OBJECTIVE:To optimize extraction technology of saponins from Paris polyphylla. METHODS:Using paris sapo-nin Ⅰ,paris saponin Ⅱand Paridis total saponin as dependent variables,using ethanol volume fraction,extraction time and sol-vent amount as independent variables,through multiple linear regression and binomial fitting,the extraction technology was opti-mized with response surface method and predicted. RESULTS:The optimized extraction technology of saponins from P. polyphylla was as follows as 10-fold of 80% ethanol,2 times reflux extraction,100 min each time. Under the extraction technology,the ex-traction rates of paris saponinⅠwere 85.4%,82.7% and 87.1%;those of paris saponinⅡwere 85.9%,81.3% and 83.6%;and those of Paridis total saponin were 89.5%,92.1% and 90.3%(all RSD<2.0%). Measured value was 0.964 9,predicted value was 0.986 0 and deviation rate was 2.14%. CONCLUSIONS:The central composite design-response surface method is simple and reliable for the optimization of extraction technology of saponins from P. polyphylla.

AIM　To develop a sensitive and specific LC/MS/MS method for determination of amlodipine in human plasma. METHODS　Amlodipine and internal standard 4′-hydroxypropafenone were extracted from plasma using liquid-liquid extraction, then separated on a Zorbax C8 column. The mobile phase consisted of acetonitrile-water-formic acid (75∶35∶1), at a flow-rate of 0.4 mL*min-1. A Finnigan TSQ tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. Selected reaction monitoring (SRM) using the precursor → product ion combinations of m/z 409 → 238 and m/z 358 → 116 was used to quantify amlodipine and internal standard, respectively. RESULTS　The linear calibration curves were obtained in the concentration range of 0.4-16.0 ng*mL-1. The limit of quantification was 0.4 ng*mL-1. Each plasma sample was chromatographed within 3.7 min. The method was successfully used in several pharmacokinetic studies for amlodipine. More than 1 500 plasma samples were assayed within two weeks. CONCLUSION　The method is proved to be suitable for clinical investigation of amlodipine pharmacokinetics, which offers advantages of specificity, speed, and greater sensitivity over the previously reported methods.

This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis. HMrSV5 cells, a human peritoneal mesothelial cell line, were co-incubated with the supernatants of gastric cancer cells. Morphological changes of HMrSV5 cells were observed. The cell damage was quantitatively determined by MTT assay. The apoptosis of HMrSV5 cells was observed under transmission electron microscope. Acridine orange/ethidium bromide-stained condensed nuclei was detected by fluorescent microscopy and flow cytometry. The expressions of Bcl-2 and Bax was immunochemically evaluated. The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supernatants of gastric cancer cells. The supernatants could induce apoptosis of HMrSV5 cells in a time-dependent manner. The supernatants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells. These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis. The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis. Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.
Apoptosis ; Cell Line ; Cell Line, Tumor ; Coculture Techniques ; Epithelial Cells/*cytology ; Epithelium ; Peritoneal Neoplasms/pathology ; Peritoneal Neoplasms/*secondary ; Stomach Neoplasms/*pathology

Objective:To elucidate the factors influencing the differences in the survival rates of esophageal squamous cell carcinoma (ESCC) patients between the rural and urban regions in China. Methods:A total of 36,723 ESCC patients derived from the clinical data-bases containing 500,000 esophageal and gastric cardia carcinoma cases (1973-2015) of the Henan Key Laboratory for Esophageal Can-cer Research of the First Affiliated Hospital, Zhengzhou University, were analyzed. Of these patients, 33,625 were from the rural re-gions (91.6%), comprising 20,906 male patients with an average age of 58.98 ± 8.71 years and 12,719 females with an average age of 59.59 ± 8.53 years. The remaining 3,098 were from the urban regions and composed of 2,089 male patients with an average age of 60.84±9.10 years and 1,009 females with an average age of 62.46 ± 9.14 years. All the patients underwent radical esophagectomy, de-tailed histopathological diagnosis, and TNM staging. Chi square test, Kaplan-Meier, Log-rank, and Cox proportional hazards regression model were used to analyze the differences between ESCC patients from rural regions and those from urban regions and among the risk factors in prognosis. Results:Kaplan-Meier and Log-rank analysis results showed that the ESCC patients from the rural regions had significantly higher overall survival than the urban patients (χ2=12.971, P=0.000). Further analysis showed that rural patients≥50 years old and diagnosed with stage IIa and IIb (middle stage) ESCC had higher survival rates than urban patients in males and females (male:χ2=16.188, P<0.001;female:χ2=5.019, P=0.025). However, the survival rates of rural and urban patients with stage 0,Ⅰa,Ⅰb (early stage) and Ⅲa, Ⅲc, and Ⅳ (late stage) were similar (P>0.05). The results of Cox proportional hazards regression model analysis showed that age, gender, and TNM stages were independent risk factors for rural and urban ESCC patients. When the rural and urban ESCC patients were both considered, the Cox proportional hazards regression model analysis results showed that male ESCC patients≥50 years old, urban residence, and TNM stages were independent risk factors. Conclusion:Rural ESCC patients have significantly high-er overall survival than urban patients. Male, age of≥50 years old, urban residence, and TNM stages were independent risk factors for ESCC patient survival.

This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis.HMrSV5 cells,a human peritoneal mesothelial cell line,were co-incubated with the supernatants of gastric cancer cells.Morphological changes of HMrSV5 cells were observed.The cell damage was quantitatively determined by MTT assay.The apoptosis of HMrSV5 cells was observed under transmission electron microscope.Acridine orange/ethidium bromidestained condensed nuclei was detected by fluorescent microscopy and flow cytometry.The expressions of Bcl-2 and Bax was immunochemically evaluated.The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supematants of gastric cancer cells.The supernatants could induce apoptosis of HMrSV5 cells in a time-dependent manner.Th esupematants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells.These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis.The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis.Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.

Objective To characterise the alterations of serum autoantibodies for cyclinB1,p62,Koc-IMP1 and survivin in the subjects with esophgeal and gastric cardia carcinoma and precancerous lesion and their expres-sions in the esophageal and gastric cardia cancer tissue.Methods Enzyme-linked immunoassay and tumor-associated antigen mini-array (consisting of five full-length recombinant proteins,including eyefinB1-p62-Koc,IMP1 and Survivin)were applied to determine the serum level of the autoantibodies of these antigens on 376 subjects with e-sephageal and gastric cardia carcinoma and precancerous lesions.At the same time,the expression of these antigens was detected by immunohistochemical method(ABC)on 13 patients with esophageal cancer and 16 with gastric car-dia cancer.Results All of the 5 antigens determined,the linear correlation Was observed for the detection frequency of cyclinB1,IMPI and p62 in esophageal carcinogenesis,and for p62 in gastric cardia multi-stage progression from normal to precancerous and cancerous lesions(P<0.05).The detection rale with single positive antoantibody im-munoreactivity for both esophageal and gastric cardia cancers were low.However.the positive detection mte for both esophageal and gastric cardia cancer increased apparently when the multiple positive markers were combined together for analysis,which increased tO 3～5 and 3～4 folds respectively.Furthermore,the difference in autoantibody immu-noreactive rate was significant with the lesion progressed from mild tO severe precancerous lesions and to cancer both in esophageal and gastric cardia cancers(P<0.05).The positive immunoreactions of the 5 antigens were detected in cancer tissues.The positive immunostaining rates for cyclinB1,Koc,IMP1 and Survivin both in esophageal and gastric cardia cancers were higher compared to their serllin positive rate of autoantibodies I P<0.05).Of the pa-tients with positive immunostaining in the two cancer tissues,the autoantibodies in the serum for the corresponding antigens could be detected in the same patient.Conclusion The production of the tumor-associated autoantibodies is related tO antigens.The screening rate through serum tumor-associated antigen mini-array for the patients with e-sophageal and gastric cardia carcinoma and precancerous lesions has been increased apparently with combined analy-sis of multiple autoantibodies than with single one.