Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; drug effects ; isolation & purification ; Anthracosis ; microbiology ; Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Fosfomycin ; pharmacology ; Humans ; Imipenem ; pharmacology ; Pneumonia, Bacterial ; microbiology

Objective To investigate the effect of isoflurane preconditioning on the nuclear factor kappa B (NF-kB)activity of leukocytes in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Twenty ASAⅡorⅢpatients of both sexes aged 20-60 yrs undergoing cardiac valve replacement under CPB were randomly divided into 2 groups(n=10 each):control group(C)and isoflurane preconditioning group(I).Anesthesia was induced with midazolam 0.08-0.12 mg?kg~(-1),fentanyl 5-10?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1).The patients were mechanically ventilated(FiO_2=100%)after tracheal intubation.Anesthesia was maintained with intermittent i.v.boluses of fentanyl and midazolam in group C or with 2 MAC isoflurane and intermittent i.v.boluses of fentanyl and midazolam in group I before CPB.Systolic BP was kept between 90-120 mm Hg,diastolic BP between 50-80 mm Hg and HR between 60-90 bpm in both groups. Isoflurane was discontinued at the initiation of CPB.Arterial blood samples were taken after tracheal intubation and before inhalation of isoflurane(T_0)at 30 min(T_1),1 h(T_2)and 2 h(T_3)after aortic unclamping for determination of NF-kB activity of leukocytes using electrophoretic mobility shift assay(EMSA).The amount of fentanyl,midazolam,dopamine and sodium nitroprusside(SNP)consumed during operation and the rate of recovery of spontaneous heart beat in both groups were recorded.Results The NF-kB activity was significantly increased after aortic unclamping in C group but there was no significant difference in NF-kB activity before CPB (T_0)and after aortic unclamping(T_(1-3))in I group.The NF-kB activity was significant lower at T_(1-3) in group I than in group C.The total amount of fentanyl consumed was 40-60?g?kg~(-1) in C group and 20-30?g?kg~(-1) in group I. Significantly less amount of dopamine was used in group I than in group C.There was no significant difference in SNP consumption between the 2 groups.The rate of recovery of spontaneous heart beat was significantly higher in group I than in group C(P＜0.01).The amount of dopamine consumed was positively correlated with the highest level of NF-kB activity in both group[r=0.962 in group C;r=0.908 in group I(P＜0.01)].Conclusion Isoflurane preconditioning can attenuate the NF-kB activity of leulocytes in patients undergoing cardiac valve replacement under CPB.

OBJECTIVETo explore the mechanism of the eye-acupuncture for treatment of acute cerebral ischemia-reperfusion injury.

METHODSThirty-two healthy SD rats were randomly divided into a normal group, a sham operation group, a model group and an eye-acupuncture group, 8 rats in each group. The rat model of cerebral ischemia-reperfusion was established with thread occlusion method in the model group and the eye-acupuncture group. The eye-acupuncture group was treated by eye-acupuncture at "liver region", "upper energizer area", "lower energizer area" and "kidney region" for 20 min immediately after reperfusion and at 30 min before sampling. No treatment was done in the normal group and the sham operation group, and no thread occlusion was performed in the sham operation group. The Neurologic impairment was scored and the methods of immunohistochemistry staining, western-blotting and real-time fluorescent quantitation polymerase chain reaction (RQ-PCR) were taken to detect the expression of the aquaporin protein 4 (AQP4) and its mRNA in cerebral cortex after reperfusion for 3 hours.

RESULTSThe neurologic impairment score of 1.50 +/- 0.54 in the eye-acupuncture group was significant lower than 2.63 +/- 0.92 in the model group (P < 0.01). The expression of the AQP4 protein by immunohistochemistry and western-blot respectively were 116.33 +/- 10.24 and 0.53 +/- 0.04 in the normal group, 118.97 +/- 12.72 and 0.55 +/- 0.07 in the sham operation group, and 129.30 +/- 18.36 and 0.67 +/- 0.08 in the eye-acupuncture group, with statistical significance compared to 150.88 +/- 15.82 and 0.94 +/- 0.04 in the model group (all P < 0.01), and there were significant differences between the eye-acupuncture group and the normal group (both P < 0.01). The tendency in the expression of AQP4 protein and its mRNA in all the group were almost the same.

CONCLUSIONThe eye-acupuncture therapy can relieve the cerebral ischemia-reperfusion injury and the protective mechanism is related to the downregulation of the cerebral AQP4 expression.

Acupuncture Therapy ; Animals ; Aquaporin 4 ; genetics ; metabolism ; Brain ; metabolism ; Brain Ischemia ; complications ; genetics ; metabolism ; surgery ; Disease Models, Animal ; Eye ; anatomy & histology ; Female ; Gene Expression ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; genetics ; metabolism ; therapy

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

OBJECTIVETo investigate on the expression of some cytokines and other immunity makers right after the operation, the effect of femoral nailing on systemic immunity and sought to differentiate any differences between reamed and unreamed IMN.

METHODSFifty-nine patients presenting with acute femoral fractured including 55 male and 4 female, 32.1 years old on average, are divided into 2 group depend on ISS. All patients were treated by close reduction and intramedullary nail for fixation. In group 1, 23 reamed and 23 unreamed; in group 2, 7 reamed and 6 unreamed. Venous blood samples were taken at 24 hr pre-operationally, and 1 hr, 24 hr, 48 hr post operationally. Serum TNF, IL-6, IL-8, IL-10 were measured by enzyme-linked immunosorbent assay. CRP was measured by protein assay apparatus. We also collected venous samples from 22 healthy uninjured volunteers, which formed control group.

RESULTSAll immune marks were elevated post operation, for IL-6, IL-8, IL-10, this elevation began at 1 hr after operation, reached to the peak at 24 hr, and then down but never to the normal at 48 hr. For TNF and CRP, the level were raised at 24 hr, and then fallen at 48 hr. All mediators were raised significantly above the control group (< 0.05). Between reamed and unreamed patients both in group 1 and group 2, Although there was a trend towards higher levels of TNF, IL-6, IL-8, IL-10 and CRP in RFN than in the URFN, no significant difference was found except that there was a greater release of serum IL-10 in RFN than in URFN at 24 hr post operation (P = 0.047). Two patients have become SIRS, but the markers have shown no significant difference with those that have no SIRS symptoms.

CONCLUSIONSTo the patient not injured severely, using IMN for treatment will make the inflammatory mediators re-released on higher level than normal, which will be balanced by immunity itself soon, so IMN won't make any damage severely. And no significant difference were found between reamed and unreamed nail. But the changing of IL-10 show us that after IMN, especially the reamed nailing, the level of anti-inflammatory mediators will show the difference more apparently between RFN and URFN while the patient got injured more severely. Under this condition, the RFN will aggravate the restrain of immunity.

Adult ; Biomarkers ; blood ; Female ; Femoral Fractures ; immunology ; surgery ; Fracture Fixation, Intramedullary ; adverse effects ; methods ; Fractures, Closed ; immunology ; surgery ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged ; Monitoring, Immunologic ; Postoperative Period ; Tumor Necrosis Factors ; metabolism

OBJECTIVETo explore the therapeutic effect of the modified maxillary bone disassembly procedures on patients with tumors in the anterior and middle skull base.

METHODSRetrospective analysis was made of patients treated with the modified maxillary bone disassembly procedures. Ten tumors in the anterior and middle skull base were resected according to the pathology, size and site of the skull base tumors including 9 benign tumors and 1 malignant tumor.

RESULTSAll tumors were resected completely. All patients were followed up from 12 months to 5 years postoperatively. The complications were less and the life quality was increased. The patients with benign tumors showed no recurrence . Two patients with cerebrospinal rhinorrhea cured spontaneously after 7 days. One patient with melanoma died of brain metastase.

CONCLUSIONSIt is necessary to estimate the tumors thoroughly before surgery. According to the location of the tumor, the modified maxillary bone disassembly is the nearest and harmless approach, through which the tumors can be completely excised with minimal invasiveness. The life quality is thus increased.

Adolescent ; Adult ; Female ; Humans ; Male ; Maxilla ; surgery ; Middle Aged ; Retrospective Studies ; Skull Base Neoplasms ; surgery ; Surgery, Oral ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the relationship between the resuscitation promoting role of resuscitation promoting factor and the initial bacteria amount of dormant Mycobacterium tuberculosis.

METHODSMycobacterium tuberculosis (dormant bacteria) was cultured for 100 days, then diluted into 1 mg/ml concentration with 7H9, and further diluted into 0.5, 0.25, 0.125, 0.0625, and 0.03125 mg/ml. Twelve new tubes added with 5 ml 7H9 and divided into two groups: the first group was added with the resuscitation-promoting factor protein, and the second group as control was added with 7H9. In each group the above diluted solutions were added. The tubes were located at 37 degrees C for culture. Optical density (OD) was detected on day 15, 25, 30, and 35. From each tube 1 microl culture solution was plated on 7H11 medium for colony counting.

RESULTSOD detection showed that bacteria proliferation in each group had positive linear correlation (P < 0.05, P < 0.01), indicating that the resuscitation-promoting factor played a similiar role in solutions with different dilution concentrations. 7H11 results and the OD results show that these two detection methods in each group had linear correlation (P < 0.05, P < 0.01), indicating that these two methods showed consistent test results.

CONCLUSIONThe resuscitation-promoting factor has no effect on the resuscitation of dormant Mycobacterium tuberculosis and its initial bacteria amount.

Bacterial Proteins ; metabolism ; Cytokines ; metabolism ; Mycobacterium tuberculosis ; physiology ; Resuscitation

OBJECTIVETo establish a rapid, inexpensive, and simple drug susceptibility test (DST) for Mycobacterium tuberculosis (M. tb) and evaluate its feasibility.

METHODWe used nitrate reductase combined with mycobacteriophage assay (PhaB-NRA) to test 49 clinical M. tb isolates of, and the results were compared with those of PhaB-NRA and traditional absolute concentration method.

RESULTSThe sensitivity, specificity, and accuracy of PhaB-NRA for rifampicin were 89.1%, 91.67%, and 89.8%; on the contrary, those of isonicotinyl hydrazide were 86.21%, 90.0%, and 87.8%, respectively. The coincidence between PhaB-NRA and traditional assay were 0.746 for rifampicin and 0.750 for isonicotinyl hydrazide.

CONCLUSIONSPhaB-NRA is an inexpensive, rapid, and simple DST method. It is a promising rapid screening technique for DST of M. tb.

Anti-Bacterial Agents ; pharmacology ; Biological Assay ; methods ; Humans ; Microbial Sensitivity Tests ; methods ; Mycobacteriophages ; physiology ; Mycobacterium tuberculosis ; drug effects ; Nitrate Reductase ; metabolism ; Rifampin ; pharmacology ; Sensitivity and Specificity

OBJECTIVETo investigate the drug resistance of imipenem-resistant (IR) Gram-negative bacilli (GNB) in coal workers' pneumoconiosis (CWP)-chronic obstructive pulmonary disease (COPD) patients with lower respiratory tract infection (LRTI) and to provide a basis for clinical treatment.

METHODSSixty-six strains of IR-GNB were isolated from the sputum of CWP-COPD patients with LRTI, and the bacterial spectrum was investigated. The drug resistance of bacterial strains was studied by KB disk diffusion method.

RESULTSAmong the 66 strains of IR-GNB, 29 (43.9%) were Pseudomonas aeruginosa, 17 (25.8%) were Acinetobacter baumannii, and 11 (16.7%) were Stenotrophomonas maltophilia. The drug sensitivity test showed that all bacteria had high drug resistance; Pseudomonas aeruginosa had a susceptibility rate higher than 50% to ciprofloxacin, polymyxin B, fosfomycin, and amikacin, Acinetobacter baumannii had a susceptibility rate higher than 55% to fosfomycin, polymyxin B, and cefoperazone/sulbactam, Stenotrophomonas maltophilia had a susceptibility rate higher than 50% to cotrimoxazole, ciprofloxacin, piperacillin/tazobactam, levofloxacin, polymyxin B, and cefoperazone/sulbactam, and Pseudomonas cepacia had a susceptibility rate higher than 50% to piperacillin/tazobactam, ciprofloxacin, cefoperazone/sulbactam, and polymyxin B.

CONCLUSIONThe main species of IR-GNB are such non-fermentative bacteria as Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia in CWP-COPD patients with LRTI. These bacteria have high drug resistance and are sensitive to only a limited range of antibiotics.

Aged ; Aged, 80 and over ; Anthracosis ; complications ; microbiology ; Drug Resistance, Multiple, Bacterial ; Gram-Negative Bacteria ; drug effects ; Humans ; Imipenem ; pharmacology ; Microbial Sensitivity Tests ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; complications ; microbiology

Eleven flavonol glycosides were isolated from the ethanol extract of Lysimachia clethroides by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were identified as astragalin (1), isoquercitrin (2), isorhamnetin-3-O-β-D-glucopyranoside (3), quercetin-3-O-β-D-6"-acetylglucopyranoside (4), quercetin-7-O-β-D-glucopyranoside (5), prunin (6), 2-hydroxynaringin-5-O-β-D-glucopyranoside (7), kaempferol-3-O-rutinonoside (8), kaempferol-3-O-robinobioside (9), rutin (10) and kaempferol-3,7-di-O-β-D-glucopyranoside (11). Among them, compounds 4, 7 and 11 were obtained from the Lysimachia genus for the first time, while compounds 3, 5 and 9 were firstly reported from this plant. In the preliminary assays, compounds 2, 6 and 8 possessed significant inhibition against aldose reduc- tase, with IC50 values of 2.69, 1.00, 1.80 μmol · L(-1), respectively; none of compounds 1-11 exhibited obvious cytotoxic activity (IC50 > 10 μmol · L(-1)).
Drugs, Chinese Herbal ; chemistry ; Flavonols ; chemistry ; Glycosides ; chemistry ; Molecular Structure ; Primulaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization