Objective To study the characteristics of mutations of gene COL4A5 encoding type Ⅳ collagen ? 5 chain in one chinese pedigree with X-linked dominant inherited nephritis (Alport's syndrome,AS). Methods Genomic DNA was extracted from 35 members of the pedigree of Alport's syndrome. All of 51 exons of COL4A5 gene were amplified by PCR with the primers synthesized according to the published intron sequences of COL4A5. The PCR products were further analyzed by agarose gel electrophoresis and single strand conformation polymorphism (SSCP). The PCR products showing polymorphism were directly sequenced. Results PCR-SSCP analysis showed that 17 PCR products had abnormal mobility of single strand DNA. DNA sequencing analysis revealed 9 suspicious mutations. But these suspicious mutations were not be confirmed by inverse sequencing analysis. Conclusion The exon mutation of COL4A5 gene of this pedigree did not be found, and the mutations of COL4A5 gene may locate in the its introns.

Objective To detect mutation of mitochondrial DNA in a chinese pedgree with maternally inherited aminoglycoside antibiotic-induced deafness. Methods The mutation of mitochondrial DNA from all 18 family members of a chinese pedigree with maternally inherited aminoglycoside antibiotic-induced deafness was detected by PCR and DNA sequencing. Results Nine individuals in this pedigree carried A→G mutation at the 1555th bp of mitochondrial 125 rRNA, and the others did not have this mutation. Conclusion Mitochondrial DNA mutation may be one of major factors resulted in aminoglycoside antibiotic-induced deafness in this pedigree.

[Objective] To study the effects of Buyang Huanwu Decoction and Radix Astragali on astrocytes in gerbils with cerebral ischemia and reperfusion injury. [Methods] Gerbils model of cerebral ischemia was set up by occlusion of bilateral common carotid arteries. The dynamic expression of glial fibrillary acidic protein (GFAP) were determined by immunohistochemical method in reperfusion for 24 and 48 hours after 15 minutes of cerebral ischemia. [Results] Positive expression of GFAP reached a peak in reperfusion for 24 hours and was decreased by Buyang Huanwu Decoction and Radix Astragali. Positive GFAP expression was attenuated in reperfusion for 48 hours and enhanced by Buyang Huanwu Decoction and Radix Astragali increased the expression. [ Conclusion ] The regulatory effect of Buyang Huanwu Decoction and Radix Astragali on astrocytes may be one of its mechanisms in repairing nervous function after cerebral ischemia.

AIM: To study the effect of integrin ?2 on adhesion of SK-N-SH neuroblastoma cells to collagen. METHODS: Adhesion of the SK-N-SH cells to immobilized collagen was tested with various concentration of Mg~ 2+ , Ca~ 2+ and with 10 ?g/L anti-?2 monoclonal antibody (mAb) 6F1. A_ 570 was detected as adhesion cell numbers. RESULTS: Mg~ 2+ -dependent adhesion of SK-N-SH cells to type I collagen was increased significantly, with peak adhesion at concentration of 1 mmol/L Mg~ 2+ . A_ 570 with or without Mg~ 2+ was 0.59?0.03 and 0.25?0.01 respectively (P

Objective To investigate effects of angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism on endothelial function in elderly patients with essential hypertension. Methods Seventy-three elderly patients with uncomplicated essential hypertension and 55 normal elderly people as control were recruited into the study. Their humeral arterial endothelial vasodilatation function was studied with high-resolution ultrasonography and ACE I/D genotypes were determined by polymerase chain reaction (PCR) technique. Results Endothelium-dependent vasodilatation function of the three-genotype subgroups of the elderly patients with essential hypertension was significantly lower than that in the controls with the same genotypes (5.5±1.9 vs 11.9±1.3 in Ⅱ genotypa, 4.7±2.0 vs 10.9±1.6 in ID genotype and 2.9±1.9 vs 9.4±2.6 in DD genotype, with all P<0.01 ). In both groups of hypertensives and normotensives, humeral arterial endothelium-dependent vasedilatation function was significantly lower in those with DD genotype than that in those with Ⅱ genotype (2.9±1.9 vs 5.5±1.9 in hypertensive group and 9.4±2.6 vs 11.9±1.3 in control group, both P<0.05 ). Multivariate linear regression analysis showed that endothelium-dependent vasodilatation function in hypertensive elderly people correlated in linearity with their ACE genotype (P<0.01 ). Conclusions ACE gene I/D polymorphism is associated with abnormal humeral arterial endothelium-dependent vasodilatation function of humeral artery in patients with essential hypertension.

In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.

OBJECTIVETo detect mutation in the rhodopsin gene (RHO) in a Chinese family with autosomal dominant retinitis pigmentosa (ADRP).

METHODSA total of 25 family members from a Chinese family were investigated. All the subjects were examined clinically by direct funduscopy, perimetry and vision test. Evaluation of the proband included electroretinography (ERG). Genomic DNA was extracted using standard method. The complete coding regions of RHO were amplified by polymerase chain reaction (PCR) and the PCR products were subjected to automatic DNA sequencing.

RESULTS512 C>T (P171L), a recurrent missense mutation was detected in the proband. All 12 affected subjects in the family were heterozygous for the mutation. The affected individuals had night blindness at the age of 5-6 years. They had relatively severe impairment of visual acuity and suffered a gradual loss of peripheral visual field at the age of 20-30 years. And they went blind at the age of 40-50 years. Rod and cone ERG were not detectable in the proband.

CONCLUSIONA recurrent missense mutation, 512C>T (P171L), was detected in a Chinese family with ADRP.

Adolescent ; Adult ; Base Sequence ; China ; DNA Mutational Analysis ; Family Health ; Female ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Retinitis Pigmentosa ; genetics ; pathology ; Rhodopsin ; genetics

OBJECTIVE: To explore the characteristics of copy number variation (CNV) within the Y chromosome azoospermia factor (AZF) region in patients with spermatogenesis disorders in the Shenzhen area. METHODS: A total of 123 patients with spermatogenesis disorders who had visited Shenzhen People's Hospital from January 2016 to October 2022 (including 73 patients with azoospermia and 50 patients with oligozoospermia) and 100 normal semen males were selected as the study subjects. The AZF region was detected with multiplex ligation-dependent probe amplification (MLPA), and the correlation between the CNV in the AZF region and spermatogenesis disorders was analyzed using the chi-square test or Fisher's exact test. RESULTS: 19 CNV were detected among 53 patients from the 223 samples, including 20 cases (27.40%, 20/73) from the azoospermia group, 19 cases (38%, 19/50) from the oligozoospermia group, and 14 cases (14%, 14/100) from the normal control group. In the azoospermia, oligozoospermia, and normal control groups, the detection rates for CNV related to the AZFa region (including AZFab and AZFabc) were 5.48% (4/73), 2.00% (1/50), and 0 (0/100), respectively. The detection rates for the AZFb region (including the AZFbc region) were 6.85% (5/73), 0 (0/50), and 0 (0/100), respectively. The detection rates for gr/gr deletions in the AZFc region were 2.74% (2/73), 6.00% (3/50), and 9.00% (9/100), respectively, and those for b2/b4 deletions in the AZFc region were 2.74% (2/73), 10.00% (5/50), and 0 (0/100), respectively. The detection rates for complex rearrangements in the AZFc region were 6.85% (5/73), 18.00% (9/50), and 3.00% (3/100), respectively. Statistical analysis showed no significant difference in the detection rate of gr/gr deletions between the three groups (Fisher's Exact Test value = 2.712, P = 0.249); the differences in the detection rate of b2/b4 deletions between the three groups were statistically significant (Fisher's Exact Test value = 9.489, P = 0.002); the differences in the detection rate of complex rearrangements in the AZFc region between the three groups were statistically significant (Fisher's Exact Test value = 9.493, P = 0.006). In this study, a rare AZFa region ARSLP1 gene deletion (involving SY86 deletion) was detected in a patient with oligozoospermia. CONCLUSION CNV in the AZFa and AZFb regions have a severe impact on spermatogenesis, but partial deletion in the AZFa region (ARSLP1 gene deletion) has a minor impact on spermatogenesis. The b2/b4 deletion and complex rearrangement in the AZFc region may be risk factors for male infertility. The gr/gr deletion may not serve as a risk factor for male infertility in the Shenzhen area.
Humans ; Male ; Azoospermia/genetics* ; DNA Copy Number Variations ; Oligospermia/genetics* ; Infertility, Male/genetics* ; Y Chromosome