Objective To investigate the histological and cellular bases for the improvement of portal hypertension(PH)by observing liver histopathological changes after treatment in patients with cirrhotic portal hypertension,and to provide a basis for clinical drug development.Methods A total of 322 patients with hepatitis B cirrhosis who completed 48 weeks of antiviral therapy or combined anti-fibrotic treatment in 20 hospitals across 12 provinces in China from September 2014 to October 2018 were enrolled,and the noninvasive diagnostic criteria for clinically significant portal hypertension(CSPH)from Baveno Ⅶ were used to assess the severity of PH;43 patients with a confirmed diagnosis of CSPH were identified based on liver stiffness measurement(LSM)≥25 kPa before treatment,and according to whether the severity of PH was reduced by≥2 grades after treatment,the patients were divided into PH improvement(n=19)group and PH non-improvement group(n=24).Related data were collected,including demographic data,laboratory tests.Liver fibrosis were assessed,including HE staining and reticular fiber staining;liver microvascular lesions were assessed,including obliterative portal venopathy(OPV),nodular regenerative hyperplasia(NRH),and incomplete septal fibrosis(ISF).Single immunohistochemical staining was performed for von Willebrand factor(vWF),and fibronectin;multiplex immunohistochemical staining was performed for fibrinogen,CD32b,CD31,alpha-smooth muscle actin(α-SMA).The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between two groups.Results After 48 weeks of treatment,43 patients had significant improvements in red blood cell count,alanine aminotransferase,aspartate aminotransferase,aspartate aminotransferase-to-platelet ratio index score,liver fibrosis grade,and PH grade(all P<0.05),among whom 19 patients showed a reduction in PH severity by≥2 grades(PH improvement group),while the remaining patients were enrolled as the PH non-improvement group.There was no significant difference in the outcome of liver fibrosis between the two groups(χ2=3.380,P=0.066).Microvascular lesion assessment showed that compared with the PH non-improvement group,the PH improvement group had significantly lower OPV severity,microvascular density(the expression level of vWF),and expression of fibronectin(all P<0.05),while there were no significant differences in NRH severity,ISF severity,and the expression level of fibrinogen between the two groups(all P>0.05).Cytological evaluation showed no significant differences in the expression levels of CD32b,CD31,and α-SMA between the two groups before and after treatment(all P>0.05),and comparison of the expression levels before and after treatment showed that the PH improvement group had a significant increase in the expression level of CD32b(t=-2.007,P=0.045)and a significant reduction in the expression level of α-SMA(t=2.628,P=0.013).Conclusion The pathological features of PH improvement are associated with liver fibrosis regression and the improvement in liver microvascular lesions,and at the cellular level,PH improvement is associated with the dedifferentiation of liver sinusoidal endothelial cells and the activated phenotype of hepatic stellate cells.

Objective:To investigate the glymphatic system dysfunction and white matter microstructural damage in type 2 diabetic mellitus patients with diabetic peripheral neuropathy (DPN), and to identify the early diagnostic imaging biomarkers.Methods:Thirty-one DPN patients and 31 type 2 diabetes mellitus (T2DM) patients who attended the First People's Hospital of Yancheng from March 2022 to October 2023 were included. In addition, 40 healthy controls (HC) were recruited. All subjects underwent 3.0T MRI scan with diffusion tensor imaging and 3D-T1WI sequences, and the diffusion tensor imaging analysis along the perivascular space(DTI-ALPS) index, perivascular space volume fraction in white matter(PVSVF-WM) and peak width of skeletonized mean diffusivity (PSMD) were calculated. SPSS 26.0 software was used to perform one-way ANOVA, t-tests, and Chi-square tests to compare clinical data and imaging indicators among the three groups. Partial correlation analysis was used to explore the relationships between DTI-ALPS index, PVSVF-WM, PSMD and clinical indicators in DPN patients. Finally, receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic value of DTI-ALPS index, PVSVF-WM, and PSMD for DPN. Results:The DPN group(1.46±0.19)had considerably lower DTI-ALPS index than the T2DM group (1.59±0.14) and HC group (1.60±0.17) (both P<0.05, Bonferroni correction). The DPN group (1.44±0.11) had significantly higher PVSVF-WM than both the T2DM group (1.35±0.14) and HC group (1.26±0.13) (both P<0.05, Bonferroni correction). The DPN group (1.84±0.31) and the T2DM group (1.83±0.25) had higher PSMD than HC group (1.60±0.24) (both P<0.05, Bonferroni correction), but the difference between the DPN group and the T2DM group was not statistically significant ( P>0.05). The DTI-ALPS index in the DPN group were negatively correlated with PSMD ( r=-0.379, P=0.035). The DTI-ALPS index was negatively correlated with the Toronto clinical scoring system (TCSS) score ( r=-0.456, P=0.01), while PSMD was positively correlated with TCSS scores ( r=0.686, P<0.001) in DPN group. The ROC curves showed that the area under the curve (AUC) was 0.777 ( P<0.001) for the combined diagnosis of DPN with the DTI-ALPS index, PVSVF-WM and PSMD. Conclusion:DPN patients exhibit glymphatic system dysfunction and white matter microstructural damage. The DTI-ALPS index and PSMD can serve as objective markers for assessing DPN severity. The combination of glymphatic system function indicators and white matter microstructural damage markers has moderate diagnostic value for peripheral neuropathy in T2DM patients.

Objective:To investigate the glymphatic system dysfunction and white matter microstructural damage in type 2 diabetic mellitus patients with diabetic peripheral neuropathy (DPN), and to identify the early diagnostic imaging biomarkers.Methods:Thirty-one DPN patients and 31 type 2 diabetes mellitus (T2DM) patients who attended the First People's Hospital of Yancheng from March 2022 to October 2023 were included. In addition, 40 healthy controls (HC) were recruited. All subjects underwent 3.0T MRI scan with diffusion tensor imaging and 3D-T1WI sequences, and the diffusion tensor imaging analysis along the perivascular space(DTI-ALPS) index, perivascular space volume fraction in white matter(PVSVF-WM) and peak width of skeletonized mean diffusivity (PSMD) were calculated. SPSS 26.0 software was used to perform one-way ANOVA, t-tests, and Chi-square tests to compare clinical data and imaging indicators among the three groups. Partial correlation analysis was used to explore the relationships between DTI-ALPS index, PVSVF-WM, PSMD and clinical indicators in DPN patients. Finally, receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic value of DTI-ALPS index, PVSVF-WM, and PSMD for DPN. Results:The DPN group(1.46±0.19)had considerably lower DTI-ALPS index than the T2DM group (1.59±0.14) and HC group (1.60±0.17) (both P<0.05, Bonferroni correction). The DPN group (1.44±0.11) had significantly higher PVSVF-WM than both the T2DM group (1.35±0.14) and HC group (1.26±0.13) (both P<0.05, Bonferroni correction). The DPN group (1.84±0.31) and the T2DM group (1.83±0.25) had higher PSMD than HC group (1.60±0.24) (both P<0.05, Bonferroni correction), but the difference between the DPN group and the T2DM group was not statistically significant ( P>0.05). The DTI-ALPS index in the DPN group were negatively correlated with PSMD ( r=-0.379, P=0.035). The DTI-ALPS index was negatively correlated with the Toronto clinical scoring system (TCSS) score ( r=-0.456, P=0.01), while PSMD was positively correlated with TCSS scores ( r=0.686, P<0.001) in DPN group. The ROC curves showed that the area under the curve (AUC) was 0.777 ( P<0.001) for the combined diagnosis of DPN with the DTI-ALPS index, PVSVF-WM and PSMD. Conclusion:DPN patients exhibit glymphatic system dysfunction and white matter microstructural damage. The DTI-ALPS index and PSMD can serve as objective markers for assessing DPN severity. The combination of glymphatic system function indicators and white matter microstructural damage markers has moderate diagnostic value for peripheral neuropathy in T2DM patients.

Objective To investigate the histological and cellular bases for the improvement of portal hypertension(PH)by observing liver histopathological changes after treatment in patients with cirrhotic portal hypertension,and to provide a basis for clinical drug development.Methods A total of 322 patients with hepatitis B cirrhosis who completed 48 weeks of antiviral therapy or combined anti-fibrotic treatment in 20 hospitals across 12 provinces in China from September 2014 to October 2018 were enrolled,and the noninvasive diagnostic criteria for clinically significant portal hypertension(CSPH)from Baveno Ⅶ were used to assess the severity of PH;43 patients with a confirmed diagnosis of CSPH were identified based on liver stiffness measurement(LSM)≥25 kPa before treatment,and according to whether the severity of PH was reduced by≥2 grades after treatment,the patients were divided into PH improvement(n=19)group and PH non-improvement group(n=24).Related data were collected,including demographic data,laboratory tests.Liver fibrosis were assessed,including HE staining and reticular fiber staining;liver microvascular lesions were assessed,including obliterative portal venopathy(OPV),nodular regenerative hyperplasia(NRH),and incomplete septal fibrosis(ISF).Single immunohistochemical staining was performed for von Willebrand factor(vWF),and fibronectin;multiplex immunohistochemical staining was performed for fibrinogen,CD32b,CD31,alpha-smooth muscle actin(α-SMA).The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between two groups.Results After 48 weeks of treatment,43 patients had significant improvements in red blood cell count,alanine aminotransferase,aspartate aminotransferase,aspartate aminotransferase-to-platelet ratio index score,liver fibrosis grade,and PH grade(all P<0.05),among whom 19 patients showed a reduction in PH severity by≥2 grades(PH improvement group),while the remaining patients were enrolled as the PH non-improvement group.There was no significant difference in the outcome of liver fibrosis between the two groups(χ2=3.380,P=0.066).Microvascular lesion assessment showed that compared with the PH non-improvement group,the PH improvement group had significantly lower OPV severity,microvascular density(the expression level of vWF),and expression of fibronectin(all P<0.05),while there were no significant differences in NRH severity,ISF severity,and the expression level of fibrinogen between the two groups(all P>0.05).Cytological evaluation showed no significant differences in the expression levels of CD32b,CD31,and α-SMA between the two groups before and after treatment(all P>0.05),and comparison of the expression levels before and after treatment showed that the PH improvement group had a significant increase in the expression level of CD32b(t=-2.007,P=0.045)and a significant reduction in the expression level of α-SMA(t=2.628,P=0.013).Conclusion The pathological features of PH improvement are associated with liver fibrosis regression and the improvement in liver microvascular lesions,and at the cellular level,PH improvement is associated with the dedifferentiation of liver sinusoidal endothelial cells and the activated phenotype of hepatic stellate cells.

BACKGROUND:Minipigs are often used in research fields such as skin injury,vascular trauma and cosmetic medicine because they are highly similar to human beings in terms of skin tissue structure and cardiovascular system.Hydrogel as a wound repair drug possesses a variety of excellent physicochemical properties such as strong water retention and adhesion,which can provide isolation moisturization and drug release for wounds. OBJECTIVE:To summarize and conclude the progress of the application of trauma models for different experimental purposes of hydrogel therapy for minipigs,to reveal the development status of various types of minipig trauma models,to analyze the deficiencies of minipig trauma models at the present stage. METHODS:The relevant articles published in Web of Science database and CNKI database from the establishment of each database to 2023 were checked,using"piglet,miniature pig,minipig,miniature pig;gel,hydrogel;trauma,injury,wound,lesion,incision"as Chinese search terms and"Miniature Swine,Miniature pig,minipig;gel,hydrogel;injury,wound,lesion,incision"as English search terms.A total of 438 Chinese and English documents were retrieved,and 59 documents were included in the study through the inclusion and exclusion criteria. RESULTS AND CONCLUSION:(1)At present,the main models used clinically for trauma repair are large animal species(dogs and pigs),rabbits,and rodents(rats and mice).Because the skin structure of the minipig is more like that of humans,the minipig is the most ideal animal model for trauma repair.(2)In the in-vitro skin injury model,skin defect model is the basic wound model,which can be divided into full skin defect model and medium-thickness skin defect model according to the depth of the wound defect.Burn wound model and infected wound model are multidimensional models with hot metal scald and bacterial culture imposed on the basis of the skin defect model,which have the advantages of high safety coefficient and low operation difficulty.(3)In the in-vivo trauma repair model,mini-pigs are used as esophageal cricothyrotomy model which is more in line with the pathological state of clinical diseases.Mini-pigs are used in the gastric perforation and vascular hemostasis model,which can visually demonstrate the stronger organ adhesion,hemostatic properties and tissue regeneration-promoting effects of the hydrogel.(4)The specific parts of the pig also has the corresponding mode of use:pig ear is usually used to evaluate the hydrogel drug delayed-release effect.Porcine cellular proteins and pig skin collagen are mostly used to prepare composite hydrogels of tissue origin.

Although significant progress has been made in the development of novel targeted drugs for the treatment of acute myeloid leukemia(AML)in recent years,chemotherapy still remains the mainstay of treatment and the overall survival is poor in most patients.Here,we demonstrated the antileukemia activity of a novel small molecular compound NL101,which is formed through the modification on bendamustine with a suberanilohydroxamic acid(SAHA)radical.NL101 suppresses the proliferation of myeloid malignancy cells and primary AML cells.It induces DNA damage and caspase 3-mediated apoptosis.A genome-wide clustered regularly interspaced short palindromic repeats(CRISPR)library screen revealed that phosphatase and tensin homologous(PTEN)gene is critical for the regulation of cell survival upon NL101 treatment.The knockout or inhibition of PTEN significantly reduced NL101-induced apoptosis in AML and myelodysplastic syndrome(MDS)cells,accompanied by the activation of protein kinase B(AKT)signaling pathway.The inhibition of mammalian target of rapamycin(mTOR)by rapamycin enhanced the sensitivity of AML cells to NL101-induced cell death.These findings uncover PTEN protein expression as a major determinant of chemosensitivity to NL101 and provide a novel strategy to treat AML with the combination of NL101 and rapamycin.

The macrophages,as a crucial component of the body's immune system,can be polarized into M1 and M2 types by different cellular molecules in various environments,and contribute to the progression of various diseases.In inflammatory responses,the M1 macrophages are primarily regarded as the pro-inflammatory cells,facilitate the inflammation progression,tissue destruction,and bone resorption,while M2 macrophages,as inflammatory cells,participate in tissue healing and bone repairment.In the tumor microenvironment,the roles of M1 and M2 macrophages are reversed.The periodontitis,pulpitis,and oral squamous cell carcinoma(OSCC)are the most prevalent inflammation and tumor in the oral cavity.Therefore,this article summarizes the relevant researches from home and abroad on the polarization of macrophages in oral inflammatory responses such as periodontitis,peri-implantitis,and pulpitis,bone remodeling during orthodontic treatment,and OSCC,and elucidate the metabolic activities of macrophages in infiammation,tumor,and bone remodeling and the mechanism of regulating the onset and development of diseases by the macrophage polarization,and provides new perspectives for the clinical treatment.

Objective To explore the safety and feasibility of the application of day surgery management model under enhanced recovery after surgery(ERAS)concept in patients undergoing gynecologic laparoscopic surgery.Methods A non-randomized concurrent control trial was conducted on the patients who underwent laparoscopic surgery in our department from January to August 2021.A total of 92 patients admitted on odd date were assigned into Ward B of our department and served as the control group,and another 96 patients hospitalized on even date were subjected into Ward A and served as the observation group.The control group was given the routine treatment schedule,including the relevant examinations after admission and general operation procedure during hospitalization.The observation group was under a day surgery management model based on the concept of ERAS,with aid of a day surgery team and optimized perioperative management measures,including pre-hospital rehabilitation,shortening water fasting before surgery,multi-mode analgesia,preventive antiemesis,intraoperative warmth,prevention of deep vein thrombosis,immediate postoperative eating and activity,and follow-up after discharge.Postoperative subjective comfort,intestinal function recovery,social and economic benefits,postoperative complications and inflammatory indicators were compared between the 2 groups.Results In 0～6,7～12 and 13～24 h after operation,the scores of thirst,hunger,nausea,pain,abdominal distension and pharyngeal discomfort were significantly lower in the observation group than the control group(P＜0.01).The observation group had obviously shorter length of hospital stay and ealier bowel sound recovery and first anal exhaust than the control group(P＜0.01).No postoperative complication,such as fall,unplanned secondary operation or wound infection was observed in both groups.The postoperative inflammatory indicators,including procalcitonin(PCT),neutrophil percentage(Neu％)and white blood cell count(WBC)were all in the normal ranges in the 2 groups at 24 h and 3 and 7 d after surgery.Statistical differences were found in firstly postoperative mobilization,length of hospital stay,hospitalization cost and patient satisfaction between the 2 groups(P＜0.01).Conclusion ERAS-based day surgery management model has the advantages of shortening hospital stay,reducing medical costs,promoting postoperative rehabilitation,and improving the comfort and satisfaction in patients undergoing gynecologic laparoscopic surgery.

This study aims to develop an improved cell screening system for farnesoid X receptor (FXR) agonists based on a dual luciferase reporter gene system. FXR response element (FXRE) fragments from FXR target genes were cloned and inserted into upstream of firefly luciferase (Luc) gene in the plasmid pGL4-luc2P-Hygro. In combination with the internal reference plasmid containing renilla luciferase, a dual luciferase reporter gene system was developed and used for high throughput screening of FXR agonists. After studying the effects of over-expression of RXR, mouse or human FXR, various FXRE fragments, and different ratio of FXR plasmid amount to reporter gene plasmid, induction efficiency of the screening system was optimized by the known FXR agonist GW4064, and Z factor for the system reached 0.83 under optimized conditions. In summary, an improved cell screening system based on double luciferase reporter gene detection system was developed to facilitate the discovery of FXR agonists, where a new enhanced FXRE element was formed by a superposition of multiple FXRE fragments from FXR target genes, instead of a superposition of traditional IR-1 (inverted repeats-1) fragments.
Humans ; Mice ; Animals ; Transcription Factors/genetics* ; DNA-Binding Proteins/genetics* ; Receptors, Cytoplasmic and Nuclear/genetics* ; Genes, Reporter ; Luciferases/genetics*

Objective To observe the characteristics of C57BL/6N-Tg(1.28HBV)/Vst transgenic hepatitis B virus(HBV-Tg)model mice and analyze their transcriptomic characteristics.Methods Twenty male HBV-Tg mice were divided into an experimental group and a wild-type(control)group(n = 10 mice per group).The virological characteristics of the model mice were evaluated according to serum levels of HBV DNA,HBsAg,and HBeAg,and expression levels of HBsAg and HBcAg in liver tissue.Serum levels of alanine transaminase(ALT)and aspartate transaminase(AST),hematoxylin and eosin(HE)and Sirius red staining,and hydroxyproline(Hyp)in liver tissue were detected to evaluate the degree of liver inflammation and fibrosis.Liver tissue samples were randomly selected from three mice in each group for RNA extraction for high-throughput transcriptome sequencing.Significantly differentially expressed genes were identified using R software.Functional enrichment of differential genes was determined by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses,and genes with significant differences were verified by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results ALT and AST levels were increased in the model group compared with the normal group,with the result for ALT being more significant(P<0.05).HE staining of liver tissue showed enlargement of the liver nucleus and swelling of some hepatocytes in the model group,while Sirius red staining showed a small amount of collagen deposition in the sink area and interlobule in the HBV transgenic group,in the shape of thin lines.A total of 1352 differential genes were obtained by screening(|logFC|>2 and P.adj<0.05),including 703 up-regulated and 649 down-regulated genes.KEGG analysis suggested that differential genes were mainly enriched in the peroxisome proliferator-activated receptor(PPAR)signaling pathway,retinol metabolism,fatty acid degradation,and other pathways(P.adj<0.05).The main significantly up-regulated genes included Cyp4a10,Cyp4a14,Acot1,Acot3,and Ehhadh,and the significantly down-regulated genes included Scn5a、Apol10b、Igddc4、Cxcl1、9530077C05Rik.The trend was consistent after RT-qPCR detection(P<0.05).Conclusions HBV-Tg mice have a tendency to develop spontaneous fibrosis.Transcriptomic analysis showed that chronic hepatitis B mainly involves PPAR signaling,retinol metabolism,fatty acid degradation,drug metabolism,and other pathways.