1.Exploration of the dynamic responses of Sertoli cells to depletion of spermatogonial stem cells in mouse testis
Na AN ; Lifang YUAN ; Shanlin WANG ; Zongyue LI ; Feng WANG ; Baochang ZHU
Acta Laboratorium Animalis Scientia Sinica 2014;(5):7-12
Objective To explore the dynamic responses of Sertoli cells to depletion of spermatogonial stem cells by busulfan.Methods After intraperitoneal injection of 15, 30 or 44 mg/kg busulfan to mice, the spermatogenesis and the expression of GDNF, PLZF, Nanog and GFRɑ1 mRNA were assessed by real-time quantitative PCR at 5 and 28 days after the busulfan treatment.Results Glial cell line-derived neutrophic factor ( GDNF ) was significantly increased and showed a dose-dependent trend at 5 days after busulfan treatment, but no significant difference was seen in the expression of promyelocytic leukemia zinc finger(PLZF) and GDNF family receptorα-1(GFRα1).The testicular histology also appeared no significant difference at 5 days after busulfan treatment.At 28 days after busulfan treatment, the relative expression lev-els of GDNF, PLZF, Nanog and GFRɑ1 mRNA were drastically increased.Morphological observation showed that spermat-ogenesis damages became even more severe as the busulfan dose increased.Conclusions Sertoli cell response to the de-pletion of spermatogonia occurs as early as the fifth day after busulfan treatment.Production of GDNF in Sertoli cells shows a compensatory increase, which may stimulate spermatogonial stem cells to accelerate their self-renewal, reflected by the enhancing expression of Nanog and PLZF, and ultimately promote the restoration of spermatogenesis.
2.Identification and functional analysis of combined oxidative phosphorylation deficiency 28 gene mutation
Ping SHI ; Yiping CHENG ; Zongyue LI ; Shuping WANG ; Yingzhou SHI ; Yiming JI ; Li FANG ; Jiajun ZHAO ; Ling GAO ; Chao XU
Chinese Journal of Internal Medicine 2022;61(12):1324-1329
Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.
3.Distribution of vascular endothelial growth factor gene polymorphism in Uyghur Ethnic patients with urolithiasis in south Xinjiang
Yuefu HAN ; Qinzhang WANG ; Guofu DING ; Biao QIAN ; Jiangping WANG ; Yinglong LI ; Zhao NI ; Xinmin WANG ; Shunming XIE ; Wenxiao WANG ; Zongyue CHEN ; Guodong ZHU ; Shiqi JI ; Yujie WANG ; Niwaer AN ; Guanglu SONG ; Hui WEI ; Qunying ZHANG
Chinese Journal of Urology 2010;31(9):601-603
Objective To determine vascular endothelial growth factor(VEGF)-460 gene polymorphism in Uyghurs and its relationship to urolithiasis in south Xinjiang. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),gene sequencing and genetic analysis methods were used in 200 urolithiasis patients of Uyghurs, and 200 healthy Uyghurs. Results The distribution of genotype and allele had no significant difference between urolithiasis patients and normal controls (P>0. 05). The frequencies for the CC,TT and CT genotypes in patients with urolithiasis and normal controls were 1.5 %, 29.0 %, 69.5 % and 0. 5 %, 27.5 %, 72.0 %, respectively. The frequencies for C and T allele were 36.2%,63.7% and 36.9% ,63.1%, respectively. Conclusions The results of VEGF-460 gene polymorphisms indicate no significant relationship between patients with turolithiasis and normal controls in Uyghurs in south Xinjiang,which may not be urolithiasis susceptibility genetic locus.
4.Research process in molecular genetics of Gitelman syndrome
Zongyue LI ; Chao XU ; Ling GAO
Chinese Journal of Endocrinology and Metabolism 2020;36(4):348-351
Gitelman syndrome(GS) is an autosomal recessive genetic disease caused by mutations in the SLC12A3 gene located in chromosome 16q13. The incidence of GS is 1-10∶40 000. SLC12A3 encodes thiazide-sensitive sodium-chloride cotransporters(NCC) which play key roles in Na + and Cl - reabsorption. GS is characterized by hypokalemic metabolic alkalosis in combination with significant hypomagnesaemia and low urinary calcium excretion. There are some correlations between genotypes and phenotypes. In previous studies, more than 500 mutations have been identified and some of them have been functionally analyzed. We review genetic mutations and functional studies related to GS as well as the relationship between genotypes and phenotypes, and summarize the research process in molecular genetics of GS.
5.Current Status, Opportunities and Challenges of Rare Endocrine and Metabolic Diseases
Huixiao WU ; Zongyue LI ; Chao XU ; Jiajun ZHAO
JOURNAL OF RARE DISEASES 2023;2(3):319-324
The rare endocrine and metabolic diseases, due to their varieties, face many challenges in the study of clinical diagnosis, pathogenesis, and treatment. In the past a couple of years, the research on rare endocrine and metabolic diseases has been gradually improved. The diagnosis has made great progress. The research into molecular mechanism of rare endocrine and metabolic diseases has significantly advanced. The effort in exploring the breakthroughs and progress in therapeutic methods based on the pathogenesis of the diseases has also made. This article provides a brief overview of the current status of research into diagnosis, mechanism, and treatment of rare endocrine and metabolic diseases. In addition, the article points out the problems and challenges and proposes future possibilities.