OBJECTIVE To investigate the disinfectant-resistant gene qacE△1-sul1 in Pseudomonas aeruginosa isolated from burned patients. METHODS GNS-448 and K-B tests were performed to detect the susceptibility of 19 kinds of antimicrobial agents against these strains. Genotype was analyzed by polymerase chain reaction(PCR) and verified by DNA sequencing. RESULTS The 32 strains isolated were all resistant to ampicillin,cefuroxime,cefoxitin,SMZ/TPM. The sensitive rates to amikacin and gentamicin were 68.0%,and 46.9%,respectively,the resistant rates to imipenem and meropenem were 68.8% and 59.4%,respectively. The positive rate of gene qacE△1-sul1 was 50.0%. CONCLUSIONS The resistance of P. aeruginosa isolated from burned patients is a serious issue.There is high positive percentage of qacE△1-sul1 gene in P. aeruginosa isolated from burned patients.

OBJECTIVE To investigate the 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes in Pseudomonas aeruginosa isolated from burned patients. METHODS GNS-448 and K-B tests were performed to detect the susceptibility to 19 kinds of antimicrobial agents against these strains. 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes were amplified by polymerase chain reaction (PCR) and verified by DNA sequencing. RESULTS The 32 isolated strains were all resistant to ampicillin,cefuroxime,cefoxitin,SMZ-TMP,The sensitive rates to amikacin and gentamicin were 68% and 46.9%,respectively. The resistant rates to imipenem and meropenem were 68.8% and 59.4%,respectively. The 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes including aac(6')-Ⅰb,aac(6')-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and rmtB were found and positive rates were 9.4%,3.1%,28.1%,25.0% and 3.1%,respectively. A novel subtype of aac(6')-Ⅰb was reported firstly. CONCLUSIONS There are high positive percentage of 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes in P. aeruginosa isolated from burned patients. P. aeruginosa resistance to aminoglycoside relates to the existence of 16S rRNA methylase gene and aminoglycoside-modifying enzyme genes.

Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.

Objective To observe the effects of different levels of stroke volume variation (SVV)directed fluid therapy on prognosis in elderly patients.Methods One hundred and ten patients (male 67 cases,female 43 cases,aged 65-80 years,ASA grade Ⅰ or Ⅱ)undergoing abdominal sur-geries were randomized into three groups on the basis of SVV level:1 1%≤SVV≤14% (group H,n=37),7%≤SVV≤10% (group M,n = 37),3% ≤ SVV≤ 6% (group L,n = 36).Each group ac-cepted different fluid therapy strategy.Perioperative intake and outtake volume were recorded.Progno-sis indicators included gastrointestinal function recovery,post and total length of hospital stay and so on were collected.Postoperative complications were also recorded.Results Along with the level of SVV dropped,The volume of colloid and crystalloid among groups were significantly increased (P <0.05).There were no remarkable difference in the time interval of activity off bed,liquid diet,gastrointestinal decompres-sion and catheterization.Group L had a delayed exhaust time compared with group H (P <0.05).Patients in the group M had a shorter time to have semi-liquid diet.The transition of diet was also shortest in the group M.But these differences were not statistically significant.The group M also had a shorter time in first-degree or above nursing time and post and total length of hospital stay compared with group L (P <0.05).The incidence of postoperative infection in group L was higher than other two groups.Incidence of anastomosis leakage in group H was higher than that in other two groups (P <0.05).Conditions of compli-cations in group L were more severe and complicated compared with other two groups.Conclusion 3%≤SVV≤6% may increase the risk of postoperative infection and prolong the time of hospitalization.Patients in the level of 7%≤SVV≤10% may have a better postoperative recovery.This level could be a better goal of perioperative fluid therapy in elderly patients.

OBJECTIVE: To investigate the influence of commercial peritoneal dialysate (CDS) on function of macrophage. METHODS: Cultured peritoneal macrophages were divided into two experimental groups and their controls.(1) Macrophages were cultured in conditioned culture medium containing 50%CDS (0.139 mol/L glucose) for 24 h. (2)Macrophage were exposed to CDS containing 0.139mol/L glucose for 10, 30 and 60 min respectively, and then cultured in CDS-free medium for 24 h. The nitric oxide (NO) production and MTT in two groups were measured. In each control group, CDS was replaced by same amount of culture medium. RESULTS: NO production and MTT reduction ability (related to cell viability) of experimental groups were remarkably lower than those of controls and the NO production and MTT reduction in 60min CDS-exposed group were lower than that of 10 min and 30 min CDS-exposed group. CONCLUSION: Dialysate may have detrimental effects on viability and other function of macrophage and these effects may related to time length of CDS exposure.

Based on single factor tests,the optimum vacuum belt drying conditions of Qibai Pingfei granule were obtained through Box-Benhnken central combination design and RSM. In this study, drying time, drying temperature and extract density were chosen as independent variables, while transferring rate ginsenoside Rg₁, Re, Rb₁and astragaloside IV were taken as dependent variables. The optimum parameters are as follows: drying time of 112 min, drying temperature of 87 °C and extract density of 1.30 g · mL⁻¹. At the optimum condition, transferring rate ginsenoside Rg₁+ Re, Rb₁and astragaloside IV were 88.01%, 87.31%, 84. 34%. Above all, the optimum processing parameters of vacuum belt drying of Qibai Pingfei granule is reasonable and feasible, which can provide reliable basis for production.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Temperature ; Vacuum

To study the anti-proliferation effect of lambda-carrageenan oligosaccharides (lambda-CO) on human umbilical vein endothelial cells (HUVECs) and expression of apoptotic relevant genes, the influence of lambda-CO on HUVECs proliferation was measured by MTT assay; apoptotic rate, cell cycle distribution and the level of active caspase-3 of HUVECs were analyzed using flow cytometry; the mRNA level of apoptosis related genes was determined by RT-PCR. At a high concentration of 1 mg x mL(-1), lambda-CO significantly inhibited the endothelial cell proliferation. Annexin-V FITC/PI double stain assay showed that when treated with 0, 0.8, 1 mg x mL(-1) of lambda-CO for 24 h, cell apoptotic rates were (1.67 +/- 1.6)%, (11.48 +/- 2.4)% and (13.81 +/- 2.2)%, respectively, when treated for 48 h, cell apoptotic rates were (2.02 +/- 2.3)%, (13.84 +/- 1.9)% and (38.72 +/- 2.5)%, respectively, cell cycle assay showed the decrease of cells in G0/G1 phase, and increase in S phase. Furthermore, we observed the level of active caspase-3 increased in a dose-dependent manner at 24 th and 48 th. RT-PCR results indicated that mRNA of TNFalpha, p53, caspase-8 and caspase-3 in cells increased after treated with lambda-CO. lambda-CO induce apoptosis of HUVECs in a dose-dependent way and arrests cells at S phase, which mainly due to the up-regulation of apoptotic genes such as TNFalpha, p53, caspase-8, caspase-3 and increase the level of active caspase-3.
Angiogenesis Inhibitors ; pharmacology ; Apoptosis ; drug effects ; Carrageenan ; pharmacology ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; biosynthesis ; genetics ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Humans ; Oligosaccharides ; pharmacology ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Umbilical Veins ; cytology

2-(4-Methylthiazol-5-yl) ethyl nitrate hydrochloride (W1302) is a nitro containing derivative of clomethiazole, which is a novel neuroprotective agent with both carbon monoxide (NO) donor and weak γ-aminobutyric acid type A (GABA_A) receptor allosteric regulatory excitatory effect. The current study used a rat model of transient middle cerebral arteryocclusion (tMCAO) brain injury to evaluate the therapeutic effect of W1302 on ischemic stroke and explore its potential mechanisms of action. This experiment has been reviewed and approved by the Laboratory Animal Management and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences (ethical review forms No. 620, 632, 5013). The results showed that gavage administration of 1, 3, and 10 mg·kg^-1 of W1302 can significantly reduce the volume of cerebral infarction in rats with ischemia for 2 h and reperfusion for 24 h, and the therapeutic effect was better than that of 200 mg·kg^-1 of DL-3n-butylphthalide. W1302 significantly increased NO levels in blood and brain tissue. It increased cerebral blood flow and brain adenosine triphosphate (ATP) content after reperfusion, as well as, inhibited the expressions of inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue. The time window of W1302 was between 120-180 min after ischemia. These research results demonstrated that W1302 could increase NO release, dilate blood vessels, and increase cerebral blood flow, improve energy supply to brain tissue and increase ATP levels, and inhibit neuro-inflammation, which playing the protective roles in tMCAO stroke model rats. This provides theoretical support for the clinical application of W1302 in the treatment of ischemic stroke.

OBJECTIVETo investigate the role of oxidative stress in the pathogenesis of esophageal mucosa injury in children with reflux esophagitis (RE).

METHODSEsophageal mucosal samples from 36 children with RE (7 months to 16 years of age) were obtained by gastroscopy. The parameters of oxidative stress, including the contents of malondialdehyde (MDA), glutathione (GSH) and nitric oxide (NO) and total superoxide dismutase (T-SOD) activity in the esophageal mucosa as well as the protein content of the esophageal mucosa, were measured. Twenty children (3 to 16 years of age) without esophageal mucosal injury by gastroscopy served as controls.

RESULTSThere was no significant difference in the protein content of the esophageal mucosa between the RE and the control groups. The content of MDA in the RE group (15.36+/- 16.67 nmol/mg) was significantly higher than that in the control group (7.51+/- 6.17 nmol/mg) (P<0.01). The activity of T-SOD in the RE group (30.43+/- 35.09 U/mg) was statistically lower than that in the control group (56.34+/- 51.73 U/mg) (P<0.05). No significant differences were observed in GSH and NO contents between the two groups.

CONCLUSIONSThe MDA content increases and the SOD content decreases in the esophageal mucosa in children with RE. This suggests that oxidative stress seems to be an important mediator in generation of esophageal mucosal injury.

Adolescent ; Child ; Child, Preschool ; Esophagitis, Peptic ; complications ; metabolism ; Esophagus ; metabolism ; Female ; Glutathione ; metabolism ; Humans ; Infant ; Male ; Malondialdehyde ; analysis ; Mucous Membrane ; metabolism ; Nitric Oxide ; biosynthesis ; Oxidative Stress ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the relationship between the drug resistance of Pseudomonas aeruginosa (PA) isolated from burn patients wounds and its mobile genetic elements, including plasmid, transposon, and integron.

METHODSThirty-two strains of PA were isolated from wounds exudate of hospitalized burn patients in Ningbo No. 2 Hospital. PA drug sensitivity was determined using GNS-448 drug sensitivity card and K-B tests. The genetic markers of plasmid, transposon and integron including traA, traF, tnpA, tnpU, merA, int I 1 were amplified by PCR and verified by gene sequencing.

RESULTSDrug resistant rate of 32 PA strains to gentamicin, amikacin, cefoperazone/sulbactam, ciprofloxacin was 43.7%, 32.0%, 46.8%, 49.9%, respectively. PA drug resistant rates to piperacillin, cefotaxime, ceftazidime, cefepime, aztreonam, piperacillin/tazobactam, levofloxacin, imipenem and meropenem were all above 56.0%. Seventeen out of 32 PA strains were found to carry transposon and (or) integron genetic markers. One strain was positive for both tnpA and merA, 8 strains were positive for both merA and int I 1, 1 strain was only positive for tnpA, 2 strains were only positive for merA, and 5 strains were positive for int I 1 only.

CONCLUSIONSPA isolated from burn wounds of hospitalized patients in Ningbo No. 2 Hospital is seriously drug resistant, which may relate with its high positive rate of mobile genetic elements of transposon and (or) integron.

Anti-Bacterial Agents ; pharmacology ; Burns ; microbiology ; DNA Transposable Elements ; Drug Resistance, Multiple, Bacterial ; genetics ; Humans ; Integrons ; Microbial Sensitivity Tests ; Plasmids ; Pseudomonas aeruginosa ; drug effects ; genetics ; isolation & purification