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OBJECTIVE@#To explore the clinical feature and gene variant for two cases of primary male infertility caused by severe asthenospermia and to analyze the etiology of the disease.@*METHODS@#Genomic DNA of peripheral blood samples of patients and their parents was extracted and gene variant analysis of the patients was conducted by using whole exome sequencing. Suspected pathogenic variant was verified by Sanger sequencing and pathogenic analysis.@*RESULTS@#Whole exome sequencing showed that the DNAH1 gene of patient 1 had two heterozygous variants of c.2016T>G(p.Y672X) and c.6017T>G (p.V2006G). The DNAH1 gene of patient 2 had a homozygous variant of c.2610G>A(p.W870X), which were inherited from his father and mother, respectively. According to American College of Medical Genetics and Genomics standards and guidelines, the c.2016T>G (p.Y672X) and c.2610G>A (p.W870X) varaints of DNAH1 gene were predicted to be pathogenic (PVS1+PM2+PM3+PP3).@*CONCLUSION@#The two patients of multiple morphological abnormalities of the sperm flagella may be caused by DNAH1 gene variant, which has resulted in primary male infertility.
Subject(s)
Humans , Male , Dyneins/genetics , Genomics , Infertility, Male/genetics , Mutation , Sperm Tail/pathology , Exome SequencingABSTRACT
Objective To explore the significance of detection serum Retinol-Binding Protein and AnnexinA2 in diabetic ne-phropathy.Methods According to 24-UAER level,80 diabetic patients with suspected DN were devided into three groups, that was A (25 cases),B (26 cases)and C (29 cases)group,and control group (NC)was 25 cases.HbA1c,BUN,Scr,UA and RBP leves were detected by the standard method.The expression level of Annexin A2 were detected by real-time PCR. Results There were obvious differences of HbA1c,BUN,Scr,UA and RBP levels among three groups and NC group (t=4.64~13.65,P0.05),there were differences of RBP (t=15.26,P<0.05).To HbA1c,BUN,Scr UA and RBP,there were differences be-tween A groups and C groups (t=5.26~25.33,P<0.05),there were the same results between B groups and C groups (t=4.02~18.33,P<0.05).To expression level of AnnexinA2 there were differences among three group and NC group ((t=13.45~24.25,P<0.05)).There exist positive correlation between the expression of AnnexinA2 and RBP level (r=0.95, P<0.01).Conclusion Combined detection of serum RBP and expression level of AnnexinA2 may be a sensitive and early phase indicator for impairment of renal function for diagnosis of diabetic nephropathy.
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BACKGROUND:Chinese herbs for kidney nourishment can promote the proliferation and differentiation of spermatogonial stem cel s. OBJECTIVE:To verify the effect of seminiferous capsule extract on spermatogonial stem cel proliferation. METHODS:Spermatogonial stem cel s were isolated from the testis of male mice and synchronized by serum-free medium fol owed by an addition of 10, 50, 100 mg/L seminiferous capsule extracts. After 24 hours of culture, viability, proliferation and cel cycle of spermatogonial stem cel s were observed. RESULTS AND CONCLUSION:Compared with the control group, seminiferous capsule extracts promoted the cel number, viability and proportion at S stage. The number of BrdU-labeled spermatogonial stem cel s was increased significantly after intervention with seminiferous capsule extracts, especial y at the concentration of 50 mg/L. These findings indicate that seminiferous capsule extracts can promote the proliferation and viability of spermatogonial stem cel s.
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BACKGROUND:To establish a rapid and effective method to obtain sufficient spermatogonial stem cels that can meet the clinical need is urgent to be solved in the spermatogonial stem cel transplantation. OBJECTIVE:To study the effect of rhodiola polysaccharide on the proliferation of spermatogonial stem celsin vitro. METHODS:Under sterile conditions, spermatogonial stem cels and Sertoli cels were isolated from the testis of mice, and spermatogonial stem cels were seeded onto the feed layer of Sertoli cels. Then, the co-cultured cels were assigned into experimental group 1 (simple cel culture medium), experimental group 2 (cel culture medium containing 150 mg/L rhodiola polysaccharide) and experimental group 3 (cel culture medium containing 150 mg/L rhodiola polysaccharide, 1 U/L leukemia inhibitory factor and 10 μg/L glial cel line-derived neurotrophic factor). After 7 days of co-culture, flow cytometry was used to detect cel proliferation in vitro, and cel viability and positive expression of GFRa-1, Thy-1 and C-kit were calculated. RESULTS AND CONCLUSION:After 7 days of co-culture, the cels grew rapidly and presented with colony and thyrsiform growth, and the number of cel masses increased significantly, al of which were in line with the proliferative features of spermatogonial stem cels. The GFRa-1, Thy-1 and C-kit proteins were expressed in the cel membrane and cytoplasm, mainly in the cel membrane. The viability of spermatogonial stem cels and positive expression of GFRa-1 and Thy-1 were ranked as folows: experimental group 3 > experimental group 2 > experimental group 1, and there were significant differences between groups (P < 0.05). The positive expression of C-kit had no difference between experimental groups 1 and 2, but it was significantly higher in the experimental group 3 than the other two groups (P < 0.05). These findings indicate that rhodiola polysaccharide used alone or combined with leukemia inhibitory factor and glial cel line-derived neurotrophic factor can enhance the proliferative ability of spermatogonial stem celsin vitro.