目的探究血清S100钙结合蛋白A8(S100A8)和程序性死亡配体-1(PD-L1)联合超声检测在子宫内膜癌诊断及预后中的价值。方法选取2020年4月至2022年4月于我院收治的确诊为子宫内膜癌的患者166例即为癌症组,按照国际妇产科联合会(FIGO)标准分...目的探究血清S100钙结合蛋白A8(S100A8)和程序性死亡配体-1(PD-L1)联合超声检测在子宫内膜癌诊断及预后中的价值。方法选取2020年4月至2022年4月于我院收治的确诊为子宫内膜癌的患者166例即为癌症组,按照国际妇产科联合会(FIGO)标准分为Ⅰ期48例、Ⅱ期52例、Ⅲ期42例、Ⅳ期24例。选择同期在我院确诊为子宫内膜增生的患者166例为对照组;使用酶联免疫吸附试验(ELISA)检测血清中S100A8和PD-L1水平;使用阴道超声获取血流频谱测得血流参数:搏动指数(PI)、阻力指数(RI)。采用受试者工作特征(ROC)曲线分析血清S100A8和PD-L1联合超声参数对子宫内膜癌的诊断价值以及预后的预测价值。结果与对照组相比,癌症组患者血清中S100A8和PD-L1水平显著升高(t=10.688、8.605,均P<0.001);与Ⅰ期相比,Ⅱ期、Ⅲ期、Ⅳ期患者血清中S100A8和PD-L1水平显著升高,与Ⅱ期相比,Ⅳ期患者血清中S100A8和PD-L1水平显著升高(P<0.05)。与对照组相比,癌症组PI和RI的值显著降低(t=7.183、6.799,均P<0.001);血清S100A8和PD-L1联合超声参数对子宫内膜癌诊断的曲线下面积(AUC)高于各指标单独诊断的AUC值(Z S100A8 vs.S100A8+PD-L1+PI+RI=6.472,Z PD-L1 vs.S100A8+PD-L1+PI+RI=6.903,Z PI vs.S100A8+PD-L1+PI+RI=7.072,Z RI vs.S100A8+PD-L1+PI+RI=5.987,均P<0.001);与预后良好组相比,预后不良组S100A8、PD-L1水平显著升高,PI、RI水平显著降低(P<0.001);血清S100A8和PD-L1联合超声参数对子宫内膜癌预后预测的AUC高于各指标单独预测的AUC值(Z S100A8 vs.S100A8+PD-L1+PI+RI=2.841,P=0.005;Z PD-L1 vs.S100A8+PD-L1+PI+RI=2.146,P=0.032;Z PI vs.S100A8+PD-L1+PI+RI=6.056,P<0.001;Z RI vs.S100A8+PD-L1+PI+RI=4.370,P<0.001)。结论子宫内膜癌患者血清中S100A8和PD-L1水平显著升高,血清S100A8和PD-L1联合超声能够提高对子宫内膜癌诊断及预后价值。展开更多
Quantitative analysis of dendritic cells (DC’s) was carried out in tissue specimens of normalgastric mucosa (n=15),gastric ulcer (n=19),chronic atrophic gastritis (n=28),and gastriccarcinoma (n=65) by ABC immunostain...Quantitative analysis of dendritic cells (DC’s) was carried out in tissue specimens of normalgastric mucosa (n=15),gastric ulcer (n=19),chronic atrophic gastritis (n=28),and gastriccarcinoma (n=65) by ABC immunostaining with S100 protein antibody.Significant increasein DC number were observed in chronic atrophic gastritis with type Ⅲ intestinal metaplasiaand/or grade Ⅱ,Ⅲ dysplasia.The result suggests that DC’s are potentially capable opresenting neoantigens associated with malignant transformation at the precancerous stagewhen malignant morphological changes have not yet taken place.Combined with routinediagnostic methods,the serial monitoring of DC density in gastric mucosa may be usefulin the follow-up of premalignant lesions in the stomach and the diagnosis of early gastriccarcinoma.展开更多
BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural...BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.展开更多
BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mech...BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mechanisms of traditional Chinese medicine against ERS in AD are poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. NC was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yinxingye （Ginkgo Leaf） in a proportion of 1 ： 2： 2. Following conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.展开更多
Objective: To investigate the effects of Propofol combined with remifentanil on serum levels of MBP, NSE and S100B protein, D-D and inflammatory factors in patients with acute craniocerebral trauma. Methods: A total o...Objective: To investigate the effects of Propofol combined with remifentanil on serum levels of MBP, NSE and S100B protein, D-D and inflammatory factors in patients with acute craniocerebral trauma. Methods: A total of 100 patients were selected with traumatic brain injury who underwent emergency surgery from August 2014 to May 2017 in our hospital, then randomly divided them into the control group and the experimental group, 50 cases each. The control group received isoflurane combined with remifentanil to maintain anesthesia, and the experimental group received propofol and remifentanil to maintain anesthesia. The inflammatory factors and the levels of MBP, NSE, S100B and D-D in the two groups before and after anesthesia (T0), 1H (T1) and postoperative 1H (T2) were detected and compared. Results: There was no significant difference between the two groups in the levels of TNF-α. The serum level of hs-CRP in two groups of T1, T2 increased significantly, the difference was statistically significant compared with T0, in the experimental group, serum level of hs-CRP at T1 and T2 was significantly higher than the control group, the difference was statistically significant. Conclusion: Propofol combined with remifentanil anesthesia for acute craniocerebral trauma can maintain the balance of inflammatory cytokine levels during the perioperative period, inhibit the elevation of serum MBP, NSE, S100B protein and D-D levels, reduce brain cell damage. It has a good protective effect on brain cells and is worthy of clinical application.展开更多
Objective To explore the effect of β-amyloid protein (Aβ) on S100β expression in rat hippocampus and its mechanisms. Methods At 7 days after bilateral stereotaxis injection of different dose of fibrillar Aβ 25-35 ...Objective To explore the effect of β-amyloid protein (Aβ) on S100β expression in rat hippocampus and its mechanisms. Methods At 7 days after bilateral stereotaxis injection of different dose of fibrillar Aβ 25-35 and interluekin-1 receptor antagonist (IL-1ra) into the rat CA1 region, the learning and memory abilities of rats were tested with passive avoidance task. Amyloid deposition was detected by using Congo red staining technique. Nissl staining and immunohistochemical techniques were used to analyze the number of neurons, and GFAP and the S100β expression in hippocampal CA1 region , respectively. Results After fibrillar Aβ injection, the step-through latency of rats was significantly shortened compared to that of the control group. The GFAP positive astrocytes were found surrounding amyloid deposition. Neuronal loss occurred in the pyramidal cell layer of CA1 region. The number of S100β positive cells in Aβ-treated group was significantly increased compared with that in the control group. After IL-1ra injection, the number of S100β positive cells was significantly decreased. Conclusion Intrahippocampal injection of Aβ 25-35 could cause similar pathologic changes of Alzheimer’s disease. Aβ 25-35 was capable of up-regulating S100β expression in a dose-dependent manner. The injection of IL-1ra could attenuate the effect of Aβ on S100β expression.展开更多
The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative di...The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative diseases. In the present study, we observed the effects of transfecting three amyloid precursor-like protein 2 （APLP2） C-terminal fragments （CTFs; C57, C50 and C31） in the pEGFP-N1 vector on S100A9 expression in BV2 cells. Reverse transcription-PCR, western blot assay and immunocytochemistry revealed that S100A9 protein and mRNA expression was greater in BV2 cells after CTF transfection than after mock transfection with an empty vector. Furthermore, transfection of full-length APLP2-751 resulted in low levels of S100A9 protein ex-pression. Our results show that APLP2-CTFs upregulate S100A9 protein and mRNA expression in BV2 cells, and identify a novel pathway involved in neuronal injury and apoptosis, and repair and protection in Alzheimer’s disease.展开更多
The ability to visualise proteins in their native environment and discern information regarding stoichiometry is of critical importance when studying protein interactions and function. We have used liquid cell atomic ...The ability to visualise proteins in their native environment and discern information regarding stoichiometry is of critical importance when studying protein interactions and function. We have used liquid cell atomic force microscopy (AFM) to visualise proteins in their native state in buffer and have determined their molecular volumes. The human proteins S100A8, S100A9, S100A12 and CLIC1 were used in this investigation. The effect of oxidation on the protein structure of CLIC1 was also investigated and we found that CLIC1 multimerisation could be discerned by AFM, which supports similar findings by other methods. We have found good correlation between the molecular volumes measured by AFM and the calculated volumes of the individual proteins. This method allows for the study of single soluble proteins under physiological conditions and could potentially be extended to study the structure of these proteins when located within a membrane environment.展开更多
文摘目的探究血清S100钙结合蛋白A8(S100A8)和程序性死亡配体-1(PD-L1)联合超声检测在子宫内膜癌诊断及预后中的价值。方法选取2020年4月至2022年4月于我院收治的确诊为子宫内膜癌的患者166例即为癌症组,按照国际妇产科联合会(FIGO)标准分为Ⅰ期48例、Ⅱ期52例、Ⅲ期42例、Ⅳ期24例。选择同期在我院确诊为子宫内膜增生的患者166例为对照组;使用酶联免疫吸附试验(ELISA)检测血清中S100A8和PD-L1水平;使用阴道超声获取血流频谱测得血流参数:搏动指数(PI)、阻力指数(RI)。采用受试者工作特征(ROC)曲线分析血清S100A8和PD-L1联合超声参数对子宫内膜癌的诊断价值以及预后的预测价值。结果与对照组相比,癌症组患者血清中S100A8和PD-L1水平显著升高(t=10.688、8.605,均P<0.001);与Ⅰ期相比,Ⅱ期、Ⅲ期、Ⅳ期患者血清中S100A8和PD-L1水平显著升高,与Ⅱ期相比,Ⅳ期患者血清中S100A8和PD-L1水平显著升高(P<0.05)。与对照组相比,癌症组PI和RI的值显著降低(t=7.183、6.799,均P<0.001);血清S100A8和PD-L1联合超声参数对子宫内膜癌诊断的曲线下面积(AUC)高于各指标单独诊断的AUC值(Z S100A8 vs.S100A8+PD-L1+PI+RI=6.472,Z PD-L1 vs.S100A8+PD-L1+PI+RI=6.903,Z PI vs.S100A8+PD-L1+PI+RI=7.072,Z RI vs.S100A8+PD-L1+PI+RI=5.987,均P<0.001);与预后良好组相比,预后不良组S100A8、PD-L1水平显著升高,PI、RI水平显著降低(P<0.001);血清S100A8和PD-L1联合超声参数对子宫内膜癌预后预测的AUC高于各指标单独预测的AUC值(Z S100A8 vs.S100A8+PD-L1+PI+RI=2.841,P=0.005;Z PD-L1 vs.S100A8+PD-L1+PI+RI=2.146,P=0.032;Z PI vs.S100A8+PD-L1+PI+RI=6.056,P<0.001;Z RI vs.S100A8+PD-L1+PI+RI=4.370,P<0.001)。结论子宫内膜癌患者血清中S100A8和PD-L1水平显著升高,血清S100A8和PD-L1联合超声能够提高对子宫内膜癌诊断及预后价值。
文摘Quantitative analysis of dendritic cells (DC’s) was carried out in tissue specimens of normalgastric mucosa (n=15),gastric ulcer (n=19),chronic atrophic gastritis (n=28),and gastriccarcinoma (n=65) by ABC immunostaining with S100 protein antibody.Significant increasein DC number were observed in chronic atrophic gastritis with type Ⅲ intestinal metaplasiaand/or grade Ⅱ,Ⅲ dysplasia.The result suggests that DC’s are potentially capable opresenting neoantigens associated with malignant transformation at the precancerous stagewhen malignant morphological changes have not yet taken place.Combined with routinediagnostic methods,the serial monitoring of DC density in gastric mucosa may be usefulin the follow-up of premalignant lesions in the stomach and the diagnosis of early gastriccarcinoma.
基金the National High-Tech Research & Development Program of China, No. 2006AA02A128the National Natural Science Foundation of China, No. 30670667
文摘BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.
基金the National Natural Science Foundation of China, No. 30973779the National Special Planning Project for Traditional Chinese Medicine of China, No.02-03LP41the Key Program of Scientific Planning of Guangdong Province, No. 2006B35630007
文摘BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mechanisms of traditional Chinese medicine against ERS in AD are poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. NC was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yinxingye （Ginkgo Leaf） in a proportion of 1 ： 2： 2. Following conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.
基金The Natural Science Foundation of Shaanxi Province(2016JQ2341).
文摘Objective: To investigate the effects of Propofol combined with remifentanil on serum levels of MBP, NSE and S100B protein, D-D and inflammatory factors in patients with acute craniocerebral trauma. Methods: A total of 100 patients were selected with traumatic brain injury who underwent emergency surgery from August 2014 to May 2017 in our hospital, then randomly divided them into the control group and the experimental group, 50 cases each. The control group received isoflurane combined with remifentanil to maintain anesthesia, and the experimental group received propofol and remifentanil to maintain anesthesia. The inflammatory factors and the levels of MBP, NSE, S100B and D-D in the two groups before and after anesthesia (T0), 1H (T1) and postoperative 1H (T2) were detected and compared. Results: There was no significant difference between the two groups in the levels of TNF-α. The serum level of hs-CRP in two groups of T1, T2 increased significantly, the difference was statistically significant compared with T0, in the experimental group, serum level of hs-CRP at T1 and T2 was significantly higher than the control group, the difference was statistically significant. Conclusion: Propofol combined with remifentanil anesthesia for acute craniocerebral trauma can maintain the balance of inflammatory cytokine levels during the perioperative period, inhibit the elevation of serum MBP, NSE, S100B protein and D-D levels, reduce brain cell damage. It has a good protective effect on brain cells and is worthy of clinical application.
文摘Objective To explore the effect of β-amyloid protein (Aβ) on S100β expression in rat hippocampus and its mechanisms. Methods At 7 days after bilateral stereotaxis injection of different dose of fibrillar Aβ 25-35 and interluekin-1 receptor antagonist (IL-1ra) into the rat CA1 region, the learning and memory abilities of rats were tested with passive avoidance task. Amyloid deposition was detected by using Congo red staining technique. Nissl staining and immunohistochemical techniques were used to analyze the number of neurons, and GFAP and the S100β expression in hippocampal CA1 region , respectively. Results After fibrillar Aβ injection, the step-through latency of rats was significantly shortened compared to that of the control group. The GFAP positive astrocytes were found surrounding amyloid deposition. Neuronal loss occurred in the pyramidal cell layer of CA1 region. The number of S100β positive cells in Aβ-treated group was significantly increased compared with that in the control group. After IL-1ra injection, the number of S100β positive cells was significantly decreased. Conclusion Intrahippocampal injection of Aβ 25-35 could cause similar pathologic changes of Alzheimer’s disease. Aβ 25-35 was capable of up-regulating S100β expression in a dose-dependent manner. The injection of IL-1ra could attenuate the effect of Aβ on S100β expression.
基金supported by the Natural Science Foundation of Technology Gallery in Jilin Province of China,No.2011-15237the National Natural Science Foundation of China,No.81160159
文摘The murine microglial cell line BV2 has neuroprotective effects, but is toxic to neurons by secret-ing inlfammatory cytokines, and is an important target in the treatment of nerve inlfammation and neurodegenerative diseases. In the present study, we observed the effects of transfecting three amyloid precursor-like protein 2 （APLP2） C-terminal fragments （CTFs; C57, C50 and C31） in the pEGFP-N1 vector on S100A9 expression in BV2 cells. Reverse transcription-PCR, western blot assay and immunocytochemistry revealed that S100A9 protein and mRNA expression was greater in BV2 cells after CTF transfection than after mock transfection with an empty vector. Furthermore, transfection of full-length APLP2-751 resulted in low levels of S100A9 protein ex-pression. Our results show that APLP2-CTFs upregulate S100A9 protein and mRNA expression in BV2 cells, and identify a novel pathway involved in neuronal injury and apoptosis, and repair and protection in Alzheimer’s disease.
文摘The ability to visualise proteins in their native environment and discern information regarding stoichiometry is of critical importance when studying protein interactions and function. We have used liquid cell atomic force microscopy (AFM) to visualise proteins in their native state in buffer and have determined their molecular volumes. The human proteins S100A8, S100A9, S100A12 and CLIC1 were used in this investigation. The effect of oxidation on the protein structure of CLIC1 was also investigated and we found that CLIC1 multimerisation could be discerned by AFM, which supports similar findings by other methods. We have found good correlation between the molecular volumes measured by AFM and the calculated volumes of the individual proteins. This method allows for the study of single soluble proteins under physiological conditions and could potentially be extended to study the structure of these proteins when located within a membrane environment.
