OBJECTIVE To understand the distribution and epidemiology of ESBLs in ceftazidime or cefotaxime-(resistant) clinical Enterobacter cloacae isolates.METHODS Twenty seven ceftazidime or cefotaxime-resistant(nonrepetitive) E.cloacae were collected from 27 patients hospitalized at the Huashan Hospital,Shanghai.PCR and(sequencing) were performed to understand the distribution of ESBLs in E.cloacae;rep-PCR was(performed) to(understand) the epidemiology of ESBLs in E.cloacae.RESULTS CTX-M-3 like(ESBLs) were the most prevalent in our study(48%);this was the first report of VEB-1-like ESBLs in the member of(Enterobacteriaceae) in China,and the first report of the ESBLs VEB-1-like and CTX-M-3-like in an isolate simultaneously;the majority of(ESBLs) producers exhibited the same rep-PCR pattern,but harbored different ESBLs gene.(CONCLUSIONS) In our study,ESBLs have become prevalent in clinical E.cloacae isolates,and become an important factor of E.cloacae isolates resistant to extended-spectrum beta(-lactams).

This study was purposed to investigate the expression of annexin II in patients with hematologic malignancies and its role in genesis and development of hematologic malignancies. The expression levels of annexin II in bone marrow cells from untreated 81 patients with acute leukemia, 6 patients with MM and 20 patients with iron deficiency anemia were detected by real-time PCR assay. The results showed that the expression of annexin II mRNA significantly increased in M(3) patients as compared with others, the expression of annexin II gene in groups M(5), MM, M(4) were higher than that of other groups except M(3) group, and there were no significant difference in expression of annexin II gene between M(1) + M(2) groups and controls. It is concluded that the expression of annexin II gene significantly increased in patients M(5), M(4), MM, who showed higher ratio of infiltration than other patients. It is inferred that the annexin II participates in invasion and infiltration of hematologic malignancies probably through enhancing the degradation of extracellular matrix by cells of hematologic malignancies.
Adolescent ; Adult ; Annexin A2 ; genetics ; Bone Marrow Cells ; metabolism ; Female ; Hematologic Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo prepare anti-von Willebrand factor A3 (vWF-A3) domain monoclonal antibodies(mAbs) which block vWF-A3 binding to collagen, and characterize their biochemical properties and functions.

METHODSBALB/c mice were immunized with purified recombinant vWF-A3 protein (rvWF-A3). Murine anti-human vWF-A3 mAbs were developed by standard hybridoma technology and identified with ELISA. The recognition of the mAbs with rvWF -A3 and reduced human vWF was identified by Western-blot. The effect of mAbs on binding of purified human vWF to human placenta or calf skin collagen III was studied with collagen binding inhibition test.

RESULTSA group of 30 murine anti-human vWF-A3 mAbs was obtained, from which 2 clones were identified as inhibitory ones and designated as SZ-123 and SZ-125. SZ-123 and SZ-125 could react specifically with human vWF and rvWF-A3 respectively, while neither of them reacted with rvWF-A1 and rvWF-A2. Western-blot showed that SZ-123 and SZ-125 could recognize a 27 x 10(3) band of rvWF-A3 and 2 reduced human vWF bands at 250 x 10(3) and 170 x 10(3). SZ-123 and SZ-125 not only inhibited the binding of purified human vWF (1.5 and 3.0 microg/ml) to human type III collagen and to calf skin collagen III in a dose dependent manner, but also inhibited the binding of plasma vWF from human, rhesus monkeys or Beagle dogs to the two collagens.

CONCLUSIONSZ-123 and SZ-125 are neutralizing mAbs against vWF-A3 domain and may have therapeutic potential as an antithrombotic agent.

Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Collagen ; immunology ; Mice ; Mice, Inbred BALB C ; von Willebrand Factor ; immunology

OBJECTIVETo explore the operative methods and effects of treatment of old acetabular fractures.

METHODSFrom October 2001 to October 2007, 26 patients with old acetabular fractures were treated with operation including 21 males and 5 females with an average age of 34 years ranging from 18 to 65 years. On the basis of the three-dimensional computed tomography, all cases were diagnosed and classified according to Letourne-Judet classification, 9 cases were posterior wall fracture, 3 cases were lateropulsition fracture, 7 cases were lateropulsition and posterior wall fracture, 2 cases were posterior column and posterior wall fracture, 2 were T-shape fracture, 3 were dual column fracture. These patients were treated through the anterior,posterior, combined anterior-posterior approaches. The time from injured to operation was 33 to 141 days (averaged 36.4 days). All the fractures were fixed with screws and AO reconstruction plates.

RESULTSAll patients were followed up for 6 to 96 months, with an average time of 32.4 months. Evaluated according to Matta criteria, the results of scores was (5.04 +/- 1.04) on pain, (5.23 +/- 0.76) on range of motion, (4.92 +/- 1.16) on walking,and tatal (5.06 +/- 0.99) on average; The functional results of hip joints were excellent in 6 cases, good in 10 cases, fair in 6 cases, and poor in 4 cases. Sciatic nerve injury was found in 2 patients,lateral femoral cutaneus nerve injury in 3 patients, necrosis of femoral head in 1 patient,infection in 1 patient, and ectopic bone formation in 6 patients.

CONCLUSIONGood clinical results can be obtained by careful selection of operative indications of old acetabular fractures in combination with proper operative approach and correct reduction and fixation.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Time Factors ; Young Adult

OBJECTIVETo develop a monoclonal antibody (mAb) directed to FVIII C2 domain and investigate its effect on FVIII activity.

METHODSFVIII C2 protein was expressed in E. coli and purified. A murine antihuman FVIII C2 domain mAb SZ-132 was developed by standard hybridoma technology and characterized. In coagulation assays, different concentrations of SZ-132 were incubated with freshly collected pooled human plasma and the residual activity of FVIII and activated partial thromboplastin time (APTT) were determined. The effects of SZ-132 on rhFVIII binding to purified human vWF, phosphatidylserine (PS) and platelets were assessed by enzyme linked immunosorbent assays (ELISA).

RESULTSSZ-132 could inhibit FVIII procoagulant activity in a dose-dependent manner within the concentrations of 0-25 microg/ml and the FVIII activity was completely inhibited on above 25 microg/ml. It could also prevent rhFVIII from binding to vWF, PS and platelets.

CONCLUSIONSSZ-132 is a neutralizing mAb against FVIII C2 domain and can inhibit FVIII procoagulant activity by preventing FVIII from binding to vWF and PS.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Neutralizing ; biosynthesis ; immunology ; Factor VIII ; immunology ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C

OBJECTIVETo construct prokaryotic expression systems of ltB/ctB-ompL1/1 fusion genes and to determine the L.interrogans carrying status in leptospirosis patients with the expression products.

METHODSThe fusion genes ltB-ompL1/1 and ctB-ompL1/1 were constructed using linking primer PCR method. SDS-PAGE was used to examine expression of the target recombinant proteins rLTB-rOmpL1/1 and rCTB-rOmpL1/1. Western blot and GM1-ELISA were used to measure the immunogenic and GM(1)-binding activities of rLTB-rOmpL1/1 and rCTB-rOmpL1/1, respectively. PCR and MAT were performed to detect the expression of ompL1 gene in 97 wild L.interrogans strains. Antibodies against ompL1 gene products in serum samples of 228 leptospirosis patients were detected with ELISA method.

RESULTSThe homogeneity of nucleotide and putative amino acid sequence of ltB-jompL1/1 and ctB-ompL1/1 fusion genes were 99.7 % - 99.9 % and 99.5 % - 100 %, in comparison with the reported corresponding sequences. Expression outputs of both rLTB-rOmpL1/1 and rCTB-rOmpL1/1, mainly present in inclusion body, accounted for 10% of the total bacterial protein. Both rLTB-rOmpL1/1 and rCTB-rOmpL1/1 could combine to rabbit anti-rOmpL1/1 serum and bovine GM(1). 89.7 % of L.interrogans wild strains had ompL1 gene. 87.6% of the wild L.interrogans strains presented positive results for MAT (titers :1:4-1:256) with the rabbit anti-rOmpL1/1 or anti-rOmpL1/2 sera. 86.8% and 88.6% of the patients' serum samples were positive for rOmpL1/1 and rOmpL1/2 antibodies, respectively.

CONCLUSIONThe fusion proteins, rLTB-rOmpL1/1 and rCTB-rOmpL1/1, showed high immunogenic and GM(1)-binding activities. ompL1 gene is extensively distributed and frequently expressed in different serogroups of L.interrogans and its products expressed by different genotypes exhibit extensive cross-antigenicity.

Bacterial Outer Membrane Proteins ; genetics ; immunology ; Bacterial Toxins ; genetics ; Bacterial Vaccines ; genetics ; Cloning, Molecular ; Enterotoxins ; genetics ; Escherichia coli Proteins ; genetics ; Genes, Bacterial ; genetics ; Humans ; Leptospira interrogans ; genetics ; immunology ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; Vaccines, Synthetic ; genetics

The paper is to report the establishment of an HPLC specific chromatogram of Glycyrrhiza in Sini decoctions and the influence of combination on the specific chromatogram. The RP-HPLC method was used with a Phenomenex Gemini C18 column (250 mm x 4.6 mm ID, 5 microm), and acetonitrile-0.05% trifluoroacetic acid (gradient elution) as mobile phase. Flow rate was 0.8 mL x min(-1) and the detection wavelength was set at 232 nm. The temperature of column was 30 degrees C. The method is stable and reliable with a good reproducibility, it can be used to determine the specific chromatogram of Glycyrrhiza in Sini Decoctions. Twenty peaks were selected as specific peaks in Sini Decoction with liquiritin peak as the reference peak. Six of them were from Glycyrrhiza and the other 6 peaks were from both Glycyrrhiza and Ganjiangfuzi Decoction. The areas of specific peaks of Sini Decoctions were smaller than those in the chromatogram of Glycyrrhiza. The specific chromatogram of Glycyrrhiza in Sini Decoctions is markedly influenced by Radix Aconiti Carmichaeli and Rhizoma Zingiberis. The areas of the specific peaks in Sini Decoctions were reduced obviously. The method is stable and reliable with a good reproducibility, it can be used to determine the specific chromatogram of Glycyrrhiza in Sini Decoctions.
Aconitum ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; chemistry ; Ginger ; chemistry ; Glucosides ; chemistry ; Glycyrrhiza ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results

OBJECTIVETo observe the effect of a new synthetic tripeptide [Arg(NO(3))- Lys(OCH(3))- Arg(NO(3))] on L-arginine/NO pathway in the macrophage cell strain RAW246.7.

METHODSThe cultured macrophages exposed to lipopolysaccharide (LPS, 1 microg/L) treatment were randomly divided into 3 groups (n=6) and treated with distilled water, 1x10(-4) mol/L tripeptide and 1x10(-4) mol/L L-arginine, NG-monomethyl-L-arginine (L-NMMA) for 24 h, respectively. The macrophages were incubated for 24 h with LPS (1 microg/L) and in the presence of different concentrations of L-arginine (0 to 2 mmol/L), or in normal culture medium (containing 0.5 mmol/L L-arginine) for 24 h with LPS (1 microg/L) and in the presence of tripeptide of 0 to 10x10(-4) mol/L. The changes of [(3)H]-L-arginine transport and NO production from the macrophages were measured.

RESULTNO release from macrophages incubated in the LPS-containing culture medium was 50 folds, and [(3)H]-L-arginine uptake 2.7 folds that in cells in normal culture medium. Tripeptide (1x10(-4) mol/L) inhibited [(3)H]-L-arginine transport and NO production by 67% and 71% respectively. The effect of tripeptide was stronger than L-NMMA (P<0.05). Extracellular L-arginine caused a concentration-dependent increase in nitrite production, which reached the maximum at concentrations above 0.5 mmol/L Km for nitrite production of 0.162+/-0.015 mmol/L and Vmax of 91.2+/-2.3 micromol/(24h.10(6) cells). L-arginine transport and NO production were inhibited by tripeptide, but their dose-dependent pattern of changes was different with EC50 of 0.21x10(-4) mol/L and 1.27x10(-4) mol/L, respectively.

CONCLUSIONSActivation of macrophages with LPS induces nitrite accumulation in the culture medium, which is dependent on the presence of extracellular L-arginine. The tripeptide induces dysfunction of L-arginine/NO pathway in macrophages, potently inhibits L-arginine transport and competitively combine the active sites of nitric oxide synthase.

Arginine ; metabolism ; Biological Transport ; drug effects ; Cells, Cultured ; Humans ; Lipopolysaccharides ; Macrophages ; cytology ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; antagonists & inhibitors ; Oligopeptides ; pharmacology

Objective To investigate the anesthesia residents' proficiency in the epidural puncture and training needs using questionnaire survey in China.Methods A questionnaire designed by ourselves was sent to anesthesia residents via the WeChat platform within 1 month.The data were recorded by the system automatically.Results A total of 795 anesthesia residents involved in the investigation,and the number of valid questionnaires was 753 (94.7％).There were 233 (30.9％) junior residents (0-2 yr of training),279 (37.1％) semi-senior residents (3-5 yr of training),and 241 (32.0％) senior residents (＞5yr of training).Compared with junior group,the difficulty score for epidural puncture was significantly decreased,and the confidence scores for performing normal middle lumbar,difficult lumbar,lateral lumbar and thoracic epidural puncture were increased in semi-senior and senior groups (P＜0.05).Compared with semi-senior group,the difficulty score for epidural puncture was significantly decreased,and the confidence scores for performing normal middle lumbar,difficult lumbar,lateral lumbar and thoracic epidural puncture were increased in senior group (P＜0.05).The self-evaluated difficulty of epidural puncture was lower as the number of prior epidural cases was more (r=-0.719,P＜0.01).There were 46.6％ of the residents who had received simulation-based training before performing epidural puncture on the patient,among which most residents considered the simulation-based training is effective in helping to familiarize with procedure (77.2％),familiarize with anatomy (70.4％),simulate the texture of different layers (47.9％),and enhance success rate of epidural puncture (56.7％).There were 75.0％ residents who considered visualization technology is helpful in enhancing the success rate and confidence of epidural puncture.Conclusion Currently,the proficiency of junior anesthesia residents in epidural puncture needs to be strengthened.The simulation-based training has not been widely applied in the epidural training,while residents think high of simulation-based training and are looking forward to visualization technique training.

OBJECTIVETo study the value of serum Cystatin C (Cyst C) in the evaluation of glomerular filtration function in children with viral encephalitis.

METHODSSerum levels of Cyst C, urea nitrogen (BUN) and creatinine (Cr) were measured in 92 children with viral encephalitis and in 50 healthy children as a control group. According to glomerular filtration rate (GFR), the encephalitis group was subdivided into normal renal function, renal insufficiency in the compensatory or decompensatory stage, and renal failure /end-stage groups.

RESULTSSerum levels of Cyst C, BUN and Cr in the encephalitis group increased and GFR decreased significantly compared with those in the control group (P<0.01). With the decline of renal function, GFR decreased and serum levels of Cyst C, BUN and Cr increased gradually. Serum levels of Cyst C and GFR were significantly different among the encephaitis subgroups (P<0.01). For serum levels of BUN and Cr, there were significant differences among the subgroups except between the normal renal function and the compensatory renal insufficiency groups. Serum Cyst C level was positively correlated with serum BUN and Cr levels, and negatively correlated with GFR.

CONCLUSIONSThe children with viral encephalitis have different degrees of renal impairments. Cyst C appears to be superior to BUN and Cr as a marker for the evaluation of glomerular filtration function. Measurement of serum Cyst C levels is very valuable in renal function monitoring in children with viral encephalitis.

Child ; Child, Preschool ; Cystatin C ; blood ; Encephalitis, Viral ; blood ; physiopathology ; Female ; Glomerular Filtration Rate ; Humans ; Infant ; Male ; Renal Insufficiency ; diagnosis