Objective To evaluate the effect of silencing leptin by small interfering RNA(siRNA)on the expression of lep‐tin ,and apoptosis ,proliferation and intracellular Ca2+ concentration([Ca2+ ]i )of hepatic stellate cells(HSCs)and to provide evi‐dence for liver fibrosis gene therapy.Methods HSCs were divided into normal control group ,blank vector group ,siRNA nega‐tive control group and leptin‐siRNA group.After transfection of the leptin‐siRNAs into HSCs ,cell proliferation was measured by MTT assay.Cell cycle and apoptosis were measured by flow cytometry.Expression of leptin was detected by immunocyto‐chemistry and Western blot. [Ca2+ ]i was measured by Fura‐2/AM loading.Results Compared with the normal control group , the blank vector group and the siRNA control group ,the protein expression of leptin and the cell growth were significantly in‐hibited in the leptin‐siRNA group(P<0.05). The proliferation rate of HSCs was significantly different at different time points (24 ,48 and 72 h)(P<0.05).The cell apoptosis rate was increased significantly in the leptin‐siRNA group(P<0.01).At the same time ,Leptin‐siRNA‐induced [Ca2+ ]i was also significantly reduced(P<0.05).Conclusion The leptin gene may play an important role in liver fibrosis progression and is potentially a novel predictive and prognostic marker for liver fibrosis.

Since HBV DNA integration was discovered for the first time in 1980, various methods have been used to detect and study it, such as Southern Blot, in situ hybridization, polymerase chain reaction and so on. HBV DNA integration is thought to be random on the whole although some hot spots of integration were described by some researchers, one of which might be the repetitive sequences of the genomic DNA. Besides, DNA damage, especially double-strand breaks could promote HBV DNA integration into host genome. HBV DNA integration into cells may damage the stability of the genome, cause DNA rearrangement, promote DNA deletion and induce the formation of HCC.

Objective:To study the mental state of primigravida before delivery and the effect of psychological help on their mood and delivery type.Method:272 primigravida who was just near their delivery were assessed with STAI and VAS for anxiety in Labor, 152 of them received psychological help including support from medical staff and family members in a sweet environment with accompanied family members?The other 120 primigravida received routine labor service.Result:During labor, primigravida had bad mood,psychological help deceased their anxiety and fear, and was helpful in upgrading the rate of delivery.Conclusion:Bad mood of primigravida is common, supportive attitude and environment are helpful to both mental state and their delivery.

Objective To construct an expression vector harboring CYP2B1 suicide gene, and detect its expressions in tumor cell lines. Methods PCR amplification was performed using primers based on murine CYP2B1 gene sequence from gene bank and pc3/2B1 as template. PCR product was directly inserted an eukaryotic expression plasmid pcDNA3.0. The recombinants were analyzed and identified by restriction enzyme analysis, PCR and sequencing. Then the recombinant vector pcDNA3.0/CYP2B1 was transfected into three tumor cell lines by liposome-mediated method. The expressions of CYP2B1 gene in all the cell lines were detected by RT-PCR method. Results pCDNA3.0/CYP2B1 vector was successfully constructed, and could express CYP2B1 mRNA in the three tumor cell lines. Conclusion Eukaryotic expression vector pcDNA3.0/CYP2B1 containing CYP2B1 gene under the control of a CMV promoter is an novel effective expression vector for tumor gene therapy.

Objective The newly developing gene therapy method and dominant negative mutants were bein g used as new promising HBV therapy method, and a dominant negative mutant of HB V X g ene we have reported in our previous report has some effects both on HBV replica tion and expression in transient expression, but the effects were interfered by persistent secretion of HBV in HepG 2 2.2.15 cell line in the experiment. To mak e sure the effects of dominant negative mutant of HBx gene, we established a HBx DN stably expressing cell clone, and evaluated the effects of HBx dominant negat ive mutant on HBV gene expression. Methods The prev HBx-GFP dominant mutant and the plasmids pRev Xwt, pRev GFP which contain the wild type X gene or GFP gene then transfected into HepG 2 2.2.15 cells by liposome. The HBsAg, HBeAg by in media were as sayed by RIA and HBV-related RNA were assayed by Northern blot. Results The pRev HBx-GFP, GFP and wild type X constructs can be effectively expressed in HepG 2 2.2.15 cells. The stable expressed HBx -GFP can significantly reduce HBeAg, HBeAg in media and the HBV-related RNA in HepG 2 2.2.15 cells, but not for pRev Xwt and pRev GFP. Conclusions The dominant negative mutant pRev HBx-GFP can significantly inhibit the HBV gen e expression. It also suggested that X gene may be one promising target for HBV gene therapy.

Amounts of Certain elements in Anopheles sinensis, both normal ones (emerged mosquitoes, d0, d5, d8, d12 and d18 after taking blood meal) and those infected with Brugla malayi microfilaria (d5, d 8, d12 and d18 after infection), were determined using IL-551 and PE-603 atomic absorption spectrometer(AAS). The results showed that 9 kinds of trace elements including Fe, Zn, Cu, Cd, Al, Pb, Mn. Ni and Cr and 4 kinds of macro elements as K, Na, Ca, Mg were present in both the noninfected and infected mosquitoes. Comparing the contents of the elements between the noninfected and infected mosquitoes, the amounts of many kinds of elements reduced obviously in mosquitoes infected with microfilaria. The quantity and kinds of elements reduced in mosquitoes with the days of infection, for instance, 10 kinds of elements on the 5th day, 11 on the 8th day, 9 on the 12th day and 7 on the 18th day after infection(Tables 1, 2).

Vitiligo is a kind of difficult to treat skin disease. Its pathogenesis is not very clear and the treatment is also difficult. In this paper, according to basic theories of traditional Chinese medicine (TCM) from basic drug selec-tion, visceral syndrome differentiation, harmonizing qi and blood, expelling wind and eliminating dampness, treatment according to four seasons, medication according to meridian pathways, psychotherapy and nursing. Experiences of professor W ang Jusheng in vitiligo treatment were introduced in order to provide a beneficial reference.

Objective To evaluate the diagnostic level of infantile vitamin B_1 deficiency encephalopathy.Methods Clinical featuresand CT images in 45 cases of infantile vitamin B_1 deficiency encephalopathy proved by clinical treatment and follow-up were analyzedretrospectively.Results CT images depicted bilateral symmetrical low-density regions in the lentiform nucleus and the head of caudatenucleus in all 45 cases.The regions were reniform,upsiloid,butterfly or oval.Bilateral symmetrical low-densities of frontal,temporal or parietal lobes were found in some cases.Other CT findings included extensions of longitudinal fissures,lateral fissures and cerebral grooves.Experimental treatments of vitamin B_1 made remarkable clinical improvements.Conclusion CT scanning is extremely useful in thediagnosis of infantile vitamin B_1 deficiency encephalopathy.

Objective To investigate the efficacy and prognostic factors in patients with stage Ⅰ b and Ⅱ a cervical carcinoma by radical radiotherapy.Methods Between January 1999 and January 2012,108 patients with stage Ⅰ b and Ⅱ a cervical carcinoma received radical radiotherapy in Cancer Hospital,Chinese Academy of Medical Sciences were included and analyzed retrospectively.Patients of stage Ⅰ b1,Ⅰ b2,Ⅱa1 and Ⅱa2 were 18 (16.7％,18/108),38 (35.2％,38/108),33 (30.6％,33/108) and 19 (17.6％,19/108),respectively.Results The 5-year overall survival rate was 76.2％ and the 5-year disease free survival rate was 75.6％.Totally 25 (23.1％,25/108) patients developed recurrent disease,16 of them (64％,16/25) had local recurrences,6 (24％,6/25) had distant metastases and 3 cases had both local recurrence and distant metastases.Among patients with recurrent disease,23 died and 2 survive with tumor.Totally 24 patients died,23 of them died due to tumor recurrence and the other one died of other reason.The univariate analysis showed that,lymph node metastasis,squamous cell carcinoma antigen (SCC) levels before treatment,SCC levels after treatment 1 month had relation with overall survival time in patients with stage Ⅰ b and Ⅱ a cervical carcinoma (all P ＜ 0.05).The multivariate analysis showed that,lymph node metastasis and SCC levels after treatment 1 month were the independent prognostic factors for overall survival time for the cervical squamous cell carcinoma (OR =2.5,4.4 ; all P ＜ 0.05).Conclusions By means of radical radiotherapy,stage Ⅰ b and Ⅱ a cervical carcinoma patients with lymph node metastasis and SCC levels≥ 1.5 mg/L after treatment one month have poor prognosis.While,stage Ⅰ b and Ⅱ a patients with concurrent chemoradiotherapy after neoadjuvant chemotherapy did not affect the prognosis.The 5-year survival rate with concurrent chemoradiotherapy was higher than that of radiotherapy.

Objective To find a novel operative modality with sphincter preservation in the treatment of middle and low rectal carcinoma. Methods Pull through lower resection was performed on 28 rectal cancer patients. The distance between the anal verge and the lower margin of the tumor was 6～8cm(20 patients) or 8～10cm(8 patients), including 8 patients in Dukes A stage, 16 Dukes B and 4 Dukes C. The resected line from tumor distal margin was 2cm, 3cm, and 4cm, respectively. Results There was no operative death, anastomotic fistula or anastomotic stenosis in these cases. Mean follow up period was 30 months. Local recurrence was found in two cases (7.1%) 18 months after the operation, and 26 cases were cancer free till the end of the follow up. Defecation was satisfactorily controlled 8～12 weeks after the operation. Conclusions Pull through Welch procedure could meet the criterion of the radical resection of rectal carcinoma,and keep the internal and external sphincter muscles intact in the superior lower anterior resection. The normal defecationcan can maintain after the operation due to the preservation of internal and external sphincter muscles.