Objective To investigate the clinical features and distribution of G types of rotavirus diarrhea in infants and children in Suzhou and Maanshan areas.Methods 1267 stool specimens were collected from children under 5 years with acute diarrhea. Enzyme-linked immunosorbant assay (ELISA) and nested polymeras chain reaction (Nested-PCR) were used to detect rotavirus and serotypes.Results In 1267 stool specimens, 378 were detected RV positive, and serotype G was the prevalent type in the two cities with number of 250 and the G3 and G1 types were the most prevalent which accounted for 40%(100/250) and 44.40 % (111/250), respectively. While the G2 ,G4, G9 types and mixed infections were 8,3,3,4 and the un-identified type was 8.40%(21/250). G1 type was the most prevalent in Maanshan are which accounted for 58.54% in all cases,while the G3 type accounted for the most of case(47.85%) in Suzhou area.Conclusions The results indicated that rotavirus was the most important etiologic agent in Maanshan and Suzhou areas,G1 and G3 were the most prevalent types,respectively. J Appl Clin Pediatr,2005,20(3):208-210

Objective To explore the incidence and associated factors of aortic artery calcification (AAC) by lateral lumbar X-ray score in maintenance hemodialysis (MHD) patients.Mehtods A total of 155 MHD patients with complete clinical data in our hospital were enrolled in the study.Lateral lumbar X-ray score of the abdominal aorta was used to determine AAC in MHD patients.Results Aortic calcification was most severe in front of the fourth lumbar segment and ameliorated in higher lumbar levels.63.63％ of MHD patients presented visible calcification in the abdominal aorta,and 28.39％ had severe calcification with more than three segments.Age (OR=1.094,P＜0.01),dialysis vintage (OR=1.013,P=0.022),triglyceride (OR=1.261,P=0.030) and phosphate level (OR=1.324,P=0.023) were risk factors of abdominal aorta calcification,however serum albumin level (OR =0.239,P=0.013) was protect factor of aortic calcification.Conclusions Incidence of AAC is quite high in MHD patients and associated with increasing of age,duration of hemodialysis,serum triglyceride,phosphate level and plasma albumin.The semi-quantitative X-ray method of determining vascular calcification is less expensive and may be widely available clinically.

The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.
Amino Acid Sequence ; Base Sequence ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Exons ; Introns ; Orchidaceae ; genetics ; Phylogeny ; Plant Leaves ; genetics ; Plant Proteins ; genetics ; Plants, Medicinal ; genetics ; Sequence Alignment ; Sequence Homology, Amino Acid

Adult ; Heroin Dependence ; drug therapy ; prevention & control ; Humans ; Male ; Methadone ; therapeutic use ; Patient Compliance ; Treatment Outcome

Random amplified microsatellite polymorphism (RAMP) markers were used to access the genetic diversity among 112 samples of nine populations of Dendrobium officinale Kimura et Migo. Using 16 informative primers, 123 bands were amplified and 86 (69.92%) were polymorphic. The polymorphic bands from three to eight could be detected for each RAMP primer, with a mean of 5, indicating abundant genetic diversity among populations. Genetic similarity coefficients ranged from 0.250 to 0.813. UPGMA dendrogram illustrated 9 populations clustered into 3 groups, and the cluster pattern showed correlation with the locations of the D. officinale populations. These results were supported by the previous conclusions that were achieved by other molecular markers, and RAMP is proved to be effective for evaluating the genetic diversity of wild populations of Dendrobium officinale.
Cluster Analysis ; DNA Primers ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; methods

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.
Base Sequence ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Orchidaceae ; classification ; Plants, Medicinal ; classification

BACKGROUNDGenetic factors are believed to play a role in the individual susceptibility to chronic obstructive pulmonary disease (COPD). A single nucleotide polymorphism (SNP) of tumour necrosis factor-alpha (TNF-alpha) has been reported but inconsistent results may arise from different populations and phenotypes of COPD. There are only a few published studies of interleukin-13 (IL-13) SNPs on COPD. The SNPs of TNF-alpha and IL-13 have not been studied in the Chinese population. This research was conducted to study the frequencies of IL-13 gene promoter 1055 (IL-13-1055) and TNF-alpha gene-308 polymorphisms in the patients with COPD and to investigate the effect of those genetic polymorphisms on COPD in the Chinese population.

METHODSA cohort of COPD patients and age matched controls were recruited from an inpatient hospital service in Beijing. Venous blood was obtained and genomic DNA was extracted from peripheral blood monocytes using standard method. Genomic DNA was used as a template for amplification by polymerase chain reaction (PCR) to determine the polymorphism at -1055 in the IL-13 gene promoter region. PCR restriction fragment length polymorphism (RFLP) was used to determine polymorphisms in the TNF-alpha gene-308 position. The products were investigated by sequence analysis also.

RESULTSOne hundred and eleven COPD patients and 97 controls were studied. Seventy-five cases were current smokers in COPD patients and 36 were current smokers in controls. The frequencies of TT genotype in the IL-13 gene promoter region were 11.7% (13/111) in the COPD group and 13.4% (13/97) in the controls (P = 0.713). However, the OR value of TT genotype was significantly increased to 6.4 (95% CI 1.62 - 25.39) in the smokers with COPD. TT genotype was also positively related to family history of COPD, OR = 7.7 (95% CI 1.37 - 43.80). The frequencies of A allele in the TNF-alpha gene were 5.9% in COPD and 3.1% in controls (P = 0.131). The OR value of A allele was 5.0 (95% CI 1.011 to 25.059) in smokers with COPD.

CONCLUSIONSThere is no significant difference in the frequencies of the TT genotype of IL-13-1055 or the A allele of the TNF-alpha between Han Chinese patients with COPD versus control. Thus, it does not appear that these SNPs are independent factors in COPD for Han nationality in Beijing. However, these SNPs may increase the risk of COPD among smokers.

Adult ; Aged ; China ; ethnology ; Female ; Genotype ; Humans ; Interleukin-13 ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Pulmonary Disease, Chronic Obstructive ; genetics ; immunology ; Tumor Necrosis Factor-alpha ; genetics

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

Simple sequence repeat (SSR) was used to investigate the genetic diversity and structure of Dendrobium officinale. A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads. Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism. These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1. The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively. The polymorphic information content (PIC) of each SSR locus varied from 0.437 to 0.829 with an average of 0.702. Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D. officinale related species. In addition, 500 tissue culture plantlets of D. officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs. The results showed that SSR technique is a feasible, simple and inexpensive method for determining adulterants in germplasm identification.
DNA Primers ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Genetic Variation ; Microsatellite Repeats ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Species Specificity

OBJECTIVETo analyze the molecular epidemiological characteristics of HIV-1 subtype CRF01_AE strains being prevailed among HIV/AIDS in Zhejiang province in 2009.

METHODSA total of 303 subjects were identified by stratified random sampling among HIV infected individuals in Zhejiang province in 2009. Gag fragments of the HIV-1 strains were amplified by nested polymerase chain reaction from the DNA extracted from whole blood of HIV-1 infected individuals. PCR products were sequenced and analyzed by phylogenetic method.

RESULTSA total of 132 HIV-1 subtype CRF01_AE sequences were identified from the 225 samples that sequenced successfully, accounting for 58.67% (132/225). A total of 90.91% (120/132) CRF01_AE strains infected HIV/AIDS were transmitted mainly by sexual contacts. A total of 65.91% (87/132) of the cases infected by heterosexual route and 25.00% (33/132) by homosexual route. There were three main clusters in the phylogenetic tree. Pairwise DNA distance within three groups was 0.037 ± 0.011, 0.034 ± 0.008 and 0.047 ± 0.010, which has statistical significance (P < 0.05). Distribution of the sequence of homosexual behavior infected individuals was relatively concentrated in clusters one (96.97%, 32/33), and crossed with heterosexual behavior infectors, and presented the close relations with strains from Jiangsu province, Zhengzhou of Henan province, Liaoning province, Shijiazhuang of Hebei province.

CONCLUSIONThe CRF01_AE strains were the dominant subtypes among HIV infected individuals. The majority of the CRF01_AE infected cases had high risk sexual behavior. The heterosexual infected cases were more than homosexual cases. The circulating status of CRF01_AE strains in homosexual population was relatively independent, but also had evidence of transmission from man who have sex with man to heterosexual population.

Acquired Immunodeficiency Syndrome ; epidemiology ; virology ; Adolescent ; Adult ; China ; epidemiology ; Female ; Genetic Variation ; HIV-1 ; classification ; genetics ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Phylogeny ; Young Adult