Objective:To optimize the ethanol extraction technology for Xiaoluo granules. Methods:The extraction technology for the ethanol dissolvable ingredients in 10 kinds of medicinal materials was optimized by orthogonal experiment. The weighted sum of ba-icalin content and the dry extract rate as the index, 4 influencing factors with 3 levels each were optimized by L9 (34 ) orthogonal table. Results:The optimal ethanol extraction conditions were as follows:the medicinal material was soaked by 10-fold 50% ethanol solution for 45min, and then extracted by heating reflux for 3 times with 1. 5h each time. Conclusion: The method can effectively extract the ethanol dissolvable ingredients in 10 kinds of medicinal materials, which can be used as the ethanol extraction technology for Xiaoluo granules.

Objective To investigate the influence of poly(ADP-ribose)polymerase inhibitor PJ34 on blood brain barrier(BBB)in rats with cere-bral ischemia-reperfusion injury. Methods Rat model of cerebral ischemia-reperfusion injury was established by the middle cerebral artery occlu-sion. A total of 135 SD rats were randomly divided into 3 groups:sham-operated group(sham group),ischemia-reperfusion group(IR group)and PJ34 group(PJ34 group). 45 animals in each group were then equally divided into subgroups and the rats were sacrificed at 6 h,24 h,48 h after re-perfusion,respectively. BBB permeability was evaluated by detection of extravasated Evans blue(EB). The expression of tumor necrosis factorα(TNF-α)and matrix metalloproteinase 9(MMP-9)activity were measured by immunohistochemistry and western blot at different time points. Re?sults Compared with sham group,the contents of EB and the expressions of TNF-αand MMP-9 in IR group were increased significantly(P<0.05). Compared with IR group,the contents of EB and the expressions of TNF-αand MMP-9 in PJ34 group were markedly decreased at the same time point(P<0.05). Conclusion The present study provided in vivo evidence that PARP inhibitor PJ34 can protect against cerebral ischemia re-perfusion injury,and the mechanism might be related to maintaining the stability of blood-brain barrier by suppressing the expression of TNF-αand MMP-9 in ischemic cortex.

Objective To evaluate different tigecycline susceptibility testing methods for A.baumannii.Methods Thirty carbapenem resistant A.baumannii (CRAB) and 30 carbapenem sensitive A.baumannii (CSAB) isolates were randomly collected from 30 hospitals during January to December in 2010 in China retrospectively.MIC and inhibitory zone diameters for tigecyclinc were determined by the susceptibility testing methods such as broth microdilution (BMD),agar dilution,E test,MIC Test Strip (MTS),Vitek2.Data were analyzed by comparing the results from each method to those produced by the reference BMD method.The effects of two different susceptibility test media (M-H and ISO-Sensitest Agar) on the MIC of tigecycline were also analyzed.Results For CSAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:0.125/0.25 mg/L,0.125/0.25 mg/L,0.5/1 mg/L,0.125/0.25 mg/L and 0.5/0.5 mg/L.Compared with BMD method,the categorical agreement rates (CA) of each method were ≥90％,and produced no very major errors (VME) by Food and Drug Administration (FDA)/ European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints.For CRAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:2/4 mg/L,4/4 mg/L,4/4 mg/L,1/2 mg/L and 2/4 mg/L.Compared with BMD method,MTS produced 3.3％ (1/30)/6.7％ (2/30) VME(FDA/EUCAST breakpoints),and no method CA was ≥90％.The CA between disk diffusion and BMD results were higher by using the criteria of Jones than FDA breakpoints,but only 66.7％ (20/30) were observed in CRAB isolates,and produced no VME.The MIC of tigecycline determined using M-H agar were usually higher than those using ISO-sensitest Agar.Conclusions Agar dilution,E test,Vitek 2 and disk diffusion appear not to be a suitable method for routine susceptibility testing of tigecycline for CRAB strains.Tigecycline intermediate or resistant results determined by these methods require confirmation by BMD,and MTS results also need to be interpreted with caution.

Objective To investigate the effects of PJ34,a poly ADP-ribose polymerase(PARP)inhibitor,on the expression of matrix metallopro-teinases-9( MMP-9 )and Claudin-5 in ischemic cortex of rats with focal cerebral ischemia-reperfusion(I/R)injury. Methods The focal cerebral ischemia-reperfusion model of middle cerebral artery occlusion(MCAO)was established by intraluminal suture . PJ34 was injected intraperitoneal-ly. Blood-brain barrier(BBB)permeability was quantitatively determined by Evans blue assay. Infarct volume changes were observed by 2,3,5-tri-phenyltetrazolium chloridedyeing(TTC)staining. The expression of the MMP-9 and Claudin-5 in rats of cerebral cortex were measured by immuno-histochemistry assay and western blot analysis . Results Compared with sham group,the expression of MMP-9,the contents of EB and infarct vol-ume increased progressively over time after I/R,and reached maximum levels at 48 h(P<0.05);The expression of Claudin-5,the contents of EB and infarct volume reduced significantly over time after I/R,and reached the minimum levels at 48 h in model group(P<0.05). Compared with model group,the expression of MMP-9,the contents of EB and infarct volume was reduced significantly and the expression of Claudin-5,the con-tents of EB and infarct volume was increased at the same time point in PJ34 group(P<0.05). Conclusion PJ34 maintained the blood-brain barri-er permeability by inhibiting the expression of MMP-9,and increasing the expression of Claudin-5,which had neuroprotection on cerebral ischemia-reperfusion injury.

Objective To optimize ethonal extraction technology for no sugar runfei granules.Methods Ethanol was used as a solvent to extract the ethanol dissolvable ingredients in 9 kinds of medicinal materials.The formula L9 (34 ) table was used to examine the effects of 4 factors and 3 levels.The weighted sum of baicalin content and the dried extract quantity was used as quantitative index.Results The maximize optimize condition for extraction of ethanol dissolvable ingredients was as follows:9 kinds of medicinal materials, add of 10-fold 70% ethanol solution, soaked for 45 min, and extracted by heating reflux for 3 times,1.5 h each time.Conclusion The method can maximize extraction of ethanol dissolvable effective ingredents in 9 kinds of medicinal materials and can be used as ethonal extraction technologgary of no sugar runfei granules.

Objective To analyze results of atopy patch test (APT) with dust mite allergens at different concentrations in patients with atopic dermatitis (AD),and to evaluate the diagnostic value of APT.Methods Totally,85 patients with AD were enrolled into this study.All the patients underwent APT with 5 concentrations (3 000,5 000,7 000,10 000 and 12 000 pnu/g) of dust mite allergens,as well as skin prick test (SPT) with dust mite allergens.Dust mite allergens were obtained from two different manufacturers (group 1 and 2).Enzyme-linked immunosorbent assay (ELISA) was performed to detect allergen-specific immunoglobulin E (SIgE) in sera from these patients.The sensitivity,specificity and positive predictive value of APT,SPT and SIgE assay were compared,and the results of APT were compared among different concentrations of allergens and between allergens from different manufacturers.Results When SIgE assay served as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 79.41％,76.12％,and 64.29％ respectively for APT,73.53％,80.95％ and 67.57％ respectively for SPT with group 1 dust mite allergens,and 81.53％,77.78％ and 65.09％ respectively for APT,76.02％,79.85％ and 66.07％ respectively for SPT with group 2 allergens.When SPT was regarded as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 78.38％,77.42％,67.44％ respectively for APT,67.57％,84.21％,73.53％ respectively for SIgE assay with group 1 dust mite allergens,79.25％,80.63％ and 69.55％ respectively for APT,61.07％,82.54％ and 77.21％ respectively for SIgE assay with group 2 allergens.There were no significant differences in the sensitivity,specificity or positive predictive value of APT or SPT between the two groups of allergens.The positive rate of APT was 8.24％,22.35％,29.41％,44.71％ and 41.18％ respectively with group 1 allergens at 3 000,5 000,7 000,10 000 and 12 000 pnu/g,and 3.53％,23.53％,31.76％,34.12％ and 35.29％ respectively with group 2 allergens.No significant differences were observed in the positive rate of APT between group 1 and 2 allergens at same concentrations (all P ＞ 0.05),but a significant difference was observed in that between different concentrations of group 1 or 2 allergens (both P＜ 0.05).The positive rate of APT increased with the increase of allergen concentrations,but stopped rising when the concentrations of group 1 and 2 allergens reached 7 000 pnu/g and 5 000 pnu/g respectively.Conclusions The sensitivity of APT is relatively high for the diagnosis of dust mite allergy.The positive rate of APT increased with the increase in allergen concentrations,but stopped rising when the concentrations reached a certain level.

Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.

BACKGROUND: Acetabular cup orientation using a standard radiograph of the pelvis is quite common method to assess artificial hip replacement nowadays. Non-standardization of pelvic orientation affected accuracy of measurement results, and it is difficult to compare. OBJECTIVE: To make sure how pelvis tilting affect the anteveration of the cup and to elevate clinical accuracy and compare study comparability. METHODS: Designed a simulated acetabular cup with serial concentric circles which pass through the same polars and represent anteveration of 0°, 10°, 20°, 30°, 40°, Loaded the simulated acetabular cup at an inclination of 35°, 40°, 45°, 50°, 55° to6 cadaver pelves, Made the pelves tilt around the frontal axis and sagittal axis with 5° each time in a scope of+30°. Takestandard radiograph of the pelvis accordingly. Radiograph was photographed end frontal angle of dip was measured. RESULTS AND CONCLUSION: Pelvic tilt of about 1° causes measuring errors of anteveration 0. 61 °-0. 73°. The anteveration decreased at both acetabular cups when pelvic posterior tilt and at the acetabular cup that near the X-ray source as pelvic lateral tilt. The anteveration rose at both acetabular cups when pelvic anterior tilt and at the acetabular cup that away from the X-ray source as pelvic lateral tilt. During clinical evaluation, pelvic orientation effects on measurement results should be considered.

ObjectiveTo investigate the risk factors of patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS) complicated with Yangming Fushi syndrome.Methods A prospective study was conducted. From August 2009 to July 2013, 206 patients with Yangming Fushi syndrome combined with ALI/ARDS were enrolled in the intensive care units ( ICUs ) of the following five hospitals: Tianjin Nankai Hospital, Dongzhimen Hospital Affiliated to Beijing Traditional Chinese Medicine University, General Hospital of Tianjin Medical University, the First Affiliated Hospital of Dalian Medical University, and Wuxi Third People's Hospital in Jiangsu Province. According to the mortality occurring in ICU, the patients were divided into death group and survival group. The gender, age, acute physiology and chronic health evaluation Ⅱ ( APACHE Ⅱ ) score within the first 24 hours after admission, the length of invasive mechanical ventilation, usage of vasoactive agents, ratio of operative intervention, the length of stay in ICU, application of continuous renal replacement therapy ( CRRT ), amount of blood transfusion, the level of C-reactive protein ( CRP ), lactulose/mannitol ( L/M ) ratio, the number of organs with dysfunction, oxygenation index ( PaO2/FiO2 ), levels of lactate and serum creatinine ( SCr ) of both groups were recorded. Logistic regression analysis was used to look for the independent risk factors of death of patients. Results There were 124 cases with severe acute pancreatitis ( SAP ), which was the most common disease with manifestation of Yangming Fushi syndrome combined with ALI/ARDS, accounting for 60.19% of all the patients. During the period of hospitalization in ICU, 171 patients survived and 35 died, and the mortality rate was 16.99%. The risk factors of the two groups were analyzed by variable analysis, and it was shown that compared with those in survival group, the age ( years: 57.26±16.23 vs. 48.07±13.48, t = 3.544, P = 0.000 ), APACHE Ⅱ score ( 20.83±9.73 vs. 12.61±6.63, t = 4.777, P = 0.000 ), the length of invasive mechanical ventilation ( days: 10.97±7.71 vs. 6.91±2.48, t = 2.555, P = 0.015 ) and the number of dysfunction organs ( 3.11±1.21 vs. 1.60±1.34, t = 6.222, P = 0.000 ) in death group were significantly higher. The level of PaO2/FiO2 [ mmHg ( 1 mmHg = 0.133 kPa ): 218.56±64.90 vs. 244.58±85.10, t = -2.024, P = 0.044 ] in the death group was significantly lower than that of the survival group, while the length of ICU stay ( days: 14.33±10.81 vs. 9.11±7.37, t = 2.600, P = 0.010 ), the usage rates of CRRT [ 28.57% ( 10/35 ) vs. 15.20% ( 26/171 ), χ2 = 3.968, P = 0.046 ], vasoactive agents [ 28.57% ( 10/35 ) vs. 12.28% ( 21/171 ), χ2 = 6.511, P = 0.011 ], and blood transfusion ratio [ 42.86% ( 15/35 ) vs. 23.39% ( 40/171 ), χ2 = 7.042, P = 0.008 ] were all obviously higher in the death group than those in the survival group. There were no statistically significant differences in gender, number of operation, the levels of CRP, L/M ratio, lactate and SCr between the two groups ( all P > 0.05 ). Multivariate logistic regression analysis showed that age [ odds ratio ( OR ) = 0.938, 95% confidence interval ( 95%CI ) = 0.898-0.980, P = 0.004 ], APACHE Ⅱ score ( OR = 0.914, 95%CI = 0.839-0.996, P = 0.041 ), the number of dysfunction organs ≥ 3 ( OR = 0.223, 95%CI = 0.066-0.754, P = 0.016 ), and the level of PaO2/FiO2 ( OR = 0.990, 95%CI = 0.982-0.998, P = 0.015 ) were independent risk factors for mortality. Conclusions The age, APACHE Ⅱ score, number of dysfunction organs ≥ 3 and the level of PaO2/FiO2 are of significance in predicting the prognosis of patients with Yangming Fushi syndrome combined with ALI/ARDS. Patients with risk factors of high mortality should be more carefully monitored and treated aggressively.

Objective:To explore the clinical value of stereotactic wire-localized biopsy(SWLB)in digital mammography in the detection of microcalcification of the breast.Methods:A total of 45 patients with nonpalpable breast lesions which were positive for microcalcification by mammography but could not be detected clinically underwent SWLB.Their mammography fndings were analyzed in detail with pathology.Results:Among the 45 cases,13 cases(28.9%)had malignant lesions including ductal carcinoma in situ in 3 cases (20.1%),ductal carcinoma in situ with microinvasion in 4 cases(30.8%),invasive ductal carcinoma in 5 cases (38.5%)and intraductal papillary carcinoma in 1 case(7.7%).Thirty-two cases(71.1%)had benign lesions,2 cases(6.3%)of which were severe atypical hyperplasia.Conclusion:SWLB can accurately guide the surgical excision of nonpalpable breast microcalcification lesions and diagnose microcalcifications exactly,which is helpful for increasing the detection rate of eady-stage breast cancer.