Objective To investigate the safety and efficiency of radiofrequency ablation (RFA) in the treatment of complicated multifetal gestations.Methods There were 6 multifetal pregnant women (gestational age ranged from 14+6 to 27 +2 weeks) diagnosed in the Department of Obstetrics,Provincial Hospital Affiliated to Shandong University:two with twin-twin transfusion syndrome (TTTS) stage Ⅳ,one with reversed arterial perfusion sequence,one with dichorionic triamniotic triplets,one with absence of a lower limb,one with severe intrauterine growth restriction.All of them accepted ultrasound-guided selective fetocide by RFA.Results (1) Blood flow of three reduced fetuses stopped completely after one RFA circulation,whereas the other three stopped after two circulations.One reduced fetus stopped heartbeating in 10 minutes after RFA; three reduced fetuses' heartbeats slowed down and stopped completely in 35 minutes after RFA ; and the heartbeats of the other two cases stopped completely within 3 to 7 minutes after RFA.The heartbeats of the reserved fetuses were normal.All of the operations succeeded.(2) The reserved fetuses received a series of ultrasound examinations after the operations.In Case 1,the ascites of the reserved fetus,which was 4.0 cm× 2.3 cm before RFA,disappeared two weeks later; and the umbilical artery systolic/diastolic (S/D) ratio,which was 3.35 before the operation,decreased to 2.70 six weeks later.Amniotic fluid depth decreased from 44.6 cm to normal two weeks after RFA.The reserved fetus received brain MRI three weeks after RFA and no abnormality was detected.In Case 2,the increased heart size (cardiothoracic ratio ＞ 0.35) of the reserved fetus recovered to normal size ten days after the operation ; and the umbilical artery S/D decreased from 4.69 to 3.39 seven days after the operation.Reserved fetuses of the other three cases were normal on ultrasound and MRI after the operations.In Case 6,the ascites of the reserved fetus,which was 2.3 cm × 1.5 cm before RFA,disappeared sixteen days after the operation.The brain MRI suggested normal three weeks after the procedure.Amniotic fluid depth reduced from 11.0 cm to normal two weeks after the operation.(3) Three women delivered normal premature babies,and the other three got healthy mature infants.At present,all children are still in follow-up,and their physical examinations suggest normal.Conclusions RFA is a safe,efficient,minimal invasive treatment,which provides a new choice for fetocide,especially for complicated monochorionic multifetal gestations.Fetocide by RFA can effectively improve the life quality of the reserved fetuses.

Objective To evaluate the clinical effect of different doses of leuprorelin acetate in in vitro fertilization-embryo transfer(IVF-ET).Methods From January 2011 to December 2011,the data of 268 patients undergoing IVF and (or) intracytoplasmic sperm injection (ICSI) in Reproductive Medical Center,Clinical College of PLA,Anhui Medical University were studied retrospectively.All the patients were divided into three groups based on with long protocol and controlled ovarian stimulation (COH) including 83cycles with 1.25 mg of leuprorelin in low dose group,68 cycles with 1.88 mg of leuprorelin in high dose group,117 cycles with 1.25 mg of diphereline in control group.The serum follicle stimulating hormone (FSH),luteinizing hormone (LH),estradiol (E2) and progesterone (P) before gonadotropin (Gn)administration on the days 3-5 of the menstrual cycle and on the day of hCG administration were detected,the dose and duration of Gn,number of oocytes retrieved,number of mature oocytes,the rates of fertilization,embryo cleaved,good-quality embryos clinical pregnancy and early miscarriage were compared among three groups.Results There were no significant differences in age,the level of LH and P on the day of hCG administration among three groups (P ＞ 0.05).The level of FSH was (3.8 ± 1.6) U/L in low dose leuprorelin group,(3.1 ± 1.4) U/L in high dose of leuprorelin group and (2.4 ± 1.3) U/L in diphereline group before Gn administration,which reached statistical difference (P ＜ 0.05).The mean length of Gn stimulation were (9.8 ± 1.7) days in low dose leuprorelin group,(10.5 ± 1.8) days in high dose of leuprorelin group and (11.1 ± 1.4) davs in diphereline group,which reached statistical difference (P ＜0.05).The mean dose of Gn was (24 ± 7) in low dose of leuprorelin group,which was significantly higher than (27 ± 9) in high dose of leuprorelin group and (28 ± 7) in diphereline group (P ＜ 0.05).The level of LH was (2.7 ± 1.6) U/L in low dose of leuprorelin group and (2.2 ± 1.0) U/L in diphereline group before Gn administration,which reached statistical difference(P ＜ 0.05).The cancel cycles were 5 in low dose of leuprorelin group,4 in high dose of leuprorelin group and 7 in diphereline group.The number of ovum was (14 ±7) low dose of leuprorelin group,(13 ±6) in high dose of leuprorelin group,(14 ±6) in diphereline group.The rates of fertilization was 66.26％ (758/1144)in low dose of leuprorelin group,67.01％ (589/879) in high dose of leuprorelin group and 68.54％ (1111/1621) in diphereline group,the rates of goodquality embryos was 64.22％ (472/735) in low dose of leuprorelin group,60.50％ (340/562) in high dose of leuprorelin group and 59.59％ (640/1074) in diphereline group,clinical pregnancy was 49％ (38/78) in low dose of leuprorelin group,42％ (27/64) in high dose of leuprorelin group and 50％ (55/110) in diphereline group,early miscarriage was 18％ (7/38) in low dose of leuprorelin group,15％ (4/27) in high dose of leuprorelin group and 15％ (8/55) in diphereline group,which did not show significant differences (P ＞0.05).Conclusions Both 1.25 mg and 1.88 mg leuprorelin acetate could obtain good downregulation effect and clinical outcomes.1.25 mg leuprorelin acetate could decrease patient's costs by reducing Gn dose and duration.

Objective To study the pathologic characteristics of eutopic endometrium in patients with endometriosis.Methods Pathologic characteristics of eutopic endometrium were studied in 176 patients with endometriosis in Peking Union Medical College Hospital from January 2007 to December 2008 retrospectively.Results About 72.2％(127/176)of eutopic endometrium were in proliferative phase,19.9％(35/176)of were observed as endometrial polyp,including 32 cases with simple endometrial polyp and 3 cases with abnormal hyperplasia combined with endometrial polyp.And 4.0％(7/176)showed abnormal hyperplasia.The incidence of pathologic changes in eutopic endometrium was 22.2％(39/176).Among 53 endometriosis patients combined with infertility,the incidence of pathologic changes of eutopic endometrium was 35.9％(19/53),which was significantly higher than 16.3％ in non-infertile patients (x2 =8.24,P =0.004).Among 65 cases with irregular menstruation,the incidence of endometrial polypus and endometrial hyperplasia were 20.0％(13/65)and 10.8％(7/65),which were significantly higher than 17.1％(19/111)and 0 in normal menstruation patients(x2 =13.839,P =0.003).Conclusions The eutopic endometrium of endometriosis were in proliferative phase state.The pathologic changes of eutopic endometrium were more in patients combined with infertility and irregular menstruation.

Objective To investigate the value of detection of fetal cell-free fetal DNA(cff-DNA)in maternal plasma in the prenatal diagnosis of chromosomal abnormalities.Methods The plasma from 3200 gravidas(singleton with 20.3 ± 3.8 gestational weeks)was collected from April 1st 2011 to May 30th 2012.They were divided into 3 groups:(1)To tally 1720 cases were included in the high-risk serological screening group,in which women were younger than 35 years and got high-risk results in serological screening;(2)To tally 1310 cases were included in the advanced age group,in which women's age was more than 35 years;(3)To tally 170 cases were included in the supplementary group,in which women were younger than 35 years and got low-risk results in serological screening,or women who didn't take serological screening tests.All the 3030 gravidas in group 1 and 2 didn't take invasive prenatal diagnosis because of fear of abortion or short of prenatal diagnosis.Cff-DNA were detected by next generation sequencing in Shenzhen BGI Genomics Center for clinical laboratory.Amniocentesis and karyotype analysis were provided to the positive cases and women with negative results were followed-up by telephone.Results(1)The 3200 cases took cff-DNA detection,and 31 cases got positive results,including 27 cases of trisomy 21 and 4 cases of trisomy 18.Sixteen cases of trisomy 21 and 1 case of trisomy 18 were in the high-risk serological screening group.7 cases of trisomy 21 and 2 cases of trisomy 18 were in the advanced age group.Four cases of trisomy 21 and 1 case of trisomy 18 were in the supplementary group.(2)And the 84％(26/31)cff-DNA detecting positive cases received amniocentesis.In the 27 trisomy 21 positive cases,23 received amnioeentesis and got karyotype of 47XN,+ 21,with the diagnostic accordance rate of 100％.In the 4 cases who didn't take karyotype analysis,fetal anomaly(ventricular septal defect,dextrocardia and choroid plexus cyst)was found in 1 case before 20 gestational weeks;intrauterine fetal demise happened in 1 case before getting the result;2 other cases who already had healthy children took abortion in the local hospital without taking amniocentesis.In the 4 trisomy 18 positive cases,3 took amniocentesis,2 of which were trisomy 18 and took abortion,the other was chimera(46,XN/47,XN,+ 18)with only 2％ cells of trisomy 18,with no malformation found after delivery.Hypoevolutism(3 weeks less than gestational week),general hydropsy and intrauterine fetal demise happened before the other case took amniocentesis.(3)Follow up of cff-DNA negative cases:until May 30th 2012,no Down's baby was found in the 1230 cases with cff-DNA test negative results.Conclusions(1)The non-invasive fetal trisomy test(NIFTY)by next generation sequencing is a safe,accurate and high throughput method for the prenatal diagnosis of trisomy-21.(2)Use NIFTY as a further screening for pregnant women with high-risk serological screening results could lower invasive prenatal diagnosis rate.(3)Cases with positive NIFTY test results should receive amniocentesis and karyotype analysis to confirm the diagnosis before abortion.

Objective To investigate the clinical value of non-invasive prenatal diagnosis using cell free fetal DNA(cff-DNA)in maternal blood.Methods From Sep.2010 to Mar.2012,103 pregnant women who came to Henan Province People's Hospital in the first trimestcr for prenatal diagnosis of scx-linked inherited diseases were included in the first trimester group.From Oct.2010 to Jan.2012,205 pregnant women undergoing amniotic fluid sampling for fetal karyotype analysis in the same hospital were included in the second trimester group.Real time quantitative PCR and fluorescent PCR were used to detect sex determining region of Y chromosome gene(SRY)and amelogenin gene(AML)on cff-DNA of the first trimester group.Moreover,12 Y chromosome STR loci analysis were performed for 33 male fetuses and their fathers.Massively Parallel Signature Sequencing(MPSS)was used for aneuploidy analysis in cff-DNA of the second trimester group.Results(1)In the first trimester group,there were 53 SRY positive and 50 SRY negative.Compared with the results of cff-DNA of chorionic villus samples,there was one SRY false positive and one false negative results,with a sensitivity of 98％ and specificity of 98％.For the AML gene test,there were two PCR products of male fetuses:102 bp fragment originating from X chromosome(AML X)and 108 bp fragment from Y chromosome(AML Y);but only AML X was found in products from female fetuses.In the first trimester group,102 bp and 108 bp fragments were detected in 52 cases,and only 102 bp fragment was found in the other cases.Compared to AML results from chorionic villus samples,there were 2 false negative results,with a sensitivity of 96％ and specificity of 100％.(2)For cff-DNA with plasma SRY over 30 copy/ml,Y STR loci were analyzed on cff-DNA of 33 fetuses and their fathers.The Y STR loci less then 200 bp were successfully detected,while Y STR loci with PCR products between 200-300 bp showed low signal or could not be amplicated;and no PCR products more than 300 bp were detected from cff-DNA.Comparing the detected Y STR loci of cff-DNA to the fathers,32 fetuses were concordant with their fathers'.Exogenous contamination was found in the rest one sample.(3)In the second trimester group,6 fetuses with abnormal karyotype(two trisomy 21,three trisomy 18 and one 45,XO)were detected by cff-DNA and were proved by karyotype analysis.Moreover,the MPSS results of cff-DNA revealed one 45,Y and one trisomy 16 whose karyotype analysis showed normal results.And in one case,MPSS suggested less chrX or chrY,that was proved to be 47,XYY by karyotype analysis.Conclusions(1)Cff-DNA in maternal blood can be used to determine fetal gender in early prenancy with considerable sensitivity and specificity.But the trace cff-DNA and the high maternal DNA background might have impact on the result.(2)Analysis of cff-DNA in maternal blood of the second trimester women showed that MPSS could be used for prenatal screening of trisomy 21 and trisomy 18.However,further research should be done for other chromosomes aneuploidy detection.

Objective To investigate the expression pattern and significance of two importantoocyte-secreted factors:growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)during oocyte maturation in women with polycystic ovary syndrome(PCOS)and infertile women due to husband factors.Methods Total of 25 oocytes[9 at germinal vesicle GV stage,9 at M Ⅰ stage and 7 at M Ⅱ stage]were obtained from 12 patients with PCOS and 82 oocytes(29 at GV stage,26 at M Ⅰ stage and 27 at M Ⅱ stage)were from 56 controls.The nested quantitative real time(RT)PCR was uscd to detect the abundance of GDF9 and BMP15 mRNA in each oocyte.Results(1)The expression level of GDF9 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 44.8(4.2-529.0),27.6(9.8-172.7)and 49.0(0.2-65.9)respectively,the expression in were 149.9(55.4-387.9),29.9(2.5-205.8)and 657.8(149.4-1376.2)in control group,respectively.The expression of GDF9 mRNA at M Ⅱ stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences didn't reach statistical significant at GV or M Ⅰ stage between the two groups(P ＞ 0.05).The expression of GDF9 mRNA displayed some changes at different maturation stage in controls(P ＜ 0.05,P ＜ 0.01),however,the expression didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).(2)The expression level BMP15 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 0.1(0.1-22.0),3.2(0.6-55.0)and 6.4(3.2-8.5),respectively,the expression were 41.6(6.5-96.1),4.0(2.0-10.4)and 49.7(2.3-139.5)in control group,respectively.The expression of BMP15 mRNA at GV stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences were not significant at M Ⅰ or M Ⅱ stage between the two groups(P ＞0.05).The expression of BMP15 mRNA also displayed some changes at different maturation stage in controls(P ＜ 0.05),however,the level didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).Conclusion It was suggested that the low expression of oocyte secreted factors in mature oocytes from PCOS patients might be associated with impaired oocyte quality and developmental competence in PCOS.

Objective To investigate factors with pelvic adhesions and the effect of different degrees pelvic adhesions on fallopian tube recanalization in infertile patients.Methods Total of 527 infertile patients undergoing hysteroscopy and laparoscopic surgery in Affiliated Hospital of Chinese People's Armed Police Forccs Logistics College were studied retrospectively.According to the extent of pelvic adhesions,tubal umbrella adhesions and atresia,377 cases were classified into adhesion groups,including 73 cases in grade Ⅰ,221 cases in grade Ⅱ,75 cases in grade Ⅲ and 8 cases in grade Ⅳ based on adhesion score.The 150 cases with no obvious pelvic adhesion were matched as control group.Among 8 cases with grade Ⅳ ahesion were exluded from ahesion group the relationship between pelvic adhesions and related history,abdominal lesions,tubal patency and the prognosis were studied.Results(1)Related factors:the frequency of pelvic adhesion and more than 7 years of infertility of 23.9％(88/369)in adhesion group were significantly higher than 12.0％(18/150)in control groups.(2)History:compared with the control group(12.7 ％,19/150;28.7％,43/150;11.3％,17/150;12.0％,18/150;17.3％,26/150),patients with pelvic adhesions present more incidence abortion(23.6％,87/369),uterine cavity operation(38.2％,141/369),ectopic pregnancy(20.9％,77/369),pelvic inflammatory disease(25.5％,94/369)and abdominopelvic surgery (31.4％,116/369).(3)Endoscopy exploration:the incidence of hydrosalpinx(24.7％,91/369),tube distorted(15.7％,58/369)and salpingostomy(72.9％,269/369)in adhesion group were higher than those in control group(2.0％,3/150;4.0％,6/150;12.0％,18/150),but relatively lower incidence of pelvic endometriosis lesions(5.7％,21/369)and mesosalpinx cysts(16.3％,60/369)than those in control group(16.0％,24/150;30.0％,45/150).The rate of proximal tubal recanalization(59.5％,91/153)in adhesion group was lower than 75.4％(52/69)in control group.However,the rate of distant tubal recanalization of 84.4％,(281/333)in adhesion group and;13/15 in control group didn't show statistical difference.(4)Prognosis:the rate of ectopic pregnancy of 9.7％(29/299)in adhesion group was significantly higher than 3.1％(4/128)in control group.Among cases with grade Ⅲ adhesion exhibited the highest rate of ectopic pregnancy(13.0％,7/54;OR =4.62,95％ CI:1.29-16.50).(5)Multivariate analysis:it was found that more than two drug abortions(OR =3.29,95％ CI:1.34-8.07),pelvic and (or)abdominal surgery history(OR =2.20,95％ CI:1.35-3.57)and pelvic inflammatory disease history (OR =1.54,95％ CI:1.21-1.97)were risk factors with pelvic adhesions.Conclusion More than or equal to two drug abortion history,pelvic inflammatory disease and pelvic and abdominal surgery damage were important factors for pelvic adhesions of infertility patients,which may decrease the possibility of proximal tubal recanalization and increase ectopic pregnancy risk.

Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.

Objective To investigate the expression of insulin receptor isoforms(IR,including IR-A and IR-B)in endometrial carcinoma(EC),and explore the role of IR-A in the growth of endometrial carcinoma cells.Methods The expression of IR isoforms were detected by reverse transcription(RT)-PCR and real-time PCR in 4 different endometrial cancer cell lines(HEC-1-A,Ishikawa,RL95-2 and KLE),with human breast cancer cell line MCF-7 cells and hepatocellular carcinoma cell line Hep-G2 as positive control and in the endometrial cancer tissue specimens of 42 cases,admitted in Peking University People's Hospital from November 2007 to July 2009,with normal endometrial tissues from 15 cases of ovarian neoplasms at the same period as controls.The relationships among the expression of IR,IR-A isoforms and the clinicopathological parameters of EC tissues were analyzed by Spearman rank correlation analysis.An eukaryotic IR-A expression plasmid was constructed and transfected into RL95-2 cells[RL95-2(IR-A)]for overexpression of IR-A in RL95-2 cells,in which the expression of IR-A originally was low.Non radioactive cell proliferation assay—MTS was used to determine the proliferation curves in the four EC cells listed above and in RL95-2 or RL95-2(IR-A).Results(1)There were two isoforms of IR-A and IR-B coexpressed were detected in EC cells and EC tissues.Among four kinds of EC cell lines,the expression level of IR mRNA in RL95-2 cells was the highest,followed by Ishikawa,KLE and HEC-1-A cells,in which the relative IR mRNA expression levels were(26.54 ± 1.82)× 10-4,(15.44 ± 3.29)× 10-4,(10.14 ±0.10)× 10-4 and(2.63 ±0.23)× 10-4,respectively(P ＜0.01).The expression level of IR-A mRNA was the highest in Ishikawa cells,followed by KLE,RL95-2 and HEC-1-A cells,with the relative expression levels were(12.07 ±3.31)× 10-4,(4.68 ±0.63)× 10-4,(3.03 ±0.22)× 10 4 and(1.46 ±0.03)×10-4,respectively(P ＜0.01).The relative expression level of IR and IR-A mRNA were 0.017 ±0.013 and 0.011 ±0.010 in the EC tissues,respectively,compared with 0.015 ± 0.014 and 0.010 ± 0.012 in the controls,in which there were no significant differences in the expression level of IR or IR-A mRNA between EC tissues and the control(P =0.662,P =0.780).The expression of IR and IR-A in EC tissues had no significant relevance with International Federation of Gynecology and Obstetrics(FIGO)stage,cell differentiation,depth of myometrial invasion,invasion of lymph-vascular space,lymph nodes metastasis,the expression status of estrogen receptor,human progesterone receptor,and PTEN gene(all P ＞ 0.05).The expression of IR-A mRNA in EC patients with type 2 diabetes mellitus(DM)was significantly higher than that in patients without type 2 DM(P =0.031),while there were no statistical correlation between the expression of IR mRNA and type 2 DM(P =0.438).(2)The proliferation rates of the four kinds of EC cells was positively related with the IR-A expression ratio,with the most growth potential in Ishikawa,followed by HEC-1-A,KLE and RL95-2 cells.The overexpression of IR-A in RL95-2(IR-A)cells showed a significant proliferation-promoting effect than that in control RL95-2 cells(P ＜ 0.01).Conclusion There are two isoforms of IR-A and IR-B co-expressed in EC.The overexpression of IR-A may promote the proliferation of EC cells.

Objective To study the role and mechanism of microRNA-16(miR-16)in the proliferation,invasion and apoptosis of ovarian epithelial carcinoma cells in vitro.Methods The SKOV-3 cells were transfected with miR-16 mimics or negative control RNA(NC)by lipofectamine 2000.The expression of miR-16 was detected by real-time reverse transcription(RT)-PCR in SKOV-3 cells,and western blot was used to detect the expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2(MMP-2)and bcl-2 protein.Methyl thiazolyl tetrazolium(MTT),5-ethynyl-2'-deoxyuridine(EdU)and transwell assay were used to determine the proliferation and invasion abilities.And the rate of apoptotic cell was detected by flow cytometry method.Results(l)The expression level of miR16 in the transfection cells group was significantly higher than that in NC group(125.93 ± 15.30 versus 0.78 ± 0.16,P ＜ 0.01).(2)The rclative expression level of VEGF protein in transfection cells,NC and blank control group was 0.58 ± 0.05,1.22 ± 0.03,1.20 ± 0.03,MMP-2 protein was 0.63 ± 0.03,1.16 ±0.03,1.21 ± 0.03,and bel-2 protein 0.52 ± 0.03,1.19 ± 0.05,1.28 ± 0.06,respectively.The level of VEGF,MMP-2 and bcl-2 protein in the transfection group were lower than those in other control groups,and there were significantly differences among them(all P ＜0.01).(3)After transfected 4 days,the inhibition rate of cell proliferation in the transfection group was dramatically higher than that in NC group[(37.2 ±6.2)％ versus(3.6 ± 3.2)％,P =0.001].(4)The percentage rate of proliferative cells in the transfection,NC and blank control group was(12.3 ± 0.8)％,(23.4 ± 1.8)％,(31.1 ± 4.9)％.And it was lower in the transfection group(P ＜ 0.05).(5)Decreased cells via the transwell member in the transfection group(6 ± 3)were detected as compared with NC group(40 ± 9)and blank control group (48 ± 8,P ＜ 0.01).(6)Twenty-four hours after cultured in serum starvation and hypoxia,the rate of the viable and late apoptotic cells in the transfection group were significantly higher than those in NC group and blank control group[the rate of viable apoptotic cell was(16.9 ± 2.1)％,(10.3 ± 1.7)％ and(9.0 ±0.8)％ respectivcly,P＜0.01;the rate of late apoptotic cell was(13.4±3.3)％,(3.2 ±1.8)％ and (0.7 ±0.6)％ respectively,P ＜ 0.01].After cultured 48 hours,total apoptotic cells in the transfection group was significantly more than those in other groups(P ＜ 0.01).Conclusion miR-16 might inhibit the proliferation,invasion of ovarian epithelial carcinoma cells and enhance their sensitivity to apoptotic stimuli via downregulation of the expression of VEGF,MMP-2 and bcl-2 protein.