Animals ; Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Humans ; Interferons ; chemistry ; therapeutic use ; SARS Virus ; drug effects ; Severe Acute Respiratory Syndrome ; drug therapy ; virology ; Virus Diseases ; drug therapy ; virology

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; pathology ; Biopsy ; Biopsy, Fine-Needle ; Child ; Cytodiagnosis ; methods ; Diagnosis, Differential ; Female ; Humans ; Leukemia-Lymphoma, Adult T-Cell ; pathology ; Lymphoma, B-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Lymphoma, T-Cell ; pathology ; Male ; Middle Aged ; Pleural Effusion ; pathology ; Young Adult

PURPOSE: To evaluate the sealing performance of Hybrid Coat and its influence on the shear bond strength of five dentin surface cements. MATERIALS AND METHODS: Six premolars were pretreated to expose the dentin surface prior to the application of Hybrid Coat. The microscopic characteristics of the dentinal surfaces were examined with scanning electron microscopy (SEM). Then, 40 premolars were sectioned longitudinally, and 80 semi-sections were divided into a control group (untreated) and a study group (treated by Hybrid Coat). Alloy restoration was bonded to the teeth specimen using five different cements. Shear bond strength was measured by the universal testing machine. The fracture patterns and the adhesive interface were observed using astereomicroscope. RESULTS: SEM revealed that the lumens of dentinal tubules were completely occluded by Hybrid Coat. The Hybrid Coat significantly improved the shear bond strength of resin-modified glass ionomer cement (RMGIC) and resin cement (RC) but weakened the performance of zinc phosphate cement (ZPC), zinc polycarboxylate cement (ZPCC) and glass ionomer cement (GIC). CONCLUSION: Hybrid Coat is an effective dentinal tubule sealant, and therefore its combined use with resin or resin-modified glass ionomer cements can be applied for the prostheses attachment purpose.
Adhesives ; Alloys ; Bicuspid ; Dentin ; Glass Ionomer Cements ; In Vitro Techniques* ; Microscopy, Electron, Scanning ; Polycarboxylate Cement ; Prostheses and Implants ; Resin Cements ; Tooth ; Zinc Phosphate Cement

OBJECTIVETo observe the laryngopharynx manifestation and electromyography characteristics of myasthenia gravis (MG) patients.

METHODSThirty cases of MG were included in this study, their laryngopharynx symptoms and signs, voice acoustic assessment, laryngeal electromyography (LEMG) behaviors and repetitive nerve stimulation test(RNS) were analyzed, and the data was compared with that of normal subjects.

RESULTSAbout 36.7% of MG patients (11/30) had the symptoms of hoarseness, voice fatigue, dysphonia and dysphagia. The vocal folds movements of 16.7% of MG patients(5/30) appeared weaker than normal, and their vocal glottic couldn't close completely, while with a seam during phonation. Voice amplitude (68.3 +/- 14.6) dB (x +/- s, same at below), and maximum phonation time (15.1 +/- 4.0) s, were greatly lower than normal; shimmer(2. 43 +/- 1.19)%, and normalized noise energy (-9.6 +/- 3.3) dB, were greatly higher than normal. The amplitudes of interference patterns in MG patients' LEMG markedly decreased, except introarytenoid muscle, during low, normal and high pitch phonation, the amplitudes of thyoiarytenoid muscle were (215 +/- 69) microV, (298 +/- 113) microV and (380 +/- 153) microV, those of cricoarytenoid muscle were (253 +/- 92) microV, (361 +/- 116) microV and (486 +/- 155) RV. The turns increased but had no statistical difference. In the RNS test, 83.3% MG patients (25/30) showed masculine response. There were about 2.20 +/- 1.32 pieces of laryngeal muscles involved, and the reduction rate in amplitude of the compound muscle action potential for RNS was about (27.9 +/- 19.2)%.

CONCLUSIONSOnly parts of MG patients had laryngopharyngeal symptoms, but the laryngeal muscles of most of them were involved, appearing as the masculine response for RNS, the decreased synchronization of the laryngeal muscles' interference patterns, the decreased capacity of phonation. MG must be differentiated when a patient has the symptoms of voice weakness, hoarseness and dysphonia. Laryngeal RNS test should be used in the early diagnosis of MG.

Adolescent ; Adult ; Aged ; Articulation Disorders ; Case-Control Studies ; Electromyography ; Female ; Humans ; Hypopharynx ; physiopathology ; Laryngeal Muscles ; physiopathology ; Male ; Middle Aged ; Myasthenia Gravis ; physiopathology ; Young Adult

A simple and sensitive Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) method was established to provide a new alternative for clinical diagnosis of Avian influenza A H5N1 virus. The method employed a set of six specially designed primers that recognized eight distinct sequences of the target for amplification of nucleic acid under isothermal conditions. In current study, fifty-one experimentally infected animal specimens and viral cultures that had been tested were analyzed by RT-LAMP for NA gene and HA gene, respectively. The amplification process of LAMP was monitored in real-time by the addition of SYBR Green dye. Meanwhile, the result showed high correlation between nested PCR and RT-LAMP. The specificity of the RT-LAMP assay was confirmed by restriction enzyme digestion analysis and sequencing of the amplified product. When the sensitivity of this assay was tested by serial 10-fold dilutions of RNA molecules from specimens, it was found that the RT-LAMP method achieved theoretically a sensitivity of 10 RNA molecules. Thus, we concluded that the RT-LAMP assay has potential usefulness for rapid detection of the Avian influenza A H5N1 virus.
Animals ; Birds ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza in Birds ; diagnosis ; virology ; Nucleic Acid Amplification Techniques ; methods ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective The main objective of this study was to explore the prevalence and clinical characteristics of human coronavirus NL63 infection in hospitalized children with acute lower respiratory tract infection (ALRTI) in Changsha.Methods Nasopharyngeal aspirates (NPA) samples were collected from 1185 hospitalized children with ALRTI at the People's Hospital of Hunan province,between September 2008 and October 2010.Reverse transcriptase polymerase chain reaction (RT-PCR) was employed to screen for coronavirus NL63,which is a 255 bp fragment of a part of N gene.All positive amplification products were confirmed by sequencing and compared with those in GenBank.Results The overall frequency of coronavirus NL63 infection was 0.8％,6 (60％) out of the coronavirus NL63 positive patients were detected in summer,2 in autumn,1 in spring and winter,respectively.The patients were from 2 months to two and a half years old.The clinical diagnosis was bronchopneumonia ( 60％ ),bronchiolitis ( 30％ ),and acute laryngotracheal bronchitis( 10％ ).Four of the 10 cases had critical illness,4 cases had underlying diseases,and 7 cases had mixed infection with other viruses. The homogeneity of coronavirus NL63 with those published in the GenBank at nucleotide levels was 97％-100％.Conclusion Coronavirus NL63 infection exists in hospitalized children with acute lower respiratory tract infection in Changsha.Coronavirus NL63 infections are common in children under 3 years of age.There is significant difference in the infection rate between the boys and the girls:the boys had higher rate than the girls.The peak of prevalence of the coronavirus NL63 was in summer.A single genetic lineage of coronavirus NL63 was revealed in human subjects in Changsha.Coronavirus NL63 may also be one of the lower respiratory pathogen in China.

OBJECTIVETo investigate the clinical stage and histological grade of gastrointestinal stromal tumors.

METHODSTwelve clinical and pathological parameters were assessed in 613 patients with follow-up information. These parameters were classified into two gross spread parameters including liver metastasis and peritoneal dissemination, five microscopic spread parameters including lymph node metastasis, vascular, fat, nerve and mucosal infiltration, and five histological parameters including mitotic count ≥ 10 per 50 high-power fields, muscularis propria infiltration, coagulative necrosis, perivascular pattern and severe nuclear atypia.

RESULTSThe accumulated 5-year disease-free survival (DFS) and overall survival (OS) of 293 patients without any of these predictive parameters of malignancy were 99.3% and 100.0%, respectively. They were regarded as nonmalignant and further evaluations on the stage and grade of these tumors were not performed. At least one and at most seven predictive parameters of malignancy were identified in 320 patients. For these patients, the accumulated 5-year DFS and OS rates were 43.9% (mean 6.7 years) and 59.7% (mean 9.3 years), respectively. The DFS showed significant difference between patients with and without gross spread (P < 0.01), with and without microscopic spread (P = 0.001). DFS and OS were associated with the number of predictive parameters of malignancy in patients without gross spread (P < 0.01 for both DFS and OS), but not in patients with gross spread (P = 0.882 and 0.441, respectively).

CONCLUSIONSMalignant GIST could be divided into clinical stages I and II based on the absence and presence of gross spread, respectively. The degree of malignancy of patients in clinical stage I could be graded according to the number of predictive parameters of malignancy. Patients in clinical stage II were of the highest degree of malignancy regardless of the number of parameters. The staging and grading of gastrointestinal stromal tumors in this study are strongly associated with prognosis.

Actins ; metabolism ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; surgery ; Humans ; Liver Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; methods ; Neoplasm Invasiveness ; Neoplasm Staging ; methods ; Proto-Oncogene Proteins c-kit ; metabolism ; Survival Rate ; Young Adult

BACKGROUNDTo evaluate expression of human lactoferrin gene by high-density fermentation in recombinant Pichia pastoris on the premise of maintaining its biological activities.

METHODSThe neutrophil was isolated from human peripheral blood and its total RNA was prepared. Full-length cDNA of human lactoferrin gene was then obtained by RT-PCR, cloned into expression vector pPIC 3.5 K and transformed into Pichia pastoris strain KM71. With two-layer filter method, the transformants with high-productivity of human lactoferrin were screened out into fed-batch high-density fermentation. And later, the physical, chemical and biological activities of fermentation product were detected preliminarily.

RESULTSThe strain p3.5-k-7 with better productivity of human lactoferrin was screened out into fed-batch high-density fermentation. The fermentation lasted nearly for nine days, with A-600 of culture once above 260 and the highest productivity of human lactoferrin being 115 mg/L, 7.67 times the amount of that in shake flask cultivation.

CONCLUSIONThe authors successfully realized high-density fermentation expression of human lactoferrin gene in recombinant Pichia pastoris.

Cloning, Molecular ; Fermentation ; Humans ; Lactoferrin ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis

OBJECTIVETo investigate the impacts of interferon alpha-2b (IFN alpha-2b) on the oxidative stress states in the treatment of chronic hepatitis B (CHB) with different genotypes.

METHODSThirty-five patients with chronic hepatitis B and 18 healthy volunteers as a control were enrolled in this present study. In control and patients group, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), serum malondialdehyde (MDA) levels, serum total antioxidative stress capacity (TAC) were measured spectrophotometrically. After the therapy with interferon alpha-2b at the dose of 300 million units via intramuscular injection thrice a week for 12 weeks, these parameters were measured again in the patient group. The genotypes of hepatitis B virus were detected by polymerase chain reaction and hybridization. The effective group was defined as the patients with complete response and partial response.

RESULTSThe elevated concentrations of MDA and impaired levels of TAC in the patients with CHB were observed as compared to the healthy controls (P < 0.05 for both). There were no significant differences in serum levels of MDA and TAC in CHB patients with various genotypes (P > 0.05). The serum levels of MDA after the treatment with IFN alpha-2b were significantly lower than the pretreatment levels (P < 0.05), which even returned to the normal concentration (P > 0.05) in the effective group. There were significant increases in the TAC after the IFN alpha-2b therapy in the effective group. However, the significant differences in the TAC levels before and after the INFalpha-2b treatment were not observed in the non-responsive group.

CONCLUSIONThe oxidative stress could be improved with IFN alpha-2b treatment of chronic hepatitis B patients. The results suggest that antioxidant treatment for chronic hepatitis B patients may help improve the effect of anti-virus therapy.

Adolescent ; Adult ; Alanine Transaminase ; blood ; Antioxidants ; metabolism ; Antiviral Agents ; therapeutic use ; Aspartate Aminotransferases ; blood ; Female ; Genotype ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Malondialdehyde ; blood ; Middle Aged ; Oxidative Stress ; Recombinant Proteins ; Spectrophotometry ; Time Factors ; Treatment Outcome ; Young Adult

OBJECTIVETo construct a novel recombinant rhIFN-epsilon155ser, and study its biological activities.

METHODSThe whole sequence of rhIFN-epsilon was artificially synthesized and some codons were altered according to the preferred codon using of E.coli. The sequence was cloned into plasmid vector pBV220 to express in E.coli DH5alpha. After purification and re-folding of rhIFN-epsilon155ser inclusion body, the final product was tested for its biological activities, including anti-viral, anti-proliferative and NK cell enhancing activities. At the same time, by using DNA microarray biochips, the gene expression patterns in the rhIFN-epsilon155ser and rhIFN-alpha2b treated cells were compared and analyzed.

RESULTSThe re-built rhIFN-epsilon155ser sequence was expressed in E.coli as a form of inclusion body. After purified and re-folded, the rhIFN-epsilon155ser protein reached a purity of above 95%. The rhIFN-epsilon155ser protein had a specific anti-viral activity of about 6 x 10(5) IU/mg in WISH/VSV system. Its anti-proliferative activity and NK cell enhancing activities in vitro seemed to be lower than that of rhIFN-alpha2b. Data obtained from microarray biochips indicated that there were 283 pieces increasing 2 folds and 1489 pieces decreasing 2 folds among totally 22,278 pieces of human genes were found in the rhIFN-epsilon155ser treated cells; more changes in gene expression pattern were detected in the rhIFN-alpha treated cells.

CONCLUSIONA novel recombinant rhIFN-epsilon155ser was constructed, which belonged to type 1 interferon. The biological activities of rhIFN-epsilon155ser were compared with rhIFN-alpha2b. The changes of gene expression pattern in the interferon treated cells were detected, analyzed and discussed.

Antiviral Agents ; pharmacology ; Cell Proliferation ; drug effects ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; HeLa Cells ; Humans ; Interferons ; biosynthesis ; genetics ; pharmacology ; K562 Cells ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; Microbial Sensitivity Tests ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology