Objective:To establish a method for the separation and determination of ketoprofen enantiomer .Methods:A pre-col-umn derivation RP-HPLC method was used with L-alanine-β-naphthylamine ( L-Ala-β-NA) as the derivation reagent .The RP-HPLC conditions were as follows: a Hypersil ODS-2 column (250 mm ×4.6 mm,5 μm) was applied, the mobile phase was acetonitrile-0.025 mol· L-1 phosphate buffer solution (40∶60, v/v) and the flow rate was 1.0 ml· min-1 , the detection wavelength was set at 245 nm and the column temperature was 25℃.The injection volume was 10μl.Results:Base line separation was achieved for the sep-aration of enantiomer from ketoprofen , and the retention time for S-(+) -ketoprofen and the R-(-) -ketoprofen was 24.2 min and 26.0 min, respectively.Dexketoprofen within the range of 0.025-0.125 mg had a good linear relationship (r=0.998 1) and the aver-age recovery was 90.93%(RSD =4.10%, n=9 ).Conclusion:The method is simple, accurate and reliable, which can be applied in the separation and determination of ketoprofen .

OBJECTIVE:To establish UPLC fingerprint of Gehua formula granules. METHODS:UPLC method were adopted. The determination was performed on Zorbax Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-water at the flow rate of 0.5 mL/min. The detection wavelength was set at 264 nm,column temperature was 25 ℃,and the sample size was 1 μL. Using tectorigenin as reference substance,UPLC chromatograms of 10 batches of Gehua formula granules were determined. The common peak identification and similarity evaluation were conducted by TCM Chromatogram Fingerprint Similarity Evaluation Sys-tem(2004 A edition). RESULTS:14 common peaks were identified in UPLC chromatograms of 10 batches of Gehua formula gran-ules and similarities were all higher than 0.90. UPLC chromatograms of 10 batches of samples were in good agreement with control fingerprint. CONCLUSIONS:Established UPLC fingerprint can provide reference for identification and quality evaluation of Gehua formula granules.

Objective To evaluate the effects of sevoflurane postconditioning on the oncosis and apoptosis in cardiomyocytes during ischemia-reperfusion (I/R) in isolated rat hearts and the role of extracellular signalregulated protein kinase 1/2 (ERK1/2) signal transduction pathway in it.Methods Seventy-two isolated rat hearts perfused in a Langendorff apparatus were randomly divided into 6 groups (n =12 each) using a random number table:sham operation group (group S),myocardial I/R group (group I/R),sevoflurane postconditioning group (group SP),PD98059 vehicle dimethyl sulfoxide (DMSO) group (group DMSO),selective ERK1/2 inhibitor PD98059 group (group PD),and sevoflurane postconditioning + PD98059 group (group SP + PD).The hearts were subjected to ischemia for 30 min followed by 2 h reperfusion in the other groups except group S.In SP,DMSO and PD groups,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane,DMSO (＜0.2％) and PD98059 (20 μmol/L),respectively,for 15 min starting from the end of ischemia until 15 min of reperfusion,and then with plain K-H solution for 105 min.In group SP+ PD,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane and PD98059 for 15 min starting from the end of ischemia until 15 min of reperfusion.Myocardial infarct size and expression of porimin and caspase-8 proteins (by Western blot) were measured at the end of reperfusion.Results Compared with S group,the myocardial infarct size was significantly increased,and the expression of porimin and caspase-8 proteins was up-regulated in the other groups (P ＜ 0.05).Compared with I/R group,the myocardial infarct size was significantly decreased,and the expression of porimin and caspase-8 proteins was down-regulated in group SP (P ＜ 0.05),and no significant changes were found in the other groups (P ＞ 0.05).Conclusion Sevoflurane postconditioning can activate ERK1/2 signal transduction pathway and inhibit the oncosis and apoptosis in cardiomyocytes,thus attenuating I/R injury in isolated rat hearts.

AIM To establish the HPLC fingerprints of Cajanus cajan (L.) Millsp.leaves and to determine the contents of orientoside and luteolin.METHODS The analysis of 65％ methanol extract from C.cajan leaves was performed on a 25 ℃ thermostatic Agilent Zorbax SB-C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-1％ acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS There were twenty-one common peaks in ten batches of samples (S1-S10),whose similarities were more than 0.950,except for that of S3 (0.516).Orientoside and luteolin showed good linear relationships within the ranges of 0.089 5-3.960 μg and 0.015 5-0.408 μg,whose average recoveries were 99.43％ (RSD =1.32％) and 98.50％ (RSD =0.82％),respectively.The contents of two constituents in the samples from three growing areas (Guangdong,Yunnan and Hainan) showed obvious differences.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of C.cajan leaves.

OBJECTIVETo clone differentially expressed cDNA sequences involved in malignant transformation induced by benzo(a)pyrene metabolite dihydroxyepoxy benzo pyrene (BPDE).

METHODThe malignant transformation of human bronchial epithelial cell line 16HBE induced by BPDE in vitro was used as a model for comparing gene expression between the transformed cells and controls. cDNA representational difference analysis (cDNA-RDA) was performed to isolate differentially expressed cDNA fragment in transformed cells. The cDNA fragments were ligated to pGEM-T vector and transformed into JM109 bacteria. The plasmid DNA were sequenced and compared with data in GenBank by BLASTN.

RESULTSFive cDNA sequences were found to be novel ones and were registered in dbest database, which assigned accession numbers in GenBank are BG354691, BG354692, BG354693, BG354694 and BG354695, respectively. Eight of the remaining cDNA sequences showed sequence homology to those previously reported such as ribosomal protein S23, MLN137, ACTN4, transforming growth factor and G protein gene.

CONCLUSIONSThese 13 genes may be involved in BPDE-induced malignant transformation, but their biological characteristics and functions are left to further studies.

Benzopyrenes ; metabolism ; pharmacology ; Carcinogens ; pharmacology ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; Cells, Cultured ; Cloning, Molecular ; DNA, Complementary ; analysis ; drug effects ; DNA, Neoplasm ; analysis ; Gene Expression ; drug effects ; Humans

Aim To study the effect of melatonin on expression and activity of myosin light chain kinase in the artery wall of atherosclerotic rabbits.Methods The rabbit model of atherosclerosis was induced by a high-cholesterol diet.The blood lipid levels were assayed in the serum of each group.MLCK expression was detected by Western blot and immunohistochemical method.MLCK activity was measured by ?-32P-ATP incorporation into myosin light chain.Results The atherosclerosis model was established successfully.The levels of lipids decreased after MLT treatment.After fed with cholesterol for twelve weeks,the expression and activity of MLCK in the artery of atherosclerotic rabbits increased markedly,whereas there was no obvious difference in expression of MLCK in the artery of atherosclerotic rabbits fed with cholesterol and melatonin for twelve weeks compared with that of control.Conclusions It was suggest that high expression and activity of MLCK in the artery might be closely correlated with the development of atherosclerosis.Melatonin played an important role in inhibiting the development of atherosclerosis by decreasing the expression and activity of MLCK.

OBJECTIVETo study the gene mutation and streptomycin, isoniazid or rifampicin resistance of Mycobacterium isolated from silico-tuberculosis patient's sputum so as to find a more effective therapy for this disease.

METHODSMycobacteria tuberculosis were separated from 96 coal worker with silico-tuberculosis firstly. Then rpsL, KatG and rpoB fragments of genome were copied with PCR and compared their SSCP profiles with standard strains.

RESULTS67 strains of streptomycin, isoniazid or rifampicin resistant Mycobacteria tuberculosis were found in routine drug resistance test, with the percentages of 80.5% (54/67), 58.2% (39/67) respectively. PCR-SSCP showed that out of 67 drug-resistant strains, 66(98.5%) of rpsL, 47(70.1%) of rpoB and 42(62.7%) of KatG appeared abnormal.

CONCLUSIONMost of the resistant strains appeared gene mutation. The mution rates were higher than the results from routine drug resistance test.

Coal ; Drug Resistance, Bacterial ; Humans ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Silicotuberculosis ; microbiology ; Sputum ; microbiology

Objective:To analyze the risk factors related to the prognosis of patients with sepsis in intensive care unit (ICU), construct a nomogram model, and verify its predictive efficacy.Methods:A retrospective cohort study was conducted using data from Medical Information Mart for Intensive Care-Ⅳ 0.4 [MIMIC-Ⅳ (version 2.0)]. The information of 6 500 patients with sepsis who meet the diagnostic criteria of Sepsis-3 were collected, including demography characteristics, complications, laboratory indicators within 24 hours after ICU admission, and final outcome. Using a simple random sampling method, the patients were divided into a training set and a validation set at a ratio of 7∶3. The restricted cubic spline (RCS) was used to explore whether there was a linear relationship between each variable and the prognosis, and the nonlinear variables were truncated into categorical variables. All variables were screened by LASSO regression and included in multivariate Cox regression analysis to analyze the death risk factors in ICU patients with sepsis, and construct a nomograph. The consistency index, calibration curve and receiver operator characteristic curve (ROC curve) were used to evaluate the prediction efficiency of nomogram model. The decision curve analysis (DCA) was used to validate the clinical value of the model and its impact on actual decision-making.Results:Among 6 500 patients with sepsis, 4 551 were in the training set and 1 949 were in the validation set. The 28-day, 90-day and 1-year mortality in the training set were 27.73% (1?262/4?551), 34.76% (1?582/4?551), and 42.98% (1?956/4?551), respectively, those in the validation set were 27.24% (531/1?949), 33.91% (661/1?949), and 42.23% (823/1?949), respectively. Both in training set and the validation set, compared with the final survival patients, the death patients were older, and had higher sequential organ failure assessment (SOFA) score and simplified acute physiology scoreⅡ (SAPSⅡ), more comorbidities, less urine output, and more use of vasoactive drugs, kidney replacement therapy, and mechanical ventilation. By RCS analysis, the variables with potential nonlinear correlation with the prognosis risk of septic patients were transformed into categorical variable. The variables screened by LASSO regression were enrolled in the multivariate Cox regression model. The results showed that age [hazard ratio ( HR) = 1.021, 95% confidence interval (95% CI) was 1.018-1.024], SOFA score ( HR = 1.020, 95% CI was 1.000-1.040), SAPSⅡ score > 44 ( HR = 1.480, 95% CI was 1.340-1.634), mean arterial pressure (MAP) ≤ 75 mmHg (1 mmHg ≈ 0.133 kPa; HR = 1.120, 95% CI was 1.026-1.222), respiratory rate (RR; HR = 1.044, 95% CI was 1.034-1.055), cerebrovascular disease ( HR = 1.620, 95% CI was 1.443-1.818), malignant tumor ( HR = 1.604, 95% CI was 1.447-1.778), severe liver disease ( HR = 1.330, 95% CI was 1.157-1.530), use of vasoactive drugs within 24 hours ( HR = 1.213, 95% CI was 1.101-1.336), arterial partial pressure of oxygen (PaO 2; HR = 0.999, 95% CI was 0.998-1.000), blood lactic acid (Lac; HR = 1.066, 95% CI was 1.053-1.079), blood urea nitrogen (BUN) > 8.9 mmol/L ( HR = 1.257, 95% CI was 1.144-1.381), total bilirubin (TBil; HR = 1.023, 95% CI was 1.015-1.031), and prothrombin time (PT) > 14.5 s ( HR = 1.232, 95% CI was 1.127-1.347) were associated with the death of ICU patients with sepsis (all P < 0.05). Based on the above factors, a nomogram model was constructed, and the model validation results showed that the consistency index was 0.730. The calibration curve showed a good consistency between the predicted results of the nomogram model and observed results in the training and validation sets. ROC curve analysis showed that the area under the ROC curve (AUC) predicted by the nomogram model in the training set and the validation set for 28-day, 90-day and 1-year death risk was 0.771 (95% CI was 0.756-0.786) and 0.761 (95% CI was 0.738-0.784), 0.777 (95% CI was 0.763-0.791) and 0.765 (95% CI was 0.744-0.787), 0.677 (95% CI was 0.648-0.707) and 0.685 (95% CI was 0.641-0.728), respectively. DCA analysis showed that the nomogram model had significant net benefits in predicting 28-day, 90-day, and 1-year death risk, verifying the clinical value of the model and its good impact on actual decision-making. Conclusions:The death risk factors related to ICU patients with sepsis include age, SOFA score, SAPSⅡ score > 44, MAP ≤ 75 mmHg, RR, cerebrovascular disease, malignant tumors, severe liver disease, use of vasoactive drugs within 24 hours, PaO 2, Lac, BUN, TBil, PT > 14.5 s. The nomogram model constructed based on this can predict the death risk of ICU patients with sepsis.

Objective To explore the disease cognition and demand of health education in outpatients with foodborne diarrhea so as to provide a basis for prevention and health education of foodborne diarrhea. Methods A total of 342 patients with foodborne diarrhea were selected by convenience sampling from 1st April 2016 to 1st October 2016. They were investigated with the self-designed disease cognition scale for patients with foodborne diarrhea and the demand of health education scale. Results When foodborne diarrhea occurred, treatment behavior patients preferred was without treatment (41.81%). Before the hospital visit,most of them used antidiarrheal agent (47.37%) and antibiotics (31.87%). During diarrhea,fasting was in the majority (69.88%). The influencing factors of knowledge condition in patients with foodborne diarrhea were ages, degrees of education,frequency of diarrhea in the last six months (P< 0.01). The high demand of patients with foodborne diarrhea on knowledge was the prevention of foodborne diarrhea (89.47%),treatment methods of diarrhea (88.30%),diet management (82.75%). The low demand on knowledge was the cause (35.96%) and clinical manifestation (31.58%) of foodborne diarrhea. Conclusions The disease cognition of outpatients with foodborne diarrhea is low. They are with high demand on prevention and treatment of foodborne diarrhea. Medical staff should provide targeted health education according to ages,degrees of education,actual demand and so as to prevent foodborne diarrhea.

Objective To investigate the ADC value and intravoxel incoherent motion (IVIM) parameters to predict the early response to chemotherapy in patients with locally advanced non-small cell lung cancer (NSCLC). Methods Twenty-six patients with pathologically confirmed NSCLC in our hospital from June 2015 to January 2017 were prospectively studied. MR routine scan, DWI and IVIM scans were performed before and at the end of the first cycle of chemotherapy. The ADC value, pure diffusion parameter (D), perfusion parameter (D*) and perfusion fraction (f) before and after chemotherapy were recorded respectively, and the maximum diameter of tumor (MDMRI) was measured on T2WI. The effective group and the ineffective group were divided according to RECIST 1.1. Intra- and interobserver reproducibility were assessed with intraclass correlation coefficient (ICC). The differences of the parameters and their change rate were compared pre-and posttreatment, in the effective and the ineffective groups by using t test. Receiver operating characteristic (ROC) curves were generated to explore the optimal parameters and thresholds for early prediction of the efficacy of chemotherapy. Z test was used to compare the differences in the areas under the ROC curves. Results The repeatability of D*was relatively poor (ICC 0.507-0.716). The ADC and D values of the effective group before chemotherapy were (1.28±0.21)×10-3 and (1.05±0.14)×10-3mm2/s, respectively, and those of the ineffective group were (1.55 ± 0.25) × 10-3 and (1.29 ± 0.18) × 10-3mm2/s, respectively. The differences between two groups were statistically significant (t=-2.989, -3.755, respectively, P all<0.05). While there was no significant difference in f between the effective and ineffective groups before chemotherapy (t=-1.034, P=0.312). The ADC, D and f values of the effective group before chemotherapy were lower than those after chemotherapy, and the differences were statistically significant (t=-3.969,-5.617 and-2.876, respectively, P all<0.05). While there was no significant difference in ADC and D values in ineffective group (t=-0.737, -0.866, P>0.05, respectively). The difference of f was statistically significant (t=-2.731, P<0.05).ΔADC％,ΔD％andΔMDMRI％in effective group were (23.90 ± 22.28)％, (32.58 ± 20.63)％ and (18.77 ± 10.36)％, respectively, and those in ineffective group were (1.19 ± 11.35)％, (6.22 ± 21.63)％ and (9.35 ± 8.26)％, respectively. The differences between two groups were statistically significant (t=3.393, 3.154 and 2.487, respectively, P all<0.05). There was no significant difference inΔf％between two groups (t=1.346, P=0.191). ROC curve analysis showed that the area under curve (AUC) of preADC, preD, ΔADC％, ΔD％ and ΔMDMRI％ was 0.788, 0.848, 0.800, 0.830 and 0.727, respectively. When setting the preD=1.20×10-3 mm2/s as the boundary under the largest AUC, however, there was no significant differences in AUC compared with preADC, ΔADC％, ΔD％ and ΔMDMRI％ (Z=1.336, 0.363, 0.136, 1.395, P>0.05). Conclusion The ADC value, D value before chemotherapy and the early change rate after chemotherapy can better predict the early response of chemotherapy in NSCLC patients.