of glucose and insulin are also marked after congee intake.

OBJECTIVETo investigate the expression of myeloid differentiation protein-2 (MD-2) in the human endothelial cells and its role in lipopolysaccharide (LPS)-induced NF-kappaB activation in endothelial cells.

METHODSIn vitro cultured human umbilical vein endothelial cells (HUVEC) were employed in the study. The expression of MD-2 mRNA and protein, and the effect of LPS on the expression of its mRNA and protein were assessed with RT-PCR and Western blotting. The role of MD-2 in LPS-induced NF-kappaB activation and IL-8 production were investigated with gene transfection of mutant MD-2 cDNA (0.5, 1.0, 2.0 microg), pEF-BOS vacant vector (2.0 microg) and MD-2 plasmid (2.0 microg) into HUVEC, respectively.

RESULTSThere was MD-2 mRNA and protein expression in HUVECs before LPS stimulation, and it could be obviously upregulated by LPS in time and dose-dependent manner (MD-2 protein absorbency was 25 196 +/- 1 723 without LPS stimulation, which was obviously lower than that stimulated with 0.01 mg/L LPS (58 817 +/- 3 241, P < 0.01) for 6 hours. Transfection of mutant MD-2 cDNA could remarkably inhibit LPS-induced NF-kappaB activation and IL-8 production in endothelial cells.

CONCLUSIONMD-2 might play an important role in the LPS-induced NF-kappaB activation in HUVECs.

Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; metabolism ; Humans ; Interleukin-6 ; genetics ; Interleukin-8 ; metabolism ; Lipopolysaccharides ; pharmacology ; NF-kappa B ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells.

METHODSThe designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method.

RESULTSThe proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation.

CONCLUSIONSThese new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.

Antigens, Surface ; analysis ; Base Sequence ; Biological Assay ; Cells, Cultured ; DNA ; analysis ; genetics ; Gene Expression Profiling ; methods ; Humans ; Lipopolysaccharide Receptors ; analysis ; Lymphocyte Antigen 96 ; Membrane Glycoproteins ; analysis ; Molecular Probe Techniques ; Molecular Sequence Data ; Monocytes ; metabolism ; RNA Probes ; analysis ; genetics ; Receptors, Cell Surface ; analysis ; Receptors, Immunologic ; analysis ; Ribonucleases ; Toll-Like Receptor 4 ; Toll-Like Receptors

BACKGROUNDThe decrease of surfactant protein (SP) secreted by the alveolar type II cell is one of the important causes of limiting air of pulmonary emphysema. However, the SP-A gene and protein changes in this disease are rarely studied. This study was undertaken to investigate alterations in SP-A gene activity and protein, and to explore their roles in the pathogenesis of emphysematous changes.

METHODSTwenty Wistar rats were divided randomly into a normal control group (n = 10) and a cigarette smoking (CS) + lipopolysaccharide (LPS) group (n = 10). Ultra-structural changes were observed under an electron microscope. The number of cells positive for SP-A was measured by immunohistochemistry. The mRNA expression and protein level of SP-A in the lung tissues were determined by quantitative polymerase chain reaction (qPCR) and Western blot separately. The protein level of SP-A in lavage fluid was determined by Western blot.

RESULTSThe number of cells positive for SP-A of the CS + LPS group (0.35 +/- 0.03) was lower than that of the blank control group (0.72 +/- 0.06, P < 0.05). The level of SP-A in the lung tissues of rats in the CS + LPS group (0.2765 +/- 0.0890) was lower than that in the blank control group (0.6875 +/- 0.1578, P < 0.05). The level of SP-A in the lavage fluid of rats in the CS + LPS group (0.8567 +/- 0.1458) was lower than that in the blank control group (1.3541 +/- 0.2475, P < 0.05). The lung tissues of rats in the CS + LPS group showed an approximate increase (0.4-fold) in SP-A mRNA levels relative to beta-actin mRNA (P < 0.05).

CONCLUSIONSThe changes of SP-A may be related to emphysematous changes in the lung. And cigarette smoke and LPS alter lung SP-A gene activity and protein homeostasis.

Animals ; Blotting, Western ; Emphysema ; metabolism ; pathology ; Homeostasis ; Immunohistochemistry ; Male ; Microscopy, Electron ; Polymerase Chain Reaction ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the clinicopathologic features and histogenesis of calcifying fibrous tumor (CFT).

METHODSThe clinical manifestations, histopathologic characteristics and immunophenotype were analyzed in 11 cases of CFT.

RESULTSThe male-to-female ratio was 5:6, with a mean age of 38 years and age range of 25 to 52 years. The sites of involvement included abdominopelvic cavity (n=6), soft tissue (n=4) and scrotum (n=1). Most patients presented with a gradually enlarging and painless mass. Nearly half of the cases were associated with other diseases or history of inflammation, trauma or surgical intervention. One third of the tumors represented incidental findings and showed no recurrence after resection. Imaging revealed a solitary solid soft tissue mass or multiple nodules with clear borders and associated high-density calcifications. Macroscopically, the tumors were well-circumscribed but non-encapsulated. They ranged from 0.5 to 20.0 cm in diameter and were tan-greyish, round to oval, lobulated or irregular and solid with rubbery consistency. The cut surface was whitish to tan-yellowish, gritty and showed scattered spotty yellowish discoloration corresponding to the foci of dystrophic calcifications. Histologically, CFT was composed of hyalinized fibrous tissue and thickened vessel walls with interspersed bland spindly fibroblastic cells, scattered psammomatous calcifications, dystrophic calcification and lymphoplasmacytic infiltration. In addition, focal cloak-like polymorph infiltration at the tumor periphery and entrapment of adipocytes and nerves were demonstrated in some cases. Foci resembling solitary fibrous tumor, fibromatosis, keloid or inflammatory myofibroblastic tumor were observed. Immunohistochemical study showed that the tumor cells were diffusely positive for vimentin and focally positive for CD34, factor VIII-related antigen and beta-catenin. The admixed plasma cells were notably IgG positive, with more than 50% being IgG4 positive.

CONCLUSIONSCFT has characteristic histopathologic manifestations and shows morphologic and immunohistochemical overlaps with known IgG4-related sclerosing diseases. It is possible that CFT may represent another example of IgG4-related diseases. It often runs a benign clinical course, with rare recurrence after surgical resection. Previous inflammation and trauma may be the precipitating factors of CFT.

Abdominal Neoplasms ; metabolism ; pathology ; surgery ; Adult ; Antigens, CD34 ; metabolism ; Calcinosis ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Genital Neoplasms, Male ; metabolism ; pathology ; surgery ; Humans ; Immunoglobulin G ; metabolism ; Incidental Findings ; Male ; Middle Aged ; Neoplasms, Fibrous Tissue ; metabolism ; pathology ; surgery ; Pelvic Neoplasms ; metabolism ; pathology ; surgery ; Retrospective Studies ; Scrotum ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; beta Catenin ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo explore VEGF siRNA's effect on the immature fetal retinal microvascular endothelial cells in vitro.

METHODThe fresh retinal micrangium was primarily cultured to obtain microvascular endothelial cells. CoCl2 was used to simulate oxygen-deficient conditions. siRNA directed against human VEGF was designed and chemically synthesized. There were 3 groups in our experiment: VEGF siRNA group, hypoxia control group, and negative siRNA control group. The fetal retinal micrangium vascular endothelial cells were transfected by using liposome. The expression levels of VEGF mRNA and protein were evaluated by RT-PCR and Western blotting 24, 48, 72 h after transfection, cell proliferation was evaluated by MTT method.

RESULTThe expression levels of VEGF mRNA decreased by 21.05%, 79.67%, and 90.48% 24 h, 48 h, and 72 h after transfection as compared to those in hypoxia control group, the expression level of VEGF protein had decreased by 14.58%, 66.97%, and 81.61% as compared to those in hypoxia control group. The siRNA could decrease cell proliferation under hypoxia too, the multiplication rate after 12, 24, 48, and 72 h decreased by 15.0%, 42.9%, 78.3% and 65.9%.

CONCLUSIONVEGF siRNA could down-regulate the expression of VEGF in immature fetal retinal microvascular endothelial cells and suppressed cell proliferation. Application of siRNA to inhibit expression of VEGF may be a hopeful way to prevent and cure ROP.

Cell Hypoxia ; Cell Line ; Endothelial Cells ; metabolism ; Humans ; Infant, Newborn ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Retina ; metabolism ; pathology ; Retinal Vessels ; cytology ; metabolism ; Retinopathy of Prematurity ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate the overall diet quality and diet model of labor workers in Shenzhen using Chinese Diet Balance Index (DBI).

METHODSIn May 2009, 14 canteens from Baoan, Longgang and Nanshan districts were selected by stratified random sampling and 60 workers were randomly selected from each canteen by using random number method. Diet measurements were carried out among the 840 labor workers. Diet quality was evaluated by using DBI scoring and evaluating system.

RESULTSThe median values of labor workers' food intakes of cereal and meat & poultry were 483.8 and 121.7 g/d, which were more than the recommended amounts of their intakes of Chinese residents (cereal: 250 - 400 g/d, meat & poultry: 50 -70 g/d). The median values of the labor workers' intakes of fruit, dairy and eggs were 37.3, 20.6 and 23.5 g/d,which were less than recommended amounts in fruits (200 - 400 g/d), dairy (300 g/d) and eggs (25 - 50 g/d). The DBI-HBS scores of males and females in Shenzhen migrant workers were 24.4 +/- 6.1 and 22.6 +/- 6.3, respectively with a statistically significant difference (t = 4.21, P < 0.01). DBI-HBS scores of < 20 age group, 20 - 29 age group, 30 - 39 age group and > or = 40 age group in labor workers were 12.7 +/- 5.9, 11.3 +/- 6.3, 12.8 +/- 6.4 and 11.2 +/- 5.6 respectively (F = 3.67, P = 0.01). There were 7 dietary patterns among labor workers in this survey. Nearly 8.2% (68/830) of them belonged to Pattern A. Pattern B and E were the main dietary patterns, which accounted for 37.3% (310/830) and 31.0% (257/830) of the total population.

CONCLUSIONDBI can describe and evaluate the overall dietary quality and the major problem of the dietary patterns in labor workers. It is necessary to strength nutritional education to increase the intake of fruits, milk and eggs to improve nutritional status in labor workers in Shenzhen.

Adolescent ; Adult ; Dairy Products ; Diet ; statistics & numerical data ; Diet Surveys ; Eggs ; Feeding Behavior ; Female ; Fruit ; Humans ; Male ; Meat ; Middle Aged ; Nutritional Status ; Young Adult

OBJECTIVETo investigate the gene mutations of CHRNB2 and CHRNA2 in Chinese population with autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE).

METHODSOne hundred and six Han nationality patients (74 sporadic and 32 familial) were recruited and studied. Mutational screening was performed by sequencing all the 6 coding exons of the CHRNB2 gene and exons 6 and 7 of the CHRNA2 gene including the donor and acceptor splice sites.

RESULTSThe results excluded the involvement of all known published mutations of the CHRNB2 and CHRNA2 genes. However, a novel synonymous mutation c.483C>T (H161H) and a single nucleotide polymorphism (c.1407C>G) of CHRNB2 gene were detected in two ADNFLE sporadic patients respectively. The nucleotide variation H161H was heterozygous and absent in 200 healthy control samples. The mutation was also found in the proband's unaffected mother.

CONCLUSIONOur study suggests that the mutations of CHRNB2 and CHRNA2 genes may be rare in Chinese ADNFLE population. The novel synonymous mutation of H161H has not been reported previously and its impact on the pathogenesis of ADNFLE needs to be further studied.

Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; Epilepsy, Frontal Lobe ; genetics ; Female ; Genes, Dominant ; Humans ; Male ; Mutation ; Receptors, Nicotinic ; genetics

Objective To explore the effect of reinforcement education on peritoneal dialysis-related peritonitis' prevention. Methods 991 patients who initiated continuous ambulatory peritoneal dialysis (CAPD) from January 2006 to December 2009 were recruited. The including standard was the duration on PD ＞ 3 months. The changing of peritoneal dialysis-related peritonitis' causes was analyzed regularly. The reinforcement education about peritoneal dialysis-related peritonitis' prevention was carried out by using training and retraining, follow-up, telephone call, leaflet,etc. Results Patients' mean age was (49.2 ± 16.1 ) ,male 56.5%,female 43.5% , Peritoneal dialysis duration 3.0 ～ 170.9 months (median 20. 1 months ), the primary renal disease was glomerulonephritis ( 59.2% ). After reinforcement education, the incidence of peritonitis was prolonging gradually ( from 57.2 patient months to 61.3 patient months). Comparing the causes of peritonitis in 2006, the proportion of dialysate connection' problems was significantly lower in 2009 ( x2 = 8. 229, P = 0. 004 ),the proportion of exit-site infection' problems was low continuously in 2009 ( ＜5% ), but the proportion of gastrointestinal problems was significantly increased ( x2 = 9. 720, P = 0. 002 ). Conclusions Reinforcement education could reduce the incidence of peritoneal dialysis-related peritonitis, especially reduce the incidence of peritonitis due to dialysate connection' problems. The prevention education about peritonitis caused by gastrointestinal problems need to be investigated further.

OBJECTIVETo investigate the prevalence, current treatment, and clinical characteristics of asthma, as well as the risk factors for this disease, among children aged 0-14 years in 2010 in urban Zhongshan, China.

METHODSA total of 10 336 children aged 0-14 years were selected from urban Zhongshan by cluster random sampling. The Third National Childhood Asthma Epidemiological Questionnaire 2010 was used to analyze the prevalence, current treatment, and clinical characteristics of childhood asthma, as well as the risk factors for this disease.

RESULTSAsthma was diagnosed in 179 cases (1.73%). The prevalence of asthma in male children was significantly higher than that in female children (2.25% vs 1.16%; P<0.01). Of the 179 patients, severe attacks were common in 104 cases (58.1%), 110 cases (61.5%) had slow onset, 102 cases (57.0%) had gradually relieved conditions, 61 cases (34.1%) suffered from asthma during seasonal transition, and 150 cases (83.8%) developed asthma due to respiratory tract infection. Among all asthmatic children, 71.5% had been treated with inhaled corticosteroids, and 71.5% had been treated with bronchodilator. The multivariate logistic regression analysis showed that a history of penicillin allergy, a family history of allergy, food allergy, eczema, allergic rhinitis, cesarean delivery, family mould, and perinatal passive smoking were independent risk factors for childhood asthma.

CONCLUSIONSThe prevalence of childhood asthma in urban Zhongshan is on a high level, and is associated with gender. The treatment of asthma has been standardized, but still needs further improvement. The onset of asthma attack is influenced by various factors.

Adolescent ; Asthma ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Risk Factors ; Seasons ; Time Factors