Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephad...Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephadex G-15 gel chromatography and RP-HPLC. One novel anti-hepatic fibrosis peptide （CTEPH-I： Asn-Pro-Asn-Pro-Thr） was obtained and identified. MTS assay and enzyme-linked immunosorbent assay were applied to evaluate the anti-fibrotic effect of CTEPH-1 on activated hepatic stellate cells （HSCs） in vitro. CTEPH-1 efficiently inhibited activation and proliferation of cultured HSC-T6 cells via lowering the contents of collagen and TIMP- 1 except for matrix metalloproteinase- 1 （MMP- 1）. The purified peptide might be beneficial as functional food or potential drug for treatment of liver fibrogenesis.展开更多
Heating,Ventilation,andAir Conditioning(HVAC)systems are critical formaintaining thermal comfort in office environments which also crucial for occupant well-being and productivity.This study investigates the impact of...Heating,Ventilation,andAir Conditioning(HVAC)systems are critical formaintaining thermal comfort in office environments which also crucial for occupant well-being and productivity.This study investigates the impact of integrating ceiling fans with higher air conditioning setpoints on thermal comfort and energy efficiency in office environments.Field measurements and questionnaire surveys were conducted to evaluate thermal comfort and energysaving potential under varying conditions.Results show that increasing the AC setpoint from 25○C to 27○C,combined with ceiling fan operation,reduced power consumption by 10%,achieving significant energy savings.Survey data confirmed that 85%of participants reported consistent thermal sensations across all conditions,with ceiling fans effectively compensating for higher setpoints through enhanced air circulation.CFDsimulations revealed that mediumspeed ceiling fan operation produced the most uniformairflowdistribution,with an average air velocity of 0.45 m/s,and minimized temperature variations,ensuring balanced thermal conditions.Temperature analysis showed a reduction in hotspots and cold zones,maintaining an average temperature deviation of less than±0.5○C.Predicted Mean Vote(PMV)evaluations at a 27○C setpoint indicated improved thermal comfort,with average PMV values around−0.3,corresponding to a“neutral”thermal sensation.These findings demonstrate the effectiveness of integrating ceiling fans with HVAC systems in achieving energy efficiency and occupant comfort,offering a sustainable approach to reducing AC energy consumption in office environments.展开更多
Sericin from discarded silkworm cocoons of silk reeling has been used in different fields, such as cosmetology, skin care, nutrition, and oncology. The present study established a rat model of type 2 diabetes by conse...Sericin from discarded silkworm cocoons of silk reeling has been used in different fields, such as cosmetology, skin care, nutrition, and oncology. The present study established a rat model of type 2 diabetes by consecutive intraperitoneal injections of low-dose （25 mg/kg） streptozotocin. After intragastrical perfusion of sericin for 35 days, blood glucose levels significantly declined, and the expression of neurofilament protein in the sciatic nerve and nerve growth factor in L4-6 spinal ganglion and anterior horn cells significantly increased. However, the expression of neuropeptide Y in spinal ganglion and anterior horn cells significantly decreased in model rats. These findings indicate that sericin protected the sciatic nerve and related nerve cells against injury in a rat type 2 diabetic model by upregulating the expression of neurofilament protein in the sciatic nerve and nerve growth factor in spinal ganglion and anterior horn cells, and downregulating the expression of neuropeptide Y in spinal ganglion and anterior horn cells.展开更多
A non-noble metal oxygen reduction reaction （ORR） catalyst labeled as Co-C-N（800） was synthesized by heat-treating a mixture of urea, cobalt chloride and acetylene black for 2 h at 800 ℃ in an inert nitrogen atmo...A non-noble metal oxygen reduction reaction （ORR） catalyst labeled as Co-C-N（800） was synthesized by heat-treating a mixture of urea, cobalt chloride and acetylene black for 2 h at 800 ℃ in an inert nitrogen atmosphere. X-ray diffraction pattern indicates that a metallic β-Co is generated after the heat-treating process. The results from cyclic voltammograms show that the obtained Co-C-N（800） catalyst has good ORR catalytic activity in 0.5 mol/L H2SO4 solution. The catalyst is also good at methanol tolerance and stability in the acidic solution.展开更多
Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small...Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.展开更多
Summary： Previous studies have shown that STAT3 plays a vital role in the genesis and progression of cancer. In this study, we investigated the relationship between the JAK2/STAT3 signalling pathway and germacrone-in...Summary： Previous studies have shown that STAT3 plays a vital role in the genesis and progression of cancer. In this study, we investigated the relationship between the JAK2/STAT3 signalling pathway and germacrone-induced apoptosis in HepG2 cells. HepG2 cells were incubated with germacrone for 24 h, the protein expression of p-STAT3, STAT3, p-JAK2 and JAK2 was detected by Westem Blotting, and RT-PCR was used to determine the expression of STAT3, p53, Bcl-2 and Bax at transcriptional levels. Besides that, HepG2 cells were pre-treated with AG490 or IL-6 for 2 h, and then incubated with ger- macrone for 24 h. The expression ofp-JAK2, JAK2, p-STAT3, STAT3, p53, Bax and Bcl-2 was detected by Western blotting. The activity of HepG2 cells was tested by MTT assay. The apoptosis of HepG2 cells and levels of reactive oxygen species （ROS） were flow cytometrically measured. The results showed that germacrone exposure decreased p-STAT3 and p-JAK2 and regulated expression of p53 and Bcl-2 family members at the same time. Moreover, IL-6 enhanced the activation of the JAK2/STAT3 signalling pathway and therefore attenuated the germacrone-induced apoptosis. Suppression of JAK2/STAT3 signalling pathway by AG490, an inhibitor of JAK2, resulted in apoptosis and an increase in ROS in response to germacrone exposure. We therefore conclude that germacrone induces apoptosis through the JAK2/STAT3 signalling pathway.展开更多
Water pollution of the Yangtze River in China became one of challenges that the government is facing today. Increasing numbers of petrochemical plants were built along the river in past decades, and numbers of organic...Water pollution of the Yangtze River in China became one of challenges that the government is facing today. Increasing numbers of petrochemical plants were built along the river in past decades, and numbers of organic chemicals were discharged into the river. Our goal was to establish in vitro system on rat sertoli cells, spermatogenic cells and leydig cells to investigate the reproductive toxicity potential induced by organic extracts from petrochemical effluents. Our results showed that the organic extract depressed the viability （p 〈 0.01） and destroyed the plasma membrane integrity of sertoli cells and spermatogenic cells to a certain degree. Accordingly, proportion of early apoptotic sertoli cells and late apoptotic spermatogenic cells increased significantly. Although significant morphological changes were not detected for leydig cells, the extract was observed to inhibit their testosterone production （p 〈 0.01）. Sertoli cells and sperrnatogenic cells appeared to be more sensitive and maybe the main targets of the key toxins. These results suggested that the in vitro system on rat testicular cells may be useful to predicate reproductive toxicity potential of organic extracts from petrochemical effluents. More attention should be paid to the petrochemical effluents, because long-term accumulation of these organic compounds in organisms may cause spermatogenesis malfunction and testosterone reduction.展开更多
Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） subst...Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） substrate in vitro. There are indications that they may simultaneously lose their sternness and multipotentiality as the result of prolonged 2D culture. In this study, we used three-dimensional （3D） collagen scaffolds as rat MSCs cartier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs. The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation. More importantly, compared to MSCs under 2D culture, 3D MSCs significantly maintained higher expression levels of stemness genes （Oct4, Sox2, Rex-1 and Nanog）, yielded high frequencies of colony-forming units-fibroblastic （CFU-F） and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction. Thus, 3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.展开更多
The expression and implication of vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （MMP-9） in residual hepatic tumor cells after lipiodol embolization were investi- gated. Two weeks after t...The expression and implication of vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （MMP-9） in residual hepatic tumor cells after lipiodol embolization were investi- gated. Two weeks after transplantation of VX2 tumor cells into the livers of rabbits, a xenograft model of the human hepatic neoplasm was successfully established. Forty rabbits were randomly divided into control group （n=20） and lipiodol group （n=20）. For the control group, 1 mL normal saline was injected through the gastroduodenal artery, whereas 0.3 mL/kg lipiodol was applied for the lipiodol group. One week after embolization, the expression level of VEGF in the plasma was measured by using en- zyme-linked immunosorbent assay （ELISA）. A three-step immunohistochemieal technique （ABC） was employed to detect the protein levels of VEGF and MMP-9 and the quantitative PCR for their mRNA levels was performed in the residual tumor cells. The VEGF in the plasma was significantly higher in the lipiodol group （1.42~0.29 ng/mL） than in the control group （1.12~0.21 ng/mL） （P〈0.01）. Moreover, the positive rate of VEGF protein in the residual tumor cells was significantly higher in the lipiodol group （62.13%~7.69%） than in the control group （53.16%~9.17%） （P〈0.05）. Similarly, the MMP-9 ex- pression in the residual ~mor cells was higher in the lipiodol group. The mRNA levels of VEGF （2.9313~2.4231） and MMP-9 （3.5721~1.6107） in the lipiodol group were significantly higher than those in the control group （1.5728~0.9453 and 1.7573~1.0641, respectively, P〈0.05）. Therefore, it was rea- sonable to speculate that the increased expression of VEGF and MMP-9 in residual hepatic tumor cells and tumor angiogenesis post-embolization would be responsible for the increased metastatic potentiality and invasiveness of these cells.展开更多
Circulating tumor cells （CTCs） arise from primary or secondary tumors and enter the bloodstream by active or passive intravasation. Given the low number of CTCs, enrichment is necessary for detection. Filtration met...Circulating tumor cells （CTCs） arise from primary or secondary tumors and enter the bloodstream by active or passive intravasation. Given the low number of CTCs, enrichment is necessary for detection. Filtration methods are based on selection of CTCs by size using a filter with 6.5 to 8 pm pores. After coloration, collected CTCs are evaluated according to morphological criteria. Immunophenotyping and fluorescence in situ hybridization techniques may be used. Selected CTCs can also be cultivated in vitro to provide more material. Analysis of genomic mutations is difficult because it requires adapted techniques due to limited DNA materials. Filtration-selected CTCs have shown prognostic value in many studies but multicentric validating trials are mandatory to strengthen this assessment. Other clinical applications are promising such as follow-up, therapy response prediction and diagnosis. Microfluidic emerging systems could optimize filtration-selected CTCs by increasing selection accuracy.展开更多
Embryonic germ （EG） cells are cultured pluripotent stem cells derived from the primordial germ cells （PGCs） that migrate from the dorsal mesentery of the hindgut to the developing genital ridge. In this study, the...Embryonic germ （EG） cells are cultured pluripotent stem cells derived from the primordial germ cells （PGCs） that migrate from the dorsal mesentery of the hindgut to the developing genital ridge. In this study, the morphology of the porcine genital ridge was assessed in embryos harvested on days 22-30 of pregnancy. PGCs from embryos at these stages were cultured to obtain porcine EG cell lines, and EG-like cells were derived from PGCs from embryos harvested on days 24-28 of pregnancy. The EG-like cells expressed Oct4, Sox2, Nanog, SSEA-3, SSEA-4 and alkaline phosphatase （AP）. These cells were able to form embryoid bodies （EBs） in suspension culture and differentiate into cells representative of the three germ layers as verified by a-fetoprotein （AFP）, s-smooth muscle actin （^-SMA）, and Nestin expression. Spontaneous differentiation from the porcine EG-like cells of delayed passage in vitro showed that they could differentiate into epithelial-like cells, mesenchymal-like cells and neuron-like cells. In vitro directed differentiation generated osteocytes, adipocytes and a variety of neural lineage cells, as demonstrated by alizarin red staining, oil red O staining, and immunoftuorescence for neuronal class III [3-tubulin （Tuj 1）, glial fibrillary protein （GFAP） and galactosylceramidase （GALC）, respectively. These results indicate that porcine EG-like cells have the potential for multi-lineage differentiation and are useful for basic porcine stem cell research.展开更多
Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers ...Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.展开更多
Estrogen-related receptor alpha （ERRα） plays an important role in the development of hor- monezdependent cancers, but its roles in lung cancer remain elusive. The present study was aimed to investigate the effects ...Estrogen-related receptor alpha （ERRα） plays an important role in the development of hor- monezdependent cancers, but its roles in lung cancer remain elusive. The present study was aimed to investigate the effects of ERRα on the proliferation and metastasis of lung cancer A549 cells. The mRNA and protein levels of ERRor were detected in lung cancer A549 and MCF-7 cells and bronchial epithelial BEAS-2B cells by qRT-PCR and Western blotting, respectively. ERRor plasmid transfection and XCT-790 （an inverse agonist of ERRc0 were used to up-regulate or down-regulate ERRα expression in A549 cells, respectively. The viability of A549 cells was measured by cell counting kit-8 （CCK-8） and the motility of A549 cells by wound healing assay and Transwell migration/invasion assay. The epithelial markers E-cadherin （E-Cad） and zona occludin-1 （ZO-1）, the mesenchymal markers fi- bronectin （FN） and vimentin （Vim） and the transcription factors （Snail, Zebl Twist and Slug） were fur- ther detected at mRNA and protein levels by qRT-PCR and Western blotting, respectively. The results showed that ERRor promoted the growth of lung cancer A549 cells in vitro. XCT-790 significantly in- hibited the migration and invasion of A549 cells. Over-expression of ERRα promoted the epithe- lial-to-mesenchymal transition （EMT） of A549 ceils, down-regulated the epithelial makers E-Cad and ZO-1, and up-regulated the mesenchymal makers FN and Vim. Silencing of Slug, but not other tran- scription factors, significantly abolished the ERRs-induced EMT of A549 cells. It was suggested that ERRor promoted the migration and invasion of A549 cells by inducing EMT, and Slug was involved in the process. Targeting ERRor might be an efficient approach for lung cancer treatment.展开更多
Cultured Schwann cells were treated with 5.6 mM and 50 mM glucose alternating every 8 hours to simulate intermittent high glucose. The present study analyzed the neuroprotective effects of 1, 10 and 100 μM ginsenosid...Cultured Schwann cells were treated with 5.6 mM and 50 mM glucose alternating every 8 hours to simulate intermittent high glucose. The present study analyzed the neuroprotective effects of 1, 10 and 100 μM ginsenoside Rbl on oxidative damage and apoptosis in Schwann cells induced by intermittent high glucose. Flow cytometry demonstrated that ginsenoside Rbl reduced intermittent high glucose-mediated reactive oxygen species production. Enzyme linked immunosorbent assay showed that 8-hydroxy-2-deoxy guanosine levels in Schwann cells decreased following ginsenoside Rbl treatment. Quantitative real-time reverse transcription-PCR and western blot assay results revealed that ginsenoside Rbl inhibited intermittent high glucose-upregulated Bax expression, but antagonized intermittent high glucose-downregulated Bcl-2 expression in Schwann cells. These effects were most pronounced with 100 μM ginsenoside Rbl. These results indicate that ginsenoside Rbl inhibits intermittent high glucose-induced oxidative stress and apoptosis in Schwann cells.展开更多
基金The Foundation of the Education Department of Hubei Province(Grant No.D20162004)
文摘Anti-hepatic fibrosis peptide from Carapax Trionycis was purified, characterized, and inhibitory effect was assessed. Carapax Trionycis extract peptide hydrolysates （CTEPHs） were separated by ultrafiltration, Sephadex G-15 gel chromatography and RP-HPLC. One novel anti-hepatic fibrosis peptide （CTEPH-I： Asn-Pro-Asn-Pro-Thr） was obtained and identified. MTS assay and enzyme-linked immunosorbent assay were applied to evaluate the anti-fibrotic effect of CTEPH-1 on activated hepatic stellate cells （HSCs） in vitro. CTEPH-1 efficiently inhibited activation and proliferation of cultured HSC-T6 cells via lowering the contents of collagen and TIMP- 1 except for matrix metalloproteinase- 1 （MMP- 1）. The purified peptide might be beneficial as functional food or potential drug for treatment of liver fibrogenesis.
基金support by the National Science and Technology Council under Grant No.NSTC 112-2221-E-167-017-MY3.
文摘Heating,Ventilation,andAir Conditioning(HVAC)systems are critical formaintaining thermal comfort in office environments which also crucial for occupant well-being and productivity.This study investigates the impact of integrating ceiling fans with higher air conditioning setpoints on thermal comfort and energy efficiency in office environments.Field measurements and questionnaire surveys were conducted to evaluate thermal comfort and energysaving potential under varying conditions.Results show that increasing the AC setpoint from 25○C to 27○C,combined with ceiling fan operation,reduced power consumption by 10%,achieving significant energy savings.Survey data confirmed that 85%of participants reported consistent thermal sensations across all conditions,with ceiling fans effectively compensating for higher setpoints through enhanced air circulation.CFDsimulations revealed that mediumspeed ceiling fan operation produced the most uniformairflowdistribution,with an average air velocity of 0.45 m/s,and minimized temperature variations,ensuring balanced thermal conditions.Temperature analysis showed a reduction in hotspots and cold zones,maintaining an average temperature deviation of less than±0.5○C.Predicted Mean Vote(PMV)evaluations at a 27○C setpoint indicated improved thermal comfort,with average PMV values around−0.3,corresponding to a“neutral”thermal sensation.These findings demonstrate the effectiveness of integrating ceiling fans with HVAC systems in achieving energy efficiency and occupant comfort,offering a sustainable approach to reducing AC energy consumption in office environments.
文摘Sericin from discarded silkworm cocoons of silk reeling has been used in different fields, such as cosmetology, skin care, nutrition, and oncology. The present study established a rat model of type 2 diabetes by consecutive intraperitoneal injections of low-dose （25 mg/kg） streptozotocin. After intragastrical perfusion of sericin for 35 days, blood glucose levels significantly declined, and the expression of neurofilament protein in the sciatic nerve and nerve growth factor in L4-6 spinal ganglion and anterior horn cells significantly increased. However, the expression of neuropeptide Y in spinal ganglion and anterior horn cells significantly decreased in model rats. These findings indicate that sericin protected the sciatic nerve and related nerve cells against injury in a rat type 2 diabetic model by upregulating the expression of neurofilament protein in the sciatic nerve and nerve growth factor in spinal ganglion and anterior horn cells, and downregulating the expression of neuropeptide Y in spinal ganglion and anterior horn cells.
文摘A non-noble metal oxygen reduction reaction （ORR） catalyst labeled as Co-C-N（800） was synthesized by heat-treating a mixture of urea, cobalt chloride and acetylene black for 2 h at 800 ℃ in an inert nitrogen atmosphere. X-ray diffraction pattern indicates that a metallic β-Co is generated after the heat-treating process. The results from cyclic voltammograms show that the obtained Co-C-N（800） catalyst has good ORR catalytic activity in 0.5 mol/L H2SO4 solution. The catalyst is also good at methanol tolerance and stability in the acidic solution.
基金supported by grants from Wuhan Municipal Science and Technology Research Project,China(No.201260523185)the Public Science and Technology Research Funds Projects of Ocean,China(No.201005013)
文摘Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.
基金supported by grants from the National Major Project of the People's Republic of China (Nos. 2011ZX08002-004 and 2011ZX08010-004)the Wuhan Science and Technology Project (No. 201260523185)the International Scientific and Technological Cooperation Project of Ministry of Science and Technology of China (No. 2009DFB20290)
文摘Summary： Previous studies have shown that STAT3 plays a vital role in the genesis and progression of cancer. In this study, we investigated the relationship between the JAK2/STAT3 signalling pathway and germacrone-induced apoptosis in HepG2 cells. HepG2 cells were incubated with germacrone for 24 h, the protein expression of p-STAT3, STAT3, p-JAK2 and JAK2 was detected by Westem Blotting, and RT-PCR was used to determine the expression of STAT3, p53, Bcl-2 and Bax at transcriptional levels. Besides that, HepG2 cells were pre-treated with AG490 or IL-6 for 2 h, and then incubated with ger- macrone for 24 h. The expression ofp-JAK2, JAK2, p-STAT3, STAT3, p53, Bax and Bcl-2 was detected by Western blotting. The activity of HepG2 cells was tested by MTT assay. The apoptosis of HepG2 cells and levels of reactive oxygen species （ROS） were flow cytometrically measured. The results showed that germacrone exposure decreased p-STAT3 and p-JAK2 and regulated expression of p53 and Bcl-2 family members at the same time. Moreover, IL-6 enhanced the activation of the JAK2/STAT3 signalling pathway and therefore attenuated the germacrone-induced apoptosis. Suppression of JAK2/STAT3 signalling pathway by AG490, an inhibitor of JAK2, resulted in apoptosis and an increase in ROS in response to germacrone exposure. We therefore conclude that germacrone induces apoptosis through the JAK2/STAT3 signalling pathway.
基金supported by the Major State Basic Research Development Program (No.2008CB418102)the Environmental Monitoring Research Foundation of Jiangsu Province (No.0710)+1 种基金the Innovation Foundation for Youth Scholars of the State Key Laboratory of Pollution Control and Resource Reuse (No.PCRREF07002)the Specialized Research Fund for the Doctoral Program of Higher Education, SRFDP (No.200802841030)
文摘Water pollution of the Yangtze River in China became one of challenges that the government is facing today. Increasing numbers of petrochemical plants were built along the river in past decades, and numbers of organic chemicals were discharged into the river. Our goal was to establish in vitro system on rat sertoli cells, spermatogenic cells and leydig cells to investigate the reproductive toxicity potential induced by organic extracts from petrochemical effluents. Our results showed that the organic extract depressed the viability （p 〈 0.01） and destroyed the plasma membrane integrity of sertoli cells and spermatogenic cells to a certain degree. Accordingly, proportion of early apoptotic sertoli cells and late apoptotic spermatogenic cells increased significantly. Although significant morphological changes were not detected for leydig cells, the extract was observed to inhibit their testosterone production （p 〈 0.01）. Sertoli cells and sperrnatogenic cells appeared to be more sensitive and maybe the main targets of the key toxins. These results suggested that the in vitro system on rat testicular cells may be useful to predicate reproductive toxicity potential of organic extracts from petrochemical effluents. More attention should be paid to the petrochemical effluents, because long-term accumulation of these organic compounds in organisms may cause spermatogenesis malfunction and testosterone reduction.
基金supported by the grants from the Ministry of Science and Technology of China(Nos.2011CB965001 and 2011CB710905)the Knowledge Innovation Program of the Chinese Academy of Sciences(Nos.KSCX2-YW-R-232, KJCX2-YW-L08 and KYQY-QN-015)
文摘Mesenchymal stem cells （MSCs） show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities. MSCs are generally cultured on two-dimensional （2D） substrate in vitro. There are indications that they may simultaneously lose their sternness and multipotentiality as the result of prolonged 2D culture. In this study, we used three-dimensional （3D） collagen scaffolds as rat MSCs cartier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs. The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation. More importantly, compared to MSCs under 2D culture, 3D MSCs significantly maintained higher expression levels of stemness genes （Oct4, Sox2, Rex-1 and Nanog）, yielded high frequencies of colony-forming units-fibroblastic （CFU-F） and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction. Thus, 3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.
基金supported by grants from the Natural Science Foundation of Shandong Province of China (No. Y2007C102)the Medical Science and Technology Development Foundation of Shandong Province of China (No. 2007H2071)
文摘The expression and implication of vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （MMP-9） in residual hepatic tumor cells after lipiodol embolization were investi- gated. Two weeks after transplantation of VX2 tumor cells into the livers of rabbits, a xenograft model of the human hepatic neoplasm was successfully established. Forty rabbits were randomly divided into control group （n=20） and lipiodol group （n=20）. For the control group, 1 mL normal saline was injected through the gastroduodenal artery, whereas 0.3 mL/kg lipiodol was applied for the lipiodol group. One week after embolization, the expression level of VEGF in the plasma was measured by using en- zyme-linked immunosorbent assay （ELISA）. A three-step immunohistochemieal technique （ABC） was employed to detect the protein levels of VEGF and MMP-9 and the quantitative PCR for their mRNA levels was performed in the residual tumor cells. The VEGF in the plasma was significantly higher in the lipiodol group （1.42~0.29 ng/mL） than in the control group （1.12~0.21 ng/mL） （P〈0.01）. Moreover, the positive rate of VEGF protein in the residual tumor cells was significantly higher in the lipiodol group （62.13%~7.69%） than in the control group （53.16%~9.17%） （P〈0.05）. Similarly, the MMP-9 ex- pression in the residual ~mor cells was higher in the lipiodol group. The mRNA levels of VEGF （2.9313~2.4231） and MMP-9 （3.5721~1.6107） in the lipiodol group were significantly higher than those in the control group （1.5728~0.9453 and 1.7573~1.0641, respectively, P〈0.05）. Therefore, it was rea- sonable to speculate that the increased expression of VEGF and MMP-9 in residual hepatic tumor cells and tumor angiogenesis post-embolization would be responsible for the increased metastatic potentiality and invasiveness of these cells.
文摘Circulating tumor cells （CTCs） arise from primary or secondary tumors and enter the bloodstream by active or passive intravasation. Given the low number of CTCs, enrichment is necessary for detection. Filtration methods are based on selection of CTCs by size using a filter with 6.5 to 8 pm pores. After coloration, collected CTCs are evaluated according to morphological criteria. Immunophenotyping and fluorescence in situ hybridization techniques may be used. Selected CTCs can also be cultivated in vitro to provide more material. Analysis of genomic mutations is difficult because it requires adapted techniques due to limited DNA materials. Filtration-selected CTCs have shown prognostic value in many studies but multicentric validating trials are mandatory to strengthen this assessment. Other clinical applications are promising such as follow-up, therapy response prediction and diagnosis. Microfluidic emerging systems could optimize filtration-selected CTCs by increasing selection accuracy.
基金supported by the National Basic Research Program of China(Program 973)(Nos.2009CB941002 and 2011CB944202)the Distinguished Young Scholar Foundation of Heilongjiang Province(No.JC200905)
文摘Embryonic germ （EG） cells are cultured pluripotent stem cells derived from the primordial germ cells （PGCs） that migrate from the dorsal mesentery of the hindgut to the developing genital ridge. In this study, the morphology of the porcine genital ridge was assessed in embryos harvested on days 22-30 of pregnancy. PGCs from embryos at these stages were cultured to obtain porcine EG cell lines, and EG-like cells were derived from PGCs from embryos harvested on days 24-28 of pregnancy. The EG-like cells expressed Oct4, Sox2, Nanog, SSEA-3, SSEA-4 and alkaline phosphatase （AP）. These cells were able to form embryoid bodies （EBs） in suspension culture and differentiate into cells representative of the three germ layers as verified by a-fetoprotein （AFP）, s-smooth muscle actin （^-SMA）, and Nestin expression. Spontaneous differentiation from the porcine EG-like cells of delayed passage in vitro showed that they could differentiate into epithelial-like cells, mesenchymal-like cells and neuron-like cells. In vitro directed differentiation generated osteocytes, adipocytes and a variety of neural lineage cells, as demonstrated by alizarin red staining, oil red O staining, and immunoftuorescence for neuronal class III [3-tubulin （Tuj 1）, glial fibrillary protein （GFAP） and galactosylceramidase （GALC）, respectively. These results indicate that porcine EG-like cells have the potential for multi-lineage differentiation and are useful for basic porcine stem cell research.
基金the Guangdong Province Health Department (No. B30202)Guangzhou City Science and Technology Administration (No. 2006Z1-E0021)+1 种基金partly supported by the Guangdong Province Science and Technology Administration (No. 2007A032100003)Guangdong Provincial Medical Research Fund (No. A2008287)
文摘Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.
基金supported by the Fundamental Research Funds of the Central Universities(No.21613316)
文摘Estrogen-related receptor alpha （ERRα） plays an important role in the development of hor- monezdependent cancers, but its roles in lung cancer remain elusive. The present study was aimed to investigate the effects of ERRα on the proliferation and metastasis of lung cancer A549 cells. The mRNA and protein levels of ERRor were detected in lung cancer A549 and MCF-7 cells and bronchial epithelial BEAS-2B cells by qRT-PCR and Western blotting, respectively. ERRor plasmid transfection and XCT-790 （an inverse agonist of ERRc0 were used to up-regulate or down-regulate ERRα expression in A549 cells, respectively. The viability of A549 cells was measured by cell counting kit-8 （CCK-8） and the motility of A549 cells by wound healing assay and Transwell migration/invasion assay. The epithelial markers E-cadherin （E-Cad） and zona occludin-1 （ZO-1）, the mesenchymal markers fi- bronectin （FN） and vimentin （Vim） and the transcription factors （Snail, Zebl Twist and Slug） were fur- ther detected at mRNA and protein levels by qRT-PCR and Western blotting, respectively. The results showed that ERRor promoted the growth of lung cancer A549 cells in vitro. XCT-790 significantly in- hibited the migration and invasion of A549 cells. Over-expression of ERRα promoted the epithe- lial-to-mesenchymal transition （EMT） of A549 ceils, down-regulated the epithelial makers E-Cad and ZO-1, and up-regulated the mesenchymal makers FN and Vim. Silencing of Slug, but not other tran- scription factors, significantly abolished the ERRs-induced EMT of A549 cells. It was suggested that ERRor promoted the migration and invasion of A549 cells by inducing EMT, and Slug was involved in the process. Targeting ERRor might be an efficient approach for lung cancer treatment.
基金supported by the Postdoctoral Science Foundation of China, No. 20090461435
文摘Cultured Schwann cells were treated with 5.6 mM and 50 mM glucose alternating every 8 hours to simulate intermittent high glucose. The present study analyzed the neuroprotective effects of 1, 10 and 100 μM ginsenoside Rbl on oxidative damage and apoptosis in Schwann cells induced by intermittent high glucose. Flow cytometry demonstrated that ginsenoside Rbl reduced intermittent high glucose-mediated reactive oxygen species production. Enzyme linked immunosorbent assay showed that 8-hydroxy-2-deoxy guanosine levels in Schwann cells decreased following ginsenoside Rbl treatment. Quantitative real-time reverse transcription-PCR and western blot assay results revealed that ginsenoside Rbl inhibited intermittent high glucose-upregulated Bax expression, but antagonized intermittent high glucose-downregulated Bcl-2 expression in Schwann cells. These effects were most pronounced with 100 μM ginsenoside Rbl. These results indicate that ginsenoside Rbl inhibits intermittent high glucose-induced oxidative stress and apoptosis in Schwann cells.
