ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

[摘 要] 目的：探讨同源异型盒蛋白D11（HOXD11）对Ki-67的转录调控作用及其对喉鳞状细胞癌（LSCC）细胞恶性生物学行为的影响和机制。方法：结合高通量测序数据，通过GEO、UALCAN数据库分析HOXD11在LSCC中差异表达。收集2022年1月至2025年1月联勤保障部队第九八〇医院手术切除的60例LSCC患者的癌及癌旁组织标本，以及人LSCC细胞系AMC-HN-8、TU-177、TU-686和人正常喉上皮细胞（HNLEC），构建敲低或过表达HOXD11的细胞系，将细胞分为对照组、HOXD11敲低组及HOXD11过表达组。通过RT-qPCR法检测HOXD11和Ki-67基因在LSCC组织和细胞中mRNA表达水平，免疫组织化学（IHC）法分析两者在LSCC组织中的蛋白表达及分布，WB法进一步验证蛋白差异表达。采用MTS、克隆形成实验及Transwell实验分别检测敲低或过表达HOXD11对LSCC细胞增殖、迁移与侵袭的影响。通过双萤光素酶报告基因实验和染色质免疫沉淀（ChIP）实验验证HOXD11对Ki-67启动子活性的调控作用。结果：GEO和UALCAN数据库分析表明，HOXD11在LSCC中高表达（P < 0.01）。在LSCC组织中HOXD11和Ki-67 mRNA和蛋白表达均显著高于癌旁组织（均P < 0.01），同时，两者表达水平之间存在正相关（r = 0.26，P < 0.05）；LSCC细胞系中HOXD11 mRNA表达显著高于HNLEC（均P < 0.01）。敲低HOXD11显著抑制LSCC细胞的增殖、迁移和侵袭能力（均P < 0.01），而过表达HOXD11则促进细胞的这些恶性生物学行为（均P < 0.01）。双萤光素酶报告基因实验及ChIP实验均证实，HOXD11可直接结合到Ki-67启动子区上，调控其表达（P < 0.01）。回复实验显示，过表达Ki-67可部分逆转敲低HOXD11对LSCC细胞增殖、迁移与侵袭的抑制作用（均P < 0.01）。结论：HOXD11在LSCC组织及细胞系中均呈高表达，其机制在于通过直接调控Ki-67的转录活性，从而增强LSCC细胞的增殖、迁移及侵袭能力。

Objective To explore the action mechanism of Bruton's tyrosine kinase (BTK) inhibitor zanubrutinib on renal fibrosis after renal ischemia-reperfusion injury (RIRI). Methods C57BL/6 mice were randomly divided into sham operation group, modeling group and modeling + zanubrutinib treatment group (zanubrutinib group), with 5 mice in each group. The groups underwent sham operation, RIRI modeling and RIRI modeling + zanubrutinib (5 mg/kg) treatment, respectively. Tissues were collected after 21 days. The morphological changes of the kidneys, histopathological changes, levels of M1 macrophages in the kidneys, inflammatory responses in the kidneys, and the expression of related inflammatory signaling pathways of macrophages induced by lipopolysaccharide(LPS) + interferon(IFN)-γ in vitro after lentivirus transfection were observed. Results Compared with the sham operation group, the kidneys of the modeling group mice shrank, the ratio of unilateral kidney weight to mouse body weight decreased, renal tubular interstitial fibrosis worsened, and the expression of α-smooth muscle actin (α-SMA) and type I collagen in the kidneys increased. The expression of F4/80 and CD86 in the kidneys increased, the lumen of the renal proximal convoluted tubules was significantly dilated, cellular debris accumulated in the lumen and inflammatory cell infiltration occurred, and the messenger RNA (mRNA) levels of CD86, tumor necrosis factor (TNF)-α, interleukin (IL)-6, inducible nitric oxide synthase (iNOS) and IL-1β in the kidneys increased. Compared with the modeling group, the kidneys of the zanubrutinib group mice enlarged after RIRI, the ratio of unilateral kidney weight to mouse body weight increased, renal tubular interstitial fibrosis was alleviated, and the expression of α-SMA and type I collagen in the kidneys decreased. The expression of F4/80 and CD86 in the kidneys decreased, the number of CD45⁺ lymphocytes and CD11b⁺ F4/80⁺ macrophages in the kidneys decreased, the infiltration of CD11b⁺ F4/80⁺ and CD86⁺ macrophages in the damaged tissue decreased, the degree of renal inflammatory pathological changes was milder, and the mRNA levels of CD86, TNF-α, IL-6, iNOS and IL-1β in the kidneys decreased. In vitro experiments using LPS+IFN-γ to induce M1-type macrophages found that the phosphorylation levels of phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), and nuclear factor (NF)-κB increased, while the phosphorylation levels decreased after BTK knockdown, indicating that BTK knockdown may inhibit the PI3K/Akt and NF-κB related inflammatory signaling pathways, thereby reducing the pro-inflammatory effects of LPS+IFN-γ induced M1-type macrophages. Conclusions Zanubrutinib may alleviate renal fibrosis after RIRI by inhibiting the PI3K/Akt and NF-κB related inflammatory signaling pathways, reducing the infiltration of M1 macrophages and the expression of related inflammatory factors, providing potential evidence for its clinical application.

OBJECTIVE: To explore the predictive efficacy of the early lactic acid/albumin ratio (LAR) for the occurrence of acute skin failure (ASF) in patients with sepsis. METHODS: A retrospective study was conducted to collect the clinical data of 115 patients with sepsis admitted to the intensive care unit (ICU) of the First Affiliated Hospital of Dalian Medical University from June 2022 to March 2024. The patients' gender, age, length of ICU stay, past medical history, and severity scores, use of mechanical ventilation or vasoactive drugs, albumin (Alb), lactic acid (Lac), mean arterial pressure (MAP), and blood gas analysis indicators within 24 hours of ICU admission were collected, and LAR was calculated. The patients were divided into two groups based on whether they developed ASF, and the clinical data between the two groups were compared. Multivariate Logistic regression analysis was used to screen the risk factors for the occurrence of ASF in patients with sepsis. The receiver operator characteristic curve (ROC curve) was drawn to analyze the predictive value of LAR for the occurrence of ASF in patients with sepsis. RESULTS: A total of 115 patients with sepsis were enrolled in the final analysis, among whom 35 developed ASF and 80 did not. The incidence of ASF was 30.43%. Univariate analysis showed that compared with the non-ASF group, the ASF group had higher acute physiology and chronic health evaluation II (APACHE II) score, proportion of using vasoactive drugs, Lac, and LAR as well as lower Alb and MAP, with statistically significant differences. Multivariate Logistic regression analysis was conducted on the factors with statistical significance in the univariate analysis, and the results showed that Alb [odds ratio (OR) = 0.639, 95% confidence interval (95%CI) was 0.474-0.862, P = 0.003], Lac (OR = 17.228, 95%CI was 1.517-195.641, P = 0.022), MAP (OR = 0.905, 95%CI was 0.855-0.959, P = 0.001), and LAR (OR < 0.001, 95%CI was < 0.001-0.005, P = 0.033) were independent risk factors for the occurrence of ASF in patients with sepsis. ROC curve analysis showed that the area under the ROC curve (AUC) of LAR for predicting the occurrence of ASF in patients with sepsis was 0.867 (95%CI was 0.792-0.943), which was superior to Alb, Lac, and MAP [AUC (95%CI) was 0.739 (0.648-0.829), 0.844 (0.760-0.929), and 0.860 (0.783-0.937), respectively]. When the optimal cut-off value of LAR was 0.11, the sensitivity was 65.7%, the specificity was 96.3%, and the Youden index was 0.620. Patients were grouped based on the optimal cut-off value of LAR, and the results showed that the incidence of ASF in the LAR > 0.11 group was significantly higher than that in the LAR ≤ 0.11 group [88.89% (24/27) vs. 12.50% (11/88), P < 0.05]. CONCLUSIONS LAR has early predictive value for the occurrence of ASF in patients with sepsis, and its efficacy is superior to that of Lac or Alb alone.
Humans ; Sepsis/blood* ; Retrospective Studies ; Lactic Acid/blood* ; Male ; Female ; Intensive Care Units ; Middle Aged ; Risk Factors ; Predictive Value of Tests ; Serum Albumin/analysis* ; ROC Curve ; Aged

In the context of aging population, the issue of polypharmacy among elderly patients with mental disorders has become increasingly prominent.Cognitive decline and depressive symptoms render these patients more vulnerable to medication-related risks, while poorly managed physical illnesses further complicate their treatment.To address these challenges, this paper proposes a series of management strategies that emphasize the critical role of pharmacists in conducting medication reviews.A comprehensive assessment of drug risks, benefits, and patient adherence is essential.The proposed strategies not only require careful consideration of patients' clinical needs and individual preferences but also highlight the importance of multidisciplinary team collaboration to reach a consensus on medication therapy.The use of clinical decision support systems as an auxiliary tool is recommended to enhance the scientific rigor of medication decision-making.Furthermore, pharmacists can optimize medication regimens through scientifically validated methods and promote patient or family involvement in self-management to improve acceptance and adherence to treatment adjustments.

The rabies virus(RABV)that causes rabies mainly attacks the peripheral and central nervous systems.In the later stages of infection,it is scattered in the salivary glands and transmit-ted to other susceptible animals through infectious saliva.To study dispersion of the RABV in the three pairs of salivary gland tissues,the street strain PB4 of the RABV was inoculated into 21-day-old female mice through the hind limb muscles.During the moribund stage of the mice,the sublin-gual gland,submandibular gland and parotid gland were collected,respectively.The TCID50 titer of RABV in the three kinds of glands of the mice and the copy number of the RABV N gene were de-tected,and RABV in different salivary glands was observed by immunofluorescence.The results showed that PB4 was dispersed in all three kinds of salivary glands of the mice,with the largest a-mounts in the parotid gland,followed by the submandibular gland,and the lowest amount in the sublingual gland.Three-month-old dogs were inoculated with PB4 through the cranial cavity,and saliva were collected every 12 h after inoculation.The saliva samples were detected by TCID50 and RT-qPCR.And during the moribund stage of the dogs when the disease occurred,the three pairs of salivary glands were collected.Through the determination of the TCID50 titer,RT-qPCR and immu-nofluorescence detection,it was demonstrated that among the three different salivary glands of the dogs,the largest amount of PB4 was found in the parotid gland and the lowest in the sublingual gland.Our results in mice and dogs clearly proved that the parotid gland was consistently found to exhibit the highest content of street RABV among the three major salivary glands,which could en-rich experimental data for analyzing the dispersion of RABV in the salivary glands and interpreta-tion of the intermittent secretion of saliva in clinically rabid dogs.

Objective By exploring the establishment of a follow-up and pre-visit model,this study aims to facilitate the development of a modernized Chinese medical service model for public hospitals,characterized by optimized workflows,continu-ous care models,efficient services,comfortable environments,and patient-centered attitudes,thereby improving patient experi-ence and enhancing the quality of outpatient follow-up visits.Methods Under the value co-creation framework,with both healthcare providers and patients as core value stakeholders,this program introduced a self-service examination appointment plat-form,established a follow-up and pre-visit process,and implemented time-slot-based appointments for diagnosis and treatment.These steps aim to promote the equitable allocation of medical resources and refine the follow-up and pre-visit model in public hospitals.Results Implementation at Hospital P demonstrated that after process reengineering,patients'average in-hospital time reduced by nearly 135 minutes,significantly enhancing treatment efficiency,satisfaction levels among both medical staff and patients,and accessibility of medical services.Conclusion Grounded in value co-creation theory,the follow-up and pre-visit model of Hospital P is divided into three stages:mutual value positioning of both doctors and patients,collaborative value crea-tion,and sustained value continuation for both.This alignment between process reengineering goals and value co-creation out-comes helps improve the quality of medical services,better meet the medical needs of patients,and achieve a win-win scenario for both doctors and patients.

Objective:To study the surface characterization and bonding strength of zirconia ceramic following nonthermal argon plasma(NTAP)treatment for different times.Methods:Zirconia ceramics were cut into 2 sizes of specimens(Ⅰ:10 mm×10 mm ×2 mm,Ⅱ:3 mm3 mm×2 mm),which were respectively subdivided into 5 groups:blank control group(A),sandblasting group(B)and NTAP treatment for 60,90 and 120 s groups(C,D and E respectively).The changes in surface morphology,roughness,C and O elements and contact angle of the sample surfaces were tested with Type Ⅰ specimens.Type Ⅱ specimens were applied to the uniform surface treatments and were cemented to isolated teeth using the RelyX U200 cement,the samples were submitted to a shear bond strength(SBS)test.Results:SEM and AFM show that NTAP treatment does not alter the surface morphology and roughness of zirconia ceramics.C element decreased and O element increased,and the contact angle became smaller after NTAP treatment.Among the 5 groups,group D showed the most extensive changes.And the result of SBS was as follows of NTAP treat-ment time:0 s＜60 s＜90 s=120 s(among 0,60 and 90 s groups,P＜0.05).Conclusion:NTAP treatment can improve the bond strength of zirconia ceramic by increasing its surface energy and wettability,and NTAP treatment for 90 s may be a suitable surface treatment method for zirconia ceramics.

Objective:To investigate the effect of astragalus polysaccharide(APS)on macrophage phagocytosis of P.aerugino-sa and its related mechanism.Methods:The effect of APS on the activity of macrophages was detected by CCK-8 assay.The effect of APS on macrophage phagocytosis related signal pathways were detected by Western blot.Gentamicin protection test and flow cytometry were employed to evaluate the effect of APS on macrophage phagocytosis of P.aeruginosa.Then the macrophages were pretreated with the inhibitors of TLR4,NF-κB,STAT3,MAPK and PI3K to assess the mechanism mediating APS on macrophage phagocytosis.Results:APS promotes the activity of macrophages,and significantly promoted the phagocytosis of P.aeruginosa by macrophages.Fur-thermore,macrophage phagocytosis of P.aeruginosa was significantly inhibited by TLR4 inhibitors(TAK242)and PI3K inhibitors(LY294002),but not affected by NF-κB,STAT3 and MAPK inhibitors.Conclusion:APS has no toxicity on macrophages,and can promote the phagocytosis of macrophages to P.aeruginosa,the underlying mechanism may be related to TLR4/PI3K pathway.

Objective:In view of the incomplete evaluation system of actual combat injury treatment ability of grassroots officers and soldiers at present, this paper aims to construct a multi-dimensional evaluation system for combat injury treatment to provide theoretical guidance and evaluation means for warfighting-oriented training on combat injury treatment.Methods:Through literature review, expert consultation, and hierarchical analysis, we determined the factors to evaluate the comprehensive ability of field first aid for combat injuries of grassroots officers and soldiers from the perspective of actual combat, and then established a system for evaluating the actual combat injury treatment ability of grassroots officers and soldiers, in which the weights of factors in each dimension were determined.Results:An evaluation system of actual combat injury treatment ability of grassroots officers and soldiers was formed, which included 4 first-level indices and 16 second-level indices in tactics, strategies, treatment, and protection dimensions.Conclusions:The established evaluation system can provide a reference for actual combat injury treatment training at the grassroots level.