillus flavus is 6.39 ?g/ml and 0.22 ?g/ml;the average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus niger is 0.69,2.31,and 19.75 ?g/ ml.Most of Penicillium are susceptable to amphotericin B,but 3 strains showed very high MIC to voriconazol and itroconazol.Both of the testing strains of Mucor and Rhizopus were resistant to all of the three antifungal agents.Conclusion Amphotericin B,itroconazol and voriconazole possessed different susceptibility on different types of filamentous fungi.It is important for clinical laboratories to identify the filamentous fungi to the level of genus and species to help physicians choose antifungal agents correctly.

Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.

Adult ; Aged ; Coal Mining ; Health Promotion ; methods ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Young Adult

OBJECTIVETo observe the sodium current of acutely isolated alveolar type II cells and its regulation.

METHODSAfter isolation the AT II cells, the whole cell sodium current were recorded in 5 hours with patch-clamp in whole-cell mode, and the effect of amiloride and terbutaline on the current was investigated.

RESULTSThe amiloride-sensitive current of AT II cells were recorded, which could be obviously stimulated by terbutaline.

CONCLUSIONAcutely isolated AT II cells can be valuable for studying electrophysiological characteristics of sodium channels even under pathological conditions.

Amiloride ; pharmacology ; Animals ; Biological Transport ; Cell Separation ; Patch-Clamp Techniques ; Pulmonary Alveoli ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism ; Sodium Channel Blockers ; pharmacology ; Sodium Channels ; metabolism ; physiology ; Terbutaline ; pharmacology

Establishment of quality management system (QMS) plays a critical role in the clinical data management (CDM). The objectives of CDM are to ensure the quality and integrity of the trial data. Thus, every stage or element that may impact the quality outcomes of clinical studies should be in the controlled manner, which is referred to the full life cycle of CDM associated with the data collection, handling and statistical analysis of trial data. Based on the QMS, this paper provides consensus on how to develop a compliant clinical data management plan (CDMP). According to the essential requirements of the CDM, the CDMP should encompass each process of data collection, data capture and cleaning, medical coding, data verification and reconciliation, database monitoring and management, external data transmission and integration, data documentation and data quality assurance and so on. Creating and following up data management plan in each designed data management steps, dynamically record systems used, actions taken, parties involved will build and confirm regulated data management processes, standard operational procedures and effective quality metrics in all data management activities. CDMP is one of most important data management documents that is the solid foundation for clinical data quality.
Clinical Trials as Topic ; Data Collection ; standards ; Database Management Systems ; standards ; Information Storage and Retrieval ; standards

OBJECTIVETo observe the changes of sodium transport of alveolar type II cells in oleic acid-induced acute respiratory distress syndrome (ARDS).

METHODSAfter isolation of the AT II cells as ARDS model induced by oleic acid, the whole cell sodium current was recorded in 5 h with patch-clamp in whole-cell mode. The effect of beta2-agonist terbutaline on the current was also examined.

RESULTSThe sodium currents recorded from AT II cells in ARDS were smaller than that of the control cells, but the current could be stimulated to increase obviously by terbutaline.

CONCLUSIONAT II cells can actively transport sodium ion even under conditions of ARDS, but this capacity can not match that of the control cells. Terbutaline can obviously stimulate the sodium current.

Acute Disease ; Animals ; Ion Transport ; Lung Diseases ; chemically induced ; metabolism ; physiopathology ; Male ; Membrane Potentials ; Oleic Acid ; Patch-Clamp Techniques ; Pulmonary Alveoli ; metabolism ; pathology ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism ; Sodium Channels ; physiology ; Syndrome

OBJECTIVETo investigate the effect of S-phase kinase-associated protein 2 antisense oligodeoxynucleotide (Skp2 ASODN) on the growth and proliferation of gastric carcinoma SGC-7901 cells and its mechanism.

METHODSThe Skp2 oligodeoxynucleotides (ODNs) were embedded in cationic liposome Lipofectamine 2000 reagent and transfected into SGC-7901 cells. The cell growth and proliferation were observed with light microscopy and MTT assay. Cell cycle was measured by flow cytometry. The expression levels of Skp2 and p27 mRNA were detected by reverse transcription-polymerase chain reaction. The expression levels of Skp2 protein and its substrate p27 protein were detected by Western blot.

RESULTAfter treatment with Skp2 ASODN, the growth and proliferation of SGC-7901 cells were inhibited in a dose-dependent manner with a peak value at 48 h. The inhibition rate of 200 nmol/L group at 48 h was 42.4 % (P<0.01). In cell cycle study the percentage of S phase cells in 200 nmol/L group was significantly higher than that in normal control group (P<0.05). Both Skp2 mRNA and its protein levels in 200 nmol/L group were significantly lower than those in control group and in Skp2 nonsense oligodeoxynucleotide (Skp2 NSODN) group (P<0.05). However, p27 mRNA level remained unchanged although its protein level was significantly higher than that in control group and NSODN group (P<0.05).

CONCLUSIONSkp2 ASODN can inhibit the growth and proliferation of SGC-7901 cells, which may be mediated by interfering with ubiquitin-proteosome pathway and cell cycle regulation.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Oligodeoxyribonucleotides, Antisense ; genetics ; pharmacology ; S-Phase Kinase-Associated Proteins ; genetics ; pharmacology ; Stomach Neoplasms ; pathology ; Transfection