Objective To study the lab-scale simulated method of three-effect dynamic countercurrent extraction from Hedyotis diffusa by multi-index optimization and SPSS software.Methods Polyphenol content,flavonoids content,scavenging activity of samples on DPPH radicals,and extracting rate were considered as the optimization indexes.Orthogonal design method was used to optimize the four main factors of alcohol concentration,ratio of solid to solution,extracting temperature,and extracting times.Results The optimum condition is 70 min extraction time,solid to solution ratio of 1∶14,50% alcohol,extracting temperature 85 ℃.Conclusion The condition of large-scale industrialized production might be explored by studying the lab-scale simulated method of dynamic countercurrent extraction from H.diffusa.

Cyclic peptide drugs have gradually become an emerging research direction due to their some favorable properties such as high-efficiency binding affinity, high selectivity, lower toxicity, and stable metabolism. In recent years, the number of cyclic peptide drugs under clinical research has continued to increase. Unlike the previous cyclic peptide drugs, which were mostly derived from natural products and their derivatives, these cyclic peptide drugs are designed by genetically encoded display technologies which are based on rational design and in vitro evolution (such as BT1718, PTG-300, POL6326, etc). Among them, phage display technology has some advantages such as mature research system, low cost, and simpler operation that make it well recognized and praised by the majority of researchers in this field. Here, we reviewed the recent progress of applying phage display technology to explore diverse cyclic peptide libraries, which, we believe, will contribute more valuable candidate cyclic peptide drugs in clinical research.

Bioimprinting is a new developed technique to improve the characteristics of enzymes.Bioimprinting by lauric acid was conducted to improve the esterification activity of lipase PS in sol-gel immobilization process with methyltrimethoxysila(MTMS) and tetramethoxysila(TMOS) as the precursors.Results generated by checking the esterification activity and scanning electron microscope showed that bioimprinting can enhance the specific activity and thermal stability of lipase PS.The bioimprinting system was optimized by orthogonal experiment,and the optimal condition for lipase bioimprinting is water/silane molar ration(R) 12,polyethylene glycol(PEG) 120?l,and lauric acid 0.15 mmol.Compared with the free enzyme and the non-imprinted enzymes,the specific activity of imprinted enzymes has been improved 44.3 fold and 2.4 fold,respectively.Imprinted lipase show better thermal stability,and the relative activity is 58% after incubated in 80 ℃ for 0.5 h,while no activity was detected for the free enzyme.

Objective To build a high quality diagnosis system for schistosomiasis surveillance in the situation of low infec-tion in Jianglin County. Methods The network laboratory for schistosomiasis diagnosis was built according to the national crite-ria in Jianglin County in 2012. Results The network laboratory for schistosomiasis diagnosis was established successfully and the operation was quiet well. Conclusion The establishment and operation of the laboratory play an important role in the real-ization of schistosomiasis elimination.

OBJECTIVETo explore the relationship between microRNA and drug resistance in leukemia treatment by screening and identifying the microRNAs which differentially express in K562 cell line and its adriamycin resistant cells-K562/A02 cell line.

METHODSThe drug resistance potency of K562/A02 cells was evaluated by MTT assay. P-gp expression of K562 and K562/A02 cells were detected by flow cytometry (FCM). The differentially expressed microRNAs in K562 and K562/A02 cells were analyzed by microarray technique and Real Time RT-PCR.

RESULTSThe resistance to adriamycin (ADM) of K562/A02 cells was 180 fold greater than that of K562 cells. P-gp expression rate of K562 and K562/A02 cells was 0.2% and 86%, respectively. Twenty-two microRNAs expressed differentially in K562 and K562/A02 cells (P < 0.01). As compared to K562 cells, expressions of miR-221, miR-155 and miR-451 were up-regulated by more than two fold, while expression of miR-98, miR-181a, let-7f, let-7g, miR-424 and miR-563 down-regulated by more than two fold in K562/A02 cells. The results of real time RT-PCR were consistent with that of microarray. Of note, differential expressions of miR-451, miR-155, miR-221, let-7f and miR-424 were remarkable.

CONCLUSIONK562/A02 cells show a different microRNA expression profile as compared to its parental K562 cells, suggesting microRNAs including miR-221, miR-155, miR-451, let-7f and miR-424 may be involved in the mechanism of drug resistance in leukemia. These differentially expressed microRNAs provide potential novel targets for overcoming drug-resistance.

Doxorubicin ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; MicroRNAs ; genetics

OBJECTIVETo study the chemical constituents of Daphne odora var. margirmta.

METHODThe compounds were isolated and purified by column chromatography of silica gel, ODS, Sephadex LH-20, and their structures were identified by UV, 1H-NMR, 13C-NMR, MS and CD spectroscopic methods.

RESULTTen compounds were elucidated as wikatrols B (1), kaempferol (2), daphnodorin B (3), daphnodorin D2 (4), daphnodorin A (5), daphnodorin C (6), daphnodorin D1 (7), daphnodorin I (8), daphnetin (9), daphnoretin (10).

CONCLUSIONCompounds 1,2,4 were obtained from this plant for the first time,and compound 1 was isolated firstly from the genus Daphne.

Daphne ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plant Roots ; chemistry

Objective To investigate the value of three-dimensional speckle tracking imaging (3D-STI) in assessment of left ventricular (LV) strains. Methods Thirty healthy young adults examined by two-dimensional speckle tracking imaging (2D-STI) and 3D-STI. And the results of LV measurements were compared, which included mean peak systolic longitudinal strains, radial strains and circumferential strains. Also, the time consumption of these two methods was compared. Results The time needed for 3D-STI in acquisition and analysis of the images were (309.3±23.4)s, (305.5±11.2)s, while the time for 2D-STI were (490.6±14.4)s, (1261.4±39.9)s. The differences were signiifcant(t=-21.81, 69.94, both P<0.01). The global mean peak systolic radial strains was (48.59±7.68)%by 3D-STI and (33.25±7.27)%by 2D-STI. The difference was signiifcant(t=9.16, P<0.01). The global mean peak systolic longitudinal and circumferential strains were (-17.66±3.14)%, (-17.13±2.29)% by 3D-STI and (-21.35±2.46)%, (-21.97±3.84)% by 2D-STI. The differences were signiifcant(t=5.33, 5.99, both P < 0.01). The 3D-STI strains were different at different levels of LV. The longitudinal, circumferential and radial 3D-STI strains were largest at middle levels. However, 2D-STI strains didn′ t show such trend. Peak strains measured by 3D-STI and 2D-STI showed high inter-observer and intra-observer agreement in Bland-Altman chart. Conclusion 3D-STI is a novel, convenient and reproducible method to evaluate the strains of LV.

Objective To evaluate the efficacy of routinely used pattern for schistosomiasis diagnosis in lake and marshland regions. Methods A historically heavy endemic village of schistosomiasis named Jinggan Village from Jiangling County was se?lected for field survey. The residents aged 6?65 years were screened by indirect hemagglutination assay(IHA)and dipstick dye immunoassay(DDIA)in parallel. The serological positives were examined by Kato?Katz technique and miracidium hatching technique to determine the infection of schistosome. The consistency of the two serological methods was evaluated. In addition , the schistosome infection rates were estimated according to the 3 detection patterns namely IHA,DDIA,IHA+DDIA combined with the etiologic examination. Results A total of 530 individuals were examined by the serological tests. The positive rate of DDIA was 46.98%(249/530),significantly higher than that of IHA(28.49%,151/530)(χ2=59.55,P<0.01). Totally 279 in?dividuals were serological positives determined by IHA or DDIA,while 252 of them were detected by stool examination,and 22 cases were determined as parasitological positives,while 7 and 3 cases were diagnosed as antibody negatives by IHA and DDIA,respectively. The estimated infection rates determined by IHA,DDIA,IHA plus DDIA combined with stool examination were 3.14%,3.97%、4.60%,respectively. Conclusions Under the condition of endemic situation becoming more and more waning,the current routinely used pattern for schistosomiasis detection may lead to missed diagnosis. So,more sensitive and ef?fective diagnostic tools or appropriate detection patterns need to be explored.

OBJECTIVETo investigate the effects of acute heat stress on the expression patterns of heat shock protein 70 (HSP70) in the testis, epididymis and vas deferens of adult male mice.

METHODSThirty-two 8-week-old male mice were randomly divided into a control and 3 heat stress groups. After a week of pretreatment, the latter 3 groups were exposed to heat stress at (39 +/- 0.5) degrees C for 0.5, 1 and 3 hours, respectively, followed immediately by collection of the blood and determination of glutamic oxaloacetic transaminase (GOT) concentration. Sperm suspension was made from one epididymis for the detection of sperm concentration and abnormal acrosome rate, while the testis, epididymis and vas deferens on the other side were harvested for immunohistochemical analysis.

RESULTSHeat stress induced different degrees of decrease in epididymis indexes and sperm concentration and a dramatic increase in GOT concentration (P < 0.01), but caused no significant changes in the body weight, testis indexes and abnormal acrosome rate in the mice (P > 0.05). There was a tendency of gradual descent in sperm concentration and ascent in abnormal acrosome rate with the increasing time of heat stress. The most decrease in body weight, testis indexes and epididymis indexes was observed in the 0.5 h group. Immunohistochemical results showed that HSP70 was expressed in the testis, epididymis and vas deferens, mildly in the Leydig cells of the testis and distributed in the nuclei of the Leydig cells, spermatoblasts and spermatocytes. In the epididymis, HSP70 was expressed mainly in the cytoplasm of principal and ciliated cells, but not in the basal and clear cells, while in the vas deferens, it was observed mainly in the cytoplasm of the basal cells, but not in the principal cells. With the increase of heating time, the HSP70 expression was markedly elevated in the testis and epididymis, but not so obviously in the vas deferens.

CONCLUSIONAcute heat stress was harmful to the reproductive system of adult male mice. The region- and cell-specific patterns of HSP70 expressions in the testis, epididymis and vas deferens suggested that HSP70 might play an important role in spermatogenesis and sperm maturation. The dramatic increase of HSP70 expression after heat stress might be responsible for the prevention of cells from high temperature.

Animals ; Epididymis ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Heat Stress Disorders ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Testis ; metabolism ; Vas Deferens ; metabolism

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.