OBJECTIVE To investigate the genotypes of plasmid-mediated AmpC beta-lactamases carried in Klebsiella pneumoniae,to provide reasonable use of antibacterial agents and to reduce the spread of these drug resistant genes.METHODS Two hundred and eighteen strains of K.pneumoniae were performed in Tongji Hospital in Wuhan.Plasmid-mediated AmpC beta-lactamases-producing strains were screened by improved three-dimensional method and identified by multiplex PCR.DNA sequencing method was used to confirm these drug resistant strains.The MIC of 15 kinds of antibacterial agents against the clinical isolates was detected by double agar dilution method.RESULTS Thirteen strains of K.pneumoniae were detected by improved three-dimensional method.The detection rates were 5.96%.There were seven of thirteen positive strains of K.pneumoniae through improved three-dimensional method harboring DHA-1 type plasmid-mediated AmpC beta-lactamase by multiplex PCR and confirmed by DNA sequencing.Plasmid-mediated AmpC beta-lactamase producers revealed a highly drug resistance to cephalosporins,monobactams,beta-lactam/beta-lactamase inhibitor combinations,aminoglycosides and fluoroquinolones.Only imipenem was susceptible to all of these detected strains.CONCLUSIONS DHA-1 plasmid-mediated AmpC beta-lactamase is detected in K.pneumoniae strains from Tongji Hospital.The detection rate is 3.2%.The pAmpC-producing strains reveal multi-drug resistance.Only imipenem is susceptible to them.

Pro-inflammatory macrophages play key regulatory role in the occurrence and development of rheumatoid arthritis (RA). In this study, we constructed a celastrol (Cel)-loaded polyamide-amine dendrimer (PAMAM) drug delivery system, which could target folate receptor and mitochondria. It could target inflammatory macrophages and realize chemo-photothermal synergistic therapy. Using PAMAM as the nano-carrier, folate receptor-targeting group folic acid (FA) and mitochondria-targeting group IR808 (also known as the photothermal agent) were conjugated with PAMAM through amide reaction, and then complexed with anti-inflammatory drug Cel to prepare the FA-PAMAM-IR808/Cel nanocomplex. In vitro characterization results showed that the drug loading efficiency of the nanocomplex was 50.90%, particle size was between 130 and 160 nm, average potential was between 1.0 and 3.5 mV, the drug release showed pH sensitivity, temperature reached to 42.5 ℃ after near-infrared (NIR) light irradiation for 10 min. In vitro cellular uptake experiments showed that the nanocomplex had obvious folate receptor-targeting and mitochondria-targeting ability. Following irradiation with NIR light, the cytotoxicity and cellular apoptosis enhanced. The secretion of pro-inflammatory factors tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 and nitric oxide (NO) decreased in a concentration-dependent manner. This study provided insights for the development of novel anti-RA nanomedicines.

OBJECTIVETo explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment.

METHODThe three kinds of explant of bud, leaf and calli were induced by colchicine treatment.

RESULTThe induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different.

CONCLUSIONInduction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

Chromosomes, Plant ; genetics ; Colchicine ; pharmacology ; Plant Leaves ; anatomy & histology ; genetics ; growth & development ; Plant Shoots ; anatomy & histology ; genetics ; growth & development ; Plants, Medicinal ; anatomy & histology ; genetics ; growth & development ; Polyploidy ; Salvia ; anatomy & histology ; genetics ; growth & development

The aim of the present study was to investigate the changes of lymphatic contraction after hemorrhagic shock in vitro and the underlying role of nitric oxide (NO). Rat thoracic duct segments were isolated at 0, 0.5, 1, 2 and 3 h after hemorrhagic shock. Using Pressure Myograph System, we determined contraction frequency (CF), end systolic diameter (ESD), end diastolic diameter (EDD) and passive diameter (PD) of isolated rat lymphatics under different transmural pressures (1, 3, 5, 7 and 9 cmH(2)O), then calculated contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) of lymphatics. The results showed that in several transmural pressures, lymphatic CF, TI and FPF were significantly higher in shock 0 h and shock 0.5 h groups than those in control group (sham operation group). With the development of shock, lymphatic CF, TI and FPF decreased significantly in shock 2 h and shock 3 h groups compared with those in control group. We further discovered the role of NO in the changes of lymphatic contraction after hemorrhagic shock. Under 3 cmH(2)O transmural pressure, the changes of lymphatic contraction in shock 0.5 h and shock 2 h groups were analyzed following the incubation with several NO-related drugs alone or in combination. And the results showed that NO donor L-Arg reduced CF, TI and FPF in shock 0.5 h group to the control levels, while soluble guanylate cyclase inhibitor ODQ suppressed the effect of L-Arg. Moreover, NOS inhibitor L-NAME elevated the CF, TI and FPF of 2 h shock lymphatics to the control levels, while phosphodiesterase inhibitor aminophylline (AP) suppressed the effect of L-NAME. These results suggest that the lymphatic contractile activity exhibits a biphasic change during hemorrhagic shock, increasing in early phase and declining in later stage. And NO plays a major regulating role in the biphasic change of lymphatic contraction in hemorrhagic shock rats via cGMP pathway.
Animals ; Cyclic GMP ; metabolism ; In Vitro Techniques ; Male ; Muscle Contraction ; Muscle, Smooth ; physiopathology ; Nitric Oxide ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology ; Thoracic Duct ; physiopathology ; Time Factors

OBJECTIVETo observe the effects of mesenteric lymph drainage on erythrocyte rheology and blood viscosity in hemorrhagic shock rats.

METHODSWistar rats were randomly divided into sham-shock group, shock group (establishing hemorrhagic shock model), drainage group (establishing hemorrhagic shock model plus drainaging shock mesenteric lymph from hypotension 1 h). At 3 h of hypotension or corresponding time, blood samples were harvested from the abdominal aorta for determining the erythrocytic parameters, erythrocyte electrophoresis, erythrocyte sedimentation rate (ESR) and blood viscosity, and the erythrocytes aggregation index and erythrocyte deformability index were calculated.

RESULTSCompared with the sham-shock group, the red cell contents, hematocrit (HCT), hemoglobin (Hb), mean corpuscular hemoglobin concentration (MCHC), erythrocyte electrophoretic rate and mobility, erythrocyte deformability index, whole blood viscosity, whole blood relative or reduced viscosity at low and high shear rates in shock group were observably lower, and mean corpuscular volume, electrophoretic time of erythrocyte, ESR, K value of equation and K value of emendation, erythrocytes aggregation index, plasma viscosity in shock group were increased markedly; the MCHC, erythrocyte electrophoretic rate and mobility, whole blood viscosity, whole blood relative viscosity at low and high shear rates in drainage group were reduced, and the red blood cell volume distribution width -SD (RDW-SD) was increased remarkably. At the same time, in drainage group, the HCT, RDW-SD, erythrocyte deformability index, whole blood viscosity and relative viscosity at low and high shear rates were higher, the ESR, K value of equation and K value of emendation, erythrocytes aggregation index, plasma viscosity were lower than that of shock group.

CONCLUSIONThe results indicate that the mesenteric lymph drainage could improve the erythrocyte rheological behavior, as a result, improve the hemorrheological properties in hemorrhagic shock rats.

Animals ; Blood Viscosity ; Drainage ; methods ; Erythrocyte Aggregation ; Erythrocyte Deformability ; Lymph ; Male ; Mesentery ; Rats ; Rats, Wistar ; Rheology ; Shock, Hemorrhagic ; blood ; therapy

Objective To establish a method for content determination of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province.Methods The contents of total flavones and polysaccharides in Hedysari Radix in different producing areas in Gansu Province were determined by ultraviolet spectroscopy with calycosin-7-glucoside and glucose as reference substance, and the wavelength was set at 260 nm and 484 nm.Results The contents were from 2.82 mg/g to 6.79 mg/g for total flavones and from 106.14 mg/g to 746.40 mg/g for total polysaccharides in Hedysari Radix in different producing areas in Gansu Province. The recoveries of total flavones and total polysaccharides were 97.96％ and 102.90％, respectively.Conclusion There was difference in contents of total flavones and polysaccharides of Hedysari Radix in different producing areas in Gansu Province, and the method of using ultraviolet spectroscopy is simple, reproducible, accurate and reliable, which can be preferably used as the method for content determination of total flavones and polysaccharides in Hedysari Radix.

[Objective]To investigate the etiology,clinical manifestation,treatment and prognosis of optic disc vas-culitis.[Method]Twenty-four eyes of 21 patients were enrolled in this retrospective study.Eye examinations,treatment, and effect were recorded.[Result]Six were male and 15 were female.The age was between 19 and 43 years old(average:28.7±1.6).85.7% of the patients referred to the clinic with mild to moderate decreased vision.Edema of the optic disc can be seen in both types while tortuous veins can also be found in type 2.Similar characteristics were noticed in OCT,FFA, and etc.With a follow-up of 4.52±0.98 months after treatment(prednisone:initial dose 1.0-1.2 mg/kg),the BCVA of the affected eyes improved significantly.[Conclusion]Optic disc vasculitis is affected by autoimmune disorder,infection,hy-perlipidemia,and etc.Edema of the optic disc with/without tortuous veins and retinal hemorrhage can be noticed.Similar diseases should be excluded in avoidance of misdiagnosis. Systemic examination and complete solution should be per-formed.Glucocorticoid helps to improve the visual function.The application of anti-VEGF is effective in secondary macu-lar edema.However,the long-term efficacy is awaiting being confirmed.

OBJECTIVETo investigate the curative effect of antiphlogistic agent series on treating chronic nonbacterial prostatitis (CNP).

METHODSOne hundred and sixty patients were randomized into 4 groups for an 8-week clinical observation: group A (oral antiphlogistic medicinal granules only), group B (oral antiphlogistic medicinal granules + retention enema), group C (oral antiphlogistic medicinal granules + rectal), and group D (antiphlogistic medicinal granules + rectally + hip bath). Single blind trials were employed.

RESULTSThe curative rates of the 4 groups were 37.5%, 57.5%, 52.5% and 82.5% respectively, while the total efficacy rates were 42.5%, 82.5%, 77.5% and 92.5% respectively. Compared with groups A, B and C, the curative rate of group D was significantly higher (P < 0.05). The difference in efficacy rates was slight between groups B and D (P < 0.05), but significant between groups A and C (P < 0.05).

CONCLUSIONSCombined treatment therapy can improve the effect of CNP treatment and clear away heat and toxic material. The antiphlogistic agent series, with the effect of motivating blood circulation and removing blood stasis, turned out to be an effective traditional Chinese medicine in treating CNP.

Adult ; Chronic Disease ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Prostatitis ; drug therapy

This study is to investigate the apoptotic induction effect of the combination of diosgenin and TNF-related apoptosis-inducing ligand (TRAIL) on non-small-cell lung cancer cell line A549 by using the Chou-Talalay method, and observe the mechanism of the combination. The apoptotic effect of diosgenin or TRAIL alone and their combination on A549 and normal cell line 293T proliferation was measured by MTT assay. Chou-Talalay method was used to evaluate the combination effect. Apoptosis was examined by Hoechst 33342 staining and flow cytometry assay. Western blotting detects the expression of apoptosis-associated proteins. Diosgenin or TRAIL alone can inhibit proliferation ofA549 in a concentration-dependent manner. According to the Chou-Talalay method, when f(a) = 0.1, CI > 1, when f(a) > 0.1, CI < 1. Combined with TRAIL, the IC50 of diosgenin decreases from 21.864 to 14.810 micromol x L(-1) (P < 0.05) on A549 cells. But for 293T cells, IC50 of diosgenin does not change significantly. As with Hoechst 33342 staining and flow cytometry assay, the apoptosis ratios also increased in the combination group. At protein expression level, combination-treated group displays increased Caspase-8, Caspase-9, Bid, Caspase-3 activation and PARP cleavage, significantly decreased Bcl-2 and increased Bax expression, and MAPK pathways were activated. The combination of diosgenin and TRAIL has synergistic effect on A549 cells.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diosgenin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; HEK293 Cells ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Signal Transduction ; TNF-Related Apoptosis-Inducing Ligand ; administration & dosage ; pharmacology

OBJECTIVETo observe the change of lymphatic reactivity to substance P (SP) during the process of hemorrhagic shock (HS) with a technique of lymphatic perfusion in vitro in this study.

METHODSMale Wistar rats were randomly divided into control group (surgical procedure only) and HS group (the rats in this group were further divided into five subgroups: shock 0 h, 0.5 h, 1 h, 2 h and 3 h groups after duplicating the HS model with method of bloodletting to mean arterial blood pressure was 40 mmHg through the femoral venous). Thoracic ducts were separated from HS rats at the corresponding time points in each group. A segment of thoracic duct was pressed and perfused in vitro at transmural pressure of 3 cm H2O, and then stimulated with gradient SP respectively. The end systolic diameter, end diastolic diameter, contraction frequency (CF) and passive diameter of isolated lymphatics were measured, while the contraction amplitude (CA), tonic index (TI) and fractional pump flow (FPF) were calculated, and the different values between pre- and post- administration of SP of CF, CA, TI and FPF were calculated and expressed as Delta CF, Delta TI, Delta CA and Delta FPF to further assess the reactivity of lymphatics.

RESULTSAfter SP incubation, the Delta CF, Delta TI, Delta CA and Delta FPF of 0 h- and 0.5 h shocked lymphatics were significantly increased when compared with that of control group on one or several concentrations. The Delta CF (at 3 x 10(-7) mol/L of SP) and Delta TI (1 x 10(-7) mol/L) of 2 h- shocked lymphatics and the Delta CF (1 x 10(-7) mol/L, 3 x 10(-7) mol/L), Delta TI (1 x 10(-7) mol/L) and Delta CA (1 x 10(-7) mol/L) of 3 h- shocked lymphatics were all significantly reduced when compared with control group.

CONCLUSIONThe reactivity of lymphatics to SP presented a biphasic change during the process of HS: increase in early phase and decline in later stage.

Animals ; Lymphatic Vessels ; physiopathology ; Male ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology ; Substance P ; analysis ; Thoracic Duct ; physiopathology