Vitamin C (VC) and vitamin K3 (VK3) are widely used in the fields of antioxidant,immunity improvement and hemostasis,etc.In the 1 940 s,the antitumor effect of these two vitamins was found.Research shows that the combination regimen of VC and VK3 with a ratio of 100:1 produces a maximum of synergistic anti-tumor effect.Such combination reduces the overall dose of drugs,thus avoiding the increase in their adverse effects.The combination of VC and VK3 effectively kills a variety of tumor cells,such as breast cancer,ovarian cancer,prostate cancer,bladder cancer and leukemia,etc.This combined therapy has been patent and is in phase Ⅰ and Ⅱ clinical trials.The clinical studies show that it prolongs the survival time of patients with terminal cancer,while its adverse reactions is mild.Therefore,VC in combination of VK3 is of broad application prospect.This paper reviews the synergistic anti-tumor effects of VC combined with VK3.

Objective To investigate the polymorphism of seventeen Y chromosomal short tandem repeat(STR) loci in Dong,Miao and Zhuang nationality in Guangxi,China.Then to evaluate their forensic application.MethodsDNA was extracted from oral swab of minority population with magnetic beads method and then seventeen Y-STR loci were amplified with AmpFlSTR○R YfilerTM PCR Amplification Kit System.The PCR products were detected with 3l30XL Genetic Analyzer.The allele frequencies,diversity and genetic distance were calculated.Results The information of allele distribution and gene diversity of the 17 Y-STR loci were obtained from three minority population.Total 275 haplotypes were found in three minority population.The haplotype diversity of Dong nationality is 0.9972,Miao is 0.9966,Zhuang is 0.9964.The genetic distance between Dong and Miao nationality is most close.Conclusion The haplotype diversity of 17 Y-STR loci is quite sensitive in racial identification,paternity identification and human population genetics.

Objective To explore rapamycin's inhibitory effect on proliferation of H_(22) hepatic cancer in mice. Methods In vitro study: H_(22) hepatic cancer cell lines were cultured with rapamycin, CsA, FK506, and proliferation was determined through MTT. The influences of different agents on the H_(22) hepatic cancer cell cycle were observed by flow cytometry. The vascular endothelial growth factor (VEGF) concentration of the supernatant fluid of the cultured H_(22) hepatic cancer cell was detected by ELISA. In vivo study: C57BL/6 to Balb/c mice allogenic skin transplant was established, and the H_(22) hepatic cancer cell was implanted under skin. Rapamycin, CsA, FK506 and 5-FU were fed to the mice, respectively. The effect of different immunosuppressors on the survival of skin graft was observed while the proliferation of the transplant tumor was investigated. VEGF concentration of treated mice serum was examined by ELISA. The microvessel density of the transplanted tumor was observed through immunohistochemistry staining of CD34. Results The proliferation of the H_(22) hepatic cancer cells was inhibited by rapamycin at the concentration different dose of rapamycin, the VEGF concentration of the supernatant fluid decreased significantly (P<0.05). The number of S phase cells decreased significantly compared to that of other agents (P<0.05). When rapamycin, the lengthened survival time of the skin grafts was similar to that in CsA and FK506 groups. But the tumor volume was smaller than that in CsA and FK506 groups (P<0.05). Compared to that in the control group, the VEGF concentration of mice serum decreased in rapamycin group (P<0.05), and the microvessel density of the transplant tumor was reduced greatly (P<0.05). Conclusion Rapamycin, as an immunosuppressor, significantly resists immunologic rejection and inhibits the proliferation of H_(22) hepatic cancer, thus having its advantage in treating malignant hepatic cancer with liver transplantation.

Objective:To investigate the expression profile of Ly49A in a mouse GVHD model and the ex vivo expression alteration of it in T cells exposal to CsA, IL 4, and IL 15, respectively.Methods:Based on mouse aGVHD model, two color fluorescence flow cytometry was used to assay the expression of Ly49A on splenic and bone marrow CD3 +, CD4 + and CD8 + T cell subsets, before and after engraftment in different time interval. Cell culture was used to analyse in vitro Ly49A expression on splenic T cells treated with IL 4 and IL 15.Results:①In normal B6 and F1 strain, the frequencies of Ly49A + cells in BMCs , splenocytes and PMNCs remained at a low level and almost no, if any, such cells were detected in thymus; ②The proportions of CD3 +/Ly49A + andCD8 +/Ly49A + cells rose in early stage(7th,day) , tended to decline until 5 w post engraftment, and thereafter arose and exceed that pretreatment, which was down regulated in the presence of CsA; ③With or without ConA, the CD3 +/Ly49A + proportion were up modulated in purified CD3 +splenic cells, by IL 4 and IL 15 with no different effect demonstrated between those cytokines.Conclusion:KIR +T lymphocytes were most belong to peripheral lymphocytes. There exist a immunological tolerant tendency between grafts and host in semimatched transplant, and some cytokines and immunosuppressants can enhance the tolerance.

Objective To investigate the clinical significances of CD44v6 protein expression in human gallbladder carcinoma(GBC).Methods The immunohistochemical technique was used to detect the CD44v6 protein expression in 40 cases of GBC,12 cases of chronic cholecystitis,6 cases of adenoma and 4 cases of adenomyomatous hyperplasia.Results The positive rate of CD44v6 expression in gallbladder carcinoma and benign lesions was 72.5% and 0%,respectively(P＜0.01),and it also correlated with the lymph node metastasis,pathologic differentiation and clinic staging,but there was no correlation among pathological types.Conclusion Detecting CD44v6 expression might be severed as an objective indicator for differential diagnosis,lymph node metastatic potency,tumor progress and prognosis in gallbladder carcinoma.

OBJECTIVE:To understand the public cognitive status about the adverse drug reaction(ADR),and provide refer-ence for improving the ADR reporting system. METHODS:Field to fill out and recycling questionnaire on the spot were conducted to investigate the cognition of ADR about 1 379 people from the Pearl River Delta,Eastern,Western and Northern region of Guangdong Province,and analyze related influential factors. RESULTS:Totally 1 500 questionnaires were sent out and 1 379 were effective with effective rate of 91.9%. Among the respondents,931(67.5%)could correctly answer the question of ADR concept, the correct rate showed lowest in the people lived in Northern region and with no less than 40 years old and low education;only 36.1% could correctly know the cause of ADR;only 388 (28.1%) knew the individuals can report ADR;723 (52.4%) thought hospital should be held responsible for ADR;once the ADR appeared,62.2% of respondents would choose to report to the hospi-tal. The main causes of discourage individual report of ADR were due to less know about where,how and which relevant depart-ments could report,and the report way was not convenient and the procedures were cumbersome. CONCLUSIONS:The public awareness of the ADR concept is not high and the enthusiasm of report is relatively low. Therefore,the propaganda of public aware-ness of ADR should be strengthened to encourage the public to participate in the ADR report actively,and broaden the channels of ADR personal report.

OBJECTIVE:To study influential factors for retail chain drugstore undertaking the pharmacy function of community health center based on the perspective of retail chain drugstore pharmacist. METHODS:The stratified sampling method was used to conduct questionnaire survey among pharmacists in retail chain drugstores from Pearl River Delta(Guangzhou,Foshan, Dongguan),eastern Guangdong,western Guangdong and northern Guangdong. Multivariate analysis of variance was performed to study main effects of independent variables,pairwise comparison and interaction,with social demographic characteristics(gender, age,educational level,major,profession at title,working life,location)as independent variables,using fear of workload increase,inadequate pharmaceutical care,lower wages,inadequate electronic prescription acceptance and change in the work model as dependent variable. RESULTS:A total of 242 questionnaires were sent out,involving 239 valid questionnaires with effective recovery rate of 98.8%. The results of multivariate analysis of variance showed that majors of main effect and the interaction between major and professional title affected the fear of increase in the workload;age,education and working years of main effect,and the interaction between age and location affected the fear of inadequate pharmaceutical care;professional title of main effect affected the fear of electronic prescription acceptance;age and location of main effect,and the interactions between age and professional title influenced the fear of change in work model. CONCLUSIONS:It is suggested to strengthen pharmaceutical care and electronic prescription acceptance of pharmaceutical staff in retail chain drugstore,increase the number of professional pharmaceutical staff,allocate pharmaceutical staff reasonably and consider about regional differences.

AIM: To investigate the role of mitochondrial ceramidase in mitochondrial functions, especially in the regulation of apoptosis. METHODS: pCDNA3.1/His-MtCDase plasmid, containing mitochondrial ceramidase cDNA sequence, was transfected into K562 cells by liposome, and G418 was used to screen the positive clones. A stable transfected K562 cell line was established and defined as ‘K562TC’. The differences between K562 and K562TC cells in serum withdrawal resistance and Bcl-2 protein expression were evaluated by annexin V/PI test, flow cytometry and Western blotting, respectively. RESULTS: K562TC cells with elevated Bcl-2 protein expression level identified by FCM or Western blotting showed stronger resistance to apoptosis induced by serum withdrawal than their parental cells. Inhibition of mitochondrial ceramidase expression in K562TC cells by its specific antisense oligodeoxynucleotide was correlated with a decrease in Bcl-2 protein level. N, N'-dimethylsphingosine (DMS), a sphingosine kinase inhibitor, depleted intracellular sphingosine-1-phosphate (SPP) production, also abrogated Bcl-2 protein expression in K562TC cells, while exogenous sphingosine-1-phosphate up-regulated Bcl-2 protein level in K562 cells. CONCLUSION: Mitochondrial ceramidase overexpression in K562 cells leads to markedly elevated level of Bcl-2 protein and results in more resistance to serum withdrawal. This effect is initiated not by sphingosine, the direct metabolite of mitochondrial ceramidase, but via sphingosine-1-phosphate, its phosphorylated form, indicating that mitochondrial ceramidase, through its sphingoid metabolite sphingosine-1-phosphate, up-regulates Bcl-2 protein expression in K562 cells.

Objective To investigate the clinical effects and prognostic factors of radical surgery for primary gallbladder cancer (GBC).Methods The retrospective case-control study was conducted.The clinicopathological data of 305 patients with primary GBC who underwent radical Ro resection in the First Affiliated Hospital of Xi'an Jiaotong University from 2013 to 2017 were collected,including 108 males and 197 females,aged from 30 to 88 years,with a median age of 62 years.According to the different tumor staging,patients underwent corresponding operation and adjuvant treatment based on the postoperative indication of chemotherapy.Observation indicators:(1) results of imaging and laboratory examinations;(2) treatment situations:① surgical situations,② postoperative adjuvant treatment;(3) results of postoperative pathological examination;(4) followup;(5) prognostic factors analysis.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to December 5,2018,and death was used as the end point.Measurement data with normal distribution were represented as Mean±SD.Measurement data with skewed distribution were described as M (range).Count data were represented as percentage.The survival curve and survival rate were respectively drawn and calculated using the Kaplan-Meier method.The univariate analysis and multivariate analysis were respectively done using the Log-rank test and COX regression model.Results (1) Results of imaging and laboratory examinations:results of imaging examination showed that diagnostic rates of ultrasound,CT and MRI examination were respectively 84.06％ (174/207),85.71％ (168/196) and 63.11％ (65/103).Results of laboratory examination showed that the positive rates of CA19-9,CA125 and carcinoembryonic antigen (CEA) were respectively 55.34％ (145/262),48.06％ (124/258) and 46.15％ (126/273).(2) Treatment situations:① surgical situations:305 patients underwent radical R0 resection for primary GBC,including 145 undergoing liver wedge resection + D2 lymph node dissection,61 undergoing liver wedge resection + D1 lymph node dissection,55 undergoing liver Ⅳ B and Ⅴ segmentectomy + D2 lymph node dissection,11 undergoing liver Ⅳ B and Ⅴ segrnentectomy + D1 lymph node dissection,9 undergoing right hepatectomy + D2 lymph node dissection,5 undergoing liver wedge resection + D2 lymph node dissection + partial colectomy,4 undergoing pancreaticoduodenectomy,3 undergoing simple cholecystectomy in Tis stage,3 undergoing right hepatectomy + D1 lymph node dissection,2 undergoing liver ⅣB and Ⅴ segmentectomy + D2 lymph node dissection + partial colectomy,1 undergoing liver Ⅳ B and Ⅴ segmentectomy + resection and reconstruction of portal vein + D2 lymph node dissection,1 undergoing liver ⅣB and Ⅴ segmentectomy + D2 lymph node dissection + partial resection of the stomach or duodenum,1 undergoing pancreaticoduodenectomy + resection and reconstruction of portal vein,1 undergoing right hepatectomy + pancreaticoduodenectomy,1 undergoing right hepatic lobectomy + partial gastrectomy + D2 lymph node dissection,1 undergoing right hepatic lobectomy + D1 lymph node dissection and 1 undergoing right hepatic trilobectomy + D2 lymph node dissection.Of 94 patients with unsuspected GBC,78 who were diagnosed in the other hospitals received salvage surgery in the authors' center.Twenty-one patients had postoperative surgery-related complications,including 11 with bile leakage,8 with pulmonary infection and 2 with abdominal bleeding.Two patients died in the perioperative period.② Postoperative adjuvant treatment:26 patients underwent postoperative adjuvant chemotherapy.Chemotherapy regimen:gemcitabine + oxaliplatin were used in 12 patients,gemcitabine + tegafur in 7 patients,gemcitabine + cisplatin in 6 patients,oxaliplatin + tegafur in 1 patient.(3) Results of postoperative pathological examination.The postoperative pathological type of 305 patients:257,23,6,5,4,3,3,2,1 and 1 patients were respectively confirmed as pure adenocarcinoma,adenocarcinoma combined with squamous cell carcinoma,adenocarcinoma combined with neuroendocrine carcinoma,mucinous adenocarcinoma,neuroendocrine carcinoma,adenocarcinoma combined with mucinous carcinoma,squamous cell carcinoma,sarcomatoid carcinoma,adenocarcinoma combined with sarcomatoid carcinoma,adenocarcinoma combined with signet-ring cell carcinoma.Degree of tumor differentiation:highdifferentiated,moderate-differentiated and low-differentiated tumors were detected in 37,130 and 121 patients,respectively,17 with unknown differentiated degree.Of 305 patients,16 and 32 patients had respectively vascular invasion and nerve invasion.The number of lymph node dissected of 305 patients was 8±5,with positive lymph node of 0 (range,0-9),including 121 with lymphatic metastasis (26 with jumping lymphatic metastasis).TNM staging of 305 patients:stage 0,Ⅰ,Ⅱ,ⅢA,ⅢB,ⅣA and ⅣB were detected in 7,18,13,137,57,11 and 62 patients,respectively.(4) Follow-up:245 of 305 patients were followed up for 18.0 months (range,6.0-70.0 months).The survival time,1-and 3-year survival rates were respectively 29.5 months (range,0.5-69.9 months),71.6％ and 45.8％.One hundred and twenty-two patients died during the follow-up.(5) Prognostic factors analysis:the results of univariate analysis showed that preoperative level of bilirubin,pathological type,degree of tumor differentiation,liver invasion,vascular invasion,nerve invasion,T staging,N staging and postoperative chemotherapy were factors affecting prognosis of patients with primary GBC (x2 =10.26,3.96,45.89,34.64,12.75,27.05,35.09,39.44,4.40,P＜0.05).The results of multivariate analysis showed that low-differentiated tumor,liver invasion and N2 staging were independent risk factors affecting prognosis of patients with primary GBC [odds ratio (OR)=1.90,1.71,1.46,95％ confidence interval (CI):1.34-2.70,1.15-2.52,1.17-1.82,P＜0.05],and postoperative chemotherapy was a protective factor affecting prognosis of patients with primary GBC (OR=0.35,95％ CI:0.15-0.82,P＜0.05).Conclusions For patients with primary GBC undergoing radical resection,D2 lymph node dissection should be performed routinely.The low-differentiated tumor,liver invasion and N2 staging are independent risk factors affecting prognosis of patients,and postoperative chemotherapy is a protective factor.

OBJECTIVE：To analyze th e risk factors of marketing authorization holder （MAH）pharmacovigilance outsourcing and propose risk management and control strategies ， and to provide reference for MAH to successfully implement pharmacovigilance outsourcing and regulatory authorities to formulate corresponding regulatory strategies. METHODS ：Based on the principal-agent theory and brainstorming method ，the risk factors were preliminarily determined ，and a questionnaire was designed. The questionnaire survey was carried out among pharmacovigilance staff of 200 MAH in Guangdong province by random sampling. The factor analysis method was used to statistically analyze the questionnaire data ，and the comprehensive risk factors and their respective factor scores were summarized. RESULTS & CONCLUSIONS ：A total of 200 questionnaires were distributed ， and 154 valid questionnaires were returned ，with effective recovery rate of 77.00％. The results of factor analysis method showed that there were 4 comprehensive risk factors involved in MAH pharmacovigilance outsourcing activities ，which were service providers and outsourcing management factor （3.792 score），MAH factor （3.766 score），regulations factor （3.626 score）and market factor （3.610 score）from high to low. In view of the above factors ，it is suggested that MAH should focus on improving outsourcing contract ，establishing information communication mechanism with service providers ，strengthening auditing and management，strengthening pharmacovigilance ability and personnel building ，defining appropriate pharmacovigilance outsourcing business contents ， and fully doing pre-service investigation of service providers ， when MAH conducts outsourcing of pharmacovigilance. It is suggested that relevant departments should formulate pharmacovigilance entrusted quality management specifications or guidelines as soon as possible and strengthen the supervision of service providers. It is suggested to try to establish a pharmacovigilance outsourcing industry associa tion， andstandardize the industry behavior by giving full play to the influence of the association ，so as to avoid the outsourcing risk.