BACKGROUND:Numerous studies have confirmed that neovascularization plays an important role in the growth, invasion and metastasis of tumors. OBJECTIVE:To investigate the features of CD133+ ovarian cancer stem-like cels differentiating into vascular endothelial cels. METHODS:CD133+ ovarian cancer stem-like cels were successfuly harvested from A2780 ovarian cancer cel lines using serum-free culture method, and incubatedin vitro onto 96-wel plates with or without Matrigel. Then, we observed the capacity of CD133+ ovarian cancer stem-like cels and human umbilical vein endothelial cels to form tube-like structures at different time points. Through xenograft experiments, the role of CD133+ ovarian cancer stem-like cels in the angiogenesis of ovarian cancer was observed using immunofluorescence staining. RESULTS AND CONCLUSION:CD133+ovarian cancer stem-like cels and human umbilical vein endothelial cels cultured with no Matrigel had no corresponding lumen formation, and could not express CD31. But those cultured with Matrigel had lumen formation and expressed CD31 significantly. After tumor formation, human-derived CD31 expression was observed in the tumors. These findings indicate that CD133+ ovarian cancer stem-like cels can differentiate into vascular endothelial cels, and be involved in tumor revascularization.

BACKGROUND:Placental mesenchymal stem cel s with rich sources are similar to bone marrow mesenchymal stem cel s in terms of morphology, surface markers and differentiation potential, which are one of ideal mesenchymal stem cel s in human body. However, there are few studies addressing the ultrastructure and phagocytotic function of human placental mesenchymal stem cel s and its physiological role in the the placenta has been little explored. OBJECTIVE:To investigate the ultrastrcture and phagocytotic function of placental mesenchymal stem cel s. METHODS:Placental mesenchymal stem cel s obtained from five placentae of normal pregnancy were cultured in vitro and observed for ultrastructure under transmission electron microscope. The fluorescent beads were added in the supernatant for 3 hours, and then the phagocytosis of placental mesenchymal stem cel s was evaluated by flow cytometry. RESULTS AND CONCLUSION:Under the transmission electron microscope, placental mesenchymal stem cel s had large nuclei with prominent nucleoli. In the cytoplasm, a plenty of rough endoplasmic reticula was seen, dilated or stacked. The cytoplasm was also rich in Golgi apparatus and lysosomes. The cel surfaces were covered by microvil i. The intercel ular junctions could be seen occasional y. A part of cel s from these five samples could phagocytose fluorescence beads, which ranged from 49.6%to 18.4%. The ultrastructural characteristics of placental mesenchymal stem cel s suggested these cel s were active to synthesize and secrete proteins and had phagocytotic function, indicating placental mesenchymal stem cel s may play a role in keeping the balance of micro-environments and clean the foreign substances in the placenta.

OBJECTIVETo explore the relationship between HSD11B2 polymorphisms and fetal growth during normal pregnancy.

METHODSThe HSD11B2 promoter/G-209A, G-194C, G-151A and G-126A genotypes were examined in 33 samples from Chinese Han subjects by gene sequencing. HSD11B2 (CA)n microsatellite polymorphism in the first intron was detected in blood samples from 187 maternal and newborn pairs by PCR-capillary electrophoresis.

RESULTSAll the HSD11B2 promoter/G-209A, G-194C, G-151A and G-126A genotypes were wild-type GG. The offspring birth weight and any ultrasound parameters describing late gestational fetal body shape were not significantly different between maternal or fetal SS, SL and LL groups or combined SS+SL and LL groups. When considering the relevant confounding factors (gestational age at delivery, newborn's gender, maternal body mass index before pregnancy, maternal weight at delivery and maternal age), the offspring birth weight and late pregnancy ultrasound parameters were still not associated with the maternal or fetal HSD11B2 (CA) n microsatellite polymorphisms.

CONCLUSIONSFetal and maternal HSD11B2 polymorphism is not related to fetal growth during normal pregnancy.

11-beta-Hydroxysteroid Dehydrogenase Type 2 ; genetics ; Birth Weight ; Body Mass Index ; Female ; Fetal Development ; genetics ; Genotype ; Gestational Age ; Humans ; Infant, Newborn ; Polymorphism, Genetic ; Pregnancy ; Promoter Regions, Genetic

BACKGROUND:Cytotrophoblasts in placental cell components plays an important role in fetal immunological tolerance.Placental mesenchymal stem cells(pMSCs)have potential of multiple differentiation and inhibition of lymphocyte proliferation.However,conventional methods cannot acquire a large amount of purified human cytotrophoblasts or pMSCs.OBJECTIVE:To establish a method to obtain large placenta tissue,and harvest plenty of cytotrophoblasts and pMSCs with high purity and activity.METHODS:Human placenta tissues were dissected,minced,and dissociated in trypsin and DNAse I.The dissociation was performed in three stages of incubation at 180 r/min for 20 minutes at 37 ℃ The digesting suspension was filtered using a 200 mesh strainer before separated by Percoll gradients.The cytotrophoblast cells and pMSCs fractions were collected respectively.Fibroblasts of cytotrophoblast cells fraction were removed by differential adhesion.The pMSCs were seeds on 75-cm2 flask directly for culture.The dissociation of placenta tissue was observed.The number of harvesting cytotrophoblasts was quantified and Cytokeratin 7 expression was tested.The pMSCs primary culture time,cell passage,induced osteoblast differentiation were observed.The cell surface makers were also detected.RESULTS AND CONCLUSION:After digesting in trypsin and DNAse I,there was only little residue left.(5.48±1,98)×10~8 cytotrophoblasts were obtained after differential adhesion.(90±4.36)% of these cells were positive for Cytokeratin 7.At 19-21 days after pMSCs reached approximately 90% confluency,the cell number was(1.96±0.24)×10~6.The subcultre cells could be passaged again in 4 or 5 days.Flow cytometric analysis of pMSCs showed that the cells expressed CD29,CD44 and HLA-ABC intensively and were negative for CD34,CD45,CD14 and HLA-DR.pMSCs differentiated into osteoblast-like cells after induction,which stained bright salmon pink by Alizarin Red.Dissociating the placenta tissue in trypsin and DNAse I in combination with discontinuous Percoll gradient separation obtained a large number of cytotrophoblasts and pMSCs recovered from placenta tissue,with high purity and activity.

Objective To compare the effects of resuscitation at pure oxygen environment(100% oxygen) or room air environment (21% oxygen) on the uhrastructure of cerebral cortical neurons in hypoxic neonatal rat.Methods Twenty Sprague-Dawley neonatal rats aged 7 days were divided into the pure oxygen environment (POE) and room air environment groups (RAE) after hypoxia,and each group was divided again into 24 h and 72 h subgroups.The pups were placed in an incubator containing 8% O_2 to be at acute hypoxia continued for 2.5 h.The pups were reoxygenated immediately after the above mentioned hypoxia.The pups of POE group were then placed in an incubator containing 100% oxygen and the flow time was 30 rain.The pups of RAE group were reoxygenated with room air in an incubator for 30 rain.According to the planned time points the pups were sacrificed and the brain was removed at 24 h and 72 h after treatment,respectively.The tissue of frontoparietal cortex of the fight cerebral hemisphere was prepared for transmission electron microscopic examination.Results In each reoxygenation group,edema was found in the neurons,neuropil and intercellular space.In the RAE 24 h group,the nuclear membrane in neurons was unclear,the amount of cell organelles was reduced,the mitochondria were swollen with damages of cristae.The rough endoplasmic reticulum was dilatated or vaeuolized and with reduction of ribosomes.The Golgi complex was vacuolized.The number of lysosomes was obviously increased.In the RAE 72 h groups,the changes were similar to those of the former group,but apoptotic-like nuclei and necrotic neurons were more frequently seen. The cellular damages of POE 24 h group were milder than those of RAE 24 h group. The mitochondria and rough endoplasmie reticulum were more abundant and showed less pathological changes in the POE 72 h group as compared with those of RAE 72 h group. Conclusion The rats of POE reoxygenation display milder ultrastructural damages, less apoptosis, necrosis and edema in cerebral neurons than those in rats after RAE reoxygenation.The protective effect of POE resuscitation on cerebral damage of hypoxic neonate rats is superior to that after RAE resuscitation. This hypoxic neonatal rat model may serve as a suitable animal model for research on cerebral cortex neurons caused by hypoxia.

Objective To study the expression of Survivin protein and its relation with the expression of Bcl-2, Bax in the tropho- blasts of human normal placenta. Methods The normal placental tissues (8 -9week ,18-23 week and 37-40week) were fixed, embed- ded, sectioned, and Survivin, Bcl-2, and Bax in the trophoblasts were detected with immunnohistochemistry. Result As the gestational age advanced , the staining intensity of Survivin in the trophoblasts was significantly decreased from the first trimester group to the second trimes- ter group to the term group (PU value: 11.74±0.8,9.95±0.43,8. 83 ~ O. 67, respectively, P <0.01 ), while the staining intensity of Bcl-2 and Bax in trophoblast was significantly increased (PU value of Bcl-2 : 4.33±0.60, 5.00±0.75,6.87±0.45, respectively, P<0.01 and PU value of Bax: 9.82±1.12,16.00±1.05,27.48±2.10, respectively, P <0.01 ). Expression of Survivin in trophoblasts has no rela- tionship with the expression of Bcl-2 and Bax (P>0.05 ). Conclusion Survivin may take part in the development of human normal pla- centa through the way of suppressing the apoptesis in trophoblasts. Expression of Survivin in trophoblasts has no relationship with the expres- sion of Bcl-2 and Bax, which indicate that they regulate apoptesis of trophoblasts via different biological pathways.

Objective To develope both qualitative and quantitative analytic method of the four urinary markers,i.e.lactate,uracil,orotate and hippurate,from ornithine transcarbamylase deficiency(OTCD) by Gas Chromatography-Mass Spectrometry(GC-MS).Methods Urea in urine samples was decomposed with urease,and heptadecanoiate was added as internal standard,then protein was denatured with ethanol and removed by centrifugation.After evaporation,the residue was derivatized trimethylsilylly by BSTFA/TMCS,and analyzed by GC-MS.ResultsThe markers can be separated in total ion current profiles,with indentifications confirmed by mass spectra.The significantly elevated levels of lactate,uracil and orotate in urine from OTCD patient were droped to normal or subnormal levels,together with large amount of hippurate excretion in the urine,after clinical therapeutic measures,including introduction of benzoic acid,were performed.Conclusion GC-MS analysis of the urinary markers is a valuable tool for the diagnosis and evaluation of the therapeutic outcome of OTCD.

AIM： To investigate the tumorigenicity of lung cancer by learning the accumulation of p53 and the exposure of cytokeratin 18 neo-epitope(CK-18neo) related to the clinicopathological parameters in non-small cell lung cancer(NSCLC). METHODS: To detect the monoclonal antibodies of p53 and M30 CytoDEATH(specific antibody for CK-18neo) in 62 cases of NSCLC (included 29 cases of squamous cell carcinoma and 33 cases of adenocarcinoma) and 10 cases of control group by adopting immunohistochemistry assay (LSAB). Moreover, the immunoreactivity of p53 was quantitatively evaluated with positive unit (PU). RESULTS: (1) p53 immunoreactivity was positive in 15 of 29 squamous cell carcinoma (51.72%), 15 of 33 adenocarcinoma (45.45%), 30 of 62 NSCLC (48.39%). In 10 control cases was negative. There were significant differences between these groups (P＜0.01). (2)In 62 cases of NSCLC, AI% of M30 is 1.10%, and in 29 cases of squamous cell carcinoma is 0.95%, and in 33 cases of adenocarcinoma is 1.24%. In 10 control cases, the AI% is 1.06%. There is not significant difference among these groups . (3) According to the results of Pearson's correlation analysis, we found positive linear correlation between the immunoreactivity of p53（-/+）, p53（5 degrees）and p53（PU）(P＜0.01). CONCLUSION: Our results suggested that the pathogenesis of NSCLC might be related to the mutation of gene p53 and cell excessive proliferation and insufficient apoptosis.

AIM and METHODS:The purpose of the study was to characterize the time-related effect of Danazol therapy on endometriosis explant using the rat model. Endometriosis was induced in mature female rats. One group of treated animals as well as controls were sacrificed at 2,4,6 and 8 weeks after treatment at which time the explant was evaluated. RESULTS:Explant volume was significantly reduced in all treatment groups, and the effect was more significant in animals treated for 4 weeks than those treated for only 2 weeks. CONCLUSION:Danazol treatment can cause gradual regression of endometrial explant in a time-related manner.

Objective To investigate the effects of mifepristone on the ultrastructure of Hofbauer cells in human early pregnant placenta. Methods Twenty 6-9 week pregnant women with indications of pregnancy termination were recruited and randomlied to mifepristone (n=10) and D & C group (n=10).Villi were collected and studied with transmission electron microscope. Results In comparison with the control group,the ultrastructure of Hofbauer cells of mifepristone group showed the following changes: the cells were markedly edematous. The number of cytoplasmic processes of Hofbauer cells deceased obviously. In the cytoplasm of Hofbauer cells,the size of vacuoles enlarged and of mitochondrias minimized.Rough endoplasmic reticulum and Golgi complex were under-developed.Lysosomes were rare.The nuclei enlarged and showed irregular shape. Conclusions Mifepristone may change the phagocytosis'water and electrolyte transportation and immunological function of Hofbauer cells by influencing the ultrastructure of the Hofbauer cells.Therefore it can influence the development of pregnancy.