Aim To isolate the female-specific sequence of Schistosomajaponicum S. J. in order to provide a basis for researching vaccines of anti-fecundity. Methods Representational difference analysis (RDA) was used to obtain a female-specific sequence of S.j. by the male S.j. genomic DNA substractive hybridization with the female. The fragment was about 562bp. Results This sequence was labeled as Dig probe by PCR technique to hybridize with the restriction fragments of each sex and the tester as well as the driver on the nylon membranes.The antiboly of Dig、 NBT、 and X-phosphate solution were utilized to immunoassay the result of hybridization. The result showed that the probe can hybridize to the tester but can not hybridize to the driver. Conclusion The result suggested that the fragment of approx. 562bp was the female-specific sequence of Schistosoma japonicum.

OBJECTIVE:To establish a method for the content determination of cabozantinib in its raw material. METHODS:RP-HPLC method was adopted. The determination was performed on Inertsil ODS-SP C18 column with mobile phase consisted of acetonitrile-0.02 mol/L ammonium acetate buffer (pH 5.2,52:48,V/V) at the flow rate of 1.0 mL/min. Detection wavelength was set at 241 nm,the column temperature was 38 ℃,and sample size was 20 μL. RESULTS:The linear range of cabozantinib were 9.88-49.40 μg/mL(r=0.9999). The limit of quantitation was 11.46 ng,and the limit of detection was 3.36 ng. The RSDs of preci-sion,stability,repeatability tests were all lower than 2.0%;recoveries were 98.5%-101.7%(RSD=1.2%,n=9). CONCLU-SIONS:The method is simple,accurate and suitable for the content determination of cabozantinib in its raw material.

Objective:To compare the drug release of nifedipine sustained-release tablets from four manufacturers to evaluate the intrinsic quality. Methods:The drug release rate was determined by UV respectively with pH 1. 2 HCl solution, pH 4. 0 sodium ace-tate buffer, pH 6. 8 phosphate buffer and water as the dissolution medium. The dissolution curves were compared by f2 factor. Results:The drug release rate of nifedipine sustained-release tablets from the four manufacturers all met quality standard of our country, al-though the dissolution curves in the different dissolution medium was various. Conclusion:There are differences in intrinsic quality a-mong the nifedipine sustained-release tablets from the four manufacturers. The dissolution examination standard should be improved fur-ther.

Objective:To study the feasibility of a new composite mineralized collagen membrane in alveolar ridge preservation.Methods:The third mandibular premolars on both sides were extracted from 12 dogs,the 24 alveolar sockets were randomly assigned into A,B and C groups(n =8).The distal alveolar sockets of group A was immediately implanted with the new composite mineralized collagen membrane and bone graft material,those of group B with bone graft material,group C was the blank control.The healing of sockets was evaluated by gross observation,morphological measurements,X-ray microscope and photographs of spiral CT.Results:The horizontal width of the alveolar process of group A was bigger than that of group B (P ＜ 0.05),that of group B was bigger than that of group C (P ＜ 0.05).The region of extraction interest in spiral CT value was higher in group A than that in group B and C (P ＜ 0.05).The extraction sockets were generated new bone and the degree of reconstruction measurements was higher in group A than that in group B and C (P ＜ 0.05).Conclusion:The new composite mineralized collagen membrane can induce the regeneration of new bone,and preserve the alveolar.

Objective To study the effect of tiotropium on urination disorder in benign prostatic hyperplasia (BPH) patients with chronic obstructive pulmonary disease (COPD).Methods In our prospective pilot study,96 BPH patients with COPD patients were enrolled as the treatment group and another 25 similar cases as the control group:In the former group tiotropium was administered and the control group was not.The two groups were compared in terms of the score by the international Prostate Symptom Score(IPSS),the quality of life by QOL,maximum flow rate (Q-max),average flow rate (Q-ave),time to Q-max (TTQ-Max),prostate volume (PVR) and bladder voiding efficiency (BVE) after six months treatment.Results As compared to the control,after six months treatment,such indexes in the treatment group as IPSS (15.1 ± 4.1,16.3 ± 3.4 and 14.7 ± 3.1,P =0.864),QOL(3.9 ± 0.8,4.0± 0.8 and 4.0 ± 0.9,P =0.992),Q-Max(ml/s) (8.5 ± 2.9,10.9 ± 2.2 and 9.0 ± 2.4,P =0.214),Q-ave(ml/s) (3.9 ±1.2,5.0 ± 1.4 and 3.8 ± 0.9,P =0.054),TTQ-Max(s) (11.1 ± 5.6,11.2 ± 4.0 and 10.4 ± 5.1,P =0.424),PVR(mL)(56.8 ± 33.3,62.3 ± 30.5 and 57.4 ± 29.5,P =0.981),BVE(％) (75.6 ± 13.8,72.7 ± 10.5 and 74.3 ± 12.1,P =0.992).showed no significant differences.Conclusion Tiotropium does not adversely affect lower urinary tract functions in BPH patients with COPD.

，and 17.47%.Conclusions Xylitol can lower the blood glucose a littte but without significant difference.It has little effect on blood glucose variability of patients with type 2 diabetes mellitus and can be safely used for rehydration.

ObjectiveTo evaluate the therapeutic efficacy of low-dosage methyltestosterone or andriol in men with senile osteoporosis. MethodsA total of 134 male patients with senile osteoporosis and the decreased serum level of free testosterone were divided into three groups. 45 patients were treated with low-dosage methyhestosterone(100 mg, once a day, sublingual) and 46 patients were treated with low-dosage andriol (40 mg, once a day, orally), while 43 patients were treated with placebo. The duration of treatment in each group was 1 year. The bone density, blood and urine biochemical indexes related to bone metaholites,the quality of life indexes, ultrasonography for prostate,serum prostate specific antigen,blood routine, urine routine, hepatic and renal function were detected before and after the treatment. ResultsBoth low-dosage methyltestosterone and low-dosage andriol could prevent the decrease of bone mineral density and improve patients' general health, role-emotional function and vitality (all P＜0.05). The difference values of femoral neck bone mineral density before and after treatment with low-dosage andriol and low-dosage methyltestosterone were (0.14+0.18)g/cm2 and (0.12±0.09)g/cm2 , respectively(P＜0.05). Low-dosage andriol hadstronger effects in increasing the level of estradiol (32.5±14.2 )ng/L than low-dosage methyltestosterone(19.3±9.2)ng/L(P＜0.05) and showed more notable effects in improving the physical functioning and role-physical function than low-dosage methyhestosterone. The use of the two androgenic hormones at low dosage showed safety. ConclusionsBoth low-dosage methyltestosterone and low-dosage andriol can be used to treat senile osteoporosis in men and to improve life quality. Both of them are effective and safe therapeutic choices.

Objective To detect the protective mechanism of trehalose in tracheal cryopreservation.Methods Inbred male Sprague-Dawley (SD) rats were sacrificed with intraperitoneal injection of ketamine(150mg?kg -1).The tracheas were removed and immersed immediately in the freezing medium of low potassium dextran (LPD) solution only(Group Ⅰ) ,containing with 10% dimethylsulfoxide(DMSO)(Group Ⅱ), containing with 0.15mol?L -1 trehalose (Group Ⅲ),and containing with 10% DMSO and 0.15mol?L -1 trehalose (Group Ⅳ) respectively. A sterile plastic tube containing a 1-cm-long trachea was filled with the freezing medium,sealed,and frozen to -80℃ at rate of -1℃ per minute in a programmable freezer.Then the tube was stored in liquid nitrogen(-196℃) for 20 days. Then the specimen was thawed in a 37℃ water bath and rinsed with physiologic saline solution 10 times.Histologic changes before cryopreservation and after thawing were examined in each group. After the specimens were embedded in paraffin,5-(m-thick sections were stained with hematoxylin and eosin.The epithelium and cartilage was assessed. We also observed Bcl-2 and Bax gene expression by immunohistochemistry. At last, some tracheas(SD) after cryopreservation were thawed and transplanted into the abdominal cavity of Wistar rats. The transplanted tracheas were retrieved and assessed histologically.Results Microscopic findings of the tracheas in Group Ⅲ and Group Ⅳ showed their structure were intact and Bax gene expression was lower in cartilage after cryopreservation(20d) compared with other groups,especially in Group Ⅳ.The tracheas in Group Ⅲ and Group Ⅳ grew well after they were transplanted into cavity of Wistar rats heterotopically,too.There were no significant differences among 4 groups in Bcl-2 gene expression.Conclusion In tracheal cryopreservation the trehalose can protect the trachea by protecting the tracheal cartilage.It is one of the protective mechanism that the trehalose inhibit the Bax gene expression of cartilage cells.The concomitant use of trehalose and DMSO has a synergistic effect.

AIM:To explore the molecular mechanism through which curcumin reverses hepatocyte growth factor (HGF)-induced resistance to gefitinib in lung cancer cells.METHODS:The methods of MTT assay, wound healing assay and Western blot were used to observe the effects of HGF, curcumin and gefitinib on the migration, drug susceptibility, epithelial-mesenchymal transition, and related signaling pathways in the PC9 lung cancer cells.RESULTS:HGF reduced susceptibility of the PC9 cells to gefitinib, and curcumin significantly reversed HGF-induced resistance to gefitinib.HGF induced migration and epihelial-mesenchymal transition, and promoted c-Met/AKT/mTOR pathway activation in the PC9 cells.Gefitinib alone did not prevent the above activities.However, combined with curcumin, gefitinib prevented the above activities.CONCLUSION:Curcumin reverses HGF-induced resistance of the PC9 cells to gefitinib by preventing epithelial-mesenchymal transition and inhibiting c-Met/AKT/mTOR activation.

For rapid and accurate screening of recombinant modified vaccinia virus Ankara (rMVA) that satisfied the quality standards of clinical trials, a novel shuttle vector that can delete the marker gene automatically during virus propagation was construted: pZL-EGFP. To construct the pZL-EGFP, the original shuttle vector pSC11 was modified by replacing the LacZ marker gene with enhanced green fluorescent protein (EGFP) and then inserting homologous sequences of TKL into the flank regions of EGFP. Baby hamster kidney (BHK)-21 cells were cotransfected with pZL-EGFP and MVA, and underwent ten passages and one plaque screening to obtain the EGFP-free rMVA carrying the exogenous gene. Resulting rMVA was tested by polymerase chain reaction and western blotting to verify pZL-EGFP function. A novel shuttle vector pZL-EGFP containing an EGFP marker gene which could be deleted automatically was constructed. This gene deletion had no effect on the activities of rMVA, and the exogenous gene could be expressed stably. These results suggest that rMVA can be packaged efficiently by homologous recombination between pZL-EGFP and MVA in BHK-21 cells, and that the carried EGFP gene can be removed automatically by intramolecular homologous recombination during virus passage. Meanwhile, the gene deletion had no influence on the activities of rMVA and the expression of exogenous target gene. This study lays a solid foundation for the future research.
Animals ; Biomarkers ; Cricetinae ; Epithelial Cells ; virology ; Gene Deletion ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombination, Genetic ; Vaccinia ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication