Objective The aim of this study was to characterize the basic molecular and biochemical parameters for a cyclophilin protein in Cryptosporidium parvum called CpCyP1.Methods CpCyP1 expression patterns during the parasite life cycle were evaluated using qRT-PCR with total RNA isolated from different developmental stages of C.parvum.Native CpCyP1 protein in sporozoites was detected using western blot.The localization of CpCyP1 was performed using the immunofluorescence assay,with an affinity-purified rabbit polyclonal antibody against a synthetic peptide.The peptidyl-prolyl cis-trans isomerase(PPIase)activity of His-tagged recombinant CpCyP1 was evaluated using absorbance colorimetry,and the effect of cyclosporin A(CsA)on the activity of CpCyP1 was determined.Results CpCyP1 was expressed in all parasite developmental stages,whereas CpCyP1 was present mainly in the cytosol of sporozoites,meronts,and gamonts.CpCyP1 displayed Michaelis-Menten kinetics towards N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide for its PPIase activity(Km=456.4 μmol/L;Vmax=1.981 U).CsA inhibited PPIase activity,showing lower micromolar inhibitory activity and binding affinity(Kd=5.122 μmol/L;IC50=1.004 μmol/L).Conclusions These results imply that CpCyP1 in the parasite may be the target for the previously reported anti-cryptosporidial efficacy of CsA and suggest that C.parvum cyclophilins could be evaluated as candidate drug targets.

Objective To explore the situation of impaired cardiac microcirculatory perfusion,left ventricular remodeling and myocardial fibrosis in patients with essential hypertension(abbreviated as hypertension)and their clinical significance.Methods Eighty patients with hypertension were selected as hypertension group,and another 80 healthy individuals who underwent physical examina-tions during the same period were selected as control group.Based on the severity of hypertension,patients in the hypertension group were divided into low-risk group and moderate-to-high-risk group.The indicators of impaired cardiac microcirculatory perfusion[left ventricular myocardial blood flow(MBF),right ventricular MBF,coronary flowreserve(CFR)],left ventricular remodeling[left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV)]and myocardial fibrosis[left ventricular isovolumic relaxation time(IVRT),the ratio of the peak velocity of E wave to A wave in the diastolic blood flow spectrum of the mitral valve orifice(E/A),type Ⅰ procollagen carboxy-terminal peptide(P Ⅰ CP),type Ⅲprocollagen amino-terminal peptide(PC Ⅲ)]were compared between the control group and the hy-pertension group.Pearson's correlation coefficient method was used for correlation analysis.The differences in the above-mentioned indicators among different risk groups were compared.Receiver operating characteristic(ROC)curves were drawn to evaluate the diagnostic efficacy of impaired cardiac microcirculatory perfusion,left ventricular remodeling and myocardial fibrosis for the severity of hypertension.Results In the hypertension group,left ventricular MBF,right ventricular MBF and CFR were all lower than those in the control group,LVEDV and LVESV were larger than those in the control group,LVEF was lower than that in the control group,IVRT was longer than that in the control group,the levels of P Ⅰ CP and PC Ⅲ were higher than those in the control group,and E/A was lower than that in the control group(P＜0.05).The results of Pearson's correlation coef-ficient analysis showed that LVEF was positively correlated with CFR and E/A,and negatively cor-related with IVRT and PC Ⅲ;LVEDV was positively correlated with CFR,IVRT,P Ⅰ CP and PCⅢ;LVESV was positively correlated with IVRT,P Ⅰ CP and PC Ⅲ,and negatively correlated with E/A;left ventricular MBF was positively correlated with CFR and E/A,and negatively correlated with IVRT,P Ⅰ CP and PC Ⅲ;right ventricular MBF was positively correlated with CFR and E/A,and negatively correlated with IVRT,P Ⅰ CP and PC Ⅲ.In the moderate-to-high-risk group,IVRT was longer than that in the low-risk group,LVEDV was larger than that in the low-risk group,P ⅠCP,PC Ⅲ,E/A,LVEF,left ventricular MBF,right ventricular MBF and CFR were all lower than those in the low-risk group,and LVESV was smaller than that in the low-risk group,with statistical-ly significant differences(P＜0.05).The results of ROC curve analysis showed that the areas under the curves for diagnosing the severity of hypertension by impaired cardiac microcirculatory perfusion,left ventricular remodeling and myocardial fibrosis were 0.962,0.969 and 0.945 respectively,the sensitivities were 91.4％,97.1％and 88.6％respectively,and the specificities were 95.6％,88.9％and 93.3％respectively.Conclusion Hypertension can cause impaired cardiac microcirculatory perfusion and promote the processes of left ventricular remodeling and myocardial fibrosis.Impaired cardiac microcirculatory perfusion,left ventricular remodeling and myocardial fibrosis have good di-agnostic efficacy for the severity of hypertension.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

BACKGROUND:In recent years,functional near-infrared spectroscopy has shown extensive research significance and application potential in the fields of cognitive neuroscience,clinical medicine,and engineering technology due to its portability,low cost,non-invasiveness,and high temporal resolution.OBJECTIVE:To analyze the research progress,hot issues,and frontier trends of functional near-infrared spectroscopy in the past 10 years.METHODS:Web of Science database was searched for articles related to functional near-infrared spectroscopy published from January 1,2013 to November 30,2023.CiteSpace and VOSviewer software was used to conduct a visual analysis of publication statistics,journals,countries,institutions,authors,and keywords.RESULTS AND CONCLUSION:This article included a total of 2 714 documents,showing an overall upward trend in the volume of literature concerning functional near-infrared spectroscopy over the past decade.The 2 714 documents selected for this article were authored by 9 171 researchers from 2 400 institutions across 68 countries in 495 kinds of journals.The United States and University College London are representative countries and institutions,while Ehlis and Fallgatter are two influential authors in this research field.The journal NeuroImage ranks first with the highest citation rate.Functional near-infrared spectroscopy research primarily comes from University College London in the UK,Beijing Normal University in China,and the University of Tübingen in Germany.The keyword co-occurrence network map generated using VOSviewer 1.6.20 reveals six research clusters.By analyzing the specific keywords of each cluster,we further understand the research hotspots in this field.The burst analysis of keywords uncovers the research frontiers and development trends,providing important guidance for future research directions of functional near-infrared spectroscopy.

BACKGROUND:In recent years,functional near-infrared spectroscopy has shown extensive research significance and application potential in the fields of cognitive neuroscience,clinical medicine,and engineering technology due to its portability,low cost,non-invasiveness,and high temporal resolution.OBJECTIVE:To analyze the research progress,hot issues,and frontier trends of functional near-infrared spectroscopy in the past 10 years.METHODS:Web of Science database was searched for articles related to functional near-infrared spectroscopy published from January 1,2013 to November 30,2023.CiteSpace and VOSviewer software was used to conduct a visual analysis of publication statistics,journals,countries,institutions,authors,and keywords.RESULTS AND CONCLUSION:This article included a total of 2 714 documents,showing an overall upward trend in the volume of literature concerning functional near-infrared spectroscopy over the past decade.The 2 714 documents selected for this article were authored by 9 171 researchers from 2 400 institutions across 68 countries in 495 kinds of journals.The United States and University College London are representative countries and institutions,while Ehlis and Fallgatter are two influential authors in this research field.The journal NeuroImage ranks first with the highest citation rate.Functional near-infrared spectroscopy research primarily comes from University College London in the UK,Beijing Normal University in China,and the University of Tübingen in Germany.The keyword co-occurrence network map generated using VOSviewer 1.6.20 reveals six research clusters.By analyzing the specific keywords of each cluster,we further understand the research hotspots in this field.The burst analysis of keywords uncovers the research frontiers and development trends,providing important guidance for future research directions of functional near-infrared spectroscopy.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

Objective To evaluate the angiogenesis of decidua in spontaneous abortion patients and to investigate the change of thyroid hormone receptor signal expression in decidua in miscarriage patients.Methods The decidua of pregnant women was collected.Patients with miscarriage were taken as the experimental group and healthy preg-nant women who underwent abortion by social factors were taken as the control group.The nuclear translocation of thyroid hormone receptor α(THR-α)and THR-β in decidua tissues was detected by Western blot method,and the distribution of THR-α and TH IR-β in decidua tissues was determined by immunohistochemistry.The expression of Deiodinase 2(DIO2)in decidua tissues was determined by Western blot.The distribution of DIO2 in decidua tissues was determined by immunofluorescence and the fluorescence intensity was measured.The decidual vessel density was determined by immunohistochemistry using CD34 as an endothelial marker.The protein expression of vascular endothelial growth factor receptor(VEGFR-1)was detected by Western blot.Results There was no sig-nificant difference in age,gestation days,TSH,T3,T4 and other basic data between the control group and the a-bortion group.The results of Western blot showed that the expression levels of THR-α and THR-β protein de-creased,and the results of immunohistochemical staining of decidual tissues showed that the nuclear translocation of decidual thyroid hormone receptor in patients with spontaneous abortion decreased(P＜0.05).Western blotting results showed that the expression of DIO2 protein in the abortion group was lower than that in the control group,suggesting that thyroid hormone metabolism in decidua of spontaneous abortion pregnant women was affected.The results of immunofluorescence test showed that the fluorescence intensity of DIO2 in the abortion group was weaker than that in the control group(P＜0.05).Immunohistochemical CD34 labeling of decidual small blood vessels showed that the number of CD34+blood vessels decreased in the abortion group with statistical difference(P＜0.05).Western blot results showed that there was no significant change in VEGFR-1 between the two groups,and the difference between the two groups was not statistically significant.Conclusion The abnormal signal of thyroid hormone receptors in decidua may decrease decidua angiogenesis and participate in spontaneous abortion.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.

An electrochemical chemical oxygen demand(COD)sensor was proposed based on a FTO/TiO2/PbO2 electrode and a thin-layer electrochemical cell.The FTO/TiO2/PbO2 electrode was characterized by X-ray photoelectronic spectroscopy(XPS),X-ray diffraction(XRD)spectroscopy and electrochemical technique,and the results indicated that the rapid decrease in the output signals of the electrochemical COD sensor could be attributed to oxidation of PbSO4 occurring on the surface of FTO/TiO2/PbO2 electrode.The PbO2 deposition time and concentration of Na2SO4 were further optimized and then the electrochemical COD sensor was challenged by real samples including laker water sample,river water sample and wastewater sample.The evolution trend of signals of the electrochemical COD sensor in response to lake and river water samples was identical with that obtained with the standard method(HJ/T399-2007,Water quality-determination of the chemical oxygen demand-fast digestion-spectrophotometric method).The electrochemical COD sensor exhibited significant increase in the signal intensity after the samples were switched from lake water to wastewater sample,and a mean value of 32.5 mg/L with relative standard deviation(RSD)of 6.8%were obtained after measuring 45 times the wastewater with COD value of 30 mg/L under a sampling interval of 400 s.The as-prepared electrochemical COD sensor possessed good promise in regular monitoring of COD,discharge of wastewater and industrial process control,with advantages such as a small sampling interval,mild reaction conditions and no requirement of toxic and harmful chemical reagents.

Objective To verify the safety and efficacy of a new portable extracorporeal membrane oxygenation(ECMO)system(Xijing Advanced Life Support System JC-Ⅲ)in large animals.Methods A total of 10 healthy small fat-tail sheep underwent veno-arterial extracorporeal membrane oxygenation(VA-ECMO)support by carotid arterial-jugular catheterization to evaluate the performance of the JC-Ⅲ ECMO system.Systemic anticoagulation was achieved by continuous infusion of heparin.Active coagulation time(ACT)was recorded every 2 hours during the experiment,and the ACT was maintained between 200-250 s.Centrifugal pump speed is set at 3 000-3 500 r/min.The changes of hemoglobin,blood cell counts,hematocrit,liver and kidney function were monitored before and 24 h after ECMO initiation,respectively.After the experiment,the pump and oxygenator were dissected to probe the thrombosis.Results The success rate of VA-ECMO operation was 100％,and there was no hemolysis,pump thrombosis and oxygenator thrombosis after 24 h of ECMO.Before and after the operation,there were no significant changes in indicators such as hemoglobin content,white blood cell counts,platelet counts,alanine aminotransferase concentration,aspartate aminotransferase concentration,urea,creatinine,high-sensitivity troponin Ⅰ,and N-terminal pro-brain natriuretic peptide(all P＞0.05).Conclusions This in vivo study confirms that Xijing Advanced Life support System JC-Ⅲ is safe and effective.