To explore the effect of quercetin on the proliferation and apoptosis of HeLa cells, HeLa cells were incubated with quercetin at different concentrations. Cell viability was evaluated by MTT assay, cell apoptosis was detected by Annexin-V/PI double labeled cytometry and DNA ladder assay. Cell cycle was flow cytometrically determined and the morphological changes of the cells were observed under a fluorescence microscope after Hoechst 33258 staining and the apoptosis-related proteins in the HeLa cells were assessed by Western blotting. The results showed that quercetin significantly inhibited the growth of HeLa cells and induced obvious apoptosis in vitro in a time- and dose-dependent manner. Moreover, quercetin induced apoptosis of HeLa cells in cell cycle-dependent manner because quercetin could induce arrest of HeLa cells at G0/G1 phase. Quercetin treatment down-regulated the expression of the PI3K and p-Akt. In addition, quercetin could down-regulate expression of bcl-2, up-regulate Bax, but exerted no effect on the overall expression of Akt. We are led to conclude that quercetin induces apoptosis via PI3k/Akt pathways, and quercetin has potential to be used as an anti-tumor agent against human cervix cancer.

Objective To determine the effect of endomorphin (EM) on colonic electromyography activity and investigate the pathogenesis of slow transit constipation (STC). Methods An experimental rat model of slow transit constipation was constructed by contract laxatives mixed with the feed. The changes of colonic electromyography and reaction to endomorphin 1 and endomorphin 2 were examined. Results Compared with the control group, the frequency and amplitude of slow wave in cathartic colon rats were decreased significantly. Endomorphin 1 and endomorphin 2 significantly decreased the amplitudes of slow wave, but did not change the frequencies of slow wave. The effect of endomorphin 1 was more pronounced than that of endomorphin 2, which could be reversed by the morphine antagonist Naloxone in concentration-dependent manner. Endomorphin could not block the stimulating effect of acetylcholine. Conclusions Endomorphin can influence the colonic electromyography activity and intestine motility of cathartic colon rats, and may be involved in the pathologic mechanism of slow transit constipation.

Adolescent ; Child ; Humans ; Male ; Neuroectodermal Tumors, Primitive, Peripheral ; pathology ; Sarcoma, Ewing ; pathology

Cerebral Palsy ; mortality ; pathology ; physiopathology ; Child ; Humans ; Prognosis ; Survival Analysis ; Survival Rate ; Time Factors ; Walking

Diagnosis, Differential ; Humans ; Hyponatremia ; diagnosis ; etiology ; therapy ; Inappropriate ADH Syndrome ; diagnosis ; etiology ; therapy

Objective: The calibration models were developed in the concentration of alcohol precipitation proee for Artemisiae Annuae Herba (AAH) and Lonicerae Flos (LF) in Reduning Injection (RI) to realize the on-line monitoring of production process. Methods: Based on the near infrared reflectance spectroscopy (NIRS), partial least regression (PLS) models were developed to fast measure the contents of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid in the concentration of the alcohol precipitation proee for AAH and LF. Results: In the quantitative models of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid, the coefficient of determination (R2) of cross validation sets were 0.954 5, 0.975 2, and 0.969 1; The root mean square errors of calibration (RMSEC) were 0.213, 0.676, and 0.225; The root mean square errors of cross-validation (RMSECV) were 0.233, 0.692, and 0.258. When the established models were applied to on-line monitoring, the coefficient of determination of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid were 0.984 2, 0.983 7, and 0.987 0, the residual predictive deviation (RPD) were 4.77, 5.29, and 4.37; The relative standard errors of prediction (RSEP) were 3.519%, 3.778%, and 3.895%. Conclusion: The models above are proved to fast measure the contents of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid in the concentration of alcohol precipitation proee for AAH and LF in RI.

Objective:To study the application of statistical process control technology to establish the real-time release criteria of alcohol precipitation process of ArtemisiaeAnnuae Herba and Lonicerae Flos. Methods: Twenty-ninebatches of samples came from alcohol precipitation process of ArtemisiaeAnnuae Herba and Lonicerae Flos were collected as the calibration set. The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, and solid content were determined to establish the quantitative real-timerelease criteria based on statistical process control technology. Thirteenbatches of alcohol precipitation process of ArtemisiaeAnnuae Herba and Lonicerae Flos were prepared under the different process conditions by the central composite design. They were regarded as the validation set to test the dependability of the real-time release criteria. Results: The established quantitative release ranges were neochlorogenic acid 0.279-0.541 mg/g, chlorogenic acid 1.941-2.610 mg/g, cryptochlorogenic acid 0.453-0.570 mg/g, and solid content3.565%-4.925%. The four index component contentsof the samples 1, 4, 8, 9, and 10 from the validation set werewithin the range of the real-time release criteria. Conclusion: Univariate statistical process control could be well monitored and understood of production process for Chinses materiamedica.They are used to achieve the purpose of real-time release.

To optimize the alcohol precipitation process for Lonicerae Flos and Artemisiae Annuae Herba (LA) in Reduning Injection and obtain the relationship equation of the key process parameters and quality attributes, which could provided a theoretical basis of the control automization in the alcohol precipitation process. With the transfer rates of neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, caffeic acid, secoxyloganin and solid content in precipitation liquid as evaluation indexes, the effect of the seven factors, such as relative density and the temperature of LA in liquid phase before alcohol precipitation, alcohol concentration in the end of alcohol precipitation, stirring speed in alcohol precipitation, adding alcohol speed, standing temperature, and standing time, on the alcohol precipitation process was investigated by the single factor experiment. By analysis of variance, the key factors that could influence the alcohol precipitation process were determined. Then the range of parameters of key factors was further studied and explored by Box-Behnken response surface methodology. The optimum preparation conditions of the alcohol precipitation process of LA were as follows: stirring speed was 550 r/min, adding alcohol speed was 4.0 mL/s, standing temperature was 30℃, standing time was 24 h, alcohol concentration was 75%, the relative density of LA was 1.10, and the temperature of LA was 25℃. Under these conditions, the transfer rates of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, and secoxyloganin were 94.8%, 97.6%, 97.4%, 97.2%, and 96.1%, and the solid content was 4.2%. The alcohol precipitation process of LA have been optimized by the response surface method, which could help to enhance the stability of the process.

Objective To observe the effect o f selective prostaglandins EP 1 receptor antagonist SC19220 on the afferent activity of C unit in the saphenous nerve of diabetic rats. Methods The response of C units to sustained 1 min threshol d and suprathreshold stimuli was measured following intradermal injection of EP 1 receptor antagonist SC19220 into the receptive field of C unit of the sa phenous nerve in control and diabetic rats. Results The mechanical threshold of C units in diabetic rat s was obviously decreased compared with that of control rats (P

Background and purpose:The theory of cancer stem cell offers us a new thought about tumors, more and more kinds of cancer stem cell were isolated and indentif ied from corresponding cancer tissue. Our aim was to investigate the content of side population cells in SW480 human colorectal cancer ce11 line and to enrich cancer stem- like cells in SW480 through serum-free medium (SFM) culture. Methods:The percentage of side population cells in human colorectal cancer cell line SW480 was detected with ? ow cytometry. SW480 cell line was cultivated in serum- free medium(SFM) supplemented with growth factors and the cancer stem-like cells reforming into ? oating spheres were isolated. The isolated cancer stem-like cells were identifi ed by limited-dilution assay, differentiation assay, self- renewal assay, and alternative cultivation assay. Results:The percentage of SP cells was 1.2% in SW480,In the absence of serum, a minority (0.54%-0.62%) of cancer stem-like cells in SW480 cells survived, proliferated and formed into the suspended tumor cell spheres. SW480 cancer stem-like cells possessed proliferative, self-renewal and differentiation potential, which were responsible for the ? oating tumor clone. Serum addition into SFM resulted in the proliferation of cancer stem-like cells; after several generations and alternated cultivation in SSM and SFM, cancer stem-like cells maintained their characteristics. Conclusions:SW480 cell line contains a tiny minority of SP cells with stem cell properties.The cancer stem-like cells in SW480 line can be maintained in SFM using a floating culture method.