The inhibitory mechanism of copper（Ⅱ） on the aggegation of amyloid β-peptide （Aβ） was investigated by molecular dynamics simulations. The binding mode ofcopper（Ⅱ） with Aβ is characterized by the imidazole n...The inhibitory mechanism of copper（Ⅱ） on the aggegation of amyloid β-peptide （Aβ） was investigated by molecular dynamics simulations. The binding mode ofcopper（Ⅱ） with Aβ is characterized by the imidazole nitrogen atom, Nπ, of the histidine residue H 13, acting as the anchoring site, and the backbone＇s deprotoned amide nitogen atoms as the main binding sites. Drove by the coordination bonds and their induced hydrogen bond net, the conformations of Aβ converted from β-sheet non-β-sheet conformations, which destabilized the aggregation of Aβ into fibrils.展开更多
为研究TGF-β通路对猪颗粒细胞增殖、凋亡的影响,本研究对TGF-β通路基因进行调控处理,设计并筛选可高效干扰TGFβRⅠ及TGFβRⅡ表达的载体,转染不同干扰载体及添加10 ng/m L TGFβ1和25 ng/m L FSH,采用Westernblot、MTT和流式细胞法...为研究TGF-β通路对猪颗粒细胞增殖、凋亡的影响,本研究对TGF-β通路基因进行调控处理,设计并筛选可高效干扰TGFβRⅠ及TGFβRⅡ表达的载体,转染不同干扰载体及添加10 ng/m L TGFβ1和25 ng/m L FSH,采用Westernblot、MTT和流式细胞法检测不同处理对颗粒细胞TGFβ RI和TGFβ RII蛋白表达量和细胞增殖凋亡的影响。结果表明:成功筛选出干扰效率最高的p OSP1-TGFβRⅠ-sh5和p OSP1-TGFβRⅡ-sh3载体,对TGFβRⅠ和TGFβRⅡ的抑制率分别为78.00%和84.10%;转染后TGFβRⅠ干扰组的TGFβRⅠ蛋白表达量显著低于其阴性对照组;TGFβRⅡ干扰组、TGFβ1组和FSH组的TGFβRⅡ蛋白表达量显著低于其阴性对照组;TGFβ RI干扰组、TGFβ RII干扰组的增殖抑制率极显著高于PLL3.7空载体组,TGFβ 1组细胞增殖抑制率极显著高于空白对照组且FSH组的增殖抑制率显著高于空白对照组;TGFβRⅠ干扰组和TGFβRⅡ干扰组细胞凋亡率极显著高于PLL3.7空载体组;TGFβ1组和FSH组细胞凋亡率极显著高于空白对照组。总之,RNAi介导的TGFβRⅠ和TGFβRⅡ沉默和外源因子(10 ng/m L TGFβ1和25 ng/m L FSH)引起的TGFβRⅡ蛋白下调,均会减弱TGF-β信号传导通路对卵泡颗粒细胞的促进作用,进而抑制卵泡颗粒细胞的增殖,促进其凋亡。展开更多
Objective: To investigate the value of urinary trypsi-nogen activation peptide (TAP) in the early predic-tion of severe acute pancreatitis and to compare itwith acute physiology and chronic health evaluationⅡ (APACHE...Objective: To investigate the value of urinary trypsi-nogen activation peptide (TAP) in the early predic-tion of severe acute pancreatitis and to compare itwith acute physiology and chronic health evaluationⅡ (APACHE Ⅱ).Methods: We assessed the predictive value of urinaryTAP concentrations measured by a competitive en-zyme-linked immunosorbent assay. Urine sampleswere collected for detecting TAP concentrations atadmission, and 24, 48, and 72 h from 41 patientswith acute pancreatitis (12 with severe disease, 29with mild disease) who presented within 48 h the on-set of symptoms and from 11 control patients, whileAPACHE Ⅱ scores were recorded at 48 h after ad-mission.Results: The peak median urinary TAP concentrationwas seen at admission. The median urinary TAPconcentration at admission for severe pancreatitis (95nmol/L) was significantly higher than the medianfor patients with mild pancreatitis (20 nmol/L, P【0. 005) and controls (15 nmol/L, P【0. 005). TAPconcentrations were significantly higher in patientswith severe acute pancreatitis than the median in pa-tients with mild pancreatitis (P【0. 05) and controls(P【0. 05) on days 2 to 3. The median APACHE Ⅱscores of severe patients were significantly differentfrom those of mild patients (10.5 vs 6.0, P【0.01).The sensitivity, specificity, positive predictive, andnegative predictive values of an admission urinaryTAP≥35 nmol/L for severe pancreatitis were91.7%, 89.7%, 78.6% and 96.3%, whereas 48 hafter admission the values for APACHE Ⅱ scores(≥9) were 75.0%, 72.7%, 52.9% and 87.5%. Inprediction of disease severity, the urine TAP concen-tration was much better than APACHE Ⅱ at 48 h.Conclusions: Urinary TAP obtained at the first 48 hof the onset of symptoms can predict severe acutepancreatitis. In prediction of disease severity, theurinary TAP is much better than APACHE Ⅱ score.展开更多
目的:构建敲除TGF-βRⅡ的靶向CLDN18.2的CAR T细胞,并探索其在胃食管交界癌中的治疗潜力;方法:通过CRISPR/Cas9敲除T细胞中的TGF-βRⅡ,并使用慢病毒感染法构建CLDN18.2 CAR T细胞;流式细胞术检测CAR T细胞的阳性率、食管腺癌细胞系中C...目的:构建敲除TGF-βRⅡ的靶向CLDN18.2的CAR T细胞,并探索其在胃食管交界癌中的治疗潜力;方法:通过CRISPR/Cas9敲除T细胞中的TGF-βRⅡ,并使用慢病毒感染法构建CLDN18.2 CAR T细胞;流式细胞术检测CAR T细胞的阳性率、食管腺癌细胞系中CLDN18.2的表达情况、CAR T细胞中Smad2/3的磷酸化水平以及PD-1的表达水平;免疫荧光法检测CAR T细胞中TGF-βRⅡ的表达;LDH释放实验检测对靶细胞的细胞毒性;裸鼠移植瘤模型检测肿瘤抑制能力;ELISA实验检测肿瘤组织中细胞因子的释放水平;结果:成功构建敲除TGF-βRⅡ的CLDN18.2 CAR T细胞(18.2BBZ/TGFBRKO)以及二代CLDN18.2 CAR T细胞(CLDN18.2);18.2BBZ及18.2BBZ/TGFBRKO在体外对CLDN18.2+的肿瘤细胞具有剂量依赖型细胞毒性;TGF-β1不能使18.2BBZ/TGFBRKO的Smad2/3发生磷酸化,在TGF-β1存在时,18.2BBZ/TGFBRKO与肿瘤细胞共孵育后PD-1的表达水平显著低于18.2BBZ(P<0.001);18.2BBZ/TGFBRKO较18.2BBZ对裸鼠移植瘤的抑制能力更强(P<0.0001),并且在肿瘤组织中保持更强的细胞因子分泌水平(P<0.01);结论:通过CRISPR/Cas9敲除CLDN18.2 CAR T细胞中的TGF-βRⅡ可以有效降低T细胞耗竭,并在小鼠移植瘤模型中产生更强的抑制肿瘤活性,并增加促炎性细胞因子的分泌。展开更多
AIM: To undertake a meta-analysis on the value of urinary trypsinogen activation peptide (uTAP) in predicting severity of acute pancreatitis on admission.METHODS: Major databases including Medline, Embase, Science Cit...AIM: To undertake a meta-analysis on the value of urinary trypsinogen activation peptide (uTAP) in predicting severity of acute pancreatitis on admission.METHODS: Major databases including Medline, Embase, Science Citation Index Expanded and the Cochrane Central Register of Controlled Trials in the Cochrane Library were searched to identify all relevant studies from January 1990 to January 2013. Pooled sensitivity, specificity and the diagnostic odds ratios (DORs) with 95%CI were calculated for each study and were compared to other systems/biomarkers if mentioned within the same study. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated.RESULTS: In total, six studies of uTAP with a cut-off value of 35 nmol/L were included in this meta-analysis. Overall, the pooled sensitivity and specificity of uTAP for predicting severity of acute pancreatitis, at time of admission, was 71% and 75%, respectively (AUC = 0.83, DOR = 8.67, 95%CI: 3.70-20.33). When uTAP was compared with plasma C-reactive protein, the pooled sensitivity, specificity, AUC and DOR were 0.64 vs 0.67, 0.77 vs 0.75, 0.82 vs 0.79 and 6.27 vs 6.32, respectively. Similarly, the pooled sensitivity, specificity, AUC and DOR of uTAP vs Acute Physiology and Chronic Health Evaluation II within the first 48 h of admission were found to be 0.64 vs 0.69, 0.77 vs 0.61, 0.82 vs 0.73 and 6.27 vs 4.61, respectively.CONCLUSION: uTAP has the potential to act as a stratification marker on admission for differentiating disease severity of acute pancreatitis.展开更多
The interaction among type Ⅱ collagen(CⅡ),human DR4 major histocompatibility complex type Ⅱ molecule(MHC Ⅱ)and T-cell receptor(TCR)is associated with the development of rheumatoid arthritis(RA).The activation of T...The interaction among type Ⅱ collagen(CⅡ),human DR4 major histocompatibility complex type Ⅱ molecule(MHC Ⅱ)and T-cell receptor(TCR)is associated with the development of rheumatoid arthritis(RA).The activation of T cells can be reduced through exposure to modified CⅡ(263-272)glycopeptide fragment via competitive inhibition with self-antigen.In this work,30 peptides based on the sequence of CⅡ(263-272)were prepared and evaluated for their binding to DR4 protein by surface plasmon resonance(SPR)assay.The effect on the secretion of pro-inflammatory factors by the spleen cells in collagen induced rheumatoid arthritis(CIA)mouse was also investigated.Two N-glycosylated CⅡ peptides were identified to have strong binding to the human recombinant DR4 protein and weak proinflammatory effect.These glycopeptides could be developed as therapeutic saccharide vaccines for the treatment of rheumatoid arthritis(RA).展开更多
基金This work was supported by the National Natural Science Foundation of China(Nos.30470408 and 20637010).
文摘The inhibitory mechanism of copper（Ⅱ） on the aggegation of amyloid β-peptide （Aβ） was investigated by molecular dynamics simulations. The binding mode ofcopper（Ⅱ） with Aβ is characterized by the imidazole nitrogen atom, Nπ, of the histidine residue H 13, acting as the anchoring site, and the backbone＇s deprotoned amide nitogen atoms as the main binding sites. Drove by the coordination bonds and their induced hydrogen bond net, the conformations of Aβ converted from β-sheet non-β-sheet conformations, which destabilized the aggregation of Aβ into fibrils.
文摘为研究TGF-β通路对猪颗粒细胞增殖、凋亡的影响,本研究对TGF-β通路基因进行调控处理,设计并筛选可高效干扰TGFβRⅠ及TGFβRⅡ表达的载体,转染不同干扰载体及添加10 ng/m L TGFβ1和25 ng/m L FSH,采用Westernblot、MTT和流式细胞法检测不同处理对颗粒细胞TGFβ RI和TGFβ RII蛋白表达量和细胞增殖凋亡的影响。结果表明:成功筛选出干扰效率最高的p OSP1-TGFβRⅠ-sh5和p OSP1-TGFβRⅡ-sh3载体,对TGFβRⅠ和TGFβRⅡ的抑制率分别为78.00%和84.10%;转染后TGFβRⅠ干扰组的TGFβRⅠ蛋白表达量显著低于其阴性对照组;TGFβRⅡ干扰组、TGFβ1组和FSH组的TGFβRⅡ蛋白表达量显著低于其阴性对照组;TGFβ RI干扰组、TGFβ RII干扰组的增殖抑制率极显著高于PLL3.7空载体组,TGFβ 1组细胞增殖抑制率极显著高于空白对照组且FSH组的增殖抑制率显著高于空白对照组;TGFβRⅠ干扰组和TGFβRⅡ干扰组细胞凋亡率极显著高于PLL3.7空载体组;TGFβ1组和FSH组细胞凋亡率极显著高于空白对照组。总之,RNAi介导的TGFβRⅠ和TGFβRⅡ沉默和外源因子(10 ng/m L TGFβ1和25 ng/m L FSH)引起的TGFβRⅡ蛋白下调,均会减弱TGF-β信号传导通路对卵泡颗粒细胞的促进作用,进而抑制卵泡颗粒细胞的增殖,促进其凋亡。
文摘Objective: To investigate the value of urinary trypsi-nogen activation peptide (TAP) in the early predic-tion of severe acute pancreatitis and to compare itwith acute physiology and chronic health evaluationⅡ (APACHE Ⅱ).Methods: We assessed the predictive value of urinaryTAP concentrations measured by a competitive en-zyme-linked immunosorbent assay. Urine sampleswere collected for detecting TAP concentrations atadmission, and 24, 48, and 72 h from 41 patientswith acute pancreatitis (12 with severe disease, 29with mild disease) who presented within 48 h the on-set of symptoms and from 11 control patients, whileAPACHE Ⅱ scores were recorded at 48 h after ad-mission.Results: The peak median urinary TAP concentrationwas seen at admission. The median urinary TAPconcentration at admission for severe pancreatitis (95nmol/L) was significantly higher than the medianfor patients with mild pancreatitis (20 nmol/L, P【0. 005) and controls (15 nmol/L, P【0. 005). TAPconcentrations were significantly higher in patientswith severe acute pancreatitis than the median in pa-tients with mild pancreatitis (P【0. 05) and controls(P【0. 05) on days 2 to 3. The median APACHE Ⅱscores of severe patients were significantly differentfrom those of mild patients (10.5 vs 6.0, P【0.01).The sensitivity, specificity, positive predictive, andnegative predictive values of an admission urinaryTAP≥35 nmol/L for severe pancreatitis were91.7%, 89.7%, 78.6% and 96.3%, whereas 48 hafter admission the values for APACHE Ⅱ scores(≥9) were 75.0%, 72.7%, 52.9% and 87.5%. Inprediction of disease severity, the urine TAP concen-tration was much better than APACHE Ⅱ at 48 h.Conclusions: Urinary TAP obtained at the first 48 hof the onset of symptoms can predict severe acutepancreatitis. In prediction of disease severity, theurinary TAP is much better than APACHE Ⅱ score.
文摘目的:构建敲除TGF-βRⅡ的靶向CLDN18.2的CAR T细胞,并探索其在胃食管交界癌中的治疗潜力;方法:通过CRISPR/Cas9敲除T细胞中的TGF-βRⅡ,并使用慢病毒感染法构建CLDN18.2 CAR T细胞;流式细胞术检测CAR T细胞的阳性率、食管腺癌细胞系中CLDN18.2的表达情况、CAR T细胞中Smad2/3的磷酸化水平以及PD-1的表达水平;免疫荧光法检测CAR T细胞中TGF-βRⅡ的表达;LDH释放实验检测对靶细胞的细胞毒性;裸鼠移植瘤模型检测肿瘤抑制能力;ELISA实验检测肿瘤组织中细胞因子的释放水平;结果:成功构建敲除TGF-βRⅡ的CLDN18.2 CAR T细胞(18.2BBZ/TGFBRKO)以及二代CLDN18.2 CAR T细胞(CLDN18.2);18.2BBZ及18.2BBZ/TGFBRKO在体外对CLDN18.2+的肿瘤细胞具有剂量依赖型细胞毒性;TGF-β1不能使18.2BBZ/TGFBRKO的Smad2/3发生磷酸化,在TGF-β1存在时,18.2BBZ/TGFBRKO与肿瘤细胞共孵育后PD-1的表达水平显著低于18.2BBZ(P<0.001);18.2BBZ/TGFBRKO较18.2BBZ对裸鼠移植瘤的抑制能力更强(P<0.0001),并且在肿瘤组织中保持更强的细胞因子分泌水平(P<0.01);结论:通过CRISPR/Cas9敲除CLDN18.2 CAR T细胞中的TGF-βRⅡ可以有效降低T细胞耗竭,并在小鼠移植瘤模型中产生更强的抑制肿瘤活性,并增加促炎性细胞因子的分泌。
基金Supported by Technology Supported Program of Sichuan Province, No. 2011SZ0291the National Natural Science Foundation of China, No. 81072910National Institute for Health Research, United Kingdom
文摘AIM: To undertake a meta-analysis on the value of urinary trypsinogen activation peptide (uTAP) in predicting severity of acute pancreatitis on admission.METHODS: Major databases including Medline, Embase, Science Citation Index Expanded and the Cochrane Central Register of Controlled Trials in the Cochrane Library were searched to identify all relevant studies from January 1990 to January 2013. Pooled sensitivity, specificity and the diagnostic odds ratios (DORs) with 95%CI were calculated for each study and were compared to other systems/biomarkers if mentioned within the same study. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated.RESULTS: In total, six studies of uTAP with a cut-off value of 35 nmol/L were included in this meta-analysis. Overall, the pooled sensitivity and specificity of uTAP for predicting severity of acute pancreatitis, at time of admission, was 71% and 75%, respectively (AUC = 0.83, DOR = 8.67, 95%CI: 3.70-20.33). When uTAP was compared with plasma C-reactive protein, the pooled sensitivity, specificity, AUC and DOR were 0.64 vs 0.67, 0.77 vs 0.75, 0.82 vs 0.79 and 6.27 vs 6.32, respectively. Similarly, the pooled sensitivity, specificity, AUC and DOR of uTAP vs Acute Physiology and Chronic Health Evaluation II within the first 48 h of admission were found to be 0.64 vs 0.69, 0.77 vs 0.61, 0.82 vs 0.73 and 6.27 vs 4.61, respectively.CONCLUSION: uTAP has the potential to act as a stratification marker on admission for differentiating disease severity of acute pancreatitis.
基金supported by the National Natural Science Foundation of China(Nos.81930097,21977005,82151223)supported by the National Key R&D Program of China(No.2022YFF1203005).
文摘The interaction among type Ⅱ collagen(CⅡ),human DR4 major histocompatibility complex type Ⅱ molecule(MHC Ⅱ)and T-cell receptor(TCR)is associated with the development of rheumatoid arthritis(RA).The activation of T cells can be reduced through exposure to modified CⅡ(263-272)glycopeptide fragment via competitive inhibition with self-antigen.In this work,30 peptides based on the sequence of CⅡ(263-272)were prepared and evaluated for their binding to DR4 protein by surface plasmon resonance(SPR)assay.The effect on the secretion of pro-inflammatory factors by the spleen cells in collagen induced rheumatoid arthritis(CIA)mouse was also investigated.Two N-glycosylated CⅡ peptides were identified to have strong binding to the human recombinant DR4 protein and weak proinflammatory effect.These glycopeptides could be developed as therapeutic saccharide vaccines for the treatment of rheumatoid arthritis(RA).
