Trichinella spiralis nudix hydrolase (TsNd) gene encoding a 46 kDa protein was expressed in Escherichia coli and the potential of recombinant TsNd protein (rTsNd) as an antigen for the serodiagnosis of trichinellosis was investigated by ELISA and compared with those of ELISA with T. spiralis muscle larval excretory–secretory (ES) antigens. The sensitivity of both ELISA was 100% (30/30), for the detection of anti-Trichinella IgG antibodies in sera of the experimentally infected mice, and the specificity of rTsNd-ELISA and ES-ELISA was 100% (54/54) and 98% (53/54), respectively (P>0.05). Serum anti-Trichinella antibodies were firstly detected by rTsNd-ELISA at 14 days post infection (dpi), then continued to increase with a detection rate of 100% at 36 dpi. The anti-Trichinella antibody levels at different times after infection were statistically different (P<0.05). The results showed that the rTsNd might be a potential candidate antigen for specific serodiagnosis of trichinellosis. But, it needs to be further evaluated with sera of the patients with trichinellosis and other helminthiasis.

Proteomics plays important roles in Chinese medicine research at post-genomics era. Its research idea and methods are beneficial for elucidating some elemental features of Chinese medicine. At present, Chinese medicine proteomic studies are mainly focusing on the syndromatology and medical herbal pharmacology. However, there are still some problems, the most important matter was that most of the results were merely the superficial delineations. Further research should put emphasize on the unremitting and penetrating study of proteomics, molecular biology and bioinformatics integrally for illuminating Chinese medicine theory deeply to promote the modernization of Chinese medicine research.
Computational Biology ; Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; methods ; Proteomics ; Research

OBJECTIVETo determine the chroma value of sintered IL1-IL5 zirconia materials in comparison with the Vita In-Ceram YZ color shade.

METHODSFive types of shading dental zirconia ceramics with color gradient were prepared by adding Fe2O3, CeO2, and Bi2O3 to the zirconia powder, and their chroma values were determined using a spectrophotometer and the color difference was calculated.

RESULTSThe chroma value ranges were L: 67.76-77.78, a: -2.19-3.80, and b: 12.13-25.01. Slight deltaE was found between IL1 and LL1, IL2 and LL2, and IL3 and LL3. The deltaE between IL4 and LL4 could be compensated by veneering porcelain, whereas deltaL between IL5 and LL5 could not be compensated in this manner.

CONCLUSIONShading dental zirconia ceramics can be prepared by addition of metal oxides with color similar to the Vita In-Ceram YZ color shades to match that of the veneering porcelain in clinical practice.

Color ; Dental Porcelain ; Dental Veneers ; Metal Ceramic Alloys ; Prosthesis Coloring ; methods ; Spectrophotometry ; Zirconium

OBJECTIVETo obtain the functional information of AY358935 gene.

METHODSThe properties, subcellular location, and structure of AY358935 protein, and the expression profile of AY358935 gene were analyzed by bioinformatics software and the biological functions of the gene were predicted. AY358935 expression was detected by Western blot analysis in early virus infection.

RESULTSAY358935 was evolutionally conserved. The human AY358935 protein had an amino acid similarity of 74%, 60%, 38% and 33% with its counterpart in horses, mice, zebrafish and Xenopus laevis, respectively. Bioinformatics analysis indicated that AY358935 protein was located likely in the mitochondria. There was a N-terminal signal peptide and single transmembrane structure in AY358935 protein, which contained several phosphorylation sites. The secondary structure mainly comprised of alpha helices and random coils. AY358935 was ubiquitously expressed in normal tissues and carcinomas and regulated by the expression of double-stranded RNA-dependent protein kinase. AY358935 protein expression was obviously upregulated in cells 2 h after infection by vesicular stomatitis virus.

CONCLUSIONAs a predicted secretary protein with a small molecular weight, AY358935 might have important functions in cellular proliferation and anti-viral innate immune regulation.

Amino Acid Sequence ; Chromosomes, Human, Pair 11 ; genetics ; Computational Biology ; methods ; Humans ; Molecular Sequence Data ; Proteins ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Software ; Vesicular Stomatitis ; metabolism

OBJECTIVETo determine the infinite optical thickness of dentine porcelain of IPS E.max A color series.

METHODSCylindrical dentine porcelain specimens of the IPS E.max A color series were prepared with a diameter of 13 mm and thickness of 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, and 5.0 mm. The chromatic value of all the specimens was determined with CM-5 spectrometer against standard black and white background. The chromatic aberration (deltaE) was calculated by regression equation.

RESULTSThe infinite optical thickness of dentine porcelain of the IPS E.max A color series ranged from 2.341 to 3.333 mm for a deltaE of 1.0, and from 2.064 to 2.904 mm for a deltaE of 1.5. As the chromaticity or thickness increased, the influence by the background color decreased, and the color of specimens became gradually close to the intrinsic color.

CONCLUSIONThe thickness of the background dentine porcelain specimens must exceed its infinite optical thickness to represent the intrinsic color and avoid the influence by the extrinsic color.

Coated Materials, Biocompatible ; chemistry ; Crowns ; Dental Porcelain ; chemistry ; Dental Prosthesis Design ; Humans ; Prosthesis Coloring ; Tooth Preparation, Prosthodontic ; methods

ObjectiveTo establish the methodology for identification of clinical isolates of Mycobacterium and to identify the distribution of Mycobacterium species in hospitalized patients with tuberculosis in Heilongjiang province.It would provide the basis for accurate diagnosis of infections with Mycobacterium tuberculosis (MTB) and non-tuberculous Mycobacterium (NTM) as well as for effective therapy.Methods Three hundred and thirty Mycobacterium isolates from 330 tuberculosis patients hospitalized and clinically diagnosed in Harbin Chest Hospital from May 2007 to December 2008 were collected.Genomic DNA from the isolates was extracted after inactivation of Mycobacterium.Molecular identification was carried out using PCR,PCR-restriction fragment length polymorphism (RFLP) and sequencing.ResultsAmong 330 clinical isolates,328 were identified as MTB complex (MTBC),accounting for 99.4％ (328/330); 2 were NTM,accounting for 0.6％ (2/330).Among 328 MTBC isolates,326were MTB,one was Mycobacterium Africanum(M.African) and one was Mycobacterium microti(M,microti),accounting for 99.4％ (326/328),0.3％ (1/328) and 0.3％ (1/328),respectively.It was found that the homology between the two NTM isolates and Mycobacterium avium intracellulare (MAC)was 99％ and 93％,respectively,suggesting that the two NTM isolates were MAC.The homology between the two NTM isolates was 93％.ConclusionsPCR plus PCR-RFLP and sequencing provides an ideal method for precise identification of Mycobacterium species.In the clinically diagnosed tuberculosis patients,the predominant Mycobacterium species is MTB,however M.African,M.microti as well as NTM are also found.

Human-animal parasitic diseases caused by medical helminths are hazardous to human health. Genetic polymorphism studies on medical helminth populations can not only understand the biological characteristics and genetic structure of their populations, but also help reveal how they adapt to their parasitic environment, thus contributing to deepen our understanding of the epidemiological patterns of parasitic diseases and improve our understanding of accurate prevention and control of parasitic diseases. With the development of molecular biology, molecular markers such as DNA barcodes, simple sequence repeats, and single nucleotide polymorphism markers have been widely used to study the genetic relationships among parasite populations and individuals, and to reveal the genetic variation of parasite populations and the evolution of species origins. In this paper, we systematically review the application of three molecular markers commonly used in the study of genetic polymorphism in medical helminths, with a view to laying the foundation for related research.

OBJECTIVETo collect background information on drug-resistant HIV-1 strains in various regions before the start of nation-wide antiretroviral therapy in China.

METHODSTwenty percent of the 2,000 blood samples from antiretroviral therapy naive patients collected for the 2nd national HIV molecular epidemiology survey (NHMES) in 2002 were randomly sampled for this study. The entire protease gene and 20-230 amino acids of the reverse transcriptase gene were amplified by PCR from provirus DNA and sequenced. The results were analyzed with HIV db-Drug Resistance Algorithm and genotypic resistance mutations were determined to particular anti-HIV drugs.

RESULTSTotally 164 protease gene sequences and 138 reverse transcriptase gene sequences were obtained from patients; 0.61% of 164 sequences displayed primary resistance mutations in the protease gene, whereas 99.39% carried 1 or more secondary mutations. Genotypic resistance to at least one nucleoside reverse transcriptase inhibitors (NRTI) was present in 5.80%,and resistance to at least one non-nucleo side reverse transcriptase inhibitors (NNRTI) was present in 1.45% of samples.

CONCLUSIONThe prevalence of genotypic drug resistance is very low in drug-naive HIV infected patients from 21 provinces of China tested in this study. Laboratories participated in the NHMES have organized a network to provide drug resistance monitoring service in the current nation-wide antiviral treatment program in China.

Anti-HIV Agents ; therapeutic use ; China ; epidemiology ; Drug Resistance, Viral ; Genotype ; HIV Infections ; drug therapy ; epidemiology ; virology ; HIV Protease ; genetics ; HIV Protease Inhibitors ; therapeutic use ; HIV Reverse Transcriptase ; genetics ; HIV-1 ; genetics ; Humans ; Mutation ; Reverse Transcriptase Inhibitors ; therapeutic use ; Sentinel Surveillance

OBJECTIVETo investigate the effect of two methods of pigmentation on the flexural strength of dental Y-TZP/porcelain layered structure.

METHODSKaVo zirconia substructures were pigmented by dipping presintered blocks in the coloring solution VITA LL1 and LL5, and colored TZ-3YS zirconia substructures were fabricated by adding pigments before isostatic pressing. The colors No.1 and No.5 were used for the test. The specimens were made in monolithic or bilayered forms, and the flexural strength was tested. XRD and SEM with EDX were used to analyze the characteristics of the surface structure.

RESULTSIn KaVo group, no significant differences were found in the flexural strength between white and LL1 and LL5 colored monoclinic materials, nor in bilayered structures. While in TZ-3YS group, significant differences were noted in the flexural strength between color No.5 white and color No.1 monoclinic materials, but not between the latter two subgroups. The flexural strength was significantly lowered by veneering with porcelain in both zirconia groups, and similar findings were observed with the monoclinic materials. Only the tetragonal phase was detected in both of the zirconia groups.

CONCLUSIONPigmentation has no apparent effects on the bonding strength between the veneering porcelain and zirconia. Both coloring methods are appropriate when the concentration of the pigments is under deliberate control.

Dental Bonding ; Dental Materials ; Dental Porcelain ; chemistry ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Pigmentation ; Tensile Strength ; Yttrium ; chemistry ; Zirconium ; chemistry

OBJECTIVETo identify variations in the env gene of human immunodeficiency virus type 1 (HIV-1) subtype CRF01-AE strains circulating in China and to elucidate the potential relationship between genetic variation and evolutionary pressure.

METHODSFragments of the HIV-1 env gene were amplified by nested-polymerase chain reaction (n-PCR) from the whole blood of HIV-1 infected individuals from four provinces in Southeast China (Guangdong, Hunan, Jiangsu and Jiangxi). The PCR products were then directly sequenced by ABI 377 DNA sequencers. The sequences covering the env V3-V4 region of 34 HIV-1 subtype CRF01-AE strains were selected to analyse phylogenetic trees and amino acid mutations. The accumulation of synonymous (Ks) and antonymous (Ka) substitutions as well as Ks/Ka ratios were calculated using DIVERGE.

RESULTSPhylogenetic trees showed that the 34 HIV-1 subtype CRF01-AE strains from China clustered with the Chinese AE reference strain (AE.97CNGX2F), as well as with the reference strains from Thailand (AE.CM240 and AE.93TH253). The amino acid sequences of the env V4 and C3 regions in the samples were highly variable, compared with those of V3 and V3-downstream regions. The V3 loop central motif in the majority (87.5%) of the strains was GPGQ. The majority of strains did not contain positively charged amino acids at positions 306 and 320 in V3 loop. The N-linked glycosylation sites in the V3-V4 region and flanking regions in these strains were relatively conserved. Analysis of the entire region showed that the mean Ks values were significantly higher than that of the Ka values (P < 0.001), with the Ks/Ka significantly higher than 1.0 (P < 0.001). In contrast, the Ks/Ka ratio in the V4 region was significantly lower than 1.0 (P < 0.01).

CONCLUSIONSOur study indicated that the majority of HIV-1 subtype CRF01-AE strains circulating in China were highly homogeneous. The amino acid sequences of the V4 and C3 regions were significantly more variable than those of the V3 loop. Our analysis also suggested that the phenotype of nearly all strains was likely to be non-syncytium inducing (NSI). Finally, the variation found in the V3-V4 sequence was significantly influenced by functional constraints as opposed to positive selective pressure, while the variability of the lone V4 region was strongly related to positive selective pressure.

Adult ; Amino Acid Sequence ; China ; Evolution, Molecular ; Female ; Genes, env ; genetics ; Genetic Variation ; HIV-1 ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Sequence Homology, Amino Acid