Objective:To investigate the effects of mechanical stretching and lipopolysaccharide(LPS) on the early apoptosis and IL-8 production of alveolar epithelial type II cells A549.Methods:The experimental matrix consisted ...Objective:To investigate the effects of mechanical stretching and lipopolysaccharide(LPS) on the early apoptosis and IL-8 production of alveolar epithelial type II cells A549.Methods:The experimental matrix consisted of three integrated studies.In the first study,A549 cells were subjected to different stretching strain frequency and duration time to see the effects on the early apoptosis.In the second study,A549 cells were subjected to mechanical stretch(13%4 h, 0.3 Hz) and LPS(1 or 100 ng/mL) to see whether mechanical strain and LPS also have an addictive effect on the early apoptosis.In the third study to investigate whether this addictive effect could be induced by LPS and mechanical stretch on IL-8 production,A549 cells were subjected to LPS(100 ng/mL) and mechanical strain(13%.0.3 Hz,4 h).Real time PCR and enzyme linked immunosorbent assay were used to measure mRN A and protein level of IL-8.The early apoptosis was detected by flow cytometry.Results:Mechanical stretch induced the early apoptosis in a force and frequency and time-dependent manner.In the presence of LPS,mechanical stretch enhanced LPS-induced early apoptosis,especially in 100 ng/mL IPS group compared with 1 ng/ mL LPS and the control group.Mechanical stretch increased IL-8 production and enhanced LPS-induced IL-8 screation both in mRNA and protein levels.Conclusions:Mechanical stretch can induce the early apoptosis and IL-8 secretion.Mechanical stretch and LPS have an addictive effect on the early apoptosis and IL-8 production in alveolar type 2 cells,which is one of the mechanisms of ventilator-induced lung injury.展开更多
TypeⅡdiabetes mellitus(T2DM)has experienced a dramatic increase globally across countries of various income levels over the past three decades.The persistent prevalence of T2DM is attributed to a complex interplay of...TypeⅡdiabetes mellitus(T2DM)has experienced a dramatic increase globally across countries of various income levels over the past three decades.The persistent prevalence of T2DM is attributed to a complex interplay of genetic and environmental factors.While numerous pharmaceutical therapies have been developed,there remains an urgent need for innovative treatment approaches that offer effectiveness without significant adverse effects.In this context,the exploration of the gut microbiome presents a promising avenue.Research has increasingly shown that the gut microbiome of individuals with T2DM exhibits distinct differences compared to healthy individuals,suggesting its potential role in the disease’s pathogenesis and progression.This emerging field offers diverse applications,particularly in modifying the gut environment through the administration of prebiotics,probiotics,and fecal microbiome transfer.These interventions aim to restore a healthy microbiome balance,which could potentially alleviate or even reverse the metabolic dysfunctions associated with T2DM.Although current results from clinical trials have not yet shown dramatic effects on diabetes management,the groundwork has been laid for deeper investigation.Ongoing and future clinical trials are critical to advancing our understanding of the microbiome’s impact on diabetes.By further elucidating the mechanisms through which microbiome alterations influence insulin resistance and glucose metabolism,researchers can develop more targeted interventions.The potential to harness the gut microbiome in developing new therapeutic strategies offers a compelling prospect to transform the treatment landscape of T2DM,potentially reducing the disease’s burden significantly with approaches that are less reliant on traditional pharmaceuticals and more focused on holistic,systemic health improvements.展开更多
Diabetes mellitus is associated with foot ulcers,which frequently pave the way to lower-extremity amputation.Neuropathy,trauma,deformity,high plantar pressures,and peripheral vascular disease are the most common under...Diabetes mellitus is associated with foot ulcers,which frequently pave the way to lower-extremity amputation.Neuropathy,trauma,deformity,high plantar pressures,and peripheral vascular disease are the most common underlying causes.Around 15%of diabetic patients are affected by diabetic foot ulcer in their lifetime.64 million people are affected by diabetics in India and 40000 amputations are done every year.Foot ulcers are evaluated and classified in a systematic and thorough manner to assist in determining the best course of therapy.This paper proposes a novel model which predicts the threat of diabetic foot ulcer using independent agents for various input values and a combination of fuzzy expert systems.The proposed model uses a classification system to distinguish between each fuzzy framework and its parameters.Based on the severity levels necessary prevention,treatment,and medication are recommended.Combining the results of all the fuzzy frameworks derived from its constituent parameters,a risk-specific medication is recommended.The work also has higher accuracy when compared to other related models.展开更多
Based on the observation data of meteorological stations,Doppler radar observation data of Ulanqab City,and ERA-5 reanalysis data,a snowstorm process in Ulanqab City from March 17 to 18,2022 was analyzed.The results s...Based on the observation data of meteorological stations,Doppler radar observation data of Ulanqab City,and ERA-5 reanalysis data,a snowstorm process in Ulanqab City from March 17 to 18,2022 was analyzed.The results show that this was a type Ⅱ snowstorm process generated under the joint influence of upper trough and ground low inverted trough and frontal cyclone.The main period of snowfall can be divided into two time stages,and the total snowfall was more in the south and less in the north,which was consistent with that of average specific humidity field.Water vapor conditions provided by strong water vapor transport and convergence,strong upward movement shown by large vertical velocity field,and the suction action of high-and low-layer divergence and convergence were the reasons for the hourly heavy snowfall on the 18^(th).During the process,radar echoes were mainly sheet-shaped,and composite reflectivity was 15-25 dBZ in most areas.The zero speed line in the first period was positively"S"-shaped,and there was warm advection and southwest wind.On the morning of the 18^(th),after the cold front transited the city,Ulanqab City was gradually controlled by northwest wind,and the snow tended to end.展开更多
针对逐步Ⅱ型删失数据下Burr Type X分布的参数估计问题,提出模型参数的一种新的贝叶斯估计及相应的最大后验密度(HPD)置信区间.假设伽玛分布为待估参数的先验分布,考虑待估参数的条件后验分布未知、单峰且近似对称,选取以正态分布为提...针对逐步Ⅱ型删失数据下Burr Type X分布的参数估计问题,提出模型参数的一种新的贝叶斯估计及相应的最大后验密度(HPD)置信区间.假设伽玛分布为待估参数的先验分布,考虑待估参数的条件后验分布未知、单峰且近似对称,选取以正态分布为提议分布的Metropolis-Hastings(MH)算法生成后验样本,基于后验样本在平方误差损失函数下得到待估参数的贝叶斯估计和HPD置信区间.将基于MH算法得到的贝叶斯估计和HPD置信区间与基于EM算法得到的极大似然估计和置信区间在均方误差准则和精度意义下进行比较.Monte-Carlo模拟结果表明,基于MH算法得到的估计在均方误差准则下优于基于EM算法得到的极大似然估计,基于MH算法得到的HPD置信区间长度小于基于EM算法得到的置信区间长度.展开更多
Summary: The pathogenesis of hyperoxia lung injury and the mechanism of amygdalin on type 2 alveolar epithelial cells (AEC2) isolated from premature rat lungs in vitro were investigated. AEC2 were obtained by primary ...Summary: The pathogenesis of hyperoxia lung injury and the mechanism of amygdalin on type 2 alveolar epithelial cells (AEC2) isolated from premature rat lungs in vitro were investigated. AEC2 were obtained by primary culture from 20-days fetal rat lung and hyperoxia-exposed cell model was established. Cell proliferating viability was examined by MTT assay after treatment of amygdalin at various concentrations. DNA content and the proliferating cell nuclear antigen (PCNA) protein expression of AEC2 were measured by using flow cytometry and immunocytochemistry respectively after 24 h of hyperoxia exposure or amygdalin treatment. The results showed that hyperoxia inhibited the proliferation and decreased PCNA protein expression in AEC2 of premature rat in vitro. Amygdalin at the concentration range of 50-200 μmol/L stimulated the proliferation of AEC2 in a dose-dependent manner, however, 400 μmol/L amygdalin inhibited the proliferation of AEC2. Amygdalin at the concentration of 200 μmol/L played its best role in facilitating proliferation of AEC2s in vitro and could partially ameliorated the changes of proliferation in hyperoxia exposed AEC2 of premature rat. It has been suggested that hyperoxia inhibited the proliferation of AEC2s of premature rat, which may contribute to hyperoxia lung injury. Amygdalin may play partial protective role in hyperoxia-induced lung injury.展开更多
To establish a better method of primary culture for alveolar epithelial type Ⅱ cells （AEC Ⅱ ） and to study its bionomics, alveolar epithelial type Ⅱ cells were isolated by digestion with trypsin and collagenase, ...To establish a better method of primary culture for alveolar epithelial type Ⅱ cells （AEC Ⅱ ） and to study its bionomics, alveolar epithelial type Ⅱ cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin （i The purified AEC Ⅱ were identified by alkaline phosphatase staining, electron mi- croscopy, immunocytochemical staining of pulmonary surfactant protein A （SPA）. The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC Ⅱ could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 × 10^7, and a purity of about 75%-84 %. Cells could be quickly identified with AKP staining. AEC Ⅱ were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC Ⅱ, and AKP staining is simple in cell identification. AEC Ⅱ can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.展开更多
AIM： TO investigate the role of MHC class Ⅱ in the modulation of gastric epithelial cell apoptosis induced by H pylori infection. METHODS： After stimulating a human gastric epithelial cell line with bacteria or ago...AIM： TO investigate the role of MHC class Ⅱ in the modulation of gastric epithelial cell apoptosis induced by H pylori infection. METHODS： After stimulating a human gastric epithelial cell line with bacteria or agonist antibodies specific for MHC class Ⅱ and CD95, the quantitation of apoptotic and anti-apoptotic events, including caspase activation, BCL-2 activation, and FADD recruitment, was performed with a fluorometric assay, a cytometric bead array, and confocal microscopy, respectively. RESULTS： Pretreatment of N87 cells with the anti-MHC class Ⅱ IgM antibody RFD1 resulted in a reduction in global caspase activation at 24 h of H pylori infection. When caspase 3 activation was specifically measured, crosslinking of MHC class Ⅱ resulted in a marked reduced caspase activation, while simple ligation of HHC class Ⅱ did not. Crosslinking of HHC class Ⅱ also resulted in an increased activation of the anti-apoptosis molecule BCL-2 compared to simple ligation. Confocal microscope analysis demonstrated that the pretreatment of gastric epithelial cells with a crosslinking anti-HHC class Ⅱ IgH blocked the recruitment of FADD to the cell surface. CONCLUSION： The results presented here demonstrate that the ability of MHC class Ⅱ to modulate gastric epithelial apoptosis is at least partially dependent on its crosslinking. Furthermore, while previous research has demonstrated that MHC class Ⅱ signaling can be proapoptotic during extended ligation, we have shown that the crosslinking of this molecule has anti-apoptotic ef-fects during the earlier time points of Hpylori infection. This effect is possibly mediated by the ability of MHC class Ⅱ to modulate the activation of the pro-apoptotic receptor Fas by blocking the recruitment of the accessory molecule FADD, and this delay in apoptosis induction could allow for prolonged cytokine secretion by Hpyloriinfected gastric epithelial cells.展开更多
BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients ...BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients were usually affected by heterozygous dominant mutations in the chloride voltage-gated channel 7(ClCN7)gene and present early-onset osteoarthritis or recurrent fractures.In this study,we report a case of persistent joint pain without bone injury or underlying history.CASE SUMMARY We report a 53-year-old female with joint pain who was accidentally diagnosed with ADO-II.The clinical diagnosis was based on increased bone density and typical radiographic features.Two heterozygous mutations in the ClCN7 and Tcell immune regulator 1(TCIRG1)genes by whole exome sequencing were identified in the patient and her daughter.The missense mutation(c.857G>A)occurred in the CLCN7 gene p.R286Q,which is highly conserved across species.The TCIRG1 gene point mutation(c.714-20G>A)in intron 7(near the splicing site of exon 7)had no effect on subsequent transcription.CONCLUSION This ADO-II case had a pathogenic CLCN7 mutation and late onset without the usual clinical symptoms.For the diagnosis and assessment of the prognosis for osteopetrosis,genetic analysis is advised.展开更多
This study investigated the effects of hyperoxia on dynamic changes of thioredoxin-1 （Trx1） and thioredoxin reductase-1 （TrxR1） in alveolar type Ⅱ epithelial cells （AECⅡ） of premature rats. Pregnant Sprague-Da...This study investigated the effects of hyperoxia on dynamic changes of thioredoxin-1 （Trx1） and thioredoxin reductase-1 （TrxR1） in alveolar type Ⅱ epithelial cells （AECⅡ） of premature rats. Pregnant Sprague-Dawley rats were sacrificed on day 19 of gestation. AECⅡ were isolated and purified from the lungs of premature rats. When cultured to 80% confluence, in vitro cells were randomly divided into air group and hyperoxia group. Cells in the hyperoxia group were continuously exposed to 95% O2/5% CO2 and those in the air group to 95% air/5% CO2. After 12, 24 and 48 h, cells in the two groups were harvested to detect their reactive oxygen species （ROS）, apoptosis, TrxR1 activity and the expressions of Trx1 and TrxR1 by corresponding protocols, respectively. The results showed that AECⅡ exposed to hyperoxia generated excessive ROS and the apoptosis percentage in the hyperoxia group was increased significantly at each time points as compared with that in the air group （P0.001）. Moreover, TrxR1 activity was found to be markedly depressed in the hyperoxia group in comparison to that in the air group （P0.001）. RT-PCR showed the expressions of both Trx1 and TrxR1 mRNA were significantly increased in AECⅡ exposed to hyperoxia for 12 and 24 h （P0.01）, respectively. At 48 h, the level of Trx1 mRNA as well as that of TrxR1 mRNA in the hyperoxia group was reduced and showed no significant difference from that in the air group （P0.05）. Western blotting showed the changes of Trx1 protein expressions in the hyperoxia group paralleled those of Trx1 mRNA expressions revealed by RT-PCR. It was concluded that hyperoxia can up-regulate the protective Trx1/TrxR1 expressed by AECⅡ in a certain period, however, also cause dysfunction of the cytoplasmic thioredoxin system by decreasing TrxR1 activity, which may contribute to the progression of oxidative stress and cell apoptosis and finally result in lung injury.展开更多
AIM:To evaluate the efficacy and safety of grasper type scissors(GTS)for endoscopic submucosal dissection(ESD)of gastric epithelial neoplasia.METHODS:The study was performed by 4 endoscopists in 4 institutions affilia...AIM:To evaluate the efficacy and safety of grasper type scissors(GTS)for endoscopic submucosal dissection(ESD)of gastric epithelial neoplasia.METHODS:The study was performed by 4 endoscopists in 4 institutions affiliated to The Catholic University of Korea.ESD was performed in 76 consecutive patients with gastric epithelial neoplasia by using the GTS(37 patients)or the hook knife plus coagrasper(HKC)(39 patients).The complete resection rate,complication rate,total time elapsed and elapsed time per square centimeter of the dissected specimen were analyzed between the GTS and HKC group.RESULTS:The mean age of the GTS group was 62.3±11.4 years and mean age of the HKC group was 65.6±10.1 years.Differentiated adenocarcinoma was found in32.4%in the GTS group and 33.3%in the HKC group.The procedures were performed without interruption in every case in both groups.The en bloc resection rates of both groups were 100%.The total time elapsed during the procedure was 44.54±21.72 min in the GTS group and 43.77±21.84 min in the HKC group(P=0.88)and the time elapsed per square centimeter of the resected lesion was 7.53±6.35 min/cm2in the GTS group and 6.92±5.93 min/cm2in the HKC group(P=0.66).The overall complication rate was not significantly different between the two groups.CONCLUSION:GTS is a safe and effective device for ESD compared with HKC.ESD can be performed with GTS alone,which can reduce the costs for ESD.展开更多
Objective： To explore the functions and mechanisms of herpes simplex virus type I（HSV-1） while infecting human oral epithelial cells in vitro（being similar to the infection in vivo）. Methods：An abundance of HSV-...Objective： To explore the functions and mechanisms of herpes simplex virus type I（HSV-1） while infecting human oral epithelial cells in vitro（being similar to the infection in vivo）. Methods：An abundance of HSV-1 strains amplified in Vero cells were used to infect human oral epithelial cells. The culture supernatant was collected to infect Vero cells again. Morphology of HSV-1 was identified by inverted microscope and transmission electron microscope. Nucleic acid of the virus was detected by PCR. Results：The infected human oral epithelial cells didn＇ t display an obvious cytopathic effect（CPE） under inverted microscope（while Vero cells which were infected by the culture supernatant showed typical（CPE）. The virus particles were not observed in the cytoplasm nor in nucleus of human oral epithelial cells, however under transmission electron microscope in the cytoplasm of Vero cells, the nucleic acid of HSV-1 could be detected in infected human oral epithelial cells, by PCR. Conclusion-HSV-1 can successfully infect human oral epithelial cells. This model may provide a useful approach for studying the pathogenesis of herpes virus-associated periodontal disease.展开更多
Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubula...Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.展开更多
文摘Objective:To investigate the effects of mechanical stretching and lipopolysaccharide(LPS) on the early apoptosis and IL-8 production of alveolar epithelial type II cells A549.Methods:The experimental matrix consisted of three integrated studies.In the first study,A549 cells were subjected to different stretching strain frequency and duration time to see the effects on the early apoptosis.In the second study,A549 cells were subjected to mechanical stretch(13%4 h, 0.3 Hz) and LPS(1 or 100 ng/mL) to see whether mechanical strain and LPS also have an addictive effect on the early apoptosis.In the third study to investigate whether this addictive effect could be induced by LPS and mechanical stretch on IL-8 production,A549 cells were subjected to LPS(100 ng/mL) and mechanical strain(13%.0.3 Hz,4 h).Real time PCR and enzyme linked immunosorbent assay were used to measure mRN A and protein level of IL-8.The early apoptosis was detected by flow cytometry.Results:Mechanical stretch induced the early apoptosis in a force and frequency and time-dependent manner.In the presence of LPS,mechanical stretch enhanced LPS-induced early apoptosis,especially in 100 ng/mL IPS group compared with 1 ng/ mL LPS and the control group.Mechanical stretch increased IL-8 production and enhanced LPS-induced IL-8 screation both in mRNA and protein levels.Conclusions:Mechanical stretch can induce the early apoptosis and IL-8 secretion.Mechanical stretch and LPS have an addictive effect on the early apoptosis and IL-8 production in alveolar type 2 cells,which is one of the mechanisms of ventilator-induced lung injury.
文摘TypeⅡdiabetes mellitus(T2DM)has experienced a dramatic increase globally across countries of various income levels over the past three decades.The persistent prevalence of T2DM is attributed to a complex interplay of genetic and environmental factors.While numerous pharmaceutical therapies have been developed,there remains an urgent need for innovative treatment approaches that offer effectiveness without significant adverse effects.In this context,the exploration of the gut microbiome presents a promising avenue.Research has increasingly shown that the gut microbiome of individuals with T2DM exhibits distinct differences compared to healthy individuals,suggesting its potential role in the disease’s pathogenesis and progression.This emerging field offers diverse applications,particularly in modifying the gut environment through the administration of prebiotics,probiotics,and fecal microbiome transfer.These interventions aim to restore a healthy microbiome balance,which could potentially alleviate or even reverse the metabolic dysfunctions associated with T2DM.Although current results from clinical trials have not yet shown dramatic effects on diabetes management,the groundwork has been laid for deeper investigation.Ongoing and future clinical trials are critical to advancing our understanding of the microbiome’s impact on diabetes.By further elucidating the mechanisms through which microbiome alterations influence insulin resistance and glucose metabolism,researchers can develop more targeted interventions.The potential to harness the gut microbiome in developing new therapeutic strategies offers a compelling prospect to transform the treatment landscape of T2DM,potentially reducing the disease’s burden significantly with approaches that are less reliant on traditional pharmaceuticals and more focused on holistic,systemic health improvements.
文摘Diabetes mellitus is associated with foot ulcers,which frequently pave the way to lower-extremity amputation.Neuropathy,trauma,deformity,high plantar pressures,and peripheral vascular disease are the most common underlying causes.Around 15%of diabetic patients are affected by diabetic foot ulcer in their lifetime.64 million people are affected by diabetics in India and 40000 amputations are done every year.Foot ulcers are evaluated and classified in a systematic and thorough manner to assist in determining the best course of therapy.This paper proposes a novel model which predicts the threat of diabetic foot ulcer using independent agents for various input values and a combination of fuzzy expert systems.The proposed model uses a classification system to distinguish between each fuzzy framework and its parameters.Based on the severity levels necessary prevention,treatment,and medication are recommended.Combining the results of all the fuzzy frameworks derived from its constituent parameters,a risk-specific medication is recommended.The work also has higher accuracy when compared to other related models.
文摘Based on the observation data of meteorological stations,Doppler radar observation data of Ulanqab City,and ERA-5 reanalysis data,a snowstorm process in Ulanqab City from March 17 to 18,2022 was analyzed.The results show that this was a type Ⅱ snowstorm process generated under the joint influence of upper trough and ground low inverted trough and frontal cyclone.The main period of snowfall can be divided into two time stages,and the total snowfall was more in the south and less in the north,which was consistent with that of average specific humidity field.Water vapor conditions provided by strong water vapor transport and convergence,strong upward movement shown by large vertical velocity field,and the suction action of high-and low-layer divergence and convergence were the reasons for the hourly heavy snowfall on the 18^(th).During the process,radar echoes were mainly sheet-shaped,and composite reflectivity was 15-25 dBZ in most areas.The zero speed line in the first period was positively"S"-shaped,and there was warm advection and southwest wind.On the morning of the 18^(th),after the cold front transited the city,Ulanqab City was gradually controlled by northwest wind,and the snow tended to end.
文摘针对逐步Ⅱ型删失数据下Burr Type X分布的参数估计问题,提出模型参数的一种新的贝叶斯估计及相应的最大后验密度(HPD)置信区间.假设伽玛分布为待估参数的先验分布,考虑待估参数的条件后验分布未知、单峰且近似对称,选取以正态分布为提议分布的Metropolis-Hastings(MH)算法生成后验样本,基于后验样本在平方误差损失函数下得到待估参数的贝叶斯估计和HPD置信区间.将基于MH算法得到的贝叶斯估计和HPD置信区间与基于EM算法得到的极大似然估计和置信区间在均方误差准则和精度意义下进行比较.Monte-Carlo模拟结果表明,基于MH算法得到的估计在均方误差准则下优于基于EM算法得到的极大似然估计,基于MH算法得到的HPD置信区间长度小于基于EM算法得到的置信区间长度.
文摘Summary: The pathogenesis of hyperoxia lung injury and the mechanism of amygdalin on type 2 alveolar epithelial cells (AEC2) isolated from premature rat lungs in vitro were investigated. AEC2 were obtained by primary culture from 20-days fetal rat lung and hyperoxia-exposed cell model was established. Cell proliferating viability was examined by MTT assay after treatment of amygdalin at various concentrations. DNA content and the proliferating cell nuclear antigen (PCNA) protein expression of AEC2 were measured by using flow cytometry and immunocytochemistry respectively after 24 h of hyperoxia exposure or amygdalin treatment. The results showed that hyperoxia inhibited the proliferation and decreased PCNA protein expression in AEC2 of premature rat in vitro. Amygdalin at the concentration range of 50-200 μmol/L stimulated the proliferation of AEC2 in a dose-dependent manner, however, 400 μmol/L amygdalin inhibited the proliferation of AEC2. Amygdalin at the concentration of 200 μmol/L played its best role in facilitating proliferation of AEC2s in vitro and could partially ameliorated the changes of proliferation in hyperoxia exposed AEC2 of premature rat. It has been suggested that hyperoxia inhibited the proliferation of AEC2s of premature rat, which may contribute to hyperoxia lung injury. Amygdalin may play partial protective role in hyperoxia-induced lung injury.
基金the National Natural Sciences Foundation of China (No. 30500224 and No. 30400193).
文摘To establish a better method of primary culture for alveolar epithelial type Ⅱ cells （AEC Ⅱ ） and to study its bionomics, alveolar epithelial type Ⅱ cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin （i The purified AEC Ⅱ were identified by alkaline phosphatase staining, electron mi- croscopy, immunocytochemical staining of pulmonary surfactant protein A （SPA）. The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC Ⅱ could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 × 10^7, and a purity of about 75%-84 %. Cells could be quickly identified with AKP staining. AEC Ⅱ were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC Ⅱ, and AKP staining is simple in cell identification. AEC Ⅱ can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
基金Supported by the National Institutes of Health Grants DK50669, DK56338 and National Institutes of Health T32 AI007536-06 Training Grant
文摘AIM： TO investigate the role of MHC class Ⅱ in the modulation of gastric epithelial cell apoptosis induced by H pylori infection. METHODS： After stimulating a human gastric epithelial cell line with bacteria or agonist antibodies specific for MHC class Ⅱ and CD95, the quantitation of apoptotic and anti-apoptotic events, including caspase activation, BCL-2 activation, and FADD recruitment, was performed with a fluorometric assay, a cytometric bead array, and confocal microscopy, respectively. RESULTS： Pretreatment of N87 cells with the anti-MHC class Ⅱ IgM antibody RFD1 resulted in a reduction in global caspase activation at 24 h of H pylori infection. When caspase 3 activation was specifically measured, crosslinking of MHC class Ⅱ resulted in a marked reduced caspase activation, while simple ligation of HHC class Ⅱ did not. Crosslinking of HHC class Ⅱ also resulted in an increased activation of the anti-apoptosis molecule BCL-2 compared to simple ligation. Confocal microscope analysis demonstrated that the pretreatment of gastric epithelial cells with a crosslinking anti-HHC class Ⅱ IgH blocked the recruitment of FADD to the cell surface. CONCLUSION： The results presented here demonstrate that the ability of MHC class Ⅱ to modulate gastric epithelial apoptosis is at least partially dependent on its crosslinking. Furthermore, while previous research has demonstrated that MHC class Ⅱ signaling can be proapoptotic during extended ligation, we have shown that the crosslinking of this molecule has anti-apoptotic ef-fects during the earlier time points of Hpylori infection. This effect is possibly mediated by the ability of MHC class Ⅱ to modulate the activation of the pro-apoptotic receptor Fas by blocking the recruitment of the accessory molecule FADD, and this delay in apoptosis induction could allow for prolonged cytokine secretion by Hpyloriinfected gastric epithelial cells.
基金Supported by the Science and Technology Plan Program of Sichuan of China,No.2018JY0608。
文摘BACKGROUND Osteopetrosis is a rare genetic disorder characterized by increased bone density due to defective bone resorption of osteoclasts.Approximately,80%of autosomal dominant osteopetrosis type II(ADO-II)patients were usually affected by heterozygous dominant mutations in the chloride voltage-gated channel 7(ClCN7)gene and present early-onset osteoarthritis or recurrent fractures.In this study,we report a case of persistent joint pain without bone injury or underlying history.CASE SUMMARY We report a 53-year-old female with joint pain who was accidentally diagnosed with ADO-II.The clinical diagnosis was based on increased bone density and typical radiographic features.Two heterozygous mutations in the ClCN7 and Tcell immune regulator 1(TCIRG1)genes by whole exome sequencing were identified in the patient and her daughter.The missense mutation(c.857G>A)occurred in the CLCN7 gene p.R286Q,which is highly conserved across species.The TCIRG1 gene point mutation(c.714-20G>A)in intron 7(near the splicing site of exon 7)had no effect on subsequent transcription.CONCLUSION This ADO-II case had a pathogenic CLCN7 mutation and late onset without the usual clinical symptoms.For the diagnosis and assessment of the prognosis for osteopetrosis,genetic analysis is advised.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30770944)
文摘This study investigated the effects of hyperoxia on dynamic changes of thioredoxin-1 （Trx1） and thioredoxin reductase-1 （TrxR1） in alveolar type Ⅱ epithelial cells （AECⅡ） of premature rats. Pregnant Sprague-Dawley rats were sacrificed on day 19 of gestation. AECⅡ were isolated and purified from the lungs of premature rats. When cultured to 80% confluence, in vitro cells were randomly divided into air group and hyperoxia group. Cells in the hyperoxia group were continuously exposed to 95% O2/5% CO2 and those in the air group to 95% air/5% CO2. After 12, 24 and 48 h, cells in the two groups were harvested to detect their reactive oxygen species （ROS）, apoptosis, TrxR1 activity and the expressions of Trx1 and TrxR1 by corresponding protocols, respectively. The results showed that AECⅡ exposed to hyperoxia generated excessive ROS and the apoptosis percentage in the hyperoxia group was increased significantly at each time points as compared with that in the air group （P0.001）. Moreover, TrxR1 activity was found to be markedly depressed in the hyperoxia group in comparison to that in the air group （P0.001）. RT-PCR showed the expressions of both Trx1 and TrxR1 mRNA were significantly increased in AECⅡ exposed to hyperoxia for 12 and 24 h （P0.01）, respectively. At 48 h, the level of Trx1 mRNA as well as that of TrxR1 mRNA in the hyperoxia group was reduced and showed no significant difference from that in the air group （P0.05）. Western blotting showed the changes of Trx1 protein expressions in the hyperoxia group paralleled those of Trx1 mRNA expressions revealed by RT-PCR. It was concluded that hyperoxia can up-regulate the protective Trx1/TrxR1 expressed by AECⅡ in a certain period, however, also cause dysfunction of the cytoplasmic thioredoxin system by decreasing TrxR1 activity, which may contribute to the progression of oxidative stress and cell apoptosis and finally result in lung injury.
基金Supported by Research grants from Korean Society of Gastrointestinal Endoscopy
文摘AIM:To evaluate the efficacy and safety of grasper type scissors(GTS)for endoscopic submucosal dissection(ESD)of gastric epithelial neoplasia.METHODS:The study was performed by 4 endoscopists in 4 institutions affiliated to The Catholic University of Korea.ESD was performed in 76 consecutive patients with gastric epithelial neoplasia by using the GTS(37 patients)or the hook knife plus coagrasper(HKC)(39 patients).The complete resection rate,complication rate,total time elapsed and elapsed time per square centimeter of the dissected specimen were analyzed between the GTS and HKC group.RESULTS:The mean age of the GTS group was 62.3±11.4 years and mean age of the HKC group was 65.6±10.1 years.Differentiated adenocarcinoma was found in32.4%in the GTS group and 33.3%in the HKC group.The procedures were performed without interruption in every case in both groups.The en bloc resection rates of both groups were 100%.The total time elapsed during the procedure was 44.54±21.72 min in the GTS group and 43.77±21.84 min in the HKC group(P=0.88)and the time elapsed per square centimeter of the resected lesion was 7.53±6.35 min/cm2in the GTS group and 6.92±5.93 min/cm2in the HKC group(P=0.66).The overall complication rate was not significantly different between the two groups.CONCLUSION:GTS is a safe and effective device for ESD compared with HKC.ESD can be performed with GTS alone,which can reduce the costs for ESD.
文摘Objective： To explore the functions and mechanisms of herpes simplex virus type I（HSV-1） while infecting human oral epithelial cells in vitro（being similar to the infection in vivo）. Methods：An abundance of HSV-1 strains amplified in Vero cells were used to infect human oral epithelial cells. The culture supernatant was collected to infect Vero cells again. Morphology of HSV-1 was identified by inverted microscope and transmission electron microscope. Nucleic acid of the virus was detected by PCR. Results：The infected human oral epithelial cells didn＇ t display an obvious cytopathic effect（CPE） under inverted microscope（while Vero cells which were infected by the culture supernatant showed typical（CPE）. The virus particles were not observed in the cytoplasm nor in nucleus of human oral epithelial cells, however under transmission electron microscope in the cytoplasm of Vero cells, the nucleic acid of HSV-1 could be detected in infected human oral epithelial cells, by PCR. Conclusion-HSV-1 can successfully infect human oral epithelial cells. This model may provide a useful approach for studying the pathogenesis of herpes virus-associated periodontal disease.
文摘Objective Ubiquitin-specific protease 4(USP4)facilitates the development of transforming growth factor-beta 1(TGF-β1)-induced epithelial-mesenchymal transition(EMT)in various cancer cells.Moreover,EMT of renal tubular epithelial cells(RTECs)is required for the progression of renal interstitial fibrosis.However,the role of USP4 in EMT of RTECs remains unknown.The present study aimed to explore the effect of USP4 on the EMT of RTECs as well as the involved mechanism.Methods In established unilateral ureteral obstruction(UUO)rats and NRK-52E cells,immunohistochemistry and Western blot assays were performed.Results USP4 expression was increased significantly with obstruction time.In NRK-52E cells stimulated by TGF-β1,USP4 expression was increased in a time-dependent manner.In addition,USP4 silencing with specific siRNA indicated that USP4 protein was suppressed effectively.Meanwhile,USP4 siRNA treatment restored E-cadherin and weakened alpha smooth muscle actin(α-SMA)expression,indicating that USP4 may promote EMT.After treatment with USP4 siRNA and TGF-β1 for 24 h,the expression of TGF-β1 receptor type I(TβRI)was decreased.Conclusion USP4 promotes the EMT of RTECs through upregulating TβRI,thereby facilitating renal interstitial fibrosis.These findings may provide a potential target of USP4 in the treatment of renal fibrosis.
