Objective To analyze the resistance genes in a muhidrug resistant Klebsiella pneumoniae (MDRKP) strain.Methods A MDRKP strain was isolated from bronchoalveolar lavage fluid in Pediatric Intensive Care Unit of Children's Hospital Affiliated to Soochow University in February 2012.Acquired resistance genes to beta-lactams,aminoglycosides,quinolones,ompK35 and ompK36 gene for outer membrane porin protein,and carbapenems targeting PBP2 gene were analyzed by PCR and verified by DNA sequencing.Results Acquired resistance genes TEM-1,SHV-1 to beta-lactam antimicrobial agents and aac(6′)-I b to aminoglycoside antimicrobial agents were positive in the strain of MDRKP.While 16S rRNA methylase,ompK35 and ompK36 genes for outer membrane porin protein were negative.Compared with susceptible strains,there were 9 synonymous mutations in PBP2 gene sequence of this MDRKP strain,but the amino acid sequences were the same.No mutation in quinolone resistance determining region (QRDR) was observed.Conclusion The multidrug resistance of the isolated Klebsiella pneumoniae strain may be related to 2 kinds of beta-lactam acquired resistance genes,1 kind of aminoglycoside acquired resistance gene,ompK35 and ompK36 genes defects and synonymous mutation in PBP2 gene.

Pancreatic cancer is a highly lethal disease in gastrointestinal malignant tumors.The mortality of pancreatic cancer closely parallels its incidence.Most patients with pancreatic cancer remain asymptomatic until the disease reaches an advanced stage.There is no program for screening patients at high risk of pancreatic cancer.Although CT,MRI,positron emission tomography,endoscopic ultrasonography,and endoscopic ultrasonography-guided fine-needle aspiration offer high diagnostic ability for pancreatic cancer,it cannot be found at the early stage easily.Surgical resection is regarded as the only potentially curative treatment and adjuvant chemotherapy is given after surgery.This article reviews epidemiology,risk factors,diagnosis and treatment for pancreatic cancer by summarizing relevant literature.

Objective: To investigate whether CD137 signaling can promote angiogenesis via regulating macrophage M1/M2 polarization. Methods: (1) The primary peritoneal macrophages in mice induced by 3% thiglycollate broth were divided into three groups: control group, CD137 signaling activated group and CD137 signaling inhibited group. Various specific markers of M1 and M2 macrophages were detected to observe the phenotype change of macrophages, and the macrophages protein expression of CD137, CD86 and CD206 was detected by flow cytometry (FCM). The protein and mRNA expression of induced nitric oxide synthase (iNOS), arginase Ⅰ(Arg-1) was determined by Western blot and RT-PCR, respectively. The secretion levels of IL-12 and IL-10 in culture supernatant of macrophages were detected by ELISA. (2) Macrophages were co-cultured with the endothelial cells (bEnd.3), and macrophages were implanted in the upper chamber, endothelial cells were implanted in stromal glue of the lower chamber. The experiment was divided into three groups: the control group, CD137 signaling activated group and peroxisome proliferator-activated receptor-γ (PPAR-γ) inhibited group, and tube formation ability of endothelial cells in each group was determined. Results: (1) The purity of primary peritoneal macrophages in mice was (97.93±1.31)%. The expression of CD137 on the surface of macrophages was (97.40±2.70)%. (2) Compared with control group, the mRNA and protein expression levels of Arg-1 were significantly increased and the mRNA and protein expression of iNOS were significantly decreased in CD137 signaling activated group (all P<0.05). Compared with CD137 signaling activated group, the mRNA and protein expression of Arg-1 were significantly lower and the mRNA and protein expression levels of iNOS were significantly higher in CD137 signaling inhibited group (all P<0.05). FCM results showed that the average fluorescence intensity of CD206 was higher, while the average fluorescence intensity of CD86 was lower in CD137 signaling activated group than in control group (P<0.05, P<0.01, respectively); the expression of CD206 was significantly lower, while the expression of CD86 was higher, in the CD137 signaling inhibited group than in CD137 signaling activated group (P<0.05, P<0.01, respectively). ELISA results showed that the secretion of IL-10 was higher, and the secretion level of IL-12 was significantly lower in CD137 signaling activated group than in control group (both P<0.01); the secretion of IL-10 was significantly lower and the secretion of IL-12 was significantly higher in CD137 signaling inhibited group than in CD137 signaling activated group (both P<0.05). (3) Values of the formation of tube length and branch number were both longer in CD137 signaling activated group than control group (P<0.05). The formation of the tube length and branch number were less in PPAR-γ inhibited group than in CD137 signaling activated group (P<0.05). Conclusion: CD137 signaling can promote angiogenesis by regulating macrophage M1/M2 polarization.
Animals ; Coculture Techniques ; Endothelial Cells ; Macrophages ; Mice ; Neovascularization, Pathologic ; Signal Transduction

This study was aimed to investigate the effect of PTD-mFoxp3 fusion protein on graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation. The 10-weeks-old C57BL/6 mice as recipients were randomly divided into three groups (A,B and C), 10 mice were in each group. The mice on day of transplantation as on day 0 received total body irradiation (TBI) 6.0 Gy, then the bone marrow cells (BMC) from BALB/c mice were injected through tail vein within 4-6 hours. At 2 days before transplantation and 0, 1, 3, 5, 7, 9 and 13 days after transplantation, mice in group A were injected with saline, mice in group B were injected with mFoxp3 protein and mice in group C were injected with PTD-mFoxp3 fusion protein. Symptoms of GVHD, survival time and histopathological changes were observed. The establishment of mixed chimerism was determined by flow cytometry in day 60, and IL-2 and IFN-γ expression profiles in the recipient peripheral blood were assessed by ELISA. The results showed that the mean survival time of recipients in group A,B and C was (32.95 ± 5.48) , (38.00 ± 5.45) and (55.30 ± 3.15) respectively. Graft rejection was observed in the liver and small intestine specimens of group A and group B. The serum levels of IL-2 and IFN-γ significantly decreased in the recipients of group C, as compared with the other groups. The flow cytometry analysis revealed that the survival recipient mice developed high chimerism levels, the percentages of donor cells in group A,B and C were (79.46 ± 1.80) %, (79.13 ± 2.23) % and (85.92 ± 2.82) % respectively. It is concluded that PTD-mFoxp3 fusion protein can reduce the incidence and mortality of GVHD after allogeneic bone marrow transplantation.
Animals ; Bone Marrow Transplantation ; adverse effects ; Female ; Forkhead Transcription Factors ; therapeutic use ; Graft vs Host Disease ; metabolism ; therapy ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Recombinant Fusion Proteins ; therapeutic use ; Transplantation, Homologous

OBJECTIVETo establish a RP-HPLC method to determine the pharmacokinetics of ferulaic acid of ferulaic acid, Rhizoma Chuanxiong extraction and Naodesheng capsule in rat and assess the effect of other components in medical material and in compound on the pharmacokinetics of ferulaic acid.

METHODThe rats were orally treated with referential ferulaic acid, Rhizoma Chuanxiong extraction and Naodesheng capsule repectively. Blood samples were collected by cutting rats tails. Plasma was separated by centrifugation at 10,000 r x min(-1) for 10 min, and two-times methanol in volume was added to deposit proteins. After centrifugation, the upper liquid was transferred to filter. The concentration of ferulaic acid in serum was determined by RP-HPLC. The stationary phase was C18, and methanol-0.5% acetic acid (30:70) was taken as the mobile phase, A UV detector was used at 320 nm. The pharmacokinetic parameters were calculated with 3p97 program.

RESULTA good linear relationship of ferulaic acid was obtained from 0.05-1 mg x L(-1), the lowest limits of determination were 13 microg x L(-1). The plasma concentration-time curves of ferulaic acid were fitted with two-compartment models properly. The pharmacokinetics parameterst AUC(0-t), Cmax, CL of ferulaic acid showed significant differences between referential group and the other groups.

CONCLUSIONThe method applied for determination of ferulaic acid content in blood was simple, accurate and feasible for the study of ferulaic acid pharmacokinetics in rats. The results indicated that the other ingredients of Rhizoma Chuanxiong had remarkable influence on the pharmacokinetics of ferulaic acid. However, compatibility promotes the ferulic's absorption, enhances the ferulic's biological exploitability.

Animals ; Capsules ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Pinellia ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVEPatient with myocardial bridging (MB) usually has a benign prognosis, but some MB patients might experience myocardial ischemia, infarction and sudden cardiac death, especially during active physical activities. The purpose of the study was to study the stress-induced blood flow changes of the mural coronary artery in MB patients determined by intracoronary Doppler.

METHODSIn 8 patients with MB, the basic average peak velocity (bAPV), hyperemic average peak velocity (hAPV) of blood flow, coronary flow reverse (CFR) proximal and distal to the mural coronary artery were measured before and during intravenously dobutamine (10 microg kg-1 min-1, then add 10 microg kg-1 min-1 at 3 min interval till 40 microg kg-1 min-1) by intracoronary Doppler.

RESULTSThe baseline mural coronary diameter reduction was (51.7+/-21.4)% and significantly increased to (90.0+/-12.7)% (P<0.01) during dobutamine infusion. bAPV on the segments proximal and distal to the mural coronary artery significantly increased from (19.83+/-5.84) cm/s and (20.75+/-4.91) cm/s to (31.52+/-10.93) cm/s and (30.46+/-9.01) cm/s (all P<0.05 vs. baseline) respectively post dobutamine infusion. CFR measured at proximal and distal to myocardial bridging also significantly decreased from (2.91+/-0.62) and (2.46+/-0.82) to (2.17+/-0.66) and (1.83+/-0.51) (all P<0.01).

CONCLUSIONStress can significantly increase the compression of intramural coronary artery and reduce CFR on coronary segments both proximal and distal to the MB. Thus, active exercise might induce myocardial ischemia in patients with myocardial bridging.

Blood Flow Velocity ; Cardiotonic Agents ; pharmacology ; Coronary Circulation ; drug effects ; Coronary Vessel Anomalies ; physiopathology ; Coronary Vessels ; drug effects ; Dobutamine ; pharmacology ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo assess the effect of beta blocker on blood flow velocity and reserve on the intramural coronary artery of patients with myocardial bridging.

METHODSIn 8 patients with myocardial bridge, intracoronary Doppler was performed before and after esmolol was given intravenously. The basic average peak velocity (bAPV), hyperaemic average peak velocity (hAPV) of blood flow, and coronary flow reserve (CFR) proximal and distal to the mural myocardial bridging was measured and compared.

RESULTSAfter esmolol injection, the mural coronary diameter systolic reduction decreased from (58.0 +/- 14.7)% to (26.0 +/- 9.8)% (P < 0.01); the bAPV proximal and distal to myocardial bridging separately decreased from (19.4 +/- 4.9) cm/s and (18.4 +/- 3.6) cm/s to (4.7 +/- 3.9) cm/s (P < 0.01) and (15.1 +/- 1.5) cm/s (P < 0.05). Under hyperemization, esmolol changed the hAPV of proximal and distal to myocardial bridging separately from (54.1 +/- 14.9) cm/s and (44.7 +/- 9.4) cm/s to (49.7 +/- 16.4) cm/s and (48.9 +/- 10.1) cm/s (all P > 0.05); thus, the value of CFR both proximal and distal to myocardial bridge increased separately from 2.8 +/- 0.3 and 2.5 +/- 0.5 to 3.4 +/- 0.5 and 3.2 +/- 0.6 (all P < 0.01).

CONCLUSIONEsmolol can decreased the compression of the intramural coronary artery and increased the CFR to normal level of it.

Adrenergic beta-Antagonists ; pharmacology ; therapeutic use ; Coronary Circulation ; drug effects ; physiology ; Coronary Vessels ; diagnostic imaging ; drug effects ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Myocardial Bridging ; drug therapy ; physiopathology ; ultrastructure ; Propanolamines ; pharmacology ; therapeutic use ; Ultrasonography, Interventional

The latest progress in research on constituents and pharmacological activities of sarcotestas of Ginkgo biloba has been studied. The main constituents in sarcotestas of G. biloba include flavones, ginkgolides, alkylphenols, polysaccharides and amino acids, etc. They show the following activities, such as bacteriostatic, bactericidal and pesticidal activities, antitumor and mutagenic, carcinogenic effects, antianaphylaxis and allergenic activity, effects on immunologic function, scavenging free radical, antisenile action, etc. The problems at present and the reseach direction for the future on sarcotestas of G. biloba have been put forward.
Animals ; Anti-Bacterial Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Flavonoids ; analysis ; isolation & purification ; pharmacology ; Free Radical Scavengers ; pharmacology ; Fruit ; chemistry ; Ginkgo biloba ; chemistry ; Ginkgolides ; analysis ; isolation & purification ; pharmacology ; Humans ; Plants, Medicinal ; chemistry ; Salicylates ; analysis ; isolation & purification ; pharmacology

To investigate the effect of monoamine oxidase inhibitor tranylcypromine (TCP) on the differentiation of human U251 glioma cells, we treated U251 cells with TCP and/or 100 nmol/L histone deacetylase inhibitor trychostatin A (TSA). The differentiation of U251 cells was observed with inverted microscopy. The cell proliferation and cell cycle distribution were determined by MTT assay and flow cytometry, respectively. Apoptosis was observed by Hoechst 33258 staining. The levels of differentiation-related genes were assessed by real-time PCR and Western blotting. TCP-induced differentiation was characterized by typical morphological changes, inhibition of cellular proliferation, accumulation of cells in the G1 phase of the cell cycle, decreased expression of the pluripotency transcription factors Oct4 and Sox2, and increased expression of glial fibrillary acid protein (GFAP). The combination of TCP and TSA treatment also triggered an over-expression of GFAP. These findings suggest that TCP may induce differentiation of U251 glioma cells, and the differentiation process may be promoted by histone deacetylase inhibitor TSA.
Brain Neoplasms ; pathology ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; drug effects ; Glioma ; pathology ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Hydroxamic Acids ; pharmacology ; Monoamine Oxidase Inhibitors ; pharmacology ; Tranylcypromine ; pharmacology

This study is to investigate the effects of puerarin on the proliferation and differentiation of umbilical cord mesenchymal stem cells (MSCs) into osteoblasts. Umbilical cord MSCs were cultured by tissue adherence and the third passage of cells was used in the experiment. The effect of puerarin on proliferation of umbilical cord MSCs was measured with MTT. The effects of puerarin on umbilical cord MSCs were evaluated by ALP immunohistochemisty and von kossa staining. The OD value decreased with the increase of puerarin concentration. On 7th day, ALP expression of puerarin group was higher than that of control group. On 14th day, ALP staining showed that the positive rate of puerarin group was higher than that of control group. Von kossa staining showed the quantity of calcium nodules was higher in puerarin group than that of control group. Puerarin can promote the umbilical cord MSCs to differentiate into osteoblasts and has an effect on the proliferation of umbilical cord MSCs.
Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Humans ; Isoflavones ; isolation & purification ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Osteogenesis ; drug effects ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Umbilical Cord ; cytology