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Materials and Methods1. Search strategy1.1 Search strategy for the association of ACE I/D gene polymorphism with SRNS riskThe relevant studies were searched from the electronic databases of PubMed, Cochrane Library and CBM-disc (China Biological Medicine Database) on September 1, 2010. (Steroid resistant nephrotic syndrome OR SRNS) AND (Angiotensin converting enzyme OR ACE) was entered into these databases mentioned above for search. Additional articles were identified through references cited in retrieved articles.1.2 Search strategy for the relationship between ACE I/D gene polymorphism and SSNS susceptibilityA systematic literature search in PubMed, Cochrane Library and CBM-disc was carried out on September 1, 2010 using (Steroid sensitive nephrotic syndrome OR SRNS) AND (Angiotensin converting enzyme OR ACE). A search of bibliographies listed in those published studies was also conducted to identify the additional publications.1.3 Search strategy for the I/D gene distribution of ACE between SRNS and SSNSPubMed, Cochrane Library and CBM-disc were searched using (Steroid resistant nephrotic syndrome OR SRNS) AND (Steroid sensitive nephrotic syndrome OR SRNS) AND (Angiotensin converting enzyme OR ACE) as of September 1, 2010, and a search of bibliographies listed in those published studies was also performed to identify additional publications.2. Inclusion and Exclusion Criteria2.1 Inclusion and Exclusion Criteria for SRNSInclusion criteria: (1) A case–control study; (2) The outcome had to be SRNS; (3) There had to be at least two comparison groups (SRNS group vs control group); (4) Investigation was conducted in children.Exclusion criteria: (1) Review articles and editorials; (2) Case reports; (3) Investigation did not provide the detailed data of genotype distribution; (4) Preliminary results not on ACE I/D gene polymorphism or outcome; (5) Investigating the role ACE inhibitor to diseases; (6) Association of ACE I/D gene polymorphism with INS but not SRNS.2.2 Inclusion and Exclusion Criteria for SSNSInclusion criteria: (1) Case–control investigation; (2) The outcome must be SSNS; (3) There should have at least two comparison groups (SSNS group vs control group) in the study. (4) Study was performed in children.Exclusion criteria: (1) Review articles and editorials; (2) Case reports; (3) Article did not provide the detail genotype data; (4) Investigating the association of other genes with SSNS or the relation between ACE I/D gene polymorphism and other diseases. (5) Studying the role ACE inhibitor to diseases; (6) Association of ACE I/D gene polymorphism with INS but not SSNS.2.3 Inclusion and Exclusion Criteria for studies including SRNS and SSNSInclusion criteria: (1) Case–control study; (2) The outcome included SRNS and SSNS; (3) Two comparison groups (SRNS group vs SSNS) in the report was needed. (4) Investigation was implemented in children.Exclusion criteria: (1) Case reports, editorials and Review articles; (2) Investigation did not provide the detailed data of genotype distribution; (3) Results not on ACE I/D gene polymorphism or outcome; (4) Investigating the role ACE inhibitor to diseases; (5) Association of ACE I/D gene polymorphism with INS but not SRNS and SSNS.3. Data extraction and synthesisTwo investigators independently extracted the following information from each eligible study: first author's surname, year of publication and the number of cases and controls for ACE genotype. Frequency of D allele was calculated for case group and control group, from the corresponding genotype distribution. The results were compared and disagreements were resolved by discussion.4. Statistical AnalysisCochrane Review Manager Version 5 (Cochrane Library, UK) was used to calculate the available data from each investigation. The pooled statistic was counted using the fixed effects model and random effects model, respectively. Results were expressed with odds ratios (OR) for dichotomous data, and 95% confidence intervals (CI) were also calculated. P<0.05 was required for the pooled OR to be statistically significant. I2 was used to test the heterogeneity among the included studies. When a P value<0.10 indicated a significant statistical heterogeneity across studies, the results from the random effects models would be more stable when compared with those in the fixed effects model, and the final results for our study would come from those in the random effects models. In order to avoid excessive comparisons, the OR was calculated by using three methods: method 1, allele comparison (D allele vs I allele); method 2, comparing DD homozygous with the other two combinations (DD vs DI+II); method 2, comparing II genotype with the other two combinations (II vs DD+DI). A chi-square (χ2) test using a web-based program was applied to determine if genotype distributions of the control population reported conformed to Hardy-Weinberg equilibrium (HWE; P<0.05 was considered significant), and the study that the genotype distributions in the controls were significantly deviated from HWE was excluded from our sensitive analysis. All descriptive data were expressed as mean ± SD.
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