Objective To examine the expression of metastasis-associated in colon cancer 1 (MACC1) gene in ovarian cancer cell lines and investigate its effect on biological behaviors of ovarian cancer cells.Methods The expression of MACC1 was examined by qRT-PCR and Western blot analysis in four ovarian cancer cell lines inculding OVCAR3,ES-2,SKOV3 and HO-8910.When the MACC1 was transfected to OVCAR3 cells,fluorogenic quantitative PCR was used to filter and identify MACC1 gene after the efficient silencing.Changes of adhesion in the cells were analyzed by an adhesion assay.Transwell migration and invasion assay and in vitro vascular mimicry assay were used to detect migration,invasion and angiogenesis of OVCAR3 cells in vitro.Results The expression of MACC1 gene was higher in OVCAR3 compared to other cell lines.qRT-PCR confirmed that the expression of MACC1 was silenced successfully after transient transfected MACC1-siRNA into OVCAR3 cells.After successful silencing the MACC1 expression,the adhesion ability was inhibited to some degree.In transwell migration assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than control groups (245.5 ±12.8,500.3±16.5 and 496.3±13.1 respectively),while in transwell invasion assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than the negative group and control group (185.3±14.1,405.7±9.1 and 416.3±11.5 respectively),both with markedly differences among the three groups.In tube formation assay,the distrubition of HUVECs was diffused with less junctions,and the average number of complete tubular structure was decreased in transfected group compared to the corresponding controls.Conclusion RNA interference inhibits the expression of MACC1 and effectively inhibits the metastasis and invasion abilities of ovarian cancer cells in vitro,and MACC1 is expected to become the target gene of ovarian cancer treatment.

Objective To investigate the inhibitory effects of RNA interference (RNAi) on the expression of matrix metalloproteinase-2 (MMP-2) gene and growth, adhesion,invasiveness and migration of ovarian cancer cells. Methods One specific target sequence of MMP-2 gone and one non-specific sequence (NC group) were chosen,the medium DMEM as blank group.After transfection of ovarian cancer OVCAR-3 cells, the RT-PCR and Western blot were used to detect mRNA and protein expression of MMP-2 gene, the growth ability was detected by MTT assay, the abilities of adhesion was detected by cell adhesion assay, the invasion and migration were detected by Matrigel invasion assay and wound healing assay. Results By contrast to the NC group,the mRNA expression was decreased by 73.8 ％,78.8 ％ and 78.4 ％(P＜ 0.05) in 24 h,48 h and 72 h after transfection and protein expression was decreased by 72.6 ％,81.2 ％ and 76.4 ％(P＜ 0.05) respectively at the same time. The 48 h group had the most efficient inhibitory effect. Cell growth curve revealed that cell growth was not significantly inhibited (P＞ 0.05). Adhesion was significantly reduced,the inhibitory rate was 55.0 ％ at 60 min and 44.8 ％ at 90 min (P＜ 0.05),respectively. Invasion and migration were significantly reduced as well,the inhibitory rate on invasion and migration were 29.7 ％ and 35.8 ％(P＜0.05), respectively. Conclusion siRNA mediated MMP-2 down-regulation in ovarian OVCAR-3 cells can inhibits its adhesion,invasion and migration,but do not significantly affect its growth,suggesting a important target to ovarian cancer gene-therapies.

A pilot-scale research on purification of odorous gas emitted from wastewater treatment plant using a biofilter was conducted. The aim of this study is to check on the performance of biofilter running in various conditions and the effect of pH fluctuations on the performance of biofilter. The relation between distribution of microorganism and removal of odorous gases were also discussed here. The experimental results show that the predominant odor-causing gas can be efficiently eliminated by a biofilter inoculated with deodoring microorganism which were isolated previously. Moreover the biofilter had been proved having good tolerance to shocking loads of pollutant and can operate well in the condition of low pH.

Objective To explore the effects of endoplasmic reticulum stress response(ERS) and its related apoptosis on dopaminergic neurons death.Methods NGF treated-PC12 cells were treated with 6-OHDA,MPP+ and rotenone.MTT assay and flow cytometry were used to measure the cell viability and the rate of cells apoptosis induced by those neurotoxins at different concentrations and times.The expressions of ERS-related gene XBP1,Grp78,CHOP,caspase-12 in drug-treated group and reserpine preincubation group were determined by RT-polymerase chain reaction(RT-PCR) and immunohistochemistry.Results After exposing to different concentration toxins,the vitality of PC12 cells was decreased by 52% at 100?mol/L 6-OHDA,by 44% at 75?mol/L MPP~+,and by 40% at 20 nmol/L rotenone for 24 hours respectively and ws decreased in a dose dependent manner. FCM assay confirmed time-dependent cell apoptosis.The apoptotic cells ratio of 24 h groups were (31.22?3.21)%,(27.46?2.35)%,(29.26?2.53)%,respectively(P＜0.01).In 6-OHDA groups,the gene expressions of XBP1,Grp78 were approximately 2-fold increased after 8 h exposure, CHOP reached peak level at 16 h(149.5?3.3% vs 35.9?1.8%,P＜0.01).The transcription level of caspase-12 was significantly higher than normal control at 16h[(95.4?2.8% vs(23.8?3.0)%, P＜0.01],but was alleviated by reserpine prcincubation(62.15?4.3%,P＜0.05).The increased expressions of Grp78 and CHOP after drug exposure were confirmed by immunochemistry stain.The similar results were observed in MPP~+ and rotenone groups.Conclusions The excessive ERS and ERS-activated cell apoptosis pathway may be involved in selective death of dopaminergic neurons.

Objective To study the method of internal boundary parameters identification of middle ear.Method The numerical model is created using CT technology.Based on Matlab tools,the neural network for identifying internal boundary is proposed.Result The uniform pressure of 105 dB is applied at the outside of the tympanic membrane,and the harmonic analysis is calculated on the model to take the training samples.The internal condition parameters are identified using the good neural network.Conclusions The investiga-tion shows that the inverse method reveals a fast convergence and a high degree of accuracy.

Objective To study the method of internal boundary parameters identification of middle ear.Method The numerical model is created using CT technology.Based on Matlab tools,the neural network for identifying internal boundary is proposed.Result The uniform pressure of 105 dB is applied at the outside of the tympanic membrane,and the harmonic analysis is calculated on the model to take the training samples.The internal condition parameters are identified using the good neural network.Conclusions The investiga-tion shows that the inverse method reveals a fast convergence and a high degree of accuracy.

OBJECTIVETo detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.

METHODSThe expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.

RESULTSThe positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.

CONCLUSIONhTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Telomerase ; analysis ; genetics

OBJECTIVE To investigate the protective effect and mechanisms of luteolin-7-O-β-d-glucuronide (LGU) on oxygen glucose deprivation (OGD)-induced H9C2 cardiomyocytes injury. METH-ODS The protective effect of LGU on OGD-induced H9C2 cardiomyocytes death were investigated by MTT assay. The microfilament change of H9C2 cardiomyocytes was detected by phalloidin staining and the lactate dehydrogenase (LDH) leakage rate was also detected by LDH kit. In order to explore the possible mechanisms of LGU, ATP content, intracellular Ca2+fluorescent intensity and concentra-tion, mitochondrial membrane potential (MMP)and the expressions of apoptosis-related proteins were detected by ATP kit,CLSM(Fluo-3/AM probe),Ca2+kit,CLSM(JC-1 probe)and western blotting meth-od, respectively. RESULTS The inhibition of H9C2 cardiomyocyte survival rate inducedby OGD was improvedby pretreated with LGU in a concentrationdependent manner. The microfilaments injury as well as the increase of LDH leakage rate were also improvedby pretreated with LGU.The ATP content was significantly decreased,intracellular Ca2+fluorescent intensity and concentration were significantly increased and the MMP was significantly decreased 4 hafter OGD. LGU significantly reversed the de-crease of intracellular ATP content,the increase of Ca2+fluorescent intensity and concentration and the decrease of MMP.The release of cytochrome C,the expressionsof caspase-9 and caspase-3 in H9C2 cardiomyocytes were increased 16 h after OGD.LGUsignificantly inhibited the changes of these apop-tosis-related proteins. CONCLUSION LGU has a significant protective effect against OGD-induced H9C2 cardiomyocytes injury through inhibiting calcium overload,increasing ATP content,improving mi-tochondrial function and inhibiting apoptosis.

OBJECTIVETo investigate the role of inhibitor of DNA binding-1 (Id-1) gene in adenoid cystic carcinoma cell growth and invasion behavior.

METHODSWith salivary adenoid cystic carcinoma cell lines ACC-M and ACC-2, dedected Id-1 gene expression was screened with immunofluorescence assay. After Id-1 mRNA knocking-down using small interfering RNA, RT-PCR and Western blot were used to detect the different expressions before and after interference, and the growth of cells before and after interference was deceted using the MTT assay, and the cell invasion ability was checked with the use of Transwell chamber assay.

RESULTSId-1 were both expressed in the ACC-M and ACC-2, and the expression in ACC-M was higher than that in ACC-2. After Id-1 RNA interference, the growth and invasiveness of ACC-M and ACC-2 were inhibited with the restrained degree in ACC-M much stronger than that in the ACC-2.

CONCLUSIONIn view of the important role of Id-1 in the behavior of growth and invasion in ACC cell, interfering the expression of Id-1 gene is expected to be a novel and effective means for the treatment of adenoid cystic carcinoma.

Carcinoma, Adenoid Cystic ; Cell Line, Tumor ; Cell Proliferation ; DNA ; DNA-Binding Proteins ; Gene Silencing ; Humans ; RNA, Messenger ; Salivary Gland Neoplasms

Objective To explore the mechanism of deep brain stimulation(DBS)therapy to Parkinson's disease(PD).Methods We produced hemi-parkinsonian rat model with stereotaxically injecting 6-OHDA to right medial forebrain bundle(MFB)and stimulated ipsilateral subthalamu nucleus (STN)with platinum electrodes chronically to investigate the influence of DBS to the expression of Calbindin-28,synaptophysin and tyrosine dioxydase(TH)in Striatum by Western blot.In addition,slices of bilateral PD rats after DBS were stained to observe the expression of Calbindin-28 and synaptophysin in substantia nigra by Immunohistochemistry.Results High frequency stimulation impaired the rotational frequency 31% of unilateral PD rats triggered by apomophine;Long-term DBS increased the expression of TH in innocent striatum of unilateral PD rats 78.6%?9.5%,since the ipsilateral striatum(lesion side) was TH depleted by 6-OHDA insults;Calbindin-28 expression in ipsilateral striatum of hemi-PD rats raised up 75.4%?15.0% and long-term DBS reduced the effect by 43.0%?7.1%,meanwhile Calbindin-28 positive neurons in substantia nigra compacta in sham,PD and DBS rats were 74.5?10.2,75.7?15.6, 33.1?7.8.However,Synaptophysin expression in substantia nigra and striatum kept stable even after long- term DBS.Conclusions Consistent to the treatment to PD patients,DBS to STN alleviated the motor disorder of PD rats,the treatment might be based on regulating the expression of Calbindin-28 and TH.