Objective To investigate the situation and change of antifungal resistance in clinical Candida and other fungal iso- lates from 5 hospitals in diverse geographic region of China.Methods Antimicrobial susceptibility testing of 8 000 fungat iso- lates collected during 2001 and 2005 were carried out with 25?g fluconazole disk and 1?g voriconazole disk using disk diffusion method as recommend by CLSI/NCCLS M44-A.Disk test plates were automatically read and results were recoded with the BIOMIC Image Analysis System.The equivalent MICs were automatically calculated by the BIOMIC System software.Results The proportion of Candida atbicans and non-Candida albicans (e.g.Candida glabrata) in the total fungal isolates did not change significantly from 2001 to 2005.The susceptibility rate of C.albicans to fluconazole and vorieonazole were stable during 2001 and 2005.However, the resistance to fluconazole and voriconazole increased variably in C.glabrata and other non-Can- dida albicans fungal isolates during the same period.Conclusions The voriconazole demonstrated higher activity against all yeast species in comparison with fluconazole.The increasing resistance to fluconazole and voriconazole in non C.albicans fungal isolates including C.glabruta suggests the importance of surveillance of fungal resistance in Candida isolates.

Six DNA extraction methods and four DNA purification methods were compared and analyzed in this study to get higher quality DNA from the rhizospheric soil of Fritillaria thunbergii Miq.Results showed that higher purity DNA were harvested by pretreating the soil with 20 mmol/L EDTA(pH 7.5),then isolating soil DNA with CTAB-SDS-frozen-thawing,and further purified by agarose method.The recovery rate of this soil DNA was about 44.00 ?g/g ? 2.65 ?g/g soil,and they were qualified for the microbial diversity analysis in the rhizospheric soil of F.thunbergii Miq based on the 16S rDNA sequence.

OBJECTIVETo evaluate the clinical outcomes of percutaneous intervertebral foramina endoscopic lumbar discectomy for elder patients with lumbar spinal stenosis syndrome.

METHODSFrom July 2006 to July 2011, 60 elder patients with lumbar spinal stenosis syndrome were treated with surgical operation, including 32 males and 28 females with an average age of (66.7 +/- 2.5) years old ranging from 72 to 83 years. These patients were divided into the traditional surgery group and percutaneous intervertebral foramina endoscopic discectomy groups (PTED group), 30 cases in each group. The index of the preoperative and postoperative, operative incision visual analogue scale (VAS) of two groups were compared. The Oswestry disability index (ODI) of two groups at 6, 24 months of the follow-up were also evaluated on activity of daily living.

RESULTSThe average operative time, the average blood loss, the number of cases using analgesic drug, hospitalization time of PTED group were better than those of the traditional surgery group (P < 0.05). The improvement of incision VAS in PTED group was better than that in the traditional surgery group (P < 0.05). All patients were followed up for 24 months at least. The ODI at 1, 24 month after operation were better than that of preoperative in two group respectively (P < 0.05), but the improvement of PTED group was better than that of the traditional surgery group (P < 0.05).

CONCLUSIONPTED has the advantages of smaller incision, less bleeding, less postoperative stay and hospitalization time, tissue trauma and quicker recovery. It is a safe and efficacious minimally invasive surgical technique for elder patients with lumbar spinal stenosis syndrome.

Aged ; Aged, 80 and over ; Decompression, Surgical ; Diskectomy, Percutaneous ; Endoscopy ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Spinal Stenosis ; surgery ; Treatment Outcome

Bone Diseases ; therapy ; Bone and Bones ; physiology ; Humans ; Martial Arts ; education ; Medicine, Chinese Traditional ; methods ; Religious Philosophies

OBJECTIVETo determine the distribution of genotype IV among hepatitis E virus (HEV) infections in Wuhan by sequencing the open reading frame (ORF) 3 gene of HEV clinical isolates.

METHODSSerum samples were collected from 103 individuals who tested positive for the anti-HEV IgM antibody, and total HEV RNA was extracted for targeted gene sequencing analysis. Reverse transcription-nested polymerase chain reaction (PCR) was used to amplify two fragments of the ORF3 gene (5020 to 5392 nt and 5347 to 5956 nt, EF570133). The two PCR products were sequenced and the sequences were stitched with the ContigExpress program and used to determine the HEV genotype.

RESULTSBoth ORF3 gene fragments were amplified in 18 out of the 103 anti-HEV IgM-positive serum samples. These 18 HEV isolates shared 92.5% to 99.4% identity with each other at the nucleotide level. Nucleotide sequence homology analysis of the HEV genotypes I, II, III, and IV indicated the highest homology was with genotype IV; specifically, homology with genotype I was 83.5% to 86.7%, with genotype II was 83.2% to 85.2%, with genotype III was 84.6% to 87.2%, and with genotype IV was 92.0% to 96.5%.

CONCLUSIONTargeted sequencing of the HEV ORF3 gene facilitated genotyping of clinical isolates. Using this method, it was determined that nearly 20% of HEV clinical isolates from Wuhan belong to genotype IV.

Base Sequence ; Genotype ; Hepatitis E ; epidemiology ; virology ; Hepatitis E virus ; genetics ; Humans ; Open Reading Frames ; Sequence Homology, Nucleic Acid

Base Sequence ; Conjugation, Genetic ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Escherichia coli ; drug effects ; enzymology ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Polymerase Chain Reaction ; Transformation, Bacterial ; beta-Lactamases ; genetics

This study was purposed to investigate the intercellular cell adhesion molecule-1 (ICAM-1) gene K469E (A/G) (rs5498) and K56M (A/T) (rs5491) single nucleotide polymorphisms (SNP) and soluble ICAM-1 (sICAM-1) levels in plasma in three Chinese populations of Yugur, Tibetan and Han nationalities, to analyze comparatively the genotypes and allele frequencies distribution in different ethnic groups, and to explore the effects of ICAM-1 K469E and K56M polymorphism and sICAM-1 levels in plasma. EDTA-anticoagulant venous blood from Yugur(327 cases), Tibetan (400 cases) and Han (126 cases) people was collected, the DNA was extracted by using whole blood genomic DNA extraction kit, DNA SNP were analyzed by PCR-RFLP, genotype was judged by gel scan imaging system after agarose gel electrophoresis, the gene sequence was determined and the distribution of ICAM-1 genotypes and allele frequencies were compared among different ethnic groups, besides, the group representativeness was tested via the Hardy-Weinberg genetic equilibrium. Finally, the human sICAM-1 plasma levels were detected by using human ICAM-1 ELISA kit. The results showed that DNA sequencing result was consistent with PCR-RFLP analysis. In Yugur, Tibetan and Han nationalities, the KK, KE and EE three genotypes at ICAM-1 K469E gene locus were detected, the genotype distribution was not statistically significantly different, while the K, E allele frequency distribution was statistically significantly different (P < 0.05). Both of genotype and allele frequency distribution between Yugur, Tibetan and Han nationalities were statistically significantly different (P < 0.05). In K56M site only KK, KM two genotypes were detected, but the MM genotype was not detected in the three ethnic groups; the difference of two genotypes and K, M allele frequencies between Yugur and Han population was statistically significantly different (P < 0.05). Among three ethnic groups, the sex ratio and age distribution of K469E, K56M genotypes and allele frequencies of ICAM-1 gene were not significantly different, and distribution was in accordance with Hardy-Weinberg genetic equilibrium (P > 0.05). The plasma sICAM-1 level at ICAM-1 K469E allele locus in K individuals [(253 ± 122), (185 ± 97) µg/L] was higher than that at non-K allele [(145 ± 110) µg/L, P < 0.01]; the plasma sICAM-1 level of ICAM-1 K56M sites with KK genotype [(253 ± 122) µg/L] was higher than that of the KM genotypes [(168 ± 103) µg/L, P < 0.01]. In Yugur and Tibetan groups, the plasma sICAM-1 levels [(224 ± 80), (214 ± 111) µg/L] were higher than that in the Han group [(175 ± 125)µg/L, P < 0.05]. Pairwise comparison indicated that the plasma sICAM-1 levels between Yugur and Han group were statistically significantly different (P < 0.01), that was significantly different between Tibetan and Han group (P < 0.05). It is concluded that in Yugur, Tibetan and Han population, the genotypes and gene frequencies of two amino acid sites K469E and K56M in ICAM-1 were KK/KE-type, KK-type and K allele, moreover, the ratio of them in Yugur and Tibetan group was higher than that in Han, while there is not significant difference in sex ratio and age distribution, therefore, ICAM-1 genotype and allele frequency distribution in this study had ethnic representativeness. ICAM-1 gene K469E and K56M polymorphisms were likely to affect the plasma sICAM-1 expression level. K469E gene K allele may be a genetic risk factor, while K56M gene M allele a may be genetic protective factor for some diseases.
Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; genetics ; Male ; Middle Aged ; Plasma ; metabolism ; Polymorphism, Genetic ; Young Adult

The aim of this study was to investigate the similarities and differences of A1381T (rs216311) and -1793G/C (rs7966230) single nucleotide polymorphisms (SNP) in Chinese Yugur, Tibetan, and Han nationalities and their influence on plasma vWF concentration in order to explore the sensitivity of these 3 nationalities to vWF-related diseases. Peripheral venous blood was obtained from 322 Yugur, 399 Tibetan, and 120 Han healthy people. The DNA were then extracted. vWF gene A1381T and -1793G/C polymorphisms were analyzed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequenced when it was necessary. The vWF:Ag level in plasma was determined by ELISA. The results showed that the genotype distribution of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han nationalities was different with statistically significance (P < 0.05). GG genotype of A1381T locus accounted for 69.9% in Yugur nationality, which was much higher than 56.6% and 53.3% in Tibetan and Han nationalities respectively(P < 0.01); AA genotype of A1381T locus expressed a low level of vWF in plasma. For the -1793G/C locus, the proportion of CG genotype in Yugur was much higher than that in Han, CC genotype expressed a high level of vWF in plasma. The plasma vWF levels with different nationalities and the polymorphism of vWF gene were significantly different. It is concluded that the polymorphisms of vWF gene at both A1381T and -1793G/C loci in Yugur, Tibetan and Han are significantly different; the polymorphism of vWF gene influences the plasma vWF level; the plasma vWF levels in Yugur and Tibetan are significantly higher than that in Han, which may be associated with the living environment and habits.
Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Plasma ; chemistry ; Polymorphism, Genetic ; Young Adult ; von Willebrand Factor ; genetics ; metabolism

OBJECTIVETo study the distribution of cervical lymph nodes metastasis and explore the surgical treating modality of cervical lymph nodes in the patients with differentiated thyroid carcinoma.

METHODSThe clinic and pathological data of 104 patients with differentiated thyroid carcinoma who had undergone neck lymph nodes dissection from January 2003 to June 2007 were analyzed retrospectively. There were 29 male and 75 female patients. The age of the patients was 12 to 79 years old with a median of 39 years old. Patients were divided into clinic cervical lymph nodes metastasis (cN+) group and clinic no cervical lymph nodes metastasis (cN0) group according the condition of physical examination and image analysis preoperatively and compared respectively with pathological data postoperatively.

RESULTSIn the cN+ group 91.3% (63/69) patients were pN+ while in the cN0 group 52.1% (25/48) patients were pN+. The distribution of metastasized lymph nodes: level VI 64.1%, level II 31.6%, level III 44.4%, level IV 40.2%, level V 12.0%, level I 3.2%. In the cN+ group 86.7% (54/63) patients with lymph nodes metastasis had multi-levels lymph nodes metastasis while in the cN0 group 64.0% (16/25) patients had single-level lymph nodes metastasis.

CONCLUSIONSCervical lymph nodes metastasis in the patients with differentiated thyroid carcinoma mainly localize in level II, level III, level IV, level VI, especially level VI. Patients with lymph nodes metastasis had multi-levels lymph nodes metastasis in the cN+ group but single-level in the cN0 group. The surgical treating modality of cervical lymph nodes should also be different in the two group patients.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neck Dissection ; methods ; Retrospective Studies ; Thyroid Neoplasms ; pathology ; surgery

OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.

METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.

RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.

CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.

Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms ; therapy ; Receptors, Transferrin ; immunology ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; alpha-Fetoproteins ; genetics