Objective:To explore the effects of Shenqi Compound Recipe on the expression of Cycloxygenase-2 (COX-2)mRNA in aorta in GK rats.There were five groups:GK group,model group,atorvastatin group,Shenqi Compound Recipe group and normal control group.During the experiment periods,each group was administrated correspondent substance respectively for 35 days.Serum concentrations of C reactive protein(CRP)were determined by ELISA.The mRNA expressions of COX-2 in aorta were detemined by reverse transcriptase PCR(RT-PCR).Results:Compared with model group,concentrations of CRP in serum and the mRNA expression of COX-2 all decreased in atorvastatin group and Shenqi Compound Recipe group(P

Objiective To evaluate the effect of pearventricular device closure of non-muscular ventricular defercts(VSDs) in infants.Methods From April 2007 to February 2008,39 patients with non-muscular VSDs were received off pump surgical treatment,perventricular device closure were for all the patients.There were 16 males and 23 females with mean age of(14.5±7.8) months(12 to 36 months and mean weight of(12.4±2.3)kg(8.5 to 18.0 kg).There were 34 perimembranous and 5 subateria VSD.The diameter of defects were 3.0 to 11 mm[mean(6.1±2.0)mm].Results Thirty-seven(94.9%)VSDs ere successfully closed,while two were repaired under cardiopulmonary bypass(CPB)instead of device closure because of the complication of moderat aortic reurgitation.The diameter of occluders were 4 to 12mm[mean(8.2±2.0)mm].The tricuspid regurgitations decareasded after operation in 3 patients with perimenmbranous VSDs,while the closure caused new mild or trace tricuspid regurgitations in 8 patients.Six patients with perimembranours VSDs acquired the incomplete right bundle branch blocks affter device closure.The length of hospital stay was 3 to 5 days[mean(3.4±0.4)]after operation and no petient had blood transfusion.Conclustion Perventricular device closure is probably an effective and safe treatment for non-muscular VSDs in unfants.

Objective To study the correlations of VDR FokⅠgene polymorphism with type 2 diabetes in postmenopausal women of Han nationality in south Sichuan. Methods 160 patients with type 2 diabetes (T2DM) and 190 healthy cases were enrolled in the study. The VDR FokⅠgene polymorphisms were detected using RFLP-PCR and DNA sequencing. Results The FF, Ff and ff genotype frequencies were 32.5%, 47.5%and 20% in the T2DM group and 15.8%, 53.7%, 30.5% in the control group, respectively (P < 0.05). The allele frequencies were 56.3%, 43.8% in the T2DM group and 42.6%, 57.4% in the control group, respectively (P < 0.05). The risk of T2DM in the FF genotype people was 2.568 times higher than Ff/ff genotype (adjusted OR = 2.568, 95%CI = 1.246 ~ 5.292, P < 0.05). The levels of 2 h PG and HbA1C in the FF genotype people were significantly higher than those of the Ff/ff genotype people (P<0.05). Conclusions There was an association between the VDR FokⅠgene polymorphism and type 2 diabetes incidence in the postmenopausal women in south Sichuan area.

OBJECTIVETo observe the effects of Shenqi Compound (SQC) on the mRNA expression of angiotensin II type 1 receptor (AT1R) in the aorta of Goto-Kakizaki (GK) rats.

METHODSTotally 67 GK rats were randomly divided into 5 groups, i.e., the GK group (n =18), the model group (n =16), the atorvastatin group (n =17), and the SQC group (n =16). Another a normal control group was set up (n =18). The diabetic macrovascular disease model was prepared by adding L-NAME (at the daily dose of 0.10 mg/mL) in drinking water for GK rats. GK rats, except those in the normal control group were fed with high fat diet. Atorvastatin (at the daily dose of 1.60 mg/kg) and SQC (at the daily dose of 1.44 g/kg) were respectively administered by gastrogavage, once daily for 35 successive days. The blood glucose was determined by glucose oxidase method once per week. After 5-week medication, the contents of triglyceride (TG) and total cholesterol (TC) were determined by ELISA. The serum concentrations of angiotensin I (Ang II) were determined by RIA. The mRNA expression of AT1R in the aorta was determined by real-time quantitative reverse transcriptase PCR (RT-PCR).

RESULTSThe blood glucose level was obviously lower in both the atorvastatin group and the SQC group after 4 weeks of medication (P <0.05). Besides, it was significantly lower in the SQC group than in the model group by the end of the 4th week (P <0.05). The concentrations of TG, TC and serum Ang II , and the mRNA expression of AT1R in the aorta were significantly higher in the model group than in the normal control group (P <0.01). After 5-week medication, the concentrations of TG, TC and serum Ang I , and the mRNA expression of AT1 R in the aorta were significantly lower in the atorvastatin group and the SQC group than in the model group (P <0.01, P <0.05). The mRNA expression of AT1R was significantly higher in the SQC group than in the atorvastatin group (P <0.05).

CONCLUSIONSSQC could significantly reduce the levels of blood glucose, TG, TC, down-regulate the mRNA expression of AT1R in the aorta, and decrease the expressions of serum Ang II of GK rats with diabetic macrovascular disease. AT1 R might be one of effective targets of SQC in treating diabetic macrovascular diseases.

Angiotensin II ; blood ; Animals ; Aorta ; drug effects ; metabolism ; Blood Glucose ; analysis ; Cholesterol ; blood ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; RNA, Messenger ; genetics ; Rats ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Triglycerides ; blood

Objective To observe the effects of culture medium of a mn iotic cells on NO and NOS in retinal tissues of rabbits in vitro in order to pro vide a protective method for antioxidation in retina transplantation. M ethods Thirty adult healthy rabbits (30 right eyes) were divided into 3 groups. Group Ⅰ: fresh retinal tissue; group Ⅱ: routine culture medium; group Ⅲ: culture medium of amniotic cells. The retinal tissues in group Ⅱ and Ⅲ we re cultured in the corresponding culture medium for 1 week. The content of NO an d NOS in retinal tissues in the 3 groups were determined. Results Compared with group I, the content of NO and NOS of group Ⅱ increased obvio usly (t=3.821, 3.854; P0.05) . Conclusion Culture medium of amniotic cells may remove free r adicals and enhance the ability of antioxidation.

BACKGROUNDHuman embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.

METHODSThree oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.

RESULTSFour ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.

CONCLUSIONSHES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.

Blastocyst ; cytology ; Cell Differentiation ; Cell Line ; DNA-Binding Proteins ; analysis ; Female ; Fertilization in Vitro ; Humans ; Karyotyping ; Male ; Octamer Transcription Factor-3 ; Stem Cells ; cytology ; Tissue Donors ; Transcription Factors ; analysis

OBJECTIVETo investigate the mechanism and factors affecting mosaicism in human preimplantation embryos by using 2 sequential rounds of fluorescence in situ hybridization(FISH).

METHODSTotally 51 normal fertilized embryos, which were not suitable for embryo transfer and cryopreservation, were analyzed on day 3 after fertilization by using two sequential rounds of FISH. Chromosomes 13, 16, 18, 21, 22, X and Y were analyzed.

RESULTSAmong 51 embryos, 16 (31.4%) were mosaic, 12 (23.5%) were chaotic, and the remaining were either normal (27.5%) or non-mosaic abnormal (17.6%). The incidence of mosaic embryos was related to embryo developmental stage, for the incidence of mosaicism increased from 12.5% in embryos

CONCLUSIONMosaicism is common in human preimplantaion embryos, which may be one of the important factors affecting the success rates in IVF-ET. Most of the chromosomal abnormalities can be identified by two sequential rounds of FISH.

Aneuploidy ; Blastocyst ; Chromosomes, Human ; Embryo Transfer ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Mosaicism ; chemically induced ; embryology ; Preimplantation Diagnosis

OBJECTIVEIn order to explore the roles of Huoxueruanjian compound on liver fibrogenesis and its molecular mechanism, this paper has investigated the Influence of blood serum with such traditional Chinese medicine compound on the expression of Smad3, Smad7 and procollagen alpha2(I) gene in hepatic stellate cell (HSC).

METHODSHSC-T6 was deal with different Concentration of blood serum medicine with Heluoshugan which was made by routine way. Then expression change of Smad3, Smad7 and procollagen alpha2(I) mRNA among each groups were observed by RT-PCR. Furthermore, the expression change of Smad3 protein were examined by Western blot.

RESULTSExpression of Smad3 and procollagen alpha2(I) mRNA as well as Smad3 protein had been downregulated after treating with blood serum medicine of Heluoshugan (P<0.01, P<0.05, respectively). The expression of procollagen alpha2(I) mRNA changed at the same tendency as those of Smad3. The role of blood serum medicine was significant difference between different concentration, P<0.05. And the expression of procollagen alpha2(I) mRNA changed in concentration-dependent manner. Blood serum medicine has no effects on the Smad7 mRNA.

CONCLUSIONThe anti-fibrosis roles of HuoXueruanjian Compound maybe influence the function of TGF-beta and Smad by nonspecific action, thereby inhibit the transcription of procollegan alpha2(I) mRNA and decrease the production of ECM. As regards Smad3, it may be facilitating the development of liver fibrosis when its expression increases. Otherwise, it manifest with anti-fibrosis role.

Animals ; Collagen Type I ; genetics ; DNA-Binding Proteins ; genetics ; Liver Cirrhosis ; drug therapy ; etiology ; pathology ; Medicine, Chinese Traditional ; RNA, Messenger ; analysis ; Rats ; Smad3 Protein ; Smad7 Protein ; Trans-Activators ; genetics

BACKGROUNDThe efficiency of traditional cryopreservation of human embryonic stem (ES) cells is low, and there have been few attempts to prove new cryopreservation methods effective. This study was designed to evaluate the efficiency of cryopreservation of human ES cells using vitrification method.

METHODSHuman ES cells clumped from an identical cell line were randomly allocated to be cryopreserved by vitrification or by slow freezing. The recovery rates, the growth and differentiation potential of thawed human ES cells were compared between these two groups. The pluripotency of human ES cells after thawing was identified.

RESULTSEighty-one point nine percent (59/72) of human ES cell clumps were recovered after vitrification, while only 22.8% (16/70) were recovered after slow freezing (P < 0.01). The colonies after vitrification manifested have not only faster growth but also a lower level of differentiation when compared to colonies subjected to the slow freezing protocol. However, the rates of growth and differentiation in undifferentiated colonies from both groups were identical to the rates in those of non-cryopreserved stem cells after a prolonged culture period. Passage 6 of vitrified human ES cells retained the properties of pluripotent cells, a normal karyotype and expressed the transcription factor OCT-4, stage specific expressed antigen-4 (SSEA-4) and SSEA-3. Teratoma growth of these cells demonstrated the ability to develop into all three germ layers.

CONCLUSIONSVitrification is effective in cryopreserving human ES cells. During a prolonged culture, human ES cells retain their pluripotency after cryopreservation.

Cell Differentiation ; Cell Survival ; Cryopreservation ; methods ; Embryo, Mammalian ; cytology ; Humans ; Osmotic Pressure ; Stem Cells ; cytology

OBJECTIVETo explore the relation between oocyte maturation and growth differentiation factor-9 (GDF-9) gene expression.

METHODSOvariectomy was performed in 50 Kunming female mice of 10 days old, and the preantral follicles were isolated from the ovaries and cultured in medium drops for 12 days. Oocytes and somatic cells were mechanically isolated. The oocytes cultured in vitro for 2, 4, 6, 8, 10, and 12 days constituted the in vitro cultured group and the oocytes obtained from female mice of 12, 14, 16, 18, 20, and 22 days old served as the in vivo group. Semi-quantitative RT-PCR and agar gel electrophoresis were performed to quantify GDF-9 gene expression in each oocyte.

RESULTSFollicle survival, antrum formation and maturation rate was 89.5%, 51.8% and 56.6% in the in vitro cultured follicles, respectively. GDF-9 gene expression on days 2, 4, 6, 8, 10, and 12 in in vitro cultured oocytes was 0.83-/+0.08, 0.52-/+0.09, 0.45-/+0.13, 0.49-/+0.09, 0.49-/+0.09, and 0.68-/+0.08, respectively; GDF-9 gene expression in in vivo grown oocytes of 12, 14, 16, 18, 20, and 22 days were 0.64-/+0.35, 0.48-/+0.10, 0.52-/+0.10, 0.66-/+0.08, 0.72-/+0.09, and 0.91-/+0.11, respectively. Between days 8 and 12, GDF-9 gene expression in in vitro cultured oocyte was significantly lower than that in in vivo grown oocytes (P<0.05).

CONCLUSIONMII oocytes can be obtained from in vitro culture of the preantral follicles. GDF-9 gene expression in the oocytes varies with their growth stages. Between days 8 and 12 of in vitro culture, GDF-9 gene expression in the cultured oocytes is different from that in in vivo grown oocytes.

Animals ; Animals, Newborn ; Bone Morphogenetic Protein 15 ; Cell Survival ; Cells, Cultured ; Electrophoresis, Agar Gel ; Female ; Gene Expression ; Growth Differentiation Factor 9 ; Intercellular Signaling Peptides and Proteins ; genetics ; Mice ; Oocytes ; cytology ; growth & development ; metabolism ; Ovarian Follicle ; cytology ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors