Effect of Smo SiRNA-mediated Hedgehog Signaling Pathway Inhibition on Palatal Fusion / 生物医学与环境科学（英文）

We used Smo siRNA to inhibit hedgehog signaling pathway in embryonic day (E) 13 palatal shelves in organ culture. SiRNA 4 was chosen as the most efficient from four synthesized Smo siRNAs. Palatal shelf fusion rate of 4 μg/mL cyclopamine group was the lowest and significantly lower than that of blank control group (P<0.05), and that of siRNA 4 group was also lower than that of blank control group (P=0.183). At 48 h after transfection, Smo protein level of siRNA 4 group was 64.8% lower than that of blank control group (P<0.05), and Gli1 protein level of 4 μg/mL cyclopamine group was 68.9% lower than that of blank control group (P<0.05). Hedgehog signaling pathway inhibition decreased palatal fusion in organ culture, probably owing to downregulation of Smo and Gli1 proteins.

Association between polymorphism of IRF6 rs2235371 locus and nonsyndromic cleft lip with or without cleft palate / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between polymorphism of interferon regulatory factor 6 (IRF6) gene rs2235371 locus and nonsyndromic cleft lip with or without cleft palate in Chinese population.</p><p><b>METHODS</b>Blood samples from 106 patients and their parents and 129 controls and their parents were collected. The polymorphism of IRF6 rs2235371 locus was determined with PCR-restriction fragment length polymorphism (PCR-RFLP) method. Case-control analysis, transmission disequilibrium test(TDT), haplotype-based haplotype relative risk analysis (HHRR) and family-based association test (FBAT) were carried out.</p><p><b>RESULTS</b>By case-control analysis, no significant difference was found in the frequencies of GG, GA and AA genotypes of rs2235371 locus between the patient group and control group (P> 0.05), but there was a significant difference in allelic frequencies (P< 0.05). There was also a significant difference in genotype and gene frequencies of rs2235371 variant between family members from cleft lip only group and control group. However, in cleft lip with cleft palate group, no such difference was observed. TDT analysis suggested a linkage in the presence of disequilibrium (chi-square=5.56, P=0.024). Results of HHRR analysis (chi-square=5.115, P=0.024) and FBAT (Z=2.218, P=0.027) also indicated an association between IRF6 rs2235371 variant and the risk of NSCL with or without cleft palate.</p><p><b>CONCLUSION</b>Genetic polymorphism of IRF6 gene rs2235371 locus is associated with NSCL with or without cleft palate.</p>

Genome-wide and interaction linkage scan for nonsyndromic cleft lip with or without cleft palate in two multiplex families in Shenyang, China / 生物医学与环境科学（英文）

<p><b>OBJECTIVES</b>To identify the loci involved in nonsyndromic cleft lip with or without cleft palate (NSCL/P) in Northern Chinese people in Shenyang by using genomewide and interaction linkage scan.</p><p><b>METHODS</b>Two multiplex families in Shenyang from North China were ascertained through probands with NSCL/P. Blood of every member was drawn for DNA extraction and analysis. Genotypes were available for 382 autosomal short tandem repeat (STR) markers from the ABI Prism Linkage Mapping Set version 2.5. Linkage between markers and NSCL/P was assessed by 2-point parametric LOD scores, multipoint-heterogeneity parametric LOD scores (HLODs), and multipoint nonparametric linkage score (NPL).</p><p><b>RESULTS</b>The initial scan suggested linkage on Chromosomes 1, 2, and 15. In subsequent fine mapping, 1q32-q42 showed a maximum multipoint LOD score of 1.9(empirical P=0.013) and an NPL score of 2.35 (empirical P=0.053). For 2p24-p25, the multipoint NPL increased to 2.94 (empirical P=0.007). 2-locus interaction analysis obtained a maximum NPL score of 3.73 (P=0.00078) and a maximum LOD score of 3 for Chromosome 1 (at 221 cM) and Chromosome 2 (at 29 cM).</p><p><b>CONCLUSION</b>Both parametric and nonparametric linkage scores greatly increased over the initial linkage scores on 1q32-q42, suggesting a susceptibility locus in this region. Nonparametric linkage gave a strong evidence for a candidate region on chromosome 2p24-p25. The superiority of 2-locus linkage scores compared to single-locus scores gave additional evidence for linkage on 1q32-q42 and 2p24-p25, and suggested that certain genes in the two regions may contribute to NCSL/P risks with interaction.</p>

No association of the A2756G polymorphism of methionine synthase gene with nonsyndromic cleft lip with or without cleft palate / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the association of the A2756G polymorphism of the methionine synthase (MS) gene with nonsyndromic cleft lip with or without cleft palate (NSCL/P) in Chinese.</p><p><b>METHODS</b>Ninety-seven NSCL/P case-parent triads were selected as the case group. One hundred and four healthy subjects and their biological parents were selected as control group. For all subjects the A2756G polymorphism of the MS gene was examined by PCR-RFLP method.</p><p><b>RESULTS</b>There was no statistical difference in genotype and allele frequencies for MS A2756G variants among family members between case group and control group. The GG genotype was not detected in the offsprings and mothers. The odds ratio and confidence interval of genotype AG in offspring, father and mother were 1.78(0.74-4.34), 0.80(0.36-1.79) and 1.26(0.54-2.93) respectively. The odds ratio and confidence interval of allele G in offspring, father and mother were 1.70(0.78-3.73), 0.88(0.49-1.75), and 1.23(0.59-2.60) respectively. The G allele did not increase the risk of NSCL/P. Transmission disequilibrium test (TDT) analysis yielded no evidence of linkage disequilibrium (chi-square=0.034,P>0.05). The results of haplotype-based haplotype relative risk (HHRR) analysis (chi-square=0.03,P>0.05) and family-based association tests (FBAT) (Z=0.186, P>0.05) failed to show association between the MS A2756G variant and the risk of NSCL/P.</p><p><b>CONCLUSION</b>The A2756G polymorphism of the MS gene was not associated with NSCL/P in Chinese in the present study.</p>

Plasma homocysteine and gene polymorphisms associated with the risk of hyperlipidemia in northern Chinese subjects / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To examine the relationship between occurrence of hyperlipidemia, plasma homocysteine and polymorphisms of methylenetetra hydrofolate reductase (MTHFR) gene and methionine synthase (MS) gene.</p><p><b>METHODS</b>A total of 192 hyperlipidemia patients were selected and divided into hypercholesterolemia group, hypertriglyceridemia group, and combined hyperlipidemia group. Another 208 normal individuals were selected as control. Total plasma homocysteine (tHcy) concentration was measured by high-performance liquid chromatography (HPLC). Lipid profiles were measured for all subjects. The polymorphisms of MTHFR gene C677T and MS gene A2756G were analyzed by PCR-RFLP.</p><p><b>RESULTS</b>The tHcy concentration in the combined hyperlipidemia patients was significantly higher than that in the control (15.95 micromol/L vs 13.43 micromol/L, P < 0.05). The prevalence of hyperhomocysteinemia (HHcy) in the combined hyperlipidemia group was significantly higher than that in the control (42.2% vs. 23.0%, P = 0.015), with the odds ratio (OR) of 3.339 (95% CI: 1.260-8.849). The hyperlipidemia patients with HHcy had a higher concentration of total cholesterol (TC) than that in the normal tHcy patients (5.67 +/- 0.95 mmol/L vs. 5.47 +/- 0.92 mmol/L, P=0.034). There was no significant difference in genotype or allele frequencies of MTHFR C677T between the hyperlipidemic and control groups. The hyperlipidemia patients with MTHFR CT/TT genotype had a higher concentration of triglyceride (TG) than those with CC genotype (2.24 +/- 1.75 mmol/L vs 1.87 +/- 0.95 mmol/L, P < 0.05). Individuals with CT/TT genotype had a higher concentration of tHcy than those with 677CC genotype both in the hyperlipidemia group (12.61 +/- 1.24 micromol/L vs. 11.20 +/- 1.37 micromol/L, P < 0.05) and in the control group (14.04 +/- 1.48 micromol/L vs. 12.61 +/- 1.24 micromol/L, P < 0.05). The percentage of MS 2756 GG/AG genotype in the combined hyperlipidemia group was significantly higher than that in the control (26.7% vs. 13.0%, P = 0.012), with the OR of 3.121 (95% CI: 1.288-7.651). The hyperlipidemia patients with MS 2756AG/GG genotype had a higher concentration of TC (5.87 +/- 0.89 mmol/L vs. 5.46 +/- 0.93 mmol/L, P < 0.05) and LDL-C (3.29 +/- 0.81 mmol/L vs. 2.94 +/- 0.85 mmol/L, P < 0.05) than those with AA genotype. However, individuals with 2756AG/GG genotype showed no significant difference in tHcy among those with AA genotype.</p><p><b>CONCLUSION</b>HHcy and MS A2756G mutation may be the risk factors for combined hyperlipidemia. Further study is needed to confirm the role of HHcy and MS A2756G mutation in the development of hyperlipidemia.</p>

Relationship between polymorphism of cystathionine beta synthase gene and congenital heart disease in Chinese nuclear families / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the relationship between polymorphism of cystathionine beta synthase (CBS) gene and development of congenital heart disease (CHD).</p><p><b>METHODS</b>One hundred and twenty-seven CHD case-parent triads were recruited from Liaoning Province as patient group, and 129 healthy subjects without family history of birth defect were simultaneously recruited as control group together with their biological parents. For all subjects the polymorphism of CBS gene G919A locus was examined by PCR-ARMS method.</p><p><b>RESULTS</b>The frequencies of three genotypes (w/w, w/m, and m/m) in control group were 27.2%, 58.4%, and 14.4%, respectively, with no significant difference in gender. A significant difference in the allele frequency was found between CHD patients and controls, the wild allele frequency was 67.9% in patients and 55.7% in controls. CHD parents' genotype distribution was significantly different from that in controls. Further comparison of each type of CHD showed that genotype frequencies in several CHD subtypes were significantly different from those in their corresponding controls. The results of TDT analysis showed that no allele transmission disequilibrium existed in CHD nuclear families.</p><p><b>CONCLUSIONS</b>CBS gene G919A mutation is associated with the development of CHD, and the mutated allele may decrease the risk of CHD.</p>

Relationship between polymorphism of methylenetetrahydrofolate dehydrogenase and congenital heart defect / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the relationship between G1958A gene polymorphism of methylenetetrahydrofolate dehydrogenase (MTHFD) and occurrence of congenital heart disease (CHD) in North China.</p><p><b>METHODS</b>One hundred and ninety-two CHD patients and their parents were included in this study as case group in Liaoning Province by birth defect registration cards, and 124 healthy subjects (age and gender matched) and their parents were simultaneously selected from the same geographic area as control. Their gene polymorphism of MTHFD G1958A locus was examined with PCR-RFLP, and serum folic acid and homocysteine (Hcy) levels were tested with radio-immunoassay and fluorescence polarization immunoassay (FPIA).</p><p><b>RESULTS</b>There existed gene polymorphism at MTHFD G1958A locus in healthy subjects living in North China. The percentages of GG, GA, and AA genotype were 57.98%, 35.57%, and 6.45% respectively, and the A allele frequency was 24.23%, which was significantly different from Western population. No difference was observed when comparing genotype distribution and allele frequency between the case and control groups, so was the result from the comparison between genders. The A allele frequency of arterial septal defect patients' mothers (10.87%) was significantly lower than that of controls (28.15%) (P=0.014), with OR=0.31 (95% CI: 0.09-0.84), and no difference in the other subgroups. The percentage of at least one parent carrying A allele in arterial septal defect subgroup (43.48%) was significantly lower than that in controls (69.64%) (P=0.017), with OR=0.34 (95% CI: 0.12-0.92). The analysis of genetic transmission indicated that there was no transmission disequillibrium in CHD nuclear families. Their serum folic acid level was significantly higher than that of controls (P=0.000), and Hcy level of the former was higher than that of the latter with no statistical significance (P>0.05). Serum Hcy and folic acid levels of mothers with gene mutation were lower than those of mothers with no mutation.</p><p><b>CONCLUSION</b>No significant difference of genotype distribution and allele frequency existed between CHD patients and healthy population. MTHFD G1958A mutation in parents (particularly in mother) can decrease the risk of arterial septal defect in offspring. The possible mechanism of protection might be mutation, which can increase MTHFD enzyme activity, folic acid metabolism and homocysteine remethylation, and decrease Hcy level.</p>

Polymorphism of methionine synthase gene in nuclear families of congenital heart disease / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype.</p><p><b>METHODS</b>One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method.</p><p><b>RESULTS</b>In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+) in case fathers (5.0%) was apparently lower than that in the control (9.1%, P = 0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-) with OR 0.26 (95% CI: 0.11-0.60).</p><p><b>CONCLUSION</b>MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.</p>

Effects of dietary intervention on hyperlipidemia in eight communities of Beijing, China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To examine the effect of community-based dietary intervention on hyperlipidemia.</p><p><b>METHODS</b>A total of 180 hyperlipidemia individuals with TG>2.26 mmol x L(-1) (200 mg x dL(-1)) and/or TC>5.72 mmol x L(-1) (220 mg x dL(-1)) were selected from 428 eligible subjects in eight communities of Beijing. They were randomly divided into intervention group (n=108) and control group (n=72). Dietary intervention was provided for the intervention group for 6 months. Information on dietary intakes, physical examinations and blood samples was collected. Serum lipids were assayed at baseline and endpoint of the study period.</p><p><b>RESULTS</b>Respective decrease in dietary intake of total calories, fat, cholesterol and cooking oil by 13.62%, 24.75%, 24.40%, and 22.43%, in the intervention group was observed. The percentages of total calories from fat, carbohydrate and protein appeared to be desirable after study. Reduced body weight and BMI were also observed. There was a respective 5.61% and 7.06% decrease in total serum cholesterol and low-density lipoprotein cholesterol in the intervention group, while no significant changes were found in the control group.</p><p><b>CONCLUSIONS</b>Community-based dietary intervention can effectively improve dietary patterns, control body weight, and decrease the levels of total serum cholesterol and low-density lipoprotein cholesterol.</p>

The effect of dietary intervention on lipidemia in school-aged children / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of dietary intervention on lipidemia in school-aged children.</p><p><b>METHODS</b>The levels of serum lipids profile were detected in 316 school children aged 7 - 11, from those the subjects of dietary intervention were selected by total cholesterol level above 4.26 mmol/L or low density lipoprotein cholesterol level above 2.23 mmol/L. The subjects were randomly divided into intervention group (120) and control group (40). Children in intervention group were fed with low-cholesterol and low-saturated fatty acid diet, and the control group with normal diet. The duration of intervention was three months. Before and after the intervention, the food intakes, health-related questionnaire and physical examination (height, weight, skinfolds thickness and so on) in the two groups were studied.</p><p><b>RESULTS</b>Compared with the control group, serum cholesterol levels of children under intervention were not significantly changed (TC: 4.64 vs 4.68 mmol/L, P > 0.05; LDL-C: 2.66 vs 2.62 mmol/L, P > 0.05), but the apoA(1) level increased from 1,378.4 mg/L to 1,441.3 mg/L (P < 0.05). There were no changes for any serum lipids indexes in the control group while the dietary intakes of energy, cholesterol and SFA decreased markedly in the intervention group, with the percent of energy from fat decreased from 40.7% to 31.2% and SFA below to 10% (7.7%). Along with the increase of the scores of knowledge on health among children under intervention, the living and eating habit improved (the total scores increased from 24.6 to 27.4, P < 0.05). The increase of height was not significantly different between the two groups.</p><p><b>CONCLUSION</b>With the family-based high-risk intervention strategy on the dietary adjustment, the knowledge on health, living and eating habit could be effectively improved in children with lipidemia. However, further research about the intervention effect on the serum cholesterol levels by strengthening the interventional degree, needs to be further studied.</p>