Objective To evaluate the antimicrobial susceptibility of bacteria cultured on a Poloxamer 407(P407) thermosensi-tive in-situ gel .Methods The modified Kirby Bauer(K-B) disc diffusion method was adopted to determine the drug susceptibility of 9 kinds of bacterium isolated on the P407 and the Mueller Hinton(MH)agar culture medium, at the same time the diffusion rates of different antibacterial drugs on the P407 and the MH agar culture medium were compared;the sodium dodecylsulfate polyacrylam-ide gel electrophoresis(SDS-PAGE) was adopted to analyze the outer membrane proteins (OMPs) expression of Pseudomonas aeruginosa on these two kinds of culture medium .Results In the antimicrobial susceptibility tests, the inhibition zones on the P407 gel were found to be generally smaller than those on the MH agar.The difference in the diffusion rates of the antibacterial drugs on these two kinds of culture medium had no statistically significant difference (P> 0.05);the OMPs expression of Pseudomonas aeruginosa was similar to its growth expression on the P407 culture medium ,but different from its expression on the MH agar .Con-clusion P407 as an excipient of bacteria culture medium can be used in the screening test of the biofilm antimicrobial susceptibility .

Objective To investigate the diagnostic and therapeutic methods of dural arteriovenuoe fistula in carotid cavernous sinus region.Methods Twenty five cases of clinical diagnosis of dural arteriovenous fistula of carotid cavernous sinus region were identified by whole cerebral angiography with seldingers technique.Among them 13 cases underwent endovascular embolized therapy due to large amount of shunt,and another 12 cases were undertaken conservative method of compressing carotid artery of affected side on basis of lesser amount of shunt with a few number of feeding arteries.Results Conditions of the 25 cases revealed by DSA were selected for the proper indications and then underwent the formentioned therapy with all successful achievement.Conclusion Diagnosis of dural arteriovenous fistula of carotid cavernous sinus region is not difficult.Emdovascular embolization is very effective,but the conservative compressing method of carotid artery of the affected side is also an efficient therapy.(J Intervent Radiol,2006,15:195-197)

Breast cancer is the principal cause of death in malig-nancy women , usually treated with the combination of surgery , chemotherapy , radiotherapy and endocrinotherapy .With the de-velopment of cell biology , molecular biology , immunology, im-munotherapy becomes a new field of breast cancer treatment .In this review, we discuss new findings in breast cancer immuno-therapy , including recent successes with bispecific antibodies and immune checkpoint blockade .We also discuss therapeutic cancer vaccines and highlight several additional immunotherapy modalities in early stages of development .

Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.

The hospital observed the principles of ensuring medical safety,shared risks and responsibilities in its attempt to build the risk mortgage mechanism in the hospital.The mechanism is characteristic of setting up the scope and standards for the risk mortgage,defining the use and refund cycle of the risk mortgage,along with a series of rewards and penalties.This design aims at minimizing cases of medical disputes (malpractice) for better medical safety.

Aim To investigate the protective effects of hydrogen sulfide ( H2 S) on focal cerebral ischemia/reperfusion injury in rats and the possible mechanisms. Methods Male Sprague Dawley rats were divided into three groups randomly: sham-operated group, cerebral ischemia/ reperfusion ( I/R) group and sodium hydro-sulfide ( NaHS ) + I/R group. The left temporary middle cerebral artery occlusion ( MCAO ) model was established by the line-embolism method. After rats were suffered 2h/24h ischemia/reperfusion stress, the mortality rate was evaluated, and the nervous function-al defect degree was evaluated by Longe scoring, the volumes of cerebral infarction was evaluated by 2 ,3 ,5-triphenyltetrazolium chloride ( TTC) staining, and the expression of P2X7 receptor protein in brain tissue was detected by the immunofluorescence method. Results The mortality rate in NaHS + I/R rats ( 29.41%) was obviously lower than those of I/R group ( 42 . 86%) . The nervous defect scores in NaHS + I/R rats were significant lower than those of I/R group ( P <0.05 ) . The volumes of cerebral infarction in NaHS +I/R group (21.88% ±3.53%) were significant lower than those of I/R group ( 36.71% ±3.73%) ( P <0.01 ) . The results of immunofluorescence showed that the positive expression cells of P2X7 receptor protein in cerebral cortex and hippocampal CA1 area of I/R group were significantly higher than those of sham-op-erated group(P<0. 01). However, compared with I/R group, the positive expression cells of P2X7 receptor protein in cerebral cortex and hippocampal CA1 area of NaHS + I/R group were significantly decreased ( P<0. 01). Conclusions H2S exerts the neuroprotective effect on focal cerebral ischemia/reperfusion injury in rats, and the protective mechanism might be associated with down-regulating the expression of P2X7 receptor protein in brain tissue.

[ABSTRACT]AIM:ToexaminetheeffectsofhighconcentrationofextracellularATPonhumanneuroblastoma SH-SY5Y cell injury.METHODS: Cultured SH-SY5Y cells were grouped according to the concentrations of ATP and treatment time.The cell viability was detected by CCK-8 assay.The variation of autophagic vacuoles was observed with monodansylcadaverine staining .The cell apoptosis was analyzed by Hoechst 33258 staining.Meanwhile, apoptotic rate was detected by flow cytometry .The levels of caspase-3 and microtubule-associated protein 1 light chain 3-Ⅱ ( LC3-Ⅱ) were determined by Western blotting .RESULTS:Compared with control group , the survival rate of SH-SY5Y cells was signifi-cantly reduced by ATP at different concentrations (3, 6, 9, 12 and 15 mmol/L for 3 h) and different treatment time (1, 2, 3 and 6 h with 6 mmol/L ATP, peaking at 3 h).The autophagic vacuoles of SH-SY5Y cells were significantly increased at 1 h with ATP treatment , trended to decrease over time and returned to control level at 6 h.The protein expression of LC3-Ⅱwas significantly increased at 1 h with ATP treatment , which was consistent with the time points of increasing auto-phagic vacuoles .LC3-Ⅱexpression level gradually decreased at 2~3 h with ATP treatment , and returned to control level at 6 h.Compared with control group , the apoptotic rate and the expression level of caspase-3 were enhanced synchronously . The peak of apoptotic rate occurred at 3 h, and kept until 6 h.The level of cleaved caspase-3 expression peaked at 6 h. CONCLUSION:High concentration of extracellular ATP induces the autophagy and apoptosis of SH -SY5Y cells.The in-creased autophagy shows up , followed by the climax of apoptosis until 6 h.With the prolonged duration of ATP , apoptosis is the main process in the cells .

AIM: To examined the effects of hypoxic preconditioning ( HPC) on oxygen-glucose deprivation ( OGD)-induced PC12 cells, and to investigate its possible mechanisms of autophagy .METHODS: Cultured PC12 cells were randomly divided into control group , HPC group, 3-methyladenine (3-MA) group, HPC+OGD group, 3-MA+HPC+OGD group and OGD group .CCK-8 assay was used to detect the cell viability .The caspase-3 activity was also tested . TUNEL staining and flow cytometry were used to detect the cell apoptosis .The protein levels of apoptosis-related protein caspase-3 and autophagy-marked protein LC3-2 and beclin-1 were determined by Western blot .RESULTS:Compared with control group, the viability of PC12 cells was significantly reduced , and the activity of caspase-3 was significantly increased in OGD group.Compared with 3-MA+HPC+OGD group and OGD group , the viability of PC12 cells was significantly in-creased, and the activity of caspase-3 was significantly reduced in HPC +OGD group (P<0.05).The PC12 cell injury was apparent after OGD with a great increase in the apoptotic rate (P<0.05).Compared with OGD group, the apoptotic rate significantly decreased in HPC +OGD group ( P<0.05 ) .Compared with control group , the protein level of cleaved caspase-3 was significantly increased in OGD group ( P<0.05) .Compared with OGD group , the protein level of cleaved caspase-3 was significantly decreased , and the levels of LC3-2 and beclin-1 were significantly increased in HPC +OGD group (P<0.05).CONCLUSION:OGD decreases cell survival and induces apoptosis .Activation of cell autophagy may be the mechanism by which hypoxic preconditioning protects the PC 12 cells from OGD induced injury .

OBJECTIVE:To investigate the therapeutic efficacy of edaravone combined with citicoline sodium on acute cere-bral infarction and its effects on the levels of oxidative stress and inflammatory factors. METHODS:108 patients with acute cere-bral infarction were randomly divided into edaravone group(single group)and edaravone+citicoline sodium group(drug combina-tion group),with 54 cases in each group. Based on routine treatment,single group was given Edaravone injection 30 mg added in-to 100 ml 0.9% Sodium chloride injection intravenously,bid,used up within 30 min each time;drug combination group was addi-tionally given Citicoline sodium injection 0.5 g added into 250 ml 0.9% Sodium chloride injection intravenously,qd,on the basis of single group. Treatment course of 2 groups lasted for 2 weeks. NIHSS,HDS,Barthel index,oxidant stress indicator and inflam-matory factors were compared between 2 groups before and after treatment. RESULTS:After treatment,the effective rate of NI-HSS in drug combination group was 81.48%,which was significantly higher than single group(53.70%),with statistical signifi-cance (χ2=9.511,P=0.002). HDS score and Barthel index of 2 groups were significantly increased after treatment,especially in drug combination group,with statistical significance(P<0.05). Compared with before treatment,contents of MDA and ET-1 in 2 groups were decreased significantly,while SOD activity and NO content were increased significantly;the inflammatory cytokines IL-6,IL-8,IL-12 and IL-16,TNF-α were all decreased gradually,with statistical significance (P<0.05);the improvement of each indicator in drug combination group was more significant than single group,with statistical significance(P<0.05). CONCLU-SIONS:Edaravone combined with citicoline sodium show good therapeutic efficacy in the treatment of acute cerebral infarction, can decrease the levels of oxidative stress and inflammation and promote the recovery of the neurological function and the daily liv-ing ability.

Objective To study the relationship between the methylation status of CPG islands in MEG3 gene promoter region of epithelial ovarian cancer and its clinical and pathological features. Methods The promoter methylation status was evaluated by MSP (methylation-specific polymerase chain reaction ) in 47 cases of ovarian cancer tissue and 15 cases of normal control. Results The methylation ratio (42.6%) of the MEG3 genes in the ovarian cancer was statistically significantly higher (P = 0.035 ) than that (13.3%) in the normal control. The methyation rate of the group with an age > 60 years old was slightly higher than that of the group with an age≤60 years old, without statistically significant (P > 0.05), so was observed in ovarian cancers of stage Ⅰ andⅡ than that in stage Ⅲ and Ⅳ. There were also no significant differences in MEG3 gene methylation positive rate neither in different pathological grading nor in various ovarian cancer tissues (P > 0.05). Conclusion Abnormal methylation in MEG3 gene may be associated with epithelial ovarian cancer , but no relation to its clinical pathology.