AIM: Ursolic acid (UA) and oleanolic acid (OA) are triperpene acids having a similar chemical structure and are distributed wildly in plants all over the world. In recent years, it was found that they had marked anti-...AIM: Ursolic acid (UA) and oleanolic acid (OA) are triperpene acids having a similar chemical structure and are distributed wildly in plants all over the world. In recent years, it was found that they had marked anti-tumor effects. There is little literature currently available regarding their effects on colon carcinoma cells. The present study was designed to investigate their inhibitory effects on human colon carcinoma cell line HCT15. METHODS: HCT15 cells were cultured with different drugs. The treated cells were stained with hematoxylin-eosin and their morphologic changes observed under a light microscope. The cytotoxicity of these drugs was evaluated by tetrazolium dye assay. Cell cycle analysis was performed by flow cytometry (FCM). Data were expressed as means +/-SEM and Analysis of variance and Student' t-test for individual comparisons. RESULTS: Twenty-four to 72 h after UA or OA 60 micromol/L treatment, the numbers of dead cells and cell fragments were increased and most cells were dead at the 72nd hour. The cytotoxicity of UA was stronger than that of OA. Seventy-eight hours after 30 micromol/L of UA or OA treatment, a number of cells were degenerated, but cell fragments were rarely seen. The IC(50) values for UA and OA were 30 and 60 micromol/L, respectively. Proliferation assay showed that proliferation of UA and OA-treated cells was slightly increased at 24h and significantly decreased at 48 h and 60 h, whereas untreated control cells maintained an exponential growth curve. Cell cycle analysis by FCM showed HCT15 cells treated with UA 30 and OA 60 for 36 h and 72 h gradually accumulated in G(0)/G(1) phase (both drugs P【0.05 for 72 h), with a concomitant decrease of cell populations in S phase (both drugs P【0.01 for 72 h) and no detectable apoptotic fraction. CONCLUSION: UA and OA have significant anti-tumor activity. The effect of UA is stronger than that of OA. The possible mechanism of action is that both drugs have an inhibitory effect on tumor cell proliferation through cell-cycle arrest.展开更多
Based on hourly precipitation data in eastern China in the warm season during 1961-2000,spatial distributions of frequency for 20 mm h 1 and 50 mm h 1 precipitation were analyzed,and the criteria of short-duration rai...Based on hourly precipitation data in eastern China in the warm season during 1961-2000,spatial distributions of frequency for 20 mm h 1 and 50 mm h 1 precipitation were analyzed,and the criteria of short-duration rainfall events and severe rainfall events are discussed.Furthermore,the percentile method was used to define local hourly extreme precipitation;based on this,diurnal variations and trends in extreme precipitation were further studied.The results of this study show that,over Yunnan,South China,North China,and Northeast China,the most frequent extreme precipitation events occur most frequently in late afternoon and/or early evening.In the Guizhou Plateau and the Sichuan Basin,the maximum frequency of extreme precipitation events occurs in the late night and/or early morning.And in the western Sichuan Plateau,the maximum frequency occurs in the middle of the night.The frequency of extreme precipitation （based on hourly rainfall measurements） has increased in most parts of eastern China,especially in Northeast China and the middle and lower reaches of the Yangtze River,but precipitation has decreased significantly in North China in the past 50 years.In addition,stations in the Guizhou Plateau and the middle and lower reaches of the Yangtze River exhibit significant increasing trends in hourly precipitation extremes during the nighttime more than during the daytime.展开更多
Recent advances in studies on the interaction between the East Asian monsoon and the ENSO cycle are reviewed in this paper. Through the recent studies, not only have the responding features and processes of the East A...Recent advances in studies on the interaction between the East Asian monsoon and the ENSO cycle are reviewed in this paper. Through the recent studies, not only have the responding features and processes of the East Asian winter and summer monsoon circulation anomalies and summer rainfall anomalies in East Asia to the ENSO cycle during its different stages been understood further, but also have the thermal and dynamic effects of the tropical \vestern Pacific on the ENSO cycle been deeply analyzed from the observational facts and dynamic theories. The results of observational and theoretical studies showed that the dynamical effect of the atmospheric circulation and zonai wind anomalies in the lower troposphere over the tropical western Pacific on the ENSO cycle may be through the excitation of the equatorial oceanic Kelvin wave and Rossby waves in the equatorial Pacific. These studies demonstrated further that the ENSO cycle originates from the tropical western Pacific. Moreover, these recent studies also showed that the atmospheric circulation and zonai wind anomalies over the tropical western Pacific not only result from the air-sea interaction over the tropical western Pacific, but are also greatly influenced by the East Asian winter and summer monsoons. Additionally, the scientific problems in the interaction between the Asian monsoon and the ENSO cycle which should be studied further in the near future are also pointed out in this paper.展开更多
The projected changes in carbon exchange between China terrestrial ecosystem and the atmosphere and vegetation and soil carbon storage during the 21st century were investigated using an atmos-phere-vegetation interact...The projected changes in carbon exchange between China terrestrial ecosystem and the atmosphere and vegetation and soil carbon storage during the 21st century were investigated using an atmos-phere-vegetation interaction model (AVIM2). The results show that in the coming 100 a, for SRES B2 scenario and constant atmospheric CO2 concentration, the net primary productivity (NPP) of terrestrial ecosystem in China will be decreased slowly, and vegetation and soil carbon storage as well as net ecosystem productivity (NEP) will also be decreased. The carbon sink for China terrestrial ecosystem in the beginning of the 20th century will become totally a carbon source by the year of 2020, while for B2 scenario and changing atmospheric CO2 concentration, NPP for China will increase continuously from 2.94 GtC·a?1 by the end of the 20th century to 3.99 GtC·a?1 by the end of the 21st century, and vegetation and soil carbon storage will increase to 110.3 GtC. NEP in China will keep rising during the first and middle periods of the 21st century, and reach the peak around 2050s, then will decrease gradually and approach to zero by the end of the 21st century.展开更多
Hepatocellular carcinoma(HCC)is an aggressive human cancer with increasing incidence worldwide.Multiple efforts have been made to explore pharmaceutical therapies to treat HCC,such as targeted tyrosine kinase inhibito...Hepatocellular carcinoma(HCC)is an aggressive human cancer with increasing incidence worldwide.Multiple efforts have been made to explore pharmaceutical therapies to treat HCC,such as targeted tyrosine kinase inhibitors,immune based therapies and combination of chemotherapy.However,limitations exist in current strategies including chemoresistance for instance.Tumor initiation and progression is driven by reprogramming of metabolism,in particular during HCC development.Recently,metabolic associated fatty liver disease(MAFLD),a reappraisal of new nomenclature for nonalcoholic fatty liver disease(NAFLD),indicates growing appreciation of metabolism in the pathogenesis of liver disease,including HCC,thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment.In this review,we introduce directions by highlighting the metabolic targets in glucose,fatty acid,amino acid and glutamine metabolism,which are suitable for HCC pharmaceutical intervention.We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment.Furthermore,opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.展开更多
The aim of this study is to assess the effects of DNA methylation and historic acetylation, alone or in combination, on the expression of several tumor-associated genes and cell cycle progression in two established hu...The aim of this study is to assess the effects of DNA methylation and historic acetylation, alone or in combination, on the expression of several tumor-associated genes and cell cycle progression in two established human colon cancer cell lines: Colo-320 and SW1116. Treatments with 5-aza-2'-deoxycytidine (5-aza-dC) and trichostatin A, alone or in combination, were applied respectively. The methylation status of the CDKN2A promoter was determined by methyla-tion-specific PCR, and the acetylated status of the histones associated with the p21WAF1 and CDKN2A genes was examined by chromatin immunoprecipitation. The expression of the CDKN2A, p21WAF1, p53, p73, APC, c-myc, c-Ki-ras and survivin genes was detected by real-time RT-PCR and RT-PCR. The cell cycle profile was established by flow cytometry. We found that along with the demethylation of the CDKN2A gene promoter in both cell lines induced by 5-aza-dC alone or in combination with TSA, the expression of both CDKN2A and APC genes increased. The treatment of TSA or sodium butyrate up-regulated the transcription of p21WAF1 significantly by inducing the acetylation of histones H4 and H3, but failed to alter the acetylation level of CDKN2A-associated histones. No changes in transcription of p53, p73, c-myc, c-Ki-ras and survivin genes were observed. In addition, TSA or sodium butyrate was shown to arrest cells at the G1 phase. However, 5-aza-dC was not able to affect the cell cycle progression. In conclusion, regulation by epigenetic modification of the transcription of tumor-associated genes and the cell cycle progression in both human colon cancer cell lines Colo-320 and SW1116 is gene-specific.展开更多
Vascular smooth muscle cell （VSMC） differentiation and proliferation are two important physiological proc- esses during vascular development. The phenotypic alteration from differentiated to proliferative VSMC contr...Vascular smooth muscle cell （VSMC） differentiation and proliferation are two important physiological proc- esses during vascular development. The phenotypic alteration from differentiated to proliferative VSMC contrib- utes to the development of several major cardiovascular diseases including atherosclerosis, hypertension, resteno- sis after angioplasty or bypass, diabetic vascular complications, and transplantation arteriopathy. Since the VSMC phenotype in these pathological conditions resembles that of developing VSMC during embryonic development, understanding of the molecular mechanisms that control VSMC differentiation will provide fundamental insights into the pathological processes of these cardiovascular diseases. Although VSMC differentiation is usually ac- companied by an irreversible cell cycle exit, VSMC proliferation and differentiation occur concurrently during embryonic development. The molecular mechanisms simultaneously regulating these two processes, however, remain largely unknown. Our recent study demonstrates that cell division cycle 7, a key regulator of cell cycle, promotes both VSMC differentiation and proliferation through different mechanisms during the initial phase of VSMC differentiation. Conversely, Kriappel-like factor 4 appears to be a repressor for both VSMC differentia- tion and proliferation. This review attempts to highlight the novel role of cell division cycle 7 in TGF-β-induced VSMC differentiation and proliferation. The role of K141ppel-like factor 4 in suppressing these two processes will also be discussed.展开更多
Guliya ice core records, high lake-level records in the Qinghai-Xizang Plateau and at its north side as well as vegetation succession records indicated that during the period of 30-40 kaBP, namely the later age of the...Guliya ice core records, high lake-level records in the Qinghai-Xizang Plateau and at its north side as well as vegetation succession records indicated that during the period of 30-40 kaBP, namely the later age of the megainterstadial of last glacial period, or the marine oxygen isotope stage 3, the climate of the Qinghai-Xizang Plateau was exceptionally warm and humid, the temperature was 2-4℃ higher than today and the precipitation was 40% to over 100%展开更多
Enhancing the terrestrial ecosystem carbon sink(referred to as terrestrial C sink) is an important way to slow down the continuous increase in atmospheric carbon dioxide(CO_(2)) concentration and to achieve carbon neu...Enhancing the terrestrial ecosystem carbon sink(referred to as terrestrial C sink) is an important way to slow down the continuous increase in atmospheric carbon dioxide(CO_(2)) concentration and to achieve carbon neutrality target.To better understand the characteristics of terrestrial C sinks and their contribution to carbon neutrality,this review summarizes major progress in terrestrial C budget researches during the past decades,clarifies spatial patterns and drivers of terrestrial C sources and sinks in China and around the world,and examines the role of terrestrial C sinks in achieving carbon neutrality target.According to recent studies,the global terrestrial C sink has been increasing from a source of (-0.2±0.9) Pg C yr^(-1)(1 Pg=1015g)in the 1960s to a sink of (1.9±1.1) Pg C yr^(-1) in the 2010s.By synthesizing the published data,we estimate terrestrial C sink of 0.20–0.25 Pg C yr^(-1) in China during the past decades,and predict it to be 0.15–0.52 Pg C yr^(-1) by 2060.The terrestrial C sinks are mainly located in the mid-and high latitudes of the Northern Hemisphere,while tropical regions act as a weak C sink or source.The C balance differs much among ecosystem types:forest is the major C sink;shrubland,wetland and farmland soil act as C sinks;and whether the grassland functions as C sink or source remains unclear.Desert might be a C sink,but the magnitude and the associated mechanisms are still controversial.Elevated atmospheric CO_(2) concentration,nitrogen deposition,climate change,and land cover change are the main drivers of terrestrial C sinks,while other factors such as fires and aerosols would also affect ecosystem C balance.The driving factors of terrestrial C sink differ among regions.Elevated CO_(2) concentration and climate change are major drivers of the C sinks in North America and Europe,while afforestation and ecological restoration are additionally important forcing factors of terrestrial C sinks in China.For future studies,we recommend the necessity for intensive and long-term ec展开更多
文摘AIM: Ursolic acid (UA) and oleanolic acid (OA) are triperpene acids having a similar chemical structure and are distributed wildly in plants all over the world. In recent years, it was found that they had marked anti-tumor effects. There is little literature currently available regarding their effects on colon carcinoma cells. The present study was designed to investigate their inhibitory effects on human colon carcinoma cell line HCT15. METHODS: HCT15 cells were cultured with different drugs. The treated cells were stained with hematoxylin-eosin and their morphologic changes observed under a light microscope. The cytotoxicity of these drugs was evaluated by tetrazolium dye assay. Cell cycle analysis was performed by flow cytometry (FCM). Data were expressed as means +/-SEM and Analysis of variance and Student' t-test for individual comparisons. RESULTS: Twenty-four to 72 h after UA or OA 60 micromol/L treatment, the numbers of dead cells and cell fragments were increased and most cells were dead at the 72nd hour. The cytotoxicity of UA was stronger than that of OA. Seventy-eight hours after 30 micromol/L of UA or OA treatment, a number of cells were degenerated, but cell fragments were rarely seen. The IC(50) values for UA and OA were 30 and 60 micromol/L, respectively. Proliferation assay showed that proliferation of UA and OA-treated cells was slightly increased at 24h and significantly decreased at 48 h and 60 h, whereas untreated control cells maintained an exponential growth curve. Cell cycle analysis by FCM showed HCT15 cells treated with UA 30 and OA 60 for 36 h and 72 h gradually accumulated in G(0)/G(1) phase (both drugs P【0.05 for 72 h), with a concomitant decrease of cell populations in S phase (both drugs P【0.01 for 72 h) and no detectable apoptotic fraction. CONCLUSION: UA and OA have significant anti-tumor activity. The effect of UA is stronger than that of OA. The possible mechanism of action is that both drugs have an inhibitory effect on tumor cell proliferation through cell-cycle arrest.
基金supported by the R & D Special Fund for Public Welfare Industry (meteorology)(GYHY201106018)National Key Program for Developing Basic Sciences (Grant No. 2006CB400503)
文摘Based on hourly precipitation data in eastern China in the warm season during 1961-2000,spatial distributions of frequency for 20 mm h 1 and 50 mm h 1 precipitation were analyzed,and the criteria of short-duration rainfall events and severe rainfall events are discussed.Furthermore,the percentile method was used to define local hourly extreme precipitation;based on this,diurnal variations and trends in extreme precipitation were further studied.The results of this study show that,over Yunnan,South China,North China,and Northeast China,the most frequent extreme precipitation events occur most frequently in late afternoon and/or early evening.In the Guizhou Plateau and the Sichuan Basin,the maximum frequency of extreme precipitation events occurs in the late night and/or early morning.And in the western Sichuan Plateau,the maximum frequency occurs in the middle of the night.The frequency of extreme precipitation （based on hourly rainfall measurements） has increased in most parts of eastern China,especially in Northeast China and the middle and lower reaches of the Yangtze River,but precipitation has decreased significantly in North China in the past 50 years.In addition,stations in the Guizhou Plateau and the middle and lower reaches of the Yangtze River exhibit significant increasing trends in hourly precipitation extremes during the nighttime more than during the daytime.
基金supported by the National Natural Science Foundation of China(Grant No.40231005)the Progran for Knowledge Innovation Project of Chinese Academy of Sciences(No.KZCX3-SW-218,KZCX2-SW-210).
文摘Recent advances in studies on the interaction between the East Asian monsoon and the ENSO cycle are reviewed in this paper. Through the recent studies, not only have the responding features and processes of the East Asian winter and summer monsoon circulation anomalies and summer rainfall anomalies in East Asia to the ENSO cycle during its different stages been understood further, but also have the thermal and dynamic effects of the tropical \vestern Pacific on the ENSO cycle been deeply analyzed from the observational facts and dynamic theories. The results of observational and theoretical studies showed that the dynamical effect of the atmospheric circulation and zonai wind anomalies in the lower troposphere over the tropical western Pacific on the ENSO cycle may be through the excitation of the equatorial oceanic Kelvin wave and Rossby waves in the equatorial Pacific. These studies demonstrated further that the ENSO cycle originates from the tropical western Pacific. Moreover, these recent studies also showed that the atmospheric circulation and zonai wind anomalies over the tropical western Pacific not only result from the air-sea interaction over the tropical western Pacific, but are also greatly influenced by the East Asian winter and summer monsoons. Additionally, the scientific problems in the interaction between the Asian monsoon and the ENSO cycle which should be studied further in the near future are also pointed out in this paper.
基金the Basic Research Program of China (Grant No. 2002CB412500)the National Natural Science Foundation of China (Grant No. 30590384)
文摘The projected changes in carbon exchange between China terrestrial ecosystem and the atmosphere and vegetation and soil carbon storage during the 21st century were investigated using an atmos-phere-vegetation interaction model (AVIM2). The results show that in the coming 100 a, for SRES B2 scenario and constant atmospheric CO2 concentration, the net primary productivity (NPP) of terrestrial ecosystem in China will be decreased slowly, and vegetation and soil carbon storage as well as net ecosystem productivity (NEP) will also be decreased. The carbon sink for China terrestrial ecosystem in the beginning of the 20th century will become totally a carbon source by the year of 2020, while for B2 scenario and changing atmospheric CO2 concentration, NPP for China will increase continuously from 2.94 GtC·a?1 by the end of the 20th century to 3.99 GtC·a?1 by the end of the 21st century, and vegetation and soil carbon storage will increase to 110.3 GtC. NEP in China will keep rising during the first and middle periods of the 21st century, and reach the peak around 2050s, then will decrease gradually and approach to zero by the end of the 21st century.
基金supported by the National Natural Science Foundation of China(No.82070883)Scientific Research Foundation for high-level faculty,China Pharmaceutical University(Nanjing,China)。
文摘Hepatocellular carcinoma(HCC)is an aggressive human cancer with increasing incidence worldwide.Multiple efforts have been made to explore pharmaceutical therapies to treat HCC,such as targeted tyrosine kinase inhibitors,immune based therapies and combination of chemotherapy.However,limitations exist in current strategies including chemoresistance for instance.Tumor initiation and progression is driven by reprogramming of metabolism,in particular during HCC development.Recently,metabolic associated fatty liver disease(MAFLD),a reappraisal of new nomenclature for nonalcoholic fatty liver disease(NAFLD),indicates growing appreciation of metabolism in the pathogenesis of liver disease,including HCC,thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment.In this review,we introduce directions by highlighting the metabolic targets in glucose,fatty acid,amino acid and glutamine metabolism,which are suitable for HCC pharmaceutical intervention.We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment.Furthermore,opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.
基金This work was supported in part by National Natural Science Foundation of China(No.30170413)the Foundation for Jing Yuan FANG of National Excellent Doctoral Dissertation of China(No.199946)the Foundation of Shanghai Education Committee(Shuguang Plan,No.02SG45).
文摘The aim of this study is to assess the effects of DNA methylation and historic acetylation, alone or in combination, on the expression of several tumor-associated genes and cell cycle progression in two established human colon cancer cell lines: Colo-320 and SW1116. Treatments with 5-aza-2'-deoxycytidine (5-aza-dC) and trichostatin A, alone or in combination, were applied respectively. The methylation status of the CDKN2A promoter was determined by methyla-tion-specific PCR, and the acetylated status of the histones associated with the p21WAF1 and CDKN2A genes was examined by chromatin immunoprecipitation. The expression of the CDKN2A, p21WAF1, p53, p73, APC, c-myc, c-Ki-ras and survivin genes was detected by real-time RT-PCR and RT-PCR. The cell cycle profile was established by flow cytometry. We found that along with the demethylation of the CDKN2A gene promoter in both cell lines induced by 5-aza-dC alone or in combination with TSA, the expression of both CDKN2A and APC genes increased. The treatment of TSA or sodium butyrate up-regulated the transcription of p21WAF1 significantly by inducing the acetylation of histones H4 and H3, but failed to alter the acetylation level of CDKN2A-associated histones. No changes in transcription of p53, p73, c-myc, c-Ki-ras and survivin genes were observed. In addition, TSA or sodium butyrate was shown to arrest cells at the G1 phase. However, 5-aza-dC was not able to affect the cell cycle progression. In conclusion, regulation by epigenetic modification of the transcription of tumor-associated genes and the cell cycle progression in both human colon cancer cell lines Colo-320 and SW1116 is gene-specific.
基金supported by grants from National Institutes of Health (HL093429 and HL107526 to S.-Y.C.)
文摘Vascular smooth muscle cell （VSMC） differentiation and proliferation are two important physiological proc- esses during vascular development. The phenotypic alteration from differentiated to proliferative VSMC contrib- utes to the development of several major cardiovascular diseases including atherosclerosis, hypertension, resteno- sis after angioplasty or bypass, diabetic vascular complications, and transplantation arteriopathy. Since the VSMC phenotype in these pathological conditions resembles that of developing VSMC during embryonic development, understanding of the molecular mechanisms that control VSMC differentiation will provide fundamental insights into the pathological processes of these cardiovascular diseases. Although VSMC differentiation is usually ac- companied by an irreversible cell cycle exit, VSMC proliferation and differentiation occur concurrently during embryonic development. The molecular mechanisms simultaneously regulating these two processes, however, remain largely unknown. Our recent study demonstrates that cell division cycle 7, a key regulator of cell cycle, promotes both VSMC differentiation and proliferation through different mechanisms during the initial phase of VSMC differentiation. Conversely, Kriappel-like factor 4 appears to be a repressor for both VSMC differentia- tion and proliferation. This review attempts to highlight the novel role of cell division cycle 7 in TGF-β-induced VSMC differentiation and proliferation. The role of K141ppel-like factor 4 in suppressing these two processes will also be discussed.
文摘Guliya ice core records, high lake-level records in the Qinghai-Xizang Plateau and at its north side as well as vegetation succession records indicated that during the period of 30-40 kaBP, namely the later age of the megainterstadial of last glacial period, or the marine oxygen isotope stage 3, the climate of the Qinghai-Xizang Plateau was exceptionally warm and humid, the temperature was 2-4℃ higher than today and the precipitation was 40% to over 100%
基金supported by the National Natural Science Foundation of China (31988102)。
文摘Enhancing the terrestrial ecosystem carbon sink(referred to as terrestrial C sink) is an important way to slow down the continuous increase in atmospheric carbon dioxide(CO_(2)) concentration and to achieve carbon neutrality target.To better understand the characteristics of terrestrial C sinks and their contribution to carbon neutrality,this review summarizes major progress in terrestrial C budget researches during the past decades,clarifies spatial patterns and drivers of terrestrial C sources and sinks in China and around the world,and examines the role of terrestrial C sinks in achieving carbon neutrality target.According to recent studies,the global terrestrial C sink has been increasing from a source of (-0.2±0.9) Pg C yr^(-1)(1 Pg=1015g)in the 1960s to a sink of (1.9±1.1) Pg C yr^(-1) in the 2010s.By synthesizing the published data,we estimate terrestrial C sink of 0.20–0.25 Pg C yr^(-1) in China during the past decades,and predict it to be 0.15–0.52 Pg C yr^(-1) by 2060.The terrestrial C sinks are mainly located in the mid-and high latitudes of the Northern Hemisphere,while tropical regions act as a weak C sink or source.The C balance differs much among ecosystem types:forest is the major C sink;shrubland,wetland and farmland soil act as C sinks;and whether the grassland functions as C sink or source remains unclear.Desert might be a C sink,but the magnitude and the associated mechanisms are still controversial.Elevated atmospheric CO_(2) concentration,nitrogen deposition,climate change,and land cover change are the main drivers of terrestrial C sinks,while other factors such as fires and aerosols would also affect ecosystem C balance.The driving factors of terrestrial C sink differ among regions.Elevated CO_(2) concentration and climate change are major drivers of the C sinks in North America and Europe,while afforestation and ecological restoration are additionally important forcing factors of terrestrial C sinks in China.For future studies,we recommend the necessity for intensive and long-term ec
