Aim In order to detect Mycoplasma fermentans (Mr)and Mycoplasma penetrans (Mpe) at the same time dual nested-PCR(DN-PCR) technique was established Method Mpe the same time PCR Primers were designed according to published 16s rRNA Sequences of Mf and Mpe, the outer primers were univevsal to both Mf and Mpe and the inner was Mf、Mpe speciespecific respectively. Two-step amplification procedures were perfomed. Result Chromo some DNAS of Mf and Mpe gave characteristic DN-PCR prafiles which were differeht from other my coplasmas or microorganism tested.Conclusion The DN-PCR method was established for the detection of Mf and Mpe at the same time which was sensitive specific and rapid.

OBJECTIVE To investigate mechanisms of aminoglycoside resistance in pan-drug resistant Pseudomonas aeruginosa(PDRPA).METHODS The 11 genes of 16S rRNA methylase and aminoglycoside-modifying enzymes were detected by polymerase chain reaction(PCR) and verified by DNA sequencing.RESULTS In 33 strains of PDRPA,the positive rates of rmtB,aac(3)-Ⅱ,aac(6′)-Ⅰb,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ were 42.4%,51.5%,42.4%,57.6%,48.5% and 63.6%.A total of 32 strains identified aminoglycoside modified genes,one strain aminoglycoside-modifying enzyme gene did not discovered,but rmtB positive.CONCLUSIONS The aminoglycoside resistance mechanisms of the PDRPA are the production of 16S rRNA methylase and aminoglycoside-modifying enzymes related.

OBJECTIVE To investigate mechanisms of drug resistance in multidrug-resistant strains of Acinetobacter baumannii(MDR-ABA).METHODS The resistant genes related to 5 kinds of antibiotics were detected by using polymerase chain reaction and verified by DNA sequencing.RESULTS In 30 strains of MDR-ABA,the positive rates of TEM,ADC,armA,aac(3)-Ⅰ,ant(3″)-Ⅰ,dfrA12 and sul1 were 63.3%,70.0%,30.0%,90.0%,90.0%,53.3% and 80.0%,respectively.CONCLUSIONS There are high positive percentages of TEM,ADC,armA,aac(3)-Ⅰ,ant(3″)-Ⅰ,dfrA12 and sul1 genes in MDR-ABA.

Objective To investigate the influence of postoperative cognitive dysfunction (POCD) to clinical outcome in adult patients and the risk factor. Methods 122 patients after operation on chest were surveyed with Wechsler Adult Intelligence Scale-Revised China (WAIS-RC) within 3 d before operation and on the 7th day after operation. Their pulmonary functions were measured before discharged. Their time on bed, total time in hospital, time after operation in hospital, time in intensive care unit and spending for therapy were investigated. Results 45 patients were finally diagnosed as POCD (POCD group) 7 d after operation. Before operation, the age, the body weight index, the smoking index and the number of risk factor of angiosclerosis in the POCD group were all higher than those whose cognitive function were normal (normal group) (P<0.05). The sores of insight, count, digit scope, vocabulary, figure collocation, figure patch of WAIS-RC in POCD group were obviously lower than those in normal group (P<0.05). Every index of pulmonary function in POCD group was lower than those in normal group(P<0.05). The time keeping on bed, total time in hospital, time after operation in hospital, time in intensive care unit and spending for therapy in POCD group all exceeded those in normal group (P<0.05).Conclusion The adult patients with POCD may spend more time and money for therapy in hospital, and their pulmonary functions may be recovered poorly.

Acupuncture Points ; Adolescent ; Adult ; Chronic Disease ; therapy ; Cough ; therapy ; Female ; Ginger ; chemistry ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

Aim To develop a molecular biologic technique for detection of chlamudia pneumoniae with nested polymeyase chain reaction (nPCR) .Methods Nested primers were synthesized according to a cloned C.pneumoniae 474- bp Pst I fragmert. Results The 378bp DNA fragments were amplified from C. pneuomoniae with nPCR. None of the C. trachomatis, C. psittaci, other organisms and etc. Strains tested were amplified by the nPCR. The direct sequening of 3 sample products with nPCR are quite same as C. pneumoniae (CWL-29 ) . The sensitivity of nPCR is higter than PCR. Conclusions This method is not only sensitive.specific and rapid, but also provides an etiological basis for cdiagnosis of C.pneumoniae infection.

Objective　To investigate the effects of trace elements on the metabolism of extracellular matrix and explore the physiological and pathological mechanism of trauma. Methods　Based on the experimental and clinical data, it was studied that the action of trace elements in the metabolism of extracellular matrix in trauma repairing. Results　During wound healing, the trace elements were the components of many kinds of enzymes, carriers and proteins. They took part in the synthesis of hormones and vitamins as well as the transmission of information system. They activated many different kinds of enzymes and regulate the levels of free radicals. The trace elements had the complicated effects on the synthesis, decompose, deposition and reconstruction of collagen and other extracellular matrix. Conclusion　The trace elements play an important role in regulating the metabolism of extracellular matrix.

Objective To analyze molecular evolution and binding free energies in substrates of KPC-2,KPC-5 and KPC-10 carbapenemases.Methods Minimum Evolution method in MEGA 4.1 was used to analyze molecular evolution of KPC-2,KPC-5 and KPC-10 carbapenemases,Dock module in ArgusLab 4.1 was used to perform molecular docking of these 3 carbapenemases to 10 kinds of β-lactams substrates,and calculate binding free energies(△G).Results Ambler Class A β-lactamases with carbapenemase activities were grouped in the same cluster and had good conservation,while ordinary Ambler Class A β-lactamases without carbapenemase activities were groupod in the other cluster.Binding free energies of KPC-2,KPC-5 and KPC-10 carbapenemases were lower to carbapenem antibiotics than the thirdgeneration cephalosporins,while binding free energies to aztreonam and clavulanic acid were of comparatively higher levels.Conclusion Catalytic activities of KPC to carbapenem antibiotics are higher than those to the third-generation cephalosporins,but the activities to aztreonam and clavulanic acid are low.

Aim To investigate the protective effects of etomidate on cultured rat hippocampal neurons subjected to anoxia injury and its mechanism.Methods Hippocampal neurons of neonatal rat,which had been cultured in vitro for 10 days,were allocated to control groups and etomidate-treating groups.The neurons were exposed to oxygen-glucose deprivation for 24 h.The cell survival rate in each group was evaluated using MTT colorimetry.To explore the effect of etomidate on neuronal calcium overload evoked by anoxia or 50 mmol?L~(-1) KCl or 1 mmol?L~(-1) glutamate,fluo-3,a fluorescent probe,was used for imaging of intracellular calcium in laser scanning confocal microscope(LSCM)to measure real-time changes of [Ca~(2+)]_i in cultured rat hippocampal neurons.Results The hippocampal neurons developed acute neuronal swelling and widespread neuronal degeneration following anoxia for 24 h.Etomidate at concentrations of 1.2～4.8 mg?L~(-1) attenuated the neuronal injury in a dose-dependent manner(P

Objective To investigate the effect of propofol on N-methyl-D-aspartate (NMDA) receptor-mediated currents. Methods Hippocampal neurons were prepared from newborn Wistar rats and cultured for 8-12 days. Whole cell currents were recorded using patch-clamp technique and cells were voltaged-clamped at - 80 mV. 100 ?mol?L-1 NMDA and 3 or 48 ?g?ml-1 propofol were applied with a multi-pipe ejection system. GABAA receptor was then blocked with 100 ?mol?L-1 bicuculline to investigate the effect of propofol on NMDA receptor without the influence of GABAA receptor.Results Run-down of INMDA induced by 100 ?mol?L-1 NMDA applied to neurons which were cultured for 8-12 days was 15%?8%. Propofol 3 or 48 ?g?ml-1 significantly inhibited spontaneous excitatory postsynaptic currents and elicited a Cl- -mediated response by direct activation of GABAA receptor in the absence of GABA. therefore produced a reversible inhibition of whole cell currents activated by NMDA. After the GABAA receptor was blocked by 100 ?mol?L-1 bicuculine propofol still inhibited NMDA currents slightly. Conclusion Propofol inhibits the NMDA subtype of glutamate receptor mainly through activation of GABAA receptor. It can also directly suppress the NMDA receptor slightly.