Cloning and prokaryotic expression of LEN-5 β-lactamase gene / 中国人兽共患病学报

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.

GJB2 gene mutation in deaf patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect the GJB2 gene mutation in patients with autosomal-recessive deafness, and analyze the relationship between clinical phenotype and gene mutation.</p><p><b>METHODS</b>Forty-two patients were examined clinically by pure tone audiometry, acoustic impedance and auditory brainstem response. The complete coding region of the GJB2 gene was amplified by polymerase chain reaction (PCR) and the PCR products were subjected to automatic DNA sequencing.</p><p><b>RESULTS</b>Two cases had homozygous mutation of 235delC. One of them had sensorineural hearing loss while the other had mixed hearing loss. Heterozygous mutation of 176del16bp was detected in a pair of twins who had mixed hearing loss. The 109G to A, 79G to A and 341A to G mutations were observed in both the patients and the controls.</p><p><b>CONCLUSION</b>Homozygous 235delC mutation is one of the pathogeni c mutations which could occur in patients with mixed hearing loss. The heterozygous 176del16bp mutation combined with environmental factor may cause hearing loss. The 109G to A, 79G to A and 341A to G variants were considered to be polymorphisms of the GJB2 gene.</p>

Effects of angiotensin converting enzyme gene polymorphism on endothelial function in patients with essential hypertension / 中华全科医师杂志

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.

The relationship between the effect of amlodipine on carotid intimal-medial thickness and angiotensin converting enzyme genotypes in elderly hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the association between the anti-atherosclerotic effects of amlodipine and angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism in elderly essential hypertensive (EH) patients.</p><p><b>METHODS</b>A total of 220 EH patients were treated with amlodipine (2.5 - 10 mg, once daily) for twelve months and complete data were obtained from 208 patients with genotypes of II (n = 90), ID (n = 91) and DD (n = 27). The indices of carotid arterial were compared before and post amlodipine treatment in patients with identical genotype and among different ACE genotypes and each genotype post therapy.</p><p><b>RESULTS</b>The carotid mean intimal-medial thickness (MIMT) was slightly decreased in EH patients with ID and DD genotypes and significantly decreased in EH patients with II genotype (0.96 +/- 0.12 vs. 0.92 +/- 0.13, P < 0.01) compared to pre-treatment values. The decreased degree of MIMT (DeltaMIMT) in II genotype was significantly higher in II genotype than those in ID or DD genotype (0.05 +/- 0.03 vs. 0.01 +/- 0.02, 0.01 +/- 0.03 respectively, P < 0.01). The post treatment plaque score (PS) in patients with II genotype was significantly reduced (4.85 +/- 2.51 vs. 3.90 +/- 2.36, P < 0.05). Multivariate linear regression analysis showed the baseline SBP, the decreased degree of SBP (DeltaSBP) and the II genotype were the major factors affecting the DeltaMIMT.</p><p><b>CONCLUSION</b>Hypertensive patients carrying II genotype ACE genotype are the best responders for the anti-atherosclerotic effects of amlodipine.</p>

A recurrent rhodopsin gene missense mutation in a Chinese family with autosomal dominant retinitis pigmentosa / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect mutation in the rhodopsin gene (RHO) in a Chinese family with autosomal dominant retinitis pigmentosa (ADRP).</p><p><b>METHODS</b>A 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.</p><p><b>RESULTS</b>512 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.</p><p><b>CONCLUSION</b>A recurrent missense mutation, 512C>T (P171L), was detected in a Chinese family with ADRP.</p>

Effects of bu yang huan wu decoction on astrocytes after cerebral ischemia and reperfusion / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion.</p><p><b>METHOD</b>Cerebral ischemia model in gerbils was produced by ligating bilateral common carotid artery. The dynamic expressin of GFAP were determined by immunochemistry after cerebyal ischemia for 15 min followed by reperfusion for 24 hours and 48 hours.</p><p><b>RESULT</b>GFAP positive reactions reached a peak after cerebral ischemia for 15 min followed by reperfusion for 24 hours. Bu Yang Huan Wu Decoction decreased the expression. GFAP positive reactions decreased after cerebral ischemia for 15 min followed by reperfusion for 48 hours, Bu Yang Huan Wu Decoction increased the expression.</p><p><b>CONCLUSION</b>The regulation of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion may be related to repairing process after cerebral ischemia.</p>

A Study on Angiotensin Converting Enzyme Gene Polymorphisms in Essential Hypertension Patients with Left Ventricular Hypertrophy / 中国医师杂志

Objective To explore the relationship between the polymorphism of angiotensin converting enzyme (ACE) gene and essential hypertension complicated with left ventricular hypertrophy (LVH). Methods ACE gene I/D polymorphism in 150 healthy subjects, 80 essential hypertensive(ET)patients with LVH and 152 ET patients without LVH was detected by PCR. Left ventricular mass (LVM) was measured by echocardiography, and then left ventricular mass index (LVMI) was calculated. Results The frequencies of the ACE D allele in the ET patients with or without LVH were 0.493 and 0.514, respectively, and significantly higher than those in the healthy subjects (P

Mutation Analysis for Mitochondrial DNA in a Chinese Pedigree with Maternally Inherited Aminoglycoside Antibiotic-Induced Deafness / 中国医师杂志

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.

Effects of Buyang Huanwu Decoction and Radix Astragali On Astrocytes in Gerbils with Cerebral Ischemia and Reperfusion Injury / 广州中医药大学学报

[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.