Purpose:To explore the role of terminal restric tion fragments (TRFs), telomerase activity and expression of human telomerase re verse transcriptase (hTERT) in colorectal carcinoma. Methods:The telomere length, telomerase activity and expression of hTERT were studied with Southern-blot, TRAP and immunohistochemistry, respe ctively. Results:TRFs in cancer tissue was much shorter than in the adja cent tissues and normal mucosa, and TRFs was decreased gradually along with the development in cancer stage.The expression of Telomerase in colorectal carcinoma tissue was significantly higher than that in other tissues (P

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 clone cathepsin L (CTSL) gene and construct the eukaryotic expression plasmid pcDNA3. 1-CTSL and study the relationship between CTSL and invasion and metastasis in ovarian cancer cells in vitro. Methods The total RNA was extracted from the ovarian cancer tissue and the intact cDNA of CTSL was applied by reverse transcription (RT)-PCR. The product of RT-PCR was cloned to pMD18-T vector, and subcloned to pcDNA3. 1 vector. It was tested by the enzymation and DNA sequencing.The eukaryotic expression plasmid of CTSL was introduced into HO8910 cells by liposome transfection reagent. RT-PCR was used to confirm the recombinant plasmid DNA integrated with the genomic DNA of HO8910 cells. Western blot was used to confirm the CTSL protein expression in positive clones cells. The cell growth curves, clonogenicity efficiency were observed. The cell cycles were measured by flow cytometer.The ability of invasion, metastasis and adhesion of ovarian cancer cells were detected by the matrigel invasion assay, transwell migration assay and adhesion assay, respectively. Results The results from restrictive enzyme analysis and sequencing showed that the CTSL gene was successfully inserted into pcDNA3. 1.Result from RT-PCR and western blot showed that the ovarian cancer cells which transfected by recombinant plasmid could express CTSL gene and protein. There was no difference between HO8910-CTSL and HO8910-pcDNA3. 1 cells in proliferation and adhesion ability (0.16±0.04 versus 0. 19±0. 04) of the cells (P>0.05). There was difference between HO8910-CTSL and HO8910-peDNA3.1 cells in matrigel invasion ability (0.34±0.18 versus 0.17±0.04) and metastasis ability (1.252±0.114 versus 0.486±0.027) of cancer(all P<0.05). Conclusion CTSL maybe increase the ability of invasion and metastasis of ovarian cancer cells in vitro, which may be a molecular target of blocking invasion and metastasis of ovarian cancer.

Objective To assess the suppression effect on WWOX gene in SKOV3/SB cell line by small interference RNA (siRNA). Methods Transfection of siRNA using lipofeetamine 2000 was conducted to silence WWOX gene expression, the expression levels of WWOX mRNA and protein were evaluated,and the effects on the cell cycles at 48 hours of transfection were assessed by RT-PCR, western blot and flow eytometry (FCM) respectively. The cisplatin resistance index was assayed after transfection of SKOV3/SB with siRNA by methyl thiazolyl tetrazolium(MTT) and the cisplatin concentration of SKOV3/SB cells transfected with siRNA of WWOX was measured by high performance liquid chromatography. Results(1) In SKOV3/SB cells transfected with WWOX interference fragment, whether at the mRNA or protein level, the expression of both of WWOX decreased. There was a significant difference (P<0.05) compared with SKOV3 cells and non-transfected cells. (2) After transfecfion of the WWOX interference fragment, the index of platinum resistance of SKOV3/SB decreased from 5.04 to 3.89. (3) The number of cell transfected with the WWOX interference fragment in G1 phase was increased, while that in S-phase was decreased. (4) The cisplatin concenla'ation of SKOV3/SB cells transfected with the WWOX interference fragment was increased from 9.43 ng/L to 23.45 ng/L compared with SKOV3/SB cells non-transfected with a significant difference (P<0.05). Conclusion WWOX gene may be involved in cisplatin resistance phenomenon in epithelial ovarian cancer.

Objective To detect the genetic transcription and protein expression level of endoplasmic reticulum aminopeptidase 1 (ERAP1) in human ovarian cancer cells and tissue and study their relationship with directional lymphatic metastasis. Methods Real-time fluorescent quantitative PCR and western blot were used to determine the expressions of ERAP1 gene and protein in the human ovarian cancer cell lines between non-directional (SKOV3) and directional highly lymphatic metastasis (SKOV3-pm2, SKOV3-pm3, SKOV3-pm4 ). Immunohistochemistry method was used to further validate the ERAP1 expressions of the transplanted ovarian tumor primarily focus and the lesions of lymph node metastasis from nude mice and the human ovarian cancer primarily focus and the lesions of lymph node metastasis. Results The expression level of ERAP1 gene and protein were down-regulated in SKOV3, SKOV3-pm2, SKOV3-pro3, SKOV3-pm4 cell sublines (0.118±0.012, 0.031±0.003,0.028±0.003, 0.016±0.005 ; 0.91± 0.33, 0.09±0.03, 0.10±0.04, 0.05+0.04; respectively), and the level of ERAP1 in SKOV3 cell was higher than those in the other three kinds of cell lines (P<0.05 ). The results showed that there were significant declining trend of expression of ERAP1 in the human ovarian cancer cell lines between non-directional and directional highly lymphatic metastasis; the transplanted ovarian tumor primarily focus and the metastasis lesions of lymph node from nude mice (143±22 vs. 97±12, P<0.05), the primarily focus (184±14) and the lesions of lymph node metastasis from human ovarian tumors (P<0.05). Conclusion The absence or down-regulated expression of ERAP1 is closely related to the metastasis and invasion of lymph node in ovarian carcinoma.

AIM To observe the effects of Oxymatrine on induction of apoptosis in human ovarian cancer SKOV3 cells. METHODS Apoptosis induced by Oxymatrine in human ovarian cancer SKOV3 cells was tested by MTT assay, fluorescent microscope, and DNA gel eletrophoresis. RESULTS SKOV3 cell viability dropped down depending on the Oxymatrine concentration and treatment time. When incubated with Oxymatrine (0 189 mmol?L -1 and 0 378 mmol?L -1 ) for 48 h, SKOV3 cells showed morphological changes associated with the characters of apoptosis under fluorescent microscope. Typical DNA ladder was found during gel eletrophoresis. CONCLUSION Oxymatrine can induce apoptosis in SKOV3 cell lines.

Since the outbreak of COVID-19 caused by the 2019-nCoV (SARS-CoV-2), with its high pathogenicity and contagiousness, it has posed a serious threat to global public health security. Up to now, the pathogenesis of 2019-nCoV is unclear, and there is no effective treatment. Vaccine as one of the most effective strategies to prevent virus infection has become a hot area. Based on the current understanding of 2019-nCoV, the development of 2019-nCoV vaccines covers all types: inactivated virus vaccine, recombinant protein vaccine, viral vector-based vaccine, mRNA vaccine, and DNA vaccine, etc. In this review, we focus on the candidate targets of the novel coronavirus, and the types, development status and progress of 2019-nCoV vaccines in order to provide information for further research and prevention.

AIM To investigate the effects of Tea Po lyp henols on induction of apoptosis in human liver cancer cell etc. Telomerase acti vity of BEL-7404 cells was dramatically declined during the cell incuba tion with Tea Polyphenols. METHODS MTT colorimetric assay and trypan b lue exclusion method were used to examine the growth inhibition ofTea Polyphenols on BEL-7404 cells. Telomerase activity in cell was determined by PCR-ELISA method. Cell morphology and DNA gel electrophoresis were used to observe the apoptosis. RESULTS The solution of Tea Polyphenols showed dose-dependent and time-dependent inh ibitory effects on the proliferation on BEL-7404 cell line. After treatment wit h 0 2 and 0 1 g?L -1 tea polyphenols for 48 hours, cells displayed DNA ladder bands and typical morphological change of apoptosis included: ce ll shrinkage, membrane blebbing, chromatin condensation, nuclear fragmentation etc. Telomerase activit y of BEL-7404 cells was dramatically declined during the cell incubation with Tea Polyphenols. CONCLUSION Tea Polyphenols has growth inhibiti ng effect and may induce apoptosis in BEL-7404 in a certain dose range. The ant icancer mechanism of Tea Polyphenols might be related to the inhibition of telom erase activity.

Aim To investigate the effects of emodin isolated from Guangxi P.multiflorum Thunb on the expression of KU70/KU80 in hypoxic nasopharyngeal cancer CNE-1 cells and reveal the relationship between radiosensitization of emodin monomer and DNA repair genes.Methods The expression of hypoxia inducible factor-1?(HIF-1?)and DNA double-strand break repair genes(KU70/KU80)between the experimental groups and the control group under hypoxic condition was detected by the real-time fluorescence quantitative RT-PCR.Results Expression of HIF-1? was significantly increased under hypoxia condition.HIF-1? had no change after treatment with emodin alone.The expression level of KU70/KU80 was induced by radiation.Compared with radiation alone group,radiation combined hypoxia group obviously enhanced the expression of KU70/KU80.KU70/KU80 mRNA expression significantly reduced after radiation combined with emodin under hypoxic condition.Conclusion In the hypoxic environment,emodin combined with radiotherapy can effectively inhibit the expression of HIF-1 ? and DNA double-strand break repair genes(KU70/KU80),which may be its mechanism of radiosensitization.