| Objective:To research into the optimum experimental conditions of the Leu262Val Polymorphism of PARL gene detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Methods:Optimize PCR program by touchdown PCR (TD-PCR); Set series of gradient to research the main factors influencing the experimental conditions of Parl gene, such as:annealing temperature, Primer concentration, Enzyme system, Enzyme digestion time and so on. Set different temperature gradients to analysis ordinary PCR and TD-PCR, set four final concentration gradients for the primer(0.2umol/L、0.4umol/L、0.6umol/L、0.8umol/L) to conduct amplification. In enzyme system, the quantity of incision enzyme does not change, and the quantity of PCR product is respectively set as5μl、7.5μl、10μl、12.5μl、15μl. Enzyme digestion time is12h or4h. Observe the result through gel electrophoresis. Results:(1) During the process of TD-PCR preceding ordinary PCR annealing temperature; Nonspecific product quantity decreases, product quantity is more and is easily identified. Amplified fragments length is545bp (2)When primer concentration is0.4μmmol/L, it is clear in the definition of Amplified Stripes, and the dimer of primers is little.(3)Add10μl amplified product to enzyme system, clear enzyme stripes can be obtained.(4) When the temperature is37℃, after four hour digestion, the result of electrophoresis is clear. Conclusion: TD-PCR precedes ordinaty PCR in amplification efficiency, the optimum primer concentration is0.4μmmol/L, The experimental cycle number is32cycles; Enzyme system is the20μl system10μl product by1μl(10U) incision enzyme, When the time of enzyme digestion is4h, the effect of enzyme digestion is the best. Objective:To explore the association of the Leu262Val polymorphism of PARL gene and pre-diabetes, diabetes mellitus paitents among Lahu people in Lincang District,Yunnan Province,China. Methods:The Leu262Val polymorphism of PARL gene was detected by PCR-RFLP in108people with pre-diabetes (PD),52people with Diabetes mellitus(DM) and308normal controls (NC). The genotypes and allele frequency distributions were compared among different groups of people, at the same time, the related clinical and biochemical indicators were also compared. According to the survey by questionnaire, Logistic regression analysis was used to estimate the risk markers of pre-diabetes and diabetes mellitus. Results:(1) There were no obvious differences in genotypes and allele frequency distributions among PD, DM and NC (P>0.05) in Lahu people. There were no obvious statistical differences in genotypes and allele frequency distributions between male and female at this point.(2)The levels of the proportion of women,age, SBP、DBP、ALT、AST、BUN、CREA、 UA、TC、HDL、LDL-C、FPG in PD were obviosly higher than NC (P<0.05); The levels of the proportion of women,age, ALT、AST、BUN、CREA、UA、TC、HDL-C、 LDL-C、FPG in DM were obviously higher than NC(P<0.05), The CREA、FPG in DM were higher than PD (P<.05)(3) There were no obvious differences in the clinical indicators among the patients carrying the PARL gene Leu262Val polymorphism genetypes in NC、PD and DM (P>0.05).(4) Logistic regression analysis showed Age, BMI, high blood pressure, food cooking oil, the degree of education, and family annual income may be relevant factors for PD and DM. Conclusion:There may be no obvious association between the Leu262Val polymorphism of PARL gene and PD、 DM in Lahu People of Lincang District, Yunnan Province, China. ALT、AST、BUN、 CREA、TC、LDL-C may be related with PD and DM. Age, BMI, high blood pressure, food cooking oil, the degree of education, and family annual income may be relevant factors for PD and DM. Objective:To discuss the association of the Leu262Val polymorphism of PARL gene and type2diabetes among Han people in Kunming District,Yunnan Province, China. Methods:The Leu262Val polymorphism of PARL gene was detected by PCR-RFLP in145people with diabetes,and111normal controls (NC). The genotypes and allele frequency distributions were compared, at the same time, the related clinical and biochemical indicators were assessed. Results:(1) There were no obvious statistical differences in genotypes and allele frequency distributions between DM and NC (P>0.05) in the nation. There were no clear statistical differences in genotypes and allele frequency distributions between male and female at this point (P>0.05).(2)The levels of age, BMI、SBP、DBP、TG、LDL-C. CREA in NC were obviously lower than DM; but the level of HDL-C was higher than DM (P<0.05)。(3) Respectively compare the clinical and biochemical indicators of carrying the polymorph of different genotypes, There were no clear differences in clinical indicators between GG and CC+GC (P>0.05). Conclusion:the Leu262Val polymorphism of parl gene may be not related with DM in Han people in Kunming District; the SBP、DBP、TG、LDL-C may be related with DM. Objective:To explore the discrepancy of the Leu262Val polymorphism of PARL gene between the Han and Lahu people of Yunnan province. Methods:According our previous research result, select360Lahu people(308normal controls,52diabete patients),256Han people(111normal controls,145diabetes patients).Compare and analyse the genotype and allele frequency of the Leu262Val polymorphism of PARL gene. Assess related clinical and biochemical indicators. Results:1) There were differences in genotype and allele frequency of the samples between Lahu and Han People, There is statistical significance in the difference (P<.001).(2) There were obvious differences in genotype frequencies and allele frequencies between Lahu and Han People in the normal glucose controls.There is statistical significance in the difference (P<0.001).(3) There were differences in genotype and allele frequencies between Lahu and Han People in diabetes patients, and there is statistical significance (P<0.05).(4) In the normal controls of the two nations, the levels of age, ALT、 BUN、CREA、TC、HDL-C、LDL-C in Han people were higher than Lahu people (P<0.05);(5) In diabetes patients of the two nations, the levels of ALT and TG in Han people were higher than Lahu people; the levels of SBP, AST、CREA、UA were lower than Lahu people (P<0.05). Conclusion:There may exist racial diversity in genotype and allele frequencies with the Leu262Val polymorphism of PARL gene between the Han and Lahu people. |
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