Objective To study ischemic tolerance induced by acanthopannx senticosus saponins(ASS) on ischemia in PC12 cells and involve expression of hypoxia inducible factor-1?(HIF-1?).Methods An ischemic model was developed in PC12 cell line with treatment of oxygen glucose deprivation(OGD).The protective effects of ASS pretreatment on ischemic tolerance of PC12 cells and whether protective effects of ASS could be inhibited by LY294002(PI3K inhibitor) were analyzed through MTT assay.The induction of phospho-glycogen synthesis kinase-3?(p-GSK-3?) and HIF-1? after pretreatment of ASS and possible blocking effects of LY294002 were detected by Western-blot.Results MTT results showed that after 9 h ischemia,the viability of PC12 cells decreased dramatically(P

Objective To detect the excision repair cross-complementing gene 1 (ERCC1) and thymidylate of the acid synthase (TS) in non-small cell lung cancer (NSCLC) and its adjacent tissue,and investigate the relationship of the expression of ERCC1 and TS with the clinical characteristics of NSCLC and prognosis for NSCLC individual therapy to provide experimental basis.Methods The protein expression levels of ERCC1 and TS in 50 cases of postoperative NSCLC cancer and adjacent tissue were detected by immunohistochemical method and the relationship among the expression of ERCC1,TS,and overall survival of patients with NSCLC Phase (OS),disease progression time (TTP),the median OS,and median TTP was analyzed.Results (1)There was an obvious difference between the expression of ERCC1,TS in cancer and paraneoplastic tissue of NSCLC,which had statistically significance (64.00％ vs 20.00％,x2 =19.87,P ＜ 0.01 ;48.00％ vs 24.00％,x2 =6.25,P ＜ 0.05) ; (2)The continued investigation in the patients who received postoperative cisplatin or carboplatin chemotherapy showed that the 0S of negative expression of ERCC1 was significantly longer than the positive one (19.10 vs 10.00 months;x2 =8.133,P =0.002),so was median TTP (15.30 vs 9.00 months; x2 =7.410,P =0.003).The median 0S of the negative expression of TS,was significantly longer than the positive one (17.80 vs 11.00 months,x2 =7.001,P =0.008),so was median TTP (11.40 vs 6.80 months; x2 =5.884,P =0.026).Conclusions ERCC1 and TS protein may become sensitive predictors of platinum chemosensitivity for NSCLC patients ; the detection of combined with ERCC1 and TS would contribute to the selection of individualized treatment programs for NSCLC.

Objective To evaluate left ventricular global longitudinal strain in endocardium,middle myocardium and epi cardium of patients with breast carcinoma treated with anthracyclines by two-dimensional layered strain technology.Methods Totally 20 breast cancer patients after surgery without chemotherapy (control group) and 32 breast cancer patients who were treated with anthracycline-based chemotherapy were enrolled.The patients treated with chemotherapy were performed transthoracic echocardiography 3 months and 5 months after chemotherapy.Global longitudinal strain (GLS) and each segment (apex,basis and media section) strain in 3 myocardial layers (endocardium,middle myocardium and epicardium) were measured by EchoPAC software.Results Compared with control group,with the extension of chemotherapy cycle,GLS of 3 myocardial layers reduced in breast carcinoma patients treated with anthracyclines (all P＜0.05),but left ventricular ejection fraction (LVEF) had no statistical difference (P＞0.05).Compared with control group,each layer of each segment of longitudinal strain decreased in breast carcinoma patients treated with anthracyclines (all P＜0.05),except the apex section.Conclusion Longitudinal layer-specific strain technology can accurate assess whole and local systolic function of 3 myocardial layers of left ventricular,which can provide a new method for judging myocardial damage.

Objective To study the relationships between hepatocellular carcinoma (HCC) and the polymorphisms,promoter methylation,and expression of glutathione S-transferases P1 gene (GST) P1 gene.Methods Using methylation-special PCR (MSP),the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied.The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry.And using PCR-RFLP,the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied.Results The diffe-rences of the frequency of GSTP1 Ile/Ile,Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance (X~2=0.84,v=2,P=0.656).The frequency of methyla- tion of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when com- pared to the corresponding nontumor tissues (X~2=19.08,P＜0.001),and significantly higher in stageⅢ-Ⅳcases when compared to the stageⅠ-Ⅱcases (X~2=4.84,P=0.028).GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues (t=2.49, P=0.014),and significantly higher in stageⅠ-Ⅱcases when compared to the stageⅢ-Ⅳcases (t= 2.31,P=0.025).On the other hand,the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation (t=3.50,P=0.001).Conclusion GSTP1 inactivation via CpG island hypermethylation may contrib- ute to the pathogenesis of HCC.

To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 μg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drug Contamination ; Mycotoxins ; analysis ; Panax notoginseng ; chemistry ; Tandem Mass Spectrometry

BACKGROUND:A variety of embryonic stem cells induction and differentiation systems have been established so far, while the research that promotes embryonic stem cells to differentiate into hematopoietic stem cells is stil at an initial stage, and the induction efficiency needs to be improved.
OBJECTIVE:To active the Wnt/β-catenin signal pathway in mouse embryonic stem cells with exogenous win3a as an inducer, and then to observe whether the activation of this pathway wil promote the directional differentiation of embryonic stem cells into hematopoietic progenitor cells.
METHODS:The ES-E14TG2a mouse stem cells were cultured with the exogenous wnt3a (100 μg/L) for 21 days, the content ofβ-catenin was tested by cellimmunofluorescence and western blot, and expression of Wnt downstream target gene was detected by quantitative reverse transcription PCR to determine the activation of Wnt/β-catenin signal pathway. Single-layer adherent culture method was used to induce the directional differentiate of above-mentioned cells into hematopoietic stem cells, and detection of hematopoietic development associated surface marker CD34+/Sca-1+was achieved by flow cytometry;meanwhile, the expression of hematopoietic associated gene was measured by quantitative reverse transcription PCR.
RESULTS AND CONCLUSION:We found thatβ-catenin accumulated in ES-E14TG2a mouse stem cells after cultured with wnt3a (100 μg/L) for 21 days;the expressions of Wnt downstream target genes such as Pitx2, Frizzled, Sox17 and Oct4 showed the different degrees in increase, meaning the activation of Wnt/β-catenin signal pathway. Furthermore, during the time that we used single-layer adherent culture method to induce hematopoietic stem cells, the CD34+/Sca-1+cells accounted for 20.2%of total cells at day 14, and control cells only accounted for 11.9%. Again, expression quantity of hematopoietic associated gene BMP4, FLK2 and CD34 increased while Smad5 was suppressed significantly. Our data suggest that sustaining action by wnt3a wil active Wnt/β-catenin signal pathway, and also promote the directional differentiation of ES-E14TG2a mouse stem cells into hematopoietic progenitor cells.

Objective To explore the mechanism of the referred pains in pelvic pain due to chronic prostatitis and the relation between pain due to prostatitis and pelvic floor muscle dysfunction. Methods The retrograde fluorescent double labeling and immunohistochemistry were used. A total of 24 Wistar rats were divided two groups, 12 in each group. In the first group, Propidium Iodide(PI) was injected into the prostate, Bisbenzimide(Bb)into the external anal sphincter. In the second group, PI was injected into the prostate, Bb into genitofemoral, iliohypogastric, ilioinguinal, femoral and lateral femoral cutaneous nerves. Results The fluorescent double labeled cells were found in the L 1 L 2 and L 6～S 3 dorsal root ganglia(DRG) and some of them were confirmed to contain calcitonin gene related peptide(CGRP) and P substance (SP) by immunohistochemistry. Conclusion The findings confirm that the peripheral process of DRG cells dichotomizes into prostate, external anal sphincter and somatic parties, and some of these cells contain CGRP and SP, which indicates the referred pains in pelvic floor may be caused by axon reflex in the peripheral process of DRG cells. Meanwhile, neurogenic inflammation might play an important role in this persistent pain status. Pain due to prostatitis is correlated with pelvic floor muscle dysfunction.

Objective To elucidate the neurological mechanism of the referred pains in pelvic floor associated with the prostate. Methods The rat prostate was electrically stimulated, then the elicited action potentials (AP) of the lumbar and sacral nerves and the compound muscle action potentials from the external sphincter muscle of anus were detected. Results Action potentials from the iliohypogastric, genitofemoral, lateral femoral cutaneous, femoral, sciatic nerves compound muscle action potentials (CMAP) from the anal external sphincter were recorded. Conclusion Electric stimulation of the prostate can induce action potentials of the lumbar and sacral efferent nerves through the spinal reflex, resulting in referred pains in the perineal and pelvic areas due to muscle contraction.

Purpose To observe the effect of acupuncture on gnosia and acetylcholinesterase in patients with senile dementia. Methods Eight patients diagnosed with mild or moderate senile dementia were treated by acupuncture of Sishencong ( Ex-HN 1 ), Shenmen ( HT 7) and Taixi ( KI 3) for I month; gnosia was evaluated by Mini-mental state examination before and after the treatment; plasma acetylcholin esterase activity was measured by flourier before and after the treatment. Results There was a significant difference in gnosia between pre- and post--treatment with acupuncture (P＜0.01); there was no significant difference in acetyl- cholinesterase activity between pre- and posttreatment ( P＞0.05 ). Conclusion Acupuncture has a certain improving effect on gnosia in senile dementia;one month's acupuncture treatment had little effect on plasma acetyl-cholinesterase activity.

Objective:To explore the characteristics of human melanoma-specific antigen peptides by HLA-A2 restricted tumor-infiltrating lymphocytes.Methods:The HLA-A2 protein and polypeptides molecules were purified from the three tumor cell lines(624-Mel, Chap-Mel and JY) by immunoaffinity chromatography, after the peptides bound to HLA-A2 protein solution were acidified with acetic acid and boiled by high temperature, and centrifuged through an Ultra-CL filter, then the peptides extracts were fractionated by revered phase high pressure liquid chromatography(RP-HPLC). Individual fractions were assessed for their ability to reconstitute melanoma-specific epitopes by adding to the HLA-A2 Ag-procceing mutant cell, T2. The biological feature of one of three active peptides from RT-HPLC samples was performed by mass spectrometric analysis. The synthetic peptides identical to active peptide sequences were determined in the reconstitute test.Results:Three prominent peaks(P19, P25 and P31) of the fraction from 624-Mel were observed in the reconstitute test, TIL killing rate was 67% for (P31) peptide fraction. The mass spectrometric analysis of one of active peptides (P31) showed that at mass-to-charge ratio(m/z) 948 has been usually nine residues. The sequence is H+ Ala Lue Trp Lue Phe Phe Gly Val Lue OH-. The peptide synthesized comprising epitopes were verified.Conclusion:These results showed the peptides derived from active fractions were related to human melanoma-specific tumor antigen peptides recognized by HLA-A2-restriced TIL. These peptides could develop novel peptide-based an anti-tumor vaccine for immunotherapy of CTL.