Location Analysis of Volatile Oil in Angelicae Sinensis Radix by Frozen Section and Fluorescence Imaging / 中国实验方剂学杂志

ObjectiveTo establish a simple， fast and accurate method for locating the volatile oil in Angelicae Sinensis Radix based on frozen section and fluorescence imaging technology， and to reveal the distribution and accumulation of volatile oil in the roots of this herbal medicine. MethodAngelicae Sinensis Radix was used as the research material， the best frozen section conditions for the research material were established by comparing the effects of different cryoprotectants on the quality of frozen sections of Angelicae Sinensis Radix. The suitability of Sudan Ⅲ chemical staining and fluorescence localization for positioning the volatile oil were compared according to the loss of volatile oil and the complexity of operation process. ResultA new method for evaluating the quality of frozen sections of Angelicae Sinensis Radix was established. According to the evaluation equation， it was found that the highest score was obtained when the head， body and tail positions of Angelicae Sinensis Radix were treated with 20% glycerol， 15% glycerol and 20% sucrose， respectively. There was yellowish-brown oily substance in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix， which could be stained orange red or orange yellow by Sudan Ⅲ， and there was green spontaneous fluorescence in the same part under the fluorescence microscope. ConclusionThe relatively complete section of Angelicae Sinensis Radix can be obtained after being treated with cryoprotectant. The volatile oil exists in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix. This study can provide reference for observation of the accumulation sites of volatile oil in other plants.

Hypoxia-Inducible Factor 1α Regulates the Transforming Growth Factor β1/SMAD Family Member 3 Pathway to Promote Breast Cancer Progression / 한국유방암학회지

PURPOSE: The transforming growth factor β1 (TGF-β1)/SMAD family member 3 (SMAD3) pathway, and hypoxia-inducible factor 1α (HIF-1α) are two key players in various types of malignancies including breast cancer. The TGF-β1/SMAD3 pathway can interact with HIF-1α in some diseases; however, their interaction in breast cancer is still unknown. Therefore, our study aimed to investigate the interactions between the TGF-β1/SMAD3 pathway and HIF-1α in breast cancer. METHODS: Expression of HIF-1α in serum of breast cancer patients and healthy controls was detected by quantitative reverse transcription polymerase chain reaction, and the diagnostic value of HIF-1α for breast cancer was evaluated by receiver operating characteristic curve analysis. Breast cancer cell lines overexpressing SMAD3 and HIF-1α were established. Cell apoptosis and proliferation following different treatments were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and cell counting kit-8, respectively. Expression of related proteins was detected by western blot. RESULTS: Serum levels of HIF-1α were higher in breast cancer patients than in normal controls. Both SMAD3 and HIF-1α overexpression inhibited cell apoptosis and promoted cell proliferation. Treatment with inhibitors of HIF-1α and SMAD3 promoted apoptosis in breast cancer cells and inhibited their proliferation. Overexpression of HIF-1α promoted the expression of TGF-β1 and SMAD3, while SMAD3 overexpression did not significantly affect expression of HIF-1α or TGF-β1. CONCLUSION: HIF-1α serves as an upstream regulator of the TGF-β1/SMAD3 pathway and promotes the growth of breast cancer.

Current status of benign breast diseases and metabolic syndrome in professional women in Chongqing and study on relative risk factors / 中华内分泌外科杂志

ObjectiveTo study current status of benign breast diseases and metabolic syndrome in professional women in Chongqing and relative risk factors. MethodsProfessional women (2604 cases )in Chongqing were surveyed by random cluster sampling.The biochemical indicators such as blood lipid were determined by cholesterin oxidase.The indicators such as height and weight were measured by physical examination.Chi-square test and logistic regression were used in statistical analysis. Results The morbidity rate of benign breast diseases, metabolic syndrome, hyperglycaemia, hypertension, hyperlipidemia, and obesity in our study was 19.27％ (429/2226), 7.91％ ( 176/2226 ), 8.04％ ( 179/2226 ), 23.23％ ( 517/2226 ), 24.21％ ( 539/2226 ) and 20.5％ (457/2226) respectively.The difference of mobidity rate between different age group and different career had statistical significance.Office workers and civil servants were high risk population.Age was negatively correlated with benign breast diseases.There was no relation between benign breast diseases and metabolic syndrome.ConculsionsThe morbidity rate of benign breast diseases and metabolic symdrome in professional women in Chongqing is relatively high.A good lifestyle, breast self-examination and regular physicial examination are recommended.

Enhancement of Y14 and Upf1 expression in human breast cancer cell lines / 基础医学与临床

Objective To study expression enhancement and significance of Y14 and Upf1 in human breast cancer cell lines and tissue. Methods Immuocytochemistry and laser scanning confocal microscope(LSCM) were applied. Y14 and Upf1 were determined in human breast cancer cell lines(MCF-7,ZR-75-30,T47D,MDA-MB435s,MDA-MB-453, MDA-MB-231) and breast epithelial cell line ( HBL-100). Results (1) Y14 and Upf1 level of breast cancer cells are obviously higher than that in breast epithelial cell line (P < 0. 05 ). (2)Y14 and Upf1 level of MDA-MB-231 are obviously higher than that in MCF-7. (3)The expression enhancement of Y14 and Upf1 level are obviously higher in human breast cancer tissue. Conclusion The expression level of Y14 and Upf1 in breast cancer cells and tissue enhance obviously.

In vivo effect of annexin I down-regulation on the growth of human pancreatic cancer in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To further explore the effect of annexin I on the tumor growth of human pancreatic cancer in nude mice.</p><p><b>METHODS</b>To knock down the expression of annexin I in pancreatic carcinoma cells by RNAi. A nude mouse model of human pancreatic cancer was established by subcutaneous inoculation of human pancreatic cancer cell line Suit-II cells. The effect of annexin I on tumor growth was assessed by tumor growth curve and tumor weight records, and Westen blot and flow cytometry were used to examine the expression of annexin I after annexin I-knocking down.</p><p><b>RESULTS</b>The results of Western blot revealed that the expression of annexin I was significantly decreased in Suit-II cells transfected with pSilencer-annexin I-siRNA1, and almost completely inhibited in the cells transfected with pSilencer-annexin I-siRNA2 and pSilencer-annexin I-siRNA3. The growth of tumors transfected with annexin I-siRNA2 and annexin I-siRNA3 was inhibited by 76.6% and 68.4%, respectively, in comparison with that of tumor from the parent Suit-II cells. At 44 days after tumor cell inoculation, the tumor weight was 0.8987 g (transfected with annexin I-siRNA2) and 0.8992 g (transfected with annexin I-siRNA3), significantly lower (P < 0.001) than that of tumor from parent Suit-II cells (2.5866 g) and transfected with annexin I-siRNAN (2.4070 g).</p><p><b>CONCLUSION</b>annexin I promotes the growth and proliferation of pancreatic carcinoma cells in vivo and increases the ability of tumor formation in nude mice. The results of this study support that annexin I may become a potential target in gene therapy for this disease.</p>

Preparation and primary analysis of monoclonal antibodies against VP5 protein of chicken infectious bursal disease virus / 生物工程学报

Infectious bursal disease virus (IBDV), the causative agent of a highly contagious disease in chickens, carries a small nonstructural protein (NS). In this study, vvIBDV Gx-VP5 genes were cloned into plasmid pET30a( + ) and expressed in E. coli with IPTG inducing. BALB/c mice were immunized with the purified recombinant fusion protein. SP2/0 myeloma cells and spleen cells of BALB/c mice were fused by PEG(MW1500), three hybridoma cell lines were examined by indirect ELISA and clone for three times by limited dilution, and were named as 4B4, 6D12, 3E8. The subtype of the monoclonal antibodies were IgG1 with a subtype identified ELISA kit, and light chains were kappa. The ascites titers of monoclonal antibodies were 5 x 10(4), 3.5 x 10(4), 3 x 10(4) by indirect ELISA, respectively. Indirect ELISA and Western blot results showed that the monoclonal antibodies only acted with VP5 protein, IF analysis indicated that three monoclonal antibodies acted with IBDV Gt. There were specific fluorescence in detected Vero E6 cells which transient expressed VP5 protein by IFA. Therefore, monoclonal antibodies specific to IBDV VP5 proteins are specific method for detected VP5 proteins, and base on establish stabilize expressed VP5 protein Vero cell lines to research IBDV VP5 protein function.

Studies on relationship between the expression of hTNF-alpha gene and photosynthesis in Anabaena sp. IB02 / 生物工程学报

The effects of illumination on growth of Anabaena sp. IB02 and hTNF-alpha expression were studied. Photosynthetic activity, PS I and PS II activity of Anabaena sp. IB02 were assayed. Illumination enhanced the growth of Anabaena sp. IB02 and hTNF-a expression. Some relations were observed between hTNF-alpha expression and ture photosynthesis activity, PS I, PS II activity of Anabaena sp. IB02. Significant differences of the photosynthetic activity of host were detected simultaneously when hTNF-a expressed: the respiration rate increased (-68%), the light saturation point descended (+66%), all these suggested that the metabolic charge of host were increased and grow faster than wild type under low illumination.

Enhancement of Y14 and Upf1 expression in human breast cancer cell lines / 基础医学与临床

Objective To study expression enhancement and significance of Y14 and Upf1 in human breast cancer cell lines and tissue.Methods Immuocytochemistry and laser scanning confocal microscope(LSCM) were applied. Y14 and Upf1 were determined in human breast cancer cell lines(MCF-7,ZR-75-30,T47D,MDA-MB-435s,MDA-MB-453,MDA-MB-231)and breast epithelial cell line(HBL-100).Results (1)Y14 and Upf1 level of breast cancer cells are obviously higher than that in breast epithelial cell line(P

Screening and sero-immunoscreening of ovarian epithelial cancer associative antigens / 中华妇产科杂志

Objective To explore epithelial ovarian cancer(EOC)antigens that are potentially useful for cancer early detection and therapy.Methods A high quality cDNA library derived from ascites tumor cells of EOC patients(3 cases of serous EOC,1 case of mucinous EOC,and 1 case of endometrial carcinoma of ovary)was constructed,and the method of combining serological analysis of recombinant cDNA expression libraries(SEREX)and suppression subtractive hybridization(SSH)was used for screening cDNA library.All of the positive clones were sequenced and bioinformatics analysis with BLAST software in GenBank was performed.Serological mini-arrays of recombinant tumor antigens(SMARTA)was used to investigate the prevalence of autoantibodies to these antigens in both 96 ovarian cancer patients and 96 cancer-free controls.Results Fifty-five positive clones encoding different antigenic genes of EOC recognized by IgG and(or)IgM were obtained.It showed that these 55 clones derived from 45 distinct genes and these genes could be grouped into 6 classes as following according to homology with known expressed sequence tag(EST):(1)known ovarian carcinoma related genes:BARD1,et al;(2)homologous genes with other tumors:TM4SF1,et al;(3)homologous genes with special tissues:ILF3,FXR1,et al;(4) homologous genes with special function:TIZ,C1 D,et al;(5)embryo originating genes:PKHD1,et al; (6)novel genes:OV-189,et al.SMARTA results showed that the positive ratio of five EOC antigens TM4SF1(28% vs 9%),CID(21% vs 6%),BARD1(23% vs 5%),FXR1(23% vs 8%),OV-189 (31% vs 13%)which reacting with their IgG autoantibodies,three antigens TIZ(26% vs 8%),FXR1 (28% vs 11%),and OV-189(18% vs 7%)which reacting with their IgM autoantibodies in patients was higher than in controls(P

Effect of silencing hyaluronidase gene HYAL1 by RNA interference on invasive potential of human breast cancer cell lines / 中国普通外科杂志

Objective To study the effect of RNA interference (RNAi) on HYAL1 gene mRNA expression and the invasive potential of human breast cancer cell lines. Methods Chemically synthesized double stranded RNA (dsRNA) targeting HYAL1 was transfected into human breast cancer cell lines MDA-MB-231, MDA-MB453S, ZR-75 and ZR-75-30 using SiPORT Lipid. The transfection efficiency was observed under fluorescence confocal microscopy. Expression of HYAL1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Cell penetrate matrigel capacity were determined by in vitro experiment. Results HYAL1 -siRNA effectively inhibited HYAL1 mRNA expression ( P