One of the focuses in bone and cartilage tissue engineering research is to find suitable seed cells. Embryonic stem cells derived from the inner cell mass of blastocysts possess unique characteristics such as unlimit- ed capacity of proliferation and the potential to differentiate into diverse cell types (pluripotentiality), which make them a promising cell source for tissue engineering. In this paper, the progress in the study on embryonic stem cell differentiating directly into osteoblasts and chondrocytes is introduced. The prospects and difficulties of using em- bryonic stem cells as seed cells for tissue engineering are also discussed.

Objective To establish clonal human embryonic stem cell lines and investigate their biological characteristics. Methods Cells were derived from one inner cell mass of human blastocyst, multiplied for 20 passages, and then dissociated into single cell suspension by digestion with 0.5% trypsin. Single cell was picked up and plated into individual well of a 96-well plate containing feeder-layers directly under a dissection microscope. The outgrowth clones were passed by treatment with collagenase. Surface markers were detected by cytochemistry and histoimmunochemistry. Karyotypes were tested using standard G-banding techniques. The pluripotency was analyzed by inoculating cells into severe combined immunodeficient (SCID) mice. Results Two clonal human embryonic stem cell lines were established. Cells of these two lines possess the characteristics and differentiating potencies: normal 46 XX karyotypes; expressing a series of surface markers such as: alkaline phosphotase, stage-specific embryonic antigen (SSEA)-4, tumor recognition antigen (TRA)-1-60, TRA-1-81 etc; and forming teratomas comprising derivatives of three embryonic germ layers such as neural tissue, cartilage, squamous epithelium and columlar epithelium when injected into SCID mice. Conclusions The two single cell-cloned human embryonic stem (hES) cell lines were derived in our laboratory. The cells possess stable biological characteristics of undifferentiated hES cells.

Objective To investigate the association between renin-like activity (RA) and angiotensin Ⅱ (AⅡ) and the ovarian hyperstimulation syndrome (OHSS). Methods Blood samples were taken from 42 patients undergoing in vitro fertilization (IVF). According to the ovarian stimulating response, the patients were divided into 4 groups: group Ⅰ, low responders, 7 cases; group Ⅱ, moderate responders, 8 cases; group Ⅲa high responders without using albumin, 7 cases, group Ⅲb, high responders with albumin, 10 cases; group Ⅳ, severe and moderate OHSS, 10 cases. 5 patients for intrauterin insemination with natural cycle as control. Follicular fluid (FF) was collected at oocyte retrieval from 28 IVF patients (including 7 OHSS patients). Fluid from ascites and hydrothorax was obtained from 3 OHSS patients, and peritoneal fluid obtained from 5 infertility patients by laparoscopy and ascitic fluid obtained from 6 ovarian carcinoma patients served as control. RA and AⅡ levels were measured by radioimmunoassay. Results plasma RA and AⅡ levels at mid-luteal phase from OHSS patients [(19.9?19.0)?g?L -1 ?h -1 and (397.0?378.2) ng/L] were significantly higher than those from the other patients ( P

Objective To detect the expression of the vascular endothelial growth factor (VEGF) and its receptors, the fms like tyrosine (flt 1), kinase insert domain containing receptor (KDR) in normal human endometrium during menstrual cycle and investigate the functions of VEGF and its receptors for development and differentiation of human endometrium Methods Immunohistochemistry and in situ hybridization techniques were used to measure the level of VEGF, flt 1 and KDR protein and mRNA in normal endometrium from 50 women Few samples of endometrium respectively in the proliferative and secretory phase were homogenized for Western Blot analysis of VEGF subtypes The blood vessel density was estimated by counting staining with a endothelial marker Ⅷ factor Results The VEGF and its receptors mainly expressed in endometrial endothelial cells and gland epithelium The level of VEGF, protein and mRNA, as well as flt 1 in mid secretory and menstrual phase were highest ( P 0 05) Western Blot analysis showed stained VEGF bands in 34 000 (VEGF 121 ), 46 000 (VEGF 165 ), 54 000 (VEGF 189 ), 68 000 (VEGF 206 ), predominantly in 34 000 and 46 000 Conclusions VEGF, flt 1 and KDR, including both protein and mRNA, showed a pronounced menstrual cycle dependent expression in cycling endometrium VEGF and its receptors expressing higher in secretory phase and menstruation probably be involved in embryonic implantation and endometrial shedding

Objective To investigate the incidence of follicular stimulating hormone receptor (FSHR) gene C566T mutation in Chinese women with premature ovarian failure (POF) and to explore the etiologies of POF. Methods This case-control study was carried out between 73 Chinese women with idiopathic POF (POF group) and 35 controls (control group), including 25 normal females with a regular menstrual history and 10 normal post-menopause women. DNA was extracted from the peripheral blood of patients and controls. The exon 7 of FSHR gene was amplified by PCR. PCR products were subsequently digested by the enzyme BsmI and then subjected to electrophoresis on agarose gels and stained with ethidium bromide to determine the C566T mutation. DNA samples of random sampling were further analysed by sequencing the PCR products to confirm the mutation. Results BsmI digestion resulting in two fragments of 51 and 27 base pairs was noted for all 73 POF patients and 35 controls. PCR sequencing confirmed that the 566 allele of FSHR gene is C, demonstrating normal FSHR allele. Conclusions No FSHR gene C566T mutation is present in POF patients and controls. FSHR C566T mutation may be rare in Chinese women with POF.

Objective To study expression of cystic fibrosis transmembrane conductance regulator (CFTR) in human endometrium. Methods The expression of CFTR mRNA and protein from 50 samples of normal cyclic human endometrium was examined by in situ hybridization, immunohistochemistry and Western blotting respectively. Results CFTR mRNA and protein expressions were only detected in the endometrial glandular cells with cyclic changes. CFTR mRNA could be detected from the midproliferative phase with the highest level found in the late proliferative phase, significantly higher than those of late-secretory phase endometrium ( P

0.05).The pregnancy rate was 41.5%(17/41) in the(Non-cystectomy) Group,33.3%(23/69) in the Laparoscopic Group,and 25.5%(14/55) in the Open Group,respectively,without significant differences(?~2=2.754,P=0.252).A total of 86 patients(90 cycles) were given a unilateral cystectomy,including 37 cycles in the Open Group and 53 cycles in the Laparoscopic Group.In the 37 cycles of the Open Group,the number of dominant follicles was less in affected ovary(4.41?4.02) than in contralateral one(6.14?4.37)(t=-2.364,P=0.024),whereas in the 53 cycles of the Laparoscopic Group,the number of dominant follicles was significantly less in affected ovary(3.33?3.50) than in contralateral ovary(6.40?3.61)(t=-5.358,P=0.000).Conclusions Both laparoscopic and open cystectomy of ovarian endometrioma may cause damage to ovarian response of COH.

Objective To investigate the effects of mifepristone on apoptosis and expression of relative genes in early pregnant chorionic villi and decidua Methods The specimen of early pergnant chorionic villi and decidua obtained from 10 cases of requesting temination of pregnancy by curettage, 20 cases of mifepriston contragestation The paraffin sections were used to determine apoptotic cells by TdT mediated dUTP biotin nick end labeling method, to identify bcl 2, bax, fas, fasL and proliferating cell nuclear antigen (PCNA) by immunohistochemistry, to demonstrate fas and fasL mRNA by in situ hybridization Results In normal early pregnant specimens, apoptotic cells were mainly observed in syncytiotrophoblast, but not in cytotrophoblast cells, occationally seen in decidua cells The antigen of bax, fas, fasL were present in syncytiotrophoblast cells and decidua with lower amount While bcl 2 antigen staining was strong in cytotrophoblastic cells and in decidua PCNA protein was present in cytotrophoblastic and decidual cells only In the specimens treated with mifepristone, apoptotic cells were increased in syncytiotrophoblastic cells of villi and visualized in decidua cells The expression of fas, fasL and bax was also higher than that of nomal Conclusions Mifepristone increased apoptosis in syncytiotrophoblastic and decidua cells, but had no effect on the expression of bcl 2 and PCNA

Objective To study the expression of matrix metalloproteinase(MMP)-2,9 and tissue inhibitor of metalloproteinase(TIMP)-1,2 protein in human endometrial carcinoma tissue and its relation to the invasion and metastasis of endometrial carcinoma. Methods Immunocytochemistry and zymography techniques were used to measure the MMP-2,MMP-9,TIMP-1,TIMP-2 protein levels and activities in endometrial carcinoma tissue of 37 patients and control group composed of 7 normal postmenstrual endometrial samples. Results The MMP-2,MMP-9,TIMP-1 and TIMP-2 proteins mainly expressed in endometrial carcinoma cells, glandular cells and endothelial cells. The strongly positive expression proportions of MMP-2,9 and TIMP-1 proteins in grade Ⅲ carcinoma cells were respectively 73%, 20% and 67%, which were higher than those in gradeⅡ (13%, 0, 27%) and gradeⅠ (0) ones ( P

Objective To compare the clinical outcomes of gonadotropin-releasing hormone (GnRH) antagonist (GnRH-ant) fixed protocol with GnRH agonist (GnRH-a) long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer (IVF-ET) cycle,and to explore the feasibility and advantage of GnRH antagonist protocol performed in normal responders.MethodsFrom January 2011 to June 2011,771 infertile women with normal ovarian reserve function underwent their first IVF or intracytoplasmic sperm injection (ICSI) cycles in Peking University Third Hospital,which were divided into 245 cycles in GnRH-ant fixed protocol group ( GnRH-ant group) and 526 cycles in GnRH-a long protocol group ( GnRH-a group).The data of general demographic,treatment and clinical outcome were compared between two groups.ResultsAge,infertile duration,body mass index (BMI),baseline serum follicle-stimulating hormone (FSH) and estradiol levels between two groups did not reached statistical difference (P ＞ 0.05 ).The level of estradiol was (12 289 ± 6856) pmol/L in GnRH-ant group and (14934±8007)pmol/L in GnRH-a group at day of hCG injection.The mean length of stimulation was ( 10.3 ± 1.2) days in GnRH-ant group and ( 12.8 ± 1.6) days in GnRH-a group.The dose of gonadotropin was (2013 ± 607 ) U in GnRH-ant group and (2646 ± 913 ) U in GnRH-a group.The number of ovum was 15 ± 7 in GnRH-ant group and 17 ± 8 in GnRh-a group.Those clinical parameter all reached statistical difference (P ＜0.05 ).The number of embryo was 7 ±4 in GnRH-ant group and 8 ± 5 in GnRH-a group,the rate of clinical pregnancy was 40.9％ (94/230) in GnRH-ant group and 45.6％ (216/474)in GnRH-a group,the rate of implantation was 26.1％ (128/490)in GnRH-ant group and 30.9％ (307/994) in GnRH-a group,the rate of continuing pregnancy was 38.7％ ( 89/230 ) in GnRH-ant group and 42.6％ (202/474) in GnRH-a group,those parameter did not reach statistical difference (P ＞ 0.05).The rate of moderate or severe ovarian hyperstimulation syndrome was 2.4％ ( 6/245 ) in GnRH-ant group and 4.2％ (22/526) in GnRH-a group,which did not show significant difference ( P ＞ 0.05 ).ConclusionIn the first IVF or ICSI cycle of the patients with normal ovarian reserve function,the fixed GnRH-ant protocol could get the same satisfied clinical outcome,and it is more economic,convenient and safer compared with low dose depot GnRH-a long protocol.