BACKGROUND:LncRNA-TNFRSF13C,an important factor in B cell development and function,is expressed in periodontal tissues of patients with periodontitis,but the specific mechanism is still unclear. OBJECTIVE:To investigate the mechanism of lncRNA-TNFRSF13C regulating miR-1246 on hypoxia-inducible factor 1α in periodontal cells. METHODS:Human periodontal ligament cells(hPDLCs)were treated with lipopolysaccharide and divided into group A(hPDLCs cell lines without transfection),group B(hPDLCs cell lines transfected with TNFRSF13C NC-siRNA),group C(hPDLCs cell lines transfected with TNFRSF13C-siRNA),group D(hPDLCs cell line transfected with miR-1246 mimics),group E(hPDLCs cell line transfected with miR-1246 siRNA),group F(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 mimics),and group G(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 siRNA).The relative expression of lncRNA-TNFRSF13C and miR-1246 in each group was detected by qRT-PCR.Cell counting kit-8 assay was used to detect cell viability.Apoptosis was detected by flow cytometry.Expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins was detected by western blot.The correlation between lncRNA-TNFRSF13C and miR-1246 was analyzed by Pearson,and the targeting relationship was analyzed by dual-luciferase reporter assay. RESULTS AND CONCLUSION:There was no significant difference in human periodontal ligament cell activity,apoptosis rate and protein indexes between groups A and B(P>0.05).Compared with group B,hPDLCS cell activity in group C was increased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were decreased(P<0.05).Compared with group C,hPDLCS cell activity in group D was decreased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were increased(P<0.05).Compared with group D,the cell activity of group E was increased(P<0.05).The cell activity in group F was lower than that in group E,and the apoptosis rate was reduced in both groups E and F(P<0.05).Compared with group F,the cell activity of group G was increased,and the apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor were decreased(P<0.05).LncRNA-TNFRSF13C was positively correlated with miR-1246(P<0.05).Compared with the TNFRSF13C-siRNA group,the fluorescence activity of miR-1246-wt in the TNFRSF13C-NC group was reduced(P>0.05);compared with the miR-1246-NC group,the fluorescence activities of hypoxia-inducible factor 1α-wt and vascular endothelial growth factor-wt in the miR-1246 mimics group were increased(P<0.05).To conclude,down-regulation of lncRNA-TNFRSF13C can promote the activity of periodontal cells treated with lipopolysaccharide,reduce apoptosis,and inhibit hypoxia-inducible factor 1α and vascular endothelial growth factor.The mechanism is related to the regulation of miR-1246 activity.

OBJECTIVE To explore comprehensive management and potential issues associated with long-term prescriptions medications of psychiatry， in order to provide a reference for the comprehensive management of long-term prescriptions of psychiatry in psychiatric hospitals and other medical institutions’ pharmacies. METHODS Starting from the applicable principles for long-term prescriptions of psychiatry， this study introduced the standardized assessment and precautions before issuing long-term prescriptions， the formulation and adjustment of the drug list， as well as the rational management of the long-term prescriptions. It also analyzed potential issues that may arise in the comprehensive management of long-term prescription medications and proposed corresponding countermeasures and suggestions. RESULTS & CONCLUSIONS Prior to initiating long-term prescriptions， a standardized assessment should be conducted on patients from the aspects of their psychiatric condition and long-term potential risk factors， pharmacological treatment plans and other non-pharmacological therapies， physical illnesses. Additionally， healthcare providers should fulfill their obligation to inform patients or their family members. The comprehensive management of long-term prescription medications should be jointly established and improved by multiple departments， and the formulation of drug catalogs should avoid including drugs with potential social harm or medication risks while complying with policy requirements. Furthermore， measures such as adding special identifiers to long-term prescriptions， providing patients with reminders about （No.YGLX202537） prescription expiration， or offering online consultations can also effectively enhance the rationality of medication use under long-term prescriptions. Currently， the implementation of long-term prescriptions in psychiatry remains challenged by inconsistencies in prescription duration， incomplete coverage of diagnostic categories， poor patient adherence， and the risk of deviation in clinical assessments. In this regard， measures such as collaborating with multiple departments to strengthen long-term prescription information management， providing matching pharmaceutical services， ensuring the quality and rationality of long-term prescription implementation， and using modern methods to screen high-risk patients can be taken to improve patient medication compliance and safety.

ObjectiveTo investigate the impacts of three types of acupuncture timing factors， including intervention initiation time， needle retention duration， and treatment intervals， on the treatment efficacy of ischemic stroke （IS） and provide evidence-based support for optimizing acupuncture timing strategies. MethodsA systematic search was conducted in both Chinese and English databases， including CNKI， Wanfang Data， VIP， CBM， PubMed， Web of Science， Cochrane Library， and Embase. The randomized controlled trials involving multiple acupuncture intervention time groups were included to reduce inter-study heterogeneity. A network meta-analysis model was constructed to compare the effects of different intervention initiation time， needle retention duration， and treatment intervals on primary IS outcomes， including total clinical response rate， neurological and limb function scores， quality of life score， and hemorheological parameters. Adverse events were also assessed. ResultsA total of 32 studies were included. The results showed that early acupuncture intervention （1-3 days and 4-7 days after onset） outperformed late intervention （>14 days） in terms of response rate， neurological function improvement， and quality of life （P<0.05， P<0.01）. Extending needle retention time to 40-60 min was associated with improved response rates and hemorheological parameters （P<0.01）. No significant differences were observed among different treatment intervals. The evidence of key outcomes showed high overall certainty， with low heterogeneity and inconsistency. Sensitivity analyses and publication bias assessments indicated the stability of the main results and a low risk of publication bias. No significant adverse reaction was reported. ConclusionDifferent timing factors significantly influence the efficacy of acupuncture treatment for IS. Early intervention and appropriately extended needle retention time result in better clinical outcomes， while treatment interval has no significant effect on efficacy. The inclusion of studies with multiple acupuncture timing groups helps control methodological heterogeneity and enhances the robustness of the analysis， providing evidence-based guidance for optimizing acupuncture timing strategies in IS management.

Background::The ability to generate functional hepatocytes without relying on donor liver organs holds significant therapeutic promise in the fields of regenerative medicine and potential liver disease treatments. Clustered regularly interspaced short palindromic repeats (CRISPR) activator (CRISPRa) is a powerful tool that can conveniently and efficiently activate the expression of multiple endogenous genes simultaneously, providing a new strategy for cell fate determination. The main purpose of this study is to explore the feasibility of applying CRISPRa for hepatocyte reprogramming and its application in the treatment of mouse liver fibrosis.Method::The differentiation of mouse embryonic fibroblasts (MEFs) into functional induced hepatocyte-like cells (iHeps) was achieved by utilizing the CRISPRa synergistic activation mediator (SAM) system, which drove the combined expression of three endogenous transcription factors— Gata4, Foxa3, and Hnf1a—or alternatively, the expression of two transcription factors, Gata4 and Foxa3. In vivo, we injected adeno-associated virus serotype 6 (AAV6) carrying the CRISPRa SAM system into liver fibrotic Col1a1-Cre ER; Cas9 fl/fl mice, effectively activating the expression of endogenous Gata4 and Foxa3 in fibroblasts. The endogenous transcriptional activation of genes was confirmed using real-time quantitative polymerase chain reaction (RT-qPCR) and RNA-seq, and the morphology and characteristics of the induced hepatocytes were observed through microscopy. The level of hepatocyte reprogramming in vivo is detected by immunofluorescence staining, while the improvement of liver fibrosis is evaluated through Sirius red staining, alpha-smooth muscle actin (α-SMA) immunofluorescence staining, and blood alanine aminotransferase (ALT) examination. Results::Activation of only two factors, Gata4 and Foxa3, via CRISPRa was sufficient to successfully induce the transformation of MEFs into iHeps. These iHeps could be expanded in vitro and displayed functional characteristics similar to those of mature hepatocytes, such as drug metabolism and glycogen storage. Additionally, AAV6-based delivery of the CRISPRa SAM system effectively induced the hepatic reprogramming from fibroblasts in mice with live fibrosis. After 8 weeks of induction, the reprogrammed hepatocytes comprised 0.87% of the total hepatocyte population in the mice, significantly reducing liver fibrosis. Conclusion::CRISPRa-induced hepatocyte reprogramming may be a promising strategy for generating functional hepatocytes and treating liver fibrosis caused by hepatic diseases.

Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.

In this study,the immunological characteristics of the PhoP protein were explored with a two-component system of Mycobacterium tuberculosis(Mtb).Bioinformatics was used to predict the B and T cell epitopes of the PhoP protein.A re-combinant expression plasmid was constructed by PCR analysis of the phoP sequence and cloning into the prokaryotic expres-sion vector pET-28a(+).Competent Escherichia coli BL21 cells were transformed with the recombinant plasmid and expres-sion was induced with IPTG.The recombinant PhoP protein was purified by affinity chromatography.Serum levels of PhoP-specific antibodies in Mtb-infected mice and tuberculosis(TB)patients were analyzed with an ELISA.BALB/c mice were im-munized with the PhoP recombinant protein by intramuscular injection.Sera of mice were collected and antibody titers were detected with an ELISA and specificity was assessed by West-ern blot analysis.Mouse splenocytes were isolated and the pro-portions of IFN-y-positive cells and cytokine levels were detec-ted with an ELISpot and ELISA,respectively.Bioinformatics i-dentified 24 B cell and 11 T cell epitopes of the PhoP protein.A prokaryotic recombinant vector of PhoP was successfully con-structed and the recombinant PhoP protein was obtained by purification.Specific antibody levels to PhoP in sera of Mtb in-fected mice and TB patients increased significantly,with preci-sion of 99.9％and 82.5％,and specificity of 100％,respectively.PhoP protein immunization successfully induced production of specific antibodies in mice.Stimulated by antigens in vitro,IL-2 and IFN-γ levels were significantly increased in the splenocytes of immunized mice.Immunization with the PhoP protein induce a humoral immune response and Thl-dominated cellular immu-nity,indicating that the PhoP protein was immunogenic with diagnostic efficacy for TB.These results lay a foundation to clari-fy the role of PhoP in Mtb infection and application for diagnosis and prevention of TB.

In order to understand the potential target and related mechanism of action of Termina-lia Chebula treatment,network pharmacology and molecular docking methods were used in this experiment,and the challenge test of Salmonella from yak was performed.The active ingredients and potential targets of Terminalia Chebula were screened through HERB cluster identification database,TCMSP database and SwissTargetPrediction web page tool,and"gastroenteritis"was searched through OMIM and GeneCards database.Cytoscape and STRING databases were used to construct the Terminalia Chebula PPI network to screen out key targets,the intersection targets between Terminalia Chebula and enteritis were obtained through Venny platform,and gene ontol-ogy(GO)and Kyoto encyclopedia database of genes and genomics(KEGG)were enriched through DAVID database.The core target of screening was verified by molecular docking.After that,the gastrointestinal inflammation model of mice was established,the pathological changes of gastroin-testinal tract were observed,and the effect of Terminalia Chebula on the target protein was veri-fied by Western blot test.The results showed that:after analyzing and sorting out 8 main active in-gredients of Terminalia Chebula,118 targets of Terminalia Chebula were screened,11 161 targets of gastroenteritis and 100 targets of intersection were obtained;the core targets of PTGS2,CASP3,SLC3A2,Bax,Bcl-2 and TP53 of Terminalia Chebula and enteritis were obtained through PPI network.GO and KEGG enrichment analysis collected 337 items and 138 items,respectively,mainly related to chemokine pathway,PI3K-Akt signaling pathway,apoptosis related pathway,i-ron ion transport related pathway,NF-κB signaling pathway,etc.The results of molecular docking showed that chebulidic acid,the first active component of chebulidic acid,can bind to Bax,Bcl-2,PTGS2 and SLC3A2 through hydrogen bonding,hydrophobic action,π-π packing force and other intermolecular forces.The pathological tissue sections showed that Terminalia Chebula could sig-nificantly recover gastrointestinal tissue injury.Western blot test results showed that Terminalia Chebula can regulate the process of apoptosis and iron death through Bax/Bcl-2 and PTGS2/SLC3A2 pathways to achieve the effect of treating intestinal inflammatory damage.The results showed that Terminalia Chebula can regulate the occurrence and development of enteritis by regu-lating apoptosis and iron death through Bax/Bcl-2 and PTGS2/SLC3A2 pathways.Terminalia Chebula has the characteristics of multi-target and multi-pathway in the treatment of enteritis.

Objective:To analyze the expression of MCP-1 and CCR2 in newly diagnosed diffuse large B-cell lymphoma(DLBCL),and to evaluate their correlation with clinicopathological features and prognosis.Methods:A total of 141 patients with DLBCL diagnosed and treated in the Department of Hematology,the First Affiliated Hospital of Bengbu Medical College from January 2017 to May 2022 were retrospectively collected.The clinical characteristics,pathological data and prognostic factors of the patients were collected.Immunohistochemical staining was used to detect the expression of MCP-1 and CCR2 in the tissues of newly treated DLBCL patients,and to analyze the relationship between MCP-1 and clinical characteristics,prognosis and survival of patients.Results:The expression of MCP-1 and CCR2 were correlated with Ann Arbor stage,IPI score,lactate dehydrogenase(LDH),Ki-67 index and therapeutic effect.There were no significant correlation between the expression of MCP-1 or CCR2 and other clinical histopathological parameters such as gender,age,β2-microglobulin,BCL-2,BCL-6,Hans classification,initial location,B symptoms,bone marrow involvement.There was a statistical difference in OS and PFS between the MCP-1 or CCR2 positive group and the negative group,which was associated to poor prognosis.Univariate Cox regression analysis showed that β2-microglobulin,Ki-67 index,IPI score,MCP-1,CCR2 expression levels and disease remission affected the PFS and OS of DLBCL patients(P＜0.05).Gender,age,LDH,BCL-2,BCL-6,Hans classification,primary tumor site,B symptoms,bone marrow involvement,Ann Arbor stage had no effect on PFS and OS(P＞0.05).Multivariate analysis showed that β2-microglobulin,Ki-67 index,IPI score,MCP-1,CCR2 expression levels and disease remission were independent influencing factors of patients(P＜0.05).Conclusion:The expression rate of MCP-1 or CCR2 in newly treated DLBCL is high,and it is correlated with the clinical features of poor prognosis such as stage and LDH of DLBCL patients,which is a poor prognostic factor affecting PFS and OS.

Objective:To investigate the expression level of urothelial carcinoembryonic antigen 1(lncRNA UCA1)in the bone marrow of acute myeloid leukemia(AML)patients,and to explore the clinical significance of lncRNA UCA1 expression level in AML patients.Methods:Bone marrow samples of 50 AML patients were collected as experimental group,and bone marrow samples of 20 iron deficiency anemia(IDA)patients were collected as control group.The relevant clinicopathological characteristics of AML patients were collected.Real-time quantitative PCR(qRT-PCR)was used to detect the expression level of lncRNA UCA1 in the experimental and control groups,and the relationships between lncRNA UCA1 expression and clinical pathological characteristics and prognosis in AML patients were analyzed.Kaplan-Meier curves were used to analyze the effect of lncRNA UCA1 on the overall survival(OS)of AML patients;And Cox regression model was used to analyze the factors affecting the prognosis of AML patients.Results:Compared with the control group,the expression level of lncRNA UCA1 was significantly elevated in patients with AML(P＜0.001);The proportion of patients with hemoglobin lower than 90 g/L in lncRNA UCA1 high expression group was significantly higher than that in lncRNA UCA1 low expression group(P=0.004);The expression level of lncRNA UCA1 was higher in M1,M2,and M4 subtypes,while it was lower in M0 and M5 subtypes,and the difference was statistically significant(P=0.009).There were no significant difference in sex,age,white blood cell(WBC)count,platelet(PLT)count,bone marrow blasts,chemotherapy regimen and efficacy,karyotype,gene mutation,and prognostic risk stratification between patients in UCA1 high expression group and those in UCA1 low expression group(all P＞0.05).The OS of patients with high expression of lncRNA UCA1 was significantly shorter than that of patients with low expression of lncRNA UCA1(P=0.0229).Conclusion:The expression level of lncRNA UCA1 is significantly upregulated in AML patients.High expression of lncRNA UCA1 is associated with poor clinicopathological features and poor prognosis.Therefore,lncRNA UCA1 can be used as a prognostic indicator and a potential therapeutic target for AML patients.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT)is one of the main treatment methods for hematological malignancies.With the continuous improvement and popularization of transplantation technology,it has brought hope for prolonging the lives and improving the survival rate of patients with hematological malignancies. However,postoperative invasive fungal infection (IFI)is the most common infectious complication and the main cause of death,with difficult early diagnosis and extremely high mortality.This paper summarizes the latest research progress on the pathogenic types,diagnostic methods,high-risk factors and treatment regimen of Candida,Aspergillus and Mucor associated with postoperative IFI,which is expected to provide references for improving the early diagnosis rate and treatment effectiveness of postoperative IFI.