OBJECTIVETo clone pIA and pIB genes of Neisseria gonorrhoeae,and to construct pIA-pIB fusion gene and its prokaryotic expression system, and to establish enzyme linked immunosorbent assay (ELISA) based on rPIA-PIB for detecting serum and pus samples from gonorrhea patients and to evaluate the sensitivity and specificity of the ELISA.

METHODSpIA-pIB fusion gene was constructed by polymerase chain reaction (PCR) using linking primers and a prokaryotic expression system of the fusion gene was constructed by using routine molecular biological methods. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) plus BioRad Gel Image Analyzer was used to measure the expression of the target recombinant protein rPIA-PIB. Ni-NTA affinity chromatography was performed to extract and purify rPIA-PIB. An ELISA by using rPIA-PIB as the coated antigen for detecting the specific IgG against rPIA and/or rPIB in gonorrhea patients' sera as well as another ELISA by using rPIA-PIB antiserum as the first antibody for detecting the rPIA and/or rPIB in gonorrhea patients' pus samples were established. In these experiments, ELISAs associated with rPIA, rPIB and their antisera were applied as the controls.

RESULTS100% similarities of the nucleotide and putative amino acid sequences of the pIA-pIB fusion gene were confirmed when compared with the original sequences. The output of rPIA-PIB was 29.8% of the total bacterial proteins. The purified rPIA-PIB only showed a single target protein segment in gel after SDS-PAGE. Using a positive rate (98.3%) of rPIA-PIB-IgG-ELISA to detect 119 cases of gonorrhea patients' serum samples was remarkably higher than that of rPIA-IgG-ELISA (30.3%) or rPIB-IgG-ELISA (66.4%) (P<0.01). The positive rate (91.6%) of rPIA-PIB-ELISA to detect 119 cases of gonorrhea patients' pus samples was also significantly higher than that of rPIA-IgG-ELISA (27.7%) or rPIB-IgG- ELISA (62.2%) (P<0.01).

CONCLUSIONIn this study we successfully constructed pIA-pIB fusion gene of N. gonorrhoeae and its prokaryotic expression system while rPIA-PIB showed obvious superiority used as the antigen in gonorrhea associated detection kits compared to both the rPIA and rPIB.

Antigens, Bacterial ; immunology ; Bacterial Outer Membrane Proteins ; genetics ; immunology ; metabolism ; Base Sequence ; Enzyme-Linked Immunosorbent Assay ; methods ; Gene Expression Regulation, Bacterial ; Humans ; Neisseria gonorrhoeae ; genetics ; immunology ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo assess the clinical efficacy of acupuncture and moxibustion on depression in view of the outcome indicators of the patient subjective reports.

METHODSOne hundred and sixty-three cases of depression being in compliance with the inclusive standards were randomized into a soothing-liver and regulating-mind group, an acupoint-shallow-puncturing group and a non-acupoint-shallow-puncturing group. In the soothing-liver and regulating-mind group, the conventional acupuncture was applied to the four-gate points [Hegu (LI 4) and Taichong (LR 3)], Baihui (GV 20) and Yintang (EX-HN 3), the direct moxibustion with moxa cone was applied to the four-flower points [Geshu (BL 17), Danshu (BL 19)]. Finally, the intradermal needling was used at Xinshu (BL 15) and Ganshu (BL 18). In the acupoint-shallow-puncturing group, the acupoints selected were same as those in the soothing-liver and regulating-mind group. But the needle insertion was shallower and the time of moxibustion was shorter. In the non-acupoint-shallow-puncturing group, the spots that were 10 mm lateral to those acupoints in the soothing-liver and regulating-mind group were selected. The operation was same as that in the acupoint-shallow-puncturing group. The treatment was given twice a week in three groups. Totally, 12 weeks of treatment were required. The score of symptom checklist 90 (SCL-90), the self-report symptom inventory was observed before treatment, 1 month and 3 months after treatment separately so as to assess the corresponding short-term, mid-term and long-term efficacies of the program of acupuncture and moxibustion for soothing the liver and regulating the mind.

RESULTSIn each time-point after treatment, for the scores of somatization, obsessive-compulsive symptom, interpersonal sensitivity, depression, anxiety, hostility, paranoid ideation, psychoticism and the other 8 dimensionalities, in comparison between the soothing-liver and regulating-mind group and the non-acupoint-shallow-puncturing group, the differences were significant statistically (all P < 0.05). For the scores of depression, anxiety and hostility, in comparison between the soothing-liver and regulating-mind group and the acupoint-shallow-puncturing group, the differences were significant statistically (all P < 0.05).

CONCLUSIONAcupuncture and moxibustion can improve the scores of SCL-90 scale for the patients with depression. The outcome indicators of the patient subjective reports can accurately assess the clinical efficacy.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Depression ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Self-Assessment ; Treatment Outcome ; Young Adult

AIMTo establish a solid phase extraction-high performance liquid chromatographic (SPE-HPLC) method for determining plasma scutellarin concentration, and study its pharmacokinetics after i.v. breviscapine in rabbits.

METHODSMethanol-water-phosphoric acid (50:50:0.5) mixture was used as mobile phase, Nucleosil C18 column (250 mm x 4.6 mm ID) was selected. The wavelength of UV detection was 335 nm. Fifteen rabbits, randomized into 3 groups, were given breviscapine i.v. at the dose of 10, 20 and 40 mg.kg-1. Scutellarin in plasma was determined by SPE-HPLC method.

RESULTSLinearity was obtained over the range of 0.02-10.0 mg.L-1 of scutellarin. The method recovery was 96.15%-99.31%; the within-day and between-day RSDs were all below 10%. After i.v. 10, 20 and 40 mg.kg-1 of breviscapine to rabbits, the concentration-time curve of scutellarin fitted to a three compartment model. The main pharmacokinetic parameters showed no significant difference between low and medium doses, but the difference was significant between high dose and other doses.

CONCLUSIONThis assay method was accurate, sensitive, simple and suitable for the measurement of plasma scutellarin concentration. The pharmacokinetic characteristics were found to fit a three-compartment model following i.v. injection of breviscapine to rabbits. The changes of drug concentration in vivo exhibited linear kinetics ove the dosage range of 10-20 mg.kg-1, but when the dosage was 40 mg.kg-1, the linear kinetic properties disappeared.

Animals ; Apigenin ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Flavonoids ; blood ; pharmacokinetics ; Male ; Rabbits

Objective To investigate the clinical effects and treatment of thyroid dysfunction induced by PegIFNα-2a therapy in patients with chronic hepatitis B.Methods 95 patients with chronic hepatitis B received PegIFNα-2a from 2007-2011were selected. Analyzed the clinical manifestations,laboratory results,treatment and clinical outcome of the patients who had thyroid dysfunction induced by PegIFNα-2a therapy.Results During the treatment of PegIFNα-2a,10 patients ( 10/95,10.53％ ) had thyroid dysfunction,in which 4 were of hyperthyroidism and 6 were of hypothyroidism.After appropriate treatment,all the patients who had thyroid dysfunction had satisfactory outcome.Conclusions PegIFNα-2a therapy may induce thyroid dysfunction.Monitor thyroid function regular and have active treatment ensure the interferon therapy to continue.With the end of the interferon therapy,thyroid function can be back to normal.

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

This paper presents a novel monitor which uses ARM controller AT91SAM7S64 as its main processor, LCM (Liquid Crystal Display Module) for displaying ECG waves, SD (Secure Digital memory) card for data storage and RF module PTR8000 for radio data transmission. This portable monitor boasts alarm function for abnormality and can provide dynamic ECG monitoring for patients.
Computer Communication Networks ; Electrocardiography ; instrumentation ; Humans ; Monitoring, Physiologic ; instrumentation ; Signal Processing, Computer-Assisted ; instrumentation ; Telemetry ; instrumentation ; methods

OBJECTIVETo detect the specific mutations in rpoB gene of Mycobacterium tuberculosis by oligonucleotide microarray.

METHODSFour wild-type and 8 mutant probes were used to detect rifampin resistant strains. Target DNA of M. tuberculosis was amplified by PCR, hybridized and scanned. Direct sequencing was performed to verify the results of oligonucleotide microarray.

RESULTSOf the 102 rifampin-resistant strains 98 (96.1%) had mutations in the rpoB genes.

CONCLUSIONOligonucleotide microarray with mutation-specific probes is a reliable and useful tool for the rapid and accurate diagnosis of rifampin resistance in M. tuberculosis isolates.

Antibiotics, Antitubercular ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; DNA-Directed RNA Polymerases ; Drug Resistance, Bacterial ; genetics ; Gene Expression Regulation, Bacterial ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; metabolism ; Oligonucleotide Array Sequence Analysis ; Rifampin ; pharmacology

Influenza is a global contagious disease, which causes hundreds of thousands of people deaths. And new subtype of influenza may cause pandemic influenza, and lead to more serious consequence. Drugs for anti-influenza have played very important roles in influenza treatment and prevention, especially neuraminidase inhibitors are effective on both influenza A and B which have more safety and tolerance, therefore they have been widely used for influenza treatment. However, several viral drug-resistant cases have been reported. In this paper, the clinic therapy, prevention and drug resistance of neuraminidase inhibitors, including zanamivir, oseltamivir and peramivir, and progress in the research and development in our country are presented in order to promote research and development of new drugs for influenza treatment and prevention.
Antiviral Agents ; pharmacology ; therapeutic use ; Drug Resistance, Viral ; drug effects ; Enzyme Inhibitors ; pharmacology ; therapeutic use ; Humans ; Influenza, Human ; drug therapy ; prevention & control ; virology ; Neuraminidase ; antagonists & inhibitors ; Oseltamivir ; pharmacology ; therapeutic use ; Zanamivir ; pharmacology ; therapeutic use

Adult ; Aged ; Female ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Multiple Trauma ; complications ; nursing ; surgery ; Orthopedic Procedures ; Shock ; nursing

OBJECTIVETo examine mitochondrial DNA mutations in mitochondrial myopathy.

METHODSThree suspected cases of mitochondrial myopathy were examined by HE staining, histochemical staining methods and electron microscopy. The mutations in all 22 tRNA genes of mitochondrial genome were screened by polymerase chain reaction-single strand conformation polymorphism and DNA sequencing.

RESULTSThe three cases were diagnosed as mitochondrial myopathy. The examinations revealed that patient 1 had a homoplasmic A1627G mutation in tRNA-Val gene, and patient 2 had a heteroplasmic A1627G/A mutation in tRNA-Val gene, and patient 3 had two mutationsuone was homoplasmic T5554C mutation in tRNA-Trp gene, the other was heteroplasmic A10412C/A mutation in tRNA-Arg gene.

CONCLUSIONtRNA genes mutations of mtDNA might be one of the etiologies of mitochondrial myopathy.

Adult ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Female ; Humans ; Male ; Microscopy, Electron, Transmission ; Mitochondrial Myopathies ; genetics ; pathology ; Muscle Fibers, Skeletal ; metabolism ; pathology ; ultrastructure ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; RNA, Transfer, Val ; genetics ; Young Adult