1.Mutation analysis of the mtND4 gene associated with asthenospermia patients
Chuanlian LI ; Jiujia ZHENG ; Zong YANG ; Xuefeng HUANG ; Kexin FANG ; Zhefeng LOU ; Yonggen WU ; Longjin JIN
Chinese Journal of Urology 2011;32(1):62-66
Objective To investigate the relationship between mtND4 point mutation in sperms and asthenospermia. Methods Fifty-six asthenospermia cases and 44 control cases were collected using the WHO criterion for defining asthenospermia, the regions of mtND4 gene were amplified by using PCR of 3 pairs primers. Consequently, the point mutation, missense mutation and multiple single nucleotide polymorphisms (SNP) were analyzed by employing sequencing technology and bioinformatics tools. Results Six mutations never before identified were found. The frequency of single point mutation T10873C and T11944C in the control group were significantly higher than those in the asthenospermia group (P<0.05). Eight cases involved T10873C or T11944C among the 10 cases in control groups with missense mutations were found. But, there were only 2 cases with such mutation in the 10 asthenospermia cases with missense mutations (P<0.05). The previous 20 cases of missense mutations can be described as either multiple SNP group (with T10873C or T11944C) or nonmultiple SNP group. The percentage of a range and a plus b range of multiple SNP group of sperm was significantly higher than the non-multiple SNP group(P<0.05). Conclusions mtND4 gene mutation, especially the missense mutation may induce loss of sperm motility. The mutations of T10873C and T11944C may be useful for sperm motility or counteract the influence for the sperm motility caused by these harmful mutations.
2.Association between asthenospermia and mtDNA mutations in ND3 and ND4L genes
Chuanlian LI ; Zhefeng LOU ; Xuefeng HUANG ; Yonggen WU ; Liya ZHANG ; Jianxin Lü ; Longjin JIN
Chinese Journal of Pathophysiology 2010;26(2):362-367
AIM: To explore the molecular mechanism of asthenospermia(AST) by preliminary screening of nucleotide sequences from the ND3 and ND4L genes of mitochondrial DNA(mtDNA). METHODS: Samples from 50 AST patients and 42 age-matched normal controls were collected according to the WHO criteria. Density gradient centrifugation was applied to separate spermatozoa with different vigor. The ND3 and ND4L genes of mtDNA were amplified and sequenced directly from the extracted genomic DNA from AST patients and normal controls. The sequences were compared with revised Cambridge Reference Sequence(rCRS) to analyze the variants. RESULTS: A total of 22 nucleotide variations were found in ND3 and ND4L genes of mtDNA in asthenospermia group and control group. G10320A, A10398G and T10609C were missense mutations, while A10157G and A10313C were the reported for the first time in this study. Haplotype N in patients with AST(33/50) was higher than that in control group(14/42, P<0.05), and haplotype R9 in patients with AST(15/50) was also higher than that in control group(4/42, P<0.05) through genetic testing of ND3 gene. Rates of sperm progressive motility of haplotype F1, F2 and R9 were significantly lower than those of haplotype M and M rest. Two haplotype differences, haplotype M and N, were found in the same AST patient's spermatozoas which had different vigor. Haplotype M had stronger vigor, while haplotype N had lower vigor. By sequencing ND3 gene of mtDNA from 50 AST patients, we detected G10310A heteroplasmic mutation in 2 specimens of asthenospermia with poor and moderate motility spermatozoa, respectively. No mutation occurred in good motility spermatozoa. CONCLUSION: Haplotype of mitochondrial may have some correlation with sperm motility. The nt10398G-10400T polymorphisms may have benefit for sperm motility, whereas the mutation in nt10310A may impair sperm motility.
3.Clinical value of confocal laser endomicroscopy in diagnosis of early esophageal squamous cell carcinoma and precancerous lesions
Lijun GAO ; Tao YU ; Xiaomeng GU ; Xiuli ZUO ; Rui JI ; Changqing LI ; Chuanlian CHU ; Wenbo LI ; Yanqing LI
Chinese Journal of Digestion 2010;30(4):226-229
Objective To assess the value of confocal laser endomieroscopy (CLE) in diagnosis of early esophageal squamous cell carcinoma and precancerous lesions. Methods CLE examination was performed in 41 patients who needed further examination because of lesions in esophagus during July 2008 to April 2009. The diagnosis was made based on the features of esophageal squamous cells which was defined as low grade intraepithelial neoplasia(LGIN), high grade intraepithelial neoplasia (HGIN) and early esophageal squamous cell carcinoma (EC). Biopsy specimens were taken precisely matched to the CLE imaging sites. The result was compared with that of histopathology. Results There were 7281 eonfocal images obtained from 60 target lesions in 41 patients. The sensitivity, specificity and accuracy of CLE were 75.0%, 88.6% and 85.0% in diagnosis of LGIN, respectively, 85.7% ,92.3% and 90.0% in diagnosis of HGIN, respectively,and 88.9% ,96.1% and 95.0%, in diagnosis of EC,respectively. Conclusions It is an effective method for diagnosis of esophageal neoplastic lesions using CLE, which has high accuracy in diagnosis of HGIN and early esophageal cancer.
4. Efficacy and Safety of Prucalopride for Treatment of Chronic Functional Constipation: A Meta-analysis
Kaiyuan LI ; Yanran DU ; Yuyan ZHOU ; Chuanlian CHU ; Kaiyuan LI ; Chuanlian CHU ; Danlin SONG
Chinese Journal of Gastroenterology 2021;26(8):471-476
Background: Chronic functional constipation is a common functional gastrointestinal disorder, and the effects of traditional therapeutic drugs are suboptimal. Prucalopride is a novel 5-HT