Study on clinical manifestation, genotype and genetic characteristics of two Kennedy disease pedigrees / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical manifestations, genotypes, and genetic characteristics of two pedigrees with Kennedy disease.</p><p><b>METHODS</b>The clinical data of the patients from two Kennedy disease families were collected. The numbers of trinucleotide CAG repeats in exon 1 of the androgen receptor gene were determined by DNA sequencing and repeat fragment analysis.</p><p><b>RESULTS</b>Family A was composed of 58 individuals in 4 generations. The proband had onset at 39 years old. There were two Kennedy disease patients in family B which included 61 individuals in 5 generations. The two patients had onset at 39 and 41 years old, respectively. All the three patients displayed limbs and bulbar muscular weakness because of the damage of lower motor neurons. They had androgen insensitivity syndrome in common, and showed mild or moderate increase in serum creatine kinase level. The electromyogram showed wild damage in anterior horn of spinal cord. Muscle biopsy displayed neurogenic muscular atrophy. The numbers of the CAG repeat expansion in the androgen receptor gene of the three patients were 49, 48, and 47, respectively. X-linked recessive mode of inheritance was demonstrated by pedigree analysis in the two families.</p><p><b>CONCLUSION</b>Kennedy disease usually occurs in mid-adulthood man. The clinical features are the weakness and wasting of limbs and bulbar muscles. Genetic analysis contributes to diagnosis and identification of carriers, and is beneficial to genetic counseling and prenatal diagnosis.</p>

Analysis of β-thalassemia mutations in Guangdong province / 中国地方病学杂志

Objective To investigate the prevalence and spectrum of β-thalassemia mutations in C, uangdong province, and provide a reference for prenatal diagnosis and genetic counseling in this population. Methods Three thousand two hundred and forty-seven blood samples were randomly selected from Guangzhou and 2984 blood samples from Shenzhen from January in 2005 to January in 2009. PCR and reverse dot blot hybridization (RDB) were adopted for detection of β-thalassemia mutations in Guangzhou and Shenzhen city. Results Seven hundred and fifty-one individuals in Guangzhou were found to have β-hemoglobin gene mutations, the detection rate was 23.13%(751/3247); 10 different mutations were identified, namely CD41-42(-TCTT), IVS-Ⅱ-654(C→T), -28(A→G), CDI7(A→T), CD71-72(+A), 13E, IVS-I-1(G→T), CD43(G→T), -29(A→G), CDI4-15(+G), which accounted for 42.53% (336/790) ,25.19% (199/790), 12.66% (100/790), 10.89% (86/790) ,3.29% (26/790), 2.15%(17/790), 1.27%( 10/790), 1.14%(9/790) ,0.51%(4/790) ,0.38%(3/790), respectively; the most common mutation was CD41-42(-TCTT), which accounted for 42.53%(336/790). In Shenzhen, 179 individuals were found to have β-thalassemia mutations, the detection rate was 6.00% (179/2984); 8 different mutations were identified excluding CD43 (G→T) and CD14-15 (+G); the most common mutation, however, was IVS-lI--654(C→T), which accounted for 40.44% (74/183). Conclusions The β-thalassemia mutations in Guangdong province are not only frequent, but also obviously heterogeneous, and the mutations differ from region to region. CD41-42 (-TCTT),ⅣS-Ⅱ-654(C→T), -28(A→G), CD17(A→T) were the 4 predominant mutations.

B16-F10 melanoma cells and cell culture supernatant enhance angiogenesis in mouse ischemic limb / 生理学报

Generation of therapeutic angiogenesis to enhance vascularization in the ischemic tissues is a method for treating ischemic tissues in atherosclerotic cardiovascular artery disease. The chemokine stromal cell-derived factor-1 (SDF-1) and its receptor (CXC chemokine receptor 4, CXCR4) play a critical role in the process of post-natal neovascularization. The SDF-1-CXCR4 axis is a potential mechanism for the treatment of ischemic limb. Here, we investigated the role of CXCR4 in bone marrow cells (BMCs) in neovascularization induced by tumor cells and the supernatant of culture media in a murine hind-limb ischemia model which was made by resecting femoral artery and vein. After the injection of mouse melanoma cells B16-F10 (1x10(6) cells in 0.1 mL at the operation day, s.c.) into the abdomen or the cell culture supernatant (0.1 mL/d for 21 d after operation, i.m.) into the ischemic abductor muscle, the CXCR4 positive BMCs were analyzed by flow cytometry. The perfusion of the ischemic limb was evaluated by laser Doppler perfusion imaging (LDPI) on 7, 14 and 21 d after vascular injury operation. Capillary endothelial alkaline phosphatase (AP) was stained to quantify the presence of capillaries, and histological method was used to evaluate the capillary density as a measure of neovascularization in ischemic tissues. The proportions of CXCR4 positive BMCs were notably higher in ischemic limb injected with tumor cells or the supernatant compared to those in the control group (P<0.05). Injection of tumor cells or the supernatant resulted in significantly improved perfusion as measured by LDPI perfusion ratios on 7, 14 and 21 d after femoral artery and vein resection in mice, compared to the controls (P<0.05). Tissue samples harvested from the lower calf muscle at day 21 demonstrated increased capillary densities in mice receiving tumor cells (0.81+/-0.13) or the supernatant (0.63+/-0.05), compared with those in control group (0.44+/-0.09, P<0.05). In conclusion, the injection of B16-F10 tumor cells or the supernatant induces the increase of CXCR4 positive cells in BMCs and the improvement of in vivo neovasculogenesis in mouse ischemic limb.

Polymorphism of DXS15, CA13, CA22 loci in Guangdong normal population / 中国实验血液学杂志

The aim of this study was to investigate the polymorphism of microsatellite repeats DXS15, CA13, CA22 tightly linked to FVIII gene in Guangdong population and its practical value in genetic diagnosis for hemophilia A. The polymerase chain reaction (PCR) and capillary electrophoresis (CE) methods were adopted to test the variability of the 3 microsatellite repeat in Guangdong females, including 111 females, 222 X chromosomes for detecting DXS15 polymorphism; 87 females, 174X chromosomes for detecting CA13 polymorphism; 94 females, 188 X chromosomes for detecting CA22 polymorphism. The results indicated that 11 alleles corresponding to DXS15 were found at this locus with size ranging from 140 to 160 bp. The polymorphism information content (PIC) of this microsatellite repeat was 0.82, heterozygosity was 82%. Six alleles corresponding to CA13 were found, with a size from 145 to 155 bp, and PIC was 0.56, heterozygosity was 56.2%. Four alleles corresponding to CA22 were found with size ranging from 79 to 85 bp, and PIC was 0.41, heterozygosity was 50%. It is concluded that in contrast to the information about Caucasian, the polymorphism of these 3 microsatellites differs from race to race, and region to region. DXS15, CA13 and CA22 are highly polymorphic genetic markers useful for linkage analysis of haemophilia A, which may play a vital role in detection and prenatal diagnosis for hemophilia A.

Expression of Class I integrase gene in Acinetobacter baumannii and drug-resistance / 中南大学学报(医学版)

OBJECTIVE@#To explore the expression level of class I integrase (intI 1) mRNA in Acinetobacter baumannii from biofilm cells and planktonic cultured cells ,and to analyze the drug-resistance of Class I integron positive strains.@*METHODS@#Acinetobacter baumannii were collected from hospitals,and Class I integron strains were screened by gene amplification. Total RNA of Class I integron positive strains was extracted, and the intI1 mRNA expression in the bioflim cells and planktonic cultured cells was measured by RT-PCR. Susceptibilities to antibiotics of Class I integron positive strains were also examined.@*RESULTS@#The intI1 gene mRNA was expressed under 2 conditions, and the mRNA expressed in the biofilm cells was about 4 times higher than that in the planktonic cultured cells. Among the 64 strains of Acinetobacter baumannii, 46 strains were Class I integron positive strains. The antibiotic resistance of intI1 gene cassette-positive strains was higher than that of gene cassette-negative strains.@*CONCLUSION@#The intI1 gene mRNA can be up-regulated in Acinetobacter baumannii biofilm cells.Class I integron plays an important role in drug resistance. It is much easier to capture gene cassettes for bacteria under biofilm condition.

Polymorphisms of human papillomavirus type 81 L1 gene in patients with cervical lesions in Guangdong / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the positivity rates and genetic polymorphism of human papillomavirus (HPV) type 81 (HPV81) L1 gene in patients with cervical cancer in Guangdong.</p><p><b>METHODS</b>HPV L1 genes fragment in 1200 positive cervical samples detected with HC-II for high-risk HPV DNA were amplified by HPV-specific PCR with consensus primers, and the L1 genes of HPV81 were sequenced for mutation analysis.</p><p><b>RESULTS</b>Four HPV81-positive cases were found in the total of 1200 cases for the positivity rate of 0.33%. Point mutations of HPV81 L1 gene were found in all of the samples.</p><p><b>CONCLUSION</b>Compared with the HPV81 prototypes published in GenBank, several new point mutations are found in the L1 genes of the HPV81 in Guangdong, and these mutations are close to those found in Japan.</p>

Research on the procedure for recovery and species identification of Legionella from surface environmental water / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish a set of procedure for recovery and species identification of Legionella from the surface environmental water.</p><p><b>METHODS</b>Forty-four water samples were collected in eight parks of Guangzhou city from August to November in 2006. The bacteriologic examination was performed by cultivation on BCYEalpha plate, and 108 presumptive Legionella colonies were picked and their homogeneous relationship was analyzed by using an amplified fragment length polymorphism (AFLP) method. Species identification was carried out by latex agglutination test, biochemical characterization, analysis of cellular fatty acids composition, 16 S rRNA gene and mip gene sequencing.</p><p><b>RESULTS</b>Legionella was recovered among 27 (61.36%) samples of all eight parks, and 31 different strains were identified from those 108 presumptive Legionella isolates by AFLP method, including 20 strains of L. pneumophila, five strains of L. feeleii, four strains of L. longbeachae, one strain of L. oakridgensis and one strain of L. sainthelensi, and L. pneumophila could be easily differentiated by phenotypic and biochemical characteristics, latex agglutination test or analysis of the cellular fatty acids composition . However, uncertain factors were existing in those phenotypic identification methods as compared to the sequence analysis.</p><p><b>CONCLUSION</b>The taxonomic analysis of the Legionellae family should be dependent on the 16 S rRNA gene or mip gene.</p>

Beta-thalassemia major caused by compound heterozygosity for +40 to +43(-AAAC), IVS-2-654 (C to T) and codon 41/42 (-TCTT) / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report the analysis of a rare beta-thalassemia ternary heterozygote [+40 to +43(-AAAC)*CD41/42(-TTCT)*IVS-2-654] causing beta-thalassemia major in a Chinese.</p><p><b>METHODS</b>Using PCR-ASO probe hybridization analysis to scan 17 known types of beta-thalassemia mutations, and gene cloning and DNA sequencing to identify the underlying causative mutation.</p><p><b>RESULTS</b>Reverse dot blot (RDB) analysis showed that the patient's beta-globin gene had three mutations: +40 to +43(-AAAC), CD41/42(-TCTT) and IVS-2-654(C to T). Beta-globin gene cloning and sequencing proved that, the two deletions of +40 to +43(-AAAC) and CD41/42(-TCTT) co-existed on the same chromosome, and the other homologous chromosome had an IVS-2-654 (C to T) mutation. So the patient is a compound heterozygote of [+40 to +43(-AAAC)*CD41/42 (-TCTT)]/IVS-2-654 (C to T) leading to beta-thalassemia major.</p><p><b>CONCLUSION</b>The triple mutation of [+40 to +43(-AAAC)*CD41/42(-TCTT)/N] is a new genotype of beta-thalassemia in Chinese.</p>

Surgical treatment for lumbar discogenic low back pain / 中南大学学报(医学版)

OBJECTIVE@#To explore the curative effect of surgical treatment for lumbar discogenic low back pain.@*METHODS@#Forty-two patients with lumbar discogenic low back pain were treated with disc excision and lumbar interbody fusion: 12 of them were treated with posterior lumbar interbody fusion (PLIF), 30 with afront lumbar interbody fusion (ALIF). The clinical results were evaluated by pre- and post-operative visual analogue scale (VAS), and the fusion rates of the lumbosacral spine were evaluated by X-ray.@*RESULTS@#The average follow-up period was 18 months, ranging from 6 to 36 months. Low back pain basically disappeared in 11 patients treated with PLIF, with mild back pain in one patient; VAS after the operation decreased significantly in comparison with that preoperatively (P<0.01). Low back pain basically disappeared in 28 patients treated with ALIF; VAS after the operation decreased significantly in comparison with that preoperatively (P<0.01). The fusion rate was 90% in patients treated with PLIF, and 93% in patients with ALIF.@*CONCLUSION@#Disc excision and interbody fusion is effective in the treatment of lumbar discogenic low back pain, but the operation indications should be paid attention to.