ASEAN Neurology Association (ASNA) consists of 9 member countries, Brunei, Indonesia, Lao, Malaysia, Myanmar, Philippines, Singapore, Thailand and Vietnam. Among them 2 countries are considered lower middle income, 4 as upper middle income, and 3 as high income by World Bank criteria. The life expectancy is above 75 years in Brunei and Singapore, below 60 years in Lao and Myanmar. There are a total of 1,871 neurologists in ASNA member countries which has a total of 540 million populations. This constitutes 2.2% of the world neurologists, although ASNA member countries accounts for 8.3% of the world population. Myanmar and Lao in particular, has lowest ratio of neurologist, with one neurologist serving 5 million populations in Myanmar, and 2 million populations in Lao.
Myanmar ; member ; Neurology ; Income ; Neurologist

Antibodies, Neutralizing ; Apoptosis ; Hep G2 Cells ; Humans ; Receptors, Tumor Necrosis Factor, Member 6b ; immunology ; metabolism

OBJECTIVE: To express and purify NR4A1-DNA binding domain (DBD) protein of nuclear receptors. METHODS: The fusion protein PET28a-NR4A1-DBD was constructed and purified with the nickel affinity chromatography, cation-exchange chromatography and gel filtration chromatography. RESULTS: The protein PET28a-NR4A1-DBD was mostly soluable at 24 °C. A total of 2-3 mg/L pure NR4A1 proteins were yielded in bacterial culture and the purity for final fractions of NR4A1-DBD protein were great than 95% by SDS-PAGE analysis. CONCLUSION Nickel affinity chromatography is effective to purify protein. The protein purity can be further improved by the following methods including cation-exchange chromatography and gel filtration chromatography.
Electrophoresis, Polyacrylamide Gel ; Nuclear Receptor Subfamily 4, Group A, Member 1 ; chemistry ; Recombinant Fusion Proteins ; chemistry

OBJECTIVE: To detect pathological variants of the SLC12A3 gene in a Chinese pedigree affected with Gitelman syndrome (GS). METHODS: Clinical data and peripheral blood samples of the proband and his family members were collected. All exons of the SLC12A3 gene were amplified by PCR and subjected to Sanger sequencing. RESULTS: Sanger sequencing has revealed that the proband has carried a c.486_489 delTACG (p.Ile162Met fs*8) deletion and a heterozygous c.2890C>T (p.Arg964Trp) missense variant in the SLC12A3 gene. Neither variant was reported previously and was not found among healthy controls. CONCLUSION The c.486_489delTACG (p.Ile162Met fs*8) and c.2890C>T (p.Arg964Trp) variants of the SLC12A3 gene probably underlay the GS in the proband. Above discovery has enriched the variant spectrum of GS.
China ; Gitelman Syndrome/genetics* ; Heterozygote ; Humans ; Mutation/genetics* ; Pedigree ; Solute Carrier Family 12, Member 3/genetics*

This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on.
ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Alleles ; Asian Continental Ancestry Group ; genetics ; Gene Frequency ; Genotype ; Haplotypes ; Humans

Gitelman's syndrome is an autosomal recessive disorder characterized by hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria that has recently been reported to be linked to thiazide-sensitive Na-Cl cotransporter gene mutation. In this study, we performed renal clearance studies to differentiate Gitelman's from Bartter's syndrome and to confirm the diagnosis in two patients clinically diagnosed with Gitelman's syndrome. Each patient was hydrated by 20 mL/kg body weight of oral water within 30 minutes, which was followed by intravenous half saline. When urinary flow reached 10 mL/min, samples of urine and serum were obtained to calculate the osmolar clearance, free water clearance, chloride clearance, and distal fractional chloride reabsorption. Subsequently, furosemide or hydrochlorothiazide was administered. Samples were collected and the same parameters were calculated. In our patients, chloride clearance was increased more than 10 times after furosemide administration(2.1:25.7 and 2.2:27.4 mL/min/100 mL GFR), but not increased after hydrochlorothiazide treatment(2.1:1.6 and 2.2:2.6 mL/min/100 mL GFR). And the distal fractional chloride reabsorption was significantly decreased by furosemide injection (73%:15% and 75%:4.6%), whereas hydrochlorothiazide had no effect on it(73%:63% and 75%:78%). These findings indicate that our patients have a defect in thiazide-sensitive Na-Cl cotransporter in the distal tubule, which is compatible with the pathophysiology of Gitelman's syndrome.
Alkalosis ; Bartter Syndrome ; Body Weight ; Diagnosis ; Furosemide ; Gitelman Syndrome* ; Humans ; Hydrochlorothiazide ; Solute Carrier Family 12, Member 3 ; Water

BACKGROUNDNurr1 is a member of the nuclear receptor superfamily of transcription factors. The objective of the present study was to identify novel splicing variants of the gene in neuronal and non-neuronal tissues and determine their functions.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) analysis was used to screen for Nurr1 splice variants in the adult human central nervous system (CNS) and in other tissues such as lymphocytes, and liver, muscle, and kidney cells. Functional assays of the variants were performed by measuring Nurr1 response element (NuRE) transcriptional activity in vitro.

RESULTSIn this study, the authors identified a novel splicing variant of Nurr1 within exon 5, found in multiple adult human tissues, including lymphocytes, and liver, muscle, and kidney cells, but not in the brain or spinal cord. Sequencing analysis showed the variant has a 75 bp deletion between nucleotides 1402 and 1476. A functional assay of the Nurr1-c splicing variant, performed by measuring NuRE transcriptional activity in vitro, detected a 39% lower level of luciferase (LUC) activity (P < 0.05).

CONCLUSIONA novel splicing variant of Nurr1 exists in human non-neuronal tissues and functional assays suggest that the variant may act as an alternate transcription regulator.

Adult ; Alternative Splicing ; DNA-Binding Proteins ; analysis ; genetics ; Humans ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; Transcription Factors ; analysis ; genetics

OBJECTIVETo investigate the influence of celecoxib, cycloxygenase-2 (COX-2) selective inhibitor, upon the proliferation of KB/VCR cells, and analyze the effect of celecoxib on the expression of P-glycoprotein (P-gp).

METHODSMTT method was employed to study the inhibitory effect of celecoxib on KB/VCR cells, which were divided into vincristine (VCR) group, Celecoxib group, Celecoxib + VCR group, Celecoxib + VCR + prostaglandin E2 (PGE2) group. Western blot was employed to detect the expression of P-gp, Bcl-2 and Bcl-X(L). Flow cytometry was used to evaluate the apoptosis of KB/VCR cells. All of the data were statistically analyzed using SPSS 13.0 software package.

RESULTSThe growth inhibition rate of KB/VCR cells in Celecoxib+VCR group was significantly higher than that in Celecoxib group, VCR group and Celecoxib + VCR + PGE2 group (P < 0.01). The expression of P-gp in Celecoxib + VCR group and Celecoxib group were markedly lower, compared with those in VCR group and Celecoxib + VCR + PGE2 group (P < 0.01). The expression of Bcl-2, Bcl-XL in Celecoxib+VCR group, Celecoxib+VCR+PGE2 group and Celecoxib group were significantly lower than those in VCR group (P < 0.01). The apoptosis rate of Celecoxib + VCR group, Celecoxib + VCR + PGE2 group were significantly higher than those in VCR group and Celecoxib group (P < 0.01). The apoptosis rate of Celecoxib+VCR+PGE2 group were significantly lower than those in Celecoxib+VCR group (P < 0.01).

CONCLUSIONCelecoxib could enhance the toxicity of VCR against KB/VCR cells. The mechanism probably correlates with the downregulation of celecoxib on the expression of P-gp and the increase of apoptosis.

ATP-Binding Cassette, Sub-Family B, Member 1 ; Apoptosis ; Celecoxib ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; Pyrazoles ; Sulfonamides ; Vincristine

OBJECTIVETo define the immune phenotype of colon cancer cells.

METHODSUsing a panel of 40 anti-human monoclonal antibodies (MoAbs), the cells of colon cancer HR8348 were analyzed with three-color flow cytometry after direct immunofluorescent staining.

RESULTSHR8348 cell line did not express CD2, CD3, CD4, CD5, CD7, CD8, TCR, CD10, CD11b, CD14, CD16, CD19, CD22, CD25, CD28, SmIg, CD33, CD35, CD36, CD41a, CD45, CD45RA, CD45RO, CD56, CD61, CD64, CD66b, CD69, CD71, CD117, CD122 and P-glycoprotein but expressed CD13, CD15, CD20, CD38, CD95 and HLA-DR.

CONCLUSIONThe results demonstrate that colon cancer cell line HR8348 shares some antigenic determinants with leucocyte lineage.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Antigens, CD ; analysis ; Cell Line, Tumor ; Colonic Neoplasms ; chemistry ; Humans

AIMTo assay the transcriptional activation effect of TR3 and it's deletion mutation in yeast two hybrid system.

METHODSThe total length of TR3 and TR3/delta1-690 gene was amplified by PCR method and cloned into pGBKT7 vector. Bait vector of pGBKT7-TR3 and pGBKT7-TR3/delta1-690 was transformed into AH109 competence yeast. Then self activation of the recombination vector was tested by assay the activity of beta-galactosidae.

RESULTSThe pGBKT7-TR3 and pGBKT7-TR3/AM 690 vector was successfully constructed. The filter paper containing beta-galactosidae didn't changed to blue showed that the reporter gene wasn't activationed.

CONCLUSIONTR3 and TR3/delta1-690 hadn't the activity of transcriptional activation.

Nuclear Receptor Subfamily 4, Group A, Member 1 ; genetics ; physiology ; Sequence Deletion ; Transcriptional Activation ; genetics ; Two-Hybrid System Techniques