Objective To test whether feeder cells derived from mouse embryonic stem cells(mESCs) could support the growth of mESCs themselves. Methods mESCs were induced to form mouse embryoid bodies(EB),and then fibroblast-like cells were derived from further differentiated mEB(mEB-dF),which served as feeder cells.The undifferentiation of mESCs grown on mEB-dF was confirmed by morphological analysis,colony efficiency and cell differentiation rate of mESCs,immunocytochemistry,alkaline phosphatase staining and RT-PCR.The pluripotency of mESCs grown on mEB-dF was examined by RT-PCR,inducing their differentiation in vivo and in vitro. Results(Forty-eight) fibroblast-like cells lines were derived from the same EB at three periods(d 10,d 15 and d 20),and five of them,mostly derived from d15 EB,were able to maintain mESCs in undifferentiated status and pluripotential ability over 10 passages.mESCs cultured on these feeder cell lines expressed alkaline phosphatase and specific mESCs markers,including SSEA-1,OCT-4,NANOG,and formed EB in vitro and teratomas in vivo.However,the majority of mEB-dF lines(43/48) has no such ability. Conclusion This study not only provides a novel feeder system for mESCs culture,avoiding lot of disadvantages of mouse embryo fibroblasts used as the feeder,but also indicates that fibroblast-like cells derived from mESCs take on different functions.The molecular mechanism of different function of these fibroblast cells is worthy of further investigations.

Objective To observe the clinical effects of hyaluronidase iontophoresis in the treatment of in- fants with congenital muscular torticollis(CMT).Methods Fifty infants with congenital muscular torticollis were divided randomly into a treatment group and a control group.Manual stretching was performed with both groups,while hyaluronidase iontophoresis was also administered to those in the treatment group.The range of side-flexion and rota- tion of the neck was examined at the beginning of and after 1 and 2 months of treatment.At the end of the treatment, the overall outcome was assessed according to a scoring system.Results There was no significant difference be- tween the two groups at admission.Compared with the control group,the range of side-flexion and rotation of the neck,and the overall treatment outcome were all significantly better in the treatment group after treatment.Conclu- sion Hyaluronidase iontophoresis can effectively improve function in infants with CMT and alleviate their symptoms.

Objective:To investigate the relationship between clinical effects of acupuncture for elderly patients with sensorineural deafness and ear distending sensation.Methods:A total of 120 elderly patients with sensorineural deafness were randomly divided into a comprehensive treatment group,an acupuncture group and a Western medicine group,with 40 cases in each group.The acupuncture group received acupuncture treatment,the Western medicine group received oral mecobalamin tablets and the comprehensive treatment group received acupuncture plus acupoint injection and auricular acupoint sticking.The values of pure tone hearing threshold test of the three groups were observed before and after treatment,and the relationship between clinical effects and ear distending sensation was compared.Results:The total effective rate of the comprehensive treatment group was 82.5％ versus 67.0％ in the acupuncture group and 62.5％ in the Western medicine group.The inter-group comparisons showed statistically significant differences in the comprehensive treatment group versus the Western medicine group (P＜0.01) and the acupuncture group (P＜0.05).In the comprehensive treatment group,there were 23 cases (57.5％) with ear distending sensation,the clinical total effective rate was 86.9％;there were 17 cases (42.5％) without the sensation,the clinical total effective rate was 76.5％.In the acupuncture group,there were 24 cases (60.0％) with ear distending sensation,the clinical total effective rate was 71.0％;there were 16 cases (40.0％) without the sensation,the clinical total effective rate was 63.0％.In the Western medicine group,there were 21 cases (52.5％) with ear distending sensation,the clinical total effective rate was 66.7％;there were 19 cases (47.5％) without the sensation,the clinical total effective rate was 57.9％.The total effective rate of patients with ear distending sensation were higher than the rates of those without ear distending sensation in the three groups,but the differences were insignificant (all P＞0.05).Conclusion:The comprehensive therapy is one of the effective methods to treat elderly patients with sensorineural deafness.In the three groups of elderly patients with sensorineural deafness,the relief of ear distending sensation and the hearing loss were basically simultaneous,and the hearing recovery in the patients with ear distending sensation may be slightly better than that in those without ear distending sensation.Nevertheless,further research is needed.

Background:Gastric cancer (GC) is one of the most common malignancies,and intestinal-type GC is the main histopathologic type of GC in China.We previously reported that casein kinase 2 interacting protein 1 (CKIP-1) acts as a candidate tumor suppressor in intestinal-type GC.CKIP-1 participates in the regulation of multiple signaling pathways,including the Wnt/β-catenin pathway,of which caudal-related homeobox 1 (CDX1) may be a downstream target gene.The purpose of this study was to investigate the relationship between CKIP-1 and CDX1 in intestinal-type GC.Methods:Sixty-seven gastroscopy biopsy specimens and surgically resected gastric specimens were divided into four groups:gastric mucosa group,intestinal metaplasia (IM) group,dysplasia group,and intestinal-type GC group.The expression levels of CKIP-1 and CDX1 were detected in these groups and GC cell lines,and the correlations between these expression levels were analyzed.SGC7901 and BGC823 cells were divided into CKIP-1 shRNA groups and CKIP-1 over-expression groups,and CDX1 expression was detected.β-Catenin expression was detected in intestinal-type GC tissue samples and CKIP-1 shRNA and CKIP-1 over-expression SGC7901 cells,and its correlation with CKIP-1 expression in intestinal-type GC tissue was analyzed.The Wnt/β-catenin pathway inhibitor DKK-1 and activator LiCl were incubated with SGC7901 cells,BGC823 cells,and CKIP-1 shRNA and CKIP-1 over-expression SGC7901 and BGC823 cells,following which CDX1 and Ki-67 expression were detected.Results:The expression levels of CKIP-1 and CDX1 were lower in patients with intestinal-type GC than in patients with IM and dysplasia (both P ＜ 0.05).CKIP-1 and CDX1 expression levels were positively correlated in IM,dysplasia,and intestinal-type GC tissue and cell lines (r =0.771,P ＜ 0.01;r =0.597,P ＜ 0.01;r =0.654,P ＜ 0.01;r =0.811,P ＜ 0.01,respectively).CDX1 expression was decreased in the CKIP-1 shRNA groups and increased in the CKIP-1 over-expression groups of SGC7901 and BGC823 cells compared to that in the corresponding control groups (both P ＜ 0.05).CKIP-1 expression was negatively correlated with β-catenin expression in intestinal-type GC patients (r =-0.458,P ＜ 0.01).Compared to the control group,β-catenin expression was increased in the CKIP-1 shRNA SGC7901 cell group and decreased in the CKIP-1 over-expression SGC7901 cell group (P ＜ 0.05).CDX1 expression was increased in SGC7901 and BGC823 cells treated with DKK-1,DKK-1 increased CDX1 expression and decreased Ki-67 expression in the CKIP-1 shRNA group;the opposite result was observed in SGC7901 and BGC823 cells treated with LiCl,and LiCl decreased CDX1 expression and increased Ki-67 expression in the CKIP-1 over-expression group (both P ＜ 0.05).Conclusions:Through the Wnt/p-catenin signaling pathway,CKIP-1 may positively regulate CDX1 in intestinal-type GC.

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

OBJECTIVETo investigate the effects and mechanisms of high glucose on the phenotype transformation of rat vascular smooth muscle cells (VSMCs).

METHODSVSMCs ere isolated from rat thoracic aorta and the 3rd-5th VSMCs were incubated with normal glucose (5.5 mmol/L), high glucose (25 mmol/L), or high glucose (25 mmol/L) + P38 inhibitor (25 mmol/L +SB203580) for another 24 hours. Then the gene expression of osteopontin (OPN), alpha smooth-actin (alpha-SMA), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9(MMP-9) were assayed by real time RT-PCR, the protein expression of P38 were assayed by Western blot.

RESULTS(1) High glucose promoted the phenotype transformation of VSMCs and up-regulated the expression of MMP-2 and MMP-9. (2) High glucose promoted the phosphorylation of P38. (3) SB203580, the inhibitor of P38/MAPK signal pathway, inhibited the effects of high glucose on phenotype transformation and expression of MMP-2 and MMP-9.

CONCLUSIONHigh glucose may promote phenotype transformation of VSMCs via the signal pathway of P38/MAPK.

Actins ; metabolism ; Animals ; Aorta, Thoracic ; cytology ; Blotting, Western ; Cells, Cultured ; Glucose ; pharmacology ; Imidazoles ; pharmacology ; MAP Kinase Signaling System ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Osteopontin ; metabolism ; Phenotype ; Pyridines ; pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases ; metabolism

Adolescent ; Adolescent Development ; Child ; Child Development ; China ; epidemiology ; Female ; Humans ; Male ; Obesity ; epidemiology ; Prevalence

OBJECTIVETo investigate whether atrial expression of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) of right atrial appendages are altered in patients with rheumatic valvular disease during chronic atrial fibrillation.

METHODSA total of 48 patients with rheumatic heart disease were included. 27 patients had no history of atrial fibrillation, 21 patients had atrial fibrillation. Atrial tissue was obtained from the right atrial appendage during open heart surgery. The protein expression of IL-1beta and TNF-alpha was detected by immunohistochemistry method. The fibrosis of right atrial appendage was detected by Masson staining.

RESULTSThe fibrosis of right atrial appendage was significantly increased in patients with chronic atrial fibrillation. The protein expression of IL-1beta and TNF-alpha were significantly increased in patients with chronic atrial fibrillation.

CONCLUSIONSThe protein expression of IL-1beta and TNF-alpha were significantly increased in patients with rheumatic valvular disease during chronic atrial fibrillation. Inflammation may be one of the mechanisms for the development and persistence of atrial fibrillation.

Adult ; Aged ; Atrial Fibrillation ; complications ; metabolism ; Female ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Rheumatic Heart Disease ; complications ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.
Biomarkers, Tumor ; Breast Neoplasms ; genetics ; metabolism ; Cell Movement ; genetics ; Epithelial-Mesenchymal Transition ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; HMGB1 Protein ; genetics ; Homeodomain Proteins ; biosynthesis ; Humans ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; genetics ; Neoplasm Metastasis ; genetics ; pathology ; Repressor Proteins ; biosynthesis ; Transcription Factors ; biosynthesis ; Zinc Finger E-box Binding Homeobox 2 ; Zinc Finger E-box-Binding Homeobox 1

Objective To study the prevalence and characteristics of blood pressure (BP) among children and adolescents at normal weight but with abdominal obesity.Methods Using data from the ‘Student physical fitness and health surveillance 2010 project' in Shandong province,a total of 38 816 students aged 7-17 years were selected to participate in this study.Stature,body weight,waist circumference (WC),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of these subjects were measured.Body weight status and abdominal obesity were defined by body mass index (BMI) and WC,respectively.Results In total,the proportions of thinness,normal weight,overweight and obesity defined by BMI were 5.37％,72.47％,12.92％ and 9.24％ respectively.5.86％ of the children and adolescents with normal weight had abdominal obesity,with normal weighted girls (7.19％) having higher prevalence of abdominal obesity than boys (4.33％) (P＜0.01).The Z-scores of SBP and DBP for both boys and girls were all significantly higher in the normal weight but with abdominal obesity groups than in both normal weight and WC groups (P＜0.01).Conclusion Children and adolescents under normal weight but with abdominal obesity had higher BP level need to be identified and considered as high-risk individuals.Related intervention programs should also be targeted to this population.