OBJECTIVE:To determine the content of ?-schizandrin,one of the effective ingredients,in Shuangjia Wuling capsules METHODS:The RP-HPLC method was performed with YWG C18 column(4 6mm?250mm) The mobile phase consisted of methanol-water(72∶28) The detecting wavelength was 254nm RESULTS:The calibration curve for ?-schizandrin was linear in the range of 0 0 207～0 4 130mg/ml(r=0 9 999) The average recovery was 97 68% with RSD=1 85% CONCLUSION:The method is simple,rapid and reliable for quality control of the capsules

AIM: To investigate the effect of testosterone with varied concentrations on the fibr inolysis activity of HUVEC and its mechanism. METHODS: Human umbilical vein endothelial cells (HUVEC) were cul tured as recommended. After confluence, the cultures were treated with testoster one(3 ?10 -10, 3?10 -9, 3?10 -8,3?10 -6, 3?10 -5 m ol/L) , and the control confluent cells were cultured in the same medium witho ut steroid. MTT experiment was repeated for 72 hours to investigate each groups' cell proliferation. The tPA and PAI-1 antigen levels were assayed with ELISA K its. Then with HUVEC incubated in androgen receptor antagonist (flutamide) 3 hou rs previously, the experiment was repeated. RESULTS: Testosterone at physiologic or lower concentrations (3 ?10 -10 to 3?10 -8 mol/L ) stimulated the secretion of tPA by HUVEC (P

Objective To establish a method of reverse hybridization to detect five subfamilies of low risk Human Papillomaviruses(HPV6,11,42,43 and 44)and eighteen subfamilies of high risk HPV (HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83 and MM4)in one reaction.Methods Special probes for twenty-three HPV subfamilies were fixed on nylon membrane bars,biotin labeled general primers mediated polymerase chain reaction(GP-PCR)were applied in HPV DNA amplification.PCR amplified DNA fragments were reversely hybridized with special probes that were fixed on the membranes. All samples(136)detected by reverse hybridization method were paralleled with the methods of Hybridization Capture Ⅱ(HC-Ⅱ)and sequencing.Results Positive rate of the 136 samples detected by reverse hybridization was 41.9%,while HC-Ⅱ 42.6% and sequencing 40.4%.Reverse hybridization detection indicated coherence with the other two methods(Kappa 0.8644 and 0.9089,respectively).While sequencing was lab standard for DNA test,the sensitivity was 96.36%,specificity was 95.06%,accuracy was 95.59%.Conclusions Method of reverse hybridization is adaptable to 23 kinds of HPV subfamilies, which can confirm the exactly subfamilies of HPV infection.This method is adaptable in clinical detection of HPV,with high sensitivity,high specificity,simply and convenient operation and the results are easily to be read.

DNA repair processes play a role in the development of drug resistance which represents a huge obstacle to leukemia chemotherapy.Histone H2AX phosphorylation (ser139) (γH2AX) occurs rapidly at the onset of DNA double strand break (DSB) and is critical to the regulation of DSB repair.If DNA repair is successful,cells exposed to anti-neoplastic drugs will keep entering the cycle and develop resistance to the drugs.In this study,we investigated whether γH2AX can be used as an indicator of tumor chemosensitivity and a potential target for enhancing chemotherapy.K562 and multi-drug resistant cell line K562/A02 were exposed to adriamycin (ADR) and γH2AX formed.Flow cytometry revealed that percentage of cells expressing γH2AX was increased in a dose-dependent manner and the percentage of K562/A02 cells was lower than that of K562 cells when treated with the same concentration of ADR.In order to test the potential of γH2AX to reverse drug resistance,K562/A02 cells were treated with PI3K inhibitor LY294002.It was found that LY249002 decreased ADR-induced γH2AX expression and increased the sensitivity of K562/A02 cells to ADR.Additionally,the single-cell gel electrophoresis assay and the Western blotting showed that LY249002 enhanced DSBs and decreased the expression of repair factor BRCAl.These results illustrate chemosensitivity can partly be measured by detecting γH2AX and drug resistance can be reversed by inhibiting γH2AX.

DNA repair processes play a role in the development of drug resistance which represents a huge obstacle to leukemia chemotherapy. Histone H2AX phosphorylation (ser139) (γH2AX) occurs rapidly at the onset of DNA double strand break (DSB) and is critical to the regulation of DSB repair. If DNA repair is successful, cells exposed to anti-neoplastic drugs will keep entering the cycle and develop resistance to the drugs. In this study, we investigated whether γH2AX can be used as an indicator of tumor chemosensitivity and a potential target for enhancing chemotherapy. K562 and multi-drug resistant cell line K562/A02 were exposed to adriamycin (ADR) and γH2AX formed. Flow cytometry revealed that percentage of cells expressing γH2AX was increased in a dose-dependent manner and the percentage of K562/A02 cells was lower than that of K562 cells when treated with the same concentration of ADR. In order to test the potential of γH2AX to reverse drug resistance, K562/A02 cells were treated with PI3K inhibitor LY294002. It was found that LY249002 decreased ADR-induced γH2AX expression and increased the sensitivity of K562/A02 cells to ADR. Additionally, the single-cell gel electrophoresis assay and the Western blotting showed that LY249002 enhanced DSBs and decreased the expression of repair factor BRCA1. These results illustrate chemosensitivity can partly be measured by detecting γH2AX and drug resistance can be reversed by inhibiting γH2AX.

Objective Helicobacter pylori（ ）， To investigate the infection status of HP and analyze the correlation between HP Methods infection and serum bilirubin in railway drivers. A total of 2 731 railway drivers in Zhengzhou locomotive depot were - selected as study subjects using judgment sampling method. Carbon 13 urea breath test was used to evaluate the HP infection ， status. The metabolic indexes of HP positive group and HP negative group were compared and the relationship between HP Results （ ） ， infection and serum bilirubin was analyzed. The HP infection rate was 42.3% 1 156/2 731 . The older the age the ， （ ）， （ P ） longer the work years and the higher the body mass index BMI the higher the HP infection rate all <0.01 . The infection （P ） rate of HP in married people was higher than that in unmarried people <0.01 . The HP infection rate of smokers was higher - （P ） - ， than that of non smokers <0.01 . Compared with the HP negative group fasting blood glucose and serum levels of total ， （ - ）， （ ） - cholesterol low density lipoprotein cholesterol LDL C triglyceride and homocysteine Hcy were increased in the HP （ P ） （ - ）， ， positive group all <0.05 . The serum levels of high density lipoprotein cholesterol HDL C total bilirubin direct bilirubin （ ） - （ P ） DBIL and indirect bilirubin were lower than those in HP negative group all <0.05 . Logistic regression analysis showed that （ P ） HP infection was associated with low serum total bilirubin and low DBIL all <0.01 after adjusting for the confounding effects ， ， ， ， ， ， ， - ， - ， of age work years marital status smoking history fasting blood glucose total cholesterol triacylglycerol LDL C HDL C Conclusion ， ， ， and Hcy. The age work length BMI smoking and marital status are the influencing factors of HP infection in railway drivers. HP infection is associated with low levels of total bilirubin and DBIL.

AIM　To explore whether dexamethasone-dextran (260 000) has the characteristics of site-specific delivery in rat gastrointestinal tract. METHODS　Dexamethasone prodrug and dexamethasone were administered to rat ig at the dose of 5 μmol*kg-1. The distribution of dexamethasone in the contents and mucosa of different parts of the rat GI tract at different time intervals and its concentration in plasma were determined by HPLC. RESULTS　Dexamethasone was mainly released in the cecum and colon contents and mucosa after oral administration of dexamethasone prodrug. The absorption was reduced significantly. The peak time of the drug in plasma was 8.1 h, and the peak concentration was 32 μg*L-1. However, free dexamethasone was found mainly in the contents and mucosa of the stomach, proximal and distal small intestine. The peak time of the drug in plasma was 2.2 h, and the peak concentration was 2120 μg*L-1. CONCLUSION Dexamethasone can be specifically delivered to the large intestine by using dexamethasone-dextran (260 000). It appears that the prodrug has a potential in the treatment of inflammatory bowel disease.

To construct a secretory-expression vector of antimicrobial peptide Bactenecin 7(Bac7),and identify the secretory-expression product in L.lactis MG1363 and its bioactivity.The splicing primers of regulation elements and Bac7 gene,which designed according to codon usage preferences of L.lactis MG1363,were chemically synthesized,and the overlap-extension PCR method was used to splice the full length of Bac7 gene.Then the Bac7 gene was linked to expression vector pMG36e to construct pMG36e/Bac7 vector,and pMG36e/Bac7 was transformed into L.lactis MG1363 by electrophoration.RT-PCR and Western blot assays were applied to investigate the expression of the Bac7 gene in L.lactis,and bioactivity of Bac7 in culture supernatant of L.lactis was tested with plate-diffusion method.The results showed that the Bac7 gene and its regulation elements was amplified and cloned in the vector pMG36e successfully,The secretory-expressed Bac7 in L.lactis MG1363 harboring pMG36e/Bac7 was identified by Western blot,and it had high bacteriostatic activity against E.coli.These results indicate that the recombinant L.lactis MG1363 could express bioactive Bac7,which lays a foundation for further study of oral administration of a Bac7-secreting L.lactis to treat intestinal bacteria infection.

Objective To investigate the effect of three Scutellaria baicalensis extracts (baicalin,baicalein and wogonin) on the apoptosis of a human cutaneous squamous cell carcinoma cell line,SCL-1.Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to detect the proliferation of cultured SCL-1 cells treated with different concentrations (12.5,25,50,75 and 100 μmol/L) of baicalin,baicalein and wogonin for various durations (12,24 and 48 hours).Some SCL-1 cells were treated with baicalin,baicalein and wogonin of 12.5 μ mol/L respectively for 48 hours followed by the detection of cell apoptosis by double staining with annexin V-fluorescein isothiocyanate/propidium iodide in combination with enzyme-linked immunosorbent assay (ELISA),as well as estimation of cell cycle by flow cytometry.The 50％ inhibitory concentration was determined by line regression model and inner insert method,and statistical analysis was carried out by Student's t test,one-way analysis of variance (ANOVA),and Student-Newman-Keuls-q test.Results Baicalin,baicalein and wogonin all inhibited the proliferation of SCL-1 cells in a dose-and time-dependent manner (all P ＜ 0.01).Under the same conditions (treatment concentration and duration),baicalein showed the strongest inhibitory effect on the proliferation of SCL-1 cells,followed by wogonin and baicalin (all P ＜ 0.01).All the Scutellaria baicalensis extracts induced the apoptosis of SCL-1 cells and arrested them in G1-phase.The percentage of cells in G1 phase was 56.37％ ± 2.41％,74.23％ ± 2.02％ and 64.15％ ± 1.87％,and early apoptosis rate was 8.09％ ± 1.02％,24.13％ ± 0.76％ and 14.45％ ± 1.57％,in SCL-1 cells treated with baicalin,baicalein and wogonin of 12.5 μmol/L for 48 hours,respectively,compared to 45.04％ ± 1.93％ and 4.12％ ± 0.29％ in the untreated control cells respectively (F =83.29,186.37,respectively,both P ＜ 0.01).Similarly,there was a downward trend from baicalein to wogonin and baicalin in the effect on cell apoptosis and cell cycle arrest of SCL-1 cells.Conclusions Scutellaria baicalensis can inhibit the growth and induce the apoptosis of SCL-1 cells,which may provide new ideas for the treatment of cutaneous squamous cell carcinoma with traditional Chinese drugs.

Objective To explore weaning dependent patients off mechanical ventilation through rehabilitative treatment. Methods Eighty-four patients dependent on mechanical ventilation were randomly divided into a rehabilitation group (n =44) and a control group (n =40). All were treated with routine medication, but in addition patients in the rehabilitation program followed a rehabilitation program. Three weeks later, the success rates and the time of weaning off mechanical ventilation were observed in the two groups. Results In the rehabilitation group,88.6% of the patients were successfully weaned off mechanical ventilation, versus 60% of the control group. The respective average weaning times were 6.8 ± 3.8 days and 14.3 ± 5.2 days. Both differences were statistically significant. Conclusions Rehabilitative treatment improves the weaning success rate and reduces the time needed for weaning mechanical ventilation-dependent patients.