Objective To explore the effect of Melatonin(MT) on CD4+CD25+ regulatory T cell (CD4+CD25+Tr)and airway inflammation in asthmatic rat.Methods Thirty-two SD rats were randomly divided into 4 groups,8 rats in each group.Asthmatic group:rats were immunized on day 1 and 7 by intraperitoneal inject of mixture of ovalbumin(OVA) and aluminumhydroxide.From day 14,the animals were allenged with aerosolized OVA for 20 min per day for 7 consecutive days.MT group:OVA-sensitized rats were injected intraperitoneally with 0.1 mg/kg MT 30 min before each OVA challenge.Dexamethasone group:OVA-sensitized rats were injected intraperitoneally with 0.5 mg/kg Dexamethasone 30 min before each OVA challenge.Control group:OVA for inhalation and MT for intraperitoneal injection was replaced with saline.After the last challenge,peripheral blood was stained to count the percentage of eosinophil(EOS).Then the rats were lavaged and total leukocytes counts in bronchoalveolar lavage fluid(BALF) were performed after staining with Wright-Giemsa staining.The EOS counts around the airway was counted after the histological section of lung staining with hematoxylin and eosin staining.The serum level of immunoglobulin E(IgE) was detected by immunoenhancement.The change of CD4+CD25+Tr was assessed with flow cytometry.SPSS 10.0 software was applied to analyze data. Results In asthmatic rats,the CD4+CD25+ Tr/ CD4+T cells ratio had significant negative relationship with the EOS counts around the airway and the total leukocytes counts in BALF (r=-0.73 P0.05).There was a significant decrease in the percentage of the eosinophils in peripheral blood,the eosinophil counts around the airway,the total leukocytes counts in BALF and the serum level of IgE in MT group compared with asthmatic group (Pa

Objective To evaluate a new system,Vitek 2 Compact,for identification of bacteria and yeasts.Methods 185 clinical isolates of Peking Union Medical College Hospital,including 69 gram- positive strains,66 gram-negative strains and 50 yeasts,and 50 reference strains in our laboratory,including 25 gram-positive and gram-negative strains respectively,were studied.All the strains were identified by Vitek 2 Compact with GP,GN or YST identification cards.The API method was used as the reference method.Results Among the 93 gram-positive strains,85 strains(91.40%)were correctly identified, including 5 low discrimination identified strains,and 8 strains(8.60%)were correctly identified to the genus level,but misidentified to the species level.About 90% of gram-positive strains were identified within 7 h.Out of 91 gram-negative strains,90 strains(98.90%)were correctly identified,with 5 low discrimination identified strains,only 1 strain(1.1%)was correctly identified to the genus level,but misidentified to the species level.Above 90% of Enterobacteriaceae were identified within 5 h,and over 90% of nonfermenting bacteria were identified within 10 h.In the 50 strains of yeasts,46 strains(92%) were correctly identified,including 8 low discrimination identified strains,and 4 strains(8%)were correctly identified to the genus level,but misidentified to the species level.In all the yeasts,45 strains (90%)were identified in 18.25 h,and another 5 strains(10%)were identified in 18.50 h.Conclusions As Vitek 2 Compact system can give us reliable identification results of clinically relevant bacteria and yeasts,together with its significant reduction of handling time,it will definitely become a powerful tool in clinical microbiology laboratory.

Objective To compare BACTEC Plus aerobic and anaerobic bottles with BacT/ALERT FA aerobic bottles and FN anaerobic bottles in the ability of detecting simulated bacteremia specimens.Methods The 202 pairs of specimens were composed of 5ml sterile blood and defined loads of microorganisms.112 pairs of specimens in them also contained defined doses of antibiotics to simulate the patients undertaking antibiotic therapy.Time-to-detect(TTD)and positive percentages were evaluated in four groups,including aerobic bottles detecting aerobic bacteria,anaerobic bottles detecting anaerobic bacteria and facultative anaerobic bacteria,and aerobic bottles contained antibiotics detecting aerobic bacteria.Results The positive percentages of two kinds of aerobic bottles were both 100%.For the specimens with bacterial concentration of 10~2 and 10~3 CFU/ml,average TTDs of BACTEC Plus aerobic bottles[(13.69?3.74)h,(11.54?2.87)h]were faster than those of BacT/ALERT FA bottles [(16.76?5.62)h,(14.47?4.30)h;t=-5.674,-7.294,P

It is an important task to lab of clinical microbiology to surveille multi-drag or pan-drug resistant strains,such as penicillin-or macrolide-resistant Streptococcus pneumoniae,methicillin-resistant Staphylococcus aureus(MRSA),3rd generation cephalosporin-or quinolone-resistant Enterobacteriaceae, carbapenem-resistant Pseudomonas aeruginosa or Acinetobacter baumannii,and so on.We must understand characteristics of these resistant strains to guide doctors' empirical therapy of infective diseases.

Objective To evaluate a new system,Vitek 2 Compact,for antimicrobial susceptibility testing(AST)of gram-negative and gram-positive bacteria.Methods Eighty-nine clinical isolates of Peking Union Medical College Hospital,including 48 gram-negative strains and 41 gram-positive strains,and 66 reference strains kept in our laboratory,including 41 gram-negative strains and 25 gram-positive strains, were studied.The antimicrobial susceptibility of these strains were tested by Vitek 2 Compact with AST- GN09(for gram-negative bacteria),AST-P536(for Staphylococci),AST-P534(for Enterococci and S.agalactiae),and AST-P533(for S.pneumoniae)susceptibility cards.The Etest was used as the reference method for comparision.Thirty-two ESBL-producing strains assessed with the confirmatory tests for ESBLs of CLSI(16 strains of them had been confirmed by PCR amplified and sequencing)were detected for ESBLs by Vitek 2 Compact.Results According to the breakpoints of Clinical and Laboratory Standards Institute (CLSI),for the 1 626 microorganism-antibiotic combinations,Vitek 2 Compact gave 90.83% strains with category agreement(CA),4.91% strains with very major errors(VME),2.09% strains with major errors (ME),6.40% minor errors(MIE).The AST for more than 90% of Enterobacteriaceae,nonfermenting bacteria,micrococci and streptococci were completed within 11h,13h,11h and 12h,respectively.The ESBLs tests for thirty-two strains by V-itek 2 Compact are all positive.Conclusions Vitek 2 Compact system can give rapid,reliable and reproducible result with high sensitivity and specificity in assessment of antimicrobial susceptibility testing for clinically relevant gram-negative and gram-positive bacteria,and would become a powerful tool in clinical microbiology laboratory.

Objective To compare the antimierobial resistance of the gram-negative isolates from blood stream between Beijing and Hong Kong patients.Methods Non duplicate blood stream isolates were collected between 1998 and 2003 from Peking U- nion Medical College Hospital(n=530)and Hong Kong Prince of Wales Hospital(n=2913).Disk diffusion method and broth microdilution(MB)method were used to determine the antimierobial susceptibility of these strains to 13 antimicrobial a- gents including imipenem,ceftazidime,cefepime,etc.Results The resistance rates of E.coli to amikaein,ceftazidime and cefepime were 0.6%-5.0% and 7.5%-23.5%,and to gentamicin,cefuroxime,ciprofloxacin and cefuroxime were 12.6%- 28.4% and 29.4%-68.4% in Hong Kong isolates(n=1471)and Beijing isolates(n=213),respectively.The resistance rates of Klebsiella spp.to piperacillin-tazobactam and ceftazidime were 3.3%-11.3% and 4.3%-17.7%,and to amikacin, gentamicin,cefotaxime and ceftriaxone were 1.5%-10.1% and 5.8%-27.1% in Hong Kong isolates(n=586)and Beijing i- solates(n=70),respectively.The strains above and the ECS group of bacteria in Hong Kong(consisting of Enterobacter spp.,Citrobacter spp.,and Serratia spp.,n=333)were all susceptible to imipenem and meropenem,but 4.6%(n=65)of the ECS group bacteria in Beijing were resistant.About 8.8% and 14% of ECS group isolates were resistant to cefepime.Such isolates resistant to the other?-lactams ranged from 28.4% to 96%.In Hong Kong,Salmonella typhi remained susceptible to most of the antimierobial agents tested,but in Beijing,the resistance rates to ampicillin and piperacillin were from 9.1% to 18.2%.In Hong Kong,The resistance rates of Acinetobacter spp.and P.aeruginosa to imipenem and meropenem were lower than 10%,but in Beijing their resistance rates to imipenem were 13.6%-15.3% and 8.8%-30.8%,respectively.Conclusions The resistance rates of gram-negative bacteria isolated from blood stream in Beijing to most antimicrobial agents were higher than the corresponding rates in Hong Kong.This result will be useful for empirical therapy of local bacteremia.

Twenty Newcastle disease virus(NDV)strains were isolated from diseased chicken and geese in field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi,and the antigenic analysis of the all NDV isolates had been done based on the reaction spectrum with a panel of monoclonal antibodies to the HN glycoprotein.The entire ORFs encoding HN protein of these NDV isolates were amplified by RT-PCR successfully,cloned and sequenced.The resultant sequences of HN genes of 13 isolates of chicken origin and 7 isolates of goose origin were gained and analyzed.The results of reaction spectrum showed that there were some distinct differences in the antigenic epitopes among the 20 NDV isolates.And the sequences revealed that the coding regions of the HN genes of these isolates all consisted of 1716 nt characteristic of virulent strains of NDV,coding for 571 amino acids.Neucleotides sequence homology were found to be from 94.8%to 100%among 18 NDV isolates of genotypeⅦ,and the neucleotides sequence homology between all the isolates and the other genotypeⅦstrains of recent years in China ranged from 92.1%to 99.6%.The deduced amino acid sequences and the receptor-binding regions of HN proteins between the NDV isolates of chicken origin and of goose origin were compared and analyzed.The results showed that some unique amino acid substitutions were found in the genome of the NDV isolates,and the close genetic similarity provided evidence for epidemiological linkage between the NDV isolates of chicken origin and of goose origin in the same period.

OBJECTIVETo evaluate the clinical efficacy and safety of Huayu Tongbi Recipe (HTR) combined methotrexate (MTX) in treating refractory rheumatoid arthritis (RRA).

METHODSTotally 167 RRA patients were assigned to the treatment group (73 cases) and the control group (94 cases) according to different therapeutic methods. Patients in the treatment group were treated with HTR combined MTX, while those in the control group were treated with leflunomide (LEF) combined MTX. Clinical signs and symptoms, RF, CRP, ESR, disease activity score 28 (DAS28), and safety indicators were compared between the two groups before treatment, at week 12 and 24 after treatment. The efficacy and safety indices were also evaluated.

RESULTSAt week 12 after treatment the total effective rate was 82.2% (60/73 cases) in the treatment group and 79.8% (75/94 cases) in the control group, showing no statistical difference between the two groups (chi2 = 0.15, P > 0.05). At week 24 after treatment the total effective rate was 78.1% (57/73 cases) in the treatment group and 755% (71/94 cases) in the control group, showing no statistical difference between the two groups (chi2 = 0.15, P > 0.05). There was statistical difference in the total effective rate between week 24 and week 12 in the control group (chi2 = 0.49, P < 0.05). Clinical signs and symptoms, RF, CRP, ESR, and DAS28 were significantly improved in the two groups after 12- and 24-week treatment (P < 0.01). There was no statistical difference in the improvement at week 12 after treatment between the two groups (P > 0.05). There was statistical difference in time of morning stiffness, tender joint numbers, swollen joint numbers, patient global assessment, RF, CRP, and DAS28 at week 24 after treatment between the two groups (P < 0.05). Besides, adverse reactions occurred less in the treatment group than in the control group (P < 0.01).

CONCLUSIONThe efficacy of HTR combined MTX was equivalent to that of LEF (10 mg per day) combined MTX, but with more stable therapeutic effects and less adverse reactions.

Antirheumatic Agents ; pharmacology ; therapeutic use ; Arthralgia ; Arthritis, Rheumatoid ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Isoxazoles ; Methotrexate ; pharmacology ; therapeutic use ; Phytotherapy ; Treatment Outcome

OBJECTIVETo investigate the influence of pyrrolidine dithiocarbamate (PDTC) on the expression of transforming growth factor beta(1) (TGF-beta(1)), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor-1 of metalloproteinase (TIMP-1) in rats with pulmonary damage induced by paraquat (PQ).

METHODSFifty-four healthy male SD rats were randomly assigned into the control group (normal saline), the PQ-treatment groups (4 groups) and the PDTC treatment groups (4 groups). Except the rats in the control group, the rats in the PQ group were gavaged only with 40 mg/kg PQ, and PDTC group with 40 mg/kg PQ plus immediate injection 120 mg/kg PDTC (i.p). On the 3rd, the 7th, the 14th and 28th day after treatments, one group rats of each treatments were sacrificed and lung and blood samples were collected. The level of TGF-beta(1) protein in the plasma, the mRNA expression of TGF-beta(1), MMP-2 and TIMP-1 were evaluated using RT-PCR and real-time quantitative PCR, while pathological changes of lung were examined under optical microscope and electrical microscope.

RESULTSThe TGF-beta(1) protein, TGF-beta(1) and MMP-2 mRNA expression were increased significantly in the earlier stage and then decreased after PQ administration (P < 0.05 or P < 0.01), while the mRNA level of TIMP-1 was augmented continuously (P < 0.01) throughout the study compared to the control group. In comparison with the PQ group, in the PDTC treatment group, the TGF-beta(1) mRNA expression on the 3rd and the 14th day, 0.54 +/- 0.08 and 0.72 +/- 0.04 respectively, the MMP-2 mRNA expression on the 7th and 14th day, 1.62 +/- 0.50 and 1.97 +/- 0.34 respective-ly, and the TIMP-1 mRNA on the 7th and 21st day, 1.79 +/- 0.21 and 2.00 +/- 0.34 respectively, were significantly decreased (P < 0.05 or P < 0.01).

CONCLUSIONPDTC could attenuate paraquat-induced up-regulation of TGF-beta(1) and its mRNA expression, MMP-2 and TIMP-1 mRNA levels, which indicates that PDTC may exert its protective effects on paraquat-induced pulmonary damage by alleviating the earlier inflammation damage and adjust-ing the balance between MMPs and TIMPs. However, further studies are still warranted to investigate and clarify the underlying mechanisms involved in this complicated process.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Paraquat ; poisoning ; Pyrrolidines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo explore the therapeutic mechanisms of interferon-beta (IFN-beta) and intravenous immunoglobulin (IVIG) for experimental peripheral neuropathy induced by Campilobacter jejuni (Cj) lipopolysaccharide (LPS).

METHODForty healthy Wistar rats weighing 205 - 230 g were divided into IFN-beta, IVIG, IFN-beta plus IVIG and control groups. After the immune neuropathy was induced in the rats by Cj LPS, IFN-beta (1.3 microg/kg) was given by subcutaneous injection to the rats every other day for 6 weeks; IVIG [400 mg/(kg x d)] was given to the rats for five days, every other week for two times and IFN-beta [1.3 microg/(kg x d)] and IVIG [400 mg/(kg x d)] were given to the rats on the same days. Meanwhile, the control group was given PBS. The sera were collected in the 2nd, 4th and 6th week after therapy, the titers of anti-GM(1) IgG, MMP-9 and TNF-alpha in sera of immunized rats were measured by ELISA; histological study of sciatic nerve was performed and IgG on sciatic nerve was detected by immunohistochemistry in the 6th week.

RESULTS(1) There were no significant differences in titers of anti-GM(1) IgG, MMP-9 and TNF-alpha among the 3 therapeutic groups and control group after therapy for 2 weeks (P > 0.05). (2) The titers of anti- GM(1) IgG, MMP-9 or TNF-alpha in the control group were much higher than those of the IFN-beta group, the IVIG group or the IFN-beta and IVIG group after therapy for 4 weeks (P > 0.01) and there were no significant differences in titers of antibody among the 3 therapeutic groups (P > 0.05); the titers of MMP-9 or TNF-alpha in the IFN-beta and IVIG group were lower than those of the IFN-beta group or the IVIG group (P < 0.05). (3) The titers of anti-GM(1) IgG, MMP-9 or TNF-alpha in the control group were much higher than those of the IFN-beta group, the IVIG group or the IFN-beta with IVIG group after therapy for 6 weeks (P > 0.01); the IFN-beta with IVIG group had much lower levels of all indexes than the IFN-beta group or the IVIG group (P < 0.01).

CONCLUSIONIFN-beta and IVIG showed therapeutic effects on immune peripheral neuropathy through inhibiting the humoral and cellular immunity simultaneously in the peripheral neuropathy induced by CJ LPS, treatment with combined IFN-beta and IVIG was more effective.

Animals ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulins, Intravenous ; therapeutic use ; Immunotherapy ; Interferon Type I ; therapeutic use ; Interferon-beta ; immunology ; therapeutic use ; Lipopolysaccharides ; pharmacology ; Peripheral Nervous System Diseases ; therapy ; Rats ; Rats, Wistar ; Recombinant Proteins ; Sciatic Nerve ; drug effects ; Tumor Necrosis Factor-alpha ; immunology