Objective To explore the correlation between the amount of residual undifferentiated embryonic stem cells (ESCs) in embryoid bodies and its tumorigenicity.Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies (EBs).Ten days later,EBs were digested into single cells and then re-plated in standard ESCs culture condition.The residual undifferentiated embryonic stem cells surface maker SSEA-1 was examined by flow cytometry in EBs.The morphology of residual undifferentiated cells in EBs were observed,meanwhile the surface marker SSEA-1 was examined by immunofluorescent staining.EBs were digested into single cells and grouped into 104,105,106,2×106,and then injected into limb muscle of nude mice.The correlation of the amount of cells and its tumorigenicity was observed.Results Residual undifferentiated ESCs were observed after EBs differentiated for 10 days,which displayed clonal morphology and expressed undifferentiated ceil markers of ESCs,such as SSEA-1.The expression rate of undifferentiated cells surface marker SSEA-1 was (13.5±0.75)％ in EBs differentiated for 10 days.Only two millions single cells harvested from EBs were able to form teratoma after being injected into muscle of nude mice for 6 weeks.Mature endoderm,mesoderm and ectoderm tissues could be found in teratoma.No teratoma formed in other groups.Conclusion A certain amount residual undifferentiated ESCs still exist after differentiation of ESCs into EBs.About 2.7× 105 undifferentiated cells are able to form teratoma by iniecting into muscle of nude mice.

Objective:To observe the theraputic effect and changes of glomerular extracellular matrix components(ECM) and PAI-1 and their relationships after cure with methyl-prednisone on immune complexes nephritis rats.Methods:Immune complexes nephritis rats model were induced with C-BSA.Levels of FN,LN in different treatment groups were analyzed by ELISA, level of PAI-1 in rats renal tissue was determined by color developing substrate,was theraputic effect of methyl-prednisone with 24 hours volumes of urine protein.Results:The levels of PAI-1,FN and LN of model groups were significantly higher than those of normal and control groups,and PAI-1 was significantly correlated with LN and FN,Glomerular mesangial matrix proliferated slightly and moderately;The levels of FN,LN and PAI-1 decreased signifcantly and glomerular mesangial matrix proliferation lessen differently after cure of methyl-prednisone for 1 and 2 weeks,but not reaching the level of normal groups;24 hours volumes of urine protein decrease significantly in treatment groups.Conclusion:The ECM accumulation correlate with PAI-1 increase in immune complexes rats,methyl-prednisone may affect ECM accumulation by interfering with PA/PAI-1 system to reach a treatment purpose.

Objective: To prepare tissue engeneered bone in the shape of human TMJ condyle. Methods: Rabbit marrow stem cells (MSCs) were in vitro cultured and induced by rhBMP2.107 Cells were seeded into each piece of natural porous coral (NC) in the shape and in the size of 4 -year-old-child mandibular condyle. After two days in vitro incubation, six cell-coral complexes were implanted subcautanrously into the back of nude mice. Two months after operation, bone formation was observed by gross inspection,X-ray examination,scanning electronic microscope observation and histological observation. Results: New bone grafts in the shape of human mandibular condyle were successfully restored two months after implantation in all the samples. X-ray examination showed large amount of X-ray blocking shadow. NC was partially absorbed. New bone formation could be observed by electronic microscope observation and hostological observation on the surface and in the pores of NC. Conclusion: It is an effective method to fabricate bone graft in specific shape by seeding osteogenesis cells into natural coral in the wanted shape.

Objective: To prepare tissue engieered bone graft loading titanium dental implant. Methods: Titanium dental implant (3 mm in diameter) was inserted into porous natural coral column((5 mm in diameter). Bone marrow derived osteoblasts were cultured and expanded in vitro. Cells were induced by recombinant human bone morphogenetic protein-2 for three days and then harvested and seeded into porous coral and onto dental implant at the density of 2 ?108/ml. Four cell-coral-implant complexs were incubated in vitro for 2 days and then implanted subcutaniously into nude mice. New bone formationre and new bone integration with dental implant were evaluated by gross inspection, X-ray examination and hitologic observation 1 and 2 months after implantation. Results: By gross observation, specimen of 1 month was red and white. X-ray examination showed that there was little radiodense shadow around the dental implant. Specimen of 2 months was red and had the gross appearance of bone. Dental implant could be observed situating in the newly formed bone graft. X-ray examination showed that coral scaffold was absorbed completely. Large amount of X -ray blocking shadow could be observed around the dental implant. Histologic examination showed that bone-like tissue formed in the pores and on the surface of natural coral and in some area new bone could be observed integrating with implant in 1 month specimen. In 2 months specimen, large amount of new bone formed around the implant and integrated well with the implant. Conclusions: Tissue engineered bone graft may integrate well with titatium dental implant.

OBJECTIVETo investigate the feasibility of using marrow stromal osteoblast (MSO) as bone derived cell and using cancellous bone matrix (CBM) as scaffold for bone tissue engineering, the subcutaneous osteogenesis of MSO-CBM compound artificial bone (MCCAB) was observed in the experiment.

METHODSThe marrow stromal cells of adult New Zealand rabbits cultivated and induced in vitro were used to form MCCAB by mixing, seeding and solidifying methods assisted by alginate. The MCCABs were auto-transplanted subcutaneously into the rabbits for 4 to 8 weeks. The alginate-cancellous bone matrix composites or the cancellous bone matrix alone were implanted as control. The effectiveness of bone formation was assessed by means of roentgenography, histology and computerized histomorphometry.

RESULTSThe osteogenesis of MCCABs was better than that of the alginate-cancellous bone matrix composites and of the cancellous bone matrixes. In the MCCABs, both intramembranous and cartilaginous osteogeneses were seen but the former was obvious. In the control, only slight cartilaginous osteogeneses were seen.

CONCLUSIONSThe osteogeneses of the MCCABs constructed by using tissue engineering method were obvious when transplanted subcutaneously. The MSO and CBM can be used as good bone-derived cell and scaffold material respectively for tissue-engineered bone construction.

Animals ; Bone Marrow Transplantation ; Bone Matrix ; transplantation ; Bone Transplantation ; methods ; Male ; Osteoblasts ; transplantation ; Osteogenesis ; Rabbits ; Tissue Engineering

OBJECTIVETo investigate the feasibility of using marrow stromal osteoblast-cancellous bone matrix compound artificial bone (MCCAB) as tissue-engineered bone, the osteogenesis of MCCAB in the cranial defect was observed in the experiment.

METHODSThe in vitro cultivated and induced marrow stromal cells of adult New Zealand rabbits were seeded into the alginate-cancellous bone matrix to form MCCAB. The MCCAB was then implanted into the cranial defect for 4 to 8 weeks. The cancellous bone matrix (CBM) alone or the marrow stromal osteoblasts (MSOs) alone was implanted as the control. The effectiveness of bone formation was assessed by histological and roentgenographic analysis.

RESULTSThe osteogenesis of MCCAB was better than CBM or MSOs and superior to the blank group.

CONCLUSIONMCCAB can effectively repair cranial defect. It could be used clinically to restore large bone defects.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Matrix ; cytology ; Cells, Cultured ; Feasibility Studies ; Male ; Osteoblasts ; cytology ; physiology ; Rabbits ; Skull ; abnormalities ; Stromal Cells ; cytology ; physiology

OBJECTIVETo investigate the therapeutical effects of visfatin and metformin on insulin resistance and reproductive endocrine disorder in rats with polycystic ovary syndrome (PCOS).

METHODSForty female Wistar rats were divided into 4 equal groups, and in groups A, B and C, the rats were injected subcutaneously with dehydroepiandrosterone (DHEA) for PCOS modeling, with group D as the blank control injected with soybean oil. Vaginal smears and serological testing were taken to assess the modeling. After the modeling, the rats in group A received 10 µg reorganized visfatin injection and those in group B were treated with metformin (14 mg/100 g) on a daily basis for 15 days. Serum levels of T, LH, FSH, FINS and blood glucose levels during OGTT were measured before and after the treatments, and HOMA-IR and LH to FSH ratio were calculated. The ovaries were then dissected for pathological examination.

RESULTSIn groups A and B, FINS, FPG, T, HOMA-IR and blood glucose levels during OGTT were significantly decreased after the treatments (P<0.05), which resulted in recovery of regular menses in 8 (80%) rats in group A and 7 (77.8%) rats in group B with the development of normal follicles. Visfatin and metformin produced equivalent therapeutic effects in improving the insulin resistance and hyperandrogenism in PCOS rats.

CONCLUSIONVisfatin and metformin have equivalent therapeutic effects in improving insulin resistance and hyperandrogenism and in promoting the recovery of regular menses and development of normal follicles in PCOS rats.

Animals ; Female ; Humans ; Insulin Resistance ; Metformin ; pharmacology ; Nicotinamide Phosphoribosyltransferase ; pharmacology ; Polycystic Ovary Syndrome ; complications ; drug therapy ; Rats ; Rats, Wistar

BACKGROUND AND OBJECTIVEThe relationship between myeloperoxidase G-463A genetic polymorphisms and lung cancer susceptibility has been studied extensively. However, the outcomes are not consistent. The aim of this study is to evaluate the relationship between myeloperoxidase genetic polymorphisms and lung cancer susceptibility by meta analysis.

METHODSDocuments published were retrieved through databases associated with the study. Taking into account the possibilities of heterogeneity of the studies, a statistical test for heterngeneity was performed. The odds ratio and 95% CI were used to evaluate the risks. The meta analysis was applied with RevMan software 4.2, and the forest plot and funnel plot of meta analysis were worked out.

RESULTSA total of 5 381 cases and 5 827 controls from studies for Caucasian and a total of 1 558 cases and 1 755 controls from studies for East Asians were included. For Caucasian the pooled OR was 0.91 (95% CI: 0.81-1.02); For East Asians, the pooled OR is 0.83 (95% CI: 0.63-1.09). Publication bias exits in the study for Caucasian, but not for East Asians.

CONCLUSIONThe results of this study indicated that the polymorphism of myeloperoxidase G-463A was not significantly associated with the lung cancer risk for Caucasian or East Asians. However, further studies for the East Asians is needed for the few subjects.

Genetic Predisposition to Disease ; genetics ; Humans ; Lung Neoplasms ; epidemiology ; genetics ; Peroxidase ; genetics ; Polymorphism, Genetic ; genetics

Objective To investigate the association between ten single nucleotide polymorphisms (SNPs) in the peroxisome proliferator-activated receptors and pulse pressure (PP) as well as the relationships between gene-gene interaction between PPARα/δ/γ genes and PP.Methods A total of 820 subjects,with 550 females and 270 males,were recruited from a cohort study of “Prevention of Metabolic Syndrome and Multi-metabolic Disorders in Jiangsu Province of China Study (PMMJS)”.Ten SNPs of PPARα/δ/γ genes were selected.GMDR software (version 1.0.1) was used to evaluate the gene-gene interactions among PPARs SNPs associated with PP.Results The mean levels of PP in people with mutant genotype of rs1805192 in PPARγ genes (PA+AA) showed a significant increase by 1.341 mmHg (95％CI:0.431-2.252 mmHg) when compared to the persons with wild genotype (PP).In the subgroup of subjects with more than 30 mmHg levels of PP,a six-locus model comprised rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγshowed a highest level of prediction accuracy (0.577) and displayed a better cross-validation consistency (10/10).In the subgroup of subjects with less than 40 mmHg levels of PP,a two-locus model was statistically associated with PP with 0.628 of prediction accuracy and 10/10 of cross-validation consistency.Conclusion PPARγrs1805192 was associated with the occurrence of PP.Gene-gene interactions among rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγ were all significantly related to PP.

Objective:To investigate the association between plasma anti-Müllerian hormone(AMH) levels and ischemic stroke.Methods:In this case-control study, 93 ischemic stroke patients were randomly selected as the case group from a study on the prevention and treatment of metabolic syndrome, which was conducted in 2018-2019 in Changshu, Jiangsu Province, while 372 nonischemic stroke patients were selected as the control group according to the principle of 1∶4 matching.An enzyme-linked immunosorbent assay was used to measure plasma AMH levels.The conditional logistic regression model and restricted cubic spline were used to analyze the relationship between AMH levels and ischemic stroke.Results:A total of 465 subjects with an average age of (68.7±7.4)years were included in this study, of whom 215(46.2%)were men and 250(53.8%)were women.According to our conditional Logistic regression analysis, the risk of ischemic stroke was reduced by 44% for every unit increase in the log-AMH level( OR=0.56, 95% CI: 0.37-0.85)in the overall population after multivariate adjustment.Compared with the tertile with the lowest AMH level, the risk of ischemic stroke in the tertile with the highest AMH level decreased significantly( OR=0.37, 95% CI: 0.19-0.69). When subgrouped by sex, the tertiles with the highest AMH levels were associated with a 66% lower risk of ischemic stroke in men( OR=0.34, 95% CI: 0.13-0.88)and a 64% lower risk of ischemic stroke in women( OR=0.36, 95% CI: 0.15-0.87), compared with the tertiles with the lowest AMH levels.The results of restricted cubic spline analysis showed that there was a linear dose-response relationship between plasma AMH levels and ischemic stroke both in the general population and in male or female population( Pvalues for linear trends were 0.0002, 0.008 and 0.007, respectively). Conclusions:Higher plasma AMH levels decrease the risk of ischemic stroke with a dose-response pattern.