Objective To observe the attachment,shape,function and activity of human umbilical cord mesenchymal stem cells cultured on silk fibroin porous scaffolds in vitro,and to provide experimental foundation for the choice of scaffold in the study of adipocyte tissue engineering.Methods These silk fibroin porous scaffolds were subsequently seeded with hUCMSCs and cultured in vitro.The growth and function of hUCMSCs were observed and measured with fluorescence inverted microscopy,scanning electronic microscopy and MTT methed. Results hUCMSCs were fixed on silk fibroin porous scaffolds 1 or 2 days later,and multiplicative growth was observed on the 5th to 7th day.After about 10 days,the microholes of the scaffolds were overlayed by hUCMSCs.Scanning electronic microscopy and fluorescence inverted microscopy showed that cells adhered to scafold well and there was a lot of extra cellular matrix surrounding cells. Conclusion Silk fibroin porous scaffolds are ideal for attachment,growth,function maintenance and activity of hUCMSCs.and the scaffolds can be used as natural scaffolds for hUCMSCs in 3D culture.

Objective To detect the expression of miR-18a and estrogen receptor alpha (ER alpha) in single and multiple uterine flesh tumor tissues, discuss the relationship between miR-18a and ER alpha, and their effect in single and multiple uterine fibroids.Methods The expression of miR-18a and ER alpha in single and multiple uterine fibroids tissue paraffin section were detected by in situ hybridization method and immunohistochemical method, respectively.And the correlation between the miR-18a and ER alpha were evaluated.Results The expression of ER alpha in multiple uterine fibroids group was significantly higher than that of single uterine fibroids tissues (P<0.05);while miR-18a was weaker than that of single uterine fibroids tissues(P <0.05).The correlation results showed that miR-18a expression was correlated negatively with ER alpha expression either in single(r =-0.4421) and multiple uterine fibroids(r =-0.4181).Conclusion The expression of miR-18a is low in multiple uterine fibroids, while ER alpha had high expression.miR-18a could bea new target for the treatment of multiple uterine fibroids.

OBJECTIVE:To understand the application of proprietary Chinese medicine by clinicians in integrated medical insti-tutions,and provide reference for its rational use. METHODS:The questionnaire was designed,including related issues of proprie-tary Chinese medicine clinical application(such as medication basis of proprietary Chinese medicine,combined medication,clini-cal efficacy and its influencing factors,medication for special groups and so on)and suggestions;the questionnaires were delivered to the clinicians who worked in some second and third grade of integrated medical institutions located in southwestern regions and Chengdu,and the questionnaires was in the form of an anonymous;relevant data of available questionnaire were collected statisti-cally;the application situation of proprietary Chinese medicines and the existence question were analyzed,and suggestions were put forward. RESULTS:Totally 300 questionnaires were sent out,264 were effectively received,with effective rate of 88.0%. The clinicians involved in investigation were mainly in undergraduate,mostly the resident physicians and attending physician;only 6.8%of the physicians received the training of basic theory of traditional Chinese medicine after work;36.4%physicians often pre-scribe proprietary Chinese medicines,and the medication basis was mainly in package insert (64.0%) and clinical experience (55.3%);51.1% thought unclear indications affected the application of proprietary Chinese medicines,64.0% thought package in-sert can not meet clinical needs. In terms of answers in brief questions,the surveyed clinicians generally considered the package in-sert was too simple,which needed further perfected. CONCLUSIONS:The problems in clinical application of proprietary Chinese medicines should not be ignored. The relevant departments should strengthen the supervision of clinical application of proprietary Chinese medicines,and strengthen the re-evaluation of proprietary Chinese medicines market,as to provide data support for the continuous improvement of package insert. Medical institutions should carry out training timely for the physicians who prescribe pro-prietary Chinese medicines,the pharmacists should strengthen checking of proprietary Chinese medicines prescription and special prescription comment,and ensure prescription checking and comment play a active role in promoting rational drug use based on“proprietary Chinese medicines clinical application guiding principle”“prescription management method”.

Objective:To determine taurocholic acid (TCA) and taurodeoxycholic acid (TDCA) in artificial snake gall. Methods: HPLC was used in the quantitative analysis with spherisorb C 18 column, methyl alcohol 0.02 mol/L phosphric buffer (60∶40) as a mobile phase and detection wavelength at 210nm. Results: The linear range of TCA was from 0.28672 mg/mL to 2.8672 mg/mL, and the linear range of TDCA was from 0.26842 mg/mL to 2.6842 mg/mL. The average recoveries were 97.65% for TCA and 95.81% for TDCA. RSD were 0.79% and 3.27%, respectively. Conclusion: This method is simple and accurate.

OBJECTIVE:To provide reference for clinical rational use of anti-epileptic drugs. METHODS:In this retrospective review,serum concentrations of anti-epileptic drugs in a total of 499 patients who were treated with anti-epileptic drugs (such as sodium phenytoin,phenobarbital,carbamazepine and valproate sodium) in our hospital during 2007 were analyzed statistically. RESULTS:Among the patients receiving one kind of anti-epileptic drugs,206 cases (61.49%) were within normal range in blood concentration versus only 45 cases (44.12%) for patients receiving combined drugs. In addition,the above four drugs (sodium phenytoin,phenobarbital,carbamazepine and valproate sodium) were detected in 59.68% of the patients who took Chinese medicines,and among them,3.23% were within normal range in blood concentration. CONCLUSION:Monitoring on serum concentrations of anti-epileptic drugs is conducive to a better control of therapeutic concentration. It is advisable to refrain from using anti-epileptic drugs in combination but to adopt individualized administration. In addition,great importance should be attached to whether there are chemo-synthetic drug components contained in Chinese medicine.

Objective To assess the therapeutic effect of combination chemotherapy of gemcitabine and cisplatin by double way plus implantation of radioactive seed 125I implantation in treating stage Ⅲ non-small cell lung cancer. Methods Sixty cases with stage Ⅲ non-small cell lung cancer were randomly divided into two groups with random number table. In group A (in interventional treatment group, n = 30),the gemcitabine 1000 mg/m2 and one third of the cisplatin 100 mg/m2 was given using seldinger technique for transcatheter bronchial arterial infusion chemotherapy on day 1. Two-thirds of the cisplatin 100 mg/m2 was infused in veins on day 2 and 3. The gemcitabine 1000 mg/m2 was infused in veins on day 8, 21 days for a period. In group B (interventional - 125I groups), the method of combination chemotherapy of gemcitabine and cisplatin was the same as in Group A. After ten days of arterial perfusion, 125I seeds were implantated, 21 days for a period. All patients received at least 2 cycles. The imaging evaluation of patients after treatment standards included complete remission (CR), partial remission (PR), stable (SD),progressive disease (PD), effective rate (CR + PR)/30 and clinical benefit rate (CR + PR + SD)/30.Non-parametric rank sum test was used to compare short-term effect of the two groups treatment of two cycles.x2 test was used to compare year survival, Kaplan-Meier method was used to calculate median survival,log-rank test method was used to difference between the groups. Results In group A, there were 17 PR,9SD and 4 PD. The overall response rate was 56. 7% (17/30) and clinical beneficial rate was 86. 7% (26/30). In Group B, there were 2 CR, 21 PR, 7 SD. The overall response rate was 76.7% (23/30) and clinical beneficial rate was 100% (30/30). There was significant difference between the two groups (P =0. 036). In group A, the 1 year survival rate was 46. 7% (14/30) and the 2 year survival rate was 36. 7%(11/30), median survival time (MST) was 10 months . In group B, the 1 year survival rate was 76. 7%(23/30) and the 2 year survival rate was 63. 3% (19/30) , median survival time (MST) was 27 months.There was a significant difference between two group in 1 year survival rate (P = 0. 017), 2 year survival rate (P = 0. 039) and median survival time (P = 0. 006). Conclusion The treatment effects of Ⅲ stage non-small cell lung cancer by gemcitabine and cisplatin combination chemotherapy with double way plus radioactive seed 125I implantation was better than gemcitabine and cisplatin combination chemotherapy with double way.

Objective To assess the therapeutic outcomes of transcatheter arterial chemoembolization combined with percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis for primary hepatic carcinoma accompanied by portal vein tumor thrombus. Methods Thirty patients with primary hepatic carcinoma accompanied by portal vein tumor thrombosis of type Ⅱ and type Ⅲ were randomly divided into two groups. The Child-Pugh ratings (class A and B) of group A and B were 9 vs 9 (class A) and 5 vs 7 (class B) respectively (χ~2 = 0.201, P > 0.05). The constitution of Type Ⅱ and type Ⅲ portal vein tumor thrombus in group A and B were 8 vs 9 and 6 vs 7 respectively (χ~2 =0.002, P>0.05). The median values of ALT, TBIL, ALB and AFP in group A and B were 58.7U/L vs 70.5 U/L (W=191.5, P>0.05), 21.4 μmol/L vs 21.7μmol/L (W=203, P>0.05), 35.3 g/L vs 37.5 g/L (W = 214, P > 0.05) and 680 μg/L vs 873 μg/L (W = 179. 00, P > 0.05) respectively. Group A was treated with transcatheter arterial chemoembolization (TACE) using emulsion made up of adriamycin, cisplatin, mitomycin and ultraliquidlipiodol plus percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis using emulsion consisted of cisplatin and ultraliquidlipiadol, while group B was treated with TACE only as a control group. Survival analyses were performed via the Kaplan-Meier test in SPSS11.5 with the log-rank tests with an threshold of 0.05. Results The 3, 6 and 12 months survival cases of group A and B were 11 vs 10, 10 vs 3, and 7 vs 0 respectively. The median survival time of group A and group B were 14.0 months and 4.0 months respectively. The difference of the two groups was significantly (χ~2 =11.728, P<0.01). There was no severe side-effect related to therapy in both groups. Conclusion Comparing with the control group, TACE combined with percutaneous injection of chemoembolization agent intra-portal vein tumor thrombosis could significantly prolong the median survival time of patient with primary hepatic carcinoma accompanied by type Ⅱ and type Ⅲ portal vein tumor thrombosis.

Objective To observe the therapeutic effects of arsenic trioxide combined with transcatheter arterial chemoembolization on treatment of primary liver cancer with pulmonary metastases.Methods Sixty patients were randomly divided into two groups:group A (treatment group,n =30) and group B (control group,n =30).Group A was received periodic transcatheter arterial chemoembolization (TACE) and 10 mg arsenic trioxide by intravenous infusion for 5 hours per day,3 days after TACE.Each cycle consisted of 14 days' administration,and repeated after 2 weeks.Each patient was received 3-4 successive cycles.Group B was received periodic TACE alone.Objective efficiency,benefit rate,quality of life and the correlates with metastatic tumor size and number in the both groups were recorded.Results The objective efficiency was 26.7％ (8/30),and the benefit rate was 60.0％ (18/30) in group A,while they were 0 and 16.7％ (5/30) in group B with significant statistics differences (x2 =7.067,P =0.008;x2 =11.915,P =0.001).The quality of life was improved in 4 patients and stable in 18 of group A,while no patient was improved and 13 were stable in group B (x2 =9.669,P =0.008).There was a significantly positive correlation between the tumor burden and therapeutic effect (Kendall r =-0.765,P ＜ 0.001 ;Spearman r =-0.821,P ＜ 0.001).Conclusion Arsenic trioxide combined TACE is an effective treatment method in treating primary liver cancer with pulmonary metastases.

ObjectiveTo develop a cell culture system with consistent expression of whole hepatitis C virus (HCV) gene and Renilla luciferase gene and to facilitate the study on HCV pathogenesis and the screening of new antiviral drugs.MethodsRenilla luciferase (RLuc) reporter gene and a mutation that could yield higher virus gene expression were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Naǐve Huh7.5 cells were infected by the supernatant from the viral RNA transfected cells.HCV replication and infection were determined by virus titration,Renilla luciferase assay,immunofluorescence assay and western blotting.IFN-α was used to evaluate the feasibility of this system for anti-HCV new drug screening.ResultsThe viral RNA replicated efficiently in transfected cells.These cells could produce high titer of HCV-Rluc reporter virus and the virus titer reached to 1.5 × 104 FFU/ml at day 15 of posttransfection.The activity of Renilla luciferase was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-Rluc reporter virus.ConclusionThe recombinant HCV-JFH1-Rluc reporter gene system is sensitive and efficient.It can be a useful tool for high throughput screening of anti-HCV drugs.

Objective To develop a time saving and sensitive cell culture system based on hepatitis C virus chimera expressing enhanced green fluorescent protein(EGFP) and to facilitate the study on HCV pathogenesis and screening of anti-HCV drugs.Methods Enhanced green fluorescent protein reporter gene and a mutation V2440L that can yield higher virus titers were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Viral RNA in supernatant of HCV RNA-transfected cells was determined after transfection by RT-PCR.HCV replication and infection were determined by immunofluorescence assay.IFN-α was used to evaluate the feasibility of this system for anti-HCV drugs screening.Results The viral RNA replicated efficiently in transfected cells.These cells can produce HCV-EGFP reporter virus.Viral RNA levels in supernatant were 3.06× 105 copies/ml and 7.96×106 copies/ml at 72 h and 9 d after transfection,respectively.The virus titer reached to 104 FFU/ml 9 d after transfection.The expression of EGFP was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-EGFP reporter virus.Conclusion The recombinant HCV JFH1-EGFP reporter gene system is a time saving,cost effective and sensitive method for studying viral replication cycles and screening of anti-HCV drugs.