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The aim of this work is to discover the inhibitory mechanism of tea peptides and to analyse the affinities between the peptides and the angiotensin-converting enzyme (ACE) as well as the stability of the complexes using in vitro and in silico methods. Four peptide sequences identified from tea, namely peptides I, II, III, and IV, were used to examine ACE inhibition and kinetics. The half maximal inhibitory concentration (IC
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BACKGROUND:Nickel-titanium memory-shape compression anastomosis clip (Ni-Ti CAC) has been used in gastrointestinal anastomosis, but its efficacy and safety still remain controversial.OBJECTIVE:To evaluate the efficacy and safety of Ni-Ti CAC in gastrointestinal anastomosis.METHODS:A computer-based online research of PubMed, EMbase, Cochrane Library, CBM, CNKI, and VIP databases was performed for articles published before January 15th, 2017 using the keywords of nickel-titanium, compression anastomosis clip, anastomosis, gastric, jejunum, ileum, small intestine, colon, rectum, and large intestine in English and Chinese, respectively. The randomized controled trials about Ni-Ti CACversus conventional methods for gastrointestinal anastomosis were included. Meta-analysis of the anastomosis time, time of exsufflation, and hospitalization time was performed, and sequential analysis was conducted on TSA v0.9 software.RESULTS AND CONCLUSION:A total of 18 eligible randomized controlled trials were enroled, involving 1860 patients. Ni-Ti CAC could reduce the anastomosis time [MD=-3.83, 95%CI(-6.48,-1.19),P=0.004] and time of exsufflation [MD=-0.14, 95%CI(-0.22,-0.05),P=0.002], but there was no significant difference in the hospitalization time [MD=-0.83, 95%CI(-1.82, 0.16), P=0.10]. The quality was ranked as low level based on GRADE system. The time of exsufflation of Ni-Ti CAC was superior to that of conventional method, which was confirmed by sequential analysis. One case of death was reported and incision infection was the most common adverse effects; additionaly, pulmonary embolism and abdominal pain occurred. To conclude, Ni-Ti CAC can facilitate gastrointestinal anastomosis, accelerate the time of exsufflation, and holds a good safety. However, more multicenter and high-quality randomized controlled trials are needed.
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Objective:To study the therapeutic effect of a novel double-target system,in which human umbilical cord-derived MSCs were used as vehicles to deliver fusion protein scFvCD20:sTRAIL to non-Hodgkin ’ s lymphoma. Methods: The traditional methods in molecular biology were used to construct lentivirus expression vectors pLenR. scFvCD20: sTRAIL and contrast vectors. Human umbilical cord-derived MSCs ( HUMSCs ) were labeled with the copGFP by transducing with pseudo viral particles which had been packaged in 293T cells with four plasmid-lentivirus packaging system. Fusion protein scFvCD20:sTRAIL were secreted from MSC. scFvCD20:sTRAIL after that HUMSCs were infected by pseudo viral particles. CCK8 assay was applied to detect the antigen-restricted cell death induced by scFvCD20:sTRAIL in CD20-positive BJAB and Raji cells as well as CD20-negtive Jurkat cells and human normal peripheral blood mononuclear cells (PBMCs). To evaluate the therapeutic effect of MSC. scFvCD20:sTRAIL in vivo,ge-netically modified HUMSCs were intravenously injected into tumor-bearing mice with BJAB cells. The volume of tumor was measured every three days, and the inhibition ratio of tumor was calculated according to tumor volume. Results: Lentivirus expression vectors pLenR. scFvCD20:sTRAIL, pLenR. ISZ:sTRAIL, pLenR. scFvCD20 and pLenR. CopGFP were successfully constructed and these constructs could be expressed stably in HUMSCs by lentivirus transduction. scFvCD20:sTRAIL fusion protein produced a potent inhibition of cell proliferation in CD20-positive BJAB cells,moderate inhibition of the growth of Raji cells,and weak inhibition in CD20-negtive Jurkat cells when compared with ISZ-sTRAIL treatment,and it had no effect on normal human peripheral blood mononuclear cells (PBMCs). The MSC. scFvCD20:sTRAIL treatment significantly inhibited the tumor growth when compared with those treated with MSC. ISZ-sTRAIL. Conclusion: A double-target therapeutic system is well established, in which HUMSCs migrated to tumor site, secreted a novel fusion protein scFvCD20:sTRAIL,and thus locally concentrated scFvCD20:sTRAIL extended antigen-restricted anti-tumor activity. The engineered HUMSCs secreting scFvCD20:sTRAIL showed potent effect on inhibiting tumor growth in BJAB lymphoma malignancy,which may play an essential role in the clinical research .
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Objective To explore β-sodium aescinate on vascular endothelial function ( FMD ) , homocysteine ( Hcy ) and hypersensitive C-reactive protein ( hs-CRP) and clinical efficacy in patients with acute cerebral infarction.Methods 198 acute cerebral infarction patients from March 2013 to April 2015 were randomly divided into observation group (n=100) and control group (n=98).Control group were treated according to the condition of the disease, observation group were treated by β-sodium aescinate base on control group, 20mg was added to 250mL saline for intravenous drip,one times per day.Continuous used 14d for one treatment courses.Compared the change of vascular endothelial function, Hcy and hs-CRP and clinical efficacy.Results The total effective rate of observation group was 90.00%, which was significantly higher than that of 71.42% in control group (χ2 =11.01,P<0.05).Post-treatment the value of FMD significantly increased, Hcy and hs CRP were significantly decreased both in observation group and control group respectively, which the difference had a statistically significant as compared with Pre-treatment (P<0.05);but, the value of FMD was significantly higher, Hcy and hs CRP was significantly lower in observation group than that of control group (P<0.05).Conclusion It has a significant β-sodium aescinate clinical effect in treatment of acute cerebral infarction, and FMD are significantly higher, Hcy and hs-CRP are significantly decrease.
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Aim Drug resistance is one of the major hinders on cancer treatments. α-enolase ( eno1 ) was closely related to the generation and development of drug resistance. This article aims to study the effect of eno1 on cell growth and drug resistance in human chro-nic myeloid leukemia cell line K562/A02 . Methods We screened three eno1 stable silencing cells K562/A02-sheno1 and its control cells K562/A02-shcon. Cell count assay was performed to test cell growth, MTT assay was used to test cell proliferation, flow cytometry was used to test the intra-cellular Rho123 content, the expression of genes were tested by real-time PCR assay and western blot assay on mRNA level and protein level, respectively. Results eno1 was o-ver-expressed in K562/A02 cells and its expression was increased by ( 2. 85 ± 0. 56 ) times and ( 1. 43 ± 0. 05 ) times on mRNA level and protein level com-pared to K562 cells. However, there was no difference in cell growth rate between K562/A02 cells and K562 cells. K562/A02-sheno1 cells showed lower cell growth rate and higher drug sensitivity to anti-cancer drugs taxol and doxorubicin. Moreover the Rho123 content was increased in K562/A02-sheno1 cells. The expression of MDR1 decreased in both mRNA level and protein level in K562/A02-sheno1 cells. Conclusion eno1 silencing could suppress cell growth, reverse drug resistance and increase its drug sensitivity in K562/A02 cells, and the mechanism was associated with the MDR1 gene.
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Aim To investigate the expression of AB-CB5 and MDR1 in the cell line KG1 a and samples from acute myeloid leukemia ( AML) and their effects on multidrug resistance. Methods The expression of ABCB5 and P-gp ( the expressed product of MDR1 ) in KG1 a cells were detected by flow cytometry as well as Western blot analysis; KG1 a cells were transfected with the specific siRNA of ABCB5 using lipo2000 to reduce the expression of ABCB5; intracellular rhoda-mine123 was measured by flow cytometry;cell viability was detected by MTT; the expressions of ABCB5 and MDR1 in samples from AML were detected by real time PCR. Results ABCB5 and P-gp were overexpressed in KG1 a;the specific siRNA of ABCB5 transiently in-hibited the expression of ABCB5 in KG1 a; the siAB-CB5-KG1 a cells increased the intracellular rhodamine 123 and have been more sensitive to adriamycin com-pared with the parent KG1a. ABCB5 gene expression in samples from AML was higher than healthy people. Further, the expression of ABCB5 in 38 relapse or re-fractory AML significantly exceeded the 33 drug sensi-tive. And we found a significant positive correlation between ABCB5 expression and MDR1 gene expression in the 38 patients with relapse or refractory AML. Conclusion ABCB5 , as well as P-gp contributes to mediate multidrug resistance of AML, which provides a novel target for the therapy of relapse or refractory AML.
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Aim To study targeting capability of anti-CD19 (Fab)-LDMto CD19 +B lymphoma cells in vi-vo and in vitro.Methods Flow cytometry was em-ployed to determine the affinity of Cy5 labeled anti-CD19 (Fab)-LDP to human lymphoma Raji cells.And the optical imaging system was used to analyze the dis-tribution of Cy5-anti-CD19 (Fab )-LDP in lymphoma-transplanted xenograft nude mice in vivo.Results The results of flow cytometry demonstrated that Cy5-an-ti-CD19(Fab)-LDP had remarkable affinity with lym-phoma Raji cells;Raji lymphoma xenograft model was established successfully in nude mice and in vivo fluo-rescence imaging analysis indicated that the antibody-drug conjugates could specially be localized in the tar-get tumor.Conclusion The experiments in vivo and vitro confirm that anti-CD19 (Fab)-LDP has remarka-ble affinity to targeting CD19 +lymphoma cells,and the antibody drugs anti-CD19 (Fab )-LDP have the probability to be new drugs for the treatment of malig-nant lymphoma.
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Objective To explore the key points of nursing for neonates with hypoxic-ischemic encephalopathy detected by 18F-fluorodeoxyglucose positron emission tomography.Methods The features of nursing for 58 cases neonates with hypoxic-ischemic encephalopathy detected by 18F-fluorodeoxyglucose positron emission tomography were analyzed and summarized.Results 55 cases with hypoxic-ischemic encephalopathy detected by 18F-fluorodeoxyglucose positron emission tomography smoothly with pertinent nursing,only 3 cases had not completed the procedures because of change of their conditions.Conclusions Pertinent nursing was important for neonates with hypoxic-ischemic encephalopathy detected by 18F-fluorodeoxyglucose positron emission tomography because it could help them complete the procedures smoothly and make the results more accurate and satisfying so it is worthy of wide use clinically.
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Objective To construct a fusion protein that used for treatment of resistance and palindromia in leukemia and studied its biological activity. Methods IL-3 and LP gene fragments were amplified by PCR. After enzymatic digestion and T4 ligation, the fusion gene was cloned into expression vector pAYZ. The product was purified by exchange chromatography and anti-Etag affinity chromatography. IL3-G4SLP fusion protein was analyzed by SDS-PAGE and Western blot. Protein biological activity was detected by FACS. Results The fusion protein was expressed as soluble protein by E.Coli 16C9. The protein expression level was about 1 mg/L, its purity was over 95 %, and the expression level was about 1 mg/L. The fusion protein can combined specificely with CD123 on leukemia stem cells. Conclusion Fusion protein IL-3-G4S-LP can target on leukemia stem cells and maybe as a potential drug used for treatment of resistance and palindromia in leukemia.
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Aim To study the mechanism of synergistic antitumor of EBB and doxorubicin in doxorubicin-resistant MCF-7/ADR breast carcinoma cells.Methods The antitumor activity of doxorubiein alone and its combination with EBB were measured by MTT assay in MCF-7/ADR and MCF-7cells. The rate of doxorubicin-induced apoptosis and the protein and mRNA levels of P-glycoprotein(P-gp) were determined in MCF-7/ADR treated with EBB by flow cytometry (FACS), respectively.Laser scanning confocal microscopy was used to detect the intracellular accumulation of drug in EBB-treated MCF-7 and MCF-7/ADR cells.Results EBB had antitumor effects for MCF-7 and MCF-7/ADR.It could potentiate the antitumor effect of dororubicin with CDI of 0.73 and 0.49 for MCF-7 and MCF-7/ADR,respectively.EBB and doxorubicin acted synergistically in elevating apoptosis of MCF-7/ADR and downregulating the expression of P-gp in a dose-dependent manner in MCF-7/ADR.EBB restored the intracellular accumulation of doxorubicin in MCF-7/ADR cells in a dose-dependent manner.After pretreatment with EBB for 24 h and 48 h,the intracellular accumulation of doxorubicin and Rh123 was obviousely restored in MCF-7/ADR cells compared with control in a time-dependent manner.Conclusion EBB is a potential agent which has strong inhibitory effect on both multidrug resistant cells and their parental cells.EBB can significantly potentiate the antitumor effects of dororubicin in MCF-7/ADR cells by blocking the function of P-glycoprotein and inhibiting the expression of P-glycoprotein.
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Objective To investigate the change of high mobility group box 1 ( HMGBI ) after intestine ischemia reperfusion (I/R) in rats, compare the effect of drainage of intestine lymph fluid on gut barrier, and ex- plore the possible mechanism of iachemia-reporfusion injury. Methods Thirty-two Sprague-Dawley (SD) rats (SPF grade) were randomly divided into4 groups with 8 rats in each group: blank group, sham group, intestine is-chemia-reperfusion (I/R) group, and intestine ischemia-reperfusion with drainage of intestine lymph fluid (I/R +drainage) group. Indicators of gut barrier function damage, translocation of endotoxin, and change of HMGB1 and cytokines were detected after intestine ischemia-reperfusion injury. Results The gut barrier function damage and levels of endotoxin, HMGBI, tumour necrosis factor-alpha ( TNF-α), interleukin-6 ( IL-6 ), interleukin-1 beta (IL-1β), and soluble intercellular adhesion molecule-1 (sICAM-1) were significantly lower in blank group and sham group than in I/R group and I/R + drainage group ( P < 0. 05 ). Compared with the intestine I/R + drainage group, the levels of endotoxin and cytokines were significantly higher in the intestine I/R group. The level of HMGB1 was slightly higher than that in the intestine I/R + drainage group, but such difference was not statistically significant ( P > 0. 05 ). lmmunohistochemical staining also revealed that the expression of HMGB1 was significant- ly higher in I/R group than in I/R + drainage group. Conclusions Intestine iachemia-reperfusion injury can lead to the injury of intestine mucosal barrier and increase HMGB1 level HMGB1 may deteriorate gut barrier function and increase the leveh of systemic cytokines. Drainage of lymph fluid can block the gut-lymph pathway and thus decrease the levels of endotoxin and cytokines in systemic circulation and attenuate intestine ischemia-reperfusion injury.
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Aim To construct and express anti-CD20Fab-LDP,generate anti-CD20Fab-LDM and identify its biological activity.Methods PCR and overlapping PCR were used to construct anti-CD20Fab-LDP.DNA sequence was analyzed by the Terminus of Dideoxy Nucleotide.The product was purified by affinity chromatography and analyzed by Western blot and its antigen-binding activity was examined by FACS.Specific killing activity in vitro of anti-CD20Fab-LDM was analyzed by MTT.Results The data of DNA sequence showed that anti-CD20Fab-LDP was correct.The fusion protein was recovered in high yield(up to 4 mg·L~(-1))after proteinG purification.The fusion protein could bind to Raji cells(CD20+),and similar affinity data were obtained with anti-CD20Fab.Anti-CD20Fab-LDP showed potent cytotoxicity to Raji cells with IC_(50) values of 0.9×10~(-10) mol·L~(-1).Conclusions Anti-CD20Fab-LDP with high level expression was successfully obtained and could bind to Raji cells cells.Anti-CD20Fab-LDM showed specific killing activity to Raji cells in vitro.
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Background and purpose:Bispecific antibody is a special pattern of genetic engineering antibody.It possesses 2 antigen binding sites, one directed at the effecter cells and the other at the target cells which are always the tumor cells.This enables the effecter cells to recruit to the targets efficiently and exert cytotoxic effect.The experiment was aimed to construct the expression vector of anti-CD3?anti-CD19 bispecific antibody, purification and analyzing the activity of the diabody.Methods:Applyed PCR and overlap PCR to construct the expression vector of anti-CD3?anti-CD19 bispecific antibodies, transformed it into E.coli 16C9 for expression.Purified the diabody by His-tag affinity chromatography and confirmed by 12% SDS-PAGE and Western blot.Antigen binding activity of the diabody was analyzed by FACS.Results:The DNA sequence of the expression vector containing anti-CD3?anti-CD19 fragment was comfirmed.The yield of purified diabody was up to 5 mg/L.The molecular weight of diabody was correctly indicated by SDS-PAGE and Western blot.The diabody could specifically bind to CD19+ Raji cells and CD3+ Jurkat cells while competitively inhibit binding of HIT3a and HIT19a to the cells.Conclusion:Successfully constructed the expression vector of anti-CD3?anti-CD19 bispecific antibodies, the diabody acquired by the method could be produced with high level expression and also had high specific affinity with CD19+ Raji cells and CD3+ Jurkat cells.