ObjectiveTo validate the therapeutic effect of Yinqi Sanhuang Jiedu decoction （YQSH） on a mouse model of carbon tetrachloride （CCl₄）-induced liver fibrosis and to investigate its correlations with the Piezo-type mechanosensitive ion channel component 1 （Piezo1）-Yes-associated protein （YAP） mechanical signaling axis and macrophage polarization. MethodsFifty-four C57BL/6J mice were randomized into a blank control group， a 4-week model group， a 6-week model group， a 8-week model group， a positive drug （silymarin， 55 mg·kg^-1·d^-1） group， and low-， medium-， and high-dose （8.325， 16.65， 33.3 g·kg^-1·d^-1， respectively） YQSH groups. Except the 6-week model group （n=12）， each of the other groups had 6 mice. Mice in other groups except the blank control group received intraperitoneal injections of 10% CCl₄ twice weekly for the modeling of liver fibrosis. Drug interventions began one week after the initial modeling through gavage， and the blank control and model groups received 0.2 mL of normal saline via gavage. The histopathological changes and collagen deposition in the liver were observed via hematoxylin-eosin （HE）， Masson's trichrome， and Sirius red staining. Serum activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， as well as serum levels of total protein （TP）， albumin （ALB）， total bilirubin （TBIL）， hyaluronic acid （HA）， laminin （LN）， procollagen type Ⅲ （PCⅢ）， and collagen type Ⅳ （Ⅳ-C）， were measured. The levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA levels of Piezo1 and YAP1 in the liver tissue were determined by immunohistochemistry （IHC） and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， respectively. Co-localization of Piezo1 with YAP1， and YAP1 with inducible nitric oxide synthase （iNOS） was observed by the immunofluorescence （IF） assay. The proportions of M1-type （F4/80⁺CD80⁺） and M2-type （F4/80⁺CD206⁺） macrophages in the liver tissue were examined by flow cytometry. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses and gene set enrichment analysis （GSEA） were performed through transcriptomic sequencing. ResultsCompared with the blank control group， the model group exhibited gradually worsened liver fibrosis at the 4^th， 6^th， and 8^th weeks. The 6- and 8-week model groups showcased inflammatory cell infiltration and collagen deposition in the liver （P<0.01） and upregulated protein levels of both Piezo1 and YAP1 （P<0.01）. Compared with the model groups， treatment with YQSH improved the liver function （P<0.05， P<0.01）， alleviated liver fibrosis （P<0.05， P<0.01）， and reduced intrahepatic inflammatory cell infiltration and collagen deposition （P<0.01）. Furthermore， the treatment downregulated the protein and mRNA levels of Piezo1 and YAP1 （P<0.05， P<0.01）， lowered the levels of inflammatory factors TNF-α and IL-1β （P<0.05， P<0.01）， and decreased the ratio of CD80 （M1-type）/CD206 （M2-type） macrophage proteins （P<0.01）. The IF assay showed co-localization of YAP1 with Piezo1 and iNOS. Compared with the model groups， YQSH treatment downregulated the expression of Piezo1， YAP1， and iNOS （P<0.05， P<0.01）. Transcriptomic analysis suggested that the anti-liver fibrosis effect of YQSH may be related to the Hippo signaling pathway （P<0.05）. ConclusionThe anti-liver fibrosis effect of YQSH may be related to its inhibition of the Piezo1-YAP signaling axis and regulation of macrophage polarization.

ObjectiveTo validate the therapeutic effect of Yinqi Sanhuang Jiedu decoction （YQSH） on a mouse model of carbon tetrachloride （CCl₄）-induced liver fibrosis and to investigate its correlations with the Piezo-type mechanosensitive ion channel component 1 （Piezo1）-Yes-associated protein （YAP） mechanical signaling axis and macrophage polarization. MethodsFifty-four C57BL/6J mice were randomized into a blank control group， a 4-week model group， a 6-week model group， a 8-week model group， a positive drug （silymarin， 55 mg·kg^-1·d^-1） group， and low-， medium-， and high-dose （8.325， 16.65， 33.3 g·kg^-1·d^-1， respectively） YQSH groups. Except the 6-week model group （n=12）， each of the other groups had 6 mice. Mice in other groups except the blank control group received intraperitoneal injections of 10% CCl₄ twice weekly for the modeling of liver fibrosis. Drug interventions began one week after the initial modeling through gavage， and the blank control and model groups received 0.2 mL of normal saline via gavage. The histopathological changes and collagen deposition in the liver were observed via hematoxylin-eosin （HE）， Masson's trichrome， and Sirius red staining. Serum activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， as well as serum levels of total protein （TP）， albumin （ALB）， total bilirubin （TBIL）， hyaluronic acid （HA）， laminin （LN）， procollagen type Ⅲ （PCⅢ）， and collagen type Ⅳ （Ⅳ-C）， were measured. The levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA levels of Piezo1 and YAP1 in the liver tissue were determined by immunohistochemistry （IHC） and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， respectively. Co-localization of Piezo1 with YAP1， and YAP1 with inducible nitric oxide synthase （iNOS） was observed by the immunofluorescence （IF） assay. The proportions of M1-type （F4/80⁺CD80⁺） and M2-type （F4/80⁺CD206⁺） macrophages in the liver tissue were examined by flow cytometry. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses and gene set enrichment analysis （GSEA） were performed through transcriptomic sequencing. ResultsCompared with the blank control group， the model group exhibited gradually worsened liver fibrosis at the 4^th， 6^th， and 8^th weeks. The 6- and 8-week model groups showcased inflammatory cell infiltration and collagen deposition in the liver （P<0.01） and upregulated protein levels of both Piezo1 and YAP1 （P<0.01）. Compared with the model groups， treatment with YQSH improved the liver function （P<0.05， P<0.01）， alleviated liver fibrosis （P<0.05， P<0.01）， and reduced intrahepatic inflammatory cell infiltration and collagen deposition （P<0.01）. Furthermore， the treatment downregulated the protein and mRNA levels of Piezo1 and YAP1 （P<0.05， P<0.01）， lowered the levels of inflammatory factors TNF-α and IL-1β （P<0.05， P<0.01）， and decreased the ratio of CD80 （M1-type）/CD206 （M2-type） macrophage proteins （P<0.01）. The IF assay showed co-localization of YAP1 with Piezo1 and iNOS. Compared with the model groups， YQSH treatment downregulated the expression of Piezo1， YAP1， and iNOS （P<0.05， P<0.01）. Transcriptomic analysis suggested that the anti-liver fibrosis effect of YQSH may be related to the Hippo signaling pathway （P<0.05）. ConclusionThe anti-liver fibrosis effect of YQSH may be related to its inhibition of the Piezo1-YAP signaling axis and regulation of macrophage polarization.

Objective To analyze the psychological status and related factors in elderly patients of different genders after percutaneous coronary intervention(PCI)for coronary heart disease(CHD).Methods A total of 303 eligible patients treated in our department from January 2021 to December 2024 were enrolled,and then divided into a male group(154 cases)and a female group(149 cases).Based on the 7-item Generalized Anxiety Disorder Scale,they were assigned into a female anxiety group(97 cases),a female non-anxiety group(52 cases),a male anxiety group(82 cases)and a male non-anxiety group(72 cases).According to the 9-item Patient Health Questionnaire,they were alsocategorized into a female depression group(92 cases),a female non-depression group(57 cases),a male depression group(75 cases)and a male non-depression group(79 cases).The basic clinical data and results of psychological assessment were recorded in each group.Statistical analysis was performed on the indicators,including total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C).Results The female patients had significantly higher scores of physical symptoms,anxiety and depression,and larger proportions of anxiety and depression than the male group(P＜0.05,P＜0.01).In the female patients,advance age,higher TC,TG and LDL-C levels and somatic symptom score were observed in those with anxiety than those without,and in those with depression group than those without(P＜0.05,P＜0.01).The male anxiety group exhibited older age,more lesion vessels,larger ratio of ≥2 stents,and higher physical symptom score when compared with the male non-anxiety group,and so were in the male depression group than the male non-depression group(P＜0.05,P＜0.01).Binary logistic regression analysis showed that age,TC,LDL-C,and somatic symptom score were risk factors for anxiety,and age,TC,HDL-C,LDL-C,and somatic symptom score were risk factors for depression in elderly female CHD patients(P＜0.05,P＜0.01).For the elderly male CHD patients,age,stents ≥2,and somatic symptom score were risk factors for anxiety(OR=1.068,95％CI:1.002-1.138,P＜0.05;OR=6.458,95％CI:2.807-14.858,P＜0.01;OR=1.148,95％CI:1.045-1.262,P＜0.01),and age,stents ≥2,and more lesion vessels were risk factors for depression(P＜0.01).Conclusion The female CHD patients undergoing PCI have higher anxiety and depression scores than the male patients.There are certain differences in the influencing factors for anxiety and depression between genders.Effective control of these risk factors may help improve the prognosis of CHD patients after PCI.

Objective To analyze the psychological status and related factors in elderly patients of different genders after percutaneous coronary intervention(PCI)for coronary heart disease(CHD).Methods A total of 303 eligible patients treated in our department from January 2021 to December 2024 were enrolled,and then divided into a male group(154 cases)and a female group(149 cases).Based on the 7-item Generalized Anxiety Disorder Scale,they were assigned into a female anxiety group(97 cases),a female non-anxiety group(52 cases),a male anxiety group(82 cases)and a male non-anxiety group(72 cases).According to the 9-item Patient Health Questionnaire,they were alsocategorized into a female depression group(92 cases),a female non-depression group(57 cases),a male depression group(75 cases)and a male non-depression group(79 cases).The basic clinical data and results of psychological assessment were recorded in each group.Statistical analysis was performed on the indicators,including total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C).Results The female patients had significantly higher scores of physical symptoms,anxiety and depression,and larger proportions of anxiety and depression than the male group(P＜0.05,P＜0.01).In the female patients,advance age,higher TC,TG and LDL-C levels and somatic symptom score were observed in those with anxiety than those without,and in those with depression group than those without(P＜0.05,P＜0.01).The male anxiety group exhibited older age,more lesion vessels,larger ratio of ≥2 stents,and higher physical symptom score when compared with the male non-anxiety group,and so were in the male depression group than the male non-depression group(P＜0.05,P＜0.01).Binary logistic regression analysis showed that age,TC,LDL-C,and somatic symptom score were risk factors for anxiety,and age,TC,HDL-C,LDL-C,and somatic symptom score were risk factors for depression in elderly female CHD patients(P＜0.05,P＜0.01).For the elderly male CHD patients,age,stents ≥2,and somatic symptom score were risk factors for anxiety(OR=1.068,95％CI:1.002-1.138,P＜0.05;OR=6.458,95％CI:2.807-14.858,P＜0.01;OR=1.148,95％CI:1.045-1.262,P＜0.01),and age,stents ≥2,and more lesion vessels were risk factors for depression(P＜0.01).Conclusion The female CHD patients undergoing PCI have higher anxiety and depression scores than the male patients.There are certain differences in the influencing factors for anxiety and depression between genders.Effective control of these risk factors may help improve the prognosis of CHD patients after PCI.

Background Retinal vein occlusion is a common retinal vascular diseases.Thromblysis and anticoagulation therapies are main approaches.However,systemic thrombolysis is relatively inefficient,and it often enhances the risk of hemorrhage.Objective This study was to investigate the therapeutic effects of PLM-ΔK,a kringle deficiency mutant of plasmin,on photochemically induced branch retinal vein occlusion (BRVO) after intravitreal injection.Methods BRVO models were established by the combination of caudal vein injection of Rose Bengal with argon laser radiation of periphery area of retinal veins in SD rats.Forty model rats were randomized into balance salt solution (BSS) group and 0.01 U,0.02 U,0.03 U PLM-ΔK group,and 10 μl corresponding drug was intravtreally injected 12 hours after modeling.Ophthalmoscopy and fundus fluorescein angiography (FFA) were performed to observe the change of retinal veins.The animals were sacrificed 3 days after intravitreal injection,and hematoxylin and eosin staining was used for the histopathological and ultrastructural examination of retinas.The retina of the rats was isolated for the stretched preparation of retina.The expressions of fibronectin (FN) and laminin (LN) in eyeball wall were assayed by immunofluorescence technology.The use and care of the animals complied with Statement of the Association for Research in Vision and Ophthalmology.Results The revascularization of over 2 retinal veins was found in 0,3,6 and 8 rats in the BBS group and 0.01 U,0.02 U,0.03 U PLM-ΔK group 3 days after intravitreal injection,respectively,showing a significant difference among the groups (x2=9.635,P =0.022),and the rat number with revascularization in 0.01 U PLM-ΔK group was not significantly different from that in BSS group (Z=-1.558,P =0.119),but the difference between 0.03 U PLM-ΔK group and 0.01 U PLM-ΔK group was significant (Z=-2.762,P=0.006).In the third day after intravitreal injection,retinal vein thrombus were found in the BSS group under the light microscope,and angiogenesis was seen on the retinal flatmount nuclear.In the 0.03 U PLM-ΔK group,posterior vitreal detachment was exhibited under the light microcope,and no retinal new vessel and cell damage were seen.FN was strongly expressed in the inner limiting membrane (ILM) layer,photocyte layer,outer limiting membrane (OLM) layer,choroid and scleral layer,and LN was expressed mainly in the ILM,OLM and scleral layer in the BSS group.However,the expression intensities of FN and LN were obviously weakened in the 0.03 U PLM-ΔK group.Conclusions Intravitreal injection of PLM-ΔK can enhance the reperfusion of occluded branch retinal vein and serve as a potential therapeutic drug for BRVO.Also it can permeate into choroid after intravitreal injection to degradate FN and LN.

Objective To study the effect of retinal dehydrogenase type 2 inhibitor (4-diethylaminobenzaldehyde,DEAB) on embryonic CSrdiac develclpment of zebrafish model with retinoic acid(RA)deficiency. Methods Zebrafish embryos were treated with DEAB at various concentrations including 1×10~(-6),5×10~(-6),10×10~(-6),25×10~(-6)mol/L at 5,8 and 10.3 hours post fertilization,respectively.The effects of DEAB on the embryonic development were assessed under microscope.1×10~(-9)mol/L exogenous RA was then added to detect the antagonistic effect against DEAB.The abnormal cardiac phenotype,heart rate and ventricular systolic fraction were observed and analyzed between wild type and DEAB treated groups.The expression of specific cardiac gene, natriuretic peptide precursor A,was monitored by whole-mount in situ hybridization to demonstrate the effect of RA signaling on early cardiac development. Results The survival rate of zebrafish embryos declined with the increase of DEAB concentration at different developmental stage.The percentage of abnormal embryos reached 100% when DEAB over 5×10~(-6)mol/L.1×10~(-9) mol/L exogenous RA could eliminate the teratogenic effect of DEAB(≥5×10~(-6)mol/L).DEAB treated embryos presented abnormal cardiac phenotype,including tubular heart,incomplete D-loop,abnormal atrioventricular development,regurgitation,slow blood flow and weak heart beat.The difference of heart rate and ventrieular systolic fraction between wild type and RA deficiency embryos was of statistical significance(P<0.05).The natriuretic peptide precursor A expression remained in the ventricle,but reduced obviously in the atrium with RA signaling deficiency. Conclusions The effects of DEAB on the embryonic development are dose-dependent and time-dependent,and could be rescued by exogenous RA.RA signaling plays a critical role in several key stages of early cardiac development and natriuretie peptide precursor A expression.

Whole mount in situ hybridization with BHC80 RNA probe showed that BHC80 was expressed in zebrafish central nervous system. Morpholino-modified antisense oligonucleotide was injected into zebrafish zygotes to knock down BHC80 expression. BHC80 knockdown resulted in striking decrease of erythrocytes and accumulation of erythrocytes at PBI. Further investigation of embryonic erythrocytes marker βe3 globin and hematopoiesis transcription factors gata1, c-myb and lmo2 by in situ hybridization showed that the erythroid progenitors marked with gata1 in BHC80 knockdown embryos were high proliferation and their differentiation were delayed, which led to decrease of erythrocytes and accumulation of erythrocytes at PBI. Both in situ hybridization and microangiography indicated that vasculature pattern of BHC80 knockdown embryos were almost normal.

Purpose The main purposes of our research were to: 1. set up the method of the RGDHirudin microsphere preparation; 2. set up the method to test the activity and the content of the medicine contained in the microsphere; 3. analyse the key factors on the quality of the microsphere preparation. Methods Co-poly lactic acid glycolic acid (PLGA) microsphere was prepared by a modified solvent evaporation method by a double emulsion with the use of polyvinylalcohol (PVA) as emulsification; PLGA was used as biodegradable material and dichloromethane as organic solvent. The influence of formulation factors including the W1/O on microsphere diameter distribution and yield coefficient;PVA concentration on microsphere appearance, encapsulation and yield coefficient; ultrasound on spherulization average and medicine activity; stirring speed on spherulization average and microsphere appearance; PLGA on microsphere appearance and microsphere dispersity; concentration of NaCl on encapsulation efficiency, yield coefficient and medicine content etc were studied. Results The size of all the fabricated microsphere was measured according to the several factors that affect the particle size. The average diameter was 81.38 μm, which is good for further research. The medicine content and the percent yield of all the microsphere was high, which ranged from 83. 92% - 96. 3% and 79.93% - 95.05% respectively. The encapsulation efficiency was about 23.95% - 65. 13%. We found that the concentration of the NaCl and PVA were the very important factors to the encapsulation efficiency. Physiological activity of RGD-Hirudin containing in the microsphere and the release rate of the microsphere were controlled. Furthermore, the release rate was stable. Conclusions The physiologic activity of RGD-Hirudin released from the microspheres was stable. PLGA-RGD-Hirudin microspheres were controlled released by the in vitro studies. Therefore, the in vivo experiment was well grounded.

AIM: Endothelial progenitor cells(EPCs) are a group of stem cells/progenitor cells,which exist in postnatal body and can be of specially homing to the foci of angiogenesis and then differentiate into endothelial cells.This investigation was to study the method for culturing endothelia progenitor cells(EPCs) in vitro,and to observe its feasibility and condition formed vessel-like structure.METHODS: The cells were isolated from born marrow,peripheral blood,umbilical cord blood or spleen in different laboratories.The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells under the induction of special cytokines.The expression of CD34,VEGFR-2,AC133 and VE-cadherin were detected by fluorescence-activated cell sorting.The endothelial cell lineage was confirmed by DiI-ac-LDL up-taking and Ⅷ factor immunocytochemistry.RESULTS: The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro successfully,forming cord-like and tube-like structure.The EPCs derived from rabbit peripheral blood differentiated more mature and formed vessel-like structure.CONCLUSION: The EPCs derived from human umbilical cord blood and rabbit peripheral blood formed vessel-like structure in vitro.EPCs may be a potential resource of vessel tissue engineering.