BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

Among the current curricula of medical teaching in China, most courses focus on the integrated teaching of a single organ-system combination, while there are relatively a few integrated courses that focus on the multiple dimensions between different organs and systems and between medical sciences and social sciences. In 2016, Chongqing Medical University started to cooperate with University of Leicester to establish the clinical medicine major and introduced the course of Integration for Clinical Application (ICA) that had been run well in University of Leicester for years. With reference to the education goal of our university, the curriculum group adopted a series of actions for the localization of this course from the aspects of teaching objectives, contents, teaching model, education resources, and quality of faculty. After the completion of the first round of this course, the passing rate reached 86.96%(100/115) in the quantified evaluation of accomplishment, which was higher than the passing rate of other courses introduced from University of Leicester. The quantitative expert assessment of this course also ranked among the top courses in our university, and student assessment showed that the ability indicators were improved by 25.00%- 38.00%. The above data show that good results have been achieved for the curriculum localization of ICA.

Background::The potential impact of pre-existing coronary artery stenosis (CAS) on acute pulmonary embolism (PE) episodes remains underexplored. This study aimed to investigate the association between pre-existing CAS and the elevation of high-sensitivity cardiac troponin I (hs-cTnI) levels in patients with PE.Methods::In this multicenter, prospective case-control study, 88 cases and 163 controls matched for age, sex, and study center were enrolled. Cases were patients with PE with elevated hs-cTnI. Controls were patients with PE with normal hs-cTnI. Coronary artery assessment utilized coronary computed tomographic angiography or invasive coronary angiography. CAS was defined as ≥50% stenosis of the lumen diameter in any coronary vessel >2.0 mm in diameter. Conditional logistic regression was used to evaluate the association between CAS and hs-cTnI elevation.Results::The percentage of CAS was higher in the case group compared to the control group (44.3% [39/88] vs. 30.1% [49/163]; P = 0.024). In multivariable conditional logistic regression model 1, CAS (adjusted odds ratio [OR], 2.680; 95% confidence interval [CI], 1.243–5.779), heart rate >75 beats/min (OR, 2.306; 95% CI, 1.056–5.036) and N-terminal pro-B type natriuretic peptide (NT-proBNP) >420 pg/mL (OR, 12.169; 95% CI, 4.792–30.900) were independently associated with elevated hs-cTnI. In model 2, right CAS (OR, 3.615; 95% CI, 1.467–8.909) and NT-proBNP >420 pg/mL (OR, 13.890; 95% CI, 5.288–36.484) were independently associated with elevated hs-cTnI. Conclusions::CAS was independently associated with myocardial injury in patients with PE. Vigilance towards CAS is warranted in patients with PE with elevated cardiac troponin levels.

Aim To investigate the protective effect of eriodictyol(ERD)on kidney damage in hypertensive mice and its possible mechanism.Methods Thirty C57BL/6J mice were randomly divided into five groups of six in each group.They were the control group(Ctrl),the hypertension model group(DOCA/Salt),the eriodictyol low-dose(DOCA/Salt+ERD-L),the high-dose(DOCA/Salt+ERD-H)and the positive drug perindopril group(DOCA/Salt+Perindopril).The classic hypertension model was induced by DOCA/Salt,and ERD was administered continuously for four weeks,and blood pressure,serum urea nitrogen,cre-atinine and other indicators were measured.Hematoxy-lin-eosin staining(HE)and Masson staining were used to observe the pathological changes of the kidney.Western blot was used to detect the expression of EMT marker proteins α-SMA,Vimentin,E-cadherin and FN in renal epithelial mesenchymal transition.Real-Time PCR was used to detect the expression of renal inflammatory cytokines Nlrp3,TNF-α,IL-1 β,IL-18,MCP-1 and NADPH oxidase(NOXs).Western blot was employed to detect the expression levels of TGF-β1,phosphorylated(p)-Smad2,TLR4 and NF-κB p65 proteins in mice.Results ERD intervention could delay the occurrence of hypertension,improve the pathological damage of renal tissue,reduce renal collagen deposition.It could also reduce the level of renal inflammation and oxidative stress,improve the level of EMT protein,and significantly reduce the pro-tein expression of TGF-β1,p-Smad2,TLR4 and NF-κB p65.Conclusions ERD is shown to have a signif-icant protective effect on DOCA/Salt hypertensive renal damage,and its mechanism may be related to the regu-lation of TGF-β1/Smad2,TLR4/NF-κB p65 and other signaling pathways.

Aim To investigate the effects of Cartham-us tinctorius L.extract(CTLE)on oxidative stress,lipid metabolism,and apoptosis levels of mice with al-cohol-induced liver injury and its mechanism of action.Methods The mouse model of alcohol-associated liver disease was established by chronic alcohol feeding and acute alcohol gavage.Mice were randomly divided into four groups.During the modeling period,the state changes of mice were observed every day,and their weight was recorded.At the end of modeling,blood and liver tissues were collected from each group of mice.The blood of mice was analyzed biochemically,and HE staining and Oil Red O staining were used to evaluate further the degree of pathological damage in the liver of mice.Quantitative real-time PCR(qPCR)and Western blot were applied to detect the mRNA and protein expression levels of p-PI3K,PI3K,p-Akt,Akt,p-mTOR,mTOR,p-FoxO1,FoxO1,p-FoxO3a,FoxO3a,p-FoxO4,FoxO4,BCL and BAX factors.Results Compared to the model group,the CTLE administration group showed improved hepatic patho-logical injury and reduced lipid deposition.The bio-chemical indexes in serum and liver,such as ALT,AST,TG,TC,and MDA levels were reduced,while GSH and SOD levels increased.Regulating the PI3K/Akt/FoxO pathway resulted in increased production of SOD,which reduced damage and apoptosis caused by reactive oxygen species(ROS).Conclusions CTLE can exert anti-oxidative stress and anti-apoptotic effects through the PI3K/Akt/FoxO pathway and attenuates alcoholic liver injury in mice,providing new ideas for the treatment of alcoholic liver disease and the develop-ment of related drugs.

Aim To explore the protective effects of Xiyanping injection against lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice,and investi-gate the underlying mechanism.Methods In the LPS-induced ALI mouse model,the protective effect of Xiyanping injection against ALI was evaluated by ob-serving the pathological indicators of lung tissue.Net-work pharmacology and molecular docking were used to explore its mechanism.Western blot method was used to validate the predicted target proteins.Results Xiy-anping injection significantly improved the pathological injury and alleviated inflammatory reactions in lungs of ALI mice.Four active ingredients were identified in Xiyanping injection,namely,14-deoxy-11-oxo-an-drographolide,14-deoxyandrographolide,14-deoxy-12-methoxyandrographolide,and andrographolide-19-β-D-glucoside.A total of 288 corresponding drug targets and 4 960 ALI-related targets were obtained,with 192 genes overlapping.The ten core targets associated with Xiyanping injection were identified as STAT3,EGFR,PIK3R1,MAPK1,PIK3CA,NFKB1,ESR1,MAPK8,JAK2,and FYN.GO enrichment analysis re-vealed 310 biological processes(BP),65 cellular components(CC),and 80 molecular functions(MF)associated with the overlapping genes.KEGG pathway enrichment analysis identified 141 pathways related to ALI,with the top 20 pathways including MAPK,TNF-α,VEGF,cAMP,mTOR,AMPK,NOD,JAK-STAT,IL-17,and NF-κB.Molecular docking results demonstrated strong binding affinity between core tar-gets(MAPK1,MAPK8,NFKB1)and active ingredi-ents(14-deoxy-12-methoxyandrographolide and 14-de-oxyandrographolide).Western blotting showed that medium and high doses of Xiyanping injection signifi-cantly downregulated p38,JNK,ERKl/2,NF-κB p65 protein expression in lung tissue of ALI mice(P＜0.01).Conclusions Xiyanping injection has a cer-tain protective effect against ALI,and the mechanism is related to regulating MAPK and NF-κB signaling pathways.

Objective:To summarize the best evidence for home self-volume monitoring in patients with chronic heart failure (CHF), so as to provide evidence for accurate monitoring of volume status.Methods:Computer search was conducted for literature on home self-volume monitoring in CHF patients in UpToDate, BMJ best practice, Guidelines International Network, Cochrane Library, PubMed, Embase, CNKI, Wanfang Data, China Biology Medicine disc and other websites and databases. The search period was from establishment of databases to August 31, 2023. After the quality evaluation of various literatures, the evidence was extracted and summarized.Results:A total of 21 literatures were included, and 24 pieces of evidence were summarized from 4 aspects, including monitoring objectives, monitoring content and methods, health education and resource integration and support.Conclusions:This study summarizes the best evidence for home self-monitoring of CHF patients, which is beneficial for nursing staff to provide a basis for providing patients with home self-monitoring plans, thereby improving the accuracy and effectiveness of patient self-monitoring.

Objective To explore the efficacy and safety of low-dose tirofiban in stent-assisted coil embolization(SAC)for ruptured intracranial aneurysms.Methods From April 2011 to September 2020,335 patients of ruptured intracranial aneurysms with subarachnoid hemorrhage(SAH)admitted in the First People's Hospital of Changzhou were retrospectively analyzed.All cases underwent stent-assisted coil embolization within 24-48 h and antiplatelet medications.The patients were divided into dual antibody group(89 cases)and tirofiban group(246 cases).Baseline and clinical data of all patients were collected for comparison between groups,including age,sex,hypertension,diabetes mellitus,Hunt-Hess grade at admission,modified Fisher scale score at admission,aneurysm diameter(＞5 mm,≤5 mm),aneurysm location(anterior circulation,posterior circulation),postoperative acute hydrocephalus or intraventricular hemorrhage,postoperative complete embolization rate of ruptured aneurysm.All patients with ruptured intracranial aneurysm with SAH were confirmed by emergency cerebral CT scan after admission.The Raymond grading criteria were used to evaluate the embolization effect after operation:grade Ⅰ refers to no development(complete embolization),grade Ⅱ refers to only aneurysm neck development(incomplete embolization),and grade Ⅲ refers to aneurysm body development,in which Raymond grading Ⅰ orⅡ indicates effective embolization.Tirofiban group:4.2 μg/kg tirofiban was intravenously injected after the coil was placed in the aneurysm lumen and the stent was released,followed by maintenance dose 0.07 μg/(kg·min)for 6-8 h,and aspirin 100 mg and clopidogrel 75 mg were given as sequential dual antiplatelet therapy 2 hours before the tirofiban infusion was stopped.Dual antiplatelet group:a loading dose of aspirin 300 mg and clopidogrel 300 mg was given at least 2 hours before stent implantation,and then transferred to aspirin 100 mg and clopidogrel 75 mg given on the second day after operation.All patients received aspirin(100mg/d)for 6 months and clopidogrel(75 mg/d)for 3 months after operation.The efficacy indicators,safety indicators,adverse events and other complications of the two groups were collected and compared.The efficacy indicators were the incidence of thrombotic events during operation and within 72 hours after operation.The safety indicators were the incidence of intraoperative and early postoperative intracranial hemorrhage(within 48 hours after operation),the incidence of late postoperative intracranial hemorrhage(over 48 hours after operation),and the incidence of intracranial hemorrhage related to external ventricular drainage(symptomatic and asymptomatic).The adverse event was the occurrence of drug-related thrombocytopenia.Other complications were delayed ischemic events.The modified Rankin scale(mRS)score was used to evaluate the clinical prognosis of patients at 180 days after operation.mRS score ≤2 was defined as good prognosis,mRS score＞2 was defined as poor prognosis,of which 6 was defined as death.Results(1)There were no significant differences in baseline and clinical data between the tirofiban group and the dual antibody group(all P＞0.05).(2)There was no significant difference in the proportion of patients with good outcome(75.2％[185/246]vs.74.2％[66/89],P=0.845)and death(10.2％[25/246]vs.12.4％[11/89],P=0.566)at 180 days after operation between the tirofiban group and the dual antiplatelet group.(3)There was no significant difference in the incidence of intraoperative(0.8％[2/246]vs.4.5％[4/89],P=0.075)and postoperative thrombotic events(11.0％[27/246]vs.13.5％[12/89],P=0.527)between the tirofiban group and the dual antiplatelet group.(4)Results about safety comparison between this two antiplatelet regimens showed that the incidence of early postoperative intracranial hemorrhage were lower in the tirofiban group than that in the dual antiplatelet group(2.8％[7/246]vs.10.1％[9/89],P=0.014).There were no significant differences in the symptomatic external ventricular drainage related intracranial hemorrhage(0 vs.2/15,P=0.050),incidences of intraoperative intracranial hemorrhage(1.6％vs.3.4％,P=0.580),late postoperative intracranial hemorrhage(3.3％vs.4.5％,P=0.836),and drug-related thrombocytopenia(0.4％vs.1.1％,P=0.461)between the two groups.Conclusion Low-dose tirofiban infusion in SAC for ruptured aneurysms may prevent perioperative thromboembolic events without high risk of intracranial hemorrhage.

Objective:To investigate the expression and clinical significance of CD30 in patients with diffuse large B-cell lymphoma(DLBCL).Methods:A retrospective analysis was conducted on 124 cases of primary DLBCL diagnosed at Changzhou Second People's Hospital Affiliated with Nanjing Medical University from January 2018 to July 2020.The expression of CD30 in patients with DLBCL was detected by immunohistochemical method,and the clinicopathological characteristics were analyzed and compared between CD30+and CD30-groups.Kaplan-Meier analysis was used for survival analysis.The relationship between CD30 expression and clinical features and prognosis were analyzed.Results:Among the 124 patients with DLBCL,19 patients expressed CD30,and the positive rate is 15.32％.The clinico-pathological characteristics of CD30+in patients with DLBCL were characterized by low age,more common in males,fewer extranodal lesions,lower international prognostic index(IPI),GCB type being more common in Hans subtype,and achieving better therapeutic effects(P＜0.05).However,there were no significant statistical differences in B-symptoms(P=0.323),Ann Arbor staging(P=0.197),Eastern Cooperative Oncology Group(ECOG)score(P=0.479),lactate dehydrogenase(LDH)(P=0.477),and the involvement of bone marrow(P=0.222).There were significant differences in OS and PFS between the CD30+and CD30-groups(x2=5.653,P=0.017;x2=4.109,P=0.043),the CD30+group had a better prognosis than that of the CD30-group.The results of subgroup analysis showed that the CD30+group in the IPI score=1-2,LDH elevated group had a better prognosis(P＜0.05).In the subgroups of Ann Arbor staging Ⅲ-Ⅳ(P=0.055)and non GCB type(P=0.053),the CD30+group had a good prognosis trend,but the difference was not statistically significant.The results of univariate analysis showed that the good prognosis of DLBCL patients was closely related to CD30+expression,no B-symptoms,early Ann Arbor staging,low ECOG score,normal LDH,low IPI score,fewer extranodal involvement,and obtaining the best therapeutic effect as CR(all P＜0.05).COX multivariate regression analysis showed that the presence of B-symptoms and achieving the best therapeutic effect as Non-CR were independent risk factors affecting the prognosis of DLBCL patients(P＜0.05).Conclusion:The CD30+expression in DLBCL patients indicates a good prognosis and has certain diagnostic value in evaluating the prognosis of DLBCL patients.

Objective:RS4;11 cell line was used to establish BCL-2 inhibitor-resistant cell lines of B-cell acute lymphoblastic leukemia(B-ALL)and explore the possible mechanisms of drug resistance.Methods:RS4;11 cell line was continuously induced and cultured by low and ascending concentrations of BCL-2 inhibitors navitoclax and venetoclax to construct navitoclax-resistant cell line RS4;11/Nav and venetoclax-resistant cell line RS4;11/Ven.The cell viability was detected by MTT assay,and the cell apoptosis was detected by flow cytometry.Differentially expressed genes(DEGs)between RS4;11 drug-resistant cell lines and parental cell line were detected by transcriptome sequencing technology(RNA-seq),and mRNA expression levels of DEGs between drug-resistant cell lines and parental cell line were detected by real-time PCR(RT-PCR).Western blot was used to detect the expression levels of BCL-2 family anti-apoptotic proteins in drug-resistant cell lines and parental cell line.Results:The drug-resistant cell lines RS4;11/Nav and RS4;11/Ven were successfully established.The resistance index(RI)of RS4;11/Nav to navitoclax and RS4;11/Ven to venetoclax was 328.655±47.377 and 2 894.027±300.311,respectively.The results of cell apoptosis detection showed that compared with the drug-resistant cell lines,RS4;11 parental cell line were significantly inhibited by BCL-2 inhibitors,while the apoptosis rate of drug-resistant cell lines was not affected by the drugs.Western blot assay showed that the expression of anti-apoptotic proteins of BCL-2 family did not increase significantly in drug-resistant cell lines.RNA-seq,RT-PCR and Western blot assays showed that the expression of EP300 in drug-resistant cell lines was significantly higher than that in parental cell line(P＜0.05).Conclusion:Drug-resistant B-ALL cell lines could be successfully established by exposing RS4;11 cell line to the ascending concentration of BCL-2 inhibitors,and the drug resistance mechanism may be related to the overexpression of EP300.