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.

Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

Objective To investigate the mechanism of liver-soothing and stomach-harmonizing recipe alleviating airway inflammation in rats with gastroesophageal reflux disease(GERD)by regula-ting peroxisome proliferator-activated receptor γ/retinoid X receptor(PPAR-γ/RXR)signaling path-way.Methods A rat model of GERD with airway inflammation was established by perfusion of hydro-chloric acid into the lower esophagus.A total of 50 SD male rats were randomly divided into control group,model group,low-dose liver-soothing and stomach-harmonizing recipe group(10.49 g/kg crude drug dosage),high-dose liver-soothing and stomach-harmonizing recipe group(20.98 g/kg crude drug dosage),and omeprazole group(3.67 mg/kg),with 10 rats in each group.The rats were gavaged for 14 days.Hematoxylin and eosin(HE)staining was used to observe the pathological change in tracheal tissue.RT-qPCR was used to detect the mRNA expression levels of inflammatory cytokines[interleukin(IL)-17,IL-33,inducible nitric oxide synthase(iNOS)]and anti-inflammatory cytokines[IL-10,Clara cell 16-kDa protein(CC16),surfactant protein-D(SP-D)]in bronchoalveolar lavage fluid;the Western blot was used to detect the relative protein expression levels of PPAR-γ,RXR-α,nuclear factor-KB(NF-κB),and activator protein-1(AP-1).Results The HE staining results showed that a large number of inflammatory cell infiltrations were observed in the model group,while the inflammatory cell infiltrations were significantly reduced in the low-dose liver-sooth-ing and stomach-harmonizing recipe group,high-dose liver-soothing and stomach-harmonizing recipe group,and omeprazole group.Compared with the control group,the mRNA expression levels of IL-17,IL-33 and iNOS in bronchoalveolar lavage fluid were significantly increased while the mRNA expression levels of IL-10,CC16 and SP-D were significantly decreased in the model group,and the relative protein expression levels of PPAR-γ and RXR-α were significantly increased,while the rela-tive protein expression levels of NF-κB and AP-1 were significantly decreased in the model group(P＜0.05).Compared with the model group,the above indicators were significantly improved in the high-dose liver-soothing and stomach-harmonizing recipe group and omeprazole group(P＜0.05).Conclusion Liver-soothing and stomach-harmonizing recipe can effectively alleviate airway inflam-mation in rats with GERD,and its mechanism may be related to the activation of the PPAR-y/RXR signaling pathway.

Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

Objective:To investigate corneal transparency alteration in patients with type 2 diabetes and its influencing factors.Methods:A case-control study was conducted.A total of 52 patients with type 2 diabetes mellitus (DM) (104 eyes) and 23 age-matched healthy controls (46 eyes) were enrolled as DM group and normal control group in the Second Affiliated Hospital of Anhui Medical University from October 1, 2020 to October 30, 2021.Patients with DM were further divided into non-diabetic retinopathy (non-DR) and DR groups according to their fundus conditions.Corneal densitometry (CD) was evaluated using the Pentacam.According to its built-in program, the cornea was divided into anterior, intermediate, and posterior layers and subdivided into 0-2 mm, >2-6 mm, >6-10 mm, and >10-12 mm annular regions with the corneal apex as the center of the circle.Pentacam automatically calculated the CD value of each corneal layer and region as well as the total CD value.The influencing factors of total CD value in diabetes group were analyzed by a multivariate linear regression analysis model.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Anhui Medical University (No.YX2023-129[F1]).Written informed consent was obtained from each subject before any medical examination.Results:The total CD value of diabetes group was 20.24±3.10, which was significantly higher than 18.79±3.31 of normal control group ( t=-2.583, P=0.011).The CD values of the anterior layer, intermediate layer, 0-2 mm, and >2-6 mm regions were significantly higher in diabetes group than in normal control group (all at P<0.05).The CD values in the anterior layer were higher in non-DR and DR groups than in normal control group, and the CD values and total CD values in the middle and posterior layers were higher in non-DR group than in normal control group and DR group, and the differences were statistically significant (all at P<0.05).The CD values in the 0-2 mm and >2-6 mm regions were significantly higher in non-DR group than in normal control group, and the CD value in the >6-10 mm annular region was significantly higher in non-DR group than in DR group and normal control group (all at P<0.05).Multivariate linear regression analysis showed that age and glycosylated hemoglobin (HbA1c) level were the main influencing factors for the increase in CD values in diabetic patients ( β=0.266, P<0.001; β=0.423, P=0.003). Conclusions:The decrease of corneal transparency precedes the appearance of DR in patients with diabetes.Poor control of HbA1c level in diabetic patients may cause the decline of corneal transparency.

ObjectiveTo investigate the effect of transplantation of bone marrow mesenchymal stem cells （BMSCs） co-cultured with bone marrow-derived M2 macrophages （M2-BMDMs）， named as BMSC^M2， on a rat model of liver cirrhosis induced by carbon tetrachloride （CCl₄）/2-acetaminofluorene （2-AAF）. MethodsRat BMDMs were isolated and polarized into M2 phenotype， and rat BMSCs were isolated and co-cultured with M2-BMDMs at the third generation to obtain BMSC^M2. The rats were given subcutaneous injection of CCl₄ for 6 weeks to establish a model of liver cirrhosis， and then they were randomly divided into model group （M group）， BMSC group， and BMSC^M2 group， with 6 rats in each group. A normal group （N group） with 6 rats was also established. Since week 7， the model rats were given 2-AAF by gavage in addition to the subcutaneous injection of CCl₄. Samples were collected at the end of week 10 to observe liver function， liver histopathology， and hydroxyproline （Hyp） content in liver tissue， as well as changes in the markers for hepatic stellate cells， hepatic progenitor cells， cholangiocytes， and hepatocytes. 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 N group， the M group had significant increases in the activities of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） （P<0.01）； compared with the M group， the BMSC and BMSC^M2 groups had significant reductions in ALT and AST （P<0.01）， and the BMSC^M2 group had significantly better activities than the BMSC group （P<0.05）. Compared with the N group， the M group had significant increases in Hyp content and the mRNA and protein expression levels of alpha-smooth muscle actin （α-SMA） in the liver （P<0.01）； compared with the M group， the BMSC and BMSC^M2 groups had significant reductions in Hyp content and the expression of α-SMA （P<0.05）， and the BMSC^M2 group had a significantly lower level of α-SMA than the BMSC group （P<0.01）. Compared with the N group， the M group had significant increases in the mRNA expression levels of the hepatic progenitor cell markers EpCam and Sox9 and the cholangiocyte markers CK7 and CK19 （P<0.01） and significant reductions in the expression levels of the hepatocyte markers HNF-4α and Alb （P<0.01）； compared with the M group， the BMSC and BMSC^M2 groups had significant reductions in the mRNA expression levels of EpCam， Sox9， CK7， and CK19 （P<0.05） and significant increases in the mRNA expression levels of HNF-4α and Alb （P<0.05）， and compared with the BMSC group， the BMSC^M2 group had significant reductions in the mRNA expression levels of EpCam and CK19 （P<0.05） and significant increase in the expression level of HNF-4α （P<0.05）. ConclusionM2-BMDMs can enhance the therapeutic effect of BMSCs on CCl₄/2-AAF-induced liver cirrhosis in rats， which provides new ideas for further improving the therapeutic effect of BMSCs on liver cirrhosis.

Objective To investigate the effect and mechanism of Yiguan Decoction(YGJD)on the efficacy of M1 bone marrow-derived macrophages(M1-BMDMs)in the treatment of rats with liver cirrhosis induced by 2-AAF/CCl4.Methods BMDMs were isolated and induced into M1-BMDMs by lipopolysaccharide.A total of 50 male Wistar rats were randomly divided into normal group with 5 rats and model group with 45 rats.The rats for modeling were given subcutaneous injection of 50%CCl4 twice a week.Since week 7,the rats for modeling were randomly divided into model group(M group),YGJD group,M1-BMDM group,M1-BMDM+YGJD group,and sorafenib(SORA)group,and they were given subcutaneous injection of 30%CCl4 to maintain the progression of liver cirrhosis and intragastric administration of 2-AAF.CCR2 inhibitors were added to the drinking water,and each group was given the corresponding intervention.Related samples were collected at week 9.The rats were observed in terms of serum liver function parameters,liver pathology,hydroxyproline(Hyp)content in liver tissue,hepatic stellate cell activation,hepatic fibrosis and inflammation factors,and the expression levels of molecules associated with the Wnt signaling pathway.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.Results Compared with the M group,the M1-BMDM+YGJD group had significant reductions in the serum levels of alanine aminotransferase,aspartate aminotransferase,and total bilirubin(TBil)(all P<0.05)and a significant increase in the content of albumin(Alb)(P<0.05),and compared with the M1-BMDM group,the M1-BMDM+YGJD group had a significant reduction in the serum level of TBil(P<0.05)and a significant increase in the serum level of Alb(P<0.05).Compared with the M1-BMDM group,the M1-BMDM+YGJD group had significant reductions in the expression levels of CD68 and TNF-α(P<0.05).Compared with the M1-BMDM group,the M1-BMDM+YGJD group had significant reductions in Hyp content and Sirius red positive area(P<0.05).As for the non-canonical Wnt signaling pathway molecules,compared with the M1-BMDM group,the M1-BMDM+YGJD group had significantly lower mRNA and protein expression levels of Wnt5a(P<0.05)and mRNA expression level of Fzd2(P<0.05),as well as significant reductions in the mRNA expression levels of Wnt4,Wnt5b,and Fzd3(P<0.05),while there were no significant changes in the mRNA expression levels of the canonical Wnt signaling pathway molecules β-catenin,LRP5,LRP6,Fzd5,and TCF.Conclusion YGJD can enhance the therapeutic effect of M1-BMDMs on rats with liver cirrhosis induced by 2-AAF/CCl4,possibly by inhibiting the non-canonical Wnt5a/Fzd2 signaling pathway,which provides new ideas for the synergistic effect of traditional Chinese medicine on M1-BMDMs in the treatment of liver cirrhosis.

Objective To investigate the current status and existing problems in the implementation of stroke care continuity in China, and to collect relevant suggestions for solving these problems. Methods Focus group interviews were used, to survey thirty-six nursing managers and senior nurses in neurology departments or rehabilitation programs in 24 cities of 13 provinces. The interview team members presided over the meetings, with full-time staff taking notes and recordings, and the results of the interviews were summarized and organized in a timely manner after the interview. Results At present, most of the general hospitals surveyed are doing their best to continue supporting nursing care for patients discharged from hospitals in different forms following their rescue of the acute phase. However, they are faced with such challenges as insufficient nursing manpower, and discontinuity between different medical institutions, which result in oversimplified nursing care and fragmented care. Conclusions Stroke nursing practitioners are working on continuous nursing care, with quite some challenges as well. Nursing care continuity is an important step towards establishing and improving a scientific and rational grading nursing care system.

Objective To investigate the expression and clinical significance of miR-126 and vascular endothelial growth factor (VEGF) in proliferative diabetic retinopathy (PDR).Methods 226 cases of diabetic retinopathy (DR) patients admitted in our hospital were studied,including 110 cases of PDR (group PDR),116 non proliferative diabetic retinopathy (NPDR) (group NPDR).80 patients with diabetes mellitus without retinopathy (NDR) were enrolled in DR group at the same period,another 80 healthy subjects (control group) were selected as control group.The plasma miR-126 level of all subjects was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR).The enzyme linked immunosorbent assay (ELISA) method was used to detect plasma VEGF level.The clinical diagnostic value of miR-126 and VEGF in PDR patients was further analyzed.Results Plasma levels of miR-126 in PDR group,NPDR group and NDR group were lower than those in control group (P ＜ 0.05),PDR group was lower than NPDR group and NDR group (P ＜ 0.05);plasma levels of VEGF in PDR group,NPDR group and NDR group were higher than those in control group (P ＜ 0.05),PDR group was higher than NPDR group and NDR group (P ＜ 0.05).Total cholesterol (TC),triglyceride (TG) and glycated hemoglobin (HbAlc) in PDR group were higher than those in NPDR group and NDR group (P ＜0.05),low density lipoprotein cholesterol (LDL-C) in PDR group,NPDR group and NDR group were higher than those in control group (P ＜ 0.05),and LDL-C in PDR group was higher than that in NPDR group and NDR group (P ＜0.05).High density lipoprotein cholesterol (HDL-C) in PDR group and NDR group was lower than that in control group (P ＜ 0.05),C-reactive protein (CRP) in PDR group,NPDR group and NDR group was higher than that in control group (P ＜ 0.05);plasma miR-126 levels in PDR group were negatively correlated with TC,LDL-C,CRP and HbA1c (P ＜ 0.05),but positively correlated with HDL-C (P ＜ 0.05),and no correlation with TG;the plasma levels of VEGF in PDR patients were positively correlated with TC,TG,LDL-C,CRP and HbAlc (P ＜0.05),but negatively correlated with the expressions of miR126 (r =-0.573,P =0.000);the AUC of miR-126 was 0.861,and when the cut-off value was ＜0.64,the diagnostic sensitivity and specificity were 82.50％,83.64％;the area under curve (AUC) of VEGF was 0.889,and when the cut-off value was ＜ 7.000,the sensitivity and specificity of diagnosis were 82.73％,86.25％;0.847 for the AUC of HbA1c,and the sensitivity and specificity 81.82％,87.50％ respectively.Conclusions Plasma miR-126 is low-expressed and VEGF is high-expressed in PDR patients,there is a negative correlation between the two indexes.They may be involved in the course of PDR through abnormal lipid metabolism and inflammatory reaction and may be a potential biomarker for early diagnosis of PDR.

Objective To explore the effect of fast track surgery concept(FTS) in the perioperative nursing of lung cancer after surgery by Da Vinci robot.Methods A total of 75 hospitalized patients diagnosed as lung cancer,underwent pulmonary lobectomy in our hospital were randomly divided into experimental group and control group.Patients in the experimental group were carried out rapid track surgery,while those in the control group were given routine nursing.Chest tube indwelling time,time to first exhaust,antibiotic use time,time of off-bed ambulance,hospitalization time,postoperative persistent pulmonary leakage,hypoxemia,pulmonary infection,atelectasis,arrhythmia and chylothorax were compared between the two groups.Results The recovery indicators of patients given FTS were better,while the incidence rate of complications was lower than that of the control group.Conclusion The concept of rapid rehabilitation surgery has a significant effect for patients with lung cancer resection,and it can significantly reduce the incidence of complications,and promote patients'postoperative recovery.