Vascular endothelial growth factor expression and angiogenesis in human ovarian tissue after different cryopreservations / 中国组织工程研究

BACKGROUND: The cryopreservation of human ovarian tissue has become an attractive method to preserve female fertility. Human ovarian tissue experiences neovasculadzation after transplantation to recover blood supply, cryopreservation and resuscitation technique is a key for the neovascularization of human ovarian tissue following transplantation. OBJECTIVE: To investigate vascular endothelial growth factor (VEGF) expression and microvessel density in human ovadan tissue following novel needle immersed vitrification (NIV) and slow-freezing, to explore the influence of two cryopreservation methods play in the neovascularization of human ovarian tissue after transplantation. METHODS: Eight normal human ovarian tissues from patients with carcinoma of endometrium were cut into 12 fragments in the size of 1.5 mm × 1.5 mm × 1.0 mm, then randomly assigned to 3 groups: fresh control group, NIV group and slow-freezing group. In the NIV group, pieces of ovarian tissue stdps were dehydrated in an equilibration solution consisting of 7.5% ethylene glycol and 7.5% dimethyl sulfoxide in TCM-199 supplement with 20% fetal bovine serum and a vitrification solution consisting of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 mol/Lsucrose, then were plunged in liquid nitrogen directly and sealed in liquid nitrogen-filled cryovials. For thawing, the needles holding ovadan tissues were serially transferred into 1.0, 0.5, 0.25 mol/L sucrose solution and TCM-199 supplemented with 20% fetal bovine serum. In the slow-freezing group, pieces of human ovadan cortex fragments were placed in a 1.8-mL cryovial containing 1 mL of TCM-199 medium supplemented with 0.1 mol/L sucrose, 20% fetal bovine serum and 1.5 mol/L dimethyl sulfoxide, the cryovials were placed in the programmable freezer and cryopreserved by preset slow-cooling protocol. For thawing, the ovarian tissue stdps were washed in a stepwise manner: 1.0 mol/L dimethyl sulfoxida + 0.1 mol/L sucrose, 0.5 mol/L dimethyl sulfoxide + 0.1 mol/L sucrose, 0.25 mol/L dimethyl sulfoxJde + 0.1 mol/L sucrose and 0.1 mol/L sucrose. The frozen-thawed and fresh controlled human ovarian tissues were cultured in vitro. The expression of VEGF and CD34, as well as microvessel density, was analyzed by immunohistochemistry. RESULTS AND CONCLUSION: There was patchy and mild expression of VEGF in the stromal cells of all the three groups before and after culture. The expression of VEGF increased and reached peak value after culture for 2 days, began to decrease after culture for 4 days and further attenuated after culture for 6 days in all the three groups. Compared with slow-freezing group, the expression of VEGF in NIV group was closer to that in fresh control group. Microvessel density of all the three groups increased and reached peak value after culture for 2 days, and the microvessel density of fresh control group and NIV group was significantly higher than that of slow-freezing group (P < 0.05). The microvessel density of slow-freezing group after culture for 4 days and that of all the three groups after culture for 6 days significantly decreased compared with after culture for 2 days (P <0.05). NIV is superior to slow-freezing to preserve stromal cells and extracellular matrix of human ovarian tissue, and plays less influence in VEGF expression and angiogenesis in human ovarian tissue.

A multicenter prospective randomized open comparative study on the treatment of ovulatory menorrhagia with tranexamic acid and norethisterone in China / 中华妇产科杂志

Objective To compare the efficacy and safety of tranexamic acid(TA)and norethisterone(NET)for the treatment of patients with ovulatory menorrhagia in China. Methods Onehundred and thirty one patients with proven ovulatory menorrhagia from gynecologic clinics of 5 teaching hospitals located in 4 different cities in China were enrolled during Jul 2004 to Dec 2006.Ameng them 128 completed the study.Patients were randomly divided into two therapeutic regimen groups:TA 1g thrice daily during menstrual cycle days(D)1-5,69 cases;or NET 5 mg twice daily on D19-26.59 cases.The drugs were administered for 2 consecutive cycles,then withdrawn and patients were followed-up for 1 more cycle.Data on menstrual blood loss [ estimated by pictorial blood assessment chart(PBAC)],length of menstrual periods,quality of life(QOL)evaluated by a 6 item health-related questionnaire were collectedbefore,during each cycle and were compared.Results Both treatments led to significant decreases of mean PBAC scores and shorter duration of menstrual periods,and improved the QOL ranking during the twotreatment cycles.The mean percentages of PBAC decrements in the TA first and second cycles were significantly greater than those in the NET corresponding cycles(35%VS 17%,P=0.004;4J4%VS 34%,P=0.04 respectively).The success rate of TA second cycle was higher than that of the NET second cycle (41%VS 24%,P=0.04).Improvement of QOL ranking in the TA first cycle was also significantly better than those in the NET first cycle ( P=0.03).The percentage of patients with at least 1 adverse event in TA group(19%)was significantly lower than that in NET group(35%,P=0.04).Patients'willingness tocontinue the treatment in the TA second and follow-up cycles(94%,79%respectively)were significantly higher than those in the corresponding cycles of NET groups(79%,59%respectively;P=0.01,P=0.02).Conclusion The regimen of TA 3 g daily during menstrual days 1-5 is a more effective and tolerable treatment than luteal phase norethisterone for patients with ovulatory menorrhagia.

Estradiol Profiles During the Luteal Phase in Patients Undergoing IVF-ET Cycles / 现代生物医学进展

Objective: To investigate the predictive value of serum E2 in early diagnosis of pregnancy through IVF-ET(in vitro fertilization-embryo transfer). Methods: Sixty-two patients (75 cycles involved) undergoing IVF-ET cycles, whose infertility was attributed to uterine tube or male factors, were enrolled. Luteal phase serum E2 was detected every other day after extraction of oocytes with micro-particle enzyme immunoassay. Results: The level of serum E2 declined progressively after extraction of oocytes in both pregnant and non-pregnant cycles, while it showed no significant difference on 2, 4, 6, 8 days after extraction of oocytes. In pregnant cycles following IVF-ET, serum E2 achieved the nadir on day 10 and then increased gradually. The differences of serum E2 levels in pregnant and non-pregnant cycles were detectable from day 10 after extraction of oocytes (816.4+ 537.6 vs. 189.5±69.3 pg/ml) (P＜0.05). In non-pregnant cycles, E2 on day 10 was significantly lower than that on day 8 ( 189.5+ 69.3 vs. 989.2+581.5 pg/ml) (P＜0.05). Conclusions: We concluded that the level of E2 on day 8 and 10 consecutively after extraction of oocytes may have predictive value in diagnosing early pregnancy. Ascension of serum E2 level of pregnant patients on day 10 forebodes the success of pregnancy, or else the failure of pregnancy.

Primary study on the method for separating mature sperms from human testicular tissue / 生物医学工程学杂志

This study was aimed to explore a simple and applicable method of separating mature sperms from human testicular tissue for intracytoplasmic sperm injection and embryo transfer. The suspension of human testicular tissue was cultured in 10% human serum albumin and human tubule fluid with different concentrations (0 u/ml; 50 u/ml; 100 u/ml; 150 u/ml; 200 u/ml) of hyaluronidase for 24 h, and then the Percoll gradient centrifugation was processed to separate the sperms; meanwhile the sperms were counted and graded according to their motility. The difference in quality and quantity among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treated groups contained large quantity and high quality of sperms as compared with the two contrast groups (P<0.01). The groups in the solution of 50 u/ml, 100 u/ml and 150 u/ml concentrations of hyaluronidase had almost the same amount of sperms that displayed higher motility as compared against the sperms in the group treated with 200 u/ml concentration of hyaluronidase (P<0.01). There was no difference between the two contrast groups (P>0.05), or among the groups treated with 50 u/ml, 100 u/ml, and 150 u/ml of hyaluronidase concentration (P>0.05). This method of adopting hyaluronidase with Percoll gradient centrifugation in the process for separating mature sperms from human testicular tissue is applicable. It can increase the quantity and quality of sperms separated from testicular tissue suspensions when adequate concentrations of hyaluronidase is used.

Advance in oriented differentiation of embryonic stem cell / 生物医学工程学杂志

Embryonic stem cells have been the focus in tissue engineering, developmental biology, drug development and gene research. In this paper, the mechanisms, advances and applications of oriented differentiation of embryonic stem cells are reviewed for the promotion of researches on embryonic stem cells in future.