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Objective:To identify the factors associated with long-term survival and guide the decision for primary surgery in patients with advanced high-grade serous ovarian cancer(HGSOC).Methods:In this case-control study, clinical parameters, including surgical and non-surgical associated factors, were collected and compared between the patients with short-term (<2 years) and long-term (>5 years) survival who all underwent primary debulking surgery (PDS) followed by carboplatin and paclitaxel chemotherapy from January 2004 to December 2016. Univariate analysis was examined by chi-square test and multivariate analysis was performed by logistic regression analysis.Results:There were 95 cases long-term survival (LTS group) and 77 cases short-term survival (STS group) in 698 newly diagnosed HGSOC patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ⅲc and Ⅳ who met include and exclude criteria. (1) Univariate analysis showed that the proportion of complete cytoreduction with no visible residual disease (R0) at PDS and platinum sensitivity in LTS group were significantly higher than those in STS group ( P<0.01). The surgical complexity score (SCS), the preoperative serum CA 125 level and the ascites volume in the LTS group were significantly lower than those of the STS group (all P<0.05). In the LTS group, the preoperative incidence of lesions in retrograde peritoneum of the bladder, serosal and mesangial membrane of the small intestine, upper abdominal peritoneum and liver parenchyma were significantly lower than those in the STS group (all P<0.05). Multivariate logistic regression analysis showed that platinum sensitivity ( OR=0.016, 95% CI: 0.004-0.063, P<0.01), ascites volume >500 ml ( OR=3.193, 95% CI: 1.285-7.930, P=0.012), and SCS ≥8 ( OR=17.433, 95% CI: 2.281-133.25, P=0.003) were independent factors affecting long-term survival ( P>0.05). (2) Totally 37 of 95 in long-term survival and 16 of 77 in short-term survival achieved R0 cytoreduction at PDS. Univariate analysis showed that preoperative serum CA 125 level, preoperative lesion score, preoperative lesion (DS) score, ascites volume, platinum sensitivity,and SCS were significantly correlated with the R0 PDS (all P<0.05). Multivariate analysis showed that ascites volume >500 ml ( OR=5.199, 95% CI: 2.015-13.409, P=0.001), DS >2 ( OR=15.264, 95% CI: 5.843-39.874, P<0.01) and SCS ≥4 ( OR=4.176, 95% CI: 1.618-10.777, P=0.003) were independent factors associated with R0 cytoreduction. In patients with DS ≤2 or SCS <4, but not those with DS >2 or SCS ≥4, R0 cytoreduction was significantly associated with long-term survival. Conclusion:The intrinsic biology of tumor is the factor influencing long-term survival of advanced HGSOC patients, and those who present with wide intraperitoneal metastases and need to remove multiple organs may not benefit from R0 cytoreduction.
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<p><b>OBJECTIVE</b>To obtain streptavidin-tagged human granulocyte-macrophage colony-stimulating factor (SA/hGM-CSF) fusion protein and evaluate its bioactivity .</p><p><b>METHODS</b>PET24a-6His-SA-L-hGM-CSF and PET24a-hGM-CSF-L-SA-6His plasmids were constructed and expressed in Rosetta (DE3) host bacteria to generate the fusion proteins. The two fusion proteins were refolded by gradient dialysis after Ni-NTA affinity chromatography and finally purified using DEAE-sepharose FF anion exchange chromatography. MTT method was used to evaluate the effect of SA/hGM-CSF fusion proteins in inducing the proliferation of human erythroleukemia cells (TF-1). The efficiency of the fusion proteins for surface modification of biotinylated MB49 tumor cells was evaluated by flow cytometry.</p><p><b>RESULTS</b>The recombinant fusion proteins SA-hGM-CSF and hGM-CSF-SA were highly expressed in Rosetta (DE3) at about 20% of the total bacterial proteins, with a purity of about 96% after purification. The two fusion proteins exhibited bifunctional activities, namely the pro-proliferation effect on human erythroleukemia cells (TF-1) and SA-mediated high-affinity binding to biotinylated cell surfaces (with an anchoring modified rate of about 99%).</p><p><b>CONCLUSION</b>SA/hGM-CSF bi-fusion proteins obtained in this study lays the groundwork for the development of cancer cell vaccines with surface modification by hGM-CSF.</p>
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Humans , Biomarkers , Cancer Vaccines , Cell Line, Tumor , Diterpenes , Pharmacology , Escherichia coli , Metabolism , Granulocyte-Macrophage Colony-Stimulating Factor , Membrane Fusion Proteins , Plasmids , Streptavidin , ChemistryABSTRACT
AIM: To investigative the inhibitory effects of Paecilomyces cicadae polysaccharide (PcPS) against HBV in vitro, and the effects on the Toll like receptor 4 (TLR4) mRNA expression in HepG2.2.15 cell strain. METHODS: HepG2.2.15 cell strain was co-cultured in vitro with PcPS in different concentrations, and lamivudine (LMV) was applied as positive control. MTT assay was employed to detect the cytotoxicities of PcPS in vitro, when the HepG2.2.15 cells was used as target cells. The effects of PcPS on the secretion of HBsAg and HBeAg were assayed by ELISA method. Fluorescence quantitative-PCR (FQ-PCR) was used to detect the inhibitory effects of PcPS on the content of HBV-DNA and TLR4 mRNA in HepG2.2.15 cells. RESULTS: The inhibitory effects of PcPS on the HBsAg and HBeAg were observed and the maximum inhibitory ratio up to 44.8% and 31.0%, respectively. The same inhibitory effects of PcPS on the HBV-DNA replication and TLR4 mRNA expression in HepG2.2.15 cells were also found. CONCLUSION: A certain concentration of PcPS significantly inhibits HBV replication in vitro.
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AIM: To investigate the effect of puerarin on diabetic myocardial damage and to explore its possible mechanisms in rats. METHODS: The male Sprague-Dawley rats were randomly divided into normal control group, diabetic group, high dose puerarin (160 mg/kg), middle dose puerarin (120 mg/kg), low dose puerarin (80 mg/kg) treatment groups and aminoguanidine (100 mg/kg) treatment group. Corresponding drugs were intraperitoneally injected once a day. The animals in normal control group and diabetic group were given equal propylene glycol. 12 weeks later, the rats were sacrificed and their cardiac muscles were collected. Myocardial structure was observed under transmission electron microscope (TEM). The blood glucose concentration and the activities of superoxide dismutase (SOD), Ca~(2+)-ATPase, Na~+-K~+-ATPase and the content of malondialdehyde (MDA) in myocardial homogenate were measured biochemically. In addition, the mRNA expressions of peroxisome proliferator activated receptor γ (PPAR-γ), glucose transporter-4 (GLUT-4) and aldose reductase (AR) in myocardial tissues were determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: It is observed that the myofibrils were diminished, broken or fused, some lipid droplets deposited in the cytoplasm of cardiocytes and part of cristae of mitochondria were broken or disappeared under TEM in diabetic group. The activities of SOD, Ca~(2+)-ATPase, Na~+-K~+-ATPase as well as the mRNA expressions of PPAR-γ and GLUT-4 decreased significantly (all P<0.01). The blood glucose concentration and the MDA level and the AR mRNA expression increased obviously (all P<0.01) in diabetic group as compared to those in normal control group. How-ever, in puerarin treatment groups, the above changes were reversed, a significant differences of those were found as compared to those in diabetic group (P<0.05 or P<0.01, respectively). The pathological change of cardiac muscles was relieved. It showed that myofibrils were well-arranged and only few lipid droplets deposited in the cytoplasm of cardiocytes and most mitochondria had clear and regular cristae under TEM in puerarin treatment groups. CONCLUSION: Puerarin exerts preventive and remedial effects on the diabetic myocardium, which may be related to up-regulating the mRNA expressions of PPAR-γ and GLUT-4, promoting glucose uptake and relieving oxidative stress damage.
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Based on the practice of bilingual education in medical biochemistry,this paper discussed the advantages,problems and solutions of bilingual education in medical biochemistry of teaching-centred medical college or university.This study will be helpful for developing bilingual teaching of biochemistry in other teaching-centred medical colleges or universities.
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AIM:To investigate the protective effect of puerarin on testis in diabetic rats.METHODS:The male Sprague-Dawley rats were randomly divided into normal control group,diabetic group,high dose puerarin(160 mg/kg),middle dose puerarin(120 mg/kg),low dose puerarin(80 mg/kg)treatment groups and aminoguanidine(100 mg/kg)treatment group.Corresponding drugs were intraperitoneal injected once a day.The animals in normal control group and diabetic group were given equal propylene glycol.12 weeks later,the rats were sacrificed and their testes were collected.Testicular structure was observed under transmission electronic microscope(TEM).The contents of nitric oxide(NO),malondialdehyde(MDA)and the activity of nitric oxide synthase(NOS),inducible nitric oxide synthase(iNOS),superoxide dismutase(SOD),Ca2+-ATPase and Na+-K+-ATPase in testis homogenate were detected biochemically.The levels of endothelin-1(ET-1)and calcitonin gene-related peptide(CGRP)were also measured by radioimmunoassay.In addition,the mRNA expressions of eNOS,iNOS,CGRP,ET-1 and AR in testis tissues were determined by reverse transcription polymerase chain reaction(RT-PCR).RESULTS:The pathologic changes such as cytoplasmic inclusion of sertoli cell and obvious vacuoles between sertoli cells and basal membrane of the contorted seminiferous tubules were observed under TEM in diabetic group.The contents of NO and CGRP,the activities of SOD,Ca2+-ATPase,Na+-K+-ATPase as well as the mRNA expressions of eNOS and CGRP decreased significantly(P
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AIM: To study the immune regulation of paecilomyces cicadidae total polysaccharides. METHODS: After subcutaneous injection with 200 mg/L paecilomyces cicadidae total polysaccharides in the back of the rats for 17 days, the white blood cell (WBC) counts, and the levels of lipid peroxide (LPO) and reduced glutathione (GSH) in the spleen and thymus of rats were detected. The alveolar macrophages (AM?) cultured with 10% fetal bovine serum-RPMI-1640 for 2 h in vitro , then the activity of lactate dehydrogenase (LDH) and acid phosphatase (ACP) were detected by automatic biochemistry analyzer, and the ability devouring neutral red in the AM? were examined. RESULTS: Compared with control group, the WBC, the levels of GSH in the spleen and thymus, and the activities of LDH and ACP were significantly increased and the ability of devouring neutral red was also strengthened( P
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AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTM). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTM were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-?) and interleukin-6(IL-6) in the hTM was analyzed by radioimmunoassay, after hTM were cultured with 0.312,0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTM induced by PPM, and the levels of TNF-? and IL-6 in the hTM induced by PPM increased markedly too. And the induced expression effect of TNF-? and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTM,and activate hTM in vitro.
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AIM: To investigate the protective effect of crude Paecilomyces tenuipes polysaccharides(cPtPs) and Paecilomyces tenuipes polysaccharides(PtPs) in a rat model of acute hepatic necrosis induced by carbon tetrachloride(CCl4).METHODS: Wistar rats were divided into four groups(control group,CCl4 group,cPtPs+CCl4 group and PtPs +CCl4 group),the four groups were given intragastrically with normal saline,cPtPs and PtPs for 15 d,respectively.In the last two days,these groups except control group were injected peritoneally with CCl4.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),direct bilirubin(DBIL) and indirect bilirubin(IBIL) were detected by automatic biochemistry analyzer.Pathological changes of hepatic tissue were assessed by hematoxylin-eosine(HE) staining.The levels of superoxide dismutase(SOD) and malondialdehyde(MDA) in liver homogenate were analyzed using xanthinoxidase and thio-barbituric acid,respectively.The concentration of Ca2+ in hepatocyte mitochondria was determined by colorimetric method.The expression of ?-smooth muscle actin(?-SMA) was examined in hepatic tissue by immunohistochemical method.RESULTS: Compared with control group,serum levels of ALT,AST,TBIL,DBIL and IBIL in CCl4 group increased significantly,denaturation and necrosis implicated to the whole hepatic lobules.Compared with CCl4 group,serum levels of ALT,AST,TBIL,DBIL and IBIL in PtPs +CCl4 group decreased significantly,denaturation and necrosis located in the third region of hepatic lobules,the level of SOD increased and MDA decreased(P
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Objective To investigate the physicochemical properties,structural characters and antitumor activity of a polysaccharide PTPS-1 from Paecilomyces tenuipes.Methods PTPS-1 was obtained by DEAE-cellulose and Sephadex G-100 chromatography column.The methods of HPLC,GC,IR,and NMR were used to investigate the molecular weight,monosaccharide composition,and structural characters of PTPS-1.The CCK-8 kit was used to determine the proliferation of PTPS-1 on murine splenocyte and the supernatant of the murine macrophage with PTPS-1-stimulated on Sp2/0 cells.Results PTPS-1,whose molecular weight was 1.84?104 and purity was 98.977%,contained mannose(Man) and galactogen(Gal) as monosaccharide constitute in molar ratio of 1.3∶1.It consisted of a backbone of ?-(1→6),?-(1→2),and ?-(1→3) linkage,whose branches were made by Gal-(1→3) linkage.PTPS-1 significantly stimulated the murine splenocyte proliferation and the supernatant of macrophage inhibited the growth of Sp2/0 cells in vitro.Conclusion PTPS-1 is a polysaccharide from P.tenuipes with immunomodulatory and antitumor activity.
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AIM: To study the effects of paecilomyces cicadidae total polysaccharides on the immune function and free radical scavenging of tissues in immunosuppressed rats.METHODS: The activities of lactate dehydrogenase(LDH) and arginase(Arg) in liver,kidney,spleen,thymus of immunosuppressed rats were detected by automatic biochemistry analyzer.The content of lipid peroxide(LPO) and the reduced glutathione(GSH) level in liver,kidney,spleen,thymus of immunosuppressed rats were observed.RESULTS: The activity of the LDH,Arg and GSH level were significantly increased,and the LPO content were significantly declined in liver,kidney,spleen,thymus of immunosuppressed rats induced by cyclophosphamide.CONCLUSION: Paecilomyces cicadidae total polysaccharides promote immune function and the ability of free radical scavenging of tissues in immunosuppressed rats.
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AIM: To explore the activative effect of water extract from cultured mycelium of Paecilomyces cicadae (P. cicadae) on peritoneal macrophages (PM?) and alveolar macrophages (AM?) of rats.METHODS: The rats were randomly divided into four groups: normal control group(Ⅰ), cyclophosphamide (Cy) group (Ⅱ), P. cicadae group(Ⅲ), Cy+ P.cicadae group(Ⅳ).The rats were bred in the same circumstance and were administered with corresponding drugs by subcutaneous injection for 26 days. PM? and AM? of rats were irrigated by normal saline and collected by centrifuge and incubated in a humidified incubator for 2 h at 37 ℃. The activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH) in the PM? and AM? were determined by biochemical methods, and the capability of PM? and AM? in phagocytosis of neutral red were measured by colorimetric method too. RESULTS: After administering P. cicadae to rats, the activities of ACP and LDH in PM? and AM? of normal rats were elevated significantly,and the reduction of the activities of ACP and LDH in PM? and AM? of rats due to Cy was notably antagonized, and the capability of PM? and AM? phagocytizing the neutral red were strengthened significantly.CONCLUSION: P. cicadae can activate the PM? and AM? of rats.
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Aim To evaluate the effect and mechanism of vascular endothelial growth factor(VEGF) and Cysteine protease aspartic acid-3(Caspase-3) inhibition of human hepatocellular carcinoma(HCC)implants in nude mice by nanometer magnetic interferon-?2b liposome targeting therapy.Methods Thirty nude mices bearing human HCC were divided into 5 groups of 6 mice:group 1,saline(NS);group 2,free interferon-?2b group(IFN);group 3,interfe-ron-?2b liposome group(IL);group 4,magnetic liposome group(ML);group 5,nanometer magnetic interferon-?2b liposome group(MIL).Above dosage of 200 ?l/nude mice was respectively injected through the caudal vein in these 6 groups.Meantime,magnetic field with the strength of 5000 GS was added to the tumor surface about 30 min,and this therapy was done three times.After 30 days,the mices were killed and the tumors were taken out.The difference at the mRNA expression level of VEGF and Caspase-3 was observed by RT-PCR between the experimental group and control group.The protein expression level of VEGF and Caspase-3 was detected by western blot between the experimental group and control group.Results Targeting therapy study in nude mice bearing human HCC displayed that the growth speed of tumor in the MIL group significantly slowed down than other groups.The tumor inhibition rate of MIL group was 62.50%,which was remarkably higher than those of the IFN group(27.61%) and IL group(28.17%).RT-PCR showed that the mRNA expression of VEGF of MIL group is the lowest of all groups(P
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AIM: To study the activation and the immune regulation of paecilomyces cicadidae total polysaccharides on macrophages in old rats. METHODS: The ability of devouring neutral red and stimulating index in the alveolar macrophages (AM) and peritoneal macrophages (PM) were observed. The activities of lactate dehydrogenase (LDH), acid phosphatase (ACP) and arginase (Arg) were detected by automatic biochemistry analyzer. The shape and structure of spleen macrophages were observed by electron microscope. RESULTS: Compared with control group, the activity of the LDH/ACP/Arg in the AM or PM and in the culture fluid of the AM or PM were significantly increased (P
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AIM: To explore the immunomodulatory effect of paecilomyces cicadidae polysaccharides (PCPS).METHODS: Subcutaneous injection with 50, l00, 200 mg/kg of PCPS were given in the back of the rats everyday for l5 days. The number of white blood cells (WBC) was counted. The activities of acid phosphatase (ACP) and lactase dehydrogenase (LDH)in liver, kidney, spleen and thymus were detected by automatic biochemistry analyzer. The ability of devouring neutral red and activity of ACP, LDH, arginase in alveolar macrophages were also detected. The body weight of the rat everyday during experiment and weight of the spleen and thymus after the rats were killed were measured and wet weight index was calculated.RESULTS: The wet weight index of spleen and thymus, the activity of ACP, LDH, arginase and ability of devouring neutral red in alveolar macrophages in the test group treated with PCPS were significantly higher than those in control group. CONCLUSION: PCPS shows a significant immunomodulatory effect with the increasing counts of WBC and activation of alveolar macrophages in a dose-dependent manner.