An efficient technique for the determination of multiclass pesticide residues ( glyphosate, aminomethyl phosphonic acid ( AMPA) and glufosinate) in four kinds of tea ( green tea, black tea, Oolong tea and Puer tea) was developed based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were extracted with 0. 05 mol/L NaOH solution, regulated pH value with HCl, followed by purification by Oasis HLB column, and then precolumn-derived with 9-fluorenylmethyl chloroformate ( FMOC-CL ) in borate buffer. All pesticide residues studied showed good linearity with correlation coefficient ( R2 ) greater than 0. 99 in the concentration range of 5-1000 μg/L. Limits of quantitation (LOQs) of three pesticide compounds ranged from 0. 03 to 0. 08 mg/kg (S/N=10). Tea samples spiked with 0 . 1 , 0 . 4 and 4 mg/kg showed the recovery ranging from 72 . 1% to 109 . 9%, and RSDs from 0. 5% to 9. 8% (n=6). In addition, the validated method was applied to commercial samples, and all the detections were confirmed by acquiring transitions for each pesticide in samples.

Objective To investigate the effect of DHA on proliferation and killing activity of γδT cells against SW1990 cells in vitro.Methods γδT cells were generated in vitro by stimulating peripheral blood mononuclear cells of healthy donors in RPMI 1640 completed medium containing IPP and IL-2 for 8 d,and then co-cultured with different concentrations of DHA for 48 h.Proliferation rates of γδT cells for each group were detected by MTT method.The perforin,granzyme B and CD107a expression in γδT cells were verified by flow cytometer.The cytotoxic activity of γδT cells against SW1990 cells were analyzed by CCK-8 kit.Results The purity of γδT cells in each group reached 75.46％ ±5.32％ after 8 d of culture.Compared with the control group,the proliferative capability of γδT cells were enhanced significantly after treated with 50-100 μmol/ml DHA for 48 h,moreover,cytotoxicity against SW1990 cells and perforin,granzyme B and CD107a expression of the γδT cells treated with DHA were higher than the control group.Conclusion DHA could enhance the antitumor activity of γδT cells,which may be associated with the upregulation of perforin and granzyme B expression in γδT cells.

Purpose:To study the expression of COX 2,P53 and PCNA in esophageal carcinomas and its significance. Methods:Immunohistochemical method was used to examine the sections from 82 esophageal squamous cell carcinomas, 20 esophagitis and 16 normal esophageal mucosa tissues. Results:The positive ratios of COX 2,p53 and PCNA were 87.8%(72/82),82.9%(68/82)and 95.1%(78/82) in 82 esophageal carcinomas, respectively. But there was no expression in adjacent noncancerous tissues and normal esophageal mucosa tissues. The positive ratio of COX 2 protein was significantly higher in the well differentiated and moderately differentiated esophageal carcinomas than in the poorly differentiated esophageal carcinomas( P

Background and purpose: Human epididymis protein 4 (HE4) has been suggested to be a novel biomarker of epithelial ovarian cancer (EOC). The present study aimed to evaluate and compare HE4 with the commonly used marker, carbohydrate antigen 125 (CA125) and imaging examinations in detecting the recurrence of postoperative EOC. Methods:In this study, 92 patients with the recurrence of postoperative EOC were analyzed retro-spectively. The treatments method included the secondary operation (48 patients) and chemotherapy (44 patients). The sensitivity of CA125, HE4 and CT/MRI examinations in detecting the recurrence of postoperative EOC were analyzed and correlated with surgical pathology and clinical follow-up results. Results:The sensitivity of CA125 and HE4 were 58.7%and 61.9%with no signiifcant difference (P>0.05). The diagnostic sensitivity of CA125 combined with HE4 (80.4%) was statistically higher than that of 2 tumor marks being applied independently (P>0.05). The diagnostic sensitivity of CT/MRI examinations (88.0%) was statistically higher than that of CA125 and HE4 (P<0.05), and no signiifcant difference was observed between CT/MRI and combined CA125 and HE4 (P>0.05). The highest diagnostic sensitivity (97.8%) was attained by combining of CA125, HE4 and CT/MRI examinations, which was statistically high-er than that of CA125 combined with HE4 (P<0.01), and CT/MRI examinations (P<0.05). Conclusion:Serum HE4 is an effective tumor marker in detecting the recurrence of postoperative EOC as well as serum CA125. HE4 and CA125 may improve the diagnostic sensitivity statistically. The highest diagnostic sensitivity was attained by combining of CA125, HE4 and CT/MRI examinations, which is the suitable strategy in screening the patient of postoperative EOC.

Objective To investigate the effects of hydrocortisone (HC) on proliferation and killing activity of NK cells against pancreatic cancer SW1990 cells in vitro.Methods Peripheral blood mononuclear cells of healthy people were isolated and cultured with NK cells medium containing IL-1S.When the purity of NK cells reached above 70％,different concentrations of HC (10-6,10-5,10-4,10-3 μmol/L) were added and co-cultured with NK cells for 7 days.And NK cells without HC were used as control.CD3-CD56 + NK cell numbers of each group were countered by trypan blue staining.Perforin,granzyme B and IFN-γ expression of CD3-CD56+ + NK cells were verified by flow cytometry.NK cells and SW1990 cells were co-cultured with a 20∶1effector to target ratio,then the cytotoxic activity of NK cells against SW1990 cells were analyzed by CCK-8 kit.Results After treatment with different concentration of HC for 7 days,NK cells purity of each group reached 70.72％ ～ 76.39％,and it was not significantly different with that in control group [(72.61 ± 3.76) ％].The proliferation folds of NK cells treated with 10-6,10-5,10-4,10-3 μmol/L HC were (9.13 ± 0.94),(9.67 ±1.51),(10.33±1.07),(8.40±1.47) times,respectively,while it was (4.23 ±0.82) times in control group (all P ＜0.01).The killing effects of NK cells on SW1990 cells were (58.58 ± 4.89) ％,(62.27 ± 5.63) ％,(64.02 ± 5.79) ％,(63.88 ± 3.61) ％,which were higher than that in control group [(57.46 ± 5.11) ％],moreover,the difference between NK cells of 10-4 μmol/L HC treatment group and control group was statistically significant(P ＜ 0.05).The expressions of perforins of 10-4,10-3 μmol/L HC treatment group were (96.71 ± 3.04) ％,(97.56 ± 2.18) ％,which were significantly higher than that in control group [(92.40 ±3.53)％,P ＜0.05 or 0.01].The expression of granzyme B in 10-5 μmol/L HC treatment group was (78.23 ±2.94)％,which were significantly higher than that in control group [(73.68 ±3.52) ％,P ＜0.05].The expressions of IFN-γ in 10-5,10-4,10-3 μmol/L HC treatment group were (96.61 ±2.04)％,(97.58 ± 2.17)％,(98.00 ± 1.77)％,which were significantly higher than that in control group [(92.44 ± 2.74)％,P＜0.01].Conclusions HC can promote IL-15 activated NK cells proliferation and enhance NK cells mediated killing activity against SW1990 cells with proper concentration,and up-regulation of perforin,granzyme B and IFN-γ expression may be the main mechanisms.

Objective To optimize the preparative procedure for stachydrine in Fructus Leonuri. Methods The preparation was screened by orthogonal experiment, and a mathematical model of relationship of extraction time, methanol concentration, and solid-liquid ratio with the content of stachydrine hydrochloride was established by using back-propagation (BP) neural network. And the process parameters were optimized with genetic algorithm (GA) . Results The optimum process parameters were as follows: extraction with 69% of methanol concentration and with solid-liquid ratio being 11 times for 62 min. The content of stachydrine obtained by BP neural network modeling and GA was higher than that achieved by orthogonal experiment. Conclusion The optimum preparative procedure could be achieved by combining BP modeling with GA. The model developed in this study was proved to be predictable and feasible for the optimization of process parameters of multi-dimension nonlinear system.

An efficient method for the analysis of multiclass plant growth regulators and pesticide (imidacloprid, acetamiprid) residues in tea was developed based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were extracted with acetonitrile/formic acid (99∶1, V/V) solution, cleaned up with four sorbents including C18, strong anion exchanger (SAX), primary secondary amine (PSA) and anhydrous MgSO4. The compounds were separated on a HSS T3 column under positive/negative electrospray ionization mode, detected by scheduled multiple reaction monitoring (SMRM), and quantified by matrix-matched external standard curves. All pesticide residues showed good linearity in the concentration range of 1-200 μg/L (6-benzylaminopurine, paclobutrazol, uniconazole, forchlorfenuron, mepiquat chloride, imidacloprid, acetamiprid) or 5-1000 μg/L (2,4-dichlor-ophenoxyacetic acid, 4-chlorophenoxyacetic acid, indole-3-acetic acid, gibberellic acid, 1-naphthaleneacetic acid, indole-3-butyric acid) , with correlation coefficient (R2≥0.99). Limits of detection (LOD, S/N=3) and limits of quantitation (LOQ, S/N=10) were 0.18-9.68 μg/kg and 0.61-32.26 μg/kg, respectively. In addition, the spiked recoveries of tea samples were 73.1％-108.9％, and RSDs were 0.6％-8.0％. This method was applied to commercial samples, and all the detections were confirmed by acquiring transitions for each pesticide in the samples.

Objective:To explore the mechanism of the cytotoxicity of human NK cells induced by atorvastatin to colon cancer cell lines. Methods:After colon cancer cells (HCT-116,SW-480,Caco-2) were cultured with different concentrations of atorvastatin, CCK-8 assay was used to assess the effect of atorvastatin on growth of colon cancer cells. The amplification of human NK cells was induced by SCGM medium in vitro. Automatic biochemical analyzer was applied to test the cytotoxicity of NK cells to colon cancer cells which cultured with different concentration of atorvastatin. FCM was used to detect the expression rate of MICA/B on the cells. Results:(1) The cultivation of NK cells:The proportion of NK cells attained to 93. 1% from 4. 5% after cultured for 10 days. (2) The effects of atorvastatin on the growth of the colon cancer cells:After cultured with atorvastatin,the inhibition rate of HCT-116 cells was higher than that in control when the density of atorvastatin increased from 5 μmol/L to 40 μmol/L after 48 h and from 1. 25 μmol/L to 40 μmol/L after 96 h ( P<0. 05 ) . Correlation analysis showed that the concentration of atorvastatin and the growth inhibition rate of HCT-116 cells were positively correlated(r[48 h]=0. 13,r[96 h]=0. 22,P<0. 05). (3) The cytotoxicity of NK cells to colon cancer cells effected after atorvastatin: In different atorvastatin concentrations groups,the cytotoxicity of NK cells to three colon cancer cell lines was all higher than that in control ( P<0. 05 ) . The atorvastatin concentration was from 2. 5 μmol/L to 10 μmol/L for HCT-116 cells,from 5 μmol/L to 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 20μmol/L for Caco-2 cells. Among the three cell lines, the cytotoxicity of NK cells to HCT116 was the highest in the same concentration. (4)NK cells by atorvastatin cutting statins 96 h,the concentration of 20 mmol/L and 40 mmol/L inhibition rate was higher than that of control group,more than other groups on NK cell growth without significant effect. ( 5 ) The impact of atorvastatin on MICA/B expression of colon cancer cells: After cultured with different concentrations of atorvastatin,the expression of MICA/B on colon cancer cells was higher than that in control(P<0. 05). The concentration was 2. 5μmol/L and 5μmol/L for HCT-116 cells,10μmol/L and 20μmol/L for SW-480 cells,and from 2. 5μmol/L to 40 μmol/L for Caco-2 cells. Conclusion:Atorvastatin could inhibit the growth of colon cancer cells (HCT-116,SW-480 and Caco-2) in a dose-dependent manner;and it could enhance the cytotoxicity of NK cells to colon cancer cells;it also could promote the expression of MICA/B of colon cancer cells,and improve the immunogenicity of colon cancer cells.

Objective To study the effect of Bairui granule and Deacetylated Ligustrazin injection on plasma BNP and FABP levels in children with heart failure and pneumonia.Methods 60 children of heart failure and pneumonia who received therapy from July 2014 to June 2016 in our hospital were randomly divided into two groups, the observation group and the control group, 30 cases in each group.children in the observation group were treated with sodium phosphate creatine, and the control group was treated with 2-linolenic acid,the clinical outcomes, clinical signs, BNF, FABP, LVEF and LVEDP were compared between the two groups.Results After treatment, the total effective rate of the observation group was significantly higher than that of the control group, the difference was statistically significant (P<0.05); The heart rate, respiratory rate and blood oxygen saturation of the two groups were significantly improved compared with those before treatment ( P<0.05 ) , the heart rate and respiratory rate of the observation group were lower than those of the control group, the oxygen saturation was higher than that of the control group (P<0.05).After treatment, the BNF and FABP indexes of the two groups were lower (P<0.05), and the BNF and FABP indexes in the observation group were lower than those in the control group (P<0.05); The LVEF of the observation group was higher than that of the control group, the LVEDP index was lower than that of the control group, the difference was statistically significant (P<0.05).Conclusion Bairui granule and Deacetylated Ligustrazin injection in the treatment of heart failure with pneumonia has a good effect in the process, which will help reduce the BNP index in children, recovery of children's body function.

Objective To study the efficacy and safety of CO2 laser microsurgery for early glottis carcinoma.Methods The data of 96 cases of early glottic laryngeal carcinoma treated with CO2 laser microsurgery were retrospectively analyzed.Surgical specimens were conventionally embedded with paraffin and serially sectioned.Sections were stained by using hematoxylin-eosin for pathologic examination.The local control rate was observed after operation.Results Among the 96 specimens,88 surgical margins were negative and 8 were positive.10 patients (10.4 ％) recurrenced followed up for 3 years.The recurrence rates of Tis,T1a,T1b and T2 were 0 (0/10),4.1％ (2/48),25.0 ％ (4/16) and 18.1％ (4/22),respectively,with significant differences among groups (X2 =6.105,P ＜ 0.05).All of 8 cases with positive margin and 2 cases with negative margin were recurrened.The recurrent rate of patients with involvement of the anterior commissure was 30.0 ％ (6/20),which was higher than that of patients with no tumor invasion [5.3 ％ (4/76)] (X2 =9.624,P ＜ 0.01).Conclusions The safe edges can be obtained by CO2 laser microsurgery for early stage of glottis carcinoma,which have advantages in local control rate,vocal function of the larynx and curative effect.