BACKGROUND:Our previous studies have shown that mesenchymal stem cells play an immunomodulatory role in Th17 cells, but the mechanism of action remains to be elucidated, therefore, to explore the role of Tol-like receptor 1 in which wil provide possible experimental basis for the future potential of celltherapy strategies.
OBJECTIVE:To investigate the role of Tol-like receptor 1 in the immunoregulatory function of mesenchymal stem cells on Th17 cells.
METHODS:Separation of adherent bone marrow-derived human embryonic sources of mesenchymal stem cells, immunomagnetic separation of normal CD4+T cells. CD4+T cells were cultured alone or in combination with mesenchymal stem cells co-cultured 4d. Real-time quantitative polymerase chain reaction assay interleukin-17, Tol-like receptor 1 expression levels related genes;number of Th17 cells by flow cytometry. Bone marrow mesenchymal stem cells from human embryos were separated using adherence method, and co-cultured with human CD4+T cells from healthy donors using immunomagnetic separation method for 4 days. The expression of interleukin-17 and Tol-like receptor 1 mRNA was detected by real-time PCR, and the number of Th17 cells was observed by flow cytometry.
RESULTS AND CONCLUSION:Tol-like receptor 1 mRNA was expressed in both CD4+T cells and mesenchymal stem cells. Level of interleukin-17 mRNA was significantly higher in mesenchymal stem cells co-cultured with CD4+T cells than in CD4+T cells cultured alone (relative value 3.59±0.11 vs. 1.14±0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 1 mRNA was detected in mesenchymal stem cells co-cultured with CD4+T cells compared with CD4+T cells cultured alone (relative value 6.07±1.79 vs. 1.53±0.63, P<0.01). Furthermore, Flow cytometry analysis showed that the percentage of Th17 cells in the mesenchymal stem cells co-cultured with CD4+T cells was significantly higher than that in CD4+T cells cultured alone, (4.53±1.27)%vs. (2.39±0.80)%(P<0.01). Tol-like receptor 1 might be involved in the immunoregulatory regulation of Th17 cells by mesenchymal stem cells.

BACKGROUND:Previous studies have found that embryonic bone marrow mesenchymal stem cel s can promote human Th17 cel proliferation, but the inherent regulatory mechanisms stil need to be elucidated. OBJECTIVE:To investigate the role of Tol-like receptor 3 in the immunoregulation of Th17 cel s by mesenchymal stem cel s.
METHODS:Human CD4+T cel s from healthy donors were isolated by immunomagnetic bead method, and then cultured alone or co-cultured with embryonic bone marrow mesenchymal stem cel s for 4 days. The mRNA expression level of interleukin-17, Tol-like receptor 3 and MyD88 was detected by real-time PCR.
RESULTS AND CONCLUSION:Compared with CD4+T cel cultured alone group, the mRNA level of interleukin-17 was significantly higher in the co-culture group (3.59±0.11 vs. 1.14± 0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 3 mRNA was detected in the co-culture group compared with the CD4+T cel cultured alone group (3.10±1.60 vs. 0.94± 0.01, P<0.05). Furthermore, MyD88 in the co-culture group was significantly higher than that in CD4+T cel cultured alone group (2.29±0.05 vs. 1.85±0.31, P<0.01). Tol-like receptor 3 may be involved in the immunoregulation of Th17 cel s by embryonic bone marrow mesenchymal stem cel s, which provides experimental evidence for potential cel therapeutic strategy.

Objective To investigate the change of Connexin43 expression in rat heart as age advancing. Methods The expressions of Connexin43 in different age of rat heart were determined by immunohistochemistry. Results 1\^ The expression of Connexin43 was only seen in ventricular myocardium. 2\^ The Connexin43 immunolabelling of infant rat heart lie much more within the transversely orientated intercalated disk, besides, a fewer have a punclate distribution over the entire surface of the ventricular myocytes, and Connexin43 distributed mainly on the myocyte surface of the parpillary muscles. Connexin43 immunolabelling of young and old rat heart are typically confined to the site of intercalated disk.Conclusion The expressions of Connexin43 are different in different age and different parts of rat heart.

Objective To research the spatiotemporal pattern of N-cadherin expression during development of rat cardiomyocytes and age-related changes of N-cadherin expression. Methods Using immunohistochemical method,myocardial N-cadherins distribution was investigated in fetal rat and postnatal development(1 postnatal day to old rat),and quantitied by HIPAS-1000 computerized image analytical instrument. Results The expression of N-cadherin was located in myocardium of atrial and ventricule and septum interventriculare and papillary muscles.The N-cadherin immunolabeling was found in myocyte membranes and within cytoplasm in fetal rat heart.From neonatal to infant rat,the pattern of N-cadherin immunolabeling changed progressively,from being dispersed over the entire cell surface as in the fetal to the transverse terminals of the myocytes,toward the distribution within the intercalated disk.From young to old rat heart,the typical N-cadherin was located in transversely orientated intercalated disk.The percentage of N-cadherin immune postive area in rat ventricular myocardium showed a progressively changement with age.Conclusion The present paper demonstrated that the N-cadherin expression exists progressively with ages and the changing pattern of N-cadherin is closely related with development of the intercalated disk in rat myocardium.The mechanical coupling provided by adherens junctions is essential for the stable cell-cell contact.

Objective To investigate age-related changes of type Ⅰ and Ⅲ collagen in rat heart. Methods Twenty-one male Wistar rats were divided randomly into infanct,young and old group.Types Ⅰ and Ⅲ collagen expresson of different age hearts were studied by SP immunohistochemical methods. Results 1.Type Ⅰ and Ⅲ collagen all constitute thick fiber and fibril.The fibrils encircled every cardiac muscle and formed collagen net each other.The thick fibers being plaque were located among cardiac muscle groups.The content of type Ⅰ collagen was more than that of type Ⅲ.2.There were distinct expression difference of type Ⅰ and Ⅲ collagen in different aging rat hearts.The collagens distributed densely and evenly in infancy rat heart,and loosely and uniformly in young rat heart,and the contents were increased distinctly with heterogeneous distribution in old rat heart.The cardiac collagen was growing from infanct to old rat,and rapid progress of cardiac collagen was seen in young rat. 3.The content of collagen in right ventricle was more than that of left ventricle of infanct heart.The collagen in all parts of the heart is not of difference both in yound and old rat. Conclusion The contents of type Ⅰ collagen is more than that of type Ⅲ collagen.The two types collagen are increasing with growing.;

Objective To investigate the differences of connexin 43(Cx43)expression between adult and infant heart. Methods By using immunohistochemistry to observe the expression of Cx43. Results 1.The expression of Cx43 had a punclate distribution in cytoplasm and over the entire surface of the cardiocyte,and a few located at intercalated disk of atrial and ventricular myocardium in the infant heart.2.Cx43 positive granules distributed irregularly in cell side surface and cytoplasm as well as intercalated disk in adult atrium.Cx43 immunolabelling of adult ventricular myocardium was typically confined to the site of intercalated disk.3.The results of image analyzer showed that the amount of connexin43 expression was lower in the atrium than that of the ventricle in infant heart and atrium bigger than ventricle in adult heart.The expression of Cx43 was less in adult heart than that of infant heart.Conclusion The expression of Cx43 was mainly over the entire surface of the myocardium in infant heart.There were expression differences of Cx43 in human ventricular and atrial myocytes.The amount of Cx43 expression was higher in ventricle than that of atrium in infant heart and atrium bigger than ventricle in adult heart.It was less in adult heart than that of infant.It showed that the expression of Cx43 in human heart existed a developmental differences.

Objective To investigate expressional pattern of the Connexin 43 in atrial myocytes of ventricular septal defect children. Methods The expressional role of the Connexin 43 in atrial myocytes of ventricular septal defect children were observed and determinated by immunohistochemistry and digital image analysis technology. Results 1.The Connexin 43 granules of 1-3 year-old-group normal children were mainly distributed on cell side surface,which took on beening spot-,belt-or chain-shaped,but seldom at intercalated disks. The Connexin 43 of 7-16 year-old-group normal children were abundantly distributed on cell side surface and intercalated disks. 2.The Connexin 43 of 1-3 year-old-group ventricular septal defect children were not noly distributed on cell side surface but could be detected at intercalated disks and cytoplasm,a few granules were irregular in arrangement.The Connexin 43 of 7-16 year-old-group ventricular septal defect children were distributed on cell side surface and in cytoplasm but seldom existed at intercalated disks,and arranged in irregular order. 3.The experimental results showed insignificant difference in the expressional amount of the Connexin 43 in atrial myocytes between the ventricular septal defect children and normal children. Conclusion The Cx 43 distributional pattern in atrial myocytes of ventricular septal defect children was changed,but its expressional amount was indistinct,which suggested that ventricular septal defect affected only the Cx43 expressional pattern.

BACKGROUND: Type Ⅰ and type Ⅲ collagen, the main components of cardiac stroma,have supporting,protective and restrictive effects on myocardial cells. They are essential for the heart to maintain its normal shape and function. OBJECTIVE:To explore the effects of types Ⅰ and Ⅲ collagen on cardiac function by observing their changes in atrium myocardium of children with ventricular septal defect (VSD). DESIGN: A case analysis. SETTTING: Department of Cytobiology, Xinxiang Medical College. PARTICIPANTS: Six children who had died from accidents were selected from the Pediatric Department of the First Affiliated Hospital,Xinxiang Medical College, between January and August 2003. Informed consent was obtained form their relatives. They were 4 males and 2 females,ranging from 1 to 15 years in age. Congenital heart diseases were excluded by naked-eye anatomical examination,and right atrium tissues were collected for the study. Meanwhile, 21 children with VSD, 12 males and 9 females aged 1-17 years,were recruited from the Department of Cardiac-thoracic Surgery of the same hospital. METHODS:A small amount of fresh myocardium was cut from the right atrium at the edge of incision. Peroxidase-labeled strepto-avidin immunohistochemical method was adopted to detect the expression of types Ⅰ and Ⅲ collagen protein in atrium myocardium, which was analyzed using image analysis to calculate the area composition ratio of type Ⅰ and type Ⅲ collagen in atrium myocardium(representing the relative area percentage of collagen protein) and area percentage (representingthe expression percentage of collagen protein). MAIN OUTCOME MEASURES:Expression of typeⅠand type Ⅲ collagen protein in atrium myocardium. RESULTS: Six normal controls and 21 patients with VSD entered the final statistical analysis. ① Type Ⅰcollagen of normal atrium was strip-like fibers with different diameters connected with each other,whereas type Ⅲ collagen was scattered in spots and pieces. ② The distribution of type Ⅰ and Ⅲ collagen of atrium in VSD patients was mostly similar to that of the normal controls; only part of them displayed extreme rearrangement, that is,type Ⅰcollagen presenting large-speckle increment,breakage or disappearance, while type Ⅲ collagen increased in stripes and bundles. ③ The area percentage and area composition ratio of type Ⅰ and type Ⅲ collagen:type Ⅰ collagen expressed more than type Ⅲ collagen in normal myocardium [area percentage: (48.82±12.35)％ vs (40.02±13.53)％, t=2.173, P ＜ 0.05;area composition: (15.87±6.03) μm2 vs (13.62 6.94) μm2, t=2.221, P ＜ 0.05].Likewise, type Ⅰ collagen expressed more highly than type Ⅲ collagen in VSD myocardium [area percentage: (55.37±10.42)％ vs (50.27±14.36) ％,t=2.173, P ＜ 0.05; area composition: (24.93±9.62) μm2 vs (19.22 12.03) μm2,t=2.221, P＜ 0.05]. The content of both type Ⅰ and type Ⅲ collagen in VSD children was obviously higher than that in normal children(t=2.153-2.234,P ＜ 0.05). CONCLUSION: In children with VSD, part of type Ⅰ and type Ⅲ collagen is found rearranged and increased in atrium, which may result from the interstitial compensation due to cardiac dysfunction.

Objective To explore the CT and MRI findings of Kimura disease in different location.Methods The CT and MRI obtained in 19 patients with histologically proved Kimura disease were reviewed retrospectively.Results Of these 19 patients with Kimura disease, the head and neck was involved in 16 patients, other locations less commonly involved were the lung(n=1),chest wall(n=1),liver and spleen(n=1).Kimura disease of head and neck,the density abnormal appeared as nodular,mass or diffuse on CT imaging, the CT value of lesion was similar to muscle, MRI manifested as a slightly hypo-or isointense lesion on T1WI and hyperintense on T2WI,most of the lesions demonstrated moderate or marked enhancement on postcontrast CT and MR images.Kimura disease of lung and chest wall,lesion appeared as a solid mass on imaging, there was moderate enhancement in the lung lesion and rim enhancement in the chest wall lesion.Kimura disease of liver and spleen: lesion showed nodular, mild and gradual upward enhancement.Conclusion Imaging findings of Kimura disease are not characteristic, it is very essential to know the lesion combined with the laboratory and pathologic features.

OBJECTIVE: To analyze the association between genetic polymorphisms of DNA repair genes of XPD (751 Lys/Gln), XPC (PAT)and susceptibility to laryngeal carcinoma. To explore the effect between DNA repair genes of XPD (751 Lys/Gln), XPC (PAT) and carcinogenesis of LSCC(laryngeal squamous cell carcinoma). METHOD: A case-control study was conducted involving 233 LSCC patients and 102 healthy controls to investigate the association between polymorphisms of XPD(751 Lys/Gln), XPC (PAT) and LSCC. All blood samples of the Han people from the Guang Dong Zone was analysze with methods of PCR, PCR-RFLP, ASA and the technique of checking DNA sequencing with sequenator. We explored the association between polymorphisms and the clinical pathologic characteristic of LSCC. The data was compute with SPSS13.0. Odds Ratios (ORs) with 95% CI for relevancy intensity were calculated using binary logistic regression analysis. REULT: There is no difference of the frequency of XPC-PAT and XPD (751 Lys/Gln) genotype between in LSCC and in healthy contradistinguish (P > 0.05). CONCLUSION There may be no association between the susceptibility to laryngeal carcinoma and the genotype of XPC-PAT and XPD (751 Lys/Gln).
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; DNA Repair ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Genotype ; Humans ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Xeroderma Pigmentosum Group D Protein ; genetics