1.Rat prostate glandular epithelial cells cultured in vitro and their barrier function.
Dong CUI ; Yong-gang SHANG ; Guang-wei HAN ; Cheng-cheng LIU ; Shan-hong YI
National Journal of Andrology 2016;22(2):133-137
OBJECTIVETo culture rat prostate glandular epithelial cells and study their barrier functions in vitro.
METHODSRat prostate glandular epithelial cells were cultured in vitro. The expression of the tight junction protein claudin-1 was determined by immunohistochemistry, the structure and composition of the epithelial cells observed under the inverted microscope and transmission electron microscope. The transepithelial electrical resistances (TEERs) were monitored with the Millicell system. The permeability of the prostate glandular epithelial cells was assessed by the phenol red leakage test.
RESULTSCompact monolayer cell structures were formed in the prostate glandular epithelial cells cultured in vitro. Immunohistochemistry showed the expression of the tight junction protein claudin-1 and transmission electron microscopy confirmed the formation of tight junctions between the adjacent glandular epithelial cells. The TEERs in the cultured prostate glandular epithelial cells reached the peak of about (201.3 ± 3.5) Ω/cm2 on the 8th day. The phenol red leakage test manifested a decreased permeability of the cell layers with the increase of TEERs.
CONCLUSIONThe structure and function of rat prostate glandular epithelial cells are similar to those of brain capillary endothelial cells, retinal capillary endothelial cells, and intestinal epithelial cells. In vitro cultured prostate glandular epithelial cells have the barrier function and can be used as a model for the study of blood prostate barrier in vitro.
Animals ; Cell Membrane Permeability ; Cells, Cultured ; Claudin-1 ; metabolism ; Electric Impedance ; Epithelial Cells ; pathology ; physiology ; ultrastructure ; In Vitro Techniques ; Male ; Microscopy, Electron, Transmission ; Phenolsulfonphthalein ; pharmacokinetics ; Prostate ; metabolism ; pathology ; Rats ; Tight Junctions
2.Over-expression of hypoxia-inducible factor-1alpha induces epithelial-mesenchymal transition in LNCaP cells in vitro and in vivo.
Yi-li HAN ; Yong LUO ; Da-lin HE ; Yong-yi CHENG ; Yong-gang XU
National Journal of Andrology 2012;18(1):23-28
OBJECTIVETo investigate the effects of the over-expression of hypoxia-inducible factor-1alpha (HIF-1alpha) on the epithelial-mesenchymal transition (EMT) and invasive potency of LNCaP cells in vitro and in vivo.
METHODSWe cultured LNCaP cells stably expressing HIF-1alpha (LNCaP/HIF-1alpha) and LNCaP cells, identified the over-expression of HIF-1alpha, determined the proliferation of the two cell lines by MTT assay and the level of PSA in the supernatant of culture medium, and detected the anchorage independent growth by soft-agar colony formation assay. A subcutaneous tumor model was established in nude mice by injecting LNCaP/HIF-1alpha and LNCaP cells followed by observation of the tumor growth. Tumor specimens were obtained for immunohistochemistry.
RESULTSThe over-expression of HIF-1alpha was confirmed in the LNCaP/HIF-1alpha cells by immunofluorescence staining and Western blotting. The level of PSA was obviously decreased in LNCaP/HIF-1alpha as compared with that in LNCaP cells. MTT assay identified the increased proliferation of LNCaP/HIF-1alpha cells. The cell colony forming ability of the LNCaP cells was significantly lower than that of the LNCaP/HIF-1alpha cells. The rate of tumorigenesis was increased and its time shortened in the LNCaP/HIF-1alpha group. Immunohistochemistry revealed an up-regulated expression of vimentin and a down-regulated expression of E-cadherin in the tumor specimens.
CONCLUSIONThe overexpression of HIF-1alpha can up-regulate the expression of vimentin and down-regulate the expression of E-cadherin, which may enhance the invasive potency of LNCaP cells by inducing EMT.
Animals ; Cell Hypoxia ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; metabolism ; pathology
3.Analysis of pathological changes of bone hydatid cyst of meriones meridianus after radiotherapy
Liang, CHEN ; Cheng-peng, YI ; Qi-xin, XIE ; Yong-ming, CHEN ; Yao, ZHANG ; Zeng-ru, XIE
Chinese Journal of Endemiology 2013;32(4):389-394
Objective To observe the pathological changes of bone hydatid cyst of meriones meridianus after radiation therapy,and to investigate the clinical effect of radiotherapy on bone hydatid disease.Methods Ascus was dissected sterilely from sheep liver that naturally infected with Echinococcus granulomas,sheared and sac skin removed.Then it was washed and precipitated with 0.9% sterile saline for 3 times,and scolex was HE stained and counted,from which a 20 ml suspension was made containing 12 × 106/L of scolex.Health meriones meridianus (referred to as gerbil) 140,male and female were in each half,aged 2 to 3 months,body weight(38 ± 6)g,were involved in the study.Gerbil was injected a 0.2 ml suspension containing Echinococcus granulomas scolex into hind tibial periosteum,and X-ray was taken 12 months after the injection.According to the bone destruction in the vaccination site,gerbil hindleg tibia with clear jagged bone destruction was treated as inclusion criteria,and 72 animal were selected as gerbil bone hydatid disease animal models,male and female were in each half.A tatal of 72 gerbils were randomly divided into 4 groups:control group,40 beequerel(Gy) group,50 Gy group and 60 Gy group,18 rats in each group,male and female in each half.The model animals were treated with radiotherapy for 5 times,with 2 d interval,and radiation dose was 300 cGy/min.Each group of gerbils was sacrificed after radiotherapy,bone Echinococcus granulomas cysts was taken out sterilely,and observed by light and electron microscope.Intracapsular cyst fluid was extracted,washed and precipitated with 0.9% sterile saline repeatedly,and and the pellet was HE stained for observation of scolex morphology and activity by light microscope.Results The morphology and activity ofEchinococcus granulomas in cystic fluid in control group were normal; the morphology and activity of Echinococcus granulomas were still normal in the 40 Gy group,and Echinococcus granulomas was not stained red; but those were abnormal,deformation and atrophy and stained red in the 50 Gy group; and were stained red,deformed,fractured and were wrapped by unknown in the 60 Gy group.By light microscope,the germinal layer,cuticle layer,brood capsule and histological structure of protoscolex were basically normal in irradiated region in the control group.The pathological changes of hydatid cyst in the 40 Gy group were mainly degeneration,structure of hydatid cyst was abnormal,stratum corneum was extensive edema,germinal layer became thinner and the fertile cyst was rare.The main pathological change of hydatid cyst in the 50 Gy group was that corneous layer was widely fractured,and the germinal layer was edema,buckling folds,cells decreased,rare seen brood capsule and scolex; the main pathological changes of hydatid cyst were mainly necrosis in the 60 Gy group,cuticle was extensive fault,stratum corneum and germinal layer was separated,germinal layer was atrophy and disorder,no brood capsule and scolex.By electron microscope,cuticle structure of Echinococcus granulomas cyst was clear,microvillus arranged neatly,morphology and structure of the cell and organelle in cytoplasm were normal in the control group.There were many inflammatory cells infiltrating germinal layer of Echinococcus granulomas cyst,microfilament and contents in microfilament were reduced in the 40 Gy group.Microvillus of Echinococcus granulomas disappeared,nuclear membrane was unclear,endoplasmic and mitochon eclasis,lymphocyte nuclear chromatin was clumping and edge set and in circular permutation in the 50 Gy group.Microvillus disappeared,perinuclear membrane indistinct and ruptured,parts of nucleoli were fragmented and marinated,endoplasmic reticulum was extensive expansion,mitochondria was pyknosis and obvious vacuolization,lymphocyte nuclear chromatin clumping and edge set,lysosomes and macrophage emerge in the 60 Gy group.Conclusions Radiotherapy can destroy the morphology and structure of bone hydatid cyst,radioactivity at 50 Gy has a lethal effect on hydatid cyst.Radiation treatment of bone hydatid disease has a good clinical effect.
4.Preparation and in vitro evaluation of doxorubicin-loaded magnetic iron oxide nanoparticles.
Song SHEN ; Lin WU ; Cheng-Run WANG ; Xue-Yong QI ; Yan-Ru GE ; Yi JIN
Acta Pharmaceutica Sinica 2013;48(12):1844-1849
PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.
Antibiotics, Antineoplastic
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administration & dosage
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pharmacology
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Cell Survival
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drug effects
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Doxorubicin
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administration & dosage
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pharmacology
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Drug Carriers
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Ferrosoferric Oxide
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chemistry
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Humans
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Hyperthermia, Induced
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MCF-7 Cells
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Magnetite Nanoparticles
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chemistry
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Particle Size
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Polyethylene Glycols
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chemistry
5.The protection of vitamin E on LTP in hippocampal dentate gyrus of rats under stress.
Yan HONG ; Yi-Yong CHENG ; Qiang MA
Chinese Journal of Applied Physiology 2002;18(2):142-144
AIMTo study the effects of vitamin E on stress-induced impairments in hippocampus of rats.
METHODSTwenty four male Wistar rats were randomly allocated into four groups: control, stress, control+ VE, stress+ VE. The rat stress model was built by restraining for 6 h/d,21 d. The long-term potentiation was induced in rat hippocampal dentate gyrus (DG) by high-frequency test stimulation.
RESULTSCompared with control group, the rats suffered from restraint stress showed that the number of crossing in open-field test and the content of glucocorticoids in plasma was significantly increased, the changes of amplitude of population spike (PS) were significantly lower. After VE supplementation in stress rats, the indices mentioned above were significantly improved.
CONCLUSIONOpportune supplementation of vitamin E may improve the brain function under stress.
Animals ; Dentate Gyrus ; drug effects ; physiology ; Electric Stimulation ; Hippocampus ; drug effects ; physiology ; Long-Term Potentiation ; drug effects ; Male ; Rats ; Rats, Wistar ; Stress, Physiological ; Vitamin E ; pharmacology
6.Expressions of aquaporins decrease in the prostate and seminal vesicles of castrated rats.
Jun JIANG ; Jin-cheng TIAN ; Ji-yi XIA ; Yong-sheng ZHU ; Rui JIANG
National Journal of Andrology 2015;21(4):300-307
OBJECTIVETo investigate the levels of secretions from the prostate and seminal vesicles and their association with the expressions of aquaporins (AQP) in the prostatic tissue and seminal vesicles of castrated rats.
METHODSWe randomly divided 18 eight-week-old male SD rats into a control, a castration, and a testosterone (T) replacement group. Four weeks after surgical castration, we detected the plasma T level and measured the volumes of the secretions and the expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the rats.
RESULTSThe plasma T level was significantly lower in the castrated models ([30. 98 ± 28. 84] ng/dl) than in the rats of the control ([700.78 ± 123.8] ng/dl) and T replacement groups ([688.08 ± 132. 47] ng/dl) (P <0. 05). The castration group, in comparison with the control and T replacement groups, showed remarkably reduced ratios of prostatic secretion volume / prostate weight ([11.1 ± 0.30] vs [2.32 ± 0.61] and [2.13 ± 0.56] %, P <0. 05) and seminal vesicle secretion volume / seminal vesicle weight ( [4. 78 ± 1. 97 ] vs [57. 36 ± 11. 86] and [55. 74 ± 7. 21] %, P < 0. 05). Immunohistochemistry revealed the expressions of AQPs 3 and 7 in the epithelial envelop and cytoplasm and that of AQP 11 the in endothelial envelop and cytoplasm of the prostate and seminal vesicles. Western blot exhibited significantly lower expressions of AQPs 3, 7, and 10 - 12 in the prostate and seminal vesicles of the castrated rats than in the animals of the control and T replacement groups (P <0. 05).
CONCLUSIONSignificant decreases of the secretions from the prostate and seminal vesicles may be related to the reduced expressions of AQPs 3, 7, and 10 - 12 in the prostatic tissue and seminal vesicles in castrated rats.
Animals ; Aquaporins ; metabolism ; Humans ; Male ; Orchiectomy ; Prostate ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminal Vesicles ; metabolism ; Testosterone ; blood
7.Study on BALB/c mice by immunization with genetic engineering vaccines of enterohaemorrhagic Escherichia coil O157:117
Jianping CHENG ; Quanming ZOU ; Xuhu MAO ; Yong YI ; Qingxu WANG ; Ying MA
Chinese Journal of Microbiology and Immunology 2008;28(9):828-832
Objective To investigate immunoprophylactic potential of genetic engineering vaccines of enterohaemorrhagie Escherichia coli O157:H7 in BALB/c mice after immunization with these vaccines. Methods Sixty BALB/c mice (3 weeks old) were randomized averagely into 5 groups. Group 1-3 were im-munized respectively with IntiminC300, Stx2B and HIyAN436, group 4 with a combination of these three vaccines, and group 5 with PBS. Each mouse was immunized with vaccine(100 μg)and Al(OH)3 adjuvant (100 μg) for 3 times. After 7 d of the second and third immunization, serum of each mouse was collected and the different antibodies were detected. After 10 d of the last immunization, all mice were given drinking water containing streptomycin for 3 d before and following oral challenge with O157:H7 (109 CFU), and treated with clinical, microbiological and pathological examination. Results The three vaccines elicited high titer antiserum, and some mice were died after infection with O157. The livability of group 1-4 was re-spectively 73%, 64%, 36% and 91%. And these vaccines depressed fecal and colon shedding with O157. Condusion IntiminC300, Stx2B and HIyAN436 have certain protective efficacy for infection of O157, and combined immunization was more effective than single vaccine.
8.Induction of Endogenous Neural Stem Cells with Hyperbaric Oxygen after Half Cut-off of Spinal Cord in Rats
Hai LIU ; Zhong-cheng WANG ; Yi-hua AN ; Yong CUI ; Qiang JIN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(5):369-371
ObjectiveTo investigate the effect of hyperbaric oxygen (HBO) on proliferation and differentiation of endogenous neural stem cells after acute spinal cord half cut-off in rats. MethodsThe differences of proliferation and differentiation of endogenous neural stem cells between injured group and intervention group were compared. ResultsThere were remarkable differences between injured group and intervention group. ConclusionHBO can promote the proliferation and differentiation of the neural stem cells in rats after spine cord injury.
9.Prevalence and associated factors of school physical violence behaviors among middle school students in Beijing
Yi-Juan QIAO ; Yi XING ; Jia-Li DUAN ; Cheng-Xu BAI ; Yong-Ping PAN ; Yong-Qiang CUI ; Jun-Hua KONG
Chinese Journal of Epidemiology 2010;31(5):510-512
Objective To described the prevalence of school physical violence behaviors and to explore its associated factors among middle school students in Beijing. Methods In 2009, a randomly selected cross- sectional survey was conducted among 5718 students in grades 7 to 12 in Beijing. A self-report anonymous questionnaire involving physical violence at school and sociodemographic variables, such as sex, grades, family economic status and family structure, peer relationships, and communication with their parents etc. were completed by students themselves.Logistic regression was used to estimate the association between physical violence and sociodemographic variables. Results Among the students, 14.3% reported that they had had physical violence behavior in school during the past 12 months. Male students had been more likely to have physical violence behaviors than female students (Male 25.2%, Female 5.1% ). For both male and female students, poor school cohesion were the risk factors of physical violence behaviors (Male OR=1.060, Female OR=1.065). For male students, factors as father' s lower education level (OR=1.653 ), remarried/single-parent families ( OR = 1.834 ), low-grade ( grade 7 OR = 5.291; grade 11 OR =1.526) , poor school performance (OR=1.470) etc were the risk factors of physical violence behaviors; while better-off family economic status (OR=0.546), good peer relationships (OR=0.618) , and easy to communicate with the father (OR=0.756) were the protective factors of physical violence behaviors. For female students, easy to communicate with her mother (OR = 0.358)were the protective factors of physical violence behaviors. Conclusion For male and female students, the prevalence of school physical violence and its related factors were different. Actions on prevention against physical violence behaviors should be fully considered, including factors as gender, personal characteristics, family, school and peers etc.
10.Study on the solubility changes of Rhizoma Coptidis' main chemical constituents influenced by the proportion of Rhizoma Coptidis and Fructus Evodiae.
Ming-xing PENG ; Yong-jiang WU ; Yi-yu CHENG
China Journal of Chinese Materia Medica 2003;28(7):629-632
OBJECTIVETo investigate the relationship between the solubility of Rhizoma Coptidis' main chemical constituents and the proportion of Rhizoma Coptidis and Fructus Evodiae.
METHOD9 samples were selected in which the proportion of Rhizoma Coptidis and Fructus Evodiae were 1:0, 6:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:6 and 0:1, respectively. The samples' water-extracts were analyzed by RP-HPLC. Then solubility changes of eight chemical constituents were calculated, and the regression equations were established.
RESULTAmong the eight chemical constituents, one was independent on the change of proportion of Rhizoma Coptidis and Fructus Evodiae, six having linear relationship. A new constituent was detected, and there was nonlinear relationship between its quantity and the proportion of the couple.
CONCLUSIONThe solubility of Rhizoma Coptidis' main chemical constituents declines almost in the same scale when Rhizoma Coptidis and Fructus Evodiae are used as a couple, and a new constituent has been detected which can't be found in the mono-extraction.
Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; Coptis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; Evodia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Solubility