Objective To investigate the role of cyclooxygenases (COXs) in the up-regulation of the expression of P2X3 receptors in the dorsal root ganglion (DRG) in rats with neuropsthic pain. Methods Twenty-four male SD rats, weighing 250-280 g, were randomly divided into 4 groups ( n = 6 each): sham operation group (group S), chronic constrictive injury (CCI) group, COX-1 inhibitor ibuprofen group (group Ⅰ), and COX-2 inhibitor celecoxib group (group C). Neuropathic pain was induced by CCI. The animals were anesthetized with intraperitoneal 10% chloral hydrate 300-500 mg/kg. CCI was produced by placing 4 ligatures on the left sciatic nerve at 1 mm intervals. In group S, the left sciatic nerve was only exposed but not ligated. In groups Ⅰ and C, ibuprofen 40 mg·kg-1 ·d-1 and celecoxib 30 mg·kg-1 ·d-1 were given through a gastric tube into the stomach at day 3-14 after operation respectively. Paw withdrawal latency (PWL) and paw withdrawal threshold (PWT) were measured before operation (baseline), and at 3, 5, 7, 10 and 14 days after operation. Then the rats were sacrificed and their L()-6 DRGs were removed to detect the expression of P2X3 mRNA and protein. Results Compared with group S, PWL was significantly shortened, PWT decreased, and P2X3 mRNA and protein expression up-regulated in group CCI ( P ＜ 0.05＝. Compared with group CCI, PWL was significantly prolonged, PWT increased, and P2X3 mRNA and protein expression down-regulated in groups Ⅰ and C (P ＜0.05＝. Compared with group Ⅰ, PWL was significantly prolonged, PWT increased, and P2X3 mRNA and protein expression up-regulated in group C ( P ＜0.05＝. Conclusion COXs are involved in the up-regulation of the expression of P2X3 receptors in the DRG in rats with neuropathic pain, and the effect of COX-1 is stronger than that of COX-2.

Objective To study the clinical characteristics,antimicrobial restistance of bloodstream infections (bacteremia) caused by multidrug-resistant Acinetobacter and analyze the outcomes of antibacterial therapy.Methods The clinical data were reviewed retrospectively for 74 patients with bloodstream infection caused by multidrug-resistant Acinetobacter who were trea-ted in HuaShan hospital from January 2005 to December 2011 .Results During the 6-year period,74 patients were diagnosed with multidrug-resistant Acinetobacter bacteremia,73 of which were nosocomial infections.The remaining one was community-acquired. Primary bloodstream infection accounted for 51 .4% (38/74),and secondary infection 48.6% (36/74), mainly secondary to pulmonary infections (23.0%,17/74). Solid tumor was the most common underlying disease (24.3%,18/74).Prior corticosteroid therapy,indwelling deep venous catheter,surgery and invasive procedures were predisposing factors of bacteremia. Acinetobacter-related bloodstream infections were associated with higher white blood cell count,increased neutrophil percentage,higher APACHE II score and lower serum albumin level.The bloodstream infection was caused by Acinetobacter baumannii in 65 pa-tients,Acinetobacter lwoffi in 7 patients,both Acinetobacter baumannii and Acinetobacter junii in one patient.The all-cause mortality rate was 27.0% (20/74).In vitro susceptibility testing showed that 20.0% (15/75 )of the Acinetobacter isolates were resistant to cefoperazone-sulbactam,which was the lowest among all the antibiotics tested.About 40.0% to 42.7% of the isolates were resistant to carbapenems.The outcome was related to the antimicrobial restistance.Carbapenem non-suscepti-ble Acinetobacter was associated with poorer outcome compared with carbapenem-susceptible Acinetobacter (mortality 46.9%vs 11 .9%,P <0.05 ).Cefoperazone-sulbactam non-susceptible Acinetobacter was also associated with poorer outcome com-pared with cefoperazone-sulbactam susceptible Acinetobacter (mortality 40.0% vs 18.2%,P <0.05).Of the 32 patients who had infections with carbapenem-non-susceptible Acinetobacter,20 received sulbactam-containing antimicrobial agent.The mor-tality of these 20 patients was 20.0% (4/20),significantly lower than that of the 12 patients who did not receive sulbactam-containing antimicrobial agent (66.7%).Conclusions Majority of the bloodstream infections caused by multidrug-resistant Acinetobacter are nosocomial infections.Surgical operation and serious condition may predispose the patients to develop Acine-tobacter bacteremia.Acinetobacter isolates are highly resistant to commonly used antibiotics.The Acinetobacter isolates not susceptible to carbapenem or cefoperazone-sulbactam are associated with poorer outcome and higher mortality.More attention should be paid to prevention and control of Acinetobacter-related nosocomial infections.

Objective To research the antiviral activity of artesunate (ART) in vitro fighting against both standard laboratory strains and ganciclovir(GCV)-resistance strains of human cytomegalovims(HCMV) and to explore whether fractionation dosage method can obviously enhance the antiviral effect of ART.Methods 1.Cytotoxicity assay to ART was performed by the use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetry.The 0％ toxic concentration (TC0) were determined,and median cytotoxic concentration (TC50) was calculated with Probit regression method.2.Antiviral activity assays of ART against HCMV:human embryonic lung fibroblast cells (HELs) were infected with standard laboratory strains and GCV-resistance strains of HCMV,respectively,after which virus was removed and overlays of dulbecco's modified eagle medium(MEM) containing different antiviral drugs were added to the wells.All cells were cultured continuously at 37 ℃ in a 50 mL/L CO2 humidified atmosphere for 7-10 days and the cytopathic effect (CPE) was observed under a microscope.When the degree of CPE was clear (+ + +-+ + + +),the values of absorbency at 490 nm of all cell wells were measured by MTT colorimetry.The cell survival rate (CSR)and drug inhibitory rate (IR) for HCMV were calculated.By Probit regression method,the median inhibitory concentration (IC50) of 2 drugs was calculated respectively.3.To explore whether fractionation dosage method could obviously enhance the antiviral effect of ART against HCMV,the experiment was divided into 3 groups and compared with GCV group,respectively:Group 1:ART antiviral compounds were added to cell layers by one dosage.Group 2:Total drug dosage was divided into 3 parts,and each part was added to cell layers once a day for 3 days.Group 3:Total antiviral compounds were divided into 6 and delivery 2 times a day.The values of absorbency at 490 nm of all cell wells were measured by MTT colorimetry.The CSR and viral inhibitory rates were calculated.All data were statistically analyzed by One-Way ANOVA analyzing using SPSS 18.0 statistical software.P value of ＜0.05 was considered to indicate statistical significance.Results 1.Cytotoxicity assay showed that cytotoxicity was not found in the relevant range of ART concentrations under 62.5 μmol/L.TC0 and TC50 value of ART were 62.5 μmol/L and 171.7 μmol/L.2.In concentration of 5 μmol/L,15 μmol/L and 30 μmol/L,ART and GCV could obviously inhibit growth of HCMV AD169 strains.There was no significant difference between them.The value of GCV IC50 was 3.49μmol/L,and the value of ART IC50 was 2.17 μmol/L.Treatment index (TI) of ART was 28.8,and GCV was 716.3.ART could still obviously inhibit growth of HCMV resistant strains,but GCV couldn't.Differences between them were statistically significant.The value of GCV IC50 to HCMV resistant strains was 44.4 μmol/L,and the value of ART IC50 was 2.5 μmol/L.3.Fractionation dosage method (2 times a day) of ART could improve the inhibition rate of virus significantly compared to that used once a day and single dose method.Difference was statistically significant(P ＜ 0.01).GCV delivered as the same method had little different changes in virus suppression ratio(P ＞ 0.05).Conclusions 1.Cytotoxicity was not found in the relevant range of ART concentrations under 62.5 μmol/L.2.ART could obviously inhibit growth of HCMV resistant strains and standard laboratory strains.3.Fractionation dosage method (2 times a day) of ART could improve the inhibition rate of virus significantly compared to that used once a day and single dose method.4.Because the action mode of ART is different from other anti-HCMV drugs,and ART has a high biological activity and fewer side effects,it is expected to become a kind of new antiviral drugs for HCMV infections.

This study was aimed to investigate the effect of baicalin on the proportion of Th22 cells and the concentration of IL-22 bothin vivo andin vitro, in order to explore the immune mechanism of baicalin on inflammatory bowel disease mice model. The 3.5% dextran sodium sulfate (DSS) was used on C57BL/6 mice for the establishment of colitis mice model. Mice were randomly divided into the blank control group, model group, and baicalin group. Flow cytometry and ELISA were used in the detection of the proportion of Th22 cells and concentration of IL-22 in peripheral blood serum, respectively. The spleen lymphocytes of mice were isolated and cultured by baicalin medium (0, 10, 20, 40μM) for 48 h. Flow cytometry was used in the detection of the proportion of Th22 cells. The results showed that baicalin reduced the proportion of Th22 cells and the expression of IL-22 bothin vivo andin vitro experiments. It was concluded that baicalin can inhibit Th22 cell differentiation and expression of IL-22in vitro and DSS-induced colitis mice. It indicated that baicalin had a good treatment potential in Th22 cell-mediated inflammatory diseases.

AIM: To investigate the dynamic variety of frequency and function of FoxP3~+ regulatory T cells in patients with acute hepatitis B (AHB). METHODS: Peripheral blood mononuclear cells (PBMCs) from 15 AHB patients at acute phase (week 1 of illness), convalescent phase (primary occurrence of both ALT level normalization and HBsAg negative conversion), resolved phase (at least 8 weeks after both ALT normalization and HBsAg seroconversion, and 15 health subjects were analyzed for FoxP3 (Forkhead/winged helix transcription factor) mRNA expression in MACS magnetic beads-purified CD4~+ T cells by real-time RT-PCR assay. The effects of Treg cells on the proliferation of CD4~+CD25~- T cells were examined by a ~3[H]-thymidine incorporation assay. RESULTS: AHB patients presented a significantly higher FoxP3 mRNA expression at convalescent phase than acute phase (t=-6.04, P<0.01) and resolved phase (t=4.45, P<0.01), and healthy controls (t=3.44, P<0.01). We also observed that the suppression efficiency of Treg cells on proliferation of CD4~+CD25~- T cells was lower at acute phase than convalescent phase (t=-5.30, P<0.01) and resolved phase (t=-3.20, P<0.05), but there was no significant difference between healthy controls and any phase of AHB. CONCLUSION: AHB patients presented lower circulating Treg frequency and suppression function at acute phase, and both of them are increased at convalescent phase, and then return to normal level along with disease resolved. This follow-up study furthers our understanding of Treg' s role in immunopathogenesis of hepatitis B.

Objective To investigate the changes in the glutamate transporter EEAC1 mRNA expression and the injured nerve cell apoptosis after rat cerebral ischemia/reperfusion injury is treated with mild hypothermia. Methods The middle cerebral arteries(MCA)of Sprague-Dawley rats were occluded for 30 minutes,and reper- fused for 90 minutes.Using DIG-labeled CRNA probe and TUNEL,the positive rate of glutamate transporter EE- AC1 mRNA expression and apoptotic cell rate were determined in the sham-operated group,the control group and the mild hypothermia group,respectively.Results The positive rate of EEAC1 mRNA expression and apoptotic cell rate were significantly lower in sham-operated group than those in the control group(P

ObjectiveTo investigate the genetic polymorphisms of 21 short tandem repeat(STR)loci (D3S1358, D13S317, D7S820, D16S539, Penta E, D2S441, TPOX, TH01, D2S1338, CSF1PO, Penta D, D10S1248, D19S433, vWA, D21S11, D18S51, D6S1043, D8S1179, D5S818, D12S391 and FGA). Methods A total of 560 blood samples were collected from unrelated healthy individuals of Han population in Hunan Province. Chelex-100 extraction method was applied to the extraction of genomic DNA, and an AGCU EX22 Kit and 9700 STR amplification was used in amplification reactions. The products were separated and analyzed on 310 Genetic Analyzer.ResultsA total of 248 alleles were observed, the al-lelic frequencies ranging from 0.001 to 0.518. Observation of genotype distributions for each locus showed no deviations from Hardy-Weinberg equilibrium exceptPentaE(P=0.023). The combined pow-er of discrimination, combined power of exclusion, and combined matching probability of the 21 STR loci were approximately 0.999 999 999 999 999 999 999 999 8, 0.999 999 998, and 1.36×10-25, respectively. ConclusionThe 21 STR loci show high polymorphisms in the Han population, which can provide valu-able data and a theoretical basis for forensic individual identification and paternity testing.

Objective To prepare high-titer monoclonal antibodies against STX2A1 subunit of enterohemorrhagic E.coli(EHEC) O157∶H7.Methods BALB/c mice were immunized with GST-STX2A1 fusion protein and the spleen cells of BALB/c mice which were not immunized were used as feeder cells.Hybridoma technique,natural STX2A protein and ELISA test were used to prepare and screen the hybridoma cell lines of monoclonal antibodies against STX2A1.The ascites developed by injecting the hybridoma cells into abdominal cavity of the BALB/c mice and was purified with Protein A-Sepharose.The subclasses and isotypes were identified by mouse monoclonal antibody isotyping kit.The antigenic epitopes that can be recognized by STX2-1A3,STX2-1E10 and STX2-3A7 were analyzed by the ELISA additivity test.Results Three hybridoma cell strains were obtained and named as STX2-1A3,STX2-1E10 and STX2-3A7,respectively,all of which produced monoclonal antibodies specifically against STX2A1.The isotypes of the monoclonal antibodies were IgG1?,IgG1?,and IgG3? and the affinity constant was 5.76 ?109,1.21 ?109 and 3.97 ?108,respectively.Conclusion We have successfully prepared three hybridoma cell strains which secrete high-titer and highly specific monoclonal antibodies against STX2A1.Our study provides a basis for researching the early diagnosis,prevention and cure of the disease induced by EHEC O157∶H7.

OBJECTIVE: To investigate and analyze the changes of postmortem human biochemical indexes. METHODS: Subclavian venous blood samples were collected from 81 cases of traffic fatalities. Thirteen blood biochemical indexes including liver function (ALT, AST, TBIL and DBIL), renal function (UA and Cr), cardiac function (CK, CK-MB and LDH), electrolytes (K+, Na+ and Cl-), and glucose (GLU) were tested by Roche cobas c311 automatic biochemical analyzer. The descriptive analysis was made by SPSS 17.0 statistical software. RESULTS: The values of ALT, AST, CK, CK-MB, LDH and K+ were higher than normal reference values with more fluctuations. The values of TBIL, DBIL, UA, Cr, Na+, Cl- and GLU were relatively stable with less fluctuations. CONCLUSION The postmortem human blood biochemical indexes of liver function, renal function, cardiac function, electrolytes and glucose could be affected by the factors, especially hemolysis and autolysis. The biochemical indexes, particularly enzymes, increased significantly with higher standard deviation.
Accidents, Traffic/mortality* ; Autopsy ; Blood Chemical Analysis/methods* ; Heart Function Tests ; Humans ; Kidney Function Tests ; Liver Function Tests ; Reference Values

0.05).Conclusions ATP-TCA could be used to individualize chemotherapy by selecting agents for particular patients of cervical cancer.The expression of GST-? and TS protein might be useful biomarkers to predict the resistance to DDP and 5-FU in patients with cervical cancer.