Yifeng (TE 17), Tinghui (GB 2), Zhongzhu (TE 3) and Xiaxi (GB 43) were selected as key acupoints plus hyperbaric oxygen to treat 50 cases of sudden deafness, and the total effective rate was 98%.

Objective To study the inhibition effect of leech extract on HepG2 cells. Methods Human hepatocellular cancer cell line HepG2 were treated with different concentrations of leech extracts which were extracted by method of freeze-thawing with liquid nitrogen and contrasted with that by method of water extracting and ethanol precipitating. The inhibition effects and cell morphous were examined by MTT assay and Acridine orange (AO) fluorescent staining method respectively. Result The 6～15 mg/mL drug concentrations of leech extract by method of freeze-thawing with liquid nitrogen had an obvious inhibition on proliferation of HepG2 cells in a dose-dependent manner, and the effect was better than that by method of water extracting and ethanol precipitating (P

In the present study, the expression, purification, toxicity detection and the adjuvant effect of the heat-labile enterotoxin in non-toxic mutant mLT63 of Escherichia coli were investigated, in which the inductive expression was performed under optimal condition for inductive expression and the toxicity of the products obtained from inductive expression were tested for toxicity after being purified and concentrated with affinity chromatography. BALB/c mice were immunized orally with the mutant mLT63 associated with Helicobacter pylori (Hp) subunit vaccine UreB, Omp11. After immunization, the specific antibody levels in serum, extract from gastric tissues and fecal extract were determind by means of ELISA assay and the results were subjected to statistical analysis. It was demonstrated that the mutant mLT63 of heat-labile entrotoxin of E.coli constructed in our laboratory devoided of any toxic effect as revealed by the rabbit ileal loop assay, but its adjuvant effect could be demonstrated in the associated immunization of mice with Hp subunit vaccine UreB and Omp11.

Objective To design an instrument that can provide a series of pulse to stimulate cell by means of external electric field, the structure and function of organism can retrieve. Methods AVR MCU, produced by America ATMEL Co., is used as the core of the system. The program has been developed to adjust three pulse's parameters, including amplitude, frequency and pulse duration. The cooperation between DAC and OP completes the transformation from monopole pulse to bipolar pulse. The booster PB50 amplifies the current and voltage of the output. Results The Stimulator can provide bipolar pulse, amplitude: up to ?40V, frequency: 0.01Hz -10Hz, pulse duration: 0.4ms -24ms. Conclusion Cooperating with special electrode board, the instrument can provide effective electric field for cell simulating. At present the instrument has been used in the research of the endothelial progenitor cell.

Objective To investigate the effects of anaesthetic concentration of sevoflurane on TM3 mouse leydig cell viability.Methods TM3 mouse leydig cells were randomly divided into 3 groups ( n =24 dishes each):control group (group C),2％ and 5％ sevoflurane groups (groups SEV1 and SEV2 ).The cells were collected after being exposed to sevoflurane or 95 ％ room air + 5 ％ CO2 for 2,4 or 6 h (T1-3) for microscopic examination with optical binocular inverted microscope.The number of live cells was counted by using cell counting kit8.Gene chips were used to indentify differentially expressed genes between group C and group SEV2 after being exposed to air and 5 ％ sevoflurane for 6 h respectively.Results The leydig cell viability was significantly decreased at T3 in group SEV2 as compared with groups C and SEV1.Morphological changes were found only in group SEV2.A total of 45 genes were identified to be differentially expressed in group SEV2 as compared with group C.The level of expression of prostaglandin-endoperoxidase synthase 2 gene (Ptgs2),chemokine (C-C motif) ligand 2(CCL2) gene and dual specificity phosphatase1 (Dusp1) gene increased by at least 4 times in group SEV2.Conclusion Sevoflurane can inhibit the cell viability of TM3 mouse leydig cell in concentration dependent manner through abnormal expression of Ptgs2,CCL2 and Dusp1 genes.

Objective To investigate the effects of inhalation anesthetics on human sperm motility and capacitation in vitro. Methods Sperm samples were obtained from normal adults and prepared with discontinuous percoll gradient centrifugation technique. The samples were incubated for 5 h in an airtight glass container filledwith 5% CO2-95% air at 37 ℃ with or without sevoflurane (SEV 2%, 4% ) or isoflurane (ISO 1.1%, 2.2% ).Then human sperm motility was examined in vitro at 37℃ and analyzed by the computer-assisted sperm analysis (CASA), including sperm motility (a + b)%, curvilinear velocity (VCL), straight line velocity (VSL), averagepath velocity (VAP) and amplitude of lateral head displacement (ALH). The capacitation effect was assessed by using the chlortetracycline (CTC) staining and phase-contract microscopy. Results 2% and 4% SEV significantly reduced (a + b)% , VCL, VSL and VAP in a dose-dependent manner, while only 4% SEV significantly decreased ALH and the capacitation ability of the sperm compared with control group. 2.2% ISO significantly decreased ( a + b)%, VCL, VSL and VAP compared with control and 1.1% ISO group. The capacitation ability of the sperm was significantly decreased by 1.1% and 2.2% ISO as compared with control group. Conclusion Sevoflurane and isoflurane have significant inhibitory effects on human sperm motility and capacitation in a dose-dependent manner. Sevoflurane has stronger inhibitory effect than isoflurane.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To investigate the immunization effect of influenza A/H1N1 vaccine in health care workers (HCW) in Inner Mongolia Greater Khingan Mountains area. Methods Five hundred and five HCW who received A/H1N1 influenza vaccination (immunized group) and 129 staffs who didn't receive the vaccination (unimmunized group) were randomly sampled for semiquantitative testing of serum H1N1 antibody (IgG) levels by enzyme-linked immunosorbent assay (ELISA).Results were analyzed and stratified by age, sex, occupation and the time interval between the time of vaccination and serum sample collection. The antibody positive rates of the two groups were compared by x2test. Results There were 401 (79. 4%) HCW whose H1N1 antibody were positive and 50 (9.9%) whose antibody were weak positive among 505 immunized HCW. While among 129 unimmunized HCW, there were 59 (45.7%) whose antibody were positive and 15 (11.6%) whose antibody were weak positive. The seroconversion rates of specific antibody were not significantly different among the different age groups after receiving A/H1N1 influenza vaccine (P＞ 0.05).However, there were statistical differences of the seroconversion rates among different sex groups (men 95.7% vs women 87.4% in immunized group, x2=6.40, P＜0.05; and men 73.3% vs women 52.5% in unimmunized group, x2 =4.07, P＜0.05) and different occupation groups (doctor 86.0% vs nurse 94.5% in immunized group, x2 = 9. 16, P＜0.01; and doctor 43. 8% vs nurse 75.0% in unimmunized group, x2=12.61, P＜0.01 ). The seroconversion rate was 81.5% after 80 to 89 days of vaccination, which was significantly lower than those after 30 to 39, 50 to 59 days and 60 to 69 days of vaccination, which was 100.0%, 94.7% and 93.6%, respectively (x2 =3.96, P ＜0.05; x2=7.15, P ＜0. 01; x2 = 9. 98, P＜0. 01). Conclusions A/H1N1 influenza vaccination can induce effective immune response in HCW in Greater Khingan Mountains area of Inner Mongolia. However,the level of specific antibody significantly reduces after 80 to 89 days of vaccination.

Group-A rotaviruses are recognized as the most common cause of acute diarrhea. Phylogenetic analyses of the VP7 genes of rotaviruses circulating in Nanjing (China) could aid in the development of rotavirus vaccines. A total of 908 stool specimens were collected from patients suffering from acute diarrhea in Nanjing between October 2012 and December 2013, and were tested further for rotaviruses. Fifty rotavirus isolates selected randomly were typed by reverse transcription-polymerase chain reaction using serotype-specific primers for G genotyping. VP7 genes of 19 G9 strains were sequenced for further genetic characterization. Among the 908 stool specimens examined during the surveillance period, 103 (11.34%) were rotavirus-positive. G9 was the most predominant genotype (78.0%), followed by G2, G1 and G3. Sequence and phylogenetic analyses of the VP7 genes of serotype G9 rotaviruses revealed these strains to comprise two lineages (G9-VI, G9-III) and to be dominated by the G9-VI lineage (which belonged to a unique subcluster of Japanese and Chinese G9 strains). Amino-acid sequences of the four antigenic regions (A, B, C or F) were variant among a portion of strains, which may have contributed to the prevalence of G9 rotaviruses in this area.
Adult ; Amino Acid Sequence ; Antigens, Viral ; chemistry ; genetics ; Capsid Proteins ; chemistry ; genetics ; China ; Evolution, Molecular ; Humans ; Infant ; Molecular Sequence Data ; Mutation ; Phylogeny ; Rotavirus ; genetics ; immunology ; physiology ; Serogroup

Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.