| Objective To investigate whether the human Genomic DNA, extracted by PUREGENE DNA Isolation Kit, Phenol-chloroform protocol or Miller salting out method, could all be used to detect the DNA polymorphism retrospectively, and compare the quantity, concentration, efficiency as well as cost of DNA extraction by each method.Methods human Genomic DNA was extracted from 300μl Peripheral blood samples by PUREGENE DNA Isolation Kit(n=42), Phenol-chloroform protocol(n= 14) and Miller salting out method(n= 11), respectively. The quantity and concentration of extracted DNA was determined using a spectrophotometer and efficiency as well as cost of each method was observed. The extracted DNA was fractioned on a 1.5 % Agarose gel per-stained with ethidium bromide and visualized under UV rays and was used as a template for PCR.Results The extracted Genomic DNA, whatever demonstrated by PUREGENE DNA Isolation Kit, Phenol-chloroform protocol or Miller salting out method, could all be used to detect the DNA polymorphism, the quantity of extracted DNA obtained by PUREGENE DNA Isolation Kit(5.1±0.9μg) and Phenol-chloroform protocol (6.8±1.6μg) were more than those got by Miller salting out method(2.3μg, Median), (P < 0.01); However, there was no statistical difference in concentration of extracted DNA among each method (P >0.05). the PUREGENE DNA Isolation Kit was more effective(P < 0.01) and expensive(P < 0.01) compared to other two methods.Conclusion Each method could all be used to extract human Genomic DNA for DNA polymorphism investigation. The PUREGENE DNA Isolation Kit was easier, more effective and expensive than others; and the quantity of extracted DNA obtained by Phenol-chloroform protocol were more than those got by Miller salting out method . An appropriate method should be chosen on budget and requirement of experimentation.Objective To investigate whether a C/A single nucleotide polymorphism at position -160 from the transcription start site of E-cadherin gene promoter is associated with transitional cell carcinoma of urinary bladder (TCCB).Methods A hospital-based case-control study was performed on 50 patients with TCCB (male 38 and female 12, age form 33 to 72, mean 53.8±12.3) and 50 normal controls (male 31 and female 19, age form 22 to 69, mean 48.9±13.1). Genomic DNA was extracted from blood samples of the subjects. Genotypes were determined using PCR based restriction fragment length polymorphism (PCR-RFLP) technique.Results The A allele frequencies at -160 position of E-cadherin gene promoter were significantly higher in TCCB than in normal controls (P<0.01) ,and A-allele carriers had a higher relative risk of TCCB (OR=4.16, 95% CI1.74-9.93) compared to C-only carriers. AC and AA genotypes had an increased risk of TCCB (OR =3.48, 95% CI 1.26-9.63 and OR=4.91, 95% CI 1.79-13.45, respectively) compared to CC genotypes. The A allele frequencies at the same position were also significantly higher in invasive TCCB than in superficial carcinoma (P <0.05) . However, there was no statistical difference in the A allele frequency between high pathological grade and low pathological grade of TCCB (P >0.05).Conclusion The -160 C/A single nucleotide polymorphism of E-cadherin gene promoter is associated with TCCB. This single nucleotide polymorphism may serve as a prognostic marker of TCCB.Objective To investigate the association between an Insertion/Deletion polymoiphism in +234 position of Intron 1 of CDH1 gene and TCCB risk retrospectively.Methods A hospital-based case-control study was performed on 130 patients with TCCB and 60 normal controls. Genomic DNA was extracted from blood samples of the subjects. Genotypes were determined using PCR and DNA sequencing technique. Linkage analysis of the -160 C/A SNP and the +234 Insertion/Deletion polymorphism in CDH1 gene was performed by PHASE software.Results The association between -160 C/A SNP and TCCB was the same as desribed in section 2. a 5'-CCGTGCCCCAGCC-3' Insertion/Deletion polymorphism in +234 position of Intron 1 of CDH1 gene was detected by DNA sequencing technique. Its genotype was homozygous for 5'-CCGTGCCCCAGCC-3' allele insertion(I/I), homozygous for 5'-CCGTGCCCCAGCC-3' allele deletion(D/D) and heterozygous for 5'-CCGTGCCCCAGCC-3' allele insertion/repeat insertion(I/2I), respectively. The repeat insertion frequencies at +234 position of Intron 1 of E-cadherin gene were significantly higher in TCCB than in normal controls (P<0.05 ) . However, there was no statistical difference in the repeat insertion frequency between neither superficial and invasive TCCB nor high pathological grade and low pathological grade of TCCB (P >0.05). I/2I carriers had a higher relative risk of TCCB (OR=2.22, 95% CI 1.17-4.21) compared to I/I carriers.Haplotype scanning performed by PHASE software shown that the frequency of -160A/A-+234I/2I genotype were significantly higher in TCCB cases than in normal controls (x~2= 6.50,P<0.05) .while the -160C/C-+234I/I frequency were significantly lower than normal controls (x~2=7.09,P<0.05 ) , and -160A/A-+234I/2I carriers had ahigher relative risk of TCCB (OR=6.61, 95%CI 1.93-22.64) compared to -160C/C-+234I/I carriers.Conclusion The -160 A SNP and +234 5'-CCGTGCCCCAGCC-3' repeat insertionof E-cadherin gene promoter is associated with TCCB. Those may serve as a prognostic marker of TCCB.Objective Toevaluate the technical feasibility and clinical efficacy of retroperitoneoscopic subcapsular nephrectomy for infective non-functioning kidney with dense perinephric adhesion.Materials and Methods: Twelve patients were subjected to retroperitoneoscopic subcapsular nephrectomy, of whom 8 were males and 4 females, aged from 35-66 years old (mean 41.4±9.2 min), 7 for left side and 5 for right side, 5 with tuberculous pyonephrosis, 7 with severe pyonephrosis, including 4 patients who received percutaneous nephrostomy, 2 ureteral stent drainage previously on the diseased kindey for 2-12 weeks. The operating time, blood loss, postoperative intestinal function recovery, complications during operation and the operative efficacy were observed. Dissociation and ligation of renal pedicle is the most important step, in which renal capsule is cut near to renal hilum using Harmonic Scalpel and the fat tissue around the renal hilum is dissected carefully, and then the renal pedicle was ligated and divided using an Endoscopic Linear Cutter when the renal pedicle tissues have been separated to adequate thickness.Results: All operations were performed successfully with none of the cases converted to open surgery. The average operating time was 45-120 min, (mean 82.9±22.3 min), average blood loss was 30-75 ml, (mean 51.4±12.2 ml) and postoperative intestinal function recovery time was 12-48 h. No complications occurred during and after operation. A follow-up of 1-15 months revealed satisfactory short-term results. Conclusion: Retroperitoneoscopic subcapsular nephrectomy can be safely applied to the removal of infective and heavily adhesive non-functioning kidney with the advantages of minimal trauma and blood loss, quick recovery compared to open subcapsular nephrectomy.
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