Development and comparative study of determination of Escherichia coli in different processed products of Angelicae Sinensis Radix based on fluorescence quantitative PCR / 中草药

Objective: To make quantitative analysis for the quantity of Escherichia coli in Angelicae Sinensis Radix (ASR) and its processed products. Methods: The fluorescence quantitative PCR method was established to quantitatively analyze E. coli in ASR from different processed products, different producing areas, different enterprises and different storage time. Results: The number of E. coli in different processed products was ranked as follows: ASR > ASR stir-frying with soil > ASR stir-frying with wine. And the number of E. coli in the three producing areas of ASR in Min County of Gansu Province was less than that in other producing areas. Compared with the retail enterprises, the number of E. coli in ASR and ASR stir-frying with wine was less in production and sale enterprises. Different storage time had certain effect on the number of E. coli in ASR and ASR stir-frying with wine. With the increase of storage time, the number of E. coli also increased. Plate counting method and fluorescence quantitative PCR method were carried out at the same time for some representative samples. The results showed that the results of the plate counting method were mostly negative, and the results of the fluorescence quantitative PCR were positive. Conclusion: The quantitative fluorescence PCR method established in this paper is superior to the plate counting method in specificity, sensitivity, reliability, and reporting cycle, which can provide an effective method for rapid and accurate quantitative detection of E.coli in different processed products of ASR.

Development and Comparison of Determination of Staphylococcus aureus in Different Processed Products of Angelicae Sinensis Radix Based on Real-time PCR Technique / 中国实验方剂学杂志

Objective:To quantitatively analyze the changes of Staphylococcus aureus in different processed products of Angelicae Sinensis Radix. Method:The real-time fluorescence quantitative polymerase chain reaction method （Real-time PCR） was established to quantitatively analyze S. aureus in Angelicae Sinensis Radix decoction pieces which bought from different producing areas， different enterprises and different storage time. The fluorescence quantitative reaction system was SYBR Premix Ex Taq Ⅱ of 10 μL， each of forward primer and reverse primer （10 μmol·L-1） of 0.8 μL， template/genome DNA of 1 μL， double distilled water of 7.4 μL. The reaction conditions of the fluorescence quantitative amplification curve were pre-denaturing for 30 s at 94 ℃， denaturing for 10 s at 94 ℃， annealing for 12 s at 60 ℃， extensing for 30 s at 72 ℃， cycling 45 times， single-point detection signal at 72 ℃. The melting curve was made from 72 ℃， and the step temperature of 0.5 ℃ was kept for 15 s to collect fluorescence. According to the results of Real-time PCR， representative samples were selected from Angelicae Sinensis Radix decoction pieces for comparison between plate counting method and Real-time PCR. Result:The content of S. aureus in different processed products was sorted by rank of raw Angelicae Sinensis Radix>soil-fried Angelicae Sinensis Radix>wine-processed Angelicae Sinensis Radix. The content of S. aureus was the lowest in the samples from Weiyuan area of Gansu province by comparing with other producing areas. Compared with the retail enterprises， the content of S. aureus in raw products and wine-processed products from production and sale enterprises was lower. Different storage time had certain effect on the content of S. aureus in raw products and wine-processed products， and the content of S. aureus increased with the increase of storage time. The detection results of plate counting method were 3-4 orders of magnitude lower than that of Real-time PCR. Conclusion:The established Real-time PCR is superior to plate counting method in specificity， sensitivity， reliability and reporting period， which can provide an effective method for rapid and accurate quantitative detection of S. aureus in different processed products of Angelicae Sinensis Radix.

UPLC fingerprint and multi-components content determination of different processed products of Angelica sinensis / 中国中药杂志

Ten batches of Angelica sinensis from three producing areas( Tuoxiang,Minxian and Weiyuan of Gansu province) were selected as the research objects,and processed into raw A. sinensis,A. sinensis with alcohol,and A. sinensis with soil respectively through the standard processing methods. Ultra-high performance liquid chromatography( UPLC) was used to establish fingerprint for three processed products of A. sinensis,and determine the contents of 9 phenolic acids and phthalide compounds. The similarity was analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine,which showed that the chromatographic peaks of the same processed samples of A. sinensis were basically similar,with all similarities greater than 0. 950. The difference between different processed products and their control spectra was not obvious,with all similarities also higher than 0. 950.On the basis of using principal component analysis( PCA) and OPLS-DA to seek the difference components between groups,the improved distance coefficient method can be used to effectively distinguish the three processed products of A. sinensis by fingerprint similarity. At the same time,the determination method of nine phenolic acids and phthalide in A. sinensis was established by UPLC,and the comparison between different processed products was carried out. The results showed that the content of various components was changed as compared with the raw A. sinensis. The contents of coniferyl ferulate and ligustilide in the A. sinensis with alcohol were increased significantly,and the content of coniferyl ferulate was obviously increased in A. sinensis with soil. The method established in this paper can effectively distinguish different processed products of A. sinensis and determine the content of the main components in them.

Efficiency evaluation of China's heath resources allocation under medical expense control constraint / 中国卫生政策研究

Objective:The main objective of this study is to evaluate the efficiency of China's heath resources al-location during 2004 to 2015 under the constraint of medical expenses control. Methods:To evaluate the efficiency of China's heath resources allocation,the undesired output Slacks-Based Measure(SBM) model was used. Results:As per the findings of this study,the efficiency of heath resources allocation at the national and regional levels of the undesired output SBM model was significantly lower than that of the traditional CCR model during 2004 to 2015. The non-expec-ted output redundancy rate and expected output deficiency rate at the national and regional levels were much greater than the input redundancy rate of health resources allocation of the national and regional levels. In addition,with re-dundancy rate introduction,the losses of health resource allocation efficiency in different provinces within the region were not the same. Conclusions:The efficiency of China's heath resources allocation was overestimated by the tradition-al DEA model,which was less sensitive to the change in its characteristics..Giving priority to non-expected output re-dundancy and expected output deficiency are the main reasons for the loss of health resource allocation in china,and are considered as internal and external improvement priorities for the performance of health resource allocation. It was suggested to establish the performance evaluation system of health resource allocation,which includes the integration of health resources input,expected-output and non-expected output,and evaluation techniques;and a mechanism for per-formance assessment and evaluation,supervision and feedback of health resource allocation should be established;and improve the implication of regional medical and health planning policies.

Associations between epidermal growth factor receptor gene mutation and serum tumor markers in advanced lung adenocarcinomas: a retrospective study / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the associations between epidermal growth factor receptor (EGFR) gene mutations and serum tumor markers in advanced lung adenocarcinomas.</p><p><b>METHODS</b>We investigated the association between EGFR gene mutations and clinical features, including serum tumor marker levels, in 97 advanced lung adenocarcinomas patients who did not undergo the treatment of EGFR tyrosine kinase inhibitors. EGFR gene mutation was detected by real-time PCR at exons 18, 19, 20, and 21. Serum tumor marker concentrations were analyzed by chemiluminescence assay kit at the same time.</p><p><b>RESULTS</b>EGFR gene mutations were detected in 42 (43%) advanced lung adenocarcinoma patients. Gender (P=0.003), smoking status (P=0.001), and abnormal serum status of carcinoembryonic antigen (CEA, P=0.028) were significantly associated with EGFR gene mutation incidence. Multivariate analysis showed the abnormal CEA level in serum was independently associated with the incidence of EGFR gene mutation (P=0.046) with an odds ratio of 2.613 (95% CI: 1.018-6.710). However, receiver operating characteristic (ROC) curve analysis revealed CEA was not an ideal predictive marker for EGFR gene mutation status in advanced lung adenocarcinoma (the area under the ROC curve was 0.608, P=0.069).</p><p><b>CONCLUSIONS</b>EGFR gene mutation status is significantly associated with serum CEA status in advanced lung adenocarcinmoas. However, serum CEA is not an ideal predictor for EGFR mutation.</p>

Effect of rapamycin on apoptosis in human myelodysplastic syndrome cell line MUTZ-1 and its possible mechanisms / 中国实验血液学杂志

The aim of this study was to investigate the effect of rapamycin on cell growth and apoptosis in the myelodysplastic syndrome (MDS) cell line MUTZ-1 and possible mechanism. MUTZ-1 cells were treated with rapamycin, cell proliferation capability was determined with MTT, protein expression including Annexin V/PI, caspase 3, PTEN, p-Akt, p-mTOR and the cell cycle were analyzed with flow cytometry. The results indicated that the proliferation of MUTZ-1 cells was inhibited by rapamycin in concentration-and time-dependent manners (r=0.67, 0.61, 0.72). After treatment with rapamycin for 24-72 hours, cell count in G0/G1 were significantly higher than that of the control (p<0.01), and this effect showed a time-and concentration-dependency (r=0.94, 0.93, 0.92), the cell cycle was blocked in G0/G1 phase. As compared with control group, the proportion of Annexin V+PI-MUTZ-1 cells and the cellular PTEN levels increased in the treated group dramatically and in time-and dose-dependent manners (p<0.01). To the contrary, level of p-mTOR expression markedly decreased as compared with control group (p<0.05). It is concluded that the rapamycin inhibits the proliferation of MUTZ-1 cells, down-regulates the PTEN/PI3K-Akt/mTOR signaling pathway by interaction with mTOR, which induces the apoptosis of mUTZ-1 cells.

The correlation study of PTEN gene expression and Akt phosphorylation in myelodysplastic syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between PTEN gene expression and Akt phosphorylation (p-Akt) in myelodysplastic syndrome (MDS) and to explore the progression of MDS and the mechanism of high risk transformation to acute myeloid leukemia.</p><p><b>METHODS</b>RT-PCR was used to detect the PTEN mRNA expression in leukemia cell lines K562 (as negative control) and Jurkat (as positive control) and 65 MDS and MDS/AML patients. Flow cytometry was used to detect p-Akt in HL-60 and Jurkat cells and 30 MDS patients.</p><p><b>RESULTS</b>(1) K562 cells present PTEN gene expression while Jurkat cells did not. Of 65 MDS and MDS/AML patients, 27 (41.5%) expressed PTEN mRNA, being significantly lower than that in normal group (85.7%) (P < 0.01). (2) Jurkat cell showed high expression (86.9%) of p-Akt, while HL-60 cell as negative control did not express. P-Akt levels of 30 MDS patients were increased (1.35% - 58.23%), being much higher as compared with that of the normal contrast group (0.54% - 2.34%) (P < 0.01). Moreover, with the rate of blast cells increasing, the p-Akt level was rising up. There is a positive correlation (r = 0.93, P < 0.01) between the low expression rate of PTEN and the positive rate of p-Akt.</p><p><b>CONCLUSION</b>The loss of PTEN gene expression is one of the important factors of p-Akt high expression in MDS patients, moreover, it may speed up the progress of the MDS or transformation to acute myeloid leukemia.</p>

Expression of CD66c (CEACM6) in adult acute leukemia and its significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the expression of CD66c (CEACM6) in adult acute leukemia and its significance.</p><p><b>METHODS</b>Acute leukemia cell lines HL-60, K562, LCL721.221 and Jurkat were cultured in vitro. RT-PCR and multi-parameter flow cytometry were applied to analysis of CD66c mRNA and protein expression respectively in the cell lines and patient' s bone marrow leukemic cells. Cytogenetic analysis for 199 bone marrow samples from leukemia patients and Minimal Residual Disease (MRD) detection for 25 CD66c positive B lineage ALL were performed.</p><p><b>RESULTS</b>(1) CD66c expression both on cell surface and in plasma were negative in all the cell lines. (2) Four of 127 AML (3.15%) (mainly of M2 and M4), and 28 of 79 ALL (35.44%) (all of B linage ALL) were CD66c positive the subtypes of the ALL being common B-ALL (20/54) and pre B-ALL (8/11) including 8 Ph + B-linage ALL. (3) Six-month relapse rate was significantly different between the MRD positive and negative patients. (4) CD66c mRNA was strongly expressed in B-linage ALL. For the cell lines, only the HL60 cells weakly expressed CD66c mRNA.</p><p><b>CONCLUSION</b>CD66c expression could be a useful bio-marker for the MRD analysis in ALL, and is closely associated with its transcription level.</p>