Objective To study the repair function of united endothelial progenitor cells (EPC)transplantation on injured liver endothelium by bone marrow transplantation (BMT) conditioning.Methods C57BL/6 mice were divided into four groups randomly: normal control group, without any treatment; irradiation alone group, administered a total body irradiation(TBI) pretreatment, without BMT; (3) BMT alone group: C57BL/6 mice were infused with bone marrow mononuclearcells (MNC) 5 × 106/only through caudal vein not more than 4 h after the same TBI pretreatment as the irradiation alone group; united transplantation group: receiving the same way as the BMT alone group, but C57BL/6 mice were infused with EPC 5 × 105/only at the same time. Two, 4, 7, 14, and 21 days after the TBI, the changes of the liver weight were observed regularly. The histopathological examination of liver was done at the 4th, 7th, 14th, and 21st day after the TBI. Results In irradiation alone group, BMT alone group and united transplantation group the liver weight began to increase significantly on the day 2 and peaked at 14th day after the TBI, and the peaks were respectively (1.65±0. 15) times (P＜0. 05), (1.61 ±0.06) times (P＜0.05), and (1.11 ±0.40)times (P＜0. 05) of those in normal control group. At the day 14, the liver weight in irradiation alone group, BMT alone group and united transplantation group began to decrease, and on the day 21 the liver weight in united transplantation group had been completely restored to normal level, however the liver weight in irradiation alone group and BMT alone group were still significantly heavier than that in normal control group (P＜0. 05). Liver histopathological examination revealed that there were obvious sinusoidal endothelial cells (SEC) injury, hepatocyte edema and severe inflammatory cell infiltration in irradiation alone group, and on the day 7 the hepatocyte edema and necrosis were significantly worse than before, and almost no alive SEC were found. On the day 14 the injury of SEC in BMT alone group was lighter than before, but on the day 21 the injury had not returned to normal. On the day 7 the injury of SEC, hepatocyte edema and necrosis were alleviated in united transplantation group as compared with irradiation alone group and BMT alone group, and on the day 14 the injury had returned to normal basically. Conclusion The transplantation conditioning could damage recipient liver endothelium and the injury would persist, and united EPC infusion could repair the injured SEC following BMT.

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

BACKGROUNDThe p22phox is a critical component of the superoxide-generating vascular nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Several polymorphisms in p22phox gene are studied for their association with cardiovascular diseases. However, no publication is available to assess the relation of 549C > T polymorphism in p22phox gene to coronary artery disease (CAD) risk. This study was to investigate the effect of the p22phox gene 549C > T polymorphism on CAD risk.

METHODSHospital-based case-control study was conducted with 297 CAD patients and 343 healthy persons as the control group. Polymerase chain reaction and pyrosequencing using PSQ 96 MA Pyrosequencer (Biotage AB) were used to detect the polymorphisms. Multiple Logistic regression model was used to adjust the potential confounders and to estimate odds ratio (OR) with 95% confidence intervals (CIs).

RESULTSThe observed genotype frequencies of this polymorphism obeyed the Hardy-Weinberg equilibrium in both cases (P = 0.439) and controls (P = 0.668). The frequency of mutant genotypes (TT + CT) in cases (41.08%) was higher than that in controls (36.73%) with an OR = 1.20 (95%CI = 0.87-1.65). After the adjustment of the potential confounders, there was a significant association of the mutant genotypes with increased risk of CAD (OR = 1.57, 95%CI = 1.01-2.46, P = 0.047).

CONCLUSIONSThe mutant genotypes of the p22phox gene 549C > T polymorphism had a significant effect on the increased risk of CAD in this studied population.

Case-Control Studies ; Coronary Artery Disease ; etiology ; genetics ; Female ; Genotype ; Humans ; Logistic Models ; Male ; Middle Aged ; NADPH Oxidases ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo assess the association between G894T (Glu298Asp) mutation in exon 7 of the endothelial nitric oxide synthase gene and premature coronary heart disease (P-CHD).

METHODSHospital-based case-control study was conducted. Newly-diagnosed CHD patients were recruited as study subjects. 132 CHD patients diagnosed at/before age 55 for males and 65 for females were assigned to P-CHD case group with other 172 CHD patients as the control group. Polymerase chain reaction with Ban II restriction enzyme digestion was performed to detect the G894T mutation.

RESULTSG894T mutant genotypes in P-CHD group (TT, GT and GG frequencies were 6.06%, 20.45% and 73.48%, respectively) were significant higher than those in control group (TT, GT and GG frequencies were 1.74%, 11.63% and 86.63%, respectively) (P = 0.01). Mutant T allele frequency in P-CHD group was also significantly higher than that in control group (16.29% versus 7.56%, P = 0.001, OR = 2.38, 95% CI: 1.38 - 4.16). Stepwise multiple logistic regression analysis at 0.05 significant level with sex, smoking, alcohol drinking, and overweight covariates indicated that G894T mutation also having significant effect on P-CHD (P = 0.01, OR = 2.25, 95% CI: 1.19 - 4.26).

CONCLUSIONThis study suggested that G894T mutation in endothelial nitric oxide synthase gene might serve as a major risk factor to the pathogenesis of P-CHD in this study population.

Age Factors ; Case-Control Studies ; Coronary Disease ; enzymology ; genetics ; Exons ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase Type III ; genetics ; Point Mutation ; Risk Factors

Amputation ; Chronic Limb-Threatening Ischemia ; Fibrinolytic Agents/therapeutic use* ; Humans ; Limb Salvage ; Peripheral Arterial Disease ; Risk Factors ; Singapore ; Treatment Outcome

Objective To investigate the supply of iodized salt in non-excessive iodine counties and iodine-free salt in excessive iodine counties at household level in Hebei province so as to provide a basis for the prevention and control of iodine deficiency disorders(IDD). Methods According to the national project of IDD surveillance,the county was taken as the elementary sampling unit. The towns and villages were selected by systematic and random sampling in every county and households were chosen by random sampling to collect their edible salt in Hebei province in 2008. The salt iodine content in non-and excessive iodine regions was detected by direct titrition method and semiquantitative method respectively. Results all 48 448 salt samples were collected from 167 non-excessive iodine counties. Weighed by the population of counties,the rate of non-iodized salt was 4.73%. Iodized salt accounted for 95.27%,out of which,96.13% were qualified and the consuming rate of qualified iodized salt was 91.96%. Eighty point eighty three percent(135/167) of the counties covered by iodized salt above 95%,92.81% (155/167) passing rate of iodized salt above 90% and 82.04 (137/167) consuming rate of qualified iodized salt. All 1466 salt samples were collected in 5 counties with excessive water iodine content and the coverage rate of iodine-free salt was 93.25%(1367/1466). Conclusions In a nutshell,the national targets for preliminary elimination of IDD have been achieved in regions of non-excessive iodine of Hebei province. Nevertheless,the coverage rate of iodized salt and qualified iodize salt rate in some counties are still below the national standard. Therefore the prevention and control of IDD need to be strengthened. The supply of iodized salt in excessive iodine regions should be timely stopped.

Objective To understand the status of drinking-water-borne endemic fluorosis and the effect of preventive measure in Hebei province, so as to provide a basis to prevent and cure the disease. Methods Thirtyeight affected counties(cities, districts) with drinking-water-borne endemic fluorosis were sampled by random sampling in Hebei in 2009. All affected villages in every county were divided into mild, moderate and severe endemic fluorosis areas and a village was randomly selected from each category of the area to carry out the monitoring of endemic fluorosis. Dental fluorosis of children aged 8 - 12 were examined and 6 copies of urine samples were randomly collected in each age group in the above-mentioned villages. Clinical skeletal fluorosis was diagnosed among adults aged 16 and over and 20 copies of urine samples were tested for fluorosis in every village.Results A total of 112 affected villages were investigated, among which the drinking water quality of 66 villages were improved and 46 villages were not improved. A total of 236 copies of water samples from the 66 villages were measured and the fluoride content ranged from 0.1 to 4.3 mg/L, among which 20 copies of water samples exceeded the fluorine standard of 1.2 mg/L, accounting for 33.3%. A total of 230 copies of water samples were collected in the 46 villages and the fluoride content ranged from 0.2 to 4.6 mg/L, among which 76.1% (35/46) of the water samples exceeded the fluorine standard of 1.2 mg/L. A total of 5169 children aged 8 - 12 were examined of dental fluorosis, the dental fluorosis rate was 36.43%(1883/5169) and the dental fluorosis index was 0.81. A sum of 71 497 adults aged over 16 years were examined, and the rate of skeletal fluorosis was 4.81%(3438/71 497), moderate or severe clinical detection rate of skeletal fluorosis was 1.56%( 1114/71 497). A total of 2876 copies of children urine samples and 2021 copies of adult urine samples were tested and the geometric mean of fluoride content was 2.30,3.32 mg/L, respectively. Conclusions The prevalence of dental fluorosis of children in the areas with improved water is less than 30% and the rate of dental fluorosis and skeletal fluorosis decline gradually with time.The rate of dental fluorosis and skeletal fluorosis increases with the increase of water fluoride in the water quality not improved areas. The endemic fluorosis is still comparatively serious in Hebei. The progress of improving water quality in the areas with endemic fluorosis should be accelerated and the acceptability of improved water should be enhanced.

OBJECTIVETo analyze the association between G894T (Glu298Asp) mutation at exon 7 in the endothelial nitric oxide synthase gene and essential hypertension.

METHODSOne hundred and sixteen essential hypertensives without taking hypertensive medication and 136 normotensives screened from health workers in a steel factory were selected as subjects in this study. Polymerase chain reaction (PCR) and Ban II restriction enzyme digestion were performed to detect the G894T mutation.

RESULTSG894T mutation was significantly associated with essential hypertension. The T allele frequency in essential hypertensive group was significantly higher than that in normotensive group (16.0% versus 8.8%, P = 0.019, OR = 1.96, 95% CI: 1.14 - 3.37). The levels of systolic blood pressure and diastolic blood pressure in the G894T mutant genotypes were all significantly elevated in hypertensive, normotensive, and the total subjects (P < 0.05). After adjusting factors as age, sex, smoking, alcohol drinking, body mass index, triglyceride, serum total cholesterol, serum high density lipoprotein cholesterol by analysis of multiple covariance, significant positive effect of the G894T mutant genotypes on blood pressure in the total subjects (P < 0.01) was noticed.

CONCLUSIONThis study suggested that the G894T mutation in the endothelial nitric oxide synthase gene might serve as a major risk factor of essential hypertension in this study population.

Adult ; Alleles ; Blood Pressure ; genetics ; China ; epidemiology ; Exons ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Hypertension ; enzymology ; genetics ; Male ; Middle Aged ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type III ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Risk Factors

OBJECTIVETo explore the relationship between the 7th exon G894T mutation of endothelial nitric oxide synthase (eNOS) gene and overweight in patients with essential hypertension.

METHODSTotally, 116 patients with essential hypertension taking no medications and 136 normotensives were selected from a steel workers as study subjects. Polymerase chain reaction and restriction fragment length polymorphism were performed to detect mutation of the 7th exon G894T. Additive model was used to analyze interaction between G894T mutation and overweight on hypertension. Population attributable risk percent (PAR%) for them, etiologic fraction, was applied to their contribution to hypertension.

RESULTSThere was a positive interaction between G894T mutation and overweight on essential hypertension, with an index of interaction of 1.99 and attributable interaction percent of 30.76%. Their pure attributable interaction percent was 36.38%. Multiple logistic regression analysis showed that there still was positive interaction between G894T mutation and overweight on essential hypertension, adjusted for age, sex, smoking and alcohol drinking. Index of their attributable interaction was 2.85, with attributable interaction percent of 39.97%, also adjusted for the above-mentioned factors. Their pure attributable interaction percent was 46.49% and PAR% was estimated as about 15% under certain condition.

CONCLUSIONSInteraction between mutation of the 7th exon G894T of eNOS gene and overweight played an important role in essential hypertension of the studied population. Control of body weight in the population with both G894T mutation and overweight could markedly decrease their risk of hypertension.

Adult ; Exons ; Female ; Humans ; Hypertension ; etiology ; Male ; Middle Aged ; Mutation ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type III ; Obesity ; complications

OBJECTIVETo analyze the gene expression profile of degenerated cervical intervertebral disc of Sprague Dawley rats on a large scale.

METHODSDegenerated models of Sprague Dawley rats of 9 months old (degeneration group, n=9) and normal Sprague Dawley rats of 3 months old (control group, n=9) were prepared, respectively. mRNA was obtained from the cervical intervertebral disc of rats in both groups, respectively, and then labelled by Cy5 and Cy3 fluorescence respectively after reverse transcription to obtain intervertebral disc cDNA probes. cDNA probes were hybridized with BiostarR-40s gene expression profile chips and scanned by laser scanner. The results were treated with portrait analysis, standardization management, and ratio analysis with softwares.

RESULTSCompared with the rats in the control group, 9.6% (381 pieces in total) gene expression changed obviously in the rats in the degeneration group, among which, the gene expression quantities of 171 pieces increased significantly (r=the ratio of the degeneration group to the control group>2.0), 52 pieces of which had certain function. While the gene expression quantities of 211 pieces decreased significantly (r<0.5), 41 pieces of which had certain function.

CONCLUSIONSGene chip technology can be used to analyze the gene expression profile of degenerated intervertebral disc of rats in parallel, in quantity and on a large scale, which helps to testify the representative genes and protein expression, and plays an important role in clarifying the pathogenesis of degenerated intervertebral disc.

Animals ; Cervical Vertebrae ; pathology ; Disease Models, Animal ; Gene Expression Profiling ; In Situ Hybridization ; Intervertebral Disc ; pathology ; Intervertebral Disc Displacement ; genetics ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley