1.Correlation between the VEGF-C and VEGF-D expressions and the retroperitoneal and inguinal lymph node metastasis in human epithelial ovarian carcinoma
Yuzhen WANG ; Zhiqing LIANG ; Ling LONG ; Dan WANG ;
Journal of Third Military Medical University 2003;0(11):-
Objective To investigate the correlation between the expressions of vascular endothelial growth factor C (VEGF C) and vascular endothelial growth factor D (VEGF D) and the lymph node metastasis of groin and retroperitoneum in human epithelial ovarian carcinoma. Methods The expressions of VEGF C and VEGF D in benign, borderline, and malignant epithelial ovarian tumors were detected by immunohistochemistry and light microscopy. Results The expressions of VEGF C and VEGF D were found in benign, borderline, and malignant epithelial ovarian tumors. The expressions of VEGF C and VEGF D in ovarian carcinomas were significantly higher than those in benign and borderline ovarian tumors. The expression levels of VEGF C and VEGF D were correlated with peritoneal metastasis, lymph node metastasis of groin and retroperitoneum, and clinical stage, but not with distant metastasis, histology type, histological grade, and age. Conclusion Up regulation of VEGF C and VEGF D in epithelial ovarian carcinoma is advantageous to lymph node metastasis. This process may probably be correlated with lymphangiogenesis in tumors.
2.Oleanolic acid induces G₂/M phase arrest and apoptosis in human hepatocellular carcinoma Bel-7402 cells.
Ling LIU ; Jian-long ZHAO ; Jian-gang WANG
China Journal of Chinese Materia Medica 2015;40(24):4897-4902
This study was to examine the mechanism of oleanolic acid (OA) induces G2/M phase arrest and apoptosis in human hepatocellular carcinoma Bel-7402 cells. MTT and trypan blue exclusion test assay were adopted to detect the proliferate status of cells treated with OA. We assayed the cell cycle by flow cytometry using PI staining. Apoptosis was determined by Annexin V-FITC staining and PI labeling. The expressions of cycle related proteins and apoptotic related proteins were determined by Western blot analysis. OA strongly inhibited human hepatoma cells proliferation. When Bel-7402 cells were pretreated with OA for 24 h, OA induced apoptosis and G₂/M phase cell cycle arrest in a concentration-dependent manner. Analysis of the cell cycle regulatory proteins demonstrated that OA decreased the protein levels of cyclin B1, but increased the protein levels of p-Cdk1 (Tyr15) and p-Cdc25C (Ser 216). Moreover, OA modulated the phosphorylation of protein kinases Chk1 and p2l. Western blotting assay also showed significant decrease of Bcl-2 protein expression and increase of Bax protein expression, the cytosol Cyt c level, cleaved-caspase-9 and cleaved-caspase-3 activity. These data suggest that OA produces anti-tumor effect via induction of G₂/M cell cycle arrest and apoptosis.
Apoptosis
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drug effects
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Carcinoma, Hepatocellular
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drug therapy
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pathology
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Cell Line, Tumor
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G2 Phase Cell Cycle Checkpoints
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drug effects
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Humans
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Liver Neoplasms
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drug therapy
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pathology
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M Phase Cell Cycle Checkpoints
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drug effects
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Oleanolic Acid
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pharmacology
4.Stability analysis of allelopathic effects of Panax notoginseng on main crops by AMMI model.
Zi-long ZHANG ; Jun-ling HOU ; Wen-quan WANG
China Journal of Chinese Materia Medica 2015;40(2):191-197
This paper is aimed to study the differences of allelopathic effects of Panax notoginseng under different allelopathic chemicals resources and selection of appropriate rotation crops. The additive main effects and multiplicative interaction ( AMMI) model had been used to evaluate the stability of allelopathic effects of P. notoginseng on the varieties of corn, wheat and rice properly. The model could use not only to evaluate the stability of non-regional trial data but also explore the interaction between the rotation crop genotypes and donor substances more efficiently. Meanwhile, correspondence analysis can be used in the AMMI to evaluate genotype stability and donor substances. Ejingza No. 1 (g6) had stronger allelopathic effects with high stability, but Yunrui No. 1 (g9) which was appropriate rotation crop genotype, had weaker allelopathic effects with high stability. These findings will aid in choosing appropriate rotation crops and establishing proper rotation system.
Allelopathy
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Crops, Agricultural
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Panax notoginseng
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chemistry
5.Study on pharmacokinetic of vancomycin in normal and endophthalmitis eyes in rabbit
Ya-ling, WANG ; Hai-tao, YU ; Xiao-long, CHEN
Chinese Journal of Experimental Ophthalmology 2011;29(12):1111-1115
Background Vancomycin has been increasingly recommended for the management of endophthalmitis,but few research report has been published about the pharmacokinetics of intravitreal vancomycin up to now.It is necessary to have an exact method to measure the concentration of vancomyein in animal eyes after intravitreal injection.Objective This study was to observe and compare the phamacokinetical process of vancomycin in serum,vitreous and aqueous humor between normal and infected rabbit eyes.Methods Seventy-two healthy adult rabbits were randomly divided into normal group and infected group and 36 rabbits for each.The animal models of endophthalmitis were established by intravitreal inoculation of 2000 CFU/ml staphylococcus aureus in the right eyes of rabbits in the infected group.Once endophthalmitis developed,0.1 ml vancomycin ( 10 g/L) solution was injected into the vitreous of every rabbit.The peripheral blood,vitreous and aqueous humor samples were respectively collected in 4 rabbits for each group at 0.5,2,4,6,12,24,48,72 and 84 hours after injection for detection of vancomycin concentration by high performance liquid chromatography(HPLC-UV).3p97 software was used to create fit parameters of pharmacokinetics.This experiment followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission (Version 1988).Results The accuracy of HPLC fitted the detecting request of biological specimen.The concentration-time data of vancomycin in normal rabbit aqueous humor and vitreous was subject to two-compartment model.The pharmacokinetic parameters were separately as following:Cmax was 50.16 mg/L and 751.42 mg/L,t1/2was 51.04 hours and 53.21 hours.The concentration-time data of vancomycin in infected rabbit aqueous humor and vitreous was subject to one-compartment model.The pharmacokinetic parameters were separately as following:Cmaxwas 24.94 mg/L and 687.66 mg/L,t1/2was 11.42 hours and 12.91 hours.The concentration of vancomycin in serum was much lower and almost undectable.The concentration of vancomycin in vitreous was gradually reduced as the prolong of time after injection in both normal group and infected group,but a obvious decline after increased level was scen in aqueous humor.Compared with normal group,the concentrations of vancomycin in both vitreous and aqueous humor were reduced at various time points(P<0.05,P<0.01 ).Conclusions HPLC is simple,highly sensitive and specific for the pharmacokinetic analysis of vancomycon.These results indicate that pharmacokinetic parameters of vancomycin alter in pathological condition,which is helpful for us to establish the better treatment guidelines for endophthalmitis.
6.Preventive and Therapeutic Effects of Shenlixin Granules on Chronic Renal Failure Rats Caused by Adenine
Tongsheng WANG ; Zijiang LONG ; Jinling HUANG ; Liang WANG ; Weisheng WANG ; Ling ZHANG
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(06):-
Objective To investigate the preventive and therapeutic effects of Shenlixin Granules on chronic renal failure rats.Methods The chronic renal failure rat models were caused by adenine.After the treatment of Shenlixin Granules,the body weight,urine volume and renal index of models were examined.Results Shenlixin Granules could obviously inhibit the decrease of body weight and urine volume,reduce the renal index and had obvious protective effect on the renal tissue.Conclusions Shenlixin Granules has a preventive and therapeutic effect on chronic renal failure.
7.Total laparoscopy to treat hepatolithiasis using gallbladder-hepatic duct subcutaneous tunnel
Tao LI ; Long CUI ; Gang WANG ; Xiaofeng LING ; Chunsheng HOU ; Lixin WANG ; Zhi XU
Journal of Peking University(Health Sciences) 2016;48(5):915-918
SUMMARY To investigate the effect and feasibility of total laparoscopy to treat hepatolithiasis using gall-bladder-hepatic duct subcutaneous tunnel.Retrospective analysis was conducted of the case data of 11 pa-tients with hepatolithiasis who underwent total laparoscopic treatment using gallbladder-hepatic duct sub-cutaneous tunnel from January 2010 to October 2014.The operation time,blood loss,postoperative com-plications and recurrence of stones were recorded.All the cases completed the operation.The average hos-pital-stay was 9.2 days (range:3 -29 d).The average operation time was 298 min (range:225 -480 min).The average blood loss was 253 mL (range:50 -700 mL),and the average blood loss of liver re-section groups was 325 mL (range:200 -700 mL).The average discharge time was 3.3 days (range:3 -5 d).The rate of postoperative residual stones was 36.4% (4 /11).We extracted stones with chole-dochofiberscope via T-tube sinus six weeks after operation.One case developed biliary leakage,and healed through adequate drainage and the T-tube was pulled out after one month.There was no periopera-tive mortality.All the cases were followed up and the mean follow-up was 22 months (range:2 -51 months).The anastomotic stenosis of gallbladder-hepatic duct was found in one case.But we got a good therapeutic result with performed gallbladder chemical ablation with 95% ethanol.No recurrence of hepa-tolithiasis was found.As a choice for minimally invasive method to hepatolithiasis using gallbladder-he-patic duct subcutaneous tunnel,total laparoscopy is a safe and feasible procedure.
8.Effects of stromal cell-derived factor-1 on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cells
Quan WEN ; Yuming ZHAO ; Yuanyuan WANG ; Xu WANG ; Long LING ; Lihong GE
Journal of Peking University(Health Sciences) 2016;48(1):23-29
Objective:To compare the effects of stromal cell-derived factor-1 (SDF-1 )and granulocyte colony-stimulating factor (G-CSF)on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cell (DPSC)in vitro.Methods:DPSCs were cultured in vitro and treated with either 1 00 μg/L SDF-1 or 1 00 μg/L G-CSF.Cell counting kit-8 (CCK-8 )and colony-forming unit (CFU ) were used to detect the effect of SDF-1 and G-CSF on the proliferation ability of DPSC.Cell migration of DPSC was determined by wound healing assay and Transwell migration assay.The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP)staining, ALP activity and alizarin red S staining.The expression of odontoblastic-related genes such as dentin ma-trix protein 1 (DMP-1 )and dentin sialophosphoprotein (DSPP)were quantified by real-time RT-PCR. Results:SDF-1 and G-CSF promoted the proliferation of DPSC slightly,but the difference was not statis-tically significant.Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01 ),but there was no significant difference between the two factors.In Transwell migration assay,the number of migrated cells of the control group was 5 .0 ±1 .4 per sight,while the SDF-1 group was 24.3 ±6.8 per sight and the G-CSF group was 1 1 .8 ±3.3 per sight,suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF,and SDF-1 was more effective than G-CSF (P<0.05 ).Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining.Higher ALP activity,more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment.Conclusion:SDF-1 had no significant effect on the proliferation of DPSC,but could significantly promote cell migration and odontoblastic differentiation of DPSC.Its effect on DPSC was bet-ter than G-CSF.
9.Expression, purification and effect of different immunization approaches of Tir from EHEC O157:H7
Ling WANG ; Liping HU ; Beiguo LONG ; Yong ZHOU ; Jun LUO ; Guangming YUANG ; Hongying FAN
Chinese Journal of Microbiology and Immunology 2011;31(4):335-340
Objective To clone and express translocation intimin receptor(Tir)of enterohemorrhagic Escherichia coli(EHEC)O157:H7,and to analyze the effect of different routes on the induction of immunity to the recombinant protein.Methods The tir eucoding genes were amplified from EHEC O157:H7 strain guangzhou 246 genome,and genes were cloned into the vector pET-30a(+).The pET-30a(+)tir recombinant was transformed into E.coli BL21.and expression was induced bv IPTG.The expressed product was analyzed by SDS-PAGE and purified by Ni-IDA affinity chromatography.The immunized mice sera and fecal against the recombinant protein was detected.Resuits The length of the tir is 1677 bp,with the initiation codon ATG and the termination codon TAA.Double enzyme digestion and DNA sequencing confirmed that the recombinant expression plasmid pET-30a(+)-tir was constructed.The recombinant protein was expressed in Escherichia coli expression system,and was purified by Ni-IDA affinity chromatography.The mice were able to produce a high serum IgG antibody titer after both subeutaneous and intranasal immunizations.Meanwhile,the intranasal immunization induced serum and fecal IgA antibody titer was significantly higher than that of the subcutaneous immunization group.Conclusion Tir molecule is potential vaccine candidate for preventing EHEC disease.
10.In vitro female germline potential of human umbilical cord-derived matrix stem cells
Caixia LI ; Fengying WANG ; Zhiqing LIANG ; Yuyan LI ; Chiyang YU ; Qing CHANG ; Ling LONG
Chinese Journal of Tissue Engineering Research 2010;14(40):7583-7587
BACKGROUND:Bone marrow mesenchymal stem cells(BM-MSCs)have been shown to possess the potential to differentiate into oocytes.However,immune rejection and a limited number of donors of BM-MSCs constrain the applications of BM-MSCs.Several studies have demonstrated that human umbilical cord matrix stem cells(UC-MSCs)also have an intrinsic ability to differentiate into oocyte-like cells in vitro.OBJECTIVE:To establish the method for UC-MSCs culture and to investigate the in vitro differentiation potential of UC-MSCs towards germ cells.METHODS:Umbilical cord from full-term normal deliveries was obtained in sterile condition.Collagenase I-digested cells were cultured in DMEM.The immunophenotype of cells was determined by flow cytometry.Lipoblasts,osteoblasts and chondroblasts were induced in different condition cultures.The expression of germ cells specific marker in UC-MSCs was determined by reverse transcdption-polymerase chain reaction.Follicular fluid was employed to induce UC-MSCs differentiation into germ cells.RESULTS AND CONCLUSION:Spindle-like umbilical cord cells were shown and cells in culture were extended to more than 10passages.BM-MSCs-like immunophenotypes were shown:CD29,CD44,CD73(SH3),CD90 and CD105(SH2)were positive;SSEA-4 was weakly positive;CD31,CD34,CD45 and HLA-DR were negative.After UC-MSCs were induced in different condition cultures,lipid droplet-,bone tubercle-,and cartilage tubercle-like structures emerged and the mRNA expressions of specific gene of fat,bone and cartilage were observed.Germ cells markers,OCT4,Stella,Ifitm3,were expressed in UC-MSCs.After induced by 5%,10% or 20% follicular fluid,cells aggregated and oocyte-like structures were observed.Human UC-MSCs could be cultured and amplificated in vitro.UC-MSCs showed immunophenotypes similar to BM-MSCs.UC-MSCs had the potential to differentiate into lipoblasts,osteoblasts,and chondroblasts.Oocyte-like structure was induced in vitro from UC-MSCs with germ cells specific marker.These findings suggest that UC-NSCs have the potential to differentiate into germ cells.