Objective To investigate the efficiency of biology function of ITIH4 gene silenced by small interfering RNA (siRNA) on ovarian cancer.Methods The four pairs ITIH4 gene siRNA interference fragments(ITIH4-546,ITIH4-795,ITIH4-917 and ITIH4-1568) were designed respectively,and transfected into HO8910pm cells with ITIH4 mRNA high expression by liposomal method transiently.Quantitative PCR method was used to detect the ITIH4 mRNA expression in HO8910pm cells transfected with interference fragment.The ITIH4 917 was selected as the best silencing effect of siRNA interference fragment and then the recombinant plasmid expression vector pGPU6/GFP/Neo-shRNA-ITIH4-917 was constructed and transfected into HO8910pm cells.The stably transfected cells-pGPU6/GFP/Neo-shRNA-ITIH4-917-HO8910pm cells was obtained by screening of aminoglycoside antibiotics (G418).The experiment was divided into three groups,namely ITIH4-917 transfection group,the HO8910pm cell group transfected with pGPU6/GFP/Neo-shRNA plasmid (empty vector group),and the HO8910pm cell group transfected with pGPU6/GFP/Neo-shRNA-ITIH4-NC the plasmid (negative control group).Fluorescence quantitative reverse transcription(RT)PCR and western blot were used to detect the ITIH4 mRNA and protein expression.The cell proliferation,the cell cycle,colony formation of cells,cells migration and invasion in vitro were determined by using methyl thiazolyl tetrazolium (MTT),flow cytometry,colony formation assay and transmembrane (transwell) small chamber method [value represented by absorbance (A)],respectively.Results The fluorescent quantitative PCR results showed that the ITIH4 mRNA expression levels in ITIH4-917 HO8910pm cells was significantly lower than that in the control cells,the relative copy number was only 0.26 ± 0.15.Also the relative copy number of ITIH4 mRNA in ITIH4-917 transfection group cells was 0.34 ±0.10,it significantly lower than that in empty vector group (1.87 ±0.12,P =0.008) and negative control group (1.58 ±0.21,P =0.032) ; Western blot results showed that the ITIH4 relative expression levels of the protein in ITIH4-917 HO8910pm group cells,empty vector group and negative control group were 0.51,1.64 and 1.74,respectively,there were statistically significant differences (0.51 vs.1.64,P =0.012; 0.51 vs.1.74,P =0.014).MTT colorimetric assay showed that the proliferation of ITIH4-917 HO8910pm group cells was significantly faster than that in the empty vector group and negative control group,and there were statistically significant differences among them (P =0.001).The S ± G2/M phase cell ratio in ITIH4-917 HO8910pm group cells was 54.2％,which was significantly higher than that in the empty vector group or negative control group (26.3％ and 31.3％,respectively,all P ＜ 0.05).The colony formation rate (55.7 ± O.7) ％ in ITIH4-917 HO8910pm group cells was also significantly higher than that in empty vector group (29.7 ±0.9) ％ (P =0.037) and negative control group (31.4 ± 0.3) ％ (P =0.043).Migration and invasion experiments showed that cell migration in ITIH4-917 HO8910pm group cells was 0.40 ± 0.18,whicht was significantly higher than that in the negative control group or empty vector (0.30 ±0.03,P =0.031 ;0.25 ±0.03,P =0.028,respectively).Although the invasive ability of ITIH4-917 HO8910pm group cells (1.31 ±0.34) was higher than that in the control cells (1.05 ±0.68) and empty vector group (1.14 ±0.08),while there were not significant difference (P ＞ 0.05).Conclusion It would be to promote the cell doubling time and increase the migration capability in HO8910pm cells that ITIH4 expression was down-regulating by ITIH4 mRNA interference.

Aim Telomerase is highly expression in most tumor cells, and it is an ideal target for cancer molecular targeting therapy. It has been proved that wogonin effectively inhibits telomerase activity and tumor cell growth in vitro. The study was to explore the inhibitory effect of wogonin on the growth of tumor and telomerase activity of implanted human ovarian cancer cell line SKOV3 in nude mice. Methods Nude mice with implanted human ovarian cancer cells SKOV3 were randomly divided into five groups, viz. the high dose group of Wogonin(600 mg?kg-1),low dose group of Wogonin(300 mg?kg-1),normal control group, cisplatin therapy group(3 mg?kg-1), and combined therapy group(cisplatin plus wogonin).The weight of nude mice and the volume of tumor were regularly measured. DNA、RNA and protein were extracted from the tumor tissue. The length of telomere was examined by Southern blot. The expression of telomerase hTERT gene was detected by RT-PCR. The telomerase activity was examined by TRAP-PCR-silver staining. Results The wogonin significantly inhibit the growth of tumor when compared with controlled group.The inhibitory rate of high dose group and low dose group were 56.67% (P=0.002) and 38.10%(P=0.019), respectively. The inhibition rate of cisplatin therapy group was 50.83%(P=0.004). The suppress rate of combined group reached 66.9% and higher than any single therapy(P=0.002). The length of telomere in different concentration groups of wogonin was the same as that in the control group.Wogonin inhibited the expression of telomerase gene hTERT and telomerase activity. The inhibition is related to the dose of wogonin. Conclusion Wogonin suppresses the growth and telomerase activity of tumor. The inhibitory effect is related to the dose of wogonin. Combination of wogonin and cisplatin increase the inhibitory rate in nude mice tumor.

Objective To discover radioresistance associated molecular biomarkers and its mechanism in nasopharyngeal carcinoma by protein-protein interaction network analysis.Methods Whole genome expression microarray was applied to screen out differentially expressed genes in two cell lines CNE- 2R and CNE-2 with different radiosensitivity.Four differentially expressed genes were randomly selected for further verification by the semi-quantitative RT-PCR analysis with self-designed primers. The common differentially expressed genes from two experiments were analyzed with the SNOW online database in order to find out the central node related to the biomarkers of nasopharyngeal carcinoma radioresistance. The expression of STAT1 in CNE-2R and CNE-2 cells was measured by Western blot.Results Compared with CNE-2 cells,374 genes in CNE-2R cells were differentially expressed while 197 genes showed significant differences.Four randomly selected differentially expressed genes were verified by RT-PCR and had same change trend in consistent with the results of chip assay. Analysis with the SNOW database demonstrated that those 197 genes could form a complicated interaction network where STAT1 and JUN might be two key nodes.Indeed,the STAT1-α expression in CNE-2R was higher than that in CNE-2 (t =4.96,P ＜ 0.05).Conclusions The key nodes of STAT1 and JUN may be the molecular biomarkers leading to radioresistance in nasopharyngeal carcinoma,and STAT1-α might have close relationship with radioresistance.

Objective A state quo survey of laboratory animal resource in Guangdong province is performed to provide reference data for government management decision-making and market assessment of laboratory animals. Methods We used questionnaires focusing on the laboratory animal facilities with authorization by Guangdong Province Government, which mainly included the production and use of laboratory animals,the qualification of employees and the facilities space. Results The total production and use of laboratory animals(except for the eggs)had been increasing in the last four years. 1.57 million laboratory animals were produced and 0.754 million laboratory animals were used in 2016. There were 2352 employees,roughly the same as in 2015. The facilities space for breeding was 121008 m2,and for animal experiment was 73470 m2,which were rising in the past three years. Conclusions In order to reinforce the industry development of laboratory animals in Guangdong province,some suggestions were given in our study,such as facilitating the application of superiority resource including non-human primate and aquatic laboratory animals,supporting the standardization production of several scarce mice and rats, improving relevant employees' overall level and constructing laboratory animal facilities sharing platform.

Objective:To investigate epidemiological characteristics of arsenic poisoning-related skin lesions in an arsenic tailing area in Hunan Province.Methods:A cross-sectional study was conducted. From October 2016 to January 2017, all residents aged over 18 years (except pregnant women) were enrolled from 3 villages in Baiyun Town, Shimen County, Hunan Province by using a cluster-sampling method. Demographic information was collected through a face-to-face questionnaire interview. All residents received skin examination performed by professional dermatologists, and blood, urine, and hair samples were collected for the measurement of arsenic levels. Non-conditional logistic regression analysis was performed to analyze factors associated with arsenic poisoning-related skin lesions.Results:A total of 1 092 eligible residents in the arsenic tailing area were recruited in this study, and 756 (69.2%, 95% CI: 66.5%, 72.0%) presented with arsenic poisoning-related skin lesions, including hyperkeratosis, hypo- or hyper-pigmentation. The median ( Q1, Q3) arsenic levels were 0.31 (0.14, 0.74) μg/g in hair samples ( n = 1 079), 0.84 (0.67, 1.10) μg/L in blood samples ( n =1 091), and 60.31 (41.71, 91.52) μg/L in urine samples ( n =1 092). Multivariable analysis showed that the occurrence of arsenic poisoning-related skin lesions was associated with age, residential location, and occupational arsenic exposure history, but was not associated with gender, ethnicity, education levels, migration history, arsenic levels in hair, blood, or urine. Compared with the group aged 18 - 39 years, the group aged 40 - 59 years and the group aged over 60 years showed significantly higher risks of arsenic poisoning-related skin lesions (adjusted OR = 11.34, 95% CI: 5.98, 21.50, P < 0.001; adjusted OR = 71.82, 95% CI: 35.81, 144.05, P < 0.001, respectively). Compared with the residents in the Wangyangqiao village, residents in the Heshan village and Huangchang village showed significantly higher risks of arsenic poisoning-related skin lesions (adjusted OR = 2.89, 95% CI: 2.05, 4.08, P < 0.001; adjusted OR = 4.13, 95% CI: 1.94, 8.78, P < 0.001, respectively). The risk of arsenic poisoning-related skin lesions was significantly higher in residents with occupational exposure history than in those without (adjusted OR = 1.99, 95% CI: 1.04, 3.83, P = 0.039) . Conclusion:Nearly 70% of the residents presented with arsenic poisoning-related skin lesions in an arsenic tailing area in Hunan Province, and the duration and previous degree of arsenic exposure were associated with the risk of arsenic poisoning-related skin lesions.

【Objective】 To study the status and conduct effect evaluation of blood donation recruitment of blood services in Chongqing, and explore its influencing factors, so as to provide reference for the regional homogenization of blood services in Chongqing. 【Methods】 19 blood services in Chongqing were investigated by questionnaire in terms of the input in human resources and funds, recruitment methods, document construction and effect evaluation. The statistical analysis was conducted. 【Results】 The average number of blood donors per 1 000 population in 19 blood services in Chongqing was 9.35±3.35. Among the 19 blood services, blood inventory warning occurred in 18, 6 of them reached Level 2 and 1 of them was Level 1. The number of blood donations per 1 000 population in blood banks with no more than 5 recruits or with less than 100 000 yuan/year recruitment fund was significantly lower than that in blood banks with more than 5 recruits or with more than 100 000 yuan/year recruitment fund(P<0.05). SMS and telephone recruitment were most commonly used in blood donation recruitment. Most blood banks have established corresponding system documents, but only one has established the method to evaluate the effect of blood donation recruitment. 【Conclusion】 The number of blood donations per 1 000 population in 19 blood services in Chongqing varies greatly, and the pressure of blood inventory warning is widespread. The input of human resources and financial fund have a certain impact on the number of blood donations per 1000 population, but not the alone factor. The recruitment method is a little bit more on the traditional side, and the blood donation recruitment and efficacy evaluation is in lack of documentary supporting. Regional homogenization should be achieved by integrating the resources of blood services, establishing the document framework of blood donation recruitment and effect evaluation, clarifying the evaluation content and unifying the evaluation standard.

OBJECTIVE: To establish a method for directional screening of the cytotoxic components from the medicinal herb of Achnatherum inebrians by a combination of surface plasmon resonance (SPR) biosensor and chromatographic isolation technology. METHODS: Under the guidance of bioactive assessment based on binding abilities between objects and the α-Mannosidase (α-Man) target, the active components from different solvents extracts, different polar extraction parts and fractions were screened orderly and directionally using SPR. Components with a high binding ability to α-Man can be precisely oriented in a narrower fractions range and are easy to isolate. Three human cancer cells were used to evaluate the cytotoxic activity of component with the highest affinity to α-Man. RESULTS: Eight compounds were isolated and identificated from A. inebrians for the first time. Deoxyvasicinone possessed the highest affinity to α-Man among them. Moreover, deoxyvasicinone showed good effects on inhibited proliferation of human hepatoma cells HepG2 (IC₅₀ = 5.7 μmol/L), human breast cancer cells MCF7 (IC₅₀ = 7.21 μmol/L) and human lung cancer cells HCC827 (IC₅₀ = 0.75 μmol/L), respectively. In particular, its inhibitory effect on HCC827 was stronger than the positive drug gefitinib (IC₅₀ = 1.65 μmol/L). CONCLUSION A comprehensive strategy of directional screening potential cytotoxic components from herb based on biomolecular interaction and chromatography was established. Deoxyvasicinone as an effective anti-cancer component was initially isolated from A. inebrians. It is expected that this screening strategy could provide new perspectives for rapid screening and identification of active components from natural plants with the complex matrix.