OBJECTIVE To investigate antimicrobial resistance of Enterobacter cloacae in our hospital,for guiding the treatment of these infections in clinical practice.METHODS Bacterial susceptibility testing was done by Kirby-Bauer method and bacteria were identified by VITEK-32.Retrospective analysis of the drug resistance was done to the E.cloacae isolated from our hospital in the recent 3 years.RESULTS A total of 364 E.cloacae strains were isolated,which mainly isolated from sputum.They were resistant to 13 types of antibacterial agents but the resistance rate to imipenem was 3.30%.The resistant rate to third and fourth generation cepholosporins(excepting cefoperazone/sulbactam) and quinolones was more than 30% and to ampicillin and cefazolin was more than 94.00%.CONCLUSIONS E.cloacae is severely resistant to cepholosporins and quinolones.More attention should be paid to the surveillance of such strains.Imipenem may be considered for use in critical patients.

Objective To investigate the feasibility of video-assisted thoracoscopic sympathetic trunk clipping in the treatment of craniofacial hyperhidrosis. Methods A total of 10 patients were operated on under general anesthesia with double-lumen endotracheal intubation. The patients were placed in lateral recumbent position with one-lung ventilation. A 7 mm trocar and a 4.5 mm trocar were inserted at the 2~3 intercostal space on the midaxillary line and at the 4~5 intercostal space on the posterior axillary line, respectively, to introduce surgical instruments and thoracoscopic camera. Alongside the sympathetic chain, the sympathetic nerve trunk immediately below the second costal margin was blocked with small-sized titanium clips. Then the lung was inflated and the incision sutured. Afterwards, the procedure in the contralateral hemithorax was performed using the same method. Results The operating time was 55~130 min (mean, 110 min). Symptoms of craniofacial hyperhidrosis disappeared in the 10 patients, all of who were satisfied with curative results. The postoperative hospital stay was 2~3 days. Neither Horner’s syndrome nor other serious complications were observed. Seven of the patients developed slight compensation hyperhidrosis in their chest, abdomen, back or legs. All the patients had normally returned to work and physical exercises in 7~10 days. Postoperative follow-up for 1~9 months (mean,6.3 months) in all the patients found no recurrence. Conclusions Video-assisted thoracoscopic block of sympathetic trunk below the second costal margin for craniofacial hyperhidrosis is safe and effective.

Objective To evaluate the expression of Sp3 gene of peripheral blood mononuclear cells (PBMC) in multiple sclerosis (MS) patients in Guizhou and the relationship between Sp3 gene expression and immunological function. Methods Two pairs of primers were used to amplify cDNAs generated from 31 MS patients and 30 healthy controls. The serum levels of sIL-2R were measured in 27 patients with MS and 30 healthy controls by sandwiched ELISA. Results The deficient expression of Sp3 gene in MS patients was significantly higher than that in control (41.9% ( 12/31 ) vs 6. 7% (2/30) ,x2 =7. 133 ,P =0. 008). The sIL-2R levels in MS patients were significantly higher than those in control (( 2788.5 ± 1079. 8 ), ( 1270. 7 ± 489. 4) μg/L, t = 6. 170, P = 0. 001 ). The concentration of sIL-2R in MS with negative ((3364.0 ± 1252.3) μg/L) and positive((2450.0 ± 827.0) μg/L) expression of Sp3 gene were significantly increased compared with control (F = 32. 059, P < 0. 05 ). The sIL-2R levels were significantly rising in MS patients with negative expression of Sp3 gene compared with MS patients with positive expression of Sp3 gene ( q = 4. 213, P < 0. 05 ). Conclusions A remarkable deficient expression of Sp3 gene in PBMC has been found in MS patients in Guizhou. sIL-2R may take part in the process of MS. The expression of Sp3 gene is not affected by immune state, however, MS patients with Sp3 deficient expression tend to have a more serious impairment in immunological functions.

Optimization of the fermentation condition for human apolipoproteinA-I expression in recombinant Escherichia coli was investigated. The recombinant plasmid pBV220-ApoA-I was transformed respectively into different E.coli hosts such as JM109, BL21(DE3),DH5?, BMH7118,and TG1. The best host E.coli was DH5? in which the recombinant ApoA-I expression percentage was 21.2% corresponding to that in BL21(DE3) in flask shaker cultivation,while the ApoA-I expressed percentage in E.coli TG1 was 11%.Fed-batch cultivation was performed in FMG-5L fermentor,the optimum fermentation cultivation conditions were as following :optimum pH value was 7.0 in growth phase and 7.4 in the expression phase. The initial glucose concentration in batch phase was 3 g?L -1.The optimum C/N ratio was 2∶1.The recombinant ApoA-I reached about 40% of the total protein, and concentration of ApoA-I was 2.86 g?L -1.

The temperature effect on the recombinant protein production formation was investigated in present study. The culture temperature of growth phase is 30℃, and the culture temperature of induction phase was arranged according to three modes. Hign cell-density and high expression culture of E.coli to product recombinant human apolipoprotein A-I Milano by two temperature-shifted induction . Two temperature-shifted induction was carried out high density and high expression recombinant human ApoA-1 Milano. The recombinant protein ApoA-I Milano reached 4.8 g?L -1 with the final cell density of OD 600 150. And the two temperature-shifted induction avoided the acetic acid successfully to the influence of the high density and high expression. Two temperature-shifted induction was viable in high density culture and high expression of heterogenous protein in recombination E.coli.The sduty provides a basic work for production of recombinant ApoA-I Milano in scale.

Objective To investigated the effects of combined arsenic trioxide(ATO) and resveratrol(Res)on the viability of NB4 human leukemia cells. Methods NB4 human leukemia cell was used in this experiment.Cells were cultured in ATO (0,0.1875,0.3750,0.7500, 1.1250, 1.5000,2.2500,3.0000,5.0000 μmol/L) and Res (0, 1.5625,3.1250,6.2500, 12.5000, 18.7500,25.0000,37.5000,50.0000 μmol/L). Cell viabilities were measured by MTT in different treatment groups. Half inhibitory concentration(IC50) was calculated. The ratio of concentration of ATO and Res 1.5∶ 18,1.5∶ 25,1.5∶ 35 was added to cells, and the combination index(CI) was calculated. The level of ROS in control, ATO( 1.5000 μmol/L), Res(25.0000 μmol/L) and ATO(0.9000 μmol/L) + Res( 12.5000μmol/L) groups was measured by chemiluminescence assay. Results ①ATO( ≥0.7500 μmol/L) reduced the viability of NB4 cells in a concentration-dependent manner(P ＜ 0.05 ), and IC50 was (1.78 ± 0.11 )μmol/L. ②)Res (≥18.7500 μ mol/L) dose-dependently decreased the viability of NB4 cells (P ＜ 0.05 ), and IC50 was ( 18.71 ±0.18)μ mol/L. ③Combination of ATO and Res showed an antagonistic effect on NB4 cells viability. ④The ROS in Res group( 1670.55 ± 13.97) was significantly lower than that in control group(2345.88 ± 14.48,P ＜ 0.05). The ROS in ATO group (3092.42 ± 94.84) was significantly higher than that in control group(P ＜ 0.05). The ROS in ATO + Res group (1860.27 ± 15.99) was significantly lower than that in ATO group(P ＜ 0.05). Conclusions NB4 cell survival rate can be decreased by ATO and Res. The combination of arsenic trioxide and Res presents an antagonistic effect on NB4 cell viability, in part by reducing intracellular ROS formation.

AIMTo prepare the breviscapine liposomes and study the pharmacokinetics of breviscapine liposomes in Beagle dogs.

METHODSThe cross-over design (two periods) was employed. Six Beagle dogs were administrated a single intravenous dosage of 28 mg of breviscapine liposomes and reference preparation, respectively, scutellarin in plasma of 6 dogs at different sampling time was determined by RP-HPLC. The pharmacokinetic parameters were calculated by 3P97 program and compared by statistic analysis.

RESULTSThe mean concentration-time curves of breviscapine liposomes and reference preparation were both fitted to two-compartment model with the main pharmacokinetic parameters as follows: T 1/2 alpha were (4.4 +/- 0.7) min and (1.8 +/- 1.3) min respectively; T 1/2 beta were (55 +/- 27) min and (28 +/- 23) min respectively; V(c) were (1 580 +/- 265) mL and (2 460 +/- 2 200) mL respectively; CL(s) were (88 +/- 10) mL x min(-1) and (324 +/- 69) mL x min(-1) respectively; and AUC(0-720) were (363 +/- 42) microg x min x mL(-1) and (102 +/- 19) microg x min x mL(-1) respectively. The T 1/2 alpha, CL(s) and AUC(0-720) of breviscapine liposomes all had significant difference from those of reference preparation, after the data were examined by a one-way analysis of variance (ANOVA).

CONCLUSIONCompared with the reference preparation, breviscapine liposomes had a much more higher concentration in plasma and contained characteristic of sustained-release, which ameliorated the pharmacokinetic properties of scutellarin.

Animals ; Apigenin ; blood ; Area Under Curve ; Brain ; metabolism ; Cross-Over Studies ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; Drug Stability ; Erigeron ; chemistry ; Female ; Flavonoids ; administration & dosage ; isolation & purification ; pharmacokinetics ; Glucuronates ; blood ; Injections, Intravenous ; Liposomes ; Male ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the effect of hepatocyte growth factor (HGF) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT) and related mechanism in acute lymphoblastic leukemia (ALL) mice.

METHODSTwenty nude mice were randomly divided into control (group A) and test (group B) groups for monitoring relapse, and 20 BALB/c mice into control (group C) and test (group D) groups for GVHD. HGF as injected from day 0 to day 7 after BMT for groups B and D, while PBS for A and C. CD4(+) and CD8(+) T cell were evaluated by flow cytometry. The survival of mice after BMT was recorded. The level of tumor necrosis factor-alpha (TNF-alpha) was evaluated by ELISA.

RESULTSThe median past-BMT survival were 7.00 +/- 1.58, 9.00 +/- 1.58, 11.00 +/- 3.95 and 24.00 +/- 13.44 days for groups A, B, C, D, respectively, being prolonged in group D. HGF could decrease the quantity of CD4(+) T cells [group D (10.39 +/- 1.15)% vs group C (13.50 +/- 1.80)%, P < 0.01] and increase CD8(+) T cell [group D (12.25 +/- 2.85)% vs group C (6.12 +/- 1.99)%, P < 0.01], decrease the level of TNF-alpha in transplanted ALL mice [group D (112.10 +/- 18.99) pg/ml vs group C (143.90 +/- 25.35) pg/ml, P < 0.01] and reduce the degree of GVHD.

CONCLUSIONHGF could alleviate post-allo-BMT GVHD but retain GVL effect.

Animals ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; surgery ; Random Allocation ; Transplantation, Homologous

Objective:The prokaryotic expression vector of human anti-Siglec-9 Fab fragment antibody was constructed and purified,while was identified. Methods:The variable and conserved regions of heavy chain and light chain were obtained by polymerase chain reaction respectively(PCR),which was combined by overlap extension PCR and was digested with restriction enzyme,and then it was transformed into Escherichia coli BL21 and was purified by His-trap Lambda Fab column and AKTA system. SDS-PAGE,ELISA and Western blot were used for the identification of human anti-Siglec-9 Fab fragment antibody. The effect of human anti-Siglec-9 Fab fragment antibody on regulating the mRNA expression of TNF-α,IL-1,IL-6,IL-8 was detected by real-time PCR. Results:Successfully obtained the chains of heavy and light, while constructed an activation human anti-Siglec-9 Fab fragment antibody which could specifically bind to Siglec-9 protein. The human anti-Siglec-9 Fab fragment antibody could specifically bind to Siglec-9 was confirmed by SDS-PAGE,ELISA and Western blot. The human anti-Siglec-9 Fab fragment antibody inhibited the mRNA expression of TNF-α,IL-1, IL-6,IL-8. Conclusion:Successful prokaryotic expression,purification,character analysis,and suppressed the mRNA expression of in-flammatory cytokines with the human anti-Siglec-9 Fab fragment antibody and lay the biology foundation for the further study.

OBJECTIVE To illuminate the adenoid bacteria distribution in children with adenoid hypertrophy. METHODS PubMed, Embash, Medline, CNKI, VIP Information and Wanfang data were searched for studies on the adenoid bacteria distribution and adenoid hypertrophy. Random effects meta-analysis was used to pool data. RESULTS Nine studies were included in this meta analysis. The pooled detection rates of haemophilus influenza, staphylococcus aureus and streptococcus pneumonia were 0.21 (95%CI, 0.09-0.32), 0.14 (95%CI, 0.09-0.20) and 0.15 (95%CI , 0.08-0.22) respectively. CONCLUSION Haemophilus influenzae, staphylococcus aureus, and streptococcus pneumoniae are three main kinds of pathogenic bacteria of adenoid hypertrophy in children.