Objective:To analyze and predict the biological properties and function of Treponema pallidum OmpH protein by bioinformatics methods, purify the target protein, and prepare polyclonal antibodies. Methods:From January 2024 to February 2025, the research team from the Department of Clinical Laboratory at The First Affiliated Hospital of Hunan Traditional Chinese Medical College (Hunan Province Directly Affiliated Traditional Chinese Medical Hospital) conducted a study employing integrative approaches combining bioinformatics analysis with animal experimentation. During this investigation, the coding sequence of the T. pallidum outer membrane protein H (TpOmpH) was systematically retrieved from the National Center for Biotechnology Information (NCBI) database. And the bioinformatics tools, such as Protparam, Protscale,SignalP 6.0,NetNGlyc-1.0,TMHMM2.0,NetPhos-3.1,SOPMA,AlphaFold3,IEDB,STRING,C-immsim were used to analyze and predict the biological and immunological characteristics of TpOmpH protein. The full length of TpOmpH gene was synthesized and was cloned into the pET28a to construct the recombinant plasmid pET28a-TpOmpH. The the expression of target protein was induced by IPTG and was purified using affinity chromatography. The TpOmpH protein was used to immunize mice and the anti-serum was harvested, then the titer of antibody was detected. Results:TpOmpH is a hydrophobic outer membrane protein with a molecular weight of 19.7 kDa and strong stability. The TpOmpH protein is located outside the cell membrane and contains 11 serine, 4 threonine, and 1 tyrosine phosphorylation site, but no glycosylation sites. The 77.91% of the amino acids in TpOmpH protein are alpha helix, 8.72% are extended strand, 10.47% are random coils, and 2.91% are beta turns. The tertiary structure predicted by AlphaFold3 is in its optimal state. The TpOmpH protein has 4 CTL epitopes, 4 linear epitopes, and 5 spatial epitopes. The TpOmpH protein can interact with Tp92,MutS,SurA,TPANIC_0600 and other proteins which may be involved in Tp invasion. TpOmpH protein can induce an increase in B cell count, antibody content, Th cell count, NK cell count, as well as the expression of various cytokines. High purity TpOmpH protein was obtained through Ni 2+ affinity chromatography, which is consistent with the theoretical molecular weight. TpOmpH protein can induce mice to secrete polyclonal antibodies with antibody titers higher than 1∶10 000. Conclusion:TpOmpH protein is a hydrophobic protein located on the outer membrane of Tp, can induce mice to secrete high titer antibodies, which providing experimental basis for the pathogenesis of Tp and vaccine development.

AIM To study the chemical constituents from salt-processed Litchi Semen and their antioxidant activities.METHODS The 85％ethanol extract from salt-processed Litchi Semen was isolated and purified by silica gel,Sephadex LH-20,MCI,ODS and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.DPPH and ABTS+free radical scavenging method were used to evaluate their antioxidant activities.RESULTS Fifteen compounds were isolated and identified as dehydrocostuslactone(1),ananosmoside A(2),funingensin A(3),(2S)-pinocembrin-7-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranoside)(4),liquiritienin(5),quercetin(6),rutin(7),isorhamnetin-3-O-β-rutinoside(8),procyanidin A2(9),procyanidin A1(10),ethyl protocatechuate(11),5-hydroxymethylfurfural(12),di(2-ethyl-hexyl)phthalate(13),nicotinamide(14),(10E,15Z)-9,12,13-trihydroxyoctadeca-10,15-dienoic acid(15).Compounds 6-7,9-10 exhibited scavenging activities against DPPH radicals with IC50 values of(12.929±1.232),(14.104±0.946),(10.417±1.736),(6.944±0.030)μmol/L,respectively.Compounds 6-10 exhibited scavenging activities against ABTS+radicals with IC50 values of(21.952±0.577),(25.683±0.625),(22.970±1.336),(20.210±1.435),(18.725±0.324)μmol/L,respectively.CONCLUSION Compounds 1,5,14-15 are isolated from Litchi genus for the first time.Compounds 6-7,9-10 have strong in vitro antioxidant activities.

Objective To evaluate the effects of transcatheter aortic valve replacement(TAVR)on left ventricular reverse remodeling(LVRR)and outcomes in patients with mixed aortic valve disease(MAVD)and predominant aortic stenosis(AS).Methods Patients undergoing TAVR at our center between January 2020 and December 2022 were enrolled consecutively.Propensity score matching(PSM)(1∶1 ratio)was used to reduce selection bias.Transthoracic echocardiography(TTE)was used to monitor left ventricular ejection fraction(LVEF)and other structural parameters over time.The study outcome was a composite of cardiovascular death and rehospitalization due to cardiovascular causes.Linear mixed-effects models and logistic regression were utilized for comparing echocardiographic changes across groups and identifying independent risk factors for no-LVRR,respectively.Results After PSM,126 patients were included.MAVD group exhibited larger structural parameters(left ventricular end-systolic/end-diastolic diameter and volume,left ventricular mass index)and a lower left ventricular ejection fraction(LVEF)(all P＜0.05).However,more pronounced improvements in left ventricular structure and hemodynamics were observed during follow-up.Multivariate logistic regression analysis indicated that the left ventricular mass index(LVMI)was an independent predictor of left ventricular reverse remodeling(LVRR)after TAVR,whereas persistent moderate or greater mitral regurgitation(MR)and paravalvular leak(PVL)significantly reduced the incidence of LVRR.During a median follow-up period of 23 months,a total of 31 endpoint events occurred,and there was no statistically significant difference in long-term prognosis between the two groups(Log-rank P=0.330).Conclusions Compared to patients in the AS group,those in the MAVD group exhibited more severe left ventricular remodeling before TAVR.However,more significant LVRR was observed during postoperative follow-up.Additionally,the long-term prognosis was comparable between the two groups.

AIM To study the chemical constituents from salt-processed Litchi Semen and their antioxidant activities.METHODS The 85％ethanol extract from salt-processed Litchi Semen was isolated and purified by silica gel,Sephadex LH-20,MCI,ODS and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.DPPH and ABTS+free radical scavenging method were used to evaluate their antioxidant activities.RESULTS Fifteen compounds were isolated and identified as dehydrocostuslactone(1),ananosmoside A(2),funingensin A(3),(2S)-pinocembrin-7-O-(6-O-α-L-rhamnopyranosyl-β-D-glucopyranoside)(4),liquiritienin(5),quercetin(6),rutin(7),isorhamnetin-3-O-β-rutinoside(8),procyanidin A2(9),procyanidin A1(10),ethyl protocatechuate(11),5-hydroxymethylfurfural(12),di(2-ethyl-hexyl)phthalate(13),nicotinamide(14),(10E,15Z)-9,12,13-trihydroxyoctadeca-10,15-dienoic acid(15).Compounds 6-7,9-10 exhibited scavenging activities against DPPH radicals with IC50 values of(12.929±1.232),(14.104±0.946),(10.417±1.736),(6.944±0.030)μmol/L,respectively.Compounds 6-10 exhibited scavenging activities against ABTS+radicals with IC50 values of(21.952±0.577),(25.683±0.625),(22.970±1.336),(20.210±1.435),(18.725±0.324)μmol/L,respectively.CONCLUSION Compounds 1,5,14-15 are isolated from Litchi genus for the first time.Compounds 6-7,9-10 have strong in vitro antioxidant activities.

Metabolic associated fatty liver disease(MAFLD)has a complex pathogenesis,and mitophagy is involved in the development and progression of MAFLD and plays a key role in liver metabolic pathways and signaling networks.Mitophagy is regulated by a variety of pathways,and the PTEN-induced kinase 1(PINK1)/Parkin pathway is considered the main pathway for regulating mitophagy.Mitophagy mediated by the PINK1/Parkin pathway can regulate lipid metabolism,inflammation,and fibrosis and delay the progression of MAFLD.This article reviews the role of mitophagy mediated by the PINK1/Parkin pathway in MAFLD and the research advances in targeted therapy,in order to provide theoretical bases and ideas for the prevention and treatment of MAFLD.

Aim To evaluate the role of the glucose transporter protein 1(GLUT1)-dependent glycolytic in the attenuation of oxygen-glucose deprivation-reoxygen-ation(OGD/R)injury in HK-2 cells by dexmedetomi-dine(Dex).Methods C57/BL6 mice were random-ly divided into three groups(n=6),namely,sham operation group(Sham group),renal ischemia reper-fusion group(I/R group)and Dex group(I/R+Dex group).Serum creatinine(Cr)and urea nitrogen(BUN)were measured,while the levels of key glyco-lytic enzymes HK2,PFKFB3 and GLUT1 were meas-ured.HK-2 cells were cultured and randomised into seven groups(n=6),which was treated with OGD/R,overexpression or interference with GLUT1,Dex and glycolysis inhibitor 2-DG.CCK-8 and LDH activi-ty were used to detect cellular damage.Glycolysis lev-els were detected by lactate and ECAR.The inflamma-tory level was reflected by qRT-PCR for IL-6 and TNF-α.qRT-PCR and Western blot were performed to de-tect the levels of GLUT1,HK2,and PFKFB3.Results Dex significantly ameliorated kidney injury and HK-2 cell injury(P＜0.05).Dex inhibited the OGD/R-induced rise in lactate and extracellular acidification rate(ECAR),as evidenced by suppression of the ex-pression of GLUT1,HK2 and PFKFB3(P＜0.05).In vitro experiments showed that GLUT1 knockdown sig-nificantly improved OGD/R-induced cellular damage.Lactate,ECAR,glycolysis-related mRNAs and pro-teins were inhibited by GLUT1 knockdown(P＜0.05).Significantly,there were no significant differ-ences in above indexes after Dex treatment based on GLUT1 knockdown.Overexpression of GLUT1 abroga-ted the protective effects of Dex,while reversing the inhibitory effects of Dex on the expression of GLUT1,HK2,and PFKFB3(P＜0.05).Conclusions Dexmedetomidine attenuates OGD/R induced injury in HK-2 cells by inhibiting GLUT1-dependent glycolysis.

Objective:To analyze and predict the biological properties and function of Treponema pallidum OmpH protein by bioinformatics methods, purify the target protein, and prepare polyclonal antibodies. Methods:From January 2024 to February 2025, the research team from the Department of Clinical Laboratory at The First Affiliated Hospital of Hunan Traditional Chinese Medical College (Hunan Province Directly Affiliated Traditional Chinese Medical Hospital) conducted a study employing integrative approaches combining bioinformatics analysis with animal experimentation. During this investigation, the coding sequence of the T. pallidum outer membrane protein H (TpOmpH) was systematically retrieved from the National Center for Biotechnology Information (NCBI) database. And the bioinformatics tools, such as Protparam, Protscale,SignalP 6.0,NetNGlyc-1.0,TMHMM2.0,NetPhos-3.1,SOPMA,AlphaFold3,IEDB,STRING,C-immsim were used to analyze and predict the biological and immunological characteristics of TpOmpH protein. The full length of TpOmpH gene was synthesized and was cloned into the pET28a to construct the recombinant plasmid pET28a-TpOmpH. The the expression of target protein was induced by IPTG and was purified using affinity chromatography. The TpOmpH protein was used to immunize mice and the anti-serum was harvested, then the titer of antibody was detected. Results:TpOmpH is a hydrophobic outer membrane protein with a molecular weight of 19.7 kDa and strong stability. The TpOmpH protein is located outside the cell membrane and contains 11 serine, 4 threonine, and 1 tyrosine phosphorylation site, but no glycosylation sites. The 77.91% of the amino acids in TpOmpH protein are alpha helix, 8.72% are extended strand, 10.47% are random coils, and 2.91% are beta turns. The tertiary structure predicted by AlphaFold3 is in its optimal state. The TpOmpH protein has 4 CTL epitopes, 4 linear epitopes, and 5 spatial epitopes. The TpOmpH protein can interact with Tp92,MutS,SurA,TPANIC_0600 and other proteins which may be involved in Tp invasion. TpOmpH protein can induce an increase in B cell count, antibody content, Th cell count, NK cell count, as well as the expression of various cytokines. High purity TpOmpH protein was obtained through Ni 2+ affinity chromatography, which is consistent with the theoretical molecular weight. TpOmpH protein can induce mice to secrete polyclonal antibodies with antibody titers higher than 1∶10 000. Conclusion:TpOmpH protein is a hydrophobic protein located on the outer membrane of Tp, can induce mice to secrete high titer antibodies, which providing experimental basis for the pathogenesis of Tp and vaccine development.

The 16S rRNA sequencing,whole genome sequencing and drug sensitivity tests were used to identify the isolates molecularly and to detect and analyse their virulence genes,resistance genes and drug resistance.The results showed that the isolate was highly homologous to Klebsiella pneumoniae X4 and located on the same branch by 16S rRNA sequence analysis,and it was named as KLKp10.Whole genome sequencing results showed that the KLKp10 genome was 5 342 841 bp in length,containing 5 138 genes,346 repetitive segments,6 rRNAs and 81 tRNAs,with a GC con-tent of 57.30％.MLST analysis showed that KLKp10 belongs to the ST-4263 type.The functions of 4 097 of the genes encoding proteins were classified and annotated by COG,and there were also 382 genes with unknown functions.A total of 50 functional classifications were involved in the an-notation results based on the GO database;33 kinds of signaling pathways were covered based on the signaling pathway annotations in the KEGG database.A total of 443 virulence genes were screened in the VFDB database,of which 339 belonged to the Set A database and could encode 124 virulence factors.The 101 resistance genes were predicted by comparing with the CARD database,among which there were more resistance genes against β-lactam antibiotics.The results of drug sensitivity test showed that KLKp10 was highly sensitive to ceftazidime,gentamicin,azithro-mycin,chloramphenicol,norfloxacin,ofloxacin,and enrofloxacin;moderately sensitive to ceftriax-one,neomycin,kanamycin,and streptomycin;and resistant to ciprofloxacin,tetracycline,amoxicil-lin,and penicillin.In this study,we systematically revealed the gene-wide characterization,virulence factors and drug resistance of Klebsiella pneumoniae KLKp10 of Kole pig origin,which provides important data support for the study of Klebsiella pneumoniae at the overall level of its genome.

The 16S rRNA sequencing,whole genome sequencing and drug sensitivity tests were used to identify the isolates molecularly and to detect and analyse their virulence genes,resistance genes and drug resistance.The results showed that the isolate was highly homologous to Klebsiella pneumoniae X4 and located on the same branch by 16S rRNA sequence analysis,and it was named as KLKp10.Whole genome sequencing results showed that the KLKp10 genome was 5 342 841 bp in length,containing 5 138 genes,346 repetitive segments,6 rRNAs and 81 tRNAs,with a GC con-tent of 57.30％.MLST analysis showed that KLKp10 belongs to the ST-4263 type.The functions of 4 097 of the genes encoding proteins were classified and annotated by COG,and there were also 382 genes with unknown functions.A total of 50 functional classifications were involved in the an-notation results based on the GO database;33 kinds of signaling pathways were covered based on the signaling pathway annotations in the KEGG database.A total of 443 virulence genes were screened in the VFDB database,of which 339 belonged to the Set A database and could encode 124 virulence factors.The 101 resistance genes were predicted by comparing with the CARD database,among which there were more resistance genes against β-lactam antibiotics.The results of drug sensitivity test showed that KLKp10 was highly sensitive to ceftazidime,gentamicin,azithro-mycin,chloramphenicol,norfloxacin,ofloxacin,and enrofloxacin;moderately sensitive to ceftriax-one,neomycin,kanamycin,and streptomycin;and resistant to ciprofloxacin,tetracycline,amoxicil-lin,and penicillin.In this study,we systematically revealed the gene-wide characterization,virulence factors and drug resistance of Klebsiella pneumoniae KLKp10 of Kole pig origin,which provides important data support for the study of Klebsiella pneumoniae at the overall level of its genome.

Background:The incidence and mortality rates of severe acute pancreatitis(SAP)have been increasing year by year.Therefore,early and rapid identification,along with timely intervention in the progression of acute pancreatitis(AP),is of particular importance.Aims:To explore the value of red blood cell distribution width(RDW)combined with BISAP score in the early prediction of SAP.Methods:A total of 561 AP patients admitted from January 2019 to December 2021 at the General Hospital of the Central Theater Command of the PLA were enrolled and divided into SAP group and non-SAP group according to the disease severity.Venous blood samples were collected within 24 hours of admission.The relevant clinical data,laboratory indices,BISAP score,and MCTSI score were compared between the two groups.Logistic regression analysis was used to identify the risk factors for SAP.Spearman correlation coefficient was employed to assess the correlation of these risk factors with the severity of AP,as well as the correlation of RDW with BISAP score and MCTSI score.The predictive values of these risk factors for SAP were evaluated by ROC curve analysis.Results:Compared with the non-SAP group,the prevalence of hypertension,length and cost of hospital stay,neutrophil count(NEUT),neutrophil-to-lymphocyte ratio(NLR),RDW,serum potassium,aspartate transaminase(AST),blood urea nitrogen(BUN),serum creatinine(SCr),BISAP score and MCTSI score were significantly increased in the SAP group(all P<0.05),while the lymphocyte count(LYM),serum calcium and albumin(ALB)were significantly decreased(all P<0.05).RDW(OR=1.582,95%CI:1.066-2.348,P=0.023),SCr(OR=1.018,95%CI:1.001-1.035,P=0.040),BISAP score(OR=6.210,95%CI:3.121-12.356,P<0.001),and MCTSI score(OR=2.917,95%CI:2.160-3.939,P<0.001)were the independent risk factors for SAP.RDW(rs=0.320,P<0.001),SCr(rs=0.103,P=0.015),BISAP score(rs=0.516,P<0.001),and MCTSI score(rs=0.512,P<0.001)were positively correlated with the severity of AP.Moreover,RDW was positively correlated with BISAP score(rs=0.428,P<0.001)and MCTSI score(rs=0.408,P<0.001).ROC curve analysis revealed that the areas under the ROC curve of RDW,SCr,BISAP score,MCTSI score,and combination of RDW and BISAP score for predicting SAP were 0.753,0.581,0.889,0.888,and 0.905,respectively.Conclusions:RDW,SCr,BISAP score,and MCTSI score are the independent risk factors for SAP.RDW combined with BISAP score can enhance the predictive value for SAP.