Genomic DNA of the two bacterial strains including Escherichia coli DH5? and Enterobacter cloacae E 26R were amplified by PCR with specific primers of ERIC sequence Each strain showed the stable and unique DNA fingerprint when PCR products were analyzed in agarose gel electrophoresis The stability of the genomic DNA fingerprint wasn't influenced by the templet from 10ng to 100ng In the fingerprints there were characteristic bands The unique band was influenced difficultly due to the change of the environment and condition of experiment DH5? and E 26R were mixed proportionately, and examined by ERIC PCR The result showed that the DNA fingerprints of the mixture were the superposition of each pure bacterium's Analysis of the main bands showed that DH5? can be examined by ERIC PCR when its concentration gets to 0 5% of the total mixed bacteria The study provides the reference for the research of soil microbe and environmental microbe, and especially can be used for quick identification and examination of the sundry bacterial contamination in the fermentation industry

Objective To compare temperature curve and ablation zone between 915 MHz and 2450 MHz cooled shaft microwave antenna in ex vivo porcine livers.Methods The 915 MHz and 2450 MHz microwave ablation and thermal monitor system were used in this study.A total of 56 ablation zones and 280 temperature data were obtained in ex vivo porcine livers.The output powers were 50,60,70,and 80W and the setting time was 600s.The temperature curve of every temperature spot,the short- and long-axis diameters of the coagulation zones were recorded and measured.Results At all four power output settings,the peak temperatures of every temperature spot had a tendency to increase accordingly as the output power was increased,and except for 5 mm away from the antenna,the peak temperatures for the 915 MHz cooledshaft antenna were significantly higher than those for the 2450 MHz cooled-shaft antenna (P ＜0.05).Meanwhile,the short- and long-axis diameters for the 915 MHz cooled-shaft antenna were significantly larger than those for the 2450 MHz cooled-shaft antenna ( P ＜0.05).Conclusions The 915 MHz cooledshaft antenna can yield a significantly larger ablation zone and achieve higher temperature in ablation zone than 2450 MHz cooled-shaft antenna in ex vivo porcine livers.

OBJECTIVEThis paper is aimed to explore the adverse reaction condition of Shuanghuangli an injection with three common used signal detecting methods based on SRS database of Jiangsu province, and to evaluate the performance of three methods.

METHODThree methods would be used to detect the signals based on the SRS database of Jiangsu province. Consistency of the results of these three methods with that proved in descriptions was evaluated by Kappa test. The trend graph of the confidence intervals of several time points was used to demonstrate the trend of the signal.

RESULTThe PRR method was consistent with ROR method in high degree in any situation. The results of BCPNN method was close to PRR and ROR method only when the related report count was larger. PRR and ROR methods had higher false positive rate than BCPNN method.

CONCLUSIONPRR or ROR method is proposed for signal detecting when the report count is large. BCPNN method is proposed for trend demonstration of signal with graph.

Objective: PS128 is a novel psycho biotic strain, it has been reported to play an important role in neuropsychiatric disorders. This study investigated the clinical effect of PS128 supplementation on patients with anxiety. Methods: A total of 200 patients with anxiety were recruited, and divided into two groups (n = 100/group). The control group received oral treatment with citalopram, and the PS128 group received PS128 capsules based on citalopram treatment. Hamilton Anxiety Scale (HAMA) and Self-Rating Anxiety Scale (SAS) were used to evaluate the anxiety levels.After 2 months of continuous administration, clinical efficacy was evaluated according to HAMA score. Results: There was no significant difference in HAMA and SAS scores between the two groups before treatment. With the treatment prolonged, the HAMA and SAS score decreased gradually in both control and PS128 groups, and the decrease rate of PS128 group was significantly greater than that of the control group. The clinical effective rates of PS128 group were higher than those in the control group, high levels of clinical cure rate were also detected in the PS128 group. Compared with the control group (22%), the incidence of adverse reactions was significantly reduced for patients in the PS128 group (4%). Conclusion The treatment effect of citalopram combined with PS128 against anxiety is satisfactory clinically. It can greatly improve the anxiety symptoms of patients, increase the cure rate, reduce adverse reactions.

In post-marketing study of traditional Chinese medicine (TCM), pharmacoeconomic evaluation has an important applied significance. However, the economic literatures of TCM have been unable to fully and accurately reflect the unique overall outcomes of treatment with TCM. For the special nature of TCM itself, we recommend that Markov model could be introduced into post-marketing pharmacoeconomic evaluation of TCM, and also explore the feasibility of model application. Markov model can extrapolate the study time horizon, suit with effectiveness indicators of TCM, and provide measurable comprehensive outcome. In addition, Markov model can promote the development of TCM quality of life scale and the methodology of post-marketing pharmacoeconomic evaluation.
Economics, Pharmaceutical ; Markov Chains ; Medicine, Chinese Traditional ; economics ; Product Surveillance, Postmarketing

Objective To investigate the biological characteristics of monoclonal antibodies against human liver cancer stem cells and its therapeutic effect in combination with cisplatin in the treatment of hepatocellular carcinoma. Methods Cell culture in serum?free medium and PKH26 staining were used to determine the existence of cancer stem cells in human liver Bel7402?V3 cell line. The co?expression of antigen recognized by monoclonal antibody ( McAb ) 15D2 and epithelial specific antigen ( ESA ) and PKH26?positive cells in the Bel7402?V3 cells were detected by immunofluorescence assay. Serum?free suspension culture was used to detect the self?renewal ability of 15D2?positive Bel7402?V3 cells sorted by flow cytometry and the effect of 15D2 on the self?renewal ability of Bel7402?V3 cells. The effect of 15D2 on cisplatin resistance in the cells was examined by CCK8 method. The inhibitory effect of 15D2 combined with cisplatin on the transplanted tumor growth in mice was also observed. Results Single PKH26?positive cells were observed in the Bel7402?V3 cell spheroids cultured for 11 days. Immunofluorescence assay showed that the 15D2?recognized antigen could be conjugated with PKH26 and ESA and co?localized on Bel7402?V3 cells. The spheroid formation rate of 15D2?positive cells in serum?free medium was significantly higher than that of 15D2?negative cells [(30.4±3.4)% vs. (8.8±1.8)%,P<0.01]. The cisplatin resistance of 15D2?positive cells was obviously higher than that of 15D2?negative cells (IC50:1.014μmol/L vs. 0.365μmol/L). McAb 15D2 significantly suppressed the spheroid formation of Bel7402?V3 cells, with an inhibition rate of 37.5%. McAb 15D2 also notably inhibited the cisplatin resistance of Bel7302?V3 cells. The IC50 was 0.211μg/ml in the 15D2 group and 0. 325 μg/ml in the control group. The mouse experiment showed that the tumor growth rates of 50 mg/kg, 25 mg/kg and 12.5 mg/kg 15D2?treatment groups were 82.6%, 71.4% and 60.0%, respectively; that of the 50 mg/kg 15D2 + cisplatin group was 91. 0%, and that of the cisplatin monotherapy was 56. 7%. Conclusion McAb 15D2 is a functional monoclonal antibody targeting liver cancer stem cells, which could be a potential monoclonal antibody drug for the stem cell?targeted therapy of liver cancer.

Objective To investigate the biological characteristics of monoclonal antibodies against human liver cancer stem cells and its therapeutic effect in combination with cisplatin in the treatment of hepatocellular carcinoma. Methods Cell culture in serum?free medium and PKH26 staining were used to determine the existence of cancer stem cells in human liver Bel7402?V3 cell line. The co?expression of antigen recognized by monoclonal antibody ( McAb ) 15D2 and epithelial specific antigen ( ESA ) and PKH26?positive cells in the Bel7402?V3 cells were detected by immunofluorescence assay. Serum?free suspension culture was used to detect the self?renewal ability of 15D2?positive Bel7402?V3 cells sorted by flow cytometry and the effect of 15D2 on the self?renewal ability of Bel7402?V3 cells. The effect of 15D2 on cisplatin resistance in the cells was examined by CCK8 method. The inhibitory effect of 15D2 combined with cisplatin on the transplanted tumor growth in mice was also observed. Results Single PKH26?positive cells were observed in the Bel7402?V3 cell spheroids cultured for 11 days. Immunofluorescence assay showed that the 15D2?recognized antigen could be conjugated with PKH26 and ESA and co?localized on Bel7402?V3 cells. The spheroid formation rate of 15D2?positive cells in serum?free medium was significantly higher than that of 15D2?negative cells [(30.4±3.4)% vs. (8.8±1.8)%,P<0.01]. The cisplatin resistance of 15D2?positive cells was obviously higher than that of 15D2?negative cells (IC50:1.014μmol/L vs. 0.365μmol/L). McAb 15D2 significantly suppressed the spheroid formation of Bel7402?V3 cells, with an inhibition rate of 37.5%. McAb 15D2 also notably inhibited the cisplatin resistance of Bel7302?V3 cells. The IC50 was 0.211μg/ml in the 15D2 group and 0. 325 μg/ml in the control group. The mouse experiment showed that the tumor growth rates of 50 mg/kg, 25 mg/kg and 12.5 mg/kg 15D2?treatment groups were 82.6%, 71.4% and 60.0%, respectively; that of the 50 mg/kg 15D2 + cisplatin group was 91. 0%, and that of the cisplatin monotherapy was 56. 7%. Conclusion McAb 15D2 is a functional monoclonal antibody targeting liver cancer stem cells, which could be a potential monoclonal antibody drug for the stem cell?targeted therapy of liver cancer.

Objective:To explore the expression of fructose bisphosphate aldolase A (ALDOA) in the bone marrow of patients with acute myeloid leukemia (AML) and the correlation with clinical features and prognosis.Methods:The bone marrow samples of 90 newly diagnosed AML (non-acute promyelocytic leukemia) patients and 18 allogeneic hematopoietic stem cell transplantation donors who were treated from January 2013 to December 2015 in the First Affiliated Hospital of Zhengzhou University and the Children's Hospital Affiliated to Zhengzhou University were collected. The relative expression level of ALDOA mRNA in bone marrow samples was detected by using real-time quantitative polymerase chain reaction (qRT-PCR). Clinical data of these patients were retrospectively analyzed, and the patients were divided into continuous complete remission (CR) group and refractory recurrent (RR) group according to the clinical response and follow-up results. The differences of the relative expression level of ALDOA mRNA between AML group and the normal control group, CR group and RR group were analyzed. Univariate and multivariate Cox regression risk model were used for analysis of factors influencing prognosis of AML patients.Results:The relative expression level of ALDOA mRNA in AML group was higher than that in normal control group [(5.71±0.44) vs. (1.10±0.08), t = 4.74, P<0.001]. The relative expression level of ALDOA mRNA in the RR group was higher than that in the CR group [(6.69±0.67) vs. (4.30±0.36) , t = 2.79, P < 0.001]. In addition, there were statistically significant differences in the proportion of patients with ALDOA mRNA high expression and those with ALDOA mRNA low expression stratified by the number of white blood cell, the proportion of bone marrow blasts and whether complete remission could be achieved or not after 1 course of induction therapy (all P < 0.05). Overall survival in patients with ALDOA high expression was worse than that in patients with ALDOA low expression ( χ2 = 5.59, P = 0.018). Multivariate analysis showed that white blood cell count, prognosis stratification, whether complete remission could be achieved or not after 1 course of induction therapy and ALDOA expression were the independent prognostic factors for the death of AML patients (all P < 0.05). Conclusions:ALDOA may play an important role in the development and progression of AML, and the expression level of ALDOA in the bone marrow can be used as an index for the prognosis assessment of AML patients and may be a potential therapeutic target for AML.

OBJECTIVE To establish two-dimensional liquid chromatography method for the determination of imipenem blood concentration and apply it in clinical practice. METHODS The method for the determination of imipenem blood concentration was established based on automatic two-dimensional liquid chromatography. The targets were extracted by 1-dimensional column Aston SNCB （50 mm ×4.6 mm， 5 μm） and further separated and determined by 2-dimensional column Aston SCB （250 mm×4.6 mm， 5 μm）. The 1-dimensional mobile phase was imipenem-1D mobile phase [acetonitrile-methanol-water （15∶10∶75， V/V/V）] with a flow rate of 1.0 mL/min； 2-dimensional mobile phase was 72%OPI-1 organic mobile phase （chromatographic grade methanol）-20% BPI-1 alkaline mobile phase [water （containing 20.0 mmol/L ammonium phosphate， pH adjusted to 7.2 with triethylamine）]-8%API-1 acidic mobile phase [water （containing 20.0 mmol/L ammonium phosphate， pH adjusted to 3.0 with phosphoric acid）] with a flow rate of 1.0 mL/min； the column temperature was 40 ℃， UV detection wavelength was 310 nm and injection volume was 100 μL. Elution procedure： 1-dimensional column consisted of imipenem-1D mobile phase with eluting for 0-3.40 min； 2-dimensional column consisted of 72% OPI-1 organic mobile phase-20%BPI-1 alkaline mobile phase-8%API-1 acidic mobile phase with eluting for 3.40-11.00 min. RESULTS The linear range of imipenem was 0.171-18.570 μg/mL （R 2＝0.999 9） with the lower limit of quantification for 0.171 μg/mL； the recovery rate ranged from 93.47% to 106.16%（ n＝5） and the RSDs of both intra-day and inter- day precision were below 15% （n＝5）. The minimum concentration of imipenem in 51 patients ranged from 0 to 19.57 μg/mL. CONCLUSIONS The established method is simple and fast with the large scale of sample， and can be used for the imipenem blood concentration monitoring in patients with severe infection.

OBJECTIVE： To investigate the rationality of TLC identification method （3） of （R，S）-epigoitrin in Isatis indigotica stated in 2015 edition of Chinese Pharmacopeia （partⅠ） （later abbreviated as pharmacopeia）， and make some improvements. METHODS： Three batches I. indigotica were collected and prepared into decoction pieces according to the processing method of I. indigotica in pharmacopoeia. TLC identification of （R，S）-goitrin in I. indigotica decoction piece and medicinal material were conducted according to identification method （3） in pharmacopeia （80％ ethanol as solvent for sample treatment， ultrasound extraction）； the rationality of pharmacopoeia method was investigated. Then the method was improved by changing the extraction solvent and pretreatment method （method one： using water as solvent， ultrasound extraction； method two： soaking in water for 1 h， then adding into methanol， ultrasound extraction； method three： the sample was wetted and then dried， using 80％ methanol as solvent， ultrasound extraction） of samples， and the optimal method was verified. According to the optimal method， the TLC identification of （R，S）-goitrin was detected by using chromatographic plates from different manufacturers， under the conditions of low temperature and low humidity （7 ℃， relative humidity 48％） and high temperature and high humidity （35 ℃， relative humidity 75％） respectively，to investigate the durability of the method. RESULTS： According to the method of pharmacopeia， in the chromatograms of decoction pieces， the same color spots appeared at the corresponding chromatographic positions of reference substance， but no corresponding spots appeared in the medicinal material chromatograms. After the samples were treated by three improvement methods， in medicinal material chromatograms， the same color spots appeared in the corresponding chromatographic positions of reference substances. There were single chromatographic spot after medicinal materials were treated with method one， and there were more spots after medicinal materials were treated with method two and three， and method two consumed less time than method three. The results of validation tests and method durability tests  showed that after the treatment of I. indigotica and its decoction pieces according to method two， the same color spots appeared in the corresponding positions of the decoction pieces and the medicinal materials chromatograms as those of the control. CONCLUSIONS： The improved TLC identification method is effective， the chromatographic spots are clear， and the repeatability is good.