Abstract Thermal inactivation of a commercial β-galactosidase from Aspergillus oryzae in a 300 g/L lactose solution was studied in the temperature range of 65–75 °C. Lactose exhibited a stabilisation effect when similar inactivation rates as those in lactose solution were observed in a lactose-free solution at temperatures lower by 5°C. Inactivation process in the lactose solution was biphasic. A kinetic model based on the Lumry-Eyring mechanism was proposed and successfully verified. Estimated activation energy values were very different. Rather high activation energy values of the forward reactions were responsible for both the significant change of rate constants and the rate-controlling reaction with temperature. For these two reasons, an increase of the operational lifetime of the enzyme from 7 days at 60 °C to 580 days at 55 °C was predicted.
{"title":"Thermal inactivation kinetics of Aspergillus oryzae β-galactosidase in concentrated lactose solution","authors":"V. Illeová, M. Polakovič","doi":"10.2478/acs-2018-0024","DOIUrl":"https://doi.org/10.2478/acs-2018-0024","url":null,"abstract":"Abstract Thermal inactivation of a commercial β-galactosidase from Aspergillus oryzae in a 300 g/L lactose solution was studied in the temperature range of 65–75 °C. Lactose exhibited a stabilisation effect when similar inactivation rates as those in lactose solution were observed in a lactose-free solution at temperatures lower by 5°C. Inactivation process in the lactose solution was biphasic. A kinetic model based on the Lumry-Eyring mechanism was proposed and successfully verified. Estimated activation energy values were very different. Rather high activation energy values of the forward reactions were responsible for both the significant change of rate constants and the rate-controlling reaction with temperature. For these two reasons, an increase of the operational lifetime of the enzyme from 7 days at 60 °C to 580 days at 55 °C was predicted.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"170 - 174"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46856738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Panthenol is a biologically active compound closely related to vitamin B5 (pantothenic acid). This work deals with the separation of panthenol enantiomers using high performance liquid chromatography. Different types of chiral stationary phases (β-cyclodextrin, isopropyl carbamate cyclofructan 6, amylose tris(3,5-dimethylphenylcarbamate)) were tested in normal phase separation mode. Effect of mobile phase composition on the resolution and retention of enantiomers was studied. Two types of detectors, low-wavelength UV and polarimetric, were used. The optimal chromatographic system includes a chiral stationary phase based on amylose and a mobile phase of hexane/ethanol (60/40, v/v) where the resolution of enantiomers reached the value Rs = 2.49. Suitable chromatographic conditions were applied for the determination of panthenol enantiomers in samples of pharmaceutical preparations with the obtained recovery of more than 92 %. Linearity of the high performance liquid chromatography method with spectrophotometric detection was from 1.0 × 10−3 to 1.3 mg mL−1 (R2 = 0.998), with the limit of detection of 0.3 × 10−3 mg mL−1 for both enantiomers.
{"title":"HPLC separation of panthenol enantiomers on different types of chiral stationary phases","authors":"Anna Lomenova, K. Hroboňová, Terézia Šolónyová","doi":"10.2478/acs-2018-0017","DOIUrl":"https://doi.org/10.2478/acs-2018-0017","url":null,"abstract":"Abstract Panthenol is a biologically active compound closely related to vitamin B5 (pantothenic acid). This work deals with the separation of panthenol enantiomers using high performance liquid chromatography. Different types of chiral stationary phases (β-cyclodextrin, isopropyl carbamate cyclofructan 6, amylose tris(3,5-dimethylphenylcarbamate)) were tested in normal phase separation mode. Effect of mobile phase composition on the resolution and retention of enantiomers was studied. Two types of detectors, low-wavelength UV and polarimetric, were used. The optimal chromatographic system includes a chiral stationary phase based on amylose and a mobile phase of hexane/ethanol (60/40, v/v) where the resolution of enantiomers reached the value Rs = 2.49. Suitable chromatographic conditions were applied for the determination of panthenol enantiomers in samples of pharmaceutical preparations with the obtained recovery of more than 92 %. Linearity of the high performance liquid chromatography method with spectrophotometric detection was from 1.0 × 10−3 to 1.3 mg mL−1 (R2 = 0.998), with the limit of detection of 0.3 × 10−3 mg mL−1 for both enantiomers.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"114 - 119"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42192602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Marcela Múdra, M. Breza, Lucia Lintnerová, J. Filo, J. Bauer
Abstract A strategy for the design of new yttrium(III) tags consisting of sequences of naturally occurring amino acids is described. These tags are 4–6 amino acids in length and consist of aspartic and glutamic acids. The use of natural amino acids would allow these oligopeptides to be incorporated into recombinant proteins at the DNA level, enabling the protein to be Y(III)-labelled after protein isolation. This allows a radionuclide or heavy atom to be associated with the protein without the necessity of further synthetic modification. Suitable peptides able to chelate Y(III) in stable complexes were designed based on quantum-chemical calculations. The stability of complexes formed by these peptides was tested by isothermal titration calorimetry, giving dissociation constants in the micromolar range. The likely structure of the most tightly bound complex was inferred from a combination of NMR experiments and quantum-chemical calculations. This structure will serve as the basis for future optimizations.
{"title":"The design and NMR structure determination of yttrium-oligopeptide tags for recombinant proteins and antibodies","authors":"Marcela Múdra, M. Breza, Lucia Lintnerová, J. Filo, J. Bauer","doi":"10.2478/acs-2018-0018","DOIUrl":"https://doi.org/10.2478/acs-2018-0018","url":null,"abstract":"Abstract A strategy for the design of new yttrium(III) tags consisting of sequences of naturally occurring amino acids is described. These tags are 4–6 amino acids in length and consist of aspartic and glutamic acids. The use of natural amino acids would allow these oligopeptides to be incorporated into recombinant proteins at the DNA level, enabling the protein to be Y(III)-labelled after protein isolation. This allows a radionuclide or heavy atom to be associated with the protein without the necessity of further synthetic modification. Suitable peptides able to chelate Y(III) in stable complexes were designed based on quantum-chemical calculations. The stability of complexes formed by these peptides was tested by isothermal titration calorimetry, giving dissociation constants in the micromolar range. The likely structure of the most tightly bound complex was inferred from a combination of NMR experiments and quantum-chemical calculations. This structure will serve as the basis for future optimizations.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"120 - 133"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2478/acs-2018-0018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44305038","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract The title compound, C14H20INO, is a molecule with three stereogenic centres. It absolute configuration was derived from the synthesis and confirmed by structure determination (AD, Flack (Parsons’) parameter: 0.031 (8)). The expected stereochemistry of atoms N1 was confirmed to be S, C5 was confirmed to S, C6 was confirmed to R. The central N-heterocyclic ring is not planar and adopts a half-chair conformation. A calculation of least-squares planes showed that these rings are puckered in such a manner that the five atoms: C5, C6, C7, C12 and C13 (the second ring: C1, C2, C3, C4, C5 and N1) are planar, while atom N1 is displaced from these plane with the out-of-plane displacement of −0.694 (4) and −0.670 (5) Å in the second ring, respectively. Dihedral angle between the planes of the central N-heterocyclic rings is 23.4 (2)°. Crystal structure is also stabilized by C—H···O hydrogen interactions.
{"title":"Crystal, molecular and electronic structure of (5S,11R,11aS)-11-hydroxy-5-methyl-1,2,3,4,5,6,11,11a-octahydropyrido[1,2-b]isoquinolin-5-ium iodide","authors":"J. Sivý, P. Šafár̆, J. Žúžiová","doi":"10.2478/acs-2018-0014","DOIUrl":"https://doi.org/10.2478/acs-2018-0014","url":null,"abstract":"Abstract The title compound, C14H20INO, is a molecule with three stereogenic centres. It absolute configuration was derived from the synthesis and confirmed by structure determination (AD, Flack (Parsons’) parameter: 0.031 (8)). The expected stereochemistry of atoms N1 was confirmed to be S, C5 was confirmed to S, C6 was confirmed to R. The central N-heterocyclic ring is not planar and adopts a half-chair conformation. A calculation of least-squares planes showed that these rings are puckered in such a manner that the five atoms: C5, C6, C7, C12 and C13 (the second ring: C1, C2, C3, C4, C5 and N1) are planar, while atom N1 is displaced from these plane with the out-of-plane displacement of −0.694 (4) and −0.670 (5) Å in the second ring, respectively. Dihedral angle between the planes of the central N-heterocyclic rings is 23.4 (2)°. Crystal structure is also stabilized by C—H···O hydrogen interactions.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"94 - 98"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46714197","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Arsenic (As) is metalloid, naturally present in the environment but also introduced by human activities. It is toxic and carcinogenic and its exposure to low or high concentrations can be fatal to human health. Arsenic contamination in drinking water threatens more than 150 million peoples all over the world. Therefore, treatment of As contaminated water is of unquestionable importance. The present review begins with an overview of As chemistry, distribution and toxicity, which are relevant aspects to understand and develop remediation techniques. The most common As removal processes (chemical precipitation, adsorption, ion exchange, membrane filtration, phytoremediation and electrocoagulation) are presented with discussion of their advantages, drawbacks and the main recent achievements.
{"title":"An overview of main arsenic removal technologies","authors":"Ronald Zakhar, J. Derco, F. Cacho","doi":"10.2478/acs-2018-0016","DOIUrl":"https://doi.org/10.2478/acs-2018-0016","url":null,"abstract":"Abstract Arsenic (As) is metalloid, naturally present in the environment but also introduced by human activities. It is toxic and carcinogenic and its exposure to low or high concentrations can be fatal to human health. Arsenic contamination in drinking water threatens more than 150 million peoples all over the world. Therefore, treatment of As contaminated water is of unquestionable importance. The present review begins with an overview of As chemistry, distribution and toxicity, which are relevant aspects to understand and develop remediation techniques. The most common As removal processes (chemical precipitation, adsorption, ion exchange, membrane filtration, phytoremediation and electrocoagulation) are presented with discussion of their advantages, drawbacks and the main recent achievements.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"107 - 113"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47822583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract A summary of the preparation methods of 2 tautomeric and 4 N-methylated benzimidazoles with a nitro group on the benzene ring (1–6) and with an amino group in the same positions (7–12) were summarized. Annular tautomerism of the title compounds 1–12 has been studied using 1H, 13C and 15N NMR spectra in liquid and solid state (CPMAS), UV spectra and quantum chemical calculations.
{"title":"Nitro and aminobenzimidazoles","authors":"V. Milata","doi":"10.2478/acs-2018-0026","DOIUrl":"https://doi.org/10.2478/acs-2018-0026","url":null,"abstract":"Abstract A summary of the preparation methods of 2 tautomeric and 4 N-methylated benzimidazoles with a nitro group on the benzene ring (1–6) and with an amino group in the same positions (7–12) were summarized. Annular tautomerism of the title compounds 1–12 has been studied using 1H, 13C and 15N NMR spectra in liquid and solid state (CPMAS), UV spectra and quantum chemical calculations.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"182 - 188"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2478/acs-2018-0026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48096399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract This paper deals with the analysis and design of a model predictive control (MPC) strategy used in connection with level control in conically shaped industrial liquid storage tanks. The MPC is based on a non-linear dynamic model describing changes of the liquid level concerning changes in the inlet flow of the liquid. Euler discretization of the dynamic system was applied to transform con-tinuous time dynamics to its discrete-time counterpart used in non-linear MPC (NMPC) design. By means of a simulation case study, NMPC has been shown to track the changes of the liquid level, hence provides increased control performance. This paper also compares the traditional approach of optimal control, linear MPC, with the NMPC strategy.
{"title":"Non-linear model predictive control of conically shaped liquid storage tanks","authors":"Martin Klauco, Ľ. Čirka","doi":"10.2478/acs-2018-0020","DOIUrl":"https://doi.org/10.2478/acs-2018-0020","url":null,"abstract":"Abstract This paper deals with the analysis and design of a model predictive control (MPC) strategy used in connection with level control in conically shaped industrial liquid storage tanks. The MPC is based on a non-linear dynamic model describing changes of the liquid level concerning changes in the inlet flow of the liquid. Euler discretization of the dynamic system was applied to transform con-tinuous time dynamics to its discrete-time counterpart used in non-linear MPC (NMPC) design. By means of a simulation case study, NMPC has been shown to track the changes of the liquid level, hence provides increased control performance. This paper also compares the traditional approach of optimal control, linear MPC, with the NMPC strategy.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"141 - 146"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47000220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
D. Cagardová, M. Michalík, Peter Poliak, Vladimír Lukes
Abstract A systematic theoretical study using density functional theory is presented to estimate the structural, electronic, and charge-transfer characteristics of a symmetric fluorination of acenequinones outer rings. The change in aromaticity of model derivatives was described by different types of aromaticity indices. By considering a hopping mechanism and using the Marcus theory in combination with the Einstein-Smoluchowski relation, electronic drift mobilities were predicted for selected dimer configurations obtained from X-ray structures of anthraquinone, 6,13-pentacenequinone and its octafluorinated derivatives. The analysis of obtained data showed that the fluorination of the outer rings of acenequinones can lower the energy of the lowest unoccupied molecular orbital to the range from −3.0 to −4.0 eV, i.e. typical for organic n-type semiconducting materials. Finally, potential electric semiconductivity of available X-ray structures relating to drift mobilities was discussed.
{"title":"Quantum chemical study of electron structure and charge transport properties of symmetric acenequinones","authors":"D. Cagardová, M. Michalík, Peter Poliak, Vladimír Lukes","doi":"10.2478/acs-2018-0013","DOIUrl":"https://doi.org/10.2478/acs-2018-0013","url":null,"abstract":"Abstract A systematic theoretical study using density functional theory is presented to estimate the structural, electronic, and charge-transfer characteristics of a symmetric fluorination of acenequinones outer rings. The change in aromaticity of model derivatives was described by different types of aromaticity indices. By considering a hopping mechanism and using the Marcus theory in combination with the Einstein-Smoluchowski relation, electronic drift mobilities were predicted for selected dimer configurations obtained from X-ray structures of anthraquinone, 6,13-pentacenequinone and its octafluorinated derivatives. The analysis of obtained data showed that the fluorination of the outer rings of acenequinones can lower the energy of the lowest unoccupied molecular orbital to the range from −3.0 to −4.0 eV, i.e. typical for organic n-type semiconducting materials. Finally, potential electric semiconductivity of available X-ray structures relating to drift mobilities was discussed.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"83 - 93"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48251845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Mališová, M. Horňáček, J. Mikulec, P. Hudec, V. Jorík
Abstract The aim of the research was the study of aluminium-magnesium hydrotalcite by FTIR analysis and detection of differences in structure due to calcination. Three types of samples were analysed. In the first two samples, cobalt was added into the structure and the third sample contained nickel and zinc. Hydrotalcite was prepared by one of the most common methods of hydrotalcite preparation, co-precipitation. After preparation, the samples were calcined at temperatures from 150 °C to 750 °C. Samples were compared in terms of calcination temperature; but in case of samples containing cobalt, two samples to each other. The most significant changes were noticed in the 500–700 cm−1 region, where a spinel structure was formed at higher calcination temperatures.
{"title":"FTIR study of hydrotalcite","authors":"M. Mališová, M. Horňáček, J. Mikulec, P. Hudec, V. Jorík","doi":"10.2478/acs-2018-0021","DOIUrl":"https://doi.org/10.2478/acs-2018-0021","url":null,"abstract":"Abstract The aim of the research was the study of aluminium-magnesium hydrotalcite by FTIR analysis and detection of differences in structure due to calcination. Three types of samples were analysed. In the first two samples, cobalt was added into the structure and the third sample contained nickel and zinc. Hydrotalcite was prepared by one of the most common methods of hydrotalcite preparation, co-precipitation. After preparation, the samples were calcined at temperatures from 150 °C to 750 °C. Samples were compared in terms of calcination temperature; but in case of samples containing cobalt, two samples to each other. The most significant changes were noticed in the 500–700 cm−1 region, where a spinel structure was formed at higher calcination temperatures.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"147 - 156"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43758469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Thermal inactivation of immobilized glucose isomerase in a concentrated glucose solution was investigated in the batch mode and temperature range of 83–95 °C, which is substantially higher than the temperature used in the industrial production of high-fructose corn syrup. Simultaneous evaluation of all inactivation data showed that first-order kinetics with the Arrhenius temperature dependence of the rate constant provided a good approximation of the biocatalyst stability under the investigated conditions. The model parameters were then used to predict the operational temperature for this biocatalyst in the production of high-fructose corn syrup based on the set operational life-time of the biocatalyst. The simulation predicted a window of operational temperature of 60–65 °C, which corresponds very well with the industrial applications of this biocatalyst. This observation demonstrates that the multi-temperature method of enzyme inactivation can provide a good estimate of biocatalyst process stability and is thus a useful tool in the development of biocatalytic processes.
{"title":"Design of operational temperature for immobilized glucose isomerise using an accelerated inactivation method","authors":"V. Illeová, M. Polakovič","doi":"10.2478/acs-2018-0022","DOIUrl":"https://doi.org/10.2478/acs-2018-0022","url":null,"abstract":"Abstract Thermal inactivation of immobilized glucose isomerase in a concentrated glucose solution was investigated in the batch mode and temperature range of 83–95 °C, which is substantially higher than the temperature used in the industrial production of high-fructose corn syrup. Simultaneous evaluation of all inactivation data showed that first-order kinetics with the Arrhenius temperature dependence of the rate constant provided a good approximation of the biocatalyst stability under the investigated conditions. The model parameters were then used to predict the operational temperature for this biocatalyst in the production of high-fructose corn syrup based on the set operational life-time of the biocatalyst. The simulation predicted a window of operational temperature of 60–65 °C, which corresponds very well with the industrial applications of this biocatalyst. This observation demonstrates that the multi-temperature method of enzyme inactivation can provide a good estimate of biocatalyst process stability and is thus a useful tool in the development of biocatalytic processes.","PeriodicalId":7088,"journal":{"name":"Acta Chimica Slovaca","volume":"11 1","pages":"157 - 162"},"PeriodicalIF":0.8,"publicationDate":"2018-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44676375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：