Objective To investigate the circulatory levels of hormones and cytokines in the patients with the chronic heart failure(CHF)and the effect of naftopidil.Methods The levels of tumor necrosis factor-?(TNF-?),interleukin-1(IL-1),interleukin-6(IL-6),endothelin-1(ET-1),adrenomedullin(Adm),neuropeptide Y(NPY),calcitonin gene-related peptide(CGRP),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP)and C-type natriuretic peptide(CNP),and left ventricular fraction(LVEF)of 47 patients with CHF(CHF group)and 24 healthy subjects(control group)were measured by radioimmunoassay or echocardiography respectively.The patients were randomized to receive naftopidil(50mg Bid)in addition to routine therapy(B group,n=23)or routine therapy only(A group,n=24)for one-month.Results (1)The levels of TNF-?,IL-1,IL-6,ET-1,Adm,NPY,ANP,BNP and CNP of the CHF group were higher(P 0.05)in the levels of IL-1,IL-6,Adm,CNP and LVEF.(4)There were significantly inverse correlations between LVEF and TNF-?,ET-1,IL-6 or BNP(P

Objective To identify differently expressed genes and pathways between Kashin-Beck disease (KBD) cartilage and healthy cartilage,and to explore the mechanism of articular cartilage lesions of KBD.Methods Cartilage specimens were collected from 9 patients with KBD and 9 healthy controls.Total RNA was extracted from cartilage specimens,and transcribed into cDNA.KBD and control groups were labeled by Cy3 and Cy5,respectively.Agilent genome-wide microarray was applied to compare the expression profile of KBD cartilage and healthy cartilage.The microarray data was analyzed by single gene and pathway expression analysis to identify differently expressed genes and pathways between KBD and healthy controls.Results ①Tweenty nine genes were significantly up-regulated in KBD group (averaged ratio =6.68 + 1.98,P ＜ 0.05),mainly involved in apoptosis,metabolism,extracellular matrix,cytoskeleton and cell movement.Additionally,extracellular matrix-related FBLN1 gene was down-regulated in KBD group(ratio =0.14 + 0.06,P ＜ 0.05).②Five apoptosis and 6 hypoxia-related pathways presented higher expression levels in KBD compared to healthy controls(all P＜ 0.05).Conclusions We find significant expression differences of apoptosis and hypoxia-related genes and pathways between KBD cartilages and healthy cartilages,suggesting that hypoxia might contribute to chondrocytes apoptosis of KBD.Further studies may be needed to investigate the relationship between hypoxia and articular cartilage lesions of KBD.

Objective To compare the expression profile of mycotoxin-related environmental response genes (MERGs) in the articular cartilage of patients with Kashin-Beck disease (KBD) and healthy controls,and explore the relationship between MERG and KBD.Methods Articular cartilage specimens were collected from 9 healthy human subjects and 9 adult KBD patients.Agilent microarray was used to evaluate the expression levels of MERG in cartilage specimens,and the expression ratios of MERG between KBD and healthy controls were calculated.GSEA software was used to calculate the NES scores and P values of gene ontology(GO).Results ①T-2 toxin,deoxynivalenol,zearalenone,aflatoxin B1,fumonisin B1 and ochratoxin A related 15 MERGs presented expression differences between KBD and healthy controls(ratios ＞ 2.0 or ＜ 0.5).Thirteen MERGs were up-regulated in KBD,including BAX,BCL2,COL5A2,FER1L3,GSTT2,IGFBP2,IGFBP4,PDE8B,SOCS3,THBS1,TMSL8,VGLL3 and TUBB2A (ratio ＞ 2.0).Two MERGs,POSTN and FABP4,were down-regulated in KBD (ratio ＜ 0.5).The 15 MERGs were involved in various biological processes; such as collage synthesis,apoptosis,metabolism,growth & development and so on.②Mycotoxin related 4 apoptosis GOs and 5 growth & development related GOs were up-regulated in KBD compared to healthy controls(NES ＞ 0),including ANTI_APOPTOSIS,REGULATION_OF_PROGRAMMED_CELL_DEATH,APOPTOSIS_GO,REGULATION_OF_APOPTOSIS,ORGAN_MORPHOGENESIS,ANATOMICAL_STRUCTURE_DEVELOPMENT,ORGAN_DEVELOPMENT,SYSTEM_DEVELOPMENT and REGULATION OF DEVELOPMENTAL_PROCESS (NES ＞ 0 and P ＜ 0.05).Conclusions There are multiple mycotoxins related environmental response genes presenting significant expression difference between KBD cartilage and normal cartilage.Mycotoxin can affect the expression of MERGs in KBD articular cartilage,which might lead to dysfunction of chondrocytes,and articular cartilage lesions.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

To introduce a method of classification with high precision-the artificial neural network (ANN),and to compare the results using logistic method. Using data from 1070 landless peasants'mental health survey,the artificial neural network models and logistic regression model were built and compared on their advantages and disadvantages of the two models.The prediction accuracy for artificial neural network was 94.229% and for logistic regression it was 51.028%. ANN appeared to have had good ability on generalization. ANN displayed advantages when conditions of classical statistical techniques could not be met or the predictive effect appeared to be unsatisfactory. Hence, ANN would make a better facture of its application in medical researche.

BACKGROUNDGlobally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.

METHODSAdult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.

RESULTSIn cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).

CONCLUSIONSA method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.

Adult ; Cell Separation ; methods ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Humans ; Islets of Langerhans ; physiology ; Islets of Langerhans Transplantation ; Male ; Sertoli Cells ; cytology ; physiology

OBJECTIVETo establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.

METHODSCD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.

RESULTSAfter 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).

CONCLUSIONThe method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Dendritic Cells ; cytology ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Proto-Oncogene Proteins c-kit

Background Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered ‘nurse cells'in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro. Methods Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro. Results In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P>0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P<0.01). Conclusions A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.

Objective To summarize the clinical features and outcomes of paraduodenal pancreatitis (PP). Methods Five clinically or pathologically diagnosed PP patients in Peking Union Medical College Hospital and 31 other PP cases reported in Chinese literature since 1988 were retrospectively analysed. Results Most PP patients were young or middle-aged males with a history of alcohol abuse. The clinical symptoms included upper abdominal pain,vomiting,weight loss,and fluctuating jaundice. Serum pancreatic enzymes were normal or elevated. Radiological features in most cases included thickening of the duodenal wall and duodenal stenosis (88.9%,32/36),cysts in the duodenal wall and groove area (47.2%,17/36),dilated bile duct (36.1%,13/36),and dilated pancreatic duct (16.7%,6/36). The main pathological finding was chronic pancreatitis,which could be accompanied by local acute inflammation,which was limited in the groove-duodenal area in most cases. The disease can be well controlled by conservative treatment,although surgery was needed in a small number of cases. Conclusion sPP typically occurs in young or middle-aged males. Radiological examination is valuable for diagnosis. Conservative treatment is the mainstream treatment in most patients.