Objective To study the effects of different levels estradiol on inhibitory of tamoxifen on human mammary cancer cells(ER+) in vitro.Methods Estrogen receptor(ER)-positive MCF7 human breast cancer cell line was treated by the same level of tamoxifen and different levels of estradiol in vitro.Cell proliferation was evaluated by MTT assay.Results E_2 at concentrations between 1? 10~(-12) mol/L to 1? 10~(-7) mol / L significantly stimulated the growth of MCF-7.TAM(10 ?mol/ L) inhabited the growth of MCF-7 significantly.E_2 at different levels may influence inhibitory of tamoxifen on MCF-7 cell lines.E_2(1?10~(-8) mol/L) makes inhibitory of tamoxifen on MCF-7 cell lines valueless.Conclusion E_2 is the risk factor of breast cancer,and the concentration around breast cancer cells may influence the effects of TAM.

Objective To understand the effect of estradiol in different concentrations on proliferation of diverse mammary primary cells in vitro. Methods The primary cells of cancer tissue, the adjacent tissue to tumors and normal mammary tissue from patiens with breast cancer were obtained using collagenase digesting method. All the tissue samples were cultivated in vitro, and were given estradiol in different concentrations. The effect of estradiol on the proliferation of those primary cells was measured by MTT. Results Estradiol remarkedly promoted the proliferation of primary cells of cancer tissue and peritumor tissue in vitro, whose ER expression were positive. Whereas, the promotion effect of estradiol on the proliferation of normal mammary primary cells was relatively weak, and there was no correlation between the promotion effect with the expression of ER in cancer tissue. Conclusion The risks of occurrence and relapse of breast cancer would increase significantly when the concentration of estradiol is no less than 103 pmol/L in vivo.

This study examined the cytotoxicity of a new implant material modified by microarc oxidation technique. Cells on different surfaces of the implant were evaluated 2, 4 and 6 days after treatment. The results showed that cell attachment, cell morphology, and cell proliferation were influenced by the different surface treatments, and a significant increase in the osteoblast cell activity was observed on the porous MAO-Ti coating. Our results suggest that the porous MAO-Ti surface has a better biocompatibility and electrochemical performance than pure titanium surface.

In this study, in vitro chemosensitivity testing was conducted on primary cultured breast cancer cells from 96 patients with breast cancer, and the results showed that the cells from a few patients with primary breast cancer developed multidrug resistance (MDR) prior to the first chemotherapy exposure. All the cells from the recurrent cancer patients had MDR. The findings suggested that patients having MDR would benefit from high-dose chemotherapy (HDC) regimens. In vitro chemosensitivity screening, which was aimed at improving the therapeutic efficacy and minimizing side effects, helps in choosing individualized treatment for breast cancer.
Antineoplastic Agents/*pharmacology ; Breast Neoplasms/*drug therapy ; Breast Neoplasms/pathology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Screening Assays, Antitumor/*methods ; Neoplasm Recurrence, Local/*drug therapy ; Tumor Cells, Cultured

Objective To compare the image quality and scanning dose of time-subtraction and dual-energy-subtraction cerebral CT angiography, and to assess clinical application value of both methods. Methods Plain and enhanced scanning were performed on 60 patients suspected cerebral vessel diseases with dual-source CT. Dual-energy mode with tube voltages of 140 and 80 kV was used in enhanced scanning, and data of two different energy were collected in one scanning. ①Traditional removed-bone digital subtraction (time-subtraction) with plain and 80 kV enhanced scanning data were obtained. Volume render (VR) and maximum intensity projection (MIP) reconstruction were finished; ②Direct removed-bone digital subtraction (dual-energy subtraction) with 80 kV and 140 kV enhanced scanning data were obtained. VR and MIP reconstruction were finished. The image quality of VR and MIP was divided into 4 grades, and were compared as well as average effective radiological dose. All the diseases were confirmed with surgery or DSA. Average effective radiological dose was compared with time-subtraction and dual-energy subtraction. Results Internal carotid artery trunk and branch and Willis circles were displayed clearly with two methods in 60 cases. No significant difference was found (P>0.05) between total quality score of the two methods. The size, shape, neck and axis point of aneurysm in 24 cases were clearly displayed, so as the shape and extent of abnormal vessel bolus in 4 cases, while arteries and veins were also clear in artery-vein malformation; ③The average radiological dose was (26.60±0.50)mSv in time-subtraction and (22.40±0.50) mSv in dual-energy subtraction. Conclusion The normal, abnormal vessels and diseases can be clearly displayed at time-subtraction and dual-energy subtraction CTA. The effect of dual-energy-subtraction is better than that of time-subtraction CTA in no-cooperation patients, and the radiological dose is lower in dual-energy CTA.

Objective: To construct a lentiviral expression vector that inhibits enhanced green fluorescent proteins(EGFP).Methods: The EGFP shRNA was cloned into the pLL3.7 lentivirus vector digested with XbaⅠ/XhoⅠ.J558L and Hela cells were infected by lentiviruses,and the EGFP expression was observed with the fluorescence microscope.Results: The EGFP RNAi sequence was correctly cloned into the lentivirus vector with the U6 promoter,and the EGFP expressions of Hela and J558L cells were suppressed after infected by lentivirus containing EGFP shRNA. Conclusion: The lentivirus vector effectively inhibited EGFP expression,which could be used to mediate further researches on gene function.

302 teeth with wedge-shaped defect in 168 patients were restored by SDR(smart dentin replacement,n =112),glass ionomer (n =98) and light-cured composite resin(n =92) respectively.12 to 15 month follow-up showed the success rate was 96.2％,84.8％ and 86.2％ respectively(P ＜ 0.01).SDR is an ideal material in restoring wedge-shaped defect.

Hepatitis B virus (HBV) DNA replication takes place in the viral capsid that consists of 180 or 240 copies of HBV capsid (HBc or core) protein. The monomeric core protein contains an apical loop region that forms the spikes on the surface of viral capsid upon core dimerization and capsid assembly. To investigate the impact on HBV DNA replication through gene engineering at the spike of HBV capsid. plasmids expressing engineered HBc with linker-fused enhanced green fluorescent protein (EGFP) or shortened EGFP insertion at the spike region were constructed by Restriction Digestion and Ligation-independent Cloning (RLIC). The wildtype or mutant HBc construct was cotransfected with HBV1.1c(-), a plasmid containing 1.1 unit-length HBV genome with deficiency in HBc expression, into HEK293 cells, respectively. GFP signal was observed through a fluorescence microscope and HBV DNA replicative intermediates were assayed by Southern blotting to determine the expression and functions of different recombinants. Our results demonstrated that the RLIC method was effective to generate deletion or insertion in the apical loop region of HBc. Both HBc-EGFP recombinants with different linkers produced green fluorescence but with different subcellular distribution pattern. However, HBV DNA replication was not detected with the trans-complementation of these two HBc recombinants. In addition, other recombinants including the one only with the deletion of aa79-80 failed to support HBV replication. Taken together, our results suggest that RLIC is a robust method which can be broadly applied in gene engineering; different peptide linkers may have different influences on the functions of an engineered fusion protein; and HBc aa79-80 play a critical role for HBc to support HBV DNA replication.
Capsid Proteins ; genetics ; Cloning, Molecular ; Genetic Engineering ; methods ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection ; Virus Replication

Objective To know the antitumor effect and value of vinorelbine in chemotherapy with relapsed breast cancer.Methods To receive breast cancer tissue from primary and relapsed patients by surgery,then to obtain mammary cancer cells by collagenase.The antitumor effect of 5 chemotherapy drugs with breast cancer cells was measured by culture in vitro and MTT.Results With primary cancer cells from primary breast cancer,the antitumor effect of paclitaxel and docetaxel(sensitivity is 91.04%,92.54% respec- tively) is better than adriamycin,epirubicin and gemcitabine(sensitivity is 73.13%,74.63%,71.64% re- spectively;P

Objective To study the effect of ERK signalling pathway and aldose reduetase(AR)on the transforming growth factor-β1(TGF-β1)-induced expression of fibronectin (FN)in nondiabetic nephropathy. Methods Human mesangial cells(HMCs)were cultured and transfected with pCDNA3-AR and AR gene silencing with small interfering RNA(siRNA).The AR expression in the HMCs was examined by real-time PCR and Western blotting was used to detect the protein expression of AR and FN. Inhibitors of AR and ERK signalling pathway were co-cultured with HMCs, then TGF-β1 was added and Western blotting was used to analyze the protein expression of FN. Results The expression of AR, FN and p-ERK was up-regulated by TGF-β1. AR was increased by 1.8-fold, FN was increased by 1.9-fold (P＜0.05), and p-ERK was increased by 5.l-fold after stimulation with TGF-β1 (P＜0.01). HMCs transfected with AR showed stronger protein expression of FN, more than 3.6-fold in the protein level of FN was observed in HMCs (P＜0.05). The HMCs of knockdown AR gene by siRNA showed decreased expression of AR,FN and p-ERK, the level of AR mRNA in HMCs transfected with AR siRNA was 10% of the level in untransfected cells or cells transfected with control siRNA (P＜0.01). Transfection with AR siRNA attenuated TGF-β1-induced FN production, more than 70% decrease in the protein level of FN and p-ERK was observed in HMCs with AR-siRNA (P＜0.01). Conclusions AR can regulate the expression of FN with the stimulation of TGF-β1, as AR gene is one of the responsive genes of TGF-β1, which may be associated with the activation of ERK signalling pathways induced by TCF-β1.