BACKGROUND: Nowadays stem cells have been isolated successfully from the epithelial tissue of human and rodents such as skin, hair follicle, cornea, oral mucous, intestinal mucosa dental epithelial cells, and salivary glands by the Chinese or foreign researchers. But according to the vaginal mucosa stem cells (VMSCs), it is not known in the field of isolation and application.OBJECTIVE: To find out the suitable method of isolation, identification, and culture of human VMSCs.DESIGN: Observational basic study. SETTING: Department of Gynaecology and Obstetrics, the Third Affiliated Hospital of Guangzhou Medical College. MATERIALS: This study was performed in the Zhongshan Ophthalmology Center of Zhongshan Medical College from October 2004 to August 2005. Two-week gestation mice were provided by the Experimental Animal Center of Guangzhou University of Chinese Medicine. The sterile human vaginal mucosa was collected from five patients (25-38 years old) after the plastic operation of vagina from the 3rd Affiliated Hospital of Guangzhou Medical College. METHODS: Vaginal mucosa cells could be harvested from human vagina mucosa treated with trypsin-collagenase digesting. The VMSCs were selected by adhesion to type Ⅳ collagen for 20 minutes. The cell cycles of the VMSCs and primary human vaginal mucosa cells were analyzed by flow cytometry. VMSCs were cultured in vitro with 4 different media, including feeder layer cells plus epidermal total culture media, feeder layer cells plus DMEM/F12 (3:1), epidermal total culture media and DMEM/F12 (3:1). At the 12th day, their colony forming efficiency (CFE), CK19 and CK10 positive rates were compared. Then VMSCs were continuously cultured and passaged till they were apoptosis.MAIN OUTCOME MEASURES: ① Positive rate of CK19 and CK10 and G0/G1 proportion; ② Positive rate of CK19 and CK10, the colony forming efficiency (CFE) and the times be passaged when primary VMSCs were cultured in different media after 12 days. RESULTS: The cells at the G0-G1 phase cell cycle, CK19 and CK10 positive rates between the VMSCs and primary human vaginal mucosa cells were statistical different (P < 0.05). When cultured at 12th day, the VMSCs cultured with feeder layer and epidermal total culture media had the highest CFE and CK19 positive rate and lowest CK10 positive rate. They could be passaged over 15 times in vitro. CONCLUSION: It is an effective way that select VMSCs by adhesion to type Ⅳ collagen for 20 minutes. In this research, culturing VMSCs in vitro with feeder layer and epidermal total culture media was the best way to keep VMSC characteristics.

Objective To compare the efficacy of gradient and swim-up semen preparation techniques on pregnancy rates in couples undergoing intrauterine insemination (IUI) cycles with human menopausal gonadotropin (HMG) stimulation. Methods Five hundred and seventy one cycles were devided into the swim up or the gradient technique groups for sperm preperation. The clinical pregnancy rates per cycle were evaluated. Results The clinical pregnancy rates per cycle were significantly higher in the gradient group (17.8%) than that in the swim up group (11.4%)(P < 0.05). In the subgroup of unexplained subfertile couples, the gradient group also revealed significantly higher clinical pregnancy rates per cycle (33.7%) than that in the swim up group (20.3%)(P<0.05). In couples with mild male factor subfertility, the gradient group also revealed significantly higher clinical pregnancy rates per cycle (11.6%) than that in the swim up group (7.6%)(P < 0.05). Conclusion The gradient technique significantly improves clinical outcome in IUI cycles of unexplained subfertile and male subfertile couples.

Objective To explore the lowest effective dosage of mifepristone combined with misoprostol in terminating ultra-early pregnancy.Methods All the cases of ultra-early pregnancy classified by amenorrhea days,β-hCG and vaginal B-ultrasonic were randomly divided into two groups.One hundred cases in G1 group (minimized dosage) were orally administered 25 mg mifepristone once a day for 2 days and combined with 200 μg misoprostol 48 hours later,while 150 mg mifepristone combined with 600 μg misoprostol 48 hours later were given to 100 cases in G2 group (normal dosage).All cases were observed for 6 hours after taking misoprostol and returned for assessment three days later.Results None missing.Expulsion of conceptus:G1 and G2 group were 22 (22.0％,22/100) and 25 (25.0％,25/100;P ＞ 0.05).Failure rate:cases with incomplete abortion were 1 (1.0％,1/100) and 2 (2.0％,2/100) in G1 and G2 group,hospitalization for suspected ectopic pregnancies both was 1 (1.0％).Bleeding:bleeding cases during the administration of mifepristone in G1 and G2 group were 71 (71.0％,71/100) and 78 (78.0％,78/100; P＞0.05); the mcan bleeding time were (5.3 ± 1.4) days and (6.0± 1.5) days (P ＜0.01).Other side effects:in G1 group,majority showed light nausea (7.0％,7/100) and light abdominal pain (20.0％,20/100).Menses recovery:99 (99.0％,99/100) for G1 group and 98 (98.0％,98/100) for G2 group to recovery on scheduled time.Satisfactions:both were 99 (99.0％,99/100).Except mean bleeding days and side-effects,the differences above showed no significance (P ＞ 0.05).Conclusion It is safe and effective treatment with the lowest dosages of mifepristone and misoprostol to terminate ultra-early pregnancies.

Objective To explore the efficiency and the clinical application value of non-invasive prenatal genetic testing for fetal chromosomal aneuploidy.Methods A total of 1 865 pregnant women treated in Guangdong Women and Children Hospital from January 201 1 to January 2013 were selected.Inclusion criteria:advanced age,prenatal screening for high risk,and fetal abnormality indicated by color ultrasonography,agreeing with non-invasive prenatal genetic testing.After non-invasive prenatal genetic testing, the pregnant women with positive result underwent cell culture and chromosomal karyotyping.Following the situations after deliv-ery were designed as the final criteria for definite diagnosis of fetal chromosomal aneuploidy.Results A total of 1 865 pregnant women underwent non-invasive prenatal genetic testing,of which 21 pregnant women were found with positive result,including 14 pregnant women with trisomy 21,5 pregnant women with trisomy 18,2 pregnant women with trisomy 13.The results of chromo-somal karyotyping after amniocentesis or umbilical cord blood puncture were designed as golden standard.Among the women with trisomy 21,one woman refused the prenatal diagnosis,self induced labor and could not be confirmed karyotype.No false positive case was found among the women with trisomy 18 and 13.No missed diagnosis was found among the pregnant women with negative result during follow-up after delivery.Through statistical analysis of non-invasive prenatal fetal genetic testing,the sensitivity for the trisomy 21 was 100%,and the accuracy was 92.9%.The sensitivity and accuracy for the trisomy 18 and 13 were 100%.Conclu-sion Non-invasive prenatal genetic testing can improve the diagnostic efficacy before delivery,reduce the birth of ill infants,and it is a quick,safe,easy-accepted and reliable prenatal diagnostic method,which is worthy to be popularized and an inexorable trend of development in the future.

Objective To compare the parameters of bilateral tubal ligation by LESS and multi-incision laparoscopy and their prognosis of IVF-ET. Methods From Jun. 2016 to Dec. 2017,32 patients underwent LESS bilateral tubal ligation and 60 patients under-went multi-incision laparoscopic bilateral tubal ligation were enrolled. The demographic data ,operative parameters ,and postoperative prognosis of IVF- ET in the two groups were summarized and compared. Results The mean operating time in LESS group was prolonged and its operative hospital stay was significantly decreased compared to multi-incision laparoscopic group. No significant difference was observed in the intraoperative blood loss in two groups. Moreover,no significant differences were found in the AFC,AMH,Gn stimulation days,total Gn dosage,Ovum number,good quality embryo rate,embryo implantion rate,clinical pregnancy rate and early spontaneous abortion rate in two groups. Conclusions For experienced laparoscopic surgeons,LESS is a safe and feasible procedure with similar perioperative outcomes to multi-incision laparoscopic surgery.