Study on the Estrous Cycle Regularity of Cryopreserved Rat Ovarian Tissues after Heterotopic Transplantation

Objective: To evaluate the re-initiation of ovarian function in cryopreserved ovarian grafts by means of vaginal smear of transplant rats. Methods: A total of 40 SPF-SD female rats (5 - 6 week-old) were randomly divided into three groups (blank control, castration control and transplant group). Ovaries were removed by surgical procedure then after cryopreservation and thawing procedures the ovarian tissues pushed inside the back muscles gap in transplant group. On the first PO day, vaginal smear collection was daily initiated. After 30 days, the PO day when the estrous cycle was re-initiated was considered for analysis as well as the estrous days and the number of estrous cycles. Results: Normal control group had a regular estrous cycle, while the transplant group had an estrous cycle disorder within first 2 weeks and later 2 weeks after the transplantation while the duration of diestrus cycle lasted longer. At the same time, the castration group had lost the normal estrous cycle, keeping continue in the stage of diestrus. Conclusion: In transplanted animals the re-initiated ovarian function can be predicted with alteration between estrous and diestrus phases with predominant estrous irregularity. Moreover, short autotransplanted graft duration needs time to perfuse by new blood vessels and hormone secretions, so could not directly affect its target organs to function properly.

Comparison of main lactobacillus species between healthy women and women with bacterial vaginosis

Background The normal microbial flora of the vagina plays an important role in preventing genital and urinary tract infections in women. Thus an accurate understanding of the composition and ecology of the ecosystem is important to understanding the etiology of these diseases. This study aimed to compare the characteristics of main lactobacillus species between healthy women and women with bacterial vaginosis （BV） by quantitative culture and PCR methods. Main lactobacillus species include L. crispatus , L. gasseri, L. jensenii and L. iners.Methods A total of 150 Women attending Gynecology Outpatient Clinic of Beijing Friendship Hospital, were diagnosed as having BV because three or more of the following criteria were met （standard of Amsel＇s composite criteria）： homogeneous discharge, elevated vaginal pH （pH 〉4.5）, production of amines, and presence of ＂clue＂ cells. Those with less than three of the criteria were considered as healthy. Simultaneously, smears were made of vaginal fluid and Gram stained, then were assessed qualitatively as normal （grade Ⅰ）, intermediate （grade Ⅱ）, or consistent with BV （grade Ⅲ）. Gardnerella vaginalis were identified by using Vitek 2 Compact and PCR methods. Lactobacillus species were identified by PCR methods. Gardnerella vaginalis and lactobacilli colony counts were determined by calculating the most number of colonies of each species in the appropriate plates （colonies between 10 and 300）, corrected by the dilution of the sample in the plates, and multiplied by 10 （to account for plating 100 μl）, in order to get colony forming units per milliliter of vaginal secretion.Results BV was diagnosed in 31% （46/150） patients using the composite criteria, the remainder being regarded as healthy. The majority of patients with BV had a smear assessed as grade Ⅲ （91%, 42/46） and minority of them had a smear assessed as grade Ⅱ（9%, 4/46）. The majority of healthy women had a smear assessed as grade Ⅰ （64%, 67/104）. Smears assessed as grade Ⅱwere found （36%, 37/104） among patients diagnosed as healthy, with less than three of the composite criteria. L. crispatus was cultured from 94% of healthy women and 83% of women with BV, with the former colonies count average value of 106 and the latter of 103. L. gasseri, L. iners, and L. jensenfi were cultured from 85%, 68% and 43% of healthy women; and 28%, 89% and 44% of BV women, respectively.Conclusions The quantities of four lactobacillus species except L. jensenii had a significant difference between healthy women and women with BV. Our results provide support for the negative association between L. iners and L. gasseri. Although L. crispatus were existent both in healthy and BV positive women＇s vagina, the numbers of L. crispatus were significantly different for the dominant number in healthy women. Smears of vaginal fluid and Gram stain play an important guiding role in bacteria culture.

Antifertility Activity of Hydroalcoholic Extract of Ocimum Basilicum Linn. Leaves on Female Wistar Rats

Objective To determine the effect of hydroalcoholic extract of Ocimum basilicum Linn. leaves on ovulation, implantation and maintenance of pregnancy in adult female cyclic Wistar rats. Methods Ocimum basilicum leaves extract （364 mg/kg and 624 mg/kg） was screened for its antiovulatory, antiimplantation and abortifacient activities. Vaginal smear of each rat was monitored daily during the 15-day treatment of the first experiment. Estimation of cholesterol in ovarian tissue and its histology was carried out to confirm its antiovulatory activity. In the second and third experiments, animals received treatment from 1st to 7th and 8th to 16th day of pregnancy, respectively. The number of implantations and litter size were determined through laparotomy on l Oth and 20th day of pregnancy, respectively in two tests. The treated groups were compared with pain control. Results A significant increase in duration of estrus cycle and diestrus phase was observed （P〈O.O01）. A significant decrease in ovarian weight and notable increase in ovarian tissue cholesterol level was also observed （P〈O. 001）. Weight of uterus in treated group was found to be decreased. Histological report showed presence of large corpora lutea in ovarian parenchyma. Neither antiimplantation nor abortifacient effect was observed in rats treated with both the doses of extract. Conclusion Hydroalcoholic extract of Ocimum basilicum interferes normal ovulation by disrupting the estrus cycle and prolonging diestrus phase thus, has a potential of being developed into a female contraceptive.