AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided ...AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided into two groups:ethanol-fed group (group E) and control group (group C). Group E was fed with ethanol(5-12 g x kg(-1) x d(-1)) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 weeks and 8 weeks. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed under light and electronic microscopy. Expressions of LBP and CD14 mRNA in liver tissues were determined by RT-PCR analysis. RESULTS: Plasma endotoxin levels were increased more significantly in group E(129+/-21) ng x L(-1) and (187+/-35) ng x L(-1) at 4 and 8 wk than in control rats(48+/-9) ng x L(-1) and (53+/-11) ng x L(-1), respectively (P【0.05). Mean values of plasma ALT levels were (1867+/-250) nkat x L(-1) and (2450+/-367) nkat x L(-1) in Group E. The values were increased more dramatically in ethanol-fed rats than in Group C after 4 and 8 weeks. In liver section from ethanol-fed rats, there were marked pathological changes (steatosis, cell infiltration and necrosis). In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels compared with the control group (P【0.05). CONCLUSION: Ethanol administration led to a significant increase in endotoxin levels in serum and LBP and CD14 mRNA expressions in liver tissues. The increase of LBP and CD14 mRNA expression might wake the liver more sensitive to endotoxin and liver injury.展开更多
In this paper we aim to analyse temporal variation of CD4 cell counts for HIV-infected individuals under antiretroviral therapy by using statistical methods. This is achieved by resorting to recursive binary regressio...In this paper we aim to analyse temporal variation of CD4 cell counts for HIV-infected individuals under antiretroviral therapy by using statistical methods. This is achieved by resorting to recursive binary regression tree approach [1]?[2]. This approach has made it possible to highlight the existence of several segments of the population of interest described by the interactions between the predictive covariates of the response to the treatment regimen.展开更多
目的:构建ZCH-7-2F9(简称2F9)单链抗体(ScFv2F9)原核表达载体,获得活性目的蛋白,为2F9免疫毒素及其它类型的基因工程抗体的研究奠定基础。方法:根据VH2F9、VL2F9、(G4S)3基因序列以及pIVEX2.3-MCS空载体多克隆位点内切酶位点NdeI和SmaI...目的:构建ZCH-7-2F9(简称2F9)单链抗体(ScFv2F9)原核表达载体,获得活性目的蛋白,为2F9免疫毒素及其它类型的基因工程抗体的研究奠定基础。方法:根据VH2F9、VL2F9、(G4S)3基因序列以及pIVEX2.3-MCS空载体多克隆位点内切酶位点NdeI和SmaI的核酸序列设计引物,采用高保真的Taq酶,通过重叠延伸拚接法(splicing by overlap extension,SOE)扩增克隆到ScFv2F9基因,T-A克隆、测序核实后克隆到pIVEX2.3-MCS空载体中,阳性重组质粒转化大肠杆菌菌株E.coli BL21star(DE3)plysS,IPTG诱导目的蛋白表达,镍树脂纯化后体外复性浓缩,流式细胞术观察其识别和结合CD14的能力。结果:成功构建ScFv2F9的原核表达载体pIVEX2.3-MCS/ScFv2F9;对大肠杆菌的包涵体进行纯化复性,其复性率为32.6%;通过流式细胞术观察到复性ScFv2F9对亲本单抗2F9-FITC有部分的阻滞效果,复性ScFv2F9阻滞1次后阳性细胞百分数、平均荧光强度(MFI)和高峰频道(peakCh)分别下降了11.73%、11.96%和31.57%,阻滞2次后分别下降了26.44%、21.75%和42.11%。结论:ScFv2F9在原核细胞中获得成功表达,复性后目的蛋白对人CD14有一定的识别和结合能力,为2F9单抗的进一步改造打下了坚实的基础。展开更多
BACKGROUND Colorectal cancer(CRC)remains one of the leading causes of cancer-related morbidity and mortality worldwide.Growing evidence suggests that gut microbial dysbiosis plays a crucial role in tumorigenesis and c...BACKGROUND Colorectal cancer(CRC)remains one of the leading causes of cancer-related morbidity and mortality worldwide.Growing evidence suggests that gut microbial dysbiosis plays a crucial role in tumorigenesis and can influence therapeutic responses.AIM To explore the associations between serum S100A12 and soluble CD14(sCD14)levels and gut microbiota alterations in patients with CRC,and to assess the predictive utility of these biomarkers in forecasting chemotherapy response.METHODS A retrospective analysis was conducted on 104 patients diagnosed with advanced CRC(CRC group)and 104 age-matched and sex-matched healthy controls.Serum concentrations of S100A12 and sCD14 were measured using enzyme-linked immunosorbent assay.Fecal samples collected before chemotherapy were subjected to 16S rRNA sequencing to profile gut microbial composition.Pearson correlation analysis was used to evaluate the relationship between biomarker levels and microbial abundance.Receiver operating characteristic(ROC)curves were used to assess the predictive performance of S100A12 and sCD14 for chemotherapy response.RESULTS CRC patients exhibited significantly higher serum levels of S100A12 and sCD14 compared to healthy individuals(P<0.05).Patients with moderate to severe gut dysbiosis showed the highest elevations of these biomarkers(P<0.05).Elevated levels of S100A12 and sCD14 were positively correlated with Fusobacterium nucleatum and Prevotella abundance,and negatively correlated with Faecalibacterium prausnitzii and Akkermansia muciniphila(P<0.05).Both biomarkers significantly decreased following chemotherapy(P<0.05).Non-responders to chemotherapy had higher pre-treatment levels of S100A12 and sCD14 compared to responders(P<0.05).Combined ROC analysis showed improved diagnostic accuracy compared to either marker alone.CONCLUSION Serum S100A12 and sCD14 levels are closely associated with gut microbiota imbalance and chemotherapy response in CRC patients.These markers may serve as promising predictive indicators for treatment efficacy and offer potential value in individualized treatment strategies.展开更多
基金Supported by the National Natural Science Foundation of China(No.39970719).
文摘AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided into two groups:ethanol-fed group (group E) and control group (group C). Group E was fed with ethanol(5-12 g x kg(-1) x d(-1)) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 weeks and 8 weeks. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed under light and electronic microscopy. Expressions of LBP and CD14 mRNA in liver tissues were determined by RT-PCR analysis. RESULTS: Plasma endotoxin levels were increased more significantly in group E(129+/-21) ng x L(-1) and (187+/-35) ng x L(-1) at 4 and 8 wk than in control rats(48+/-9) ng x L(-1) and (53+/-11) ng x L(-1), respectively (P【0.05). Mean values of plasma ALT levels were (1867+/-250) nkat x L(-1) and (2450+/-367) nkat x L(-1) in Group E. The values were increased more dramatically in ethanol-fed rats than in Group C after 4 and 8 weeks. In liver section from ethanol-fed rats, there were marked pathological changes (steatosis, cell infiltration and necrosis). In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels compared with the control group (P【0.05). CONCLUSION: Ethanol administration led to a significant increase in endotoxin levels in serum and LBP and CD14 mRNA expressions in liver tissues. The increase of LBP and CD14 mRNA expression might wake the liver more sensitive to endotoxin and liver injury.
文摘In this paper we aim to analyse temporal variation of CD4 cell counts for HIV-infected individuals under antiretroviral therapy by using statistical methods. This is achieved by resorting to recursive binary regression tree approach [1]?[2]. This approach has made it possible to highlight the existence of several segments of the population of interest described by the interactions between the predictive covariates of the response to the treatment regimen.
文摘目的:构建ZCH-7-2F9(简称2F9)单链抗体(ScFv2F9)原核表达载体,获得活性目的蛋白,为2F9免疫毒素及其它类型的基因工程抗体的研究奠定基础。方法:根据VH2F9、VL2F9、(G4S)3基因序列以及pIVEX2.3-MCS空载体多克隆位点内切酶位点NdeI和SmaI的核酸序列设计引物,采用高保真的Taq酶,通过重叠延伸拚接法(splicing by overlap extension,SOE)扩增克隆到ScFv2F9基因,T-A克隆、测序核实后克隆到pIVEX2.3-MCS空载体中,阳性重组质粒转化大肠杆菌菌株E.coli BL21star(DE3)plysS,IPTG诱导目的蛋白表达,镍树脂纯化后体外复性浓缩,流式细胞术观察其识别和结合CD14的能力。结果:成功构建ScFv2F9的原核表达载体pIVEX2.3-MCS/ScFv2F9;对大肠杆菌的包涵体进行纯化复性,其复性率为32.6%;通过流式细胞术观察到复性ScFv2F9对亲本单抗2F9-FITC有部分的阻滞效果,复性ScFv2F9阻滞1次后阳性细胞百分数、平均荧光强度(MFI)和高峰频道(peakCh)分别下降了11.73%、11.96%和31.57%,阻滞2次后分别下降了26.44%、21.75%和42.11%。结论:ScFv2F9在原核细胞中获得成功表达,复性后目的蛋白对人CD14有一定的识别和结合能力,为2F9单抗的进一步改造打下了坚实的基础。
文摘BACKGROUND Colorectal cancer(CRC)remains one of the leading causes of cancer-related morbidity and mortality worldwide.Growing evidence suggests that gut microbial dysbiosis plays a crucial role in tumorigenesis and can influence therapeutic responses.AIM To explore the associations between serum S100A12 and soluble CD14(sCD14)levels and gut microbiota alterations in patients with CRC,and to assess the predictive utility of these biomarkers in forecasting chemotherapy response.METHODS A retrospective analysis was conducted on 104 patients diagnosed with advanced CRC(CRC group)and 104 age-matched and sex-matched healthy controls.Serum concentrations of S100A12 and sCD14 were measured using enzyme-linked immunosorbent assay.Fecal samples collected before chemotherapy were subjected to 16S rRNA sequencing to profile gut microbial composition.Pearson correlation analysis was used to evaluate the relationship between biomarker levels and microbial abundance.Receiver operating characteristic(ROC)curves were used to assess the predictive performance of S100A12 and sCD14 for chemotherapy response.RESULTS CRC patients exhibited significantly higher serum levels of S100A12 and sCD14 compared to healthy individuals(P<0.05).Patients with moderate to severe gut dysbiosis showed the highest elevations of these biomarkers(P<0.05).Elevated levels of S100A12 and sCD14 were positively correlated with Fusobacterium nucleatum and Prevotella abundance,and negatively correlated with Faecalibacterium prausnitzii and Akkermansia muciniphila(P<0.05).Both biomarkers significantly decreased following chemotherapy(P<0.05).Non-responders to chemotherapy had higher pre-treatment levels of S100A12 and sCD14 compared to responders(P<0.05).Combined ROC analysis showed improved diagnostic accuracy compared to either marker alone.CONCLUSION Serum S100A12 and sCD14 levels are closely associated with gut microbiota imbalance and chemotherapy response in CRC patients.These markers may serve as promising predictive indicators for treatment efficacy and offer potential value in individualized treatment strategies.
