Objective To explore the clinical characteristics as well as molecular epidemiological features of resis-tance genes and virulence genes of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection in a region,and provide scientific basis for the prevention,treatment,and epidemiological study of CRKP.Methods 60 non-repeti-tive CRKP strains isolated clinically from Puyang Oilfield General Hospital from November 2023 to September 2024 were analyzed retrospectively.Antimicrobial susceptibility testing was performed using VITEK 2 Compact automa-tic microbial analyzer,K-B disk diffusion method,and micro-broth dilution method.Mucus phenotype of bacterial strains was identified by string test.Carbapenemase was detected by carbapenemase inhibitor enhancement assay.Molecular features,such as multi-locus sequence typing(MLST),capsule serotypes,resistance genes,virulence genes,plasmid replication types of strains,as well as the genetic and evolutionary relationships of strains were de-termined by whole genome sequencing and bioinformatics analysis.Results CRKP strains were mainly isolated from elderly male hospitalized patients.Specimens were mostly from sputum(71.67％),mainly distributed in depart-ment of respiratory medicine(30.00％).All strains were highly resistant to multiple commonly used antimicrobial agents,only with high susceptibility rates to cefotaxime/avibactam,tigecycline,and polymyxin B(＞60.00％).Two CRKP strains were positive for string test.95.00％of the strains produced class A serine carbapenemase.All strains carried fluoroquinolone,phosphomycin,β-lactam,and aminoglycoside resistance genes;enterobactin,Esche-richia coli common pilus(ECP),and outer membrane protein-related virulence genes;as well as plasmids from the IncF plasmid family.Carbapenemase gene was mainly blaKPC-2(95.00％),and the major capsule serotype was KL19(43.33％).In MLST,ST11(51.67％)was the dominant clone group,and ST11-KL62(n=12)was the dominant subtype.Conclusion CRKP in this hospital is highly resistant to multiple commonly used antimicrobial agents,and its mechanism of resistance to carbapenems is mainly related to the presence of blaKPC-2 resistance gene.All strains have coexisting multiple resistance genes and virulence genes,and show a phenomenon of multi-clone transmission.ST11 is the dominant clone group,and ST11-KL62 is the main prevalent subclone type.

BACKGROUND:Percutaneous retrograde screws and minimally invasive percutaneous plates are extensively used in the treatment of unilateral pubic ramus fractures.However,the above internal fixations have the disadvantages of mistakenly penetrating the hip joint and damaging the medial pelvic trophoblastic artery.A new type of retrograde superior pubic ramus intramedullary nail offers advantages,such as fewer radiographic exposures and shorter surgery times,but the validation of the biomechanical properties of the three endoprostheses is lacking.OBJECTIVE:To compare the biomechanical differences post-treatment of upper and lower pubic ramus fractures using percutaneous retrograde screws,minimally invasive percutaneous plates,and retrograde superior pubic ramus intramedullary nail under various conditions through finite element analysis.METHODS:Pelvic CT data were collected from one volunteer and imported into Mimics 20,Geomagic Wrap 2021,and SolidWorks 2019 to create geometric models of three types of internal fixation for treating unilateral pubic ramus fractures:hollow screw group,plate group,and intramedullary nail group.Each model was imported into Ansys 2022 software,where it was set up in vertical,book-opening,and shear conditions for preprocessing and calculation to compare and analyze the mechanical stability of the implants.RESULTS AND CONCLUSION:(1)In vertical conditions,the biomechanical stability of the hollow screw treatment for unilateral pubic fractures was superior to that of the steel plate and intramedullary nail,with the smallest peak stress in the implant,which was four times lower compared to the other two groups.(2)In book-opening conditions,the steel plate treatment for unilateral pubic fractures showed better biomechanical stability,particularly in reducing stress at the fracture ends of the upper pubic branch,with stress values five times higher in the other two groups.(3)In shear conditions,the intramedullary nail treatment for unilateral pubic fractures exhibited good biomechanical stability,maintaining the best pelvic displacement,with a 25％greater displacement observed in the other two groups.(4)These findings reveal the biomechanical differences of different implants:the peak stress of implants is the smallest under vertical working conditions with percutaneous retrograde screws.Minimally invasive percutaneous plates reduce the stress at the broken end of superior pubic branch under open-book working condition.Retrograde superior pubic ramus intramedullary nail of superior pubic branch maintains pelvic displacement under shear working condition.The surgeon can combine the biomechanical differences of the implants with the actual situation of pubic injury in clinical practice to make the best implant treatment.

AIM:To investigate the role of BRCA2 and CDKN1A interacting protein(BCCIP)in gastric can-cer(GC)and elucidate its mechanism in mediating cisplatin resistance.METHODS:The BCCIP mRNA expression was assessed in GC tissues(n=415)and normal tissues(n=34)using The Cancer Genome Atlas(TCGA)database.In an in-ternal cohort(n=36 for RT-qPCR;n=5 for Western blot;n=30 for immunohistochemistry),BCCIP expression at both mRNA and protein levels was examined in GC tissues and paired adjacent normal tissues.Human GC cell lines AGS and HGC27 were cultured in vitro and treated with cisplatin in a dose(0,2,4,6,8 and 10 μmol/L)-and time(0,6,24 and 48 h)-dependent manner,followed by Western blot analysis of BCCIP expression.Stable BCCIP knockdown cell lines(shRNA#1 and shRNA#2 groups)were generated via lentiviral transfection,with empty vector-transfected cells serving as controls(vector group).Flow cytometry and colony formation assay were performed to evaluate the effects of BCCIP on apoptosis and colony-forming ability of GC cells treated with cisplatin.Western blot was utilized to detect the changes of BCCIP protein expression levels in the cytoplasm and nucleus of GC cells after cisplatin(2.5 and 1.0 μmol/L)treatment,as well as the effects of BCCIP on the expression of DNA damage marker γ-H2AX and apoptosis-related proteins cleaved caspase-9 and cleaved caspase-3,and the activation of checkpoint kinase 1(CHK1)after cisplatin(2.5 and 1.0 μmol/L)treatment.Immunofluorescence was conducted to observe the effect of BCCIP on γ-H2AX expression in GC cells treated with cisplatin(2.5 and 1.0 μmol/L).RESULTS:The BCCIP expression was significantly up-regulated in GC tissues compared with normal tissues(P<0.01).Cisplatin induced up-regulation of BCCIP expression in a dose-and time-depen-dent manner.Knockdown of BCCIP significantly enhanced cisplatin-induced apoptosis(P<0.01)and reduced colony-forming ability(P<0.05)of GC cells.Knockdown of BCCIP promoted the expression of γ-H2AX,but inhibited the activa-tion of CHK1 after cisplatin treatment,with increased protein levels of cleaved caspase-9 and cleaved caspase-3(P<0.01).CONCLUSION:Cisplatin promotes the expression of BCCIP in GC cells.BCCIP confers cisplatin resistance in GC cells by suppressing apoptosis through modulation of DNA damage response pathways.

Aim To study the diastolic effect and mechanism of farrerol on isolated pulmonary arteries of C57BL/6J mice.Methods After anesthesia,mouse lung tissue was quickly removed and placed into the 4 ℃ K-H buffer,pulmonary arteries were isolated under the microscope and cut into 2 mm long vascular rings for spare use.(1)The effect of farrerol on the resting tension of isolated mouse pulmonary arteries:in the resting state,the active mouse pul-monary artery rings were treated with different concentrations of farrerol(10-6,3×10-6,10-5,3×10-5 and 10-4 mol/L).(2)Farrerol relaxed mouse pulmonary artery experiment:pulmonary arteries were contracted using phenylephrine(PE,1 μmol/L)or KCl(60 mmol/L),and when the contraction reached the platform,different concentrations of farrerol(10-6,3×10-6,10-5,3×10-5 and 10-4 mol/L)was added.(3)Farrerol inhibited pulmonary artery contraction experi-ment:under conditions with or without the addition of farrerol,pulmonary arteries were contracted using different concen-trations of PE(10-9,3×10-9,10-8,3 × 10-8,10-7,3×10-7 and 10-6 mol/L)or KCl(20,30,40,60,80 and 120 mmol/L),and the pulmonary artery muscle tension was recorded.(4)Calcium free and recalcification experiments:under conditions with or without the addition of farrerol,the changes of isolated mouse pulmonary artery tension were meas-ured in the state of calcium free or recalcification { 2.5 mmol/L[Ca2+]ex }.(5)The relationship between farrerol in-duced relaxation of isolated mouse pulmonary arteries and potassium ion channels:firstly,60 mmol/L KCl solution was used to contract the mouse pulmonary arteries until the platform.Then,3 mmol/L aminopyridine(4-AP),2 mmol/L tet-raethylammonium(TEA),30 μmol/L BaCl2,and 10 μmol/L glibenclamide(Gli)were added and treated for 15 min.Subsequently,the pulmonary arteries were relaxed using a concentration gradient of farrerol.Results Farrerol had no significant effect on the mouse pulmonary arteries in the resting state,but had a concentration-dependent relaxing effect on the mouse pulmonary arteries pre-contracted with PE and KCl.While the pretreatment of 3×10-5 mol/L farrerol could sig-nificantly reduce the maximum contraction of mouse pulmonary arteries induced by PE and KCl(P＜0.01),as well as sig-nificantly reduce the contraction of mouse pulmonary arteries induced by KCl under calcium free or recalcification conditions(P＜0.01).Addition of the voltage-dependent potassium ion channel blocker 4-AP significantly reduced the maximum diastolic rate of mouse pulmonary arteries induced by farrerol(P＜0.01),while addition of the high conductivity calcium activated potassium ion channel blocker TEA,inward rectifying potassium ion channel blocker BaCl2,or ATP sensitive po-tassium ion channel blocker Gli had no significant effect on the vasodilation effect of farrerol(P＞0.05).Conclusion Farrerol has a relaxing effect on isolated mouse pulmonary arteries,and its mechanism may be related to open voltage-de-pendent potassium ion channels.

Objective To explore the clinical characteristics as well as molecular epidemiological features of resis-tance genes and virulence genes of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection in a region,and provide scientific basis for the prevention,treatment,and epidemiological study of CRKP.Methods 60 non-repeti-tive CRKP strains isolated clinically from Puyang Oilfield General Hospital from November 2023 to September 2024 were analyzed retrospectively.Antimicrobial susceptibility testing was performed using VITEK 2 Compact automa-tic microbial analyzer,K-B disk diffusion method,and micro-broth dilution method.Mucus phenotype of bacterial strains was identified by string test.Carbapenemase was detected by carbapenemase inhibitor enhancement assay.Molecular features,such as multi-locus sequence typing(MLST),capsule serotypes,resistance genes,virulence genes,plasmid replication types of strains,as well as the genetic and evolutionary relationships of strains were de-termined by whole genome sequencing and bioinformatics analysis.Results CRKP strains were mainly isolated from elderly male hospitalized patients.Specimens were mostly from sputum(71.67％),mainly distributed in depart-ment of respiratory medicine(30.00％).All strains were highly resistant to multiple commonly used antimicrobial agents,only with high susceptibility rates to cefotaxime/avibactam,tigecycline,and polymyxin B(＞60.00％).Two CRKP strains were positive for string test.95.00％of the strains produced class A serine carbapenemase.All strains carried fluoroquinolone,phosphomycin,β-lactam,and aminoglycoside resistance genes;enterobactin,Esche-richia coli common pilus(ECP),and outer membrane protein-related virulence genes;as well as plasmids from the IncF plasmid family.Carbapenemase gene was mainly blaKPC-2(95.00％),and the major capsule serotype was KL19(43.33％).In MLST,ST11(51.67％)was the dominant clone group,and ST11-KL62(n=12)was the dominant subtype.Conclusion CRKP in this hospital is highly resistant to multiple commonly used antimicrobial agents,and its mechanism of resistance to carbapenems is mainly related to the presence of blaKPC-2 resistance gene.All strains have coexisting multiple resistance genes and virulence genes,and show a phenomenon of multi-clone transmission.ST11 is the dominant clone group,and ST11-KL62 is the main prevalent subclone type.

BACKGROUND:Percutaneous retrograde screws and minimally invasive percutaneous plates are extensively used in the treatment of unilateral pubic ramus fractures.However,the above internal fixations have the disadvantages of mistakenly penetrating the hip joint and damaging the medial pelvic trophoblastic artery.A new type of retrograde superior pubic ramus intramedullary nail offers advantages,such as fewer radiographic exposures and shorter surgery times,but the validation of the biomechanical properties of the three endoprostheses is lacking.OBJECTIVE:To compare the biomechanical differences post-treatment of upper and lower pubic ramus fractures using percutaneous retrograde screws,minimally invasive percutaneous plates,and retrograde superior pubic ramus intramedullary nail under various conditions through finite element analysis.METHODS:Pelvic CT data were collected from one volunteer and imported into Mimics 20,Geomagic Wrap 2021,and SolidWorks 2019 to create geometric models of three types of internal fixation for treating unilateral pubic ramus fractures:hollow screw group,plate group,and intramedullary nail group.Each model was imported into Ansys 2022 software,where it was set up in vertical,book-opening,and shear conditions for preprocessing and calculation to compare and analyze the mechanical stability of the implants.RESULTS AND CONCLUSION:(1)In vertical conditions,the biomechanical stability of the hollow screw treatment for unilateral pubic fractures was superior to that of the steel plate and intramedullary nail,with the smallest peak stress in the implant,which was four times lower compared to the other two groups.(2)In book-opening conditions,the steel plate treatment for unilateral pubic fractures showed better biomechanical stability,particularly in reducing stress at the fracture ends of the upper pubic branch,with stress values five times higher in the other two groups.(3)In shear conditions,the intramedullary nail treatment for unilateral pubic fractures exhibited good biomechanical stability,maintaining the best pelvic displacement,with a 25％greater displacement observed in the other two groups.(4)These findings reveal the biomechanical differences of different implants:the peak stress of implants is the smallest under vertical working conditions with percutaneous retrograde screws.Minimally invasive percutaneous plates reduce the stress at the broken end of superior pubic branch under open-book working condition.Retrograde superior pubic ramus intramedullary nail of superior pubic branch maintains pelvic displacement under shear working condition.The surgeon can combine the biomechanical differences of the implants with the actual situation of pubic injury in clinical practice to make the best implant treatment.

A 59-year-old male patient with sigmoid colon cancer and liver metastasis received the treatments of cetuximab combined with FOLFIRI regimen (irinotecan, calcium folinate, and fluorouracil). His serum creatinine (Scr) was 82 μmol/L, and estimated glomerular filtration rate (eGFR) was 96 ml/(min·1.73 m 2) before the treatment. On the 11th day after finishing the 4th cycle of treatments, the patient developed nausea and vomiting. Laboratory tests showed Scr 221 μmol/L, eGFR 29 ml/(min·1.73 m 2), serum albumin 29.0 g/L, urinary protein (+++), and 24 hour urine protein quantified 8.88 g. He was diagnosed as having nephrotic syndrome complicated with acute kidney injury, which was considered to be related to cetuximab. The drug was stopped and symptomatic treatments such as anti-inflammatory, anticoagulation and kidney protection were given. After 14 days, laboratory tests showed no proteinuria, Scr 156 μmol/L, and eGFR 44 ml/(min·1.73 m 2). Prednisone acetate tablet 50 mg were orally administered once daily. Fifty-nine days later, his Scr was 142 μmol/L, and eGFR was 49 ml/(min·1.73 m 2). Chemotherapy was suspended and cetuximab was discontinued permanently.

Aim To explore the influence of demethy-lase fat mass and obesity-related genes(FTO)on the abnormal contractile function of diabetic coronary smooth muscle.Methods Smooth muscle specific FTO knockout mice(FTOSMKO)were prepared by Cre-loxP recombinant technology.They were divided into four groups:control(WT)group,diabetes model group(DM),FTO knockout group(FTOSMKO),and FTOSMKO diabetic group(FTOSMKO-DM),with 15 mice in each group.Diabetic mice were prepared by intraperitoneal injection of streptozotocin(STZ);the remaining mice were injected with an equal amount of citric acid-sodi-um citrate buffer.The effects of 5-HTon the contractile response of coronary artery smooth muscle in the four groups of mice were observed by the technique of small-vessel ring tensiometry.Western blot and Dot blot were used to detect the changes of FTO protein and N6-methyladenine(m6A)methylation modification levels in mouse vascular tissues.Results Compared with the WT group,the DM group had significantly higher blood glucose(P＜0.01)and lower body weight(P＜0.05);the level of FTO protein in aorta of DM group increased(P＜0.01),and the level of m6A methylation modification decreased(P＜0.01).The 5-HT-induced contractile response significantly de-creased in the DM group compared with the WT group(P＜0.01),whereas the contractile response signifi-cantly increased in the FTOSMKO-DM group compared with the DM group(P＜0.01);the non-L-type calci-um channel-mediated vascular smooth muscle contrac-tile response was enhanced in the FTOSMKO-DM group,among which,the contractions induced by 1,4,5-triphosphate inositol receptor(IP3R)and caffeine-ac-tivated ranine receptor(RyR)mediated by sarcoplas-mic reticulum calcium release both significantly in-creased(P＜0.05).Conclusions Specific knock-down of smooth muscle FTO improves coronary artery responsiveness to the vasoconstrictor 5-HT in diabetic mice,which may be related to abnormalities in the FTO-mediated 5-HT receptor signaling pathway.

AIM:To investigate the role of BRCA2 and CDKN1A interacting protein(BCCIP)in gastric can-cer(GC)and elucidate its mechanism in mediating cisplatin resistance.METHODS:The BCCIP mRNA expression was assessed in GC tissues(n=415)and normal tissues(n=34)using The Cancer Genome Atlas(TCGA)database.In an in-ternal cohort(n=36 for RT-qPCR;n=5 for Western blot;n=30 for immunohistochemistry),BCCIP expression at both mRNA and protein levels was examined in GC tissues and paired adjacent normal tissues.Human GC cell lines AGS and HGC27 were cultured in vitro and treated with cisplatin in a dose(0,2,4,6,8 and 10 μmol/L)-and time(0,6,24 and 48 h)-dependent manner,followed by Western blot analysis of BCCIP expression.Stable BCCIP knockdown cell lines(shRNA#1 and shRNA#2 groups)were generated via lentiviral transfection,with empty vector-transfected cells serving as controls(vector group).Flow cytometry and colony formation assay were performed to evaluate the effects of BCCIP on apoptosis and colony-forming ability of GC cells treated with cisplatin.Western blot was utilized to detect the changes of BCCIP protein expression levels in the cytoplasm and nucleus of GC cells after cisplatin(2.5 and 1.0 μmol/L)treatment,as well as the effects of BCCIP on the expression of DNA damage marker γ-H2AX and apoptosis-related proteins cleaved caspase-9 and cleaved caspase-3,and the activation of checkpoint kinase 1(CHK1)after cisplatin(2.5 and 1.0 μmol/L)treatment.Immunofluorescence was conducted to observe the effect of BCCIP on γ-H2AX expression in GC cells treated with cisplatin(2.5 and 1.0 μmol/L).RESULTS:The BCCIP expression was significantly up-regulated in GC tissues compared with normal tissues(P<0.01).Cisplatin induced up-regulation of BCCIP expression in a dose-and time-depen-dent manner.Knockdown of BCCIP significantly enhanced cisplatin-induced apoptosis(P<0.01)and reduced colony-forming ability(P<0.05)of GC cells.Knockdown of BCCIP promoted the expression of γ-H2AX,but inhibited the activa-tion of CHK1 after cisplatin treatment,with increased protein levels of cleaved caspase-9 and cleaved caspase-3(P<0.01).CONCLUSION:Cisplatin promotes the expression of BCCIP in GC cells.BCCIP confers cisplatin resistance in GC cells by suppressing apoptosis through modulation of DNA damage response pathways.

Rice (Oryza sativa L.), as a thermophilic crop, is highly susceptible to cold stress during its growth process. Chilling injury at the plumule stage and seedling stage often affects the morphological development and leads to yield reduction of rice. The exploration and utilization of cold tolerance genes are among the most direct and effective approaches to address cold stress in rice. To identify quantitative trait loci (QTLs) associated with cold tolerance at plumule and seedling stages, in this study, we measured the seedling rates and survived seedling rates of the indica rice cultivar 'HZ', the japonica cultivar 'Nekken2', and their 120 recombinant inbred lines (RILs) under cold stress. A previously constructed high-density genetic linkage map was used for the mapping of the QTLs conferring cold tolerance at the plumule and seedling stages. A total of 4 QTLs for plumule-stage cold tolerance and 9 QTLs for seedling-stage cold tolerance were detected, with the maximum limit of detection reaching 5.20. Notably, a genetically overlapping QTL for both plumule and seedling stages was identified on chromosome 8, spanning a physical interval of 24 432 953-25 295 129 bp. Candidate genes within the detected QTL intervals were screened, and quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to analyze the gene expression during the plumule and seedling stages. The results revealed that LOC_Os03g06570, LOC_Os03g07100, LOC_Os06g08280, LOC_Os08g38440, LOC_Os08g39100, and LOC_Os08g39540 exhibited significantly differential expression between the parental lines. These genes were either significantly downregulated or upregulated under cold stress. Among them, the first three gene (LOC_Os03g06570, LOC_Os03g07100, and LOC_Os06g08280) were hypothesized to be key candidates regulating the cold tolerance of rice seedlings, while the latter three genes (LOC_Os08g38440, LOC_Os08g39100, and LOC_Os08g39540) were identified as comprehensive regulators of cold tolerance during both plumule and seedling stages. These findings lay a foundation for the fine mapping and cloning of cold tolerance genes at the plumule and seedling stages, providing valuable insights for breeding cold-tolerant rice varieties.
Quantitative Trait Loci/genetics* ; Oryza/growth & development* ; Seedlings/growth & development* ; Cold Temperature ; Chromosome Mapping ; Gene Expression Regulation, Plant