Objective:To study the feasibility of treating endometrial carcinoma by adenovirus vector containing wild type PTEN gene(Ad-PTEN) . Methods:Twenty-four BALB/c mice with endometrial carcinoma were equally divided into 3 groups,with Ad-CMV transfected or untransfected mice as control group.The tumorigenic ability of implanted endometrial carcinoma was observed after ex vivo transfection with Ad-PTEN.LacZ was used as a reporter to determine the efficiency of adenovirus gene transfer.Fifteen BALB/c nude mice with endometrial carcinoma were equally divided into PBS control,Ad-CMV group and Ad-PTEN group.When the implanted tumors grew to 4-5 mm in diameter,5?10~8 pfu/100 ?l Ad-PTEN,Ad-CMV or PBS were injected intratumorally once the other day for 3 times.The changes of tumor size and the presence of adverse effects were observed.Fifteen days after treatment all mice were sacrificed and tumors were excised for routine histological examination.(Results:)Ad-PTEN transfected endometrial carcinoma cell line RL 95-2 completely lost tumorigenic ability in BALB/c mice.The tumorigenic rates were 0,100% and 100% in Ad-PTEN,Ad-CMV and PBS control group,respectively.The in vivo efficiency of adenovirus mediated gene transfer into endometrial carcinoma was 80% 96 h after Ad-CMV-LacZ injection.The growth of tumor in Ad-PTEN group was obviously decreased and the tumors' volumes in Ad-PTEN group were obviously smaller than those in Ad-CMV and PBS group(P

Objective:To observe the expression of Ad-PTEN in human endometrial carcinoma cell line RL95-2 after in vitro infection and investigate the mechanism by which Ad-PTEN inhibits tumor cell proliferation and induces apoptosis. Methods: Ad-PTEN was constructed through a bacterial homologous recombinant system; the expression of Ad-PTEN in RL95-2 cells was determined by RT-PCR, Western blotting and immunohistochemical staining. The efficiency of adenovirus mediated gene transfer of Ad-PTEN was determined by X-gal staining. The inhibitive effect of Ad-PTEN on RL95-2 cells proliferation was determined by cell growth analysis and MTT assay; the morphologic and ultrastructural changes of RL95-2 cells transfected with Ad-PTEN were observed by the light and electron microscopy; and Flow cytometry was used to study the cell cycle and apoptosis. Results: The expression of Ad-PTEN mRNA and protein in RL95-2 cells was confirmed by RT-PCR, Western blot and cell immunohistochemical staining. The efficiency of adenovirus mediated Ad-PTEN gene transfer was 100% when the multiplicities of infection (MOI) was 50. Exogenous PTEN gene significantly suppressed the growth of RL95-2 cells and iduced the apoptosis. Ad-PTEN could also induce cell cycle arrest (G_(0)/G_(1) ) and activated caspase-3 . Conclusion: The constructed Ad-PTEN transfection system is highly efficient in introducing wild type PTEN gene into human endometrial carcinoma RL95-2 cells. Ad-PTEN can strongly inhibit cell proliferation and induce apoptosis in RL95-2 cells, which may be associated with cell cycle arrest (G0/G_(1)) and the activation of caspase-3.

Histone methyltransferase modulates heterochromatin formation,genomic imprinting and genetic transcription by regulating the combination of histones and DNA.As encoding genes of histone methyltransferase,mixed-lineage leukemia (MLL) genes are found to have close relationship with tumors.Recently,MLL2,a member of this family,has been found highly expressed anomalously in breast cancer,colorectal cancer and lymphoma.Whether MLL2 participates in the progression of cancer,the time phase of its participation and its specific role are still remained to further study.

Objective To evaluate clinical effects of combined use of hysteroscopy and laparoscopy in the diagnosis and treatment of infertility with uterine septum,and to analyze the relationship between infertility and uterine septum. Methods Surgery using hysteroscopy combined with laparoscopy was performed in 110 patients with infertility accompanying uterine septum,including primary infertility in 78 patients and secondary infertility in 32 patients.The relative infertility factors and post-operative pregnancy prognoses were analyzed. Results Unexplained infertility accounted for 40% of patients(44/110).As of March 2006,a total of 82 patients were successfully followed.The total post-operative pregnancy rate was 45.1%(37/82),consisting of 46.6% in patients with primary infertility(27/58) and 41.7% in patients with secondary infertility(10/24),without significant difference between the two groups(?~2=0.164,P=0.686).The post-operative pregnancy rates in patients with unexplained infertility and etiologically-clarified infertility were 51.2%(22/43) and 38.5%(15/39),respectively,without significant difference(?~2=1.332,P=0.248).In patients with unexplained infertility,the post-operative pregnancy rate for primary infertility was 56.7%(17/30) and for secondary infertility,38.5%(5/13). Conclusions Combined use of hysteroscopy and laparoscopy for infertility with uterine septum can improve the post-operative pregnancy rate.The presence of uterine septum bears some relationship to the incidence of infertility.

Objective To discuss the influence of gestational hypothyroxinemia to the pregnancy outcomes and fetus development,and find the evidence of hormone replacement therapy.Methods The clinical data of 1141 gravida admitted from Nov.2014 to Oct.2015 were retrospectively analyzed,including the data of systematic antenatal examination,all the data of pregnancy,the materials of delivery,the last ultrasound examination,production status and the thyroid stimulating hormone (TSH) of the newborn etc.,to find the difference of related index.Results Of the 1141 gravida with integral data,200 had past history of thyroid disease,189 showed below normal of free thyroxine (FT4) and 752 were normal ones.The 189 gravida with normal TSH but lower FT4 were divided into group A (0-5％ lower than the normal FT4 value,n=60),group B (5％-10％ lower than the normal FT4 value,n=40) and group C (10％ and above lower than the normal FT4 value,n=89).The ones with both normal TSH and FT4 value served as control group.Compared to the control group,the higher premature delivery rate,incidence of gestational diabetes mellitus and cesarean delivery rate (P＜0.05) were found in group C,and more gravida in group B had a history of hypertension and dyslipidemia during pregnancy (P＜0.05).The cesarean delivery rate of group B and C were higher than group A.Meanwhile,the rate of group B was higher than control group (P＞0.05).At delivery,the maternal weight,BMI,diastolic pressure,and head circumference of fetus in the last ultrasound examination were higher in group C than in control group (P＜0.01),but the gestational weeks of the newborn were shorter in group C (38.55 ± 1.86 weeks) than in control group (39.14 ± 1.57 weeks,P＜0.01).The 189 gravida with lower FT4 were divided into two groups according to the thyroid peroxidase antibody (TPOAb) level.The head circumference of fetus in the last ultrasound examination was higher in TPOAb(+) group than in TPOAb(-) group (45.99 ± 62.36cm vs.33.23 ± 2.08cm,P＜0.01).Conclusions The influence of gestational hypothyroxinemia to pregnancy outcomes and fetus development cannot be ignored,especially for the pregnant women with lower FT4 value (10％ and above lower than the normal) or with positive TPOAb.It is suggested to take the thyroid function test in the early stage of pregnancy for those pregnant women mentioned above.

BACKGROUND: Insulin like growth factor 2 (IGF2) is an important embryonic mitosis growth promoting factor, whichplays a critical role in the process of maintaining normal cell growth. The gene expression of IGF2 is under epigeneticregulation in the way of genomic imprinting. Imprinting loss of IGF2 is likely to be associated with the abnormaldevelopment of the individual and tumorigenesis.OBJECTIVE: To investigate the effect of imprinting loss of IGF2 gene on the differentiation of mouse embryonic stemcells into islet-like cells.METHODS: Two kinds of mouse embryonic stem cells (SF1-G and SF1-1) were induced to differentiate into islet-likecells in vitro. The expression of Insulin gene was detected by real-time PCR and cell immunofluorescence. Theexpression of IGF2 was detected by the polymerase chain reaction-restriction fragment length polymorphism in the cellsbefore and after induced differentiation. The level of insulin released at terminal differentiation stage was tested by insulinrelease assay in vitro.RESULTS AND CONCLUSION: (1) There was no change in the imprinting state of the two kinds of mouse embryonicstem cells with normal and imprinted IGF-2 gene before and after differentiation. (2) In the induced cells, the expressionlevel of insulin in the SF1-1 group was higher than that in the SF1-G group, although there was no significant differencein the insulin release between the two kinds of cells. (3)The imprinting loss of IGF-2 gene may be related to theup-regulation of insulin mRNA expression in terminal cells during induced differentiation.

Aim To compare the effects of the non-peptide angiotensin Ⅱ receptor type Ⅰ antagonist,Losartan,and the active vascular peptide,calcitonin gene-related peptide(CGRP),on the proliferation of vascular smooth muscle cells induced by angiotensin Ⅱ,and to explore the mechanism of depressor effect of Losartan and CGRP in vivo.Methods MTT,Thymidine incorporation and flow cytometry,were used to determine the ability of proliferation of VSMC induced by angiotensin Ⅱ in the presence or absence of Losartan or CGRP,Western blotting was used to determine the activity of ERK1/2.Results Losartan or CGRP inhibited the viability,DNA synthesis,cell proliferation index,and the activity of ERK1/2 in a dose-dependent manner.Conclusion Losartan or CGRP significantly inhibits the proliferation of VSMC induced by angiotensin Ⅱ;the inhibitory effect of CGRP is stronger than that of Losartan.The signaling path way is involved in ERK1/2.

Objective:To explore the way to separate and culture eutopic and ectopic endometrial glandular cells and their stromal cells,providing an in vitro cell model of endometriosis for study of its mechanism. Methods:Digestion,filtration and sedimentation were used to isolate and culture the eutopic and ectopic endometrial glandular cells and their stromal cells. The estrogen level was imatated to study the way of promoting cell growth. Morphological characters of eutopic and ectopic endometrial cells were examined using optical microscope. Results:The success rate of separation and culture of normal control endometrial glandular cells and its stromal cells was 91.7%(11/12);of eutopic endometrial glandular cells and its stromal cells of endometriosis was 93.8%(15/16);of etopic endometrial glandular cells and its stromal cells of endometriosis was 75.0%(12/16) . Conclusion:The cultured eutopic and ectopic endometrial cells is more like human body feature than the endometriosis model of animals. So the isolation and culture of eutopic and ectopic endometrial glandular cells and their stromal cells may serve as an in vitro experimental model.

Objective:This paper aimed to investigate the effects of isocorydinone on cell proliferation in SiHa human cervical carcinoma cell lines. Methods:Different concentrations of isocorydione (100, 200, 400, 800, and 1200 μmol/L) were used to treat SiHa human cervical carcinoma cells in vitro for 24, 48, and 72 h. Methyl thiazolyl tetrazolium (MTT) assays were conducted to determine the inhibitory action of isocorydione. Flow cytometry was performed to detect the cell cycle in SiHa human cervical carcinoma cells af-ter treatment with 400 μmol/L isocorydione. Hoechst 33342 staining was used to observe the micro-morphological changes of SiHa cell nucleus after the treatment. The expression of Bcl-2, Bax, and caspase-3 proteins in cervical carcinoma SiHa cell lines was determined using western blot analysis. Results: MTT assays showed that isocorydione inhibits the proliferation of SiHa cells in a dose- and time-dependent manner (P<0.05). The flow cytometry results showed that SiHa cervical carcinoma cells treated with different concen-trations of isocorydione exhibited increased cell cycle. Compared with the control group, Hoechst 33342 staining showed that SiHa cells became narrow, with nuclear pyknosis and fragmentation, and formed an apoptotic body after treatment with 400 μmol/L isocoryd-ione for 48 h. Furthermore, western blot analysis proved that isocorydione significantly inhibited the proliferation of SiHa cell lines, and the expression of Bax protein was increased. By contrast, the expression of Bcl-2 protein decreased gradually. Consequently, the ra-tio of Bax/Bcl-2 increased, as well as the expression of caspase-3 protein. Conclusion:Isocorydione exhibited an overt inhibitory ac-tion on SiHa cells. Isocorydione promoted the occurrence of cell apoptosis, which may be associated with related proteins of mitochon-drial apoptotic pathway.

Objective To investigate the efficacy of escitalopram combined with psychoanalysis in the treatment of refractory depression. Methods A total of 63 patients were randomly divided into escitalopram group ( n = 32) and escitalopram combined with psychoanalysis group( n = 31 ). All patients were evaluated with Hamilton depression Rating Scale(HAMD). Results After treatment,the scores of HAMD in two groups were both significantly lower than those before treatment. In the 8th ( ( HAMD ( 17.35 ± 2.98 ) ), 12th ( ( HAMD (9. 26 ±3.46) )weekend of treatment, the scores of HAMD in study group were significanlly lower than those in control group(8 th:21.97 ± 3.26; 12 th: 15.28 ± 3. 18 ). There were no significant differences in side effects between study group and control group. Conclusion Escitalopram angumented with psychoanalysis takes effects better than escitalopram single and doesn't increase side effects in the treatment of refractory depression.