Background and purpose:Aberrant expressions of microRNAs (miRNA, miR)are reported in various cancers and may associate with cancer occurrence, development, invasion and metastasis, thereby functioning as either tumor suppressors or oncogenes. This study attempted to observe the expression of miR-214 in pancreatic cancer and to explore its clinical signiifcance.Methods:Real-time PCR was used to detect the miR-214 expressions between pancreatic cancer tissues and matched adjacent tissues. The correlations of miR-214 expression with clinic-pathological features and clinical prognosis were analyzed.Results:MiR-214 expression was up-regulated in 69.4%(25/36) of tumor tissue specimens. The relative expression level of miR-214 was signiifcantly higher in tumor tissues than in matched adjacent tissues (3.45vs 1.52,P<0.01). Higher miR-214 level was strongly associated with T3-T4 stage (P=0.018). The Kaplan-Meier analysis revealed that patients with higher expression of miR-214 had a shorter survival time (P=0.032).Conclusion:The expression of miR-214 is associated with clinic-pathological features and patient’s clinical prognosis, so it may be used as a potential diagnostic biomarker and a prognostic predictor in patients with pancreatic cancer.

Objective:To establish the quality standard for two effective components in extractum glycyrrhizae capsules. Methods:Radix glycyrrhizae was identified by a TLC method. The contents of liquiritin and ammonium glycyrrhizinate in extractum glycyrrhizae capsules were determined by HPLC. An Inertsil C18 (150 mm × 4. 6 mm, 5μm) column was used. The mobile phase consisted of ace-tonitrile (A)-0. 2％ phosphoric acid (B) (0-8 min: 20％A-20％A;8-34 min: 20％A-50％A;34-35 min: 50％A-100％A;35-40 min:100％A-20％A) at a flow rate of 1. 0 ml·min-1 . The detection wavelength was at 237 nm under 25℃. Results: The spots in TLC were clear. Liquiritin showed a good linear relationship within the range of 0. 0020-0. 1000 mg·ml-1(r=0. 9995). The aver-age recovery was 100. 29％, and the RSD was 2. 94％(n=6). Ammonium glycyrrhizinate showed a good linear relationship within the range of 0. 0020-0. 1000 mg·ml-1(r=0. 9998). The average recovery was 101. 46％, and the RSD was 2. 33％(n=6). Conclu-sion:The method is simple,reliable and reproducible, which can be used for the quality control of the preparation.

Objective:To establish a trace chromogenic substrate method for the determination of anti-FIIαactivity in compound heparin sodium cream .Methods: The anti-FIIαactivity in compound heparin sodium cream was determined by a trace chromogenic substrate method according to the completely random design of experiment based on the amount reaction principle of 4*4 parallel lines in the biological test statistics method .Results:The calibration curve was linear within the range of 0.005 04 IU· ml-1-0.021 IU· ml-1(r=0.992).The average recovery was 101.6% with RSD of 2.76% (n=9).Conclusion: The method is accurate, reliable and reproducible , and can be used for evaluating the quality of compound heparin sodium cream .

Objective:To investigate the effects of endostatin on Ang-2/Tie-2 system in ectopic lesion.Methods:Thirty mice of endometriosis model (EM) were established by peritoneum planted.After 1 week of transplantation,model mice were randomly divided into three groups including rAAV2-hEndostatin-EGFP group (n=20),rAAV2-EGFP group (n=20)and PBS control group (n=20).The histology changes were examined in the three groups.The expressions of Ang-2 and Tie-2 Were examined by immunohistochemistry.Results:The EM model mice were established suecessfully.The gland proliferation was obvious in the two control groups.The mesenchymal blood vessel was rich.But there were gland atrophy and blood vessel reducing in endostatin group.There were expressions of Ang-2 and Tie-2 in the cytoplasm of endothelium,epithelial glands and stroma.The positive rate of Ang-2 expression was 40%in endostatin group,much lower than that in other two groups(75%and 80%,P＜0.05).The positive rate of Tie-2 expression was 45%in endostatin group,which was also lower than that in other two groups(80%and 85%,P＜O.05).Conclusion:Endostatin can effectively interfere angiogenesis process of Ang-2/Tie-2,which lays the foundation for clinical therapy by anti-angiogenesis.

Objective:To investigate the effect of human interferon-α-2b(IFN-α-2b) in the treatment of endometriosis by detecting the levels of CA125 and endometrial antibody(EMAb). Methods: Forty-five cases with endometriosis were divided into three groups. Fifteen cases in test group (operation+IFN-α-2b), 20 cases in control group A (operation+Diphereline), 10 cases in control group B(only operation). The blood serum level of CA125 was detected by microparticle enzyme immunochemiluminescent at pre-operation, the first month, second month and third month of the post-operation; and the EMAb level was measured by enzyme linked immunosorbent assay (ELISA). The follow-up of patients was carried out after operation to evaluate the clinical appearances and record the adverse effects of the human IFN-α-2b. Results: Compared with pre-operation, the levels of CA125 and EMAb were obviously degraded in the first to third months after surgery in test group and control group A(P < 0.05). There were no significant difference in the levels of CA125 and EMAb were at pre-operation,the first month,second month and third month of the post-operation between the test group and control group A(P > 0.05). There were no significant difference in the levels of CA125 and EMAb at pre-operation between the test group and control group B(P > 0.05). However, the level of EMAb was lower the first month after operation in the test group than that of control group B. In the follow-up, the adverse effects of the human interferon-α-2b were observed including low-grade fever, articular muscle soreness and gastrointestinal tract complaints. These appearances completely disappeared after drug withdrawal. It had less impact on the menstruation. Conclusion: The levels of CA125 and EMAb were similar in treatment of endometriosis with human IFN-α-2b and with Diphereline. Both of the treatments are better than the pure surgery. It has less impact on the menstruation and can avoid menostasia.

Objective:To compare two methods for the determination of compound cod-liver oil emulsion. Methods:The content of ciprofloxacin in compound cod-liver oil emulsion was determined by HPLC and UV, respectively. The determination by HPLC was performed on a Thermo C18 column (150 mm ×4.6 mm, 5 μm). The mobile phase was 0.025 mol·L-1phosphate-acetonitrile (87∶13)andpHwasadjustedto3.0±0.1withtriethylamine. Thedetectionwavelengthwas277nmandtheflowratewas1.5ml·min-1. The detection wavelength of UV was 277 nm. Results:The average recovery of HPLC and UV was 100. 44% and 100. 84% with RSD of 1. 01% and 1. 09% (n=9), respectively. The detection results of the two methods were compared by paired sample t-test, and no statistically significant difference was found (P>0. 05). Conclusion:The two methods are specific and accurate, and can be used for the determination of ciprofloxacin hydrochloride in compound cod-liver oil emulsion.

Objective:To optimize the extraction process of Hegan Lidan granules. Methods:In order to choose the optimal tech-nological parameters, the content of baicalin was determined by HPLC. An orthogonal method was utilized with solvent volume, extrac-tion time and extraction times as the impacting factors and the content of baicalin and extraction rate as the indices. Results:The opti-mal parameters were as follows:using 8-fold water as the solvent, the raw material was extracted three times with 2 h for each. Con-clusion:The process is steady and feasible, and can be used in the extraction of Hegan Lidan granules.

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

Objective To establish the quality standard of Qubizhitong gelata.Methods An HPLC method was used for the determination of paeonol with UV absorbance detection.The separation was performed using a Welchrom C18 with a mobile phase consisting of methanol-water (V∶V =45∶55).The UV detector operated at 274 nm.A TLC method was used for distinguishing Radix et Rhizoma Cynanchi Paniculati,Borneolum syntheticum and Curcumae Longae Rhizoma.Results and Conclusion The negative comparison displayed no disturbance.Linearity was within the range of 10 -320 μg/ml (r =0.9998).The average recovery was 97.42% and RSD was 1.01% (n =6).This method is suitable for quality control of Qubizhitong gelata.

Objective To study the therapeutic effect of recombinant adeno-associated virus carrying human endostatin gene therapy on endometriosis in mice model. Methods Recombinant adeno-associated virus vector carrying human endostatin gene and enhanced green fluorescent proteins gene (rAAV2-endostatin-EGFP) was constructed. Endometrium was from 12 patients with leiomyoma undergoing hysterectomy in Second Hospital, Tianjin Medical University between November and December 2008. Endometriosis models of nude mice were established by transplanting human endometrial fragments intooperitoneal surface. After 1 week, those 60 mice were divided into 3 groups: treatment group including 20 mice injected with rAAV2-endostatin-EGFP to ectopic lesion, control group including 20 mice injected with rAAV2-EGFP to ectopic lesion and blank control group including 20 mice injected with phosphate buffered saline (PBS) to the ectopic lesion. At 1, 2 and 3 weeks after treatment, those mice underwent laparotemy to observe the location and size of ectopic lesion in abdominal cavity. The expression of endostain protein, number of gland, microvessel density (MVD) and vascular endothelial growth factor (VEGF) were measured in ectopic lesions. The serum level of estradiol and progesterone were detected in nude mice among every groups. Results (1) All endometriosis of nude mice models were established successfully through peritoneum transplanting. After 1 week's treatment, flat lesion nodes, decreased gland number and narrow and atrophy glandular cavity were observed by light microscope. (2) The endostatin gene was transferred into nude mice successfully and expressed effectively. It was observed that endostatin protein expression was shown with enhanced green fluorescent proteins in ectopic lesion. (3) Glands number of ectopic lesion in rAAV2-endostatin-EGFP group(7.8±1.9,7.0±1.5 and 5.5±1.7) were significantly less than 10.1± 1.7, 10.2±2.0 and 9.8±2.4 in rAAV2-EGFP control group and 10.2±2.2,10.0±2.0 and 9.7±2.2 in PBS control group at 1,2 and 3 weeks after treatment(all P<0.05). Glands number of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (4) MVD of ectopic lesion in rAAV2-endostatin-EGFP group (12.2±1.5,11.4±2.1 and 9.0±1.4) was significantly less than those at rAAV2-EGFP control group (16.5±1.7,16.5±1.9 and 16.9±1.9) and PBS control group (16.2±1.6,16.0±1.6 and 16.3±1.7) at 1,2 and 3 weeks after treatment (all P<0.05) . MVD of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment(P<0.05). (5) The rate and density of VEGF expression at ectopic lesion in rAAV2-endostatin-EGFP group (35%, 30%, 25% and 1.60±0. 43,1.33± 0. 30,1.03±0.36) were significantly less than those at rAAV2-EGFP control group (80% ,75% ,85% and 2.43±0.53,2.43±0.29,2.66±0.45) and PBS control group (85% ,90% ,90% and 2.36±0.53,2.64± 0.57,2.53±0.52) at one 1, 2 and 3 ,weeks after treatment (all P<0.05). The expression of VEGF at ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (6) The level of estradial and progesterone in serum of nude mice of rAAV2-endostatin-EGFP group [ E_(2)> : (48±7 ) pmol/L, P: (61±8 ) nmol/L ] did not reach statistical difference when compared with those at rAAV2-EGFP control group [ E_(2): (50±9) pmol/L, P: (60±10) nmol/L] and PBS control group [E_(2):(48±7)pmol/L,P: (58±10)nmol/L,P>0.05]. Conclusions The recombinant adeno-asseciated virus carrying human endostatin gene therapy could inhibit angiogenesis at endometriotic lesions and not influence steroid level. The antiangiogenic gene therapy might become a novel option for endometriosis.