Objective To establish a bladder cancer MDR cell line by gene transfection and to lay a foundation for the research of occurrence and reversion mechanisms of MDR. Methods The invasive bladder cancer T24 cells were transfected with mdr1 cDNA by cationic liposome (DOTAP) introduced gene transfection.The adriamycin (ADM) resistant cells were screened as MDR cells,named TADM. The MDR phenotype of TADM was identified by MTT, immunohistochemistry,immunofluorescence,flow cytometry,PCR and RT PCR. Results The relative resistant index of TADM was 41.6,mdr1 cDNA being integrated into the genome of T24.As a result,the expression of P gp and mdr1 mRNA in TADM increased. Conclusions The integration of mdr1 cDNA into the T24 cells greatly increases the expression of P gp.TADM cells manifests excellent MDR phenotype.The establishment of MDR cell lines by gene tranfection has the benefits of less time consuming,stronger drug resistivity and more stable.

Objective To analyze the CT features of hepatic artery and morphology changes of the liver in patients with cavernous transformation of the portal vein (CTPV). Methods Thirteen patients of CTPV without malignant hepatic tumor or other diseases being able to cause changes of hepatic artery such as cirrhosis and hepatophyma were collected. CT data was analyzed, and the arterial and morphology changes of the liver were observed. Results Left and (or) right hepatic arteries were found dilated in 10 patients (76.92%), and unusual flake like enhancement was found in artery phase in 3 patients (23.08%). Atrophy-hypertrophy complex (AHC) could be found only in 1 patient (7.69%). Conclusion Dilatation of hepatic artery manifests more hepatic artery blood supply, while obvious AHC is hardly found in CTPV.

Background The primary reason to trabeculectomy failure is fibrosis of conjunctiva and episclera because of progressive fibroblast proliferation and collagen deposition of the filtration bleb.Conventional methods of inhibiting bleb scarring was intraoperative application of mitomycin C (MMC),but many complications occured after surgery.Researches showed that bevacizumab was an antifibrotic agent,and whether it can suppress scarring of filtering bleb after trabeculectomy is concerned.Objective The aim of this study was to evaluate the antifibrotic efficacy of bevacizumab after trabeculectomy in rabbits.Methods Forty New Zealand rabbits were randomly divided into four groups.Trabeculectomy was performed on the right eyes of each rabbits.The rabbits received subconjunctival injection of 0.05 ml bevacizumab (25 mg/ml) at the end of operation in the bevacizumab single injection group.The same dose of bevacizumab was respectively injected at the end of operation as well as 3 days and 7 days after operation in the bevacizumab repitition injection group,and 0.05 ml normal saline solution was used in the same way in the normal saline group.In the MMC group,MMC cotton patch with 0.2 mg/ml was placed under the Tenon caplsule and scleral flap for 3 minutes during operation.The intraocular pressure (IOP),bleb area and shape were evaluated during the 28-day period.The animals were sacrificed on postoperative day 14 and 28,respectively for the histopathologic examination of bleb.The expression of CD31 in the bleb was detected by immunohistochemistry for the calculation of microvessels.All experiments were performed in accordance with the ethics code for animal experimentation and approved by the Institutional Review Board of Tianjin Eye Hospital.Results No significant difference was found in the postoperative IOP among the groups (F =0.88,P =0.47).Compared with the bevacizumab single injection group,MMC group and normal saline group,the shape of bleb was higher and much diffuse with sparse vessels 7 days after operation in the bevacizumab repitition injection group.The survival time of bleb was 27 days,19 days and 13 days in the bevacizumab repitition injection group,the bevacizumab single injection group,MMC group and normal saline group,respectively.The percentage of collagen deposition area was (49.18±1.54)％,(26.41±1.23)％,(50.68±1.87)％ and (70.63±1.81)％ at day 14 postoperative in the bevacizumab single injection group,bevacizumab repitition injection group,MMC group and normal saline group,respectively,with the largest area in the normal saline group,and percentage of collagen deposition area was significantly reduced in the bevacizumab repitition injection group compared with the bevacizumab single injection group (all at P＜0.05).The percentage of collagen deposition area was (66.82±1.53)％ at day 28 postoperative in the bevacizumab repitition injection group,while complete scarring was seen in other 3 groups.The number of microvessels was least at postoperative day 14 in the bevacizumab repitition injection group compared with the bevacizumab single injection group,MMC group and normal saline group (all at P ＜ 0.05).The number of microvessels was more in postoperative day 28 in the bevacizumab repitition injection group (3.51 ±0.31) compared with other groups (all at P ＜ 0.05).Conclusions Subconjunctival injection of bevacizumab following trabeculectomy can improve the successful rate of surgery by remaining the survival time of filtering bleb,inhibiting the bleb scarring in rabbits.

Objectives To analysis the prevalence and major risk factors of common chronic diseases among adult residents in the city zone of Shaoyang. To formulate and evaluate the measures of health intervention. Methods Using multistage stratified random sampling, 5267 residents aged 18 or above from 26 communities of Shaoyang were invited to participate in the survey, which was 1.24% of Shaoyang municipal residents. Questionnaire, physical examination, biochemistry test and B-ultrasonography on the liver and cholecystis were undertaken. Health intervention and evaluation was undertaken too. Results The prevalence of hypertension, fatty liver, cholelithiasis, high blood sugar, dystipidemia and overweight obesity was respectively 22.0% , 24.2%, 7.4%, 8.2%, 43.7%, 47.6%. The prevalence of hypertension, high blood sugar, dystipidemia and overweight obesity was down respectively significatively to 19.6%, 5.0%,35.6% and41.1% after intervention(P<0.05). Conclusions There is a high prevalence of common chronic diseases in the city zone of Shaoyang, which can be effectively reduced through health management programme.

Background Endoscopic cyclophotocoagulation(ECP) is a new approach to treat refractory glaucoma.But,the histopathological change of ciliary body after ECP has not been reported yet.ObjectivePurpose of the present study is to explore the histopathologic change of ciliary body after ECP and investigate the mechanism of lowing intraocular pressure(IOP) of ECP and compared with trans-scleral cyclophotocoagulation (TSCP).MethodECP was performed on the right eyes of 30 adult pigmented rabbits and TSCP was taken in the fellow eyes.Other two normal pigmented rabbits were as control group.The clinical examinations were performed in day 1,3,5,7,14,28,42 and 56 after operation under the slim lamp to evaluate the inflammatory reactions of conjunctiva,sclera and cornea,and the IOP was detected in the same time points.The animals were sacrificed and the eyeballs were extracted for the examination of structure of ciliary body under the light microscope in day 7,14,28,42 and 56 after surgery respectively.ResultsThe mild inflammatory response and local opacity of lens were seen under the slim lamp in the eyes after ECP,and severe inflammatory reaction was in the eyes received TSCP.No opacity of lens was found after TSCP through the observing period.The lowing-IOP extent was significantly higher in ECP group than TSCP group after operation(P＜0.01).The structure of ciliary body epithelium was normal in normal rabbit eyes.The disruption of ciliary body epithelium,especially for non-pigment epithelial layer,edema of ciliary process were seen in the early period after ECP.Fourty-two days later,the irregulation of ciliary body epithelium and atrophy of ciliary process were exhibited under the light microscope.In TSCP group,the edema,blooding and damage of ciliary process were seen,but slight inflammatory response was in the ciliary epithelium.After 42 days of surgery,atrophy of ciliary body,irregular proliferation of pigmental and non-pigmental epithelial cells,thinness of sclera and scarring of ciliary stroma were revealed.ConclusionCompared with TSCP,ECP can selectively destroy ciliary epithelium effectively and thoroughly and therefore reduce IOP.The adverse effect of ECP is the opacity of lens.Less damage of adjacent tissue is found in ECP compared with TSCP.

Objective To observe the change of intracellular rhodamine 123 distribution model in bladder cancer sensitive cells(T24) and resistant cells(TADM),and to explore the cause of the change and its relation with multidrug resistance(MDR). Methods T24 and TAMD cells were cultured together with rhodamine 123 for a certain period.The intracellular distribution and intensity of fluorescence were detected by interactive laser cytometer(ILC). Results In T24 cells,rhodamine 123 was located mainly in the perinuclear membrane area,and in TADM cells,rhodamine 123 was concentrated mainly at the two poles of nucleus.After the extracellular rhodamine 123 was washed,in T24 cells ,the intracellular fluorescent intensity decayed slowly.It took more than 40 minutes for T24 cells to eliminate intracellular rhodamine 123 completely.By contrast,in TADM cells, the intracellular fluorescent intensity decayed rapidly.It took only 15 minutes for TADM cells to eliminate intracellular rhodamine 123 completely. Conclusions (1)Intracellular rhodamine 123 was rapidly pumped out,which is the main cause of MDR in TADM;(2)The intracellular rhodamine 123 distribution model in TADM differ from that in T24,which may be another cause of its MDR phenotype.

Objective To evaluate the relieving effects of vanillin sinffing on depression-like behaviors in depressed rats and to explore the possible underlying mechanism.Methods Depression animal model established by chronic unpredictable medium intensity stress combined with isolation and destroy the olfactory bulb.The depressed rats were divided randomly into vanillin inhalation group,fluoxetine hydrochloride group,depression model group,olfactory bulbectomy with the vanillin inhalation treatment group and sham-operated group.Nervous behavioral changes had been observed at different time after the administration of 5 weeks.The concentration of brain derived neurotrophic factor(BDNF) in the brain homogenate and the positive expression of BDNF in hippocampus had been also measured.Results Two weeks after the intervention,the immobility time of vanillin group((12.78 ±7.50) s) was lower than that of the model group((57.33±32.16) s) (P＜0.05).The consumption of saccharose in vanillin group((52.88±25.18)g) was higher than that of model group((37.40±19.33) g) (P＜0.05).BDNF of the brain homogenate in vanillin group (0.54±0.13) was significantly increased compared with model group (0.36± 0.06) (P＜0.01).When compared with the OBX group (0.40±0.06),similar result was obtained.Immunohistochemistry and the average density of image analysis revealed that the expression of BNDF of hippocampal CA3 in vanillin group (0.40±0.03)was significantly increased compared with model group (0.25±0.04) and OBX group (0.28±0.03) (P＜0.01).Conclusion Vanillin inhalation significantly relieves depression-like behaviors in depression rats.The possible mechanism may increase hippocampal neurogenesis by raising brain derived neurotrophic factor in brain.

Objective To investigate the expression changes of nuclear factor kappa B (NF-?B) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha(I?B-?)vector. Methods After pcDNA3-I?B-? vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-?B and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-I?B-? was transfected into HCC9204, the expression of NF-?B was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-?B is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.

Due to the good tumor-targeting and excellent biocompatibility, the drug-loading nanoparticles (NPs) has been widely applied in the diagnosis and treatment of cancer. However, after the NPs are recognized and internalized by cancer cells, the effects of NPs on cell migration behavior were unclear. In the present study, the self-assembly techniques (SAMs) was used to modify gold (Au) nanoparticles (Au NPs) with different chemical functional groups (CH3, OH, COOH and NH2) as model NPs. The dispersion of these groups in solution and the distribution in cells were studied by transmission electron microscope (TEM), respectively, and the proliferation was examined by MTT assay in vitro. The wound-healing and the Transwell assay were used to examine the effect of internalized Au-NPs on HepG2 cells migration. The results showed that different Au-NPs mainly distributed at the edge of the vesicle membrane and the gap between cells. The Au-NPs resulted in decreased cell viability in a concentration-depended manner. In addition, the results of wound-healing and Transwells assay indicated that the internalization of the NH2-NPs and OH-NPs would inhibit cell migration compared with those in the control group.
Carcinoma, Hepatocellular ; metabolism ; Cell Movement ; Cell Proliferation ; Cell Survival ; Gold ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Metal Nanoparticles ; chemistry

This paper is aimed to investigate the effect of fluid shear stress on the tight junction of laryngeal squamous carcinoma (Hep2) cells and to explore the potential molecular mechanism. Hep2 cells were selected and subjected to the fluid shear stress of 1.4 dyn/cm2 for different time, respectively. The morphological changes of Hep2 cells under shear stress were observed using inverted microscope. The cell-cell junctions were examined by transmission electron microscope (TEM). The expressions of tight junction proteins (including Occludin, Claudin-5 and ZO-1) and the distribution of Claudin-5 were examined by Western blot assay and laser scanning confocal microscope, respectively. The results indicated that Hep2 cells turned to spindle-like shapes after exposed to shear stress, and showed the trend of the recovering to original shapes when the shear stress was cancelled. The cell-cell junctions were tight under the shear flow condition, and the permeability was reduced under the condition of 1.4 dyn/cm shear flow. The expressions of tight junction proteins were enhanced with increased duration of shear flow, but reduced after removing shear flow. The result of Claudin-5 expression by immufluorescence assay was consistent with that by Western blot. The Claudin-5 mainly distributed in the cytoplasm under static condition, while it located at the intercellular after shear flow stimulation, and it appeared intercellular and cytoplasm after stopping shear flow stimulation. Therefore, it can be concluded that shear stress changes the morphology of laryngeal squamous carcinoma Hep2 cells, and upregulates the tight junction.
Blotting, Western ; Carcinoma, Squamous Cell ; pathology ; Claudin-5 ; metabolism ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; pathology ; Occludin ; metabolism ; Stress, Mechanical ; Tight Junctions ; Zonula Occludens-1 Protein ; metabolism