The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (...The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (CNE-2Z cells). Both migrated and non-migrated CNE-2Z cells, when swollen by 47% hypotonic solution, exhibited RVD which was inhibited by extracellular application of chloride channel blockers adenosine 5’-triphosphate (ATP), 5-nitro-2-(3- phenylpropylamino) benzoic acid (NPPB) and tamoxifen. However, RVD rate in migrated CNE-2Z cells was bigger than that of non-migrated cells and the sensitivity of migrated cells to NPPB and tamoxifen was higher than that of non- migrated cells. ATP, NPPB and tamoxifen also inhibited migration of CNE-2Z cells. The inhibition of migration was positively correlated to the blockage of RVD, with a correlation coefficient (r) = 0.99, suggesting a functional relation- ship between RVD and cell migration. We conclude that RVD is involved in cell migration and RVD may play an important role in migratory process in CNE-2Z cells.展开更多
Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human...Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human papillomavirus in HeLa cells is inhibited in the presence of anti-tumor reagent tripchlorolide (TC), which results in the up-regulation of p53 in HeLa cells. Interestingly, under the same TC-treatment, the cells at the early S-phase are more susceptible to apoptosis than those at the middle S-phase although p53 protein is stabilized to the same level in both situations. Significant difference is exhibited between the two specified expression profiles. Further analysis demonstrates that anti-apoptotic gene survivin is up-regulated by p53 in the TC-treated middle-S cells, whereas it is down-regulated by p53 in the TC-treated early-S cells. Taken together, the present study indicates that the differential p53-regulated expres- sion of survivin at different stages of the cell cycle results in different cellular outputs under the same apoptosis-inducer.展开更多
AIM: To evaluate the protective effect of 2′-p-hydroxy benzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl4)-induced toxicity in HUH-7 cel Is.METHODS: CCI4 is a well characterized hep...AIM: To evaluate the protective effect of 2′-p-hydroxy benzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl4)-induced toxicity in HUH-7 cel Is.METHODS: CCI4 is a well characterized hepatotoxin, and inducer of cytochrome P450 2E1 (CYP2E1)-mediated oxidative stress. In addition, lipid peroxidation and accumulation of intracellular calcium are important steps in the pathway involved in CCl4 toxicity. Liver cells (HUH-7) were treated with CCI4, and the mechanism of the cytoprotective effect of NG was assessed. Silymarin, a known hepatoprotective drug, was used as control. RESULTS: NG protected HUH-7 cells against CCl4 toxicity and loss of viability without modulating CYP2E1 activity. Prevention of CCl4 toxicity was associated with a reduction in oxidative damage as reflected by decreased generation of reactive oxygen species (ROS), a decrease in lipid peroxidation and accumulation of intracellular Ca^2+ levels and maintenance of intracellular glutathione homeostasis. Decreased mitochondrial membrane potential (MMP), induction of caspases mediated DNA fragmentation and cell cycle arrest, as a result of CCl4 treatment, were also blocked by NG. The protection afforded by NG seemed to be mediated by activation of cyclic adenosine monophosphate (cAMP) synthesis and inhibition of phospholipases (cPLA2). CONCLUSION: NG exerts a protective effect on CYP2E1-dependent CCl4 toxicity via inhibition of lipid peroxidation, followed by an improved intracellular calcium homeostasis and inhibition of Ca^2+-dependent proteases.展开更多
Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatech...Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatechingallate[EGCG], baikal skullcap root and szechwan lovge rhizome) on HaCaT cells damaged by middle wave ultraviolet(UVB) irradiation. Methods: Subconfluent HaCaT cells were sham or UVB irradiated and treated with above TCM agents. The damage degree of HaCaT cells was observed by a light microscop. Cell growth was recorded by cell count and cellular activity was detected by MTT method. The secretion amount of IL-6 and TNF-α was measured by ELISA. Results: The irradiation damage of HaCaT cells was depended on the irradiated dosages and cellular activity was reduced by 36%-80%, with a maximum decrease over 90% after 72 h. The intervention of the above drugs may increase the cellular activity by 10%-72%. The photo-protective efficiency was more apparent in EGCG (from 1.19±0.07 to 1.28±0.06, P<0.01) than that in hydroxychloroqine (from 0.43±0.04 to 0.96±0.04, P<0.05). The other two tested drugs also showed photo-protective effect(from 0.44±0.07 to 1.21±0.02, P<0.05). As to cytokine secretion, EGCG could decline the secretion amount of IL-6 and TNF-α apparently. Hydroxychloroqine and baikal skullcap root baikal skullcap root could only reduce the secretion of IL-6. The secretion of IL-6 and TNF-α could not be inhibited by szechwan lovge rhizome. Conclusion: The injury effect of UVB irradiation on cultured keratinocytes is dose-dependent and the tested drugs have photo-protective potency. Inhibition of cytokine secretion may be one of the mechanisms related to reducing the damage effect of UVB irradiation.展开更多
AIM: To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N...AIM: To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N) and CCl4-induced hepatic fibrosis model group (group C). After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index (HAI). Histological activity index (HAI), change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS: CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N (P〈0.01). The positive signals decreased significantly in groups T and R (P〈0.01), but positive score was significantly lower in group T than in group R (P〈 0.01). CONCLUS10N: Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ.展开更多
In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To impr...In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture.展开更多
Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) ...Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50 d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigment cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time.展开更多
A continuous marine fish cell line RSBF (i.e. Red Sea Bream Fin) was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine (PEI) and nickel chloride (NiCl 2) in this study on the deleterious effects...A continuous marine fish cell line RSBF (i.e. Red Sea Bream Fin) was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine (PEI) and nickel chloride (NiCl 2) in this study on the deleterious effects of aquatic genotoxins on fish. At the 0.01 to 1 μg/ml concentration tested, PEI had acute toxicity to the treated RSBF cells (IC 50 =1.12, 0.92, 0.88 and 0.64 μg/ml PEI for time 0 h, 24 h, 48 h and 72 h after treatment, respectively) and markedly inhibited their proliferation in a dose dependent manner. At the 0.001 to 5 μmol/L concentration tested, NiCl 2 posed no acute toxicity but significantly stimulated their growth (107%-214% of control). Random amplified polymorphic DNA (RAPD) technique was used to detect the genotoxic effects of PEI and NiCl 2 by comparing the RAPD banding patterns of the control and treated cells. RAPD analysis indicated that at the concentrations tested, PEI was more genotoxic than NiCl 2 to RSBF cells; that there was a slight dose dependent response in the genotoxic effect of PEI but not NiCl 2; and that RAPD technique might provide a sensitive, non specific genotoxic endpoint. And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy.展开更多
AIM: To study the inhibitory effect of mononuclear bone marrow cell (BNC) transplantation on carbon tetrachloride (CCl4) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl4 a...AIM: To study the inhibitory effect of mononuclear bone marrow cell (BNC) transplantation on carbon tetrachloride (CCl4) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl4 and alcohol administration. After 8 wk, twenty rats were randomly allocated into treatment group (n = 10) and control group (n = 10). BMC were infused into the rats in treatment group via the portal vein, while heparinized saline was infused in control group. CCl4 was hypodermically injected into the rats twice a week for 4 wk. At the end of wk 12, all rats were humanely sacrificed. Uver samples were taken and stained with HE or Masson trichrome. The general conditions, liver fibrosis (hydroxyproline and collagen fibre) and liver pathological grades in rats were evaluated. RESULTS: The general conditions of the rats in treatment group improved markedly, but not in control group. Hydroxyproline was 504.6± 128.8 μg/g in treatment group, and 596.0 ± 341.8 μg/g in control group. The percentage of collagen fibre was 3.75% ± 0.98% in treatment group and 5.02% ± 0.44% in control group. There was a significant difference between the two groups (P 〈 0.05). Liver pathological grade decreased from grade N to grade 11 partially in treatment group (P 〈 0.05) with no obvious improvement in control group (P 〉 0.05). There was a significant difference between treatment group and control group (P 〈 0.05).CONCLUSION: Transplantation of BMC can improve liver fibrosis due to chronic liver injury in rats.展开更多
The ultrastructures of skin chloride celis in cultured Japanese flounder and turbot larvae in metamorphosis, which grow in the same feeding conditions, are examined with a transmission electron microscope. These devel...The ultrastructures of skin chloride celis in cultured Japanese flounder and turbot larvae in metamorphosis, which grow in the same feeding conditions, are examined with a transmission electron microscope. These developed skin chloride celis were shaped like flattened ellipsoids and similar in morphology and ultrastructure to typical chloride celis of euryhaline fish gill. They lo-cate in the epidermis and contract with the extra and interior environment through the apical pit and narrow channels. The cytoplasm of cell is full of numerous mitochondria and a ramifying net-work of tubules. The degeneration of skin chloride celis is observed with development of Japanese flounder larvae. Skin chloride celis of turbot are less developmental than those of Japanese flounder in the same developmental stage.展开更多
Salinity is a serious threat to agricultural production. Potato (Solanum tuberosum) is an important food crop characterised for having low to moderate salinity tolerance. Tissue cultures may be relevant to improve s...Salinity is a serious threat to agricultural production. Potato (Solanum tuberosum) is an important food crop characterised for having low to moderate salinity tolerance. Tissue cultures may be relevant to improve salt tolerance in potato through selection of salt-tolerant cell lines and subsequent regeneration of plants. In this work, the authors used the random amplified polymorphic DNA (RAPD) markers to investigate the occurrence of genetic polymorphism in a potato calli line tolerant to 150 mM NaCI. Out of 40 primers screened, eight generated polymorphic patterns that distinguished salt-tolerant line from the control. Although the macroscopic appearance was similar in both lines, ultrastructural study revealed alterations in salt-grown cells. These showed that plastids less differentiated with a lower number of grana had more and larger starch grains than control cells. In conclusion, RAPD analysis revealed that NaCl-adapted line is a somaclonal variant and the ultrastructural study showed changes essentially at the plastids.展开更多
Objective To acquire oval cells (progenitor stem cells ) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley ( SD ) rats were divided into 5 groups randomly: control, 2...Objective To acquire oval cells (progenitor stem cells ) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley ( SD ) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene ( 2-AAF ), 2-AAF + partial hepatectomy ( PH ), 2-AAF + carbon tetrachloride ( CCl4 ), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1. 1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thyl. 1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups : 0. 5 % in 2-AAF, 0. 3% in 2-AAF + PH, 0. 2% in 2-AAF + CCl4, 0. 1% in diabetic, and 0. 0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.展开更多
The occurrence of massive CD4+ T cell depletion is one of the most prominent characteristics of human immunodeficiency virus type 1 (HIV-1) infection during acute phase, resulting in unrestorable destruction to the im...The occurrence of massive CD4+ T cell depletion is one of the most prominent characteristics of human immunodeficiency virus type 1 (HIV-1) infection during acute phase, resulting in unrestorable destruction to the immune system. The infected host undergoes an asymptomatic period lasting several years with low viral load and ostensibly healthy status, which is presumably due to virus-specific adaptive immune responses. In the absence of therapy, an overwhelming majority of cases develop to AIDS within 8-10 years of latent infection. In this review, we discuss the roles in AIDS pathogenesis played by massive CD4+ T lymphocytes depletion in gut-associated lymphoid tissue (GALT) during acute infection and abnormal immune activation emerging in the later part of chronic phase.展开更多
In rice, 2,4-Dichlorphenoxyacetic acid (MS2) has been considered best for callus induction and combination ofNAA, IAA, ABA (MS3) for somatic embryogenesis. Efficient plant regeneration was observed when MS basal s...In rice, 2,4-Dichlorphenoxyacetic acid (MS2) has been considered best for callus induction and combination ofNAA, IAA, ABA (MS3) for somatic embryogenesis. Efficient plant regeneration was observed when MS basal salts supplied with 3% sorbitol and 3% maltose as carbon sources without hormones (MSac). During this experiment, effect of sodium chloride (NaCl) and abscisic acid (ABA) was assessed on callus growth, plant regeneration and root induction efficiency of rice (Oryza sativa L.) varieties IR-6 and Basmati-370. Callus proliferation rate was highly decreased in both varieties on MS2b (100 mol.m3 NaCl + 0.5 mg'L1 ABA) than MS2a (100 mol'm3 NaCl) cultures significantly. Proline, glycine-betaine and reducing sugars were increased significantly in MS2a and MS2b callusing cultures. However, total proteins were decreased in MS2a, while slightly increased in MS2b. Maximum plant regeneration (9.42 ±0.54 and 10.67 ±0.50 plantlets.callus1) from somatic embryos was observed on MS4c in IR-6 and Basmati-370, while 1.56 ± 0.06 (IR-6) and 0.95 ±0.05 (Basmati-370) plantlets callus1 were observed on MSab (100 mol·m3 NaCl). No plant regeneration was observed on MS4b (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) medium in both varieties. Inhibition of root induction efficiency was high in MSsb (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) than MS5a (100 mol·m3 NaCl) in the stressed cultures (P 〈 0.05). In this experiment, it was concluded that ABA involved in somatic embryogenesis and elevation of NaCl stress, while it causes inhibition of cell's growth as well as its regeneration into plantlets from somatic embryos.展开更多
Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 c...Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 ceils impaired by COCl2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 ceils exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. Results COCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. COCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 μmol/L. Conclusion These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities.展开更多
In the present study, we investigated the activation of protein kinase C (PKC) family in mouse embryonic fibroblast NIH3T3 cells using gadolinium chloride as a representative lanthanide ion. With live cell imaging s...In the present study, we investigated the activation of protein kinase C (PKC) family in mouse embryonic fibroblast NIH3T3 cells using gadolinium chloride as a representative lanthanide ion. With live cell imaging system and confocal laser scanning microscopy, we found that the treatment of 50 μM GdCI3 promoted cell survival under the condition of serum-starvation. Moreover, better cell attachment and cytoskeleton reorganization were also observed. Additionally, GdC13 treatment resulted in the phosphorylation of PKC family at different time points. Furthermore, bisindolylmaleimide (a PKCpan inhibitor) could efficiently reduce the level of phosphorylated PKCpan (βIISer660), alleviating ERK activation induced by GdC13. This finding indicated that the PKC activation was involved in GdC13-induced MAPK/ERK signaling and thus might contribute to GdClβ-indueed cell cycle progression and cell survival.展开更多
The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands.Using a hydroponic culture system,the secretion and accumulation of Na+ and K+ in Tamarix ramosissima and T...The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands.Using a hydroponic culture system,the secretion and accumulation of Na+ and K+ in Tamarix ramosissima and T.laxa under different salt stresses (NaCl,KCl and NaCl+KCl) were studied.Additionally,the effects of salt gland inhibitors (orthovanadate,Ba2+,ouabain,tetraethylammonium (TEA) and verapamil) on Na+ and K+ secretion and accumulation were examined.Treatment with NaCl (at 0-200 mmol L 1 levels) significantly increased Na+ secretion,whereas KCl treatment (at 0-200 mmol L 1 levels) significantly increased K+ secretion.The ratio of secretion to accumulation of Na+ was higher than that of K+.The changes in Na+ and K+ secretion differed after adding different ions into the single-salt solutions.Addition of NaCl to the KCl solution (at 100 mmol L 1 level,respectively) led to a significant decrease in K+ secretion rate,whereas addition of KCl to the NaCl solution (at 100 mmol L 1 level,respectively) had little impact on the Na+ secretion rate.These results indicated that Na+ secretion in Tamarix was highly selective.In addition,Na+ secretion was significantly inhibited by orthovanadate,ouabain,TEA and verapamil,and K+ secretion was significantly inhibited by ouabain,TEA and verapamil.The different impacts of orthovanadate on Na+ and K+ secretion might be the primary cause for the different Na+ and K+ secretion abilities of multicellular salt glands in Tamarix.展开更多
基金This work was supported by grants from the Wellcome Trust UK(056909/299/Z)Ministry of Education of China(GJ9901)+1 种基金the Health Department of Guangdong Province(A2001474)Guangdong Medical College Fund for Young Scholars(XQ0306).
文摘The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (CNE-2Z cells). Both migrated and non-migrated CNE-2Z cells, when swollen by 47% hypotonic solution, exhibited RVD which was inhibited by extracellular application of chloride channel blockers adenosine 5’-triphosphate (ATP), 5-nitro-2-(3- phenylpropylamino) benzoic acid (NPPB) and tamoxifen. However, RVD rate in migrated CNE-2Z cells was bigger than that of non-migrated cells and the sensitivity of migrated cells to NPPB and tamoxifen was higher than that of non- migrated cells. ATP, NPPB and tamoxifen also inhibited migration of CNE-2Z cells. The inhibition of migration was positively correlated to the blockage of RVD, with a correlation coefficient (r) = 0.99, suggesting a functional relation- ship between RVD and cell migration. We conclude that RVD is involved in cell migration and RVD may play an important role in migratory process in CNE-2Z cells.
基金This work was supported by a grant of National Natu-ral Science Foundation of China(No.30230110)a grant of Science and Technology Committee of Shanghai Mu-nicipality(No.04DZ14901)a grant of Chinese Acad-emy of Sciences#KSCX2-SW-203 to Jia Rui WU.
文摘Recent studies indicate that cell-cycle checkpoints are tightly correlated with the regulation of apoptosis, in which p53 plays an important role. Our present works show that the expression of E6/E7 oncogenes of human papillomavirus in HeLa cells is inhibited in the presence of anti-tumor reagent tripchlorolide (TC), which results in the up-regulation of p53 in HeLa cells. Interestingly, under the same TC-treatment, the cells at the early S-phase are more susceptible to apoptosis than those at the middle S-phase although p53 protein is stabilized to the same level in both situations. Significant difference is exhibited between the two specified expression profiles. Further analysis demonstrates that anti-apoptotic gene survivin is up-regulated by p53 in the TC-treated middle-S cells, whereas it is down-regulated by p53 in the TC-treated early-S cells. Taken together, the present study indicates that the differential p53-regulated expres- sion of survivin at different stages of the cell cycle results in different cellular outputs under the same apoptosis-inducer.
基金Indian Institute of Integrative Medicine, Council of Scientific and Industrial Research
文摘AIM: To evaluate the protective effect of 2′-p-hydroxy benzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl4)-induced toxicity in HUH-7 cel Is.METHODS: CCI4 is a well characterized hepatotoxin, and inducer of cytochrome P450 2E1 (CYP2E1)-mediated oxidative stress. In addition, lipid peroxidation and accumulation of intracellular calcium are important steps in the pathway involved in CCl4 toxicity. Liver cells (HUH-7) were treated with CCI4, and the mechanism of the cytoprotective effect of NG was assessed. Silymarin, a known hepatoprotective drug, was used as control. RESULTS: NG protected HUH-7 cells against CCl4 toxicity and loss of viability without modulating CYP2E1 activity. Prevention of CCl4 toxicity was associated with a reduction in oxidative damage as reflected by decreased generation of reactive oxygen species (ROS), a decrease in lipid peroxidation and accumulation of intracellular Ca^2+ levels and maintenance of intracellular glutathione homeostasis. Decreased mitochondrial membrane potential (MMP), induction of caspases mediated DNA fragmentation and cell cycle arrest, as a result of CCl4 treatment, were also blocked by NG. The protection afforded by NG seemed to be mediated by activation of cyclic adenosine monophosphate (cAMP) synthesis and inhibition of phospholipases (cPLA2). CONCLUSION: NG exerts a protective effect on CYP2E1-dependent CCl4 toxicity via inhibition of lipid peroxidation, followed by an improved intracellular calcium homeostasis and inhibition of Ca^2+-dependent proteases.
文摘Objective: To investigate damage effects of ultraviolet irradiation on eternal keratinocyte-HaCaT cells and to evaluate photo-protective efficiency of hydroxychloroqine and Traditional Chinese Medicines(epigallocatechingallate[EGCG], baikal skullcap root and szechwan lovge rhizome) on HaCaT cells damaged by middle wave ultraviolet(UVB) irradiation. Methods: Subconfluent HaCaT cells were sham or UVB irradiated and treated with above TCM agents. The damage degree of HaCaT cells was observed by a light microscop. Cell growth was recorded by cell count and cellular activity was detected by MTT method. The secretion amount of IL-6 and TNF-α was measured by ELISA. Results: The irradiation damage of HaCaT cells was depended on the irradiated dosages and cellular activity was reduced by 36%-80%, with a maximum decrease over 90% after 72 h. The intervention of the above drugs may increase the cellular activity by 10%-72%. The photo-protective efficiency was more apparent in EGCG (from 1.19±0.07 to 1.28±0.06, P<0.01) than that in hydroxychloroqine (from 0.43±0.04 to 0.96±0.04, P<0.05). The other two tested drugs also showed photo-protective effect(from 0.44±0.07 to 1.21±0.02, P<0.05). As to cytokine secretion, EGCG could decline the secretion amount of IL-6 and TNF-α apparently. Hydroxychloroqine and baikal skullcap root baikal skullcap root could only reduce the secretion of IL-6. The secretion of IL-6 and TNF-α could not be inhibited by szechwan lovge rhizome. Conclusion: The injury effect of UVB irradiation on cultured keratinocytes is dose-dependent and the tested drugs have photo-protective potency. Inhibition of cytokine secretion may be one of the mechanisms related to reducing the damage effect of UVB irradiation.
基金Supported by Nature Science Foundation of Fujian Province. No.2005D094 and No.C0410025
文摘AIM: To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N) and CCl4-induced hepatic fibrosis model group (group C). After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index (HAI). Histological activity index (HAI), change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS: CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N (P〈0.01). The positive signals decreased significantly in groups T and R (P〈0.01), but positive score was significantly lower in group T than in group R (P〈 0.01). CONCLUS10N: Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ.
基金Supported by the National Basic Research Program of China(2007CB707805) the National Natural Science Foundation of China(20876139)
文摘In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture.
基金funded by the NNSF of China(No.30070593)SNSF(No.Y2002D10).
文摘Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50 d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigment cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time.
文摘A continuous marine fish cell line RSBF (i.e. Red Sea Bream Fin) was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine (PEI) and nickel chloride (NiCl 2) in this study on the deleterious effects of aquatic genotoxins on fish. At the 0.01 to 1 μg/ml concentration tested, PEI had acute toxicity to the treated RSBF cells (IC 50 =1.12, 0.92, 0.88 and 0.64 μg/ml PEI for time 0 h, 24 h, 48 h and 72 h after treatment, respectively) and markedly inhibited their proliferation in a dose dependent manner. At the 0.001 to 5 μmol/L concentration tested, NiCl 2 posed no acute toxicity but significantly stimulated their growth (107%-214% of control). Random amplified polymorphic DNA (RAPD) technique was used to detect the genotoxic effects of PEI and NiCl 2 by comparing the RAPD banding patterns of the control and treated cells. RAPD analysis indicated that at the concentrations tested, PEI was more genotoxic than NiCl 2 to RSBF cells; that there was a slight dose dependent response in the genotoxic effect of PEI but not NiCl 2; and that RAPD technique might provide a sensitive, non specific genotoxic endpoint. And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy.
基金the Key Scientific and Technological Projects of Guangdong Province, No. 2005A30201007Guangdong Provincial Natural Science Fundation, No. 06021343
文摘AIM: To study the inhibitory effect of mononuclear bone marrow cell (BNC) transplantation on carbon tetrachloride (CCl4) -induced liver fibrosis in rats. METHODS: Rat liver fibrosis models were induced by CCl4 and alcohol administration. After 8 wk, twenty rats were randomly allocated into treatment group (n = 10) and control group (n = 10). BMC were infused into the rats in treatment group via the portal vein, while heparinized saline was infused in control group. CCl4 was hypodermically injected into the rats twice a week for 4 wk. At the end of wk 12, all rats were humanely sacrificed. Uver samples were taken and stained with HE or Masson trichrome. The general conditions, liver fibrosis (hydroxyproline and collagen fibre) and liver pathological grades in rats were evaluated. RESULTS: The general conditions of the rats in treatment group improved markedly, but not in control group. Hydroxyproline was 504.6± 128.8 μg/g in treatment group, and 596.0 ± 341.8 μg/g in control group. The percentage of collagen fibre was 3.75% ± 0.98% in treatment group and 5.02% ± 0.44% in control group. There was a significant difference between the two groups (P 〈 0.05). Liver pathological grade decreased from grade N to grade 11 partially in treatment group (P 〈 0.05) with no obvious improvement in control group (P 〉 0.05). There was a significant difference between treatment group and control group (P 〈 0.05).CONCLUSION: Transplantation of BMC can improve liver fibrosis due to chronic liver injury in rats.
文摘The ultrastructures of skin chloride celis in cultured Japanese flounder and turbot larvae in metamorphosis, which grow in the same feeding conditions, are examined with a transmission electron microscope. These developed skin chloride celis were shaped like flattened ellipsoids and similar in morphology and ultrastructure to typical chloride celis of euryhaline fish gill. They lo-cate in the epidermis and contract with the extra and interior environment through the apical pit and narrow channels. The cytoplasm of cell is full of numerous mitochondria and a ramifying net-work of tubules. The degeneration of skin chloride celis is observed with development of Japanese flounder larvae. Skin chloride celis of turbot are less developmental than those of Japanese flounder in the same developmental stage.
文摘Salinity is a serious threat to agricultural production. Potato (Solanum tuberosum) is an important food crop characterised for having low to moderate salinity tolerance. Tissue cultures may be relevant to improve salt tolerance in potato through selection of salt-tolerant cell lines and subsequent regeneration of plants. In this work, the authors used the random amplified polymorphic DNA (RAPD) markers to investigate the occurrence of genetic polymorphism in a potato calli line tolerant to 150 mM NaCI. Out of 40 primers screened, eight generated polymorphic patterns that distinguished salt-tolerant line from the control. Although the macroscopic appearance was similar in both lines, ultrastructural study revealed alterations in salt-grown cells. These showed that plastids less differentiated with a lower number of grana had more and larger starch grains than control cells. In conclusion, RAPD analysis revealed that NaCl-adapted line is a somaclonal variant and the ultrastructural study showed changes essentially at the plastids.
基金Supported by Shanghai Municipal Education Commission Key Project (07ZZ35)
文摘Objective To acquire oval cells (progenitor stem cells ) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley ( SD ) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene ( 2-AAF ), 2-AAF + partial hepatectomy ( PH ), 2-AAF + carbon tetrachloride ( CCl4 ), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1. 1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thyl. 1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups : 0. 5 % in 2-AAF, 0. 3% in 2-AAF + PH, 0. 2% in 2-AAF + CCl4, 0. 1% in diabetic, and 0. 0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.
文摘The occurrence of massive CD4+ T cell depletion is one of the most prominent characteristics of human immunodeficiency virus type 1 (HIV-1) infection during acute phase, resulting in unrestorable destruction to the immune system. The infected host undergoes an asymptomatic period lasting several years with low viral load and ostensibly healthy status, which is presumably due to virus-specific adaptive immune responses. In the absence of therapy, an overwhelming majority of cases develop to AIDS within 8-10 years of latent infection. In this review, we discuss the roles in AIDS pathogenesis played by massive CD4+ T lymphocytes depletion in gut-associated lymphoid tissue (GALT) during acute infection and abnormal immune activation emerging in the later part of chronic phase.
文摘In rice, 2,4-Dichlorphenoxyacetic acid (MS2) has been considered best for callus induction and combination ofNAA, IAA, ABA (MS3) for somatic embryogenesis. Efficient plant regeneration was observed when MS basal salts supplied with 3% sorbitol and 3% maltose as carbon sources without hormones (MSac). During this experiment, effect of sodium chloride (NaCl) and abscisic acid (ABA) was assessed on callus growth, plant regeneration and root induction efficiency of rice (Oryza sativa L.) varieties IR-6 and Basmati-370. Callus proliferation rate was highly decreased in both varieties on MS2b (100 mol.m3 NaCl + 0.5 mg'L1 ABA) than MS2a (100 mol'm3 NaCl) cultures significantly. Proline, glycine-betaine and reducing sugars were increased significantly in MS2a and MS2b callusing cultures. However, total proteins were decreased in MS2a, while slightly increased in MS2b. Maximum plant regeneration (9.42 ±0.54 and 10.67 ±0.50 plantlets.callus1) from somatic embryos was observed on MS4c in IR-6 and Basmati-370, while 1.56 ± 0.06 (IR-6) and 0.95 ±0.05 (Basmati-370) plantlets callus1 were observed on MSab (100 mol·m3 NaCl). No plant regeneration was observed on MS4b (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) medium in both varieties. Inhibition of root induction efficiency was high in MSsb (100 mol·m3 NaCl + 0.5 mg·L-1 ABA) than MS5a (100 mol·m3 NaCl) in the stressed cultures (P 〈 0.05). In this experiment, it was concluded that ABA involved in somatic embryogenesis and elevation of NaCl stress, while it causes inhibition of cell's growth as well as its regeneration into plantlets from somatic embryos.
文摘Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 ceils impaired by COCl2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 ceils exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. Results COCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. COCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 μmol/L. Conclusion These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities.
基金National Natural Science Foundation of China (Grant No.21277006 and 20637010)
文摘In the present study, we investigated the activation of protein kinase C (PKC) family in mouse embryonic fibroblast NIH3T3 cells using gadolinium chloride as a representative lanthanide ion. With live cell imaging system and confocal laser scanning microscopy, we found that the treatment of 50 μM GdCI3 promoted cell survival under the condition of serum-starvation. Moreover, better cell attachment and cytoskeleton reorganization were also observed. Additionally, GdC13 treatment resulted in the phosphorylation of PKC family at different time points. Furthermore, bisindolylmaleimide (a PKCpan inhibitor) could efficiently reduce the level of phosphorylated PKCpan (βIISer660), alleviating ERK activation induced by GdC13. This finding indicated that the PKC activation was involved in GdC13-induced MAPK/ERK signaling and thus might contribute to GdClβ-indueed cell cycle progression and cell survival.
基金supported by the Special Fund for Public Welfare Industrial(Agriculture) Research of China(Grant No. 200903001)Chinese Academy of Sciences Innovation Base Project for Agriculture(Grant No. KSCX1-YW-09-10),the Chinese Academy of Sciences Agricultural Projects (Grant No. KSCX2-YW-N-41)
文摘The present study aimed to determine the mechanism of cation-selective secretion by multicellular salt glands.Using a hydroponic culture system,the secretion and accumulation of Na+ and K+ in Tamarix ramosissima and T.laxa under different salt stresses (NaCl,KCl and NaCl+KCl) were studied.Additionally,the effects of salt gland inhibitors (orthovanadate,Ba2+,ouabain,tetraethylammonium (TEA) and verapamil) on Na+ and K+ secretion and accumulation were examined.Treatment with NaCl (at 0-200 mmol L 1 levels) significantly increased Na+ secretion,whereas KCl treatment (at 0-200 mmol L 1 levels) significantly increased K+ secretion.The ratio of secretion to accumulation of Na+ was higher than that of K+.The changes in Na+ and K+ secretion differed after adding different ions into the single-salt solutions.Addition of NaCl to the KCl solution (at 100 mmol L 1 level,respectively) led to a significant decrease in K+ secretion rate,whereas addition of KCl to the NaCl solution (at 100 mmol L 1 level,respectively) had little impact on the Na+ secretion rate.These results indicated that Na+ secretion in Tamarix was highly selective.In addition,Na+ secretion was significantly inhibited by orthovanadate,ouabain,TEA and verapamil,and K+ secretion was significantly inhibited by ouabain,TEA and verapamil.The different impacts of orthovanadate on Na+ and K+ secretion might be the primary cause for the different Na+ and K+ secretion abilities of multicellular salt glands in Tamarix.