Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo ag...Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo against Aβ. Our previous study found that hyperoside suppressed Aβ1-42-induced leakage of the BBB, however, the mechanism remains unclear. In this study, bEnd.3 cells were pretreated with 50, 200, or 500 μM hyperoside for 2 hours, and then exposed to Aβ1-42 for 24 hours. Cell viability was determined using 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling assay were used to analyze cell apoptosis. Western blot assay was carried out to analyze expression levels of Bax, Bcl-2, cytochrome c, caspase-3, caspse-8, caspase-9, caspase-12, occludin, claudin-5, zonula occludens-1, matrix metalloproteinase-2（MMP-2）, and MMP-9. Exposure to Aβ1-42 alone remarkably induced bEnd.3 cell apoptosis; increased ratios of cleaved caspase-9/caspase-9, Bax/Bcl-2, cleav ed caspase-8/caspase-8, and cleaved caspase-12/caspase-12; increased expression of cytochrome c and activity of caspase-3; diminished levels of zonula occludens-1, claudin-5, and occludin; and increased levels of MMP-2 and MMP-9. However, hyperoside pretreatment reversed these changes in a dose-dependent manner. Our findings confirm that hyperoside alleviates fibrillar Aβ1-42-induced BBB disruption, thus offering a feasible therapeutic application in Alzheimer＇s disease.展开更多
OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aim...OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma.METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta.neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index(CI).Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis.The expression of apoptosis-related proteins were detected by Western blot.RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50 of 5.6 μmol·L^(-1),and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation,with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth.Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control,GRN A,Cisplatin,and combination of GRN A and Cisplatin groups,respectively.Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.展开更多
OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells...OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms.METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1).The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice.The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy(TEM) andflow cytometry.Autophagy related proteins were detected by Western blotting.RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi.ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice.TEM and flow cytometry showed that Zn-CuO NPS induced autophagy,and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting.CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.展开更多
CRISPR/Cas9-mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus(PRV) could escape from CRISPR/Cas9...CRISPR/Cas9-mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus(PRV) could escape from CRISPR/Cas9-mediated inhibition.In order to elucidate whether the escape of PRV from Cas9-mediated inhibition was due to cell deficiencies, such as genetic instability of sgRNA or Cas9 protein, the positive cells were passaged ten times, and PRV infection in the sgRNA-expressing cells was evaluated in the present study. The results showed that subculturing cells has no effect on Cas9-mediated cleavage of PRV. Different passages of PX459-PRV cells can stably express sgRNA to facilitate Cas9/sgRNA cleavage on the UL30 gene of PRV, resulting in a pronounced inhibition of PRV infection. Studies to elucidate the mechanism of PRV escape are currently in progress.展开更多
Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissecte...Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells （DAC-CM）. Immunocytochemistry, reverse transcriptase polymerase chain reaction （RT-PCR） and scanning electron microscope （SEM） were performed to compare the biolo- gical change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex （DAC） condi- tioned medium, we found the microenvironment created by root could abrogate the ＂crown＂ features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.展开更多
PCV2 is considered the main pathogen of porcine circovirus diseases and porcine circovirus-associated diseases(PCVD/PCVAD). However, the exact mechanism underlying PCVD/PCVAD is currently unknown. Mouse models of PCV2...PCV2 is considered the main pathogen of porcine circovirus diseases and porcine circovirus-associated diseases(PCVD/PCVAD). However, the exact mechanism underlying PCVD/PCVAD is currently unknown. Mouse models of PCV2 are valuable experimental tools that can shed light on the pathogenesis of infection and will enable the evaluation of antiviral agents and vaccine candidates. In this review, we discuss the current state of knowledge of mouse models used in PCV2 research that has been performed to date, highlighting their strengths and limitations, as well as prospects for future PCV2 studies.展开更多
Background To observe the anti-proliferative effects and molecular mechanisms of traditional Chinese medicine prescriptions Xi-Huang pellet （XHP） on human breast cancer cell line SKBR3. Materials and Methods： Huma...Background To observe the anti-proliferative effects and molecular mechanisms of traditional Chinese medicine prescriptions Xi-Huang pellet （XHP） on human breast cancer cell line SKBR3. Materials and Methods： Human breast cancer cell line SKBR3 was cultured. Trypan blue exclusion assay was used to investigate the anti-proliferative effects of medicine. The rates of the cell cycle were measured by ^ow cytometry （FCM）. The impacts of medicine on the protein expression were detected by Enzyme-linked immunosorbent assay （ELISA）. RNA extraction and real-time PCR were used to observe the change of RNA expression. Results： Myrrh, XHP and XHP combined 5-FU could inhibit cellproliferation and arrest SKBR3 cells at S phases. XHP combined 5-FU resulted in a significant increase of intracellular p21WAF1/CiP1 content in SKBR3 cells. XHP and XHP combined 5-FU led to a decrease of mRNA expression on cyclin A2, cyclin D1 and cyclin E in SKBR3 cells. Conclusions： XHP inhibits the proliferation of breast cancer SKBR3 cells via a significantly change of cyclins expression and p21WAF1/CIP1 pathway. 5-FU significantly enhanced the inhibitory effects of XHP on SKBR3 cells.展开更多
AIM:To propose an appropriate staging system for hepatocellular carcinoma(HCC)classification.METHODS:Here,288 in-patients with HCC were studied and divided into three groups:those with expansive growth,invasive growth...AIM:To propose an appropriate staging system for hepatocellular carcinoma(HCC)classification.METHODS:Here,288 in-patients with HCC were studied and divided into three groups:those with expansive growth,invasive growth(including satellite nodules,nodule fusions and direct tumor invasion of adjacent organs),or disseminative growth(including vascular involvement,regional lymph node metastasis and distant metastasis).A survival analysis was performed using a Kaplan-Meier analysis,and prognostic factors for overall survival were determined by the Cox proportional hazards regression model.RESULTS:The overall survival(OS)of patients with invasive tumor growth was shorter than that of patients with expansive tumor growth(27.796±3.730and 57.398±4.873 mo,respectively,P<0.001).No significant difference in survival was observed between patients with vascular involvement and patients with regional lymph node metastasis(21.667±4.773 and14.619±2.456 mo,respectively,P=0.801).The OS of patients with distant metastasis(6.417±1.395mo)was shorter than that of the other groups(P<0.001).No significant difference in survival was observed between patients with expansive tumor growth and vascular and/or regional lymph node involvement and patients with invasive tumor growth and no vascular and/or lymph node involvement(25.762±7.024,21.200±7.794 and 39.533±5.840 mo,respectively;P=0.871,0.307 and 0.563,respectively).CONCLUSION:These data led to the proposal of a new staging system:the Expansive-Invasive-Disseminative growth staging classification.展开更多
OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mec...OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti.tumor effects and mechanisms of water extract of cloves(WEC) against colorectal cancer.METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3 a/b) were determined by western blot.RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50 values of 150 μg·mL-1.The colonyformation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice.Flow cytometry result showed that WEC significantly induced autophagy,and the averaged relative values of fluorescence intensity were206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1 WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I.CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo,which might be related with autophagy induction,and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.展开更多
Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction m...Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction methods,namely,water extraction,ultrasonic extraction,enzymatic hydrolysis-water extraction,and enzymatic hydrolysis-ultrasonic extraction were considered to determine the best extraction method.Single factor and orthogonal experiments were performed to determine the optimum extracting conditions.Sevage,enzymatic hydrolysis-Sevage,trichloroacetic acid(TCA),TCA-Sevage and enzymatic hydrolysis-TCA methods were tested to determine the best deproteinization method.The glycopeptide content and protein removal ratio were analyzed by the phenol-sulfuric acid and Coomassie Brilliant Blue methods.Results:Gekko sulfated glycopeptide obtained by water extraction could inhibit the proliferation of HepG2 and SKBR3,as well as promote that of lymphocytes.The glycopeptide content was 4.049%in the optimal extracting condition of a triple decoction extraction for 1 hour each time with a material to solvent ratio of 1:15.The enzymatic hydrolysis-TCA method was found to be the optimal deproteinization method,with a protein removal ratio of 50.46%,glycopeptide content of 14.27%,and inhibitory ratio on human HepG2 cells of 49.06%.Conclusion:This extraction and purification technique for Gekko sulfated glycopeptide is reasonable,feasible,and provides a scientific basis for industrial production.展开更多
High throughput gene expression analysis is swiftly becoming the focal point for deciphering molecular mechanisms underlying various dif-ferent biological questions. Testament to this is the fact that vast volumes of ...High throughput gene expression analysis is swiftly becoming the focal point for deciphering molecular mechanisms underlying various dif-ferent biological questions. Testament to this is the fact that vast volumes of expression profiles are being generated rapidly by scientists worldwide and subsequently stored in publicly available data repositories such as ArrayEx-press and the Gene Expression Omnibus (GEO). Such wealth of biological data has motivated biologists to compare expression profiles gen-erated from biologically-related microarray ex-periments in order to unravel biological mecha-nisms underlying various states of diseases. However, without the availability of appropriate software and tools, they are compelled to use manual or labour-intensive methods of com-parisons. A scrutiny of current literature makes it apparent that there is a soaring need for such bioinformatics tools that cater for the multiple analyses of expression profiles. In order to contribute towards this need, we have developed an efficient software pipeline for the analysis of multiple gene expression data-sets, called Micropath, which implements three principal functions;1) it searches for common genes amongst n number of datasets using a number crunching method of comparison as well as applying the principle of permutations and combinations in the form of a search strat-egy, 2) it extracts gene expression patterns both graphically and statistically, and 3) it streams co-expressed genes to all molecular pathways belonging to KEGG in a live fashion. We sub-jected MicroPath to several expression datasets generated from our tolerance-related in-house microarray experiments as well as published data and identified a set of 31 candidate genes that were found to be co-expressed across all interesting datasets. Pathway analysis revealed their putative roles in regulating immune toler-ance. MicroPath is freely available to download from: www.1066technologies.co.uk/micropath.展开更多
Recent years,the immune checkpoint blockers(ICBs)become more and more important in tumor therapy with the development of tumor immunology and achieved significant clinical effects in various solid tumors.ICBs is a typ...Recent years,the immune checkpoint blockers(ICBs)become more and more important in tumor therapy with the development of tumor immunology and achieved significant clinical effects in various solid tumors.ICBs is a type of tumor immunotherapy method which controls and clears tumor by activating the autoimmune system and restoring the body's normal anti-tumor immune response.However,the resistance of ICBs cannot be ignored and still faces many challenges.The mechanism and the solution of ICBs resistance need to be further research.展开更多
Objective:Several studies indicated that many factors have relationship with the metastatic sites of advanced breast cancer.This retrospective study investigated the high risk factors which related to the different me...Objective:Several studies indicated that many factors have relationship with the metastatic sites of advanced breast cancer.This retrospective study investigated the high risk factors which related to the different metastatic sites of stage IV breast cancer.Patients and methods:From January 2003 to December 2005 a total of 387 consecutive breast cancer patients were retrospectively analyzed.The relationships between different categorical variables and breast cancer were identified by Chi-square tests.Results:The high risk factors of metastatic breast cancer included the overexpression of HER-2 and lymph nodes invasion.The overexpression of HER-2 and lymph nodes invasion had a significantly difference between metastatic breast cancer and breast cancer without metastasis(P=0.018,P<0.001,respectively).As for metastatic breast cancer patients with only one single metastatic organ,the overexpression of HER-2 had a significantly high positive rate in patients with visceral metastases when compared with bone metastasis(P=0.045).Conclusion:The overexpression of HER-2 and lymph nodes invasion significantly influenced the metastasis of breast cancer.Overexpression of Her-2 was high risk factors for breast cancer developed to visceral metastases disease.展开更多
Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting b...Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting basic fibroblast growth factor(bFGF).bFGF promoted the proliferation of fibroblasts.Both fibrosarcoma and fibroblasts derived from fibrous connective tissue.This study investigated whether GSPP has the inhibitory effects on human fibrosarcoma HT1080 cells.Materials and methods:The trypan blue exclusion assay was used to determine cell viability and cell numbers.Cells migration was observed by wound-healing and transwell.Results:From the first day to seventh day,HT1080 cells number of GSPP,bFGF,GSPP combined bFGF groups had not change compared with control.HT1080 cells migration distance and the number of migrating cells of GSPP,bFGF,GSPP combined bFGF groups were not significantly reduced.Conclusions:GSPP did not have inhibitory effects on the proliferation and migration of human fibrosarcoma HT1080 cells.Thus further research should be carried out in order to study the mechanism of GSPP and bFGF acting on the tumor stroma.展开更多
Tumor microenvironment plays a very important role in the growth,invasion and metastasis of tumor cells.The tumor interstitial microenvironment is an important part of the tumor microenvironment,which includes two par...Tumor microenvironment plays a very important role in the growth,invasion and metastasis of tumor cells.The tumor interstitial microenvironment is an important part of the tumor microenvironment,which includes two parts:the non-cellular and cellular components of the tumor interstitium,specifically including the extracellular matrix,blood vessels,and interstitial cells.Among them,activated interstitial fibroblasts,namely cancer-associated fibroblasts(CAFs),are the main components of tumor interstitial cells,which are most closely related to tumor interstitial fibrosis and tumor progress,and are expected to become a new target for cancer treatment.展开更多
Tumor metastasis is the basic feature and important sign of malignant tumors,and the main cause of death for most cancer patients.The tumor microenvironment is closely related to tumor metastasis and a deep understand...Tumor metastasis is the basic feature and important sign of malignant tumors,and the main cause of death for most cancer patients.The tumor microenvironment is closely related to tumor metastasis and a deep understanding of the relationship between tumor microenvironment and tumor metastasis has positive guiding significance for the prevention and treatment of tumors.The occurrence,development and metastasis of tumors involve a series of complex biological processes.In the process of tumor metastasis,the tumor microenvironment cooperates with tumor cells.Tumor microenvironment is composed of cellular components,physical components and biochemical components,and it participates in important biological processes such as tumor growth,invasion,metastasis,neovascularization,immune escape,and tumor drug resistance.The study of tumor microenvironment can help us better understand the biological behavior of tumor and control the occurrence,development and metastasis of tumor.展开更多
Tumor immunotherapy has been a research hotspot in the field of tumor therapy in recent years,especially the successful development and application of immune checkpoint inhibitors,which has brought tumor immunotherapy...Tumor immunotherapy has been a research hotspot in the field of tumor therapy in recent years,especially the successful development and application of immune checkpoint inhibitors,which has brought tumor immunotherapy into a new era.Unlike conventional treatment methods,such as chemotherapy,radiotherapy,and targeted therapy which directly affect tumor cells,immune checkpoint inhibitors block tumor immune environment checkpoint pathways and stimulate tumor-specific T cell functions to achieve anti-tumor effects.However,the clinical efficacy evaluation of immune checkpoint blockers still faces many challenges,and the solid tumor evaluation criteria(response evaluation criteria in solid tumors,RECIST)applicable to traditional chemotherapy drugs cannot accurately assess the efficacy of immunotherapy.Immune-related response criteria need further research.展开更多
基金financially supported by the National Natural Science Foundation of China,No.81573771the Natural Science Foundation of Jiangsu Province of China,No.BK20151599
文摘Mounting evidence indicates that amyloid β protein（Aβ） exerts neurotoxicity by disrupting the blood-brain barrier（BBB） in Alzheimer＇s disease. Hyperoside has neuroprotective effects both in vitro and in vivo against Aβ. Our previous study found that hyperoside suppressed Aβ1-42-induced leakage of the BBB, however, the mechanism remains unclear. In this study, bEnd.3 cells were pretreated with 50, 200, or 500 μM hyperoside for 2 hours, and then exposed to Aβ1-42 for 24 hours. Cell viability was determined using 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling assay were used to analyze cell apoptosis. Western blot assay was carried out to analyze expression levels of Bax, Bcl-2, cytochrome c, caspase-3, caspse-8, caspase-9, caspase-12, occludin, claudin-5, zonula occludens-1, matrix metalloproteinase-2（MMP-2）, and MMP-9. Exposure to Aβ1-42 alone remarkably induced bEnd.3 cell apoptosis; increased ratios of cleaved caspase-9/caspase-9, Bax/Bcl-2, cleav ed caspase-8/caspase-8, and cleaved caspase-12/caspase-12; increased expression of cytochrome c and activity of caspase-3; diminished levels of zonula occludens-1, claudin-5, and occludin; and increased levels of MMP-2 and MMP-9. However, hyperoside pretreatment reversed these changes in a dose-dependent manner. Our findings confirm that hyperoside alleviates fibrillar Aβ1-42-induced BBB disruption, thus offering a feasible therapeutic application in Alzheimer＇s disease.
文摘OBJECTIVE Granulin A(GRN A),a cytokinesis protein,is derived from proteolysis of progranulin.The previous study in our laboratory has shown that GRN A is able to inhibit cancer cell growth significantly.This study aimed to investigate the effect of combination of GRN A and cisplatin on in vitro and in vivo on the growth of hepatocellular carcinoma.METHODS The in vitro and in vivo antitumor effects of combination of GRN A and Cisplatin were evaluated with MTS assay and subcuta.neous transplantation tumor model.Chou-Talalay method was used to calculate the combination index(CI).Colony formation assay and flow cytometry were used to detect the effects of GRN A on apoptosis.The expression of apoptosis-related proteins were detected by Western blot.RESULTS MTS assay showed that GRN A significantly inhibit hepatocellular carcinoma cells growth with the IC50 of 5.6 μmol·L^(-1),and GRN A combined with cisplatin synergistically inhibit hepatocellular carcinoma proliferation,with the CI<1.The colony-formation assay showed that GRN A significantly enhanced the inhibitory effects of cisplatin on cellular anchorage-independent growth.Flow cytometry showed that GRN A combined with cisplatin synergistically induced apoptosis,with the apoptotic rates of 5.87%,32.74%,35.67% and 67.15% in control,GRN A,Cisplatin,and combination of GRN A and Cisplatin groups,respectively.Western blot confirmed that the two drugs synergistically changed the expressions of proteins related to apoptosis.In vivo experiment indicated that combination of GRN A and cisplatin significantly suppressed tumor growth compared with single drug treatment groups.CONCLUSION The combination of GRN A and cisplatin resulted in synergistic antitumor effects against hepatocellular carcinoma both in vitro and in vivo.
文摘OBJECTIVE Zn-doped CuO nanocomposites(Zn-CuO NPs) are novel nanoparticles synthesized by our research group.In this study,we assessed the in vitro and in vivo antitumor effects of Zn-CuO NPS on pancreatic cancer cells,as well as the potential mechanisms.METHODS MTS assay was used to detect the effects of Zn-CuO NPS on proliferation pancreatic cancer cells(Panc-mia and Aspc-1).The in vivo antitumor effects of Zn-CuO NPs were detected by xenografts model in nude mice.The effects of Zn-CuO NPS on autophagy were detected bytransmission electron microscopy(TEM) andflow cytometry.Autophagy related proteins were detected by Western blotting.RESULTS Zn-CuO NPS significantly inhibited the proliferation of Panc-mia cells and Aspc-1 cells.In vivo experi.ments showed that Zn-CuO NPS significantly inhibited the tumor growth in nude mice without affecting the body weight of the mice.TEM and flow cytometry showed that Zn-CuO NPS induced autophagy,and significantly increased the number of autophagosome.Western Blot showed that Zn-CuO NPS alterd the expression of autophagy related proteins,such as AMPK,mTORand Beclin-1.Also,AMPK inhibitor could significantly reduce Zn-CuO NPS-induced autophagy pathwayas analyzed byWestern blotting.CONCLUSION The findings suggested that Zn-doped CuO nanocomposites inhibited the in vitro and in vivo growth of pancreatic cancer by inducing autophagy through AMPK/mTOR pathway.
基金financially supported by the National Key Research and Development Program of China(No.2017YFD0500103)the Beijing Natural Science Foundation(No.5152023)+4 种基金the National Natural Science Foundation of China(No.31772747 and31272385)the Jilin Province Science and Technology Development Projects(20150204077NY)the Graduate Innovation Fund of Jilin Universitythe Program for Chang jiang Scholarsthe University Innovative Research Team(No.IRT1248)
文摘CRISPR/Cas9-mediated genome editing can inhibit virus infection by targeting the conserved regions of the viral genomic DNA. Unexpectedly, we found previously that pseudorabies virus(PRV) could escape from CRISPR/Cas9-mediated inhibition.In order to elucidate whether the escape of PRV from Cas9-mediated inhibition was due to cell deficiencies, such as genetic instability of sgRNA or Cas9 protein, the positive cells were passaged ten times, and PRV infection in the sgRNA-expressing cells was evaluated in the present study. The results showed that subculturing cells has no effect on Cas9-mediated cleavage of PRV. Different passages of PX459-PRV cells can stably express sgRNA to facilitate Cas9/sgRNA cleavage on the UL30 gene of PRV, resulting in a pronounced inhibition of PRV infection. Studies to elucidate the mechanism of PRV escape are currently in progress.
基金supported by National Nature Science Foundation of China(Project No.3057 2046,30725042)
文摘Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells （DAC-CM）. Immunocytochemistry, reverse transcriptase polymerase chain reaction （RT-PCR） and scanning electron microscope （SEM） were performed to compare the biolo- gical change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex （DAC） condi- tioned medium, we found the microenvironment created by root could abrogate the ＂crown＂ features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.
基金National Key Research and Development Program of China,Grant/Award Number:2017YFD0500103National Natural Science Foundation of China,Grant/Award Number:31772747,31272385+3 种基金Jilin Province Science and Technology Development Projects,Grant/Award Number:20150204077NYGraduate Innovation Fund of Jilin Universitythe Program for Changjiang Scholarsthe University Innovative Research Team,Grant/Award Number:IRT1248
文摘PCV2 is considered the main pathogen of porcine circovirus diseases and porcine circovirus-associated diseases(PCVD/PCVAD). However, the exact mechanism underlying PCVD/PCVAD is currently unknown. Mouse models of PCV2 are valuable experimental tools that can shed light on the pathogenesis of infection and will enable the evaluation of antiviral agents and vaccine candidates. In this review, we discuss the current state of knowledge of mouse models used in PCV2 research that has been performed to date, highlighting their strengths and limitations, as well as prospects for future PCV2 studies.
基金This work was supported by grants from the National Science Foundation of China （No. 81173376） and New Century Excellent Talent （NCET-11-1068）.
文摘Background To observe the anti-proliferative effects and molecular mechanisms of traditional Chinese medicine prescriptions Xi-Huang pellet （XHP） on human breast cancer cell line SKBR3. Materials and Methods： Human breast cancer cell line SKBR3 was cultured. Trypan blue exclusion assay was used to investigate the anti-proliferative effects of medicine. The rates of the cell cycle were measured by ^ow cytometry （FCM）. The impacts of medicine on the protein expression were detected by Enzyme-linked immunosorbent assay （ELISA）. RNA extraction and real-time PCR were used to observe the change of RNA expression. Results： Myrrh, XHP and XHP combined 5-FU could inhibit cellproliferation and arrest SKBR3 cells at S phases. XHP combined 5-FU resulted in a significant increase of intracellular p21WAF1/CiP1 content in SKBR3 cells. XHP and XHP combined 5-FU led to a decrease of mRNA expression on cyclin A2, cyclin D1 and cyclin E in SKBR3 cells. Conclusions： XHP inhibits the proliferation of breast cancer SKBR3 cells via a significantly change of cyclins expression and p21WAF1/CIP1 pathway. 5-FU significantly enhanced the inhibitory effects of XHP on SKBR3 cells.
基金Supported by National Science Foundation of ChinaNo.81173376+1 种基金New Century Excellent TalentNo.NCET11-1068
文摘AIM:To propose an appropriate staging system for hepatocellular carcinoma(HCC)classification.METHODS:Here,288 in-patients with HCC were studied and divided into three groups:those with expansive growth,invasive growth(including satellite nodules,nodule fusions and direct tumor invasion of adjacent organs),or disseminative growth(including vascular involvement,regional lymph node metastasis and distant metastasis).A survival analysis was performed using a Kaplan-Meier analysis,and prognostic factors for overall survival were determined by the Cox proportional hazards regression model.RESULTS:The overall survival(OS)of patients with invasive tumor growth was shorter than that of patients with expansive tumor growth(27.796±3.730and 57.398±4.873 mo,respectively,P<0.001).No significant difference in survival was observed between patients with vascular involvement and patients with regional lymph node metastasis(21.667±4.773 and14.619±2.456 mo,respectively,P=0.801).The OS of patients with distant metastasis(6.417±1.395mo)was shorter than that of the other groups(P<0.001).No significant difference in survival was observed between patients with expansive tumor growth and vascular and/or regional lymph node involvement and patients with invasive tumor growth and no vascular and/or lymph node involvement(25.762±7.024,21.200±7.794 and 39.533±5.840 mo,respectively;P=0.871,0.307 and 0.563,respectively).CONCLUSION:These data led to the proposal of a new staging system:the Expansive-Invasive-Disseminative growth staging classification.
文摘OBJECTIVE Cloves(Syzygium aromaticumL.) have been used as both a spice and a traditional Chinese medicinal herb for thousands of years.However,relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti.tumor effects and mechanisms of water extract of cloves(WEC) against colorectal cancer.METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3 a/b) were determined by western blot.RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50 values of 150 μg·mL-1.The colonyformation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice.Flow cytometry result showed that WEC significantly induced autophagy,and the averaged relative values of fluorescence intensity were206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1 WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I.CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo,which might be related with autophagy induction,and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.
基金grants from the National Science Foundation of China(81173376).
文摘Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide,removal ratio of proteins,and anticancer activities.Methods:Different extraction methods,namely,water extraction,ultrasonic extraction,enzymatic hydrolysis-water extraction,and enzymatic hydrolysis-ultrasonic extraction were considered to determine the best extraction method.Single factor and orthogonal experiments were performed to determine the optimum extracting conditions.Sevage,enzymatic hydrolysis-Sevage,trichloroacetic acid(TCA),TCA-Sevage and enzymatic hydrolysis-TCA methods were tested to determine the best deproteinization method.The glycopeptide content and protein removal ratio were analyzed by the phenol-sulfuric acid and Coomassie Brilliant Blue methods.Results:Gekko sulfated glycopeptide obtained by water extraction could inhibit the proliferation of HepG2 and SKBR3,as well as promote that of lymphocytes.The glycopeptide content was 4.049%in the optimal extracting condition of a triple decoction extraction for 1 hour each time with a material to solvent ratio of 1:15.The enzymatic hydrolysis-TCA method was found to be the optimal deproteinization method,with a protein removal ratio of 50.46%,glycopeptide content of 14.27%,and inhibitory ratio on human HepG2 cells of 49.06%.Conclusion:This extraction and purification technique for Gekko sulfated glycopeptide is reasonable,feasible,and provides a scientific basis for industrial production.
文摘High throughput gene expression analysis is swiftly becoming the focal point for deciphering molecular mechanisms underlying various dif-ferent biological questions. Testament to this is the fact that vast volumes of expression profiles are being generated rapidly by scientists worldwide and subsequently stored in publicly available data repositories such as ArrayEx-press and the Gene Expression Omnibus (GEO). Such wealth of biological data has motivated biologists to compare expression profiles gen-erated from biologically-related microarray ex-periments in order to unravel biological mecha-nisms underlying various states of diseases. However, without the availability of appropriate software and tools, they are compelled to use manual or labour-intensive methods of com-parisons. A scrutiny of current literature makes it apparent that there is a soaring need for such bioinformatics tools that cater for the multiple analyses of expression profiles. In order to contribute towards this need, we have developed an efficient software pipeline for the analysis of multiple gene expression data-sets, called Micropath, which implements three principal functions;1) it searches for common genes amongst n number of datasets using a number crunching method of comparison as well as applying the principle of permutations and combinations in the form of a search strat-egy, 2) it extracts gene expression patterns both graphically and statistically, and 3) it streams co-expressed genes to all molecular pathways belonging to KEGG in a live fashion. We sub-jected MicroPath to several expression datasets generated from our tolerance-related in-house microarray experiments as well as published data and identified a set of 31 candidate genes that were found to be co-expressed across all interesting datasets. Pathway analysis revealed their putative roles in regulating immune toler-ance. MicroPath is freely available to download from: www.1066technologies.co.uk/micropath.
文摘Recent years,the immune checkpoint blockers(ICBs)become more and more important in tumor therapy with the development of tumor immunology and achieved significant clinical effects in various solid tumors.ICBs is a type of tumor immunotherapy method which controls and clears tumor by activating the autoimmune system and restoring the body's normal anti-tumor immune response.However,the resistance of ICBs cannot be ignored and still faces many challenges.The mechanism and the solution of ICBs resistance need to be further research.
文摘Objective:Several studies indicated that many factors have relationship with the metastatic sites of advanced breast cancer.This retrospective study investigated the high risk factors which related to the different metastatic sites of stage IV breast cancer.Patients and methods:From January 2003 to December 2005 a total of 387 consecutive breast cancer patients were retrospectively analyzed.The relationships between different categorical variables and breast cancer were identified by Chi-square tests.Results:The high risk factors of metastatic breast cancer included the overexpression of HER-2 and lymph nodes invasion.The overexpression of HER-2 and lymph nodes invasion had a significantly difference between metastatic breast cancer and breast cancer without metastasis(P=0.018,P<0.001,respectively).As for metastatic breast cancer patients with only one single metastatic organ,the overexpression of HER-2 had a significantly high positive rate in patients with visceral metastases when compared with bone metastasis(P=0.045).Conclusion:The overexpression of HER-2 and lymph nodes invasion significantly influenced the metastasis of breast cancer.Overexpression of Her-2 was high risk factors for breast cancer developed to visceral metastases disease.
基金This work was supported by grants from the New Century Excellent Talent(NCET-11-1068)National Science Foundation of China(No.81173376).
文摘Background:Fibrosarcoma is a malignant soft tissue tumor of mesenchymal origin.Gekko sulfated glycopeptide(GSPP),an anticancer drug in traditional Chinese medicine,could inhibited the tumor angiogenesis by targeting basic fibroblast growth factor(bFGF).bFGF promoted the proliferation of fibroblasts.Both fibrosarcoma and fibroblasts derived from fibrous connective tissue.This study investigated whether GSPP has the inhibitory effects on human fibrosarcoma HT1080 cells.Materials and methods:The trypan blue exclusion assay was used to determine cell viability and cell numbers.Cells migration was observed by wound-healing and transwell.Results:From the first day to seventh day,HT1080 cells number of GSPP,bFGF,GSPP combined bFGF groups had not change compared with control.HT1080 cells migration distance and the number of migrating cells of GSPP,bFGF,GSPP combined bFGF groups were not significantly reduced.Conclusions:GSPP did not have inhibitory effects on the proliferation and migration of human fibrosarcoma HT1080 cells.Thus further research should be carried out in order to study the mechanism of GSPP and bFGF acting on the tumor stroma.
文摘Tumor microenvironment plays a very important role in the growth,invasion and metastasis of tumor cells.The tumor interstitial microenvironment is an important part of the tumor microenvironment,which includes two parts:the non-cellular and cellular components of the tumor interstitium,specifically including the extracellular matrix,blood vessels,and interstitial cells.Among them,activated interstitial fibroblasts,namely cancer-associated fibroblasts(CAFs),are the main components of tumor interstitial cells,which are most closely related to tumor interstitial fibrosis and tumor progress,and are expected to become a new target for cancer treatment.
文摘Tumor metastasis is the basic feature and important sign of malignant tumors,and the main cause of death for most cancer patients.The tumor microenvironment is closely related to tumor metastasis and a deep understanding of the relationship between tumor microenvironment and tumor metastasis has positive guiding significance for the prevention and treatment of tumors.The occurrence,development and metastasis of tumors involve a series of complex biological processes.In the process of tumor metastasis,the tumor microenvironment cooperates with tumor cells.Tumor microenvironment is composed of cellular components,physical components and biochemical components,and it participates in important biological processes such as tumor growth,invasion,metastasis,neovascularization,immune escape,and tumor drug resistance.The study of tumor microenvironment can help us better understand the biological behavior of tumor and control the occurrence,development and metastasis of tumor.
文摘Tumor immunotherapy has been a research hotspot in the field of tumor therapy in recent years,especially the successful development and application of immune checkpoint inhibitors,which has brought tumor immunotherapy into a new era.Unlike conventional treatment methods,such as chemotherapy,radiotherapy,and targeted therapy which directly affect tumor cells,immune checkpoint inhibitors block tumor immune environment checkpoint pathways and stimulate tumor-specific T cell functions to achieve anti-tumor effects.However,the clinical efficacy evaluation of immune checkpoint blockers still faces many challenges,and the solid tumor evaluation criteria(response evaluation criteria in solid tumors,RECIST)applicable to traditional chemotherapy drugs cannot accurately assess the efficacy of immunotherapy.Immune-related response criteria need further research.
