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OBJECTIVE: To study the chemical constituents from Polygonum cuspidatum. METHODS: The petroleum ether-ethyl acetate solvents at different ratios of 9:1, 7:1, 5:1, 3:1 and 1:1 were used as gradient eluents for silica gel column chromatography, and samples collected were further isolated and purified by preparative high performance liquid chromatography. Their structures were elucidated by physico-chemical constants and spectroscopic methods. RESULTS: Eight compounds were obtained and identified as 22(Z)-ergosterol-4,6,8,22-tetraene-3-one(1), (22E,24R)-stigma-1,4-diene-3-one(2), (E)-ethyl octadecanoic-16-enoic acid ethyl ester(3), oleanolic acid(4), emodin(5), resveratrol(6), trans-resveratrol glycoside(7) and 6'-gallic acid-4-O-D-resveratrol ester(8).CONCLUSION: Compound 1 is a new natural compound (patent No. ZL 2013 10205435.5), compound 2 and 3 are isolated from this plant for the first time.
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Objective To understand the distribution of Oncomelania hupensis snails and changes of water levels in Gaoyou sections of the Li Canal following the operation of the eastern route project of the South-to-North Water Diversion Project. Methods The Oncomelania snails were monitored in the river banks and water bodies of Gaoyou sections of the Li Canal by means of systematic sampling combined with environmental sampling as well as collection of the floaters from 2014 to 2019, and the water levels were collected in Gaoyou sections of the Li Canal at the typical hydrological year before the operation of the eastern route project of the South-to-North Water Diversion Project and during the period between 2016 and 2019. Results A total area of 235.42 hm2 were investigated and a total of 75.8 kg floaters were collected in Gaoyou sections of the Li Canal from 2014 to 2019; however, no snails were found. The water level in Gaoyou sections of the Li Canal was predominantly high in the flood season and low in the dry season before the operation of the eastern route project of the South-to-North Water Diversion Project, and the water level was elevated in the dry season and relatively low in the flood season after the operation of the project. Conclusion Following the operation of the eastern route project of the South-to-North Water Diversion Project, the original river bank that is characterized by “land in winter and water in summer” has changed in Gaoyou sections of the Li Canal, which is not favorable for snail breeding.
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Objective To compare the effectiveness of snail control between immersion of molluscicides through tide diversion and mollusciciding by spraying in marshland areas. Methods Immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion and spraying 26% suspension concentrate of metaldehyde and niclosamide alone were employed for snail control in two neighboring snail-breeding marshlands, and snails were surveyed before and after mollusciciding. The mortality of snails and the density of living snails were estimated. Results The density of living snails reduced by 72.19% and 100.00% 1 and 2 years after immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion, and 5.93% and 18.15% 1 and 2 years after spraying 26% suspension concentrate of metaldehyde and niclosamide alone. Conclusion Immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion is significantly superior to spraying 26% suspension concentrate of metaldehyde and niclosamide along for snail control, and implementation of immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion for more than 2 successive years may achieve a higher snail control efficiency.
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Objective To evaluate the effect and cost-effectiveness of three commonly used molluscicides, 4% "Luo-wei" (tea-seed distilled saponins, TDS), 50% niclosamide ethanolamine salt wettable powder (NESWP), and 26% metaldehyde and niclosamide suspension concentrate (MNSC) in large-scale field application, so as to provide the references for formulating the strategy of snail control. Methods The field test and parallel comparison were implemented. A marshland with Oncomelania hupensis snails of the Yangtze River was divided into 4 parts (10 hm2) for the research, and three of them were experimental areas while the last one was a blank control area. The experimental areas were sprayed with 4% "Luo-wei", 50% NESWP and 26% MNSC respectively for 3 times and the interval was 1 week. Seven days after each spraying the effect of snail control was investigated, and the costs of molluscicides, labor, transportation, fuel consumption and mechanical loss were recorded. The cost of each molluscicide, snail mortality, snail density, and the cost of increasing 1% of snail mortality per 100 m2 were analyzed and compared. Results After the first, second and third spraying, the corrected snail mortality rates were 67.34%, 76.55% and 84.60% respectively in the 4% "Luo-wei" group; the corrected snail mortality rates were 64.71%, 75.17% and 83.89% respectively in the 50% NESWP group; the corrected snail mortality rates were 66.55%, 76.27% and 86.67% respectively in the 26% MNSC group. There was no significant difference among the 3 groups in the snail mortality at the same spraying time (χ2 = 1.590, 0.571, 3.238, all P > 0.05) . In addition, along with the increase of the spraying times, the snail mortality of each group was increased significantly compared to that of the control group (χ2 = 79.333, 94.718, 117.020, all P < 0.01) . After the first, second and third spraying, the reduction rates of snail density were 69.82%–86.60% in the 4% "Luo-wei" group, 68.66%–86.55% in the 50% NESWP group, and 71.89%–88.87% in the 26% MNSC group respectively. The decreasing amplitude of the snail density was more than 85% in all the experimental areas after 3 rounds of spraying molluscicide. The snail control costs per 100 hm2 were 19.57, 11.97 Yuan and 10.47 Yuan in the 4% "Luo-wei" group, 50% NESWP group, and 26% MNSC group respectively. After the first, second and third spraying, the costs of increasing 1% of snail mortality per 100 m2 were 0.30, 2.08 Yuan and 2.38 Yuan in the 4% "Luo-wei" group, 0.20, 1.10 Yuan and 1.32 Yuan in the 50% NESWP group, and 0.17, 1.04 Yuan and 0.97 Yuan in the 26% MNSC group respectively, and the cost-effectiveness was the highest at the first spraying in all the three groups. Conclusions The effects of the three molluscicides for snail control are similar, but the efficacy of snail control is reduced as the spraying time increases.
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Objective To evaluate the actual effect of the schistosomiasis control program in Jiangsu Province from 2010 to 2015. Methods A total of 67 schistosomiasis-endemic counties in 10 cities were selected, and a combination of retrospective investigation and on-site investigation was adopted to collect and record the epidemic data of the counties from 2010 to 2015, and a retrospective survey database of epidemic situation was established. The effects of integrated control strategies with both Oncomelania hupensis snail control and infection source control were evaluated. Results From 2010 to 2015, 2 465 911 persons who lived in endemic areas were detected for schistosomiasis, with 16 974 positive cases of blood examinations, and 8 positive cases of fecal examinations. Totally 5 145 people with advanced schistosomiasis were treated and 40 460 people with the history of schistosome cercarial-infested water contact received the expanded chemotherapy. A total of 127 636 cattle raised in the endemic areas were detected, and 51 619 cattle (head-times) with the history of cercarial-infested water contact also received the expanded chemotherapy. The area with snails control by molluscicides was 18 604.84 hm2. By the end of 2015, schistosomeinfected snails had not been found and there was no zoological schistosome infection for 5 consecutive years, and in addition, there had been no acute schistosome-infected persons for 6 consecutive years in the whole province. The area with snails dropped to 1 977.18 hm2, with a decreasing rate of 55.24% compared with that in 2010. Conclusion After the implementation of the plan for the prevention and control of schistosomiasis in Jiangsu Province (2010–2015), the prevention and control of schistosomiasis has achieved remarkable effects and realized the goal of the plan.
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Objective To compare the effectiveness of snail control between immersion of molluscicides through tide diversion and mollusciciding by spraying in marshland areas. Methods Immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion and spraying 26% suspension concentrate of metaldehyde and niclosamide alone were employed for snail control in two neighboring snail-breeding marshlands, and snails were surveyed before and after mollusciciding. The mortality of snails and the density of living snails were estimated. Results The density of living snails reduced by 72.19% and 100.00% 1 and 2 years after immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion, and 5.93% and 18.15% 1 and 2 years after spraying 26% suspension concentrate of metaldehyde and niclosamide alone. Conclusion Immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion is significantly superior to spraying 26% suspension concentrate of metaldehyde and niclosamide along for snail control, and implementation of immersion of 26% suspension concentrate of metaldehyde and niclosamide through tide diversion for more than 2 successive years may achieve a higher snail control efficiency.
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A bioassay method for inhibiting platelet aggregation in vitro was established to quantify the pharmacological effects of Compound Danshen Tablets and support its quality control. The inhibition of platelet aggregation in rabbit plasma in vitro by Compound Danshen Tablets was used as the experimental system. The titer was calculated by using the method of dose-response parallel lines. As a result, a bioassay for the inhibition of platelet aggregation in vitro by Compound Danshen Tablets was established. Linearity was good in the concentration range of 0.128 g·mL-1 to 0.205 g·mL-1, and the titer of standard Compound Danshen tablets was 7 659 U·g-1 according to the titer definition. This in vitro assay was simple, reliable, reproducible and convenient. The activity of Compound Danshen Tablets in inhibiting platelet aggregation was quantified by potency assay and the quality of different batches was evaluated. The method can be applied for the quality control of Compound Danshen Tablets.
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The purpose of current study is to investigate the metabolic profile of a triptolide derivative (5R)-5-hydroxytriptolide in vitro. (5R)-5-Hydroxytriptolide was incubated with the hepatocytes of human, monkey, dog, rat or mouse, respectively. Compared with inactivated hepatocytes, four metabolites were identified in hepatocytes from all five species: oxidative ring-opening metabolite (M1), glutathione-conjugating metabolite (M2), and monooxidative combined with glutathione-conjugating metabolites (M3-1 and M3-2), respectively. In human or rat liver microsomes, seven metabolites of (5R)-5-hydroxytriptolide were found, dehydrogenated metabolite (M4) and monooxidative metabolites (M5-1–M5-6), respectively. Reference standards for the metabolites were obtained either through chemical semisynthesis or biotransformation through rat primary hepatocytes. The structures of five metabolites were confirmed, which were 12,13-epoxy ring-opening metabolite M1, 12-glutathione-conjugating metabolite M2, (16S)-, (2R)- and (19R)-monohydroxylated metabolites M5-1, M5-4, and M5-5, respectively. In vitro activity assay revealed that only (2R)-hydroxylated metabolite exhibited weak immunosuppressive activity with less than one-tenth the activity of its parent drug, and a significant decrease in toxicity was observed. It is suggested that (5R)-5-hydroxytriptolide might undergo metabolic inactivation and detoxification in vivo.
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Inflammatory bowel disease(IBD) is a non-specific and chronic recurrent autoimmune disease that involves the gastrointestinal tract. Clinical symptoms of intestinal bleeding, diarrhea, and weight loss threat to human health and induce colorectal cancer. The pathogenesis included living environment, genetic factors, immune cell infiltration and immune stress, weakened mucosal barrier defense and intestinal flora imbalance. At present, clinical treatment drugs mainly include aminosalicylic acid, corticosteroids, immunosuppressants, biological agents, etc., in view of the disadvantages of poor therapeutic effect and expensive price. The active ingredients of traditional Chinese medicine(TCM) in the treatment IBD have various biological activities and multiple targets such as anti-inflammatory, antibacterial, anti-tumor and immune regulation. This article summarized the application and the research progress in protecting intestinal epithelial barrier, maintaining intestinal microbial homeostasis, inhibiting causative factors, and regulating Th1/Th17/Treg balance about TCM in the treatment of IBD. The review provided new ideas for further development of the new drugs on the mechanism based on active ingredients of TCM in IBD treatment.
Subject(s)
Humans , Inflammatory Bowel Diseases , Therapeutics , Intestinal Mucosa , Medicine, Chinese TraditionalABSTRACT
Objective To evaluate the effect and cost-effectiveness of three commonly used molluscicides, 4% "Luo-wei" (tea-seed distilled saponins, TDS), 50% niclosamide ethanolamine salt wettable powder (NESWP), and 26% metaldehyde and niclosamide suspension concentrate (MNSC) in large-scale field application, so as to provide the references for formulating the strategy of snail control. Methods The field test and parallel comparison were implemented. A marshland with Oncomelania hupensis snails of the Yangtze River was divided into 4 parts (10 hm2) for the research, and three of them were experimental areas while the last one was a blank control area. The experimental areas were sprayed with 4% "Luo-wei", 50% NESWP and 26% MNSC respectively for 3 times and the interval was 1 week. Seven days after each spraying the effect of snail control was investigated, and the costs of molluscicides, labor, transportation, fuel consumption and mechanical loss were recorded. The cost of each molluscicide, snail mortality, snail density, and the cost of increasing 1% of snail mortality per 100 m2 were analyzed and compared. Results After the first, second and third spraying, the corrected snail mortality rates were 67.34%, 76.55% and 84.60% respectively in the 4% "Luo-wei" group; the corrected snail mortality rates were 64.71%, 75.17% and 83.89% respectively in the 50% NESWP group; the corrected snail mortality rates were 66.55%, 76.27% and 86.67% respectively in the 26% MNSC group. There was no significant difference among the 3 groups in the snail mortality at the same spraying time (χ2 = 1.590, 0.571, 3.238, all P > 0.05) . In addition, along with the increase of the spraying times, the snail mortality of each group was increased significantly compared to that of the control group (χ2 = 79.333, 94.718, 117.020, all P < 0.01) . After the first, second and third spraying, the reduction rates of snail density were 69.82%–86.60% in the 4% "Luo-wei" group, 68.66%–86.55% in the 50% NESWP group, and 71.89%–88.87% in the 26% MNSC group respectively. The decreasing amplitude of the snail density was more than 85% in all the experimental areas after 3 rounds of spraying molluscicide. The snail control costs per 100 hm2 were 19.57, 11.97 Yuan and 10.47 Yuan in the 4% "Luo-wei" group, 50% NESWP group, and 26% MNSC group respectively. After the first, second and third spraying, the costs of increasing 1% of snail mortality per 100 m2 were 0.30, 2.08 Yuan and 2.38 Yuan in the 4% "Luo-wei" group, 0.20, 1.10 Yuan and 1.32 Yuan in the 50% NESWP group, and 0.17, 1.04 Yuan and 0.97 Yuan in the 26% MNSC group respectively, and the cost-effectiveness was the highest at the first spraying in all the three groups. Conclusions The effects of the three molluscicides for snail control are similar, but the efficacy of snail control is reduced as the spraying time increases.
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Objective To evaluate the actual effect of the schistosomiasis control program in Jiangsu Province from 2010 to 2015. Methods A total of 67 schistosomiasis-endemic counties in 10 cities were selected, and a combination of retrospective investigation and on-site investigation was adopted to collect and record the epidemic data of the counties from 2010 to 2015, and a retrospective survey database of epidemic situation was established. The effects of integrated control strategies with both Oncomelania hupensis snail control and infection source control were evaluated. Results From 2010 to 2015, 2 465 911 persons who lived in endemic areas were detected for schistosomiasis, with 16 974 positive cases of blood examinations, and 8 positive cases of fecal examinations. Totally 5 145 people with advanced schistosomiasis were treated and 40 460 people with the history of schistosome cercarial-infested water contact received the expanded chemotherapy. A total of 127 636 cattle raised in the endemic areas were detected, and 51 619 cattle (head-times) with the history of cercarial-infested water contact also received the expanded chemotherapy. The area with snails control by molluscicides was 18 604.84 hm2. By the end of 2015, schistosomeinfected snails had not been found and there was no zoological schistosome infection for 5 consecutive years, and in addition, there had been no acute schistosome-infected persons for 6 consecutive years in the whole province. The area with snails dropped to 1 977.18 hm2, with a decreasing rate of 55.24% compared with that in 2010. Conclusion After the implementation of the plan for the prevention and control of schistosomiasis in Jiangsu Province (2010–2015), the prevention and control of schistosomiasis has achieved remarkable effects and realized the goal of the plan.
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<p><b>OBJECTIVE</b>To determine the effect of medicated serum of Chinese herbal compound Naofucong (, NFC) on the microglia BV-2 cells viability and the transcription and expression of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in microglia BV-2 cells to further explore the mechanisms underlying the protective effect of NFC on inflammatory process induced by high glucose.</p><p><b>METHODS</b>The microglia BV-2 cells incubated in vitro were divided into different groups: the control group (25 mmol/L glucose), the model group (75 mmol/L glucose), high glucose media containing different dose medicated serum of NFC. After being cultured for 24 h, changes in IL-6 and TNF-α were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The expression of surface marker CD11b of activated microglia was measured by confocal laser scanning microscope and Western blot. Nuclear factor-κB (NF-κB) p-p65 expression was analyzed by Western blot.</p><p><b>RESULTS</b>The model group obviously increased the expression of microglial surface marker CD11b and NF-κB p-p65 (all P<0.01), induced a signifificant up-regulation of release and the mRNA expression of IL-6 and TNF-α (P<0.01 or P<0.05). The medicated serum of NFC could obviously down-regulate the transcription and expression of surface marker CD11 b and NF-κB p-p65 (all P<0.01), and inhibit the mRNA and protein expression (P<0.01 or P<0.05) of inflflammatory cytokines, such as IL-6 and TNF-α, in microglia BV-2 cells cultured with high glucose for 24 h.</p><p><b>CONCLUSIONS</b>The inhibition of microglial activation and IL-6 and TNF-α expression induced by high glucose may at least partly explain NFC therapeutic effects on diabetes-associated cognitive decline diseases. Its underlying mechanism could probably be related to the inhibition of NFC on NF-κB phosphorylation.</p>
Subject(s)
Animals , Male , Mice , Biomarkers , Metabolism , Blotting, Western , CD11b Antigen , Genetics , Metabolism , Cell Line , Cell Shape , Cell Survival , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Glucose , Toxicity , Inflammation , Drug Therapy , Pathology , Interleukin-6 , Genetics , Metabolism , Microscopy, Confocal , RNA, Messenger , Genetics , Metabolism , Rats, Wistar , Real-Time Polymerase Chain Reaction , Transcription Factor RelA , Metabolism , Tumor Necrosis Factor-alpha , Genetics , MetabolismABSTRACT
ABSTARCT:Objective To investigate the effects of the transcription factor SRY-related high-mobility-group box 2 (Sox2)related to stem cells on the invasion and migration of cervical squamous carcinoma cell line SiHa and its mechanism.Methods The expression of Sox2 was detected in Sox2 stably over-expressed cell line SiHa-Sox2 and negative control SiHa-EGFP cells by RT-PCR and Western blotting,respectively.The effects of Sox2 on the invasion and migration capacities of SiHa cells were detected by wound-healing assay and transwell assay.The expression of β-catenin was detected by Western blot.Results Compared with that of SiHa-EGFP cells,the expression of Sox2 at both mRNA and protein levels was obviously upregulated in SiHa-Sox2 cells.The migration and invasion capacities of SiHa-Sox2 cells were increased significantly (P <0.01 ),and the expression of β-catenin increased dramatically compared with that of the control cells.Conclusion Sox2 promotes the invasion and migration capacities of SiHa cells by increasing the expression of β-catenin and activating Wnt signaling pathway, which contributes to the development of cervical cancer.
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@#Objective To study the effects of Guhong injection on the expression of vascular endothelial growth factor (VEGF) in the cortex 14 days after cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were divided into sham group (n=6), ischemia group (n=6), aceglutamide injection group (n=6), Honghua injection group (n=6) and Guhong injection group (n=6). The middle cerebral arteries of all the rats were occluded for 2 hours and reperfusion, except the sham group. Drugs were administered once a day 24 hours after reperfusion. The expression of VEGF in cortex was detected with enzyme-linked immunosorbent assay (ELISA) 14 days after reperfusion. Results The expression of VEGF decreased in the ischemia group compared with the sham group (P<0.001), and it increased both in the aceglutamide and Guhong injection groups compared with the ischemia group (P<0.05). Conclusion Guhong injection can significantly increase the expression of VEGF in the cortex 14 days after ischemia-reperfusion, which may be one of the ways for neuro-protection.
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<p><b>OBJECTIVE</b>To establish the in vitro model of PGE2 released by hypothalamic neurocytes under rrIL-1beta in vitro interference, and investigate the correlation of the PGE2 content and the effect of the drug effect concentration in the model under the effect of Bupleurum injection.</p><p><b>METHOD</b>Hypothalamic neurocytes were cultured in vitro, and added with rrIL-1beta (40 microg x L(-1)) stimulation. Cell sap was collected at different time points. ELISA was adopted to determine the content of PGE2 in cell sap collected at different time points. Hypothalamic neurocytes were cultured in vitro, added with rrIL-1beta (40 microg x L(-1)) stimulation and then different concentrations of Bupleurum injection. The changes in the content of PGE2 in cell supernatant were detected by ELISA. An analysis was made on the linear relationship between the sample concentration and the inhibition rate of PGE2.</p><p><b>RESULT</b>The rrIL-1 cells could stimulate in vitro cultured hypothalamic neurocytes to release PGE2 and reach the peak at 10 h. Bupleurum injection could significantly interfere the release of PGE2 in the in vitro model (P < 0.01, P < 0.05), with a certain linear relationship between the interference effect and the effect concentration of Bupleurum injection (r = 0.911, P < 0.01).</p><p><b>CONCLUSION</b>The rrIL-1 cells could stimulate in vitro cultured hypothalamic neurocytes to release PGE2, with a good correlation between the inhibition and generation effects of PGE2 and the drug concentration.</p>
Subject(s)
Animals , Female , Rats , Biological Assay , Bupleurum , Chemistry , Cells, Cultured , Dinoprostone , Metabolism , Drugs, Chinese Herbal , Pharmacology , Hypothalamus , Cell Biology , Metabolism , Neurons , Metabolism , Rats, Sprague-DawleyABSTRACT
<p><b>BACKGROUND</b>There are two major pathological hallmarks of Alzheimer's disease. One is the progressive accumulation of beta-amyloid (Aβ) in the form of senile plaques; the other is hyperphosphorylated tau, causing neuronal apoptosis. Some inhalation anesthetics, such as isoflurane and desflurane, have been suggested to induce Aβ accumulation and cause AD-like neuropathogenesis. Whether intravenous anesthetics have similar effects is still unclear. We therefore set out to determine the relationship between propofol and AD-like pathogenesis.</p><p><b>METHODS</b>PC12 cells were cultured in serum-free medium for 12 hours prior to drug treatment. Various concentrations from 5 µmol/L to 80 µmol/L of aggregated Aβ25-35 were added to determine a proper concentration for further study. After exposure to 10 µmol/L Aβ25-35 alone or with 20 µmol/L propofol for 6 hours, PC12 cell viability was determined by MTT assay. Western blotting and immunocytochemical staining were performed to observe the protein expression of the Bcl-2 family, tau phosphorylation at different sites, and tau protein kinases and phosphatases.</p><p><b>RESULTS</b>Aβ25-35 induced a decrease in PC12 cell viability in a dose-dependent manner. Exposure to 10 µmol/L Aβ25-35 for 6 hours resulted in the mild cell survival, accompanied by a decline in Bcl-2, and an increase in phosphorylation of GSK-3β and tau at different sites. Compared with the Aβ25-35 group, cells treated with propofol alone showed no significant difference, while cells co-incubated with propofol and Aβ25-35 showed a significantly higher survival rate (P < 0.01 or P < 0.05). Tau phosphorylation at Ser396, Ser404 and Thr231 and the level of GSK-3β in PC12 cells increased after exposure to 10 µmol/L Aβ25-35. Co-incubation with propofol attenuated cellular apoptosis by inhibiting tau phosphorylation.</p><p><b>CONCLUSIONS</b>These data indicate that propofol may protect PC12 cells from Aβ25-35-induced apoptosis and tau hyperphosphorylation through the GSK-3β pathway, therefore it may be a safer anesthesia for AD and elderly patients.</p>
Subject(s)
Animals , Rats , Amyloid beta-Peptides , Pharmacology , Apoptosis , Cell Survival , Glycogen Synthase Kinase 3 , Metabolism , Glycogen Synthase Kinase 3 beta , PC12 Cells , Peptide Fragments , Pharmacology , Phosphorylation , Propofol , Pharmacology , Signal TransductionABSTRACT
Over-expression of CD151 was found to be associated with metastasis and poor prognosis of prostatic carcinoma. This study was designed to examine the mechanism by which CD151 promotes the proliferation and migration of prostatic cancer cells. The pAAV-CD151, pAAV-GFP and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect PC3 cells (a prostatic carcinoma 3 cell line) by the mediation of Fugene HD. Then, the cells were assigned to control group, pAAV-GFP group, pAAV-CD151 group, and pAAV-CD151-AAA group respectively. Cell proliferation was evaluated by using the 3-[4,5-dimet-hylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method. Cell migration assay was performed by using Boyden chambers. The formation of CD151-integrin α3/α6 complex was determined by the method of co-immunoprecipitation. The protein expression levels of CD151 and extracellular signal-regulated kinase (ERK) were measured by Western blotting. The results showed that transfection of pAAV-CD151 or pAAV-CD151-AAA mutant increased the expression of CD151 protein in PC3 cells. Co-immunoprecipitation showed that more CD151-integrin α3/α6 complex was formed in the pAAV-CD151 group than in the control group, the pAAV-GFP group and the pAAV-CD151-AAA mutant group. Furthermore, the proliferative and migrating capacity of PC3 cells was substantially increased in the pAAV-CD151 group but inhibited in the pAAV-CD151-AAA mutant group. CD151 transfection increased the expression of phospho-ERK. Taken together, it was concluded that CD151 promotes the proliferation and migration of PC3 cells through the formation of CD151-integrin complex and the activation of phosphorylated ERK.
Subject(s)
Humans , Male , Cell Line, Tumor , Cell Movement , Cell Proliferation , Integrin alpha3 , Metabolism , Integrin alpha6 , Metabolism , Prostatic Neoplasms , Metabolism , Pathology , Tetraspanin 24 , MetabolismABSTRACT
Over-expression of CD151 was found to be associated with metastasis and poor prognosis of prostatic carcinoma. This study was designed to examine the mechanism by which CD151 promotes the proliferation and migration of prostatic cancer cells. The pAAV-CD151, pAAV-GFP and pAAV-CD151-AAA mutant plasmids were constructed and used to transiently transfect PC3 cells (a prostatic carcinoma 3 cell line) by the mediation of Fugene HD. Then, the cells were assigned to control group, pAAV-GFP group, pAAV-CD151 group, and pAAV-CD151-AAA group respectively. Cell proliferation was evaluated by using the 3-[4,5-dimet-hylthiazol-2-yl]-2,5, diphenyltetrazolium bromide (MTT) method. Cell migration assay was performed by using Boyden chambers. The formation of CD151-integrin α3/α6 complex was determined by the method of co-immunoprecipitation. The protein expression levels of CD151 and extracellular signal-regulated kinase (ERK) were measured by Western blotting. The results showed that transfection of pAAV-CD151 or pAAV-CD151-AAA mutant increased the expression of CD151 protein in PC3 cells. Co-immunoprecipitation showed that more CD151-integrin α3/α6 complex was formed in the pAAV-CD151 group than in the control group, the pAAV-GFP group and the pAAV-CD151-AAA mutant group. Furthermore, the proliferative and migrating capacity of PC3 cells was substantially increased in the pAAV-CD151 group but inhibited in the pAAV-CD151-AAA mutant group. CD151 transfection increased the expression of phospho-ERK. Taken together, it was concluded that CD151 promotes the proliferation and migration of PC3 cells through the formation of CD151-integrin complex and the activation of phosphorylated ERK.
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<p><b>BACKGROUND</b>Gabapentin has been widely and successfully used in the clinic for many neuropathic pain syndromes since last decade, however its analgesic mechanisms are still elusive. Our study was to investigate whether Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) contributes to the analgesic effect of gabapentin on a chronic constriction injury (CCI) model.</p><p><b>METHODS</b>Gabapentin (2%, 100 mg/kg) or saline (0.5 ml/100 g) was injected intraperitoneally 15 minutes prior to surgery and then every 12 hours from postoperative day 0 - 4 to all rats in control, sham and CCI groups. The analgesic effect of gabapentin was assessed by measuring mechanical allodynia and thermal hyperalgesia of rats. Expression and activation of CaMKII were quantified by reverse-transcriptional polymerase chain reaction and Western blotting.</p><p><b>RESULTS</b>The analgesic effect of gabapentin on mechanical allodynia and thermal hyperalgesia was significant in the CCI model, with maximal reduction reached on postoperative day 8. Gabapentin decreased the expression of the total CaMKII and phosphorylated CaMKII in CCI rats.</p><p><b>CONCLUSION</b>The analgesic effect of gabapentin on CCI rats may be related to the decreased expression and phosphorylation of CaMKII in the spinal cord.</p>
Subject(s)
Animals , Male , Rats , Amines , Therapeutic Uses , Analgesics , Therapeutic Uses , Blotting, Western , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Metabolism , Cyclohexanecarboxylic Acids , Therapeutic Uses , Neuralgia , Drug Therapy , Metabolism , Rats, Sprague-Dawley , gamma-Aminobutyric Acid , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of genetic polymorphism in NF-E2-related factor-2 (nrf2) gene promoter locus at 336 in alcoholic liver disease (ALD) with Vibrio vulnificus (VV) sepsis.</p><p><b>METHODS</b>Through the simple random sampling method, C57B6 male mice were divided into normal feeding group (group A, 10 mice), alcoholic liver disease group (group B, 10 mice), normal feeding group infected with VV through intraperitoneal injection (group C, 8 mice), alcoholic liver disease group infected with VV (group D, 110 mice). Through gene sequencing method, nrf2 gene promoter 336 polymorphism in D group was analyzed and grouped into: non-mutation group (336T) (group D1, 7 mice) and mutation group (336C) (group D2, 10 mice). Through RT-PCR, Western-blotting and ELISA method, expressions of nrf2, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), high mobility group protein 1 (HMGB(1)) gene and protein of liver were measured. The pathological changes in liver were recorded with light microscope.</p><p><b>RESULTS</b>After infected with VV for 48 hours for A, B, C, D1, D2 group, the expression medians of nrf2 mRNA in liver were 0.115, 0.173, 0.211, 0.764, 0.352, respectively (χ(2) = 40.64, P < 0.05), the expression medians of IL-10 mRNA in liver were 0.338, 0.637, 1.002, 1.825, 1.403, respectively (χ(2) = 41.05, P < 0.05), the expression medians of TNF-α mRNA in liver were 0.140, 0.254, 0.372, 0.399, 0.699, respectively (χ(2) = 38.16, P < 0.05), the expression medians of HMGB(1) mRNA in liver were 0.230, 0.410, 0.668, 0.508, 1.021, respectively (χ(2) = 31.45, P < 0.05). After infected with VV 48 hours for mice in A, B, C, D1, D2 group, the expression medians of nrf2 protein in liver were 0.908, 1.461, 2.061, 3.982, 2.243, respectively (χ(2) = 33.72, P < 0.05), the expression medians of IL-10 protein in liver were 13.97, 22.54, 30.14, 57.98, 41.53, respectively (χ(2) = 37.31, P < 0.05), the expression medians of TNF-α protein in liver were 114.07, 142.94, 175.44, 174.60, 266.11, respectively (χ(2) = 32.29, P < 0.05), the expression medians of HMGB(1) protein in liver were 2.01, 6.05, 9.62, 6.24, 12.89, respectively (χ(2) = 36.94, P < 0.05). Compared with group A, there were large amount of fat drops, fatty changes in group B, inflammatory cell infiltration, disorder of hepatic cell in group C, and extension of hepatic duct and vein, edema of liver cells and disorder of hepatic cells in group D.</p><p><b>CONCLUSION</b>The nrf2 gene promoter of T336C mutation in C57B6 mouse of ALD can significantly decrease the expression of nrf2, and intensify organ inflammation and damage when they were infected by VV.</p>