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

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 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

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 compare the application of HE and enzyme histochemical staining in assessing the viability of hepatocellular carcinoma (HCC) cells coagulated by microwave ablation at different temperatures.

METHODSTwo groups of mice (n=6) with transplanted homogenic HCC were treated by microwave ablation at 60 degrees C and 50 degrees C for 3 min, respectively. Before and after microwave ablation, paraffin sections and frozen sections of the tumors were prepared for routine HE staining and enzyme histochemical staining with nicotinamide adenine dinucleotide diaphorase (NADH-diaphorase), respectively, and observed under microscope.

RESULTSShortly after microwave ablation, the morphology and arrangements of the nucleus of the ablated tumor cells in the two groups showed no obvious alteration in HE stained sections, but in sections with enzyme histochemical staining, the activity of NADH-diaphorase in ablated tumor tissue at 60 degrees C disappeared, suggesting the death of HCC cells; sporadic activity of the enzyme was detected in the coagulated tumor at 50 degrees C, indicating tumor cells surviving the ablation. The ablation effect was markedly different between the two groups (P<0.01).

CONCLUSIONHE staining is not suitable for evaluation of HCC destruction immediately after microwave ablation, and detection of NADH-diaphorase activity with the enzyme histochemical method better suits this purpose.

Animals ; Catheter Ablation ; methods ; Dihydrolipoamide Dehydrogenase ; metabolism ; Female ; Histocytochemistry ; methods ; Liver Neoplasms ; enzymology ; pathology ; therapy ; Liver Neoplasms, Experimental ; enzymology ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Microwaves ; therapeutic use ; Temperature

OBJECTIVETo study the clinical effects and security of nanometer ceramics artificial bone transplantation to treat the bone defect.

METHODSFrom March 2005 to November 2007, 32 patients (artificial bone group) with extremity bone defects applied nanometer ceramics artificial bone transplantations, included 19 males and 13 females, aged from 17 to 63 years old (averaged 31.4 years). The other 36 patients (internal fixation group) with extremity bone defects were treated by the internal fixation in the same period, included 21 males and 15 females, aged from 16 to 65 years old (averaged 32.6 years). Ca, P, B-ALP, IgG, IgA, IgM, CIC, C3, SL-2R and CD4+/CD8+ in the peripheral venous blood were measured in the 1st and 2th week and 1st, 3rd, 6th month after operation. All patients were followed up and the limb function was evaluated according to Enneking standard.

RESULTSThe wounds of all patients smoothly healed after operation. Every immunological indicators had no significant difference between two groups. Serum calcium and phosphorus content did not significantly increased. Serum B-ALP of all patients were increased after operation, fell to normal levels in the internal fixation group, but remained at a relatively high level in the artificial bone group. All patients were followed-up for from 9 to 24 months (averaged 15 months). All patients get the excellent physical function.

CONCLUSIONThe artificial bone has no immunogenicity, no rejection,does not affect the blood calcium and phosphorus content, and has higher osteogenic activity. It is affirmed that nanometer ceramics artificial bone is used to treat the smaller bone defect on clinical.

Adolescent ; Adult ; Bone Transplantation ; immunology ; Bone and Bones ; immunology ; metabolism ; pathology ; Calcium Phosphates ; chemistry ; Ceramics ; Female ; Follow-Up Studies ; Graft Rejection ; Humans ; Male ; Middle Aged ; Nanoparticles ; Prostheses and Implants ; Young Adult

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

In order to investigate the effects of phenylhexyl isothiocyanate (PHI) on proliferation and apoptosis of multiple myeloma cells RPMI8226 in vitro, the RMPI8226 cells were co-cultured with PHI at various concentrations; the apoptosis of PHI-treated cells was assayed by TUNEL; the cell cycle changes of PHI-treated cells were analyzed by FCM; the mitochondrial potential changes of PHI-treated cells were detected by using a potential sensitive dye JC-1 as probe; the VEGF levels secreted from PHI-treated cells were measured by quantitative sandwich ELISA. The results showed that PHI significantly inhibited RPMI8226 cell proliferation, induced their apoptosis at low concentration (0.5 micromol/L), the inhibitory effect was related to PHI concentration. PHI-treated cells were arrested in G(0)/G(1) phase. The RPMI8226 cells showed shift from red fluorescence to green fluorescence in some concentration-dependent manner, indicating increase of mitochondrial depolarization and potential loss by 3-4-fold as compared with control, after treated RMPI8226 cells with 10 micromol/L of PHI for 48 hours, the VEGF level secreted from RMPI8226 cells significantly decreased, it was 35% of control. It is concluded that the PHI can inhibit cell proliferation, induce cell apoptosis of RMPI8226, the cell apoptosis is associated with mitochondrial depolarization and potential loss, the inhibiting VEGF secretion from RMPI8226 cells by PHI may be one of the reasons causing apoptosis. PHI may be a potential therapeutic drug for multiple myeloma.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Isothiocyanates ; pharmacology ; Multiple Myeloma ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; secretion