Bone marrow stem cells(BMSCs) can differentiate into cardiomyocytes when induced with proper reagent. The cardiomyocytes from BMSCs have cardiomyocyte phenotypes with the expression of cardiac-specific antigen and receptor. BMSCs differentiate into cardiomyocytes in host myocardial microenvironments. Mechanism of the differentiation is unclear and need to be studied further.

Objective To investigate the role of bone marrow mesenchymal stem cells (MSCs) transplantation in repairing injured intestinal mucosa of acute pancreatitis.Methods MSCs were harvested and cultured from femurs of male SD rats.Twenty female SD rats were divided into three groups,and serve acute pancratitis (SAP) model was induced by intraperitoneal injection of L-arginine (2 g/kg) twice.Twelve hours after SAP model established,MSC transplantation group (n=8) were injected MSCs (5 × 106 cell/rat) through tail vein for three days,and SAP group (n=6) were injected the same volume of saline through tail vein as control.Control group (n=6) were only injected the same volume of saline without any treatment.All the rats were sacrificed at 72 hours after model established.The small intestinal tissues were taken for HE staining and pathological score,the TNF-α mRNA and IL-1β mRNA expression level in small intestine and pancreas were tested by RT-PCR.Y chromo-some (Sry) gene in pancreatic and intestinal tissue was examined by polymerase chain reaction (PCR).Results The relative expression quantity of TNF-a mRNA and IL-1β mRNA in pancreas was significant higher in SAP group and MSC transplantation group than in control group (7.22 ± 1.99,3.46± 1.75 vs 1.32 ± 1.04 ; 2.71 ± 0.56,1.92 ± 0.28 vs 0.61 ± 0.45 ),the difference was statistically significant (F=18.375,F=22.701; P＜0.05).Compared with SAP group,the expression quantity of TNF-α mRNA and IL-1β mRNA in pancreas was significantly decreased in MSC transplantation group,the difference was statistically significant (P＜0.05).The relative expression quantity of TNF-α mRNA and IL-1β mRNA in small intestine was significantly higher in SAP group and MSC transplantation group than in control group (3.93 ± 1.08,2.13 ± 0.53 vs 0.68 ± 0.42 ; 2.44 ± 1.54,1.02±0.44 vs 0.60±0.14),the difference was statistically significant (F=21.772,F=6.132; P＜0.05).The expression of TNF-αmRNA and IL-1β mRNA in MSC transplantation group was lower than that in SAP group,the difference was statistically significant (P＜0.05).Compared with SAP group,pathological score indicated that small intestine injure was slighter in MSC transplantation group (3.83±0.28 vs 2.83±0.56),the difference was statistically significant (F=12.013,P＜0.05).Sry gene could be detected in the pancreatic and intestinal tissue of MSC transplantation group.Conclusion Allogeneic MSC transplantation group can inhibit Pro-inflammatory cytokines expression in acute pancreatitis,relieve the intestinal mucosa injury and may involve in the intestinal tissue repair.

Purpose To prepare streptavidin-tagged mouse interleukin-4(mIL-4-SA)bifunctional fusion protein and to study on its bioactivity.Methods The mIL-4 gene was cloned by RT-PCR and cloned into pET21 vector to get mIL-4-SA-pET21 expression plasmid.The mIL-4-SA fusion protein was expressed in BL21 (DE3)host bacteria and purified through the Ni-NTA affinity chromatography and refolded by dilution and dialysis.The effect of mIL-4-SA fusion protein on mouse thymocytes proliferation was evaluated by MTY.Flow cytometric analysis was performed to detect the mIL-4-SA fusion protein on the biotinylated B16F10 tumor cells.Results The mIL-4-SA-pET21 vector was successful by constructed and the mIL-4-SA fusion protein was expressed in BL21(DE3)at about 35%of total bacterial proteins.The purity of mIL-4-SA Was about 95% through Ni-NTA.The mIL-4-SA fusion protein exhibited bifunctional activities,i.e.,stimulative effect for mouse thymocyte proliferation and SA-mediated high-affinity binding to biotinylated cell surfaces(anchoring modified rate Was about 96.69%).Conclusion The mIL-4-SA fusion protein was expected to be developed for the treatment of tumors.

Objective:To study the effects of combination of rhubarb and early enteral nutrition on severe acute pancreatitis(SAP).Methods:A total of 73 patients with SAP were randomly divided into one of the three groups: 21 patients treated with total parenteral nutrition(TPN),27 patients treated with early enteral nutrition(EEN),and 25 patients treated with combination of rhubarb and early enteral nutrition(REN).The hospitalization cost,length of stay,mortality,recovery time of intestinal function and score of APACHE Ⅱ were observed.The plasma procalcitonin(PCT),the levels of C-reactive protein(CRP)and prealbumin were detected.Results: In REN group,there was no death.There was 1 patient died in EEN group and 2 patients in TPN group.The recovery time of intestinal function in REN group was(2.49?0.59) days,the length of stay was(25.13?4.35) days,and the hospitalization cost was(56.9?7.2) thousand yuan.They were obviously shorter than those in the other groups(TPN group,P

Objective To investigate the distribution of HLA-B27 subtypes in ankylosing spondylitis(AS) patients of Chinese Han population by using the updated HLA-B27 typing data. Methods One hundred AS subjects were randomly selected from spondyloarthritis patients data bank of the third affiliated hospital of Sun Yat-sen university. All subjects were independent individuals, and the duplicated samples in the same family were excluded. Salt fraction method was used to prepare genome DNA. Luminex liquid array combining PCR-SSOP was used to perform the low resolution HLA-B genotyping. PCR-SSP was applied to perform the high resolution HLA-B27 typing for HLA-B27 positive subjects. Results Ninety-eight independent AS patients were recruited randomily, of which, 93 were HLA-B27 positive, with positive rate 94.9%, and covered 96% patients with family history of AS. Three subtypes were detected in this population including B * 2704 (n=76, 81.7%), B * 2705 (n=12, 12.9%) and B * 2715 (n=5, 5.4%). Compared with the two reports about HLA-B27 subtype distribution in healthy HLA-B27 positive Han population there was no significant difference between AS patients and healthy controls. But no B * 2715 case was found in those two reports of healthy population. Three reports (including 1 report in Chinese) could found about B * 2715 subtype, but all positive cases were oriental people. Furthermore, all B * 2715 positive patients were AS patients. Conclusion B * 2704 is the predominant subtype ,in AS patients of Chinese Han population, and followed by B * 2705. We found five cases with positive B * 2715, a considerable rare allele. This may suggest association between B * 2715 and AS.

Objective To explore the effects of exclusive enteral nutrition (EEN) on the mucosal healing under endoscopy in adults with active Crohn's disease (CD).Methods From August 2011 to August 2012,adults with active CD met the inclusion criteria were prospectively enrolled and treated with EEN for four weeks.CD activity index(CDAI) score,C-reactive protein (CRP) were evaluated once enrolled and by the end of the second and fourth week.Colonoscopy or small intestinal endoscopy examination was conducted once enrolled and by the end of fourth week,and CD simplified endoscopic score (SES-CD) was also caculated.Wilcoxon test was applied to compare the efficacy.Results Eight patients met inclusion criteria and finished the study.Seven cases achieved clinical remission.Compared with those before treatment,both CDAI score and CRP level decreased significantly after treatment (both Z=2.52,both P=0.012).After treatment,two patients achieved mucosal healing and the mucosa of six cases improved.The mucosa healing rate and improvement rate of terminal ileum was higher than that of colon (3/8 vs 1/6; 5/8 vs 3/6).Compared with those before treatment,SES-CD total score and SES-CD score of ileum decreased significantly after treatment (Z=2.23,2.07; P=0.026,0.038).There was no significant difference in SES-CD score of colon before and after treatment (P =0.102).Conclusions EEN could induce clinical remission in adults with active CD,and promote the healing of mucosal ulcer under endoscopy.The healing rate of colon was lower than that of small intestine.

Background and purpose: As one of the important epigenetic phenomena, DNA methylation plays an important regulatory function for the expression of genes. Study shows that abnormal changes of DNA methy-lation patterns of normal tumor cell genome leads to dysfunction of cancer related gene, and this may be associated with tumor occurrence and development. The study investigated the promoter methylation and expression of caveolin-1 (CAV-1) gene in esophageal squamous cell carcinoma (ESCC), and to elucidate its role in ESCC. Methods:We used MSP approach, RT-PCR, and immunohistochemistry method respectively to examine the methylation status of the 5’CpG island of CAV-1 gene and its expression at mRNA and protein levels in tumors and corresponding normal tissues. Results: CAV-1 mRNA expression in tumor tissues (0.86±0.56) was signiifcantly higher than that in corresponding normal tissues (0.40±0.36, P<0.05). The mRNA expression of CAV-1 was correlated with status of lymphatic metastasis and TNM stage of ESCC patients (P<0.05). The protein expression of CAV-1 in tumor specimens (66.7%, 34/51) was signiifcantly higher than that in corresponding normal tissues (15.7%, 8/51, P<0.01). The protein expression of CAV-1 was signiifcantly associated with lymphatic metastasis of ESCC (P<0.05), however, it was not associated with differen-tiation and TNM stage (P>0.05). The promoter methylation frequency of CAV-1 in tumor specimens was 2.0%(1/51), and the methylation phenomenon has not been found in corresponding normal tissues. The promoter methylation fre-quency of CAV-1 in tumor specimens showed no signiifcant difference compared with the corresponding normal tissues (P>0.05). Conclusion:The mRNA and protein expression of CAV-1 in tumor specimens was signiifcantly higher than that in corresponding normal tissues. Aberrant high expression of CAV-1 has played a certain role in promoting tumori-genesis and lymph node metastasis. The expression both in ESCC and corresponding normal tissues has no correlation with the promoter methylation status.

Purpose To investigate the expression of Cav-1 in gastric cancer ( GC) cell lines and GC samples, and to analyze the pos-sible effect of gene methylation in expression of Cav-1. Methods Methylation specific PCR ( MSP) method was applied to examine the CpG methylation of the Cav-1 promoter in GC cell lines (AGS, MKN45, BGC-823) and 104 samples of GC and corresponding ad-jacent tissues. RT-PCR method was applied to examine the mRNA expression in GC cell lines. IHC were applied to examine the pro-tein expression of Cav-1 in the GC tissues. Results The expression level of Cav-1 mRNA was obviously increased after treated with 5-aza-2’-deoxycytidine (5-Aza-Dc, a demethylation agent) in AGS cell line. We detected the positive expression of Cav-1 gene in MKN45 and BGC-823 cell lines before and after treated with 5-Aza-Dc. The level of Cav-1 mRNA expression was no any change in AGS, MKN45 and BGC-823 cell lines treated with trichostatin A (TSA). MSP results showed that it can be amplified methylated bands in AGS cell line, and the methylated bands disappeared after treated with 5-Aza-Dc. MKN45 and BGC-823 cell lines were no any methylated bands amplified before and after treatment. The methylation frequency of Cav-1 gene was 29. 8% (31/104), which was significantly higher than that in adjacent tissues (P=0. 000). Furthermore, Cav-1 gene hypermethylation status was correlated with lymph node metastasis and family history of upper gastrointestinal cancers ( UGIC) , but not with pathological grade and clinical stage (P>0. 05). The positive frequency of Cav-1 expression was 51. 9%(54/104) in GC, which was significantly lower than that in adja-cent tissues (P=0. 000). The expression of Cav-1 was correlated with the frequency of gene methylation in GC tissues (P=0. 000). Conclusion The expression of Cav-1 was reduced in GC tissues and the gene hyermethylation may be one of the mechanisms causing Cav-1 gene silencing.

AIM:To explore the changes of plasma levels of soluble vascular endothelial growth factor receptor 2 ( sVEGFR2) and superoxide dismutase ( SOD) in hypertensive patients and hypertensive diabetic patients.METHODS:In this cross-sectional study, 88 cases were enrolled, which were divided into hypertensive group (n=31), hypertensive diabetic group ( n=31 ) and control group ( n=26 ) .Blood pressure was obtained from each participant with mercury sphygmomanometer.The levels of sVEGFR2 and SOD were measured by ELISA.Meanwhile, the levels of plasma glucose, glycosylated hemoglobin A1c ( GHbA1c) and lipid profile were detected.RESULTS:The levels of total cholesterol ( TC) and body mass index (BMI) were significantly higher in hypertensive group than those in control group (P<0.05).The levels of TC, low-density lipoprotein cholesterol ( LDL-C) , triglyceride ( TG) , BMI, waist circumference were significantly higher in hypertensive diabetic group than those in control group (P<0.05).The plasma levels of sVEGFR2 and SOD in both hypertensive diabetic group and hypertensive group were significantly decreased compared with control group ( P<0.05), while the mean plasma levels of sVEGFR2 and SOD in hypertensive diabetic group were significantly decreased compared to the hypertensive group ( P<0.05 ) .A significantly positive correlation between sVEGFR2 and SOD in the whole study population (P<0.05) was observed.CONCLUSION: The plasma level of sVEGFR2 is decreased in both hypertensive and hypertensive diabetic patients, and more significantly decreased in hypertensive diabetic patients.De-creased SOD level may be associated with to the reduction of sVEGFR2.

BACKGROUND: Fibrin glue has been demonstrated to function as a kind of biomaterial with high quality. It has been used in nerve tissue engineering and proved to be a kind of scaffold for some cells.OBJECTIVE: To explore the biocompatibility of fibrin glue and olfactory ensheathing cells (OECs).DESIGN, TIME AND SETTING: An in vitro control trial based on cytology was performed at the Institute of Neurobiology,Nantong University from August 2007 to February 2008.MATERIALS: Fibrin glue was made of fibrin and catalyst, and OECs derived from rats' olfactory bulb were normally primary-cultured.METHODS: OECs were divided into control (OECs clone spheres were cultured alone) and in fibrin glue (OECs clone spheres were cultured and combined with fibrin glue) groups. After 1 week of culture, the proliferation of OECs were observed by convert microscope and detected by S-100 immunofluorescence histochemical staining.MAIN OUTCOME M EASURES: OECs morphology, cell count, the area of the cell bodies and the perimeter of the cell were determined.RESULTS: OECs could survive, migrate in fibrin glue, and float in the fibrin glue in the lower layer. After 7 days of incubation, cell body exhibited fusiform or triangle, predominantly bipolar or bipolar. The number of the S-100 positive cells was more, and cell bodies were larger in fibrin glue group than control group (P < 0.05). However, there was no obvious difference between two groups in cell perimeter (P > 0.05).CONCLUSION: Fibrin glue has good biocompatibility with OECs, and OECs can survive and migrate in fibrin glue.