OBJECTIVETo detect the effect of Nocardia rubra cell wall skeleton (Nr-CWS) on tumorigenicity induced by TC-1 cells and to clinically study anti-human papillomavirus effect of Nr-CWS in lower genital tract of women.

METHODSTumor model was established by injecting TC-1 cells subcutaneously in SCID mice, then divided them into 3 groups randomly and injected with isovolumetric physiological saline, 60 micrograms/ml Nr-CWS and 120 micrograms/ml Nr-CWS respectively, the growth of tumors was measured one week later. Nr-CWS was applied on 45 HPV positive women whose TCT test was normal and without cervical erosion 2-3 days after menstruation. HPV was detected again 3 months later to explore the effect of Nr-CWS on HPV infection in female lower genital tract.

RESULTSThe animal experiment showed the weight of transplanted tumors in treated group was less than that of control group (chi2=12.5, P= 0.002). The tumor inhibition rate was 59.1 percent and 84.2 percent in the groups treated with Nr-CWS 60 and 120 micrograms/ml Nr-CWS; the results of HPV detection in 23 out of the 45 cases (51.1 percent) became negative after the 3-month treatment; the viral load was reduced in 9, and there was no change in viral load in 13 cases. Significant difference was found between the rates of undetectable viral load and the natural viral disappearance rate (P less than 0.05).

CONCLUSIONNr-CWS has an inhibitory effect to TC-1 cell tumorigenesis and clinical application of Nr-CWS may eliminate the HPV infection in lower genital tract of a considerable proportion of women with HPV infection.

Adult ; Animals ; Cell Wall Skeleton ; therapeutic use ; Cervix Uteri ; virology ; DNA, Viral ; analysis ; Female ; Humans ; Mice ; Mice, SCID ; Middle Aged ; Papillomavirus Infections ; complications ; drug therapy ; Uterine Cervical Neoplasms ; drug therapy ; virology ; Viral Load

Objective To observe whether dendritic cells(DCs) transfected with recombinant adenovirus Ad5F35-LMP2 suppress the proliferation of CNE-2 in vivo. Methods Dendritic cells have been generated with the induction of IL-4,GM-CSF and TNF-a in vitro. After human PBMCs were transplant into SCID mice,SCID mice were vaccinated with Ad5F35-LMP2-DC or untransfected DC. And four days latter,CNE-2 (1×106cell for each mouse) inoculated into SCID mouse subcutaneously. The tumor growth was monitored every week. Results The tumor latent period of Ad5F35-LMP2-DC group is longer among three groups. Untransfected DC group and CNE-2 positive control group could be found tumor about one week, but Ad5F35-LMP2-DC group was about two weeks. And the tumor was smaller and lighter than untransfected DC group and CNE-2 positive control group. Average tumor control rate reached 64.75% compared with untransfected DC group. Conclusion Ad5F35-LMP2-DC could effectively suppress the proliferation of CNE2 in vivo.

Objective To investigate the effect of different concentrations of magnesium-calcium alloy extract on the expression of matrix metalloproteinase-9 (MMP9) and tissue inhibitor of metalloproteinase-3 (TIMP3) in human colonic epithelial NCM460 cells.Methods The different concentrations of extracts (the volume fraction was 10％,50％ and 100％ respectively) were made with magnesium-calcium alloy.The 5 × 106 L-1 NCM460 suspension was randomly divided into control group,experimental group 1,experimental group 2 and experimental group 3.The cells in the control group were cultured by 2 000 μL high glucose Dulbecco's modified Eagle's medium (containing 10％ volume fraction of fetal bovine serum).The cells in the experimental group 1,2 and 3 were cultured by 2 000 μL magnesium-calcium alloy extract with volume fraction of 10％,50％ and 100％ respectively.The expressions of MMP9 and TIMP3 mRNA in NCM460 cells was detected by real-time fluorescence quantitative polymerase chain reaction,and the expression of MMP9 and TIMP3 protein in NCM460 cells was detected by Western blot at after one,three and five days of cultivation respectively.Results The expression of MMP9 mRNA and TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group after one day of cultivation (P ＜ 0.05).After three and five days of cultivation,the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 was significantly lower than that in the control group (P ＜ 0.05),but the expression of MMP9 mRNA in the NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P ＜ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 3 was significantly higher than that in the experimental group 2 after five days of cultivation (P ＜ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 1,2 and 3 after three and five days of cultivation was significantly higher than that after one day of cultivation(P ＜ 0.05).There was no significant difference in the expression of MMP9 mRNA in NCM460 cells of the experimental group 1 between three and five days of cultivation (P ＞ 0.05).The expression of MMP9 mRNA in NCM460 cells of the experimental group 2 and 3 after five days of cultivation was significantly higher than that after three days of cultivation(P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the experimental group 1 after one day of euhivation (P ＜ 0.05).After three days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly lower than that in the control group (P ＜ 0.05);the expression of TIMP3 mRNA in NCM460 cells of the experimental group 2 was significantly lower than that in the experimental group 1 and 3 (P ＜ 0.05).After five days of cultivation,the expression of TIMP3 mRNA in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after three and five days of cultivation was significantly higher than that after one day of cultivation (P ＜ 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after three days of cultivation in the experimental group 1 (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after three days of cultivation was significantly lower than that after one day of cultivation (P ＜ 0.05),and the expression of TIMP3 after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 2 (P ＜ 0.05).The expression of TIMP3 mRNA in NCM460 cells after five days of cultivation was significantly higher than that after one and three days of cultivation in the experimental group 3 (P ＜ 0.05).After five days of cultivation,there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 1 and control group (P ＞ 0.05),the expression of MMP9 protein in NCM460 cells of the experimental group 2 and 3 was significantly higher than that in the control group and the experimental group 1 (P ＜ 0.05),but there was no significant difference in the expression of MMP9 protein in NCM460 cells between the experimental group 2 and 3 (P ＞ 0.05).After five days of cultivation,the expression of TIMP3 protein in NCM460 cells of the experimental group 1,2 and 3 was significantly higher than that in the control group (P ＜0.05);but there was no significant difference in the expression of TIMP3 protein in NCM460 cells among the experimental group 1,2and 3 (P ＞ 0.05).Conclusions The high concentration of magnesium-calcium alloy extract has certain influence on the expression of MMP9 and TIMP3 gene in NCM460 cells,which may lead to the early inflammatory reaction,and the mechanism may be related to the calcium ion concentration in the extract.

SELEX technology (Systematic Evolution of Ligand by Exponential Enrichment) is a new in vitro screening technology appeared and developed in the past 20 years. SELEX integrate library technology and screening techniques, screening a nucleic acid molecule from nucleic acid library by ligand-aptamer interaction. Similar to the antibodies, aptamers bind with the specific target substance. SELEX screening technology develops rapidly, and aptamer have been widely applied in biomedical field. This article briefly-overviewed the progress and its applications of SELEX technology in recent years.
Animals ; Gene Library ; Humans ; Oligonucleotides ; genetics ; SELEX Aptamer Technique ; methods

OBJECTIVETo study the effect of inhibition of 6A8 alpha-mannosidase expression on adhesiveness among and E-cadherin expression on CNE-2L2 cells, and on metastasis of the tumors from the cells inoculated in nude mice.

METHODSAnchorage-independent adhesion among cells was examined in soft agar culture. E-cadherin expression was studied by immunofluorescence staining, immunohistological staining and RT-PCR. CNE-2L2 cells were subcutaneously inoculated into nude mice. Eight weeks later tumor metastasis was demonstrated by means of histological examination of lung sections.

RESULTSCNE-2L2 cells with suppression of 6A8 alpha-mannosidase expression (AS) became aggregated. E-cadherin expression on wild type cells was very weak. In contrast, it was greatly enhanced on AS cells. The enhancement was detected on both protein and mRNA levels. Lung metastasis of the tumor from inoculated AS cells were heavily inhibited in nude mice.

CONCLUSIONInhibition of 6A8 alpha-mannosidase expression results in enhancement of cell-cell adhesion and of E-cadherin expression on CNE-2L2 cells. Lung metastasis of the tumor grown from AS cell inoculate in nude mice is heavily suppressed.

Animals ; Cadherins ; biosynthesis ; genetics ; Cloning, Molecular ; Lung Neoplasms ; prevention & control ; secondary ; Lymphatic Metastasis ; prevention & control ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; pathology ; Neoplasm Transplantation ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured ; alpha-Mannosidase ; biosynthesis ; genetics

Objective To study the cardiotoxicity of trastuzumab in human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer patients.Methods We retrospectively collected the clinical data of general characteristic,left ventricular ejection fraction (LVEF) and clinical efficacy in 102 HER2 positive metastatic breast cancer patients treated with trastuzumab and analyzed the factors which influence the changes of LVEF and the incidence of cardiotoxicity induced by trastuzumab.Results In all of the 102 patients,the complete remission and partial remission were 7 cases (6.86％) and 73 cases (71.57％) in patients firstly treated with trastuzumab-based regimens respectively.The LVEF value was (72.85 ± 4.64) ％ at baseline and decreased to (66.05 ±5.96)％ and (65.15 ±3.38)％ as the valley points at21 and 39 months in patients who continually used trastuzumab.The progression free (PFS) of the first use of trastuzumab containing regimen and trastuzumab usage time were the clinical factors influencing the LVEF decreased value (P ＜ 0.05).The cumulative incidence of trastuzumab associated cardiotoxicity (TACT) was 14.71％ (15 cases) in the whole course of LVEF monitoring.All patients had no occurrence of heart failure and trastuzumab treatment related death caused by trastuzumab.Conclusion The heart function deceased and recovered periodically under treatment with trastuzumab in metastatic breast cancer.The incidence of cardiotoxicity was stable.Long term use of trastuzumab was well tolerated and safe.

Objective To improve the existing serological early diagnosis method of nasopharyngeal carcinoma by improve the detection sensitivity. Methods The samples of 294 serum specimen from the prevention and treatment of nasopharyngeal cancer model base, involving 106 serum specimen from the patients suffering from nasopharyngeal cancer and 188 from the healthy testers. IgA/VCA antibody and IgA/EA antibody of the serums are tested by Streptavidin-biotin-antibody immunoenzymatic test and normal traditional enzyme methods, SPSS statistical software is used to analyse the test results with χ2 test and t test. Results Referring to 106 patients, the sera antibody positive rate tested by Streptavidin-biotin-antibody immunoenzymatie test method is obviously higher than that tested by traditional method;and the t test result of the GMT has significant difference in the two method. Conclusion The modified method can improve the sensitivity of serology testing, ensure the specificity of test results, at the same time, improve the detection rate of nasopharyngeal carcinoma, so it can be applied to the early screen work of nasopharyngeal carcinoma.

Objective To investigate the expression feature of peroxiredoxin Ⅲ in cervical lesions and to further understand the mechanism for cervical cancer development/progression.Methods Expression of peroxiredoxin Ⅲ was immunohistochemically detected in cervical cancer.In addition.cervical epithelia were transfected with recombinant adeno-associated virus vector containing human papillomavirus 16 E6/E7 and pemxiredoxin Ⅲ expression was detected by quantitative real time PCR and Westem blotting.Results Peroxiredoxin Ⅲ was significantly up-regulated in cervical cancer tissues.Nevertheless,expression of peroxiredoxin Ⅲ remained unchanged in cervical epitllelial cells after transfection.Conclusion It seems that Prx Ⅲ is not related to cervical cancer initiation. Up-regulation of peroxiredoxin Ⅲ in cervical cancer might be an active response to oxidative stress in malignant cells,which protects against oxidatiton-induced apoptosis.

OBJECTIVETo understand the prevalence of Epstein-Barr virus (EBV) infection in urban and rural areas of Beijing using the serological method.

METHODSTotally 589 serum samples were collected from children in Beijing urban and rural areas who were 0--14 years old and tested with Viron-Seron ELISA classic EBV virus capsid antigen IgG antibody (EBV VCA IgG) kit. Optical density of serum samples was obtained at the wavelength of 405 nanometers. Sero-positive or negative samples were determined according to standard curve and cut-off attached in ELISA classic EBV VCA IgG kits. The activity of EBV VCA IgG was calculated by using special formula. The percentage and activity of EBV VCA IgG from Beijing children were compared with SPSS 13.0 between the urban and rural areas.

RESULTSThe percentage of EBV VCA IgG seropositive samples was 83.6%, and 80.8% in those from urban and 86.2% in those from rural areas. The peak value of EBV infection was 71% seen among children under the age of 3 years, and in urban area the rate was 67.7%, which was lower than that in the rural area (75.3%), and was 82.5% by the age of 6, which was lower than the data (up to 90%) reported 30 years ago. There was a significant difference in EBV infection rate and VCA IgG activities in children at different ages between urban and rural areas (P < 0.05).

CONCLUSIONThe rate of EBV infection in children living in urban area was lower by the age of 6 years. The primary infection of EBV occurred late in part of children lived in urban area.

Adolescent ; Age Factors ; Antigens, Viral ; immunology ; Capsid Proteins ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Cities ; epidemiology ; Epstein-Barr Virus Infections ; epidemiology ; immunology ; Herpesvirus 4, Human ; immunology ; Humans ; Immunoglobulin G ; analysis ; immunology ; Infant ; Infant, Newborn ; Rural Population ; statistics & numerical data ; Serologic Tests

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of long non-coding RNA (lncRNA) in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and randomly divided into simulated microgravity (SMG) group and normal gravity (NG) group.Each group had three samples,the rotator axis of SMG group was paralleled to the ground rotation,while the rotator axis of NG group was vertical to the ground rotation,and the speed of rotation was consistent for the two groups.The samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled and hybridized in sequence.The lncRNA and mRNA were detected by Agilent Mouse lncRNA Chips respectively.Differentially expressed lncRNA were identified and then validated by RT-qPCR.GO and Pathway analysis were applied to determine the functional distribution of these target genes.The integration predictions of the lncRNA and mRNA co-expression had been proposed to refine the functional lncRNA-mRNA relationships.Results There were 238 differentially expressed lncRNAs including 134 lncRNAs up-regulated and 104 lncRNAs down-regulated,and 237 differentially expressed mRNAs including 53 mRNAs up-regulated and 184 mRNAs down-regulated significantly in mouse fibroblasts L929 cell line under simulated microgravity by RCCS.The RT-qPCR showed a high concordance with chip microarray results in 4 differentially expressed lncRNA.GO analysis showed that the differentially expressed lncRNAs were related to the biological processes such as negative regulation of megakaryocyte differentiation and negative regulation of wound healing.Pathway analysis showed that these target genes were related to the signal pathways of systemic lupus erythematosus and TGF-β.The lncRNA-mRNA co-expression networks were also established.Conclusion The simulated microgravity by RCCS could significantly affect the expression profiles of lncRNA and mRNA in mouse fibroblasts L929.The lncRNA target genes prediction and functional enrichment analysis based on gene chip technology may provide the theoretical basis for illustrating the mechanism and management of weightlessness stress injury.