Myocardial ischemia is a serious threat to human health,and vascular dysfunction is its main cause.Buxu Tongyu(BXTY)Granule is an effective traditional Chinese medicine(TCM)for treating myocardial ischemia.However,the...Myocardial ischemia is a serious threat to human health,and vascular dysfunction is its main cause.Buxu Tongyu(BXTY)Granule is an effective traditional Chinese medicine(TCM)for treating myocardial ischemia.However,the underlying mechanism of BXTY is still unclear.In this study,we demonstrate that BXTY ameliorates myocardial ischemia by activating the soluble guanylate cyclase(sGC)-30,50-cyclic guanosine monophosphate(cGMP)-protein kinase G(PKG)signaling pathway in vascular smooth muscle cells(VSMCs)to dilate the arteries.BXTY was given by gavage for ten consecutive days before establishing an animal model of acute myocardial ischemia in mice via the intraperitoneal injection of pituitrin.The results showed that BXTY alleviated the symptoms of myocardial ischemia induced by pituitrin in mice,including electrocardiogram abnormalities and changes in plasma enzymes.In addition,BXTY dilated pre-constricted blood vessels and inhibited the vasoconstriction of the superior mesenteric artery in a dose-dependent but endothelial-independent manner.These effects were eliminated by preincubating vascular rings with the sGC inhibitors NS 2028 or ODQ,or with the PKG inhibitor KT 5823.Moreover,BXTY increased the protein expression of sGC-b1 and the intracellular second messenger cGMP level in mouse aortic vascular smooth muscle cells(MOVAs).NS 2028 or ODQ reversed these effects of BXTY.The expression level of the cGMP downstream effector protein PKG-1 increased after treating MOVAs with BXTY.NS 2028,ODQ,or KT 5823 also reversed this effect of BXTY.In conclusion,BXTY can improve the symptoms of acute myocardial ischemia in mice,and activating the sGC-cGMP-PKG pathway in VSMCs to induce vasodilation is its key pharmacodynamic mechanism.展开更多
The increasing incidence of cardiovascular disease(CVD)is a significant global health concern,affecting millions of individuals each year.Accurate diagnosis of acute CVD poses a formidable challenge,as misdiagnosis ca...The increasing incidence of cardiovascular disease(CVD)is a significant global health concern,affecting millions of individuals each year.Accurate diagnosis of acute CVD poses a formidable challenge,as misdiagnosis can significantly decrease patient survival rates.Traditional biomarkers have played a vital role in the diagnosis and prognosis of CVDs,but they can be influenced by various factors,such as age,sex,and renal function.Soluble ST2(sST2)is a novel biomarker that is closely associated with different CVDs.Its low reference change value makes it suitable for continuous measurement,unaffected by age,kidney function,and other confounding factors,facilitating risk stratification of CVDs.Furthermore,the combination of sST2 with other biomarkers can enhance diagnostic accuracy and prognostic value.This review aims to provide a comprehensive overview of sST2,focusing on its diagnostic and prognostic value as a myocardial marker for different types of CVDs and discussing the current limitations of sST2.展开更多
Solubility enhancement has been a priority to overcome poor solubility with optoelectronic molecules for solution-processable devices. This study aims to obtain experimental data on the effect of particle sizes on the...Solubility enhancement has been a priority to overcome poor solubility with optoelectronic molecules for solution-processable devices. This study aims to obtain experimental data on the effect of particle sizes on the solubility properties of several typical optoelectronic molecules in organic solvents, including the solubility results of 1,3-bis(9-carbazolyl)benzene(m CP), 1,3,5-tris(1-phenyl-1H-benzimidazol-2-yl)ben zene(TPBi) and 2-(4-tert-butylphenyl)-5-(4-biphenyl)-1,3,4-oxadiazole(PBD) in ethanol and acetonitrile,respectively. Nanoparticles of m CP, TPBi and PBD with sizes from dozens to several hundred nanometers were prepared by solvent antisolvent precipitation method and their solubility were determined by using isothermal saturation method. The saturation solubility of nanoparticles of three kinds of optoelectronic molecules exhibited increase of 12.9%-25.7% in comparison to the same raw materials in the form of microparticles. The experimental evidence indicates that nanonization technology is a feasible way to make optoelectronic molecules dissolve in liquids with enhanced solubility.展开更多
To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chromatography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and q...To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chromatography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.Attributes of nanobody based pharmaceutical assays are discussed.展开更多
Objective:To construct a secretory eukaryotic expression vector of DSG2 fused with the Fc region of the human IgG,to validate its expression in 293T cells,and to purify the secretory protein with biological activity.M...Objective:To construct a secretory eukaryotic expression vector of DSG2 fused with the Fc region of the human IgG,to validate its expression in 293T cells,and to purify the secretory protein with biological activity.Methods:The DSG2 extracellular domain fragment gene(DSG2ex),was amplified by PCR,and was inserted into the eukaryotic expression plasmid pCMV3-IgG1 to construct the recombinant eukaryotic expression plasmid-pCMV3-DSG2ex-IgG1.The successfully constructed eukaryotic expression plasmid was transfected into 293T cells to express and secrete DSG2 extracellular domain protein.The targeted protein was purified from the cell culture supernatant by Protein A affinity chromatography and confirmed by Western Blotting and ELISA.Results:The pCMV3-DSG2ex-IgG1 eukaryotic expression plasmid was successfully constructed.The highest protein expression level was obtained with 293T cells after 96 h of transfection.The relative molecular mass of the purified product was between 100 and 130 kDa was estimated by SDS-PAGE,which was consistent with the expectation.The yield of the purified protein reached 0.8 mg/ml with a purity over 90%.The purified DSG2 extracellular domain protein with IgG1 tag was recognized by IgG monoclonal antibodies by Western blotting.Moreover,the ELISA results showed that the prepared DSG2 extracellular domain protein had significant binding activity to human type 55 adenovirus Fiber Knob protein(HAdV-55).Conclusion:A simple and efficient method for eukaryotic expression and purification of human soluble DSG2 extracellular domain protein was successfully established,and biologically active DSG2 extracellular domain protein was purified,which laid the foundation for the later study of its protein function and anti-adenovirus drugs.展开更多
BACKGROUND Several studies of spontaneous intracerebral hemorrhage(SICH)patients have shown apoptotic changes in brain samples after hematoma evacuation.However,there have been no data on the association between blood...BACKGROUND Several studies of spontaneous intracerebral hemorrhage(SICH)patients have shown apoptotic changes in brain samples after hematoma evacuation.However,there have been no data on the association between blood concentrations of soluble fas(sFas)(the main surface death receptor of the extrinsic apoptosis pathway)and the prognosis of spontaneous intracranial hypotension(SIH)patients.AIM To determine whether there is an association between blood sFas concentrations and SICH patient mortality.METHODS We included patients with severe and supratentorial SIH.Severe was defined as having Glasgow Coma Scale<9.We determined serum sFas concentrations at the time of severe SICH diagnosis.RESULTS We found that non-surviving patients(n=36)compared to surviving patients(n=39)had higher ICH score(P=0.001),higher midline shift(P=0.004),higher serum sFas concentrations(P<0.001),and lower rate of early hematoma evacuation(P=0.04).Multiple logistic regression analysis showed an association between serum sFas concentrations and 30-d mortality(odds ratio=1.070;95%confidence interval=1.014-1.129;P=0.01)controlling for ICH score,midline shift,and early hematoma evacuation.CONCLUSION The association of blood sFas concentrations and SICH patient mortality is a novel finding in our study.展开更多
Bai-Hua-She-She-Cao Hedyotis diffusa Willd. (Ru-biaceae) is a medicinal herbwidely distributed in northeast Asian countries. In traditional Chinese medicine, it has the effectof 'clearing away heat and toxic mater...Bai-Hua-She-She-Cao Hedyotis diffusa Willd. (Ru-biaceae) is a medicinal herbwidely distributed in northeast Asian countries. In traditional Chinese medicine, it has the effectof 'clearing away heat and toxic material, promoting blood circulation and removing blood stasis'.It is a well known Chinese folk-medicine used for the treatment of appendicitis, sore throat, mumps,acne, sebo-rheic dermatitis and various kinds of tumors, such as tumors of digestive tract,carcinoma of liver. It was reported that the MeOH extract of H. diffusa demonstrated a significantantitumor activity and ursolic acid succeeded in being isolated from the MeOH extract as an activecomponent . Shan BN, et al suggested that the direct aqueous extract of H. diffusa hadimmuno-modulating activity and antitumor activity in vitro through stimulating the immune system tokill or engulf tumor cells. But regarding anti-tumor activity in vivo of water soluble extracts fromH. diffusa, no detail was reported. Therefore, we prepared water soluble extracts (H_1 and H_2)from H. diffusa and evaluated their anti-tumor property in vivo experiments as well as protectiveeffect on chemo-therapeutic damage.展开更多
Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ ...Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3,and 0 05?mmol·L -1 of CaM antagonist CPZ. The changes in the contents of total soluble protein and CaM, and freezing resistance in all pretreated seedlings in various periods ( viz: following cold acclimation, chilling stress and recovery) were investigated. Results showed that cold acclimation increased the contents of total soluble protein and CaM, and freezing resistance of seedlings, which could be strongly reduced by the pretreatments of EGTA CPZ and LaCl 3 Cold acclimation combined with CaCl 2 pretreatment enhanced the effect of cold acclimation on freezing resistance, and obviously increased the contents of total soluble protein and CaM, reduced the declining degree of the contents of total soluble protein and CaM caused by chilling stress as compared with cold acclimation, augmented the increase in the level of total soluble protein and CaM during the recovery periods. Further analysis found that an increase in total soluble protein content during cold acclimation with or without CaCl 2 pretreatment mainly resulted from the increase in content of heat stable protein in total soluble protein. It is suggested that Ca 2+ calmodulin may be involved in the synthesis of total soluble protein, and the induction of freezing resistance of seedlings.展开更多
The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress an...The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress and the recovery after chilling stress), and the survival rate and the freezing resistance of cuttings during cold acclimation at -3℃ were investigated. Results showed that cold acclimation not only increased the contents of total soluble protein and RNA, the survival rates and the freezing resistance of cuttings, decreased the activity of RNase, but also reduced the declining degree of total soluble protein and RNA contents, and the increasing level of RNase caused by chilling stress as compared with the controls. In addition, cold acclimation augmented the increase in the level of total soluble protein and RNA, and facilitated the decrease of RNase during the recovery periods. Further analysis found that the DNA content of all treatments kept relative stability at various periods. The changes in total soluble protein, RNA and RNase were closely related to the freezing resistance of cuttings. It appears that the increase of RNA content caused by cold acclimation induced decrease of RNase activity may be involved in the accumulation of total soluble protein and the induction of freezing resistance of cuttings.展开更多
Pervoskite type oxides LaCoO 3 was prepared by citrate method with the granula of 20 nm-30 nm. With a fluorescent Hg lamp or sunlight as irradiator, the degradation experiments of various water soluble dyes we...Pervoskite type oxides LaCoO 3 was prepared by citrate method with the granula of 20 nm-30 nm. With a fluorescent Hg lamp or sunlight as irradiator, the degradation experiments of various water soluble dyes were carried out in the suspension system of LaCoO 3 . The results show that the pervoskite type oxide LaCoO 3 has good photocatalytic activity.Studied by X ray photoelectron spectroscopy and photoacoustic spectra, its photocatalytic activity is found to be related with factors such as the d electron structure of ion Co 3+ ,Co—O binding energy and adsorbed oxygen on the surface etc.展开更多
[Objective] The extraction efficiency of soluble nitrogen in common moun- tain red soil under different conditions was analyzed to provide experimental refer- ences for the extraction of soil soluble nitrogen. [Method...[Objective] The extraction efficiency of soluble nitrogen in common moun- tain red soil under different conditions was analyzed to provide experimental refer- ences for the extraction of soil soluble nitrogen. [Method] The fresh soil and air-dried soil, which were the common mountain red soil under evergreen broad-leaved forest vegetation, were used as the experimental materials. The 2 mol/L KCI and 0.5 mol/L K2SO4 were used as the extractants to extract the soil soluble nitrogen with the extractant-to-soil volume ratio at 4：1, 6：1 and 10：1, respectively, to examine the extrac- tion efficiency of soluble nitrogen in the experimental soil with different extraction conditions. [Result] Compared with K2SO4, the NH4＋-N extracted by KCI was better in quantity and stability; while only at high extractant-to-soil volume ratio could K2SO4 fully extract the soil NH4＋-N. The 2 extractants showed small difference in extracting NO3--N, but at low extractant-to-soil volume ratio, the stability of NO3--N extracted by KCI was higher than that extracted by K2SO4. The properties of NO2- -N extracted by the 2 extractants were very similar. As for soil SON extraction, there was small dif- ference between the 2 extractants. However, the SON extracted by KCI was fewer in quantity than that by K2SO4, but had higher stability, and the extracted SON amount of the 2 extractants showed significant correlation. The extractant-to-soil ratio had little effect on MBN extraction using K2SO4, but the effect was significant when using KCI to extract MBN. The difference between the extraction efficiency of KCI and K^SO4 in extracting fresh soils was greater than that in extracting air-dried soil; therefore, the extractants should be carefully selected in soluble nitrogen extraction from fresh soil. [Conclusion] The results lay the experimental foundation for improving the extraction efficiency of soil soluble nitrogen and increasing the comparability of measurement results.展开更多
Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of ex...Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of expressed proteins also varied amongst the samples. The fewest soluble proteins were present in filamentous sporophytes. There were more types and amounts of soluble protein in conchospores than in filamentous sporophytes, but fewer than in bulgy sporophytes. More types of protein were detected in filamentous sporophytes cultivated in high temperatures than in those growing in normal situations. The most types and amounts of protein were found in blade gametophytes in all samples. Blade gametophytes harvested last year and stored at -20 ℃ showed only minor differences in expression of proteins when compared with those harvested in different seasons.展开更多
[Objective] The research aimed to provide theoretical basis for the cultivation and production of peach.[Method] The three-year-old seedling peach tree was used as the materials,and NH4NO3 was used as the experimental...[Objective] The research aimed to provide theoretical basis for the cultivation and production of peach.[Method] The three-year-old seedling peach tree was used as the materials,and NH4NO3 was used as the experimental nitrogen fertilizer.Three nitrogen levels,0,3,6 g per pot respectively were set to study the effects of different nitrogen fertilizer on the contents of soluble sugar,starch and internal structure of the root of the peaches.[Result] The contents of soluble sugar of roots and leaves increased with the increase of nitrogen fertilizer level,and the level of 6 g per pot was the highest,which showed extreme differences between the level of 3 g per pot and the control.However,starch contents showed opposite results,which decreased as the level of nitrogen fertilizer increased.Compared with the 3 g per pot level of nitrogen fertilizer and the control,the internal structure of root applied with nitrogen fertilizer of 6 g per pot level had larger fibrovascularcylinder,advanced vascular bundle,small-spaced thin-walled cells,smooth and full cells with smooth cell wall.[Conclusion]Appropriate nitrogen levels can accelerate the accumulation of soluble sugar and the growth of root tissue structure.展开更多
Soluble organic nitrogen （SON） is recognized as a sensitive indicator of soil nitrogen status. The present work was conducted in the temperate forests of northeast China where soils are typically characterized by hi...Soluble organic nitrogen （SON） is recognized as a sensitive indicator of soil nitrogen status. The present work was conducted in the temperate forests of northeast China where soils are typically characterized by high organic matter and high organic nitrogen content, and soil sampling was made in early spring just after the freeze-thaw period. The water extracted SON pools in the organic layer of forest soils were measured within the range from 156.0 mg·kg^-1 to 292.6 mg·kg^-1, a similar magnitude of salt solution extracted SON pools reported in literatures. However, the water soluble SON pools in 0-15 cm mineral soils in present study were much higher （3-10 times） than any other reports, ranging from 58.6 mg·kg^-1 to 125.2 mg·kg^-1. Water soluble SON varied markedly among the soils under different forests and at different sites. The SON in water extracts were positively and significantly correlated to soil organic matter and total nitrogen contents, but negatively correlated to microbial biomass nitrogen （MBN）. The reasons of the abnormally large SON pools and the negative correlations between SON and MBN in the 0-15cm mineral soils in this study were specially discussed.展开更多
Hydrophobically associating water-soluble polymers (HPAP) have been synthesized from acrylamide(AM), acrylate (AA), 2-acrylamido-2-methyl propane sulfonic acid (AMPS) and hydrophobic monomer (AP) in aqueous solution...Hydrophobically associating water-soluble polymers (HPAP) have been synthesized from acrylamide(AM), acrylate (AA), 2-acrylamido-2-methyl propane sulfonic acid (AMPS) and hydrophobic monomer (AP) in aqueous solution by radical polymerization. New polymer drilling fluids are made up of HPAP, which is used as viscosifiers and encapsulation agents. Properties of this system are reported in this paper. Results indicate that this system has a high value of yield point to plastic viscosity (YP/PV≥0.7), high viscosity at a low-shear rate (LSRV≥30000mPa·s), excellent shear thinning behavior, good solid-carrying behavior, resistance to shear, good thermal stability (as high as 140℃) and salt resistance. The system has excellent behavior in high-density solution of NaCl and in calcium and magnesium rich saline solutions. Hence, HPAP also can be used in saltwater polymer drilling fluids.展开更多
基金supported by the National Natural Science Foundation of China(81870259,82170431,81903608,and U21A20339)the CAMS Innovation Fund for Medical Sciences(CIFMS+1 种基金2019-I2M-5-078)the Postdoctoral Research Foundation of Heilongjiang Province(LBH-Q20148).
文摘Myocardial ischemia is a serious threat to human health,and vascular dysfunction is its main cause.Buxu Tongyu(BXTY)Granule is an effective traditional Chinese medicine(TCM)for treating myocardial ischemia.However,the underlying mechanism of BXTY is still unclear.In this study,we demonstrate that BXTY ameliorates myocardial ischemia by activating the soluble guanylate cyclase(sGC)-30,50-cyclic guanosine monophosphate(cGMP)-protein kinase G(PKG)signaling pathway in vascular smooth muscle cells(VSMCs)to dilate the arteries.BXTY was given by gavage for ten consecutive days before establishing an animal model of acute myocardial ischemia in mice via the intraperitoneal injection of pituitrin.The results showed that BXTY alleviated the symptoms of myocardial ischemia induced by pituitrin in mice,including electrocardiogram abnormalities and changes in plasma enzymes.In addition,BXTY dilated pre-constricted blood vessels and inhibited the vasoconstriction of the superior mesenteric artery in a dose-dependent but endothelial-independent manner.These effects were eliminated by preincubating vascular rings with the sGC inhibitors NS 2028 or ODQ,or with the PKG inhibitor KT 5823.Moreover,BXTY increased the protein expression of sGC-b1 and the intracellular second messenger cGMP level in mouse aortic vascular smooth muscle cells(MOVAs).NS 2028 or ODQ reversed these effects of BXTY.The expression level of the cGMP downstream effector protein PKG-1 increased after treating MOVAs with BXTY.NS 2028,ODQ,or KT 5823 also reversed this effect of BXTY.In conclusion,BXTY can improve the symptoms of acute myocardial ischemia in mice,and activating the sGC-cGMP-PKG pathway in VSMCs to induce vasodilation is its key pharmacodynamic mechanism.
文摘The increasing incidence of cardiovascular disease(CVD)is a significant global health concern,affecting millions of individuals each year.Accurate diagnosis of acute CVD poses a formidable challenge,as misdiagnosis can significantly decrease patient survival rates.Traditional biomarkers have played a vital role in the diagnosis and prognosis of CVDs,but they can be influenced by various factors,such as age,sex,and renal function.Soluble ST2(sST2)is a novel biomarker that is closely associated with different CVDs.Its low reference change value makes it suitable for continuous measurement,unaffected by age,kidney function,and other confounding factors,facilitating risk stratification of CVDs.Furthermore,the combination of sST2 with other biomarkers can enhance diagnostic accuracy and prognostic value.This review aims to provide a comprehensive overview of sST2,focusing on its diagnostic and prognostic value as a myocardial marker for different types of CVDs and discussing the current limitations of sST2.
基金financial support from National Natural Science Foundation of China (22288102)the Fundamental Research Funds for the Central Universities of China (buctrc202016)。
文摘Solubility enhancement has been a priority to overcome poor solubility with optoelectronic molecules for solution-processable devices. This study aims to obtain experimental data on the effect of particle sizes on the solubility properties of several typical optoelectronic molecules in organic solvents, including the solubility results of 1,3-bis(9-carbazolyl)benzene(m CP), 1,3,5-tris(1-phenyl-1H-benzimidazol-2-yl)ben zene(TPBi) and 2-(4-tert-butylphenyl)-5-(4-biphenyl)-1,3,4-oxadiazole(PBD) in ethanol and acetonitrile,respectively. Nanoparticles of m CP, TPBi and PBD with sizes from dozens to several hundred nanometers were prepared by solvent antisolvent precipitation method and their solubility were determined by using isothermal saturation method. The saturation solubility of nanoparticles of three kinds of optoelectronic molecules exhibited increase of 12.9%-25.7% in comparison to the same raw materials in the form of microparticles. The experimental evidence indicates that nanonization technology is a feasible way to make optoelectronic molecules dissolve in liquids with enhanced solubility.
基金supported by NIEHS(RIVER Award,R35 ES030443)NIEHS(Superfund Award,P42 ES004699)+6 种基金NINDS(Counter ActProgram U54 NS127758)Juvenile Diabetes Research Foundation(2-SRA-2022-1210-S-B)Guangzhou Science and Technology Foundation(Grant No.:201903010034)Natural Resources Science Foundation of Guangdong Province(Grant No.:2018A030313926)Science and Technology Foundation Key R&D Program of Guangdong Province(Grant Nos.:2019B020209009 and 2019B020218009)R&D Program of Guangdong Province Drug Administration(Grant Nos.:2021TDZ09 and 2021YDZ06)supported by China Scholarship Council(CSC)(202108440382).
文摘To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chromatography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.Attributes of nanobody based pharmaceutical assays are discussed.
基金Nanjing Science and Technology Plan Project(No.ZX20200009)Jiangsu Province Postgraduate Research and Practice Innovation Program(No.SJCX22-0895)。
文摘Objective:To construct a secretory eukaryotic expression vector of DSG2 fused with the Fc region of the human IgG,to validate its expression in 293T cells,and to purify the secretory protein with biological activity.Methods:The DSG2 extracellular domain fragment gene(DSG2ex),was amplified by PCR,and was inserted into the eukaryotic expression plasmid pCMV3-IgG1 to construct the recombinant eukaryotic expression plasmid-pCMV3-DSG2ex-IgG1.The successfully constructed eukaryotic expression plasmid was transfected into 293T cells to express and secrete DSG2 extracellular domain protein.The targeted protein was purified from the cell culture supernatant by Protein A affinity chromatography and confirmed by Western Blotting and ELISA.Results:The pCMV3-DSG2ex-IgG1 eukaryotic expression plasmid was successfully constructed.The highest protein expression level was obtained with 293T cells after 96 h of transfection.The relative molecular mass of the purified product was between 100 and 130 kDa was estimated by SDS-PAGE,which was consistent with the expectation.The yield of the purified protein reached 0.8 mg/ml with a purity over 90%.The purified DSG2 extracellular domain protein with IgG1 tag was recognized by IgG monoclonal antibodies by Western blotting.Moreover,the ELISA results showed that the prepared DSG2 extracellular domain protein had significant binding activity to human type 55 adenovirus Fiber Knob protein(HAdV-55).Conclusion:A simple and efficient method for eukaryotic expression and purification of human soluble DSG2 extracellular domain protein was successfully established,and biologically active DSG2 extracellular domain protein was purified,which laid the foundation for the later study of its protein function and anti-adenovirus drugs.
文摘BACKGROUND Several studies of spontaneous intracerebral hemorrhage(SICH)patients have shown apoptotic changes in brain samples after hematoma evacuation.However,there have been no data on the association between blood concentrations of soluble fas(sFas)(the main surface death receptor of the extrinsic apoptosis pathway)and the prognosis of spontaneous intracranial hypotension(SIH)patients.AIM To determine whether there is an association between blood sFas concentrations and SICH patient mortality.METHODS We included patients with severe and supratentorial SIH.Severe was defined as having Glasgow Coma Scale<9.We determined serum sFas concentrations at the time of severe SICH diagnosis.RESULTS We found that non-surviving patients(n=36)compared to surviving patients(n=39)had higher ICH score(P=0.001),higher midline shift(P=0.004),higher serum sFas concentrations(P<0.001),and lower rate of early hematoma evacuation(P=0.04).Multiple logistic regression analysis showed an association between serum sFas concentrations and 30-d mortality(odds ratio=1.070;95%confidence interval=1.014-1.129;P=0.01)controlling for ICH score,midline shift,and early hematoma evacuation.CONCLUSION The association of blood sFas concentrations and SICH patient mortality is a novel finding in our study.
文摘Bai-Hua-She-She-Cao Hedyotis diffusa Willd. (Ru-biaceae) is a medicinal herbwidely distributed in northeast Asian countries. In traditional Chinese medicine, it has the effectof 'clearing away heat and toxic material, promoting blood circulation and removing blood stasis'.It is a well known Chinese folk-medicine used for the treatment of appendicitis, sore throat, mumps,acne, sebo-rheic dermatitis and various kinds of tumors, such as tumors of digestive tract,carcinoma of liver. It was reported that the MeOH extract of H. diffusa demonstrated a significantantitumor activity and ursolic acid succeeded in being isolated from the MeOH extract as an activecomponent . Shan BN, et al suggested that the direct aqueous extract of H. diffusa hadimmuno-modulating activity and antitumor activity in vitro through stimulating the immune system tokill or engulf tumor cells. But regarding anti-tumor activity in vivo of water soluble extracts fromH. diffusa, no detail was reported. Therefore, we prepared water soluble extracts (H_1 and H_2)from H. diffusa and evaluated their anti-tumor property in vivo experiments as well as protectiveeffect on chemo-therapeutic damage.
文摘Populus tomentosa seedlings used for cold acclimation at -3℃ were pretreated with or without 10?mmol·L -1 CaCl 2, 3?mmol·L -1 of Ca 2+ chelator EGTA, 0 1?mmol·L -1 of Ca 2+ channel inhibitor LaCl 3,and 0 05?mmol·L -1 of CaM antagonist CPZ. The changes in the contents of total soluble protein and CaM, and freezing resistance in all pretreated seedlings in various periods ( viz: following cold acclimation, chilling stress and recovery) were investigated. Results showed that cold acclimation increased the contents of total soluble protein and CaM, and freezing resistance of seedlings, which could be strongly reduced by the pretreatments of EGTA CPZ and LaCl 3 Cold acclimation combined with CaCl 2 pretreatment enhanced the effect of cold acclimation on freezing resistance, and obviously increased the contents of total soluble protein and CaM, reduced the declining degree of the contents of total soluble protein and CaM caused by chilling stress as compared with cold acclimation, augmented the increase in the level of total soluble protein and CaM during the recovery periods. Further analysis found that an increase in total soluble protein content during cold acclimation with or without CaCl 2 pretreatment mainly resulted from the increase in content of heat stable protein in total soluble protein. It is suggested that Ca 2+ calmodulin may be involved in the synthesis of total soluble protein, and the induction of freezing resistance of seedlings.
文摘The changes in the contents of total soluble protein and RNA, the activity of RNase in leaves and branches of Populus tomentosa cuttings at various periods (viz: cold acclimation, deacclimation, chilling stress and the recovery after chilling stress), and the survival rate and the freezing resistance of cuttings during cold acclimation at -3℃ were investigated. Results showed that cold acclimation not only increased the contents of total soluble protein and RNA, the survival rates and the freezing resistance of cuttings, decreased the activity of RNase, but also reduced the declining degree of total soluble protein and RNA contents, and the increasing level of RNase caused by chilling stress as compared with the controls. In addition, cold acclimation augmented the increase in the level of total soluble protein and RNA, and facilitated the decrease of RNase during the recovery periods. Further analysis found that the DNA content of all treatments kept relative stability at various periods. The changes in total soluble protein, RNA and RNase were closely related to the freezing resistance of cuttings. It appears that the increase of RNA content caused by cold acclimation induced decrease of RNase activity may be involved in the accumulation of total soluble protein and the induction of freezing resistance of cuttings.
基金NationalNaturalScienceFoundationofChina (No .59772 0 1 9)
文摘Pervoskite type oxides LaCoO 3 was prepared by citrate method with the granula of 20 nm-30 nm. With a fluorescent Hg lamp or sunlight as irradiator, the degradation experiments of various water soluble dyes were carried out in the suspension system of LaCoO 3 . The results show that the pervoskite type oxide LaCoO 3 has good photocatalytic activity.Studied by X ray photoelectron spectroscopy and photoacoustic spectra, its photocatalytic activity is found to be related with factors such as the d electron structure of ion Co 3+ ,Co—O binding energy and adsorbed oxygen on the surface etc.
基金Supported by the National Natural Science Foundation of China (40901115,31070548,31170578)the Foundation of Fujian Province (2009J01207)~~
文摘[Objective] The extraction efficiency of soluble nitrogen in common moun- tain red soil under different conditions was analyzed to provide experimental refer- ences for the extraction of soil soluble nitrogen. [Method] The fresh soil and air-dried soil, which were the common mountain red soil under evergreen broad-leaved forest vegetation, were used as the experimental materials. The 2 mol/L KCI and 0.5 mol/L K2SO4 were used as the extractants to extract the soil soluble nitrogen with the extractant-to-soil volume ratio at 4：1, 6：1 and 10：1, respectively, to examine the extrac- tion efficiency of soluble nitrogen in the experimental soil with different extraction conditions. [Result] Compared with K2SO4, the NH4＋-N extracted by KCI was better in quantity and stability; while only at high extractant-to-soil volume ratio could K2SO4 fully extract the soil NH4＋-N. The 2 extractants showed small difference in extracting NO3--N, but at low extractant-to-soil volume ratio, the stability of NO3--N extracted by KCI was higher than that extracted by K2SO4. The properties of NO2- -N extracted by the 2 extractants were very similar. As for soil SON extraction, there was small dif- ference between the 2 extractants. However, the SON extracted by KCI was fewer in quantity than that by K2SO4, but had higher stability, and the extracted SON amount of the 2 extractants showed significant correlation. The extractant-to-soil ratio had little effect on MBN extraction using K2SO4, but the effect was significant when using KCI to extract MBN. The difference between the extraction efficiency of KCI and K^SO4 in extracting fresh soils was greater than that in extracting air-dried soil; therefore, the extractants should be carefully selected in soluble nitrogen extraction from fresh soil. [Conclusion] The results lay the experimental foundation for improving the extraction efficiency of soil soluble nitrogen and increasing the comparability of measurement results.
基金supported by the National Natural Science Foundation of China (No. 40706050, 40706048 and 30700619)the National Science & Technology Pillar Program (No. 2006BAD01A13 and 2008BAC49B04)+2 种基金National special fund for transgenic project (No. 2009ZX08009-019B)Natural Science Foundation of Shandong Province (No. 2009ZRA02075)and Qingdao Municipal Science and Technology plan project (No. 09-2-5-8-hy)
文摘Total soluble proteins of different life stages, filamentous sporophytes cultivated in high temperatures, and blade gametophytes harvested in different seasons, were identified by SDS-PAGE. The types and amounts of expressed proteins also varied amongst the samples. The fewest soluble proteins were present in filamentous sporophytes. There were more types and amounts of soluble protein in conchospores than in filamentous sporophytes, but fewer than in bulgy sporophytes. More types of protein were detected in filamentous sporophytes cultivated in high temperatures than in those growing in normal situations. The most types and amounts of protein were found in blade gametophytes in all samples. Blade gametophytes harvested last year and stored at -20 ℃ showed only minor differences in expression of proteins when compared with those harvested in different seasons.
文摘[Objective] The research aimed to provide theoretical basis for the cultivation and production of peach.[Method] The three-year-old seedling peach tree was used as the materials,and NH4NO3 was used as the experimental nitrogen fertilizer.Three nitrogen levels,0,3,6 g per pot respectively were set to study the effects of different nitrogen fertilizer on the contents of soluble sugar,starch and internal structure of the root of the peaches.[Result] The contents of soluble sugar of roots and leaves increased with the increase of nitrogen fertilizer level,and the level of 6 g per pot was the highest,which showed extreme differences between the level of 3 g per pot and the control.However,starch contents showed opposite results,which decreased as the level of nitrogen fertilizer increased.Compared with the 3 g per pot level of nitrogen fertilizer and the control,the internal structure of root applied with nitrogen fertilizer of 6 g per pot level had larger fibrovascularcylinder,advanced vascular bundle,small-spaced thin-walled cells,smooth and full cells with smooth cell wall.[Conclusion]Appropriate nitrogen levels can accelerate the accumulation of soluble sugar and the growth of root tissue structure.
基金National Nature Science Foundation of China (30571476, 30771703)
文摘Soluble organic nitrogen （SON） is recognized as a sensitive indicator of soil nitrogen status. The present work was conducted in the temperate forests of northeast China where soils are typically characterized by high organic matter and high organic nitrogen content, and soil sampling was made in early spring just after the freeze-thaw period. The water extracted SON pools in the organic layer of forest soils were measured within the range from 156.0 mg·kg^-1 to 292.6 mg·kg^-1, a similar magnitude of salt solution extracted SON pools reported in literatures. However, the water soluble SON pools in 0-15 cm mineral soils in present study were much higher （3-10 times） than any other reports, ranging from 58.6 mg·kg^-1 to 125.2 mg·kg^-1. Water soluble SON varied markedly among the soils under different forests and at different sites. The SON in water extracts were positively and significantly correlated to soil organic matter and total nitrogen contents, but negatively correlated to microbial biomass nitrogen （MBN）. The reasons of the abnormally large SON pools and the negative correlations between SON and MBN in the 0-15cm mineral soils in this study were specially discussed.
文摘Hydrophobically associating water-soluble polymers (HPAP) have been synthesized from acrylamide(AM), acrylate (AA), 2-acrylamido-2-methyl propane sulfonic acid (AMPS) and hydrophobic monomer (AP) in aqueous solution by radical polymerization. New polymer drilling fluids are made up of HPAP, which is used as viscosifiers and encapsulation agents. Properties of this system are reported in this paper. Results indicate that this system has a high value of yield point to plastic viscosity (YP/PV≥0.7), high viscosity at a low-shear rate (LSRV≥30000mPa·s), excellent shear thinning behavior, good solid-carrying behavior, resistance to shear, good thermal stability (as high as 140℃) and salt resistance. The system has excellent behavior in high-density solution of NaCl and in calcium and magnesium rich saline solutions. Hence, HPAP also can be used in saltwater polymer drilling fluids.
