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 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.

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).

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.

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 evaluate the outcomes and the prognostic factors for advanced squamous cervical cancer after concurrent chemoradiotherapy (CCRT).Methods Totally 172 patients with International Federation of Gynecology and Obstetrics stage Ⅱ b-Ⅳ who were treated in Cancer Hospital,Chinese Academy of Medical Sciences between January 2007 and December 2008 were retrospectively analyzed.Patients were received external radiotherapy,high-dose rate brachytherapy and cisplatin-based chemotherapy concurrently.Results The median follow-up period was 54.5 months.The 2-year and 5-year overall survival (OS) were separately 81.5％ and 68.8％.The 2-year and 5-year progress-free survival (PFS) were separately 69.2％ and 63.1％.Using univariate analysis followed with multivariate analysis,the results showed that these clinicopathological factors including stage (Ⅲ and above versus Ⅱ b; P =0.021,HR =1.95 ; P =0.020,HR =1.86),maximum diameter of local tumor size (＞4 versus ≤4 cm; P =0.009,HR =2.55 ; P =0.033,HR =1.94),squamous cell carcinoma antigen (SCC) level before treatment (＞3 versus ≤3 μg/L; P =0.010,HR =2.47; P =0.013,HR =2.09) and retroperitoneal lymph node status on imaging (para-aortic lymph node positive versus negative,P =0.009,HR =3.00,P=0.010,HR =2.74; pelvic lymph node positive only versus negative,P =0.044,HR =1.98,P =0.033,HR =1.92) had the significant effect on OS and PFS.Patients with no above adverse prognostic factor were assigned to Group A (n =18),those with one factor were assigned to Group B (n =43),and those with no less than two factors were assigned to Group C (n =1 11).Among three groups,the 2-year OS were separately 94.1％,97.7％ and 73.1％,the 5-year OS were separately 81.4％,90.1％ and 58.6％,the 2-year PFS were separately 88.2％,90.4％ and 57.9％,the 5-year PFS were 82.4％,87.9％ and 50.0％.The results showed that group C was significant difference from Group A or B in OS and PFS (all P ＜ 0.05),while Group A had no significant difference from Group B in OS and PFS (P ＞ 0.05).Conclusions Stage Ⅲ or above,maximum diameter of local tumor size ＞ 4 cm,SCC level ＞ 3 μg/L before treatment and positive retroperitoneal lymph nodes on imaging are four independent adverse factors for prognosis of squamous cervical cancer of advanced stage after CCRT.The treatment of patients with no less than two adverse factors should be considered to be improved.

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.