OBJECTIVE: To investigate the effects of apple polyphenols on pulmonary vascular remodeling in rats with pulmonary arterial hypertension and its mechanism. METHODS: Rats were randomly divided into 4 groups:control (Con) group, monocrotaline (MCT) group, apple polyphenol (APP) group,monocrotaline + apple polyphenol (MCT+APP) group. In Con group, rats received a subcutaneous injection of physical saline. In APP group, rats received intraperitoneal injection of 20 mg/kg APP, every other day. In MCT group, rats received a single subcutaneous injection of MCT(60 mg/kg). In MCT+APP group, rats received subcutaneous injection of 60 mg/kg MCT followed by an intraperitoneal injection of 20 mg/kg APP every other day. All the disposal lasted 3 weeks. Then the PAH-relevant indicators, such as mean pulmonary artery pressure(mPAP), pulmonary vascular resistance(PVR), right ventricular hypertrophy index (RVHI) ,wall thickness (WT%) and wall area (WA%) were tested. After that, the inflammatory pathway related indicators, such as interleukin1(IL-1),interleukin1(IL-6), tumor necrosis factor α(TNF-α), cyclooxygenase 2(COX-2) and myeloperoxidase(MPO) in pulmonary tissue and free intracellular Ca in pulmonary smooth muscle cell(PASMC), content of eNOS and NO in endothelial cells were determined. RESULTS: Compared with the control group, the levels of mPAP, PVR, RVHI, WA%, WT%, and IL-1, IL-6, TNF-α, COX-2, MPO in tissue and the expression of Ca in PASMC of MCT group were increased significantly, while the contents of eNOS and NO in endothelial cells were decreased significantly (P＜0.05). Compared with the MCT group, the apple polyphenol treatment could improve the above mentioned situation, and the COX-2 and Ca indicators of the apple polyphenol treatment group were decreased significantly (P＜0.05). CONCLUSION MCT can increase COX-2 expression and intracellular Ca in pulmonary artery smooth muscle cells, decrease the contents of eNOS and NO in endothelial cells, while apple polyphenols can significantly inhibit these effects.
Animals ; Calcium ; metabolism ; Cyclooxygenase 2 ; metabolism ; Cytokines ; metabolism ; Malus ; chemistry ; Monocrotaline ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Polyphenols ; pharmacology ; Pulmonary Artery ; drug effects ; pathology ; Random Allocation ; Rats ; Vascular Remodeling ; drug effects

Objective To investigate the promoter methylation of drug metabolism genes CYP1A1 and CYP1B1 in thyroid cancer and its relationship with clinical pathological characteristics. Method 201 cases of thyroid cancer and 23 cases of normal thyroid tissues were involved. Methylation-specific PCR ( MSP ) was performed to analyze promoter methylation of CYP1A1 and CYP1B1 genes in the above tissues to detect the frequency of methylation positive, compare the promoter methylation level of CYP1A1 and CYP1B1 genes in papillary thyroid carcinomas ( PTC) and the controls. Five thyroid cancer cell lines were treated with methyltransferase inhibitor 5-Aza-dC for 5 days, and real time PCR ( RT-PCR) was performed to evaluate the mRNA expression of CYP1A1 and CYP1B1 genes. Logistic regression was used to analyze the correlation between the aberrant methylation and the clinical features. Results Aberrant methylation status in promoter region of CYP1A1 and CYP1B1 genes were detected in all kinds of thyroid cancers. Compared with control tissues, the methylation in promoter regions of CYP1A1 in PTCs was significantly higher, while that in promoter regions of CYP1B1 was lower (P<0.05). In vitro, 5-Aza-dC treatment significantly increased the CYP1A1 gene mRNA expression for 6. 92 and 8. 30 times in K1 and C643 cell lines respectively and restored CYP1B1 gene mRNA expression for 7.62 times in K1 cell line. Compared with the controls, PTCs with methylation in promoter regions of CYP1B1 had decreased lymphatic metastasis rate. Conclusion The methylation in promoter regions of CYP1A1 and CYP1B1 gene may regulate their mRNA expressions. Aberrant methylation of the promoter region of CYP1B1 is associated with lymph node metastasis in PTC.

OBJECTIVETo explore the effects of the shock wave on the capacity of mesenchymal stem cells(MSCs) to proliferate and differentiate into osteoblasts.

METHODSMSCs were isolated from the bone marrow of healthy donors. The human bone marrow MSCs(BM-MSCs) were divided into 3 groups including blank control group,osteoinduced group and shock wave group. The MSCs in blank control group were cultured with common mediam; the MSCs in osteoinduced group were treated with osteogenic agents and cultured; the MSCs in shock wave group were cultured with common medium and stimulated by shock wave. The morphology of MSCs in each groups were observed by micoscopy; the CCK-8 was used to detect the proliferation ability of MSCs; the alkaline phosphatase staining and von Kossa staining were used to evaluale the differentiation potential of MSCs in each groups.

RESULTSThe results of CCK-8 revealed the shock wave could promote cell proliferation as compared with blank control group. The results of alkaline phosphatase and Von Kossa staining showed that the shock wave displayed a stronger ability to promote the human BMMSC differentiation into osteoblasts cells in comparison with the osteoinduced group. The blank control group was weakly positively stainined.

CONCLUSIONThe shock wave treatment can promote proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVETo explore the clinical value of detecting adenosine triphosphate (ATP) level in CD4T lymphocytes (Immuknow ATP) of patients on early stage after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe base-line ATP value in CD4T lymphocytes in cases of hematological malignancies and the ATP level in CD4T lymphocytes of acute leukemia patients before allo-HSCT were detected. Allo-HSCT recipients were devided into 3 groups with different level of immunereactivity according to ATP concentraiton in month 3 (day 90±5) after allo-HSCT. The clinical characteristics of patients in 3 groups were analyzed.

RESULTSThe mass concentration of Immuknow ATP in 15 cases of hematological malignancies before allo-HSCT ranged from 56.21-435.71 ng/ml, with a mean of 203.98±112.72 ng/ml. The ATP level in 46 cases after allo-HSCT ranged from 1.69-333.09 ng/ml, with a median of 41.96 ng/ml. Both 91.26 ng/ml (mean-SD) and 316.70 ng/ml (mean+SD) were used as cutoff, and 36 allo-HSCT recipients (78.3%) were assigned to low immunereactivity group, 8 recipients (17.4%) to middle group and 2 recipients (4.3%) to high group. The incidence of infection in low immunereactivity group was significantly higher than that in middle immunereactivity group (86.1% vs 50.0%)(P=0.022), and also significantly higher than that in high immunereactivity group (86.1% vs 0%)(P=0.002). There were no statistical differences in the incidences of severe infection among 3 groups. The incidence of grade II or higher acute graft versus host disease (aGVHD) in high immunereactivity group was superior to that in low immunereactivity group statistically (100% vs 13.9%)(P=0.002). Immune-mediated organ injury occurred more frequently in high immunereactivity group as compared with low and middle immunereactivity groups (100% vs 0% and vs 0%)(P=0.000; P=0.002). There were no significant differences in relapse rates of leukemia among 3 groups. The percentage of patients with increased trough blood concentration of cyclosporine A(CsA) was not significantly different among 3 groups (P=0.720).

CONCLUSIONDetection of ATP level in CD4T lymphocytes on early stage after allo-HSCT possesses clinical significance for predicting infection, severity at aGVHD and immune-mediated organ injury.

OBJECTIVEThough the rabbit is one of most widely used experimental animals for medical regenerative research, it remains difficult to culture mesenchymal stem cells (MSC) on a in large scale due to the extremely lower number and hematopoietic cell contamination. This study was aimed to establish a novel protocol to generate rabbit MSC by culturing bone marrow plugs instead of bone marrow cells so as to obtain a large amount of MSC with higher proliferation and self-renewal properties.

METHODSThe primary MSC were generated from collagenase digested bone marrow plugs and bone marrow cells, respectively. The surface antigen profile of MSC was analyzed with flow cytometry and the cells were induced to differentiate into osteoblasts and adipocytes. The proliferation capacity of MSC were assessed by CCK-8 method. To test their self-renewal property, the colony forming unit-fibroblast assay was performed. Moreover, the cell yields of passage 1, 2, 3 and 4 were calculated.

RESULTSThe bone marrow plug-derived MSC shared the typical fibroblast-like morphology same as bone marrow cells derived MSC. Moreover, the ratio of CD45 positive hematopoietic cells in bone marrow plug-derived MSCs was significantly lower than that of bone marrow cell-derived MSC. The results of multi-differentiation experiments showed that bone marrow-plug-derived MSC exhibited similar multi-potent property to their bone marrow counterparts. In addition, the results of CCK-8 and CFU-F assay demonstrated that bone marrow plug-derived MSC grew more robustly and more CFU-F were formed in the culture plates, which indicated that the cells possessed higher proliferation and self-renewal capacities. Promisingly, a larger amount of cells were harvested via using the new protocol.

CONCLUSIONThe purity and yields of the bone marrow plug-derived MSC are satisfactory compared with previous rabbit MSC isolation methods. The findings may be helpful for the research of regenerative medicine.

Adipocytes ; Animals ; Bone Marrow ; Bone Marrow Cells ; Cell Differentiation ; Cell Separation ; Colony-Forming Units Assay ; Mesenchymal Stromal Cells ; Osteoblasts ; Rabbits

OBJECTIVETo evaluate and summarize the possibility and experience of reconstructive vascular operation for kinking of internal carotid artery.

METHODSReconstructive vascular operation was performed on 5 patients with symptomatic kinking of internal carotid artery between July 2008 and June 2009. There were 2 male and 3 female patients, age ranged from 53 to 68 years (mean 62 years). Cutting the internal carotid artery at the bifurcation, mobilizing and stretching the internal carotid artery, then anastomosing the internal and common carotid artery. Two of them underwent endarterectomy simultaneously.

RESULTSFor the 5 patients, postoperative recovery went smoothly and symptoms were well improved. Except that mild high perfusion syndrome happened in 1 patient, no other obvious complications. Kinking of internal carotid artery had been stretched in the postoperative CT angiography. In the 5-16-month follow-up, no transient ischemic attack or cerebral infarction happened, and no restenosis appeared.

CONCLUSIONSReconstructive vascular operation is an effective surgical approach to kinking and coiling of the internal carotid artery. For some risk exists, all-round evaluation should be performed before operation, and operative indication should be strictly controlled.

Aged ; Carotid Artery, Internal ; Carotid Stenosis ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Vascular Surgical Procedures ; adverse effects ; methods

ObjectiveTo investigate the current status of Kaschin-Beck disease in Shandong province, and to provide a scientific basis for decision-making in controlling the disease. Methods According to the National Monitoring Program of Kaschin-Beck disease requirements, historical serious villages of Kaschin-Beck disease in Qingzhou of Shandong province were selected annually; children aged 7 to 16 were chosen to receive clinical examination and children aged 7 to 12 were taken X-ray examination. Clinical and X-ray diagnosis was carried out according to the Diagnostic Criteria of Kashin Beck Disease(GB 16003-1995). Results From 1996 to 2010, in 53 diseased villages, three thousand three hundred and eighteen school children aged 7 to 16 were clinically diagnosed, and child Kaschin-Beck disease of degree Ⅰ and above were not detected; three thousand and ninety-one school children aged 7 to 12 were examined by X-ray, forty cases were found positive, and the total positive rate was 1.29％(40/3091 ). The year with the highest positive rate was 2002, and the rate was 3.49％(13/372) ; the positive rate was 0 in 1996 and 2008. The difference of the X-ray positive rate between each year was statistically significant(x2 =31.54, P ＜ 0.01 ). ConclusionsChild Kashin-Beck disease in Qingzhou is basically under control.Since etiology of Kashin-Beck disease is still unclear, surveillance of the disease still needs to be strengthened.

OBJECTIVETo explore the affecting factors on coronary heart disease among people over 40 years of age in Guangxi area, China.

METHODSBaseline data was gathered through the Third National Blood Pressure Survey in 1991 in China. A total number of 11 818 adults over 40 years old had been studied in Guangxi province. Data of morbidity and mortality of coronary heart disease was obtained.

RESULTSCardiovascular events were related to systolic blood pressure, diastolic blood pressure, pulse pressure, smoking, BMI regardless of their myocardial infarct (MI) history. Cox regression analysis showed that the relative risk for cardiovascular events increased by 21 [95% confidence interval (CI): 9.06-48.44] times for those people having MI history. When pulse pressure, systolic blood pressure, diastolic blood pressure increased by every 10 mm Hg, the relative risk for cardiovascular events increased by 1.29 (95% CI: 1.11-1.49), 1.18 (95% CI: 1.02-1.22), 1.13 (95% CI: 1.05-1.28) respectively. There was 1.23 (95% Cl: 1.05-1.45) times higher in smoker than non-smoker on relative risk for cardiovascular events. When BMI increasing 1, the relative risk for cardiovascular events would increase 1.03 (95% CI: 1.01-1 .05) times.

CONCLUSIONHypertension, smoking, increase of BMI were the risk factors of coronary heart disease among people who were over 40 years of age.

Adult ; Body Mass Index ; China ; epidemiology ; Cohort Studies ; Confidence Intervals ; Coronary Disease ; epidemiology ; etiology ; Health Surveys ; Humans ; Hypertension ; complications ; Myocardial Infarction ; Proportional Hazards Models ; Risk Factors ; Smoking ; adverse effects

OBJECTIVETo detect the mutations of Krit-1 gene that cause familial cerebral cavernous malformation (CCM) in the Han ethnic origin.

METHODSThe subjects were hospitalized in the Department of Neurosurgery, Tiantan Hospital affiliated to Capital University of Medical Sciences. Two families (A and B) and 8 apparently sporadic individuals affected with CCM were screened for mutations of Krit-1 gene. Members of the family CCM have a wide range in age of onset with seizures, headaches and skin lesions. The gene was screened by PCR amplification of 16 exons and mutation was detected by direct sequencing.

RESULTSIn family A samples, analysis of the Krit-1 gene revealed a new point mutation in exon 14 [a heterozygous C to G transition at nucleotide 1 289 (counting from the start codon or nt 2 308 counting from the first nt of the mRNA, aligned according to Gene Bank AF388384)] which predicts the substitution of a premature termination codon for Serine at codon 430 (S430X), belonging a nonsense point mutation. No mutation was identified in one of family A members as well as in any of the sporadic individuals with the exception of a single nucleotide polymorphism.

CONCLUSIONSReport the first family in the Han with CCM having a novel mutation in the CCM1 gene on the continent of Asia. The newly identified mutation creates a premature termination codon and is predicted to produce a truncated Krev1 interaction-trapped 1 protein, KRIT1. This result allows efficient presymptomatic molecular diagnosis.

Adult ; Base Sequence ; Child ; Child, Preschool ; Female ; Hemangioma, Cavernous, Central Nervous System ; genetics ; pathology ; Humans ; KRIT1 Protein ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Molecular Sequence Data ; Mutation ; Proto-Oncogene Proteins ; genetics

In order to develop clinical diagnostic tools for rapid detection of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.
Antigens, Viral ; immunology ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; SARS Virus ; genetics ; immunology ; Yeasts ; genetics