ObjectiveTo investigate the effect and mechanism of modified Sini San in ameliorating intestinal mucosal barrier by observing its effects on short chain fatty acids （SCFAs） and high mobility group protein B1 （HMGB1）/receptor of advanced glycation end products （RAGE） signaling pathways in chronic stress rats. MethodsThe 50 male SD rats were randomly divided into control group，model group，low-dose modified Sini San group （7.34 g·kg^-1·d^-1），high-dose modified Sini San group （14.68 g·kg^-1·d^-1），and Fructo-oligosaccharides group （3.15 g·kg^-1·d^-1），with 10 rats in each group. Except for the control group，all other groups were subjected to chronic unpredictable stress/social isolation to create a chronic stress model for 6 weeks. After 4 weeks of modeling，each treatment group was given corresponding drugs by gavage for 2 weeks while modeling. The control group and model group were given the same volume of physiological saline. The effects of Modified Sini San on behaviors，body weight，Bristol score in feces and fecal moisture content in chronic stress rats were observed. Hematoxylin and eosin （HE） staining was used to observe the pathological changes in the cecum. The content of SCFAs in the cecal contents of rats were detected by Gas chromatography-mass spectrometry （GC-MS）. Immunohistochemistry and Western blot were used to detect the expression of HMGB1/RAGE pathway related proteins in cecal tissue. The levels of ZO-1，Occludin，and Claudin-1 in the cecal tissue were detected by enzyme linked immunosorbent assay （ELISA）. ResultsCompared with the model group，the sucrose preference rate，total distance traveled and the number of grid crossings in the open field test of rats in the low-dose modified Sini San group were obviously increased （P<0.05， P<0.01），and the immobility time in the open field test and the immobility time in the forced swimming test of rats in the low-dose and high-dose modified Sini San groups were obviously reduced （P<0.05， P<0.01）. Meanwhile，the Bristol score and fecal moisture content of rats in the low and high dose groups of modified Sini San were obviously increased （P<0.05）. The low-dose group of modified Sini San had intact mucosal layer structure in the cecal tissue and reduced infiltration of inflammatory cells. The content of SCFAs in the cecal contents increased，with a obviously increase in the content of acetic acid，propionic acid，butyric acid，and isovaleric acid （P<0.05， P<0.01） and the expression levels of HMGB1，RAGE，Toll-like receptor 2（TLR2），Toll-like receptor 4（TLR4），tumor necrosis factor-α（TNF-α），and nuclear factor kappa-B p65（NF-κB p65） proteins in cecal tissue were significantly decreased （P<0.05， P<0.01） in low-dose group of modified Sini San. Meanwhile，the contents of ZO-1，Occludin，and Claudin-1 in the cecal tissue were obviously increased （P<0.01） in low-dose group of modified Sini San. ConclusionModified Sini San can improve the function of intestinal mucosal barrier in chronic stress rats by increasing the content of SCFAs in the intestine and inhibiting the HMGB1/RAGE pathway.

ObjectiveThis paper aims to investigate and evaluate the staged efficacy and safety of the representative empirical prescription of the “Zhu Ke Jiao” theory， Qijia Rougan prescription， combined with entecavir in the treatment of hepatic fibrosis in chronic hepatitis B. MethodsA multicenter randomized controlled clinical study was conducted， and 101 patients diagnosed with chronic hepatitis B-related hepatic fibrosis （CHB-HF） who met the diagnosis and inclusion criteria were randomly assigned to an observation group （Qijia Rougan prescription + entecavir） and a control group （entecavir）. The treatment duration was 24 weeks. Liver stiffness measurement （LSM）， fibrosis-4 index （FIB-4）， portal vein diameter， hepatitis B serology， biochemical indicators， hepatic fibrosis markers in serum ［hyaluronic acid （HA）， laminin （LN）， procollagen Ⅲ peptide （PⅢP）， and type Ⅳ collagen （Ⅳ-C）］， and traditional Chinese medicine syndrome scores were used as efficacy evaluation indicators. Efficacy assessments and explorations of different staged subgroups of Qijia Rougan prescription were conducted according to LSM values based on the Metavir pathological staging standard. ResultsA total of 98 cases were included for statistical analysis， with 49 cases in the observation group and 49 in the control group. The general data of the patients in both groups were comparable. Compared with the same group before treatment， the observation group showed a significant reduction in LSM and FIB-4 （P<0.01）， as well as notable improvements in LN， Ⅳ-C， and various TCM syndrome scores （P<0.05， P<0.01）. When compared to the control group after treatment， the observation group demonstrated significant improvements in LSM， FIB-4， and various TCM syndrome score indicators （P<0.05， P<0.01）， indicating that the observation group performed better than the control group. Subgroup analysis of the regression of hepatic fibrosis stages showed that compared to the same group before treatment， the observation group had better improvement in regression of stages F2 and F3 （P<0.05）. When compared to the control group after treatment， the observation group exhibited superior improvement in regression of stage F3 （P<0.05）. No adverse events occurred in either group during the treatment period. ConclusionCompared with entecavir alone， the combination of Qijia Rougan prescription and entecavir significantly improves the degree of hepatic fibrosis and clinical TCM symptoms in patients. The optimal intervention period is primarily during stage F3， which is a potential “interception” point of the “Zhu Ke Jiao” theory.

To investigate the effect of mesenchymal stromal cells (MSCs) on the fibrosis progression and endometrial regeneration in mouse models of endometrial injury, a mouse endometrial injury model was established by physical scraping combined with lipopolysaccharide (LPS) chemical induction, and the endometrium morphology and collagen deposition were evaluated by HE and Masson staining after unilateral administration of MSCs. RT-qPCR and immunofluorescence were used to detect the expression of epithelial markers Epcam and collagen I (Col1). Primary endometrial epithelial cells were isolated to assess p21 expression. Fibrosis-related genes and signaling pathways were validated using transcriptome sequencing and RT-qPCR. The experimental results showed that MSCs increased endometrial thickness and number of glands (P<0.05), up-regulated the expression of Epcam (P<0.01), while significantly reducing uterine collagen fiber deposition (P<0.05) and the expression of p21 in primary epithelial cells (P<0.0001). Transcriptomic analysis revealed that MSCs attenuated collagen deposition by modulating genes involved in the TGF-β signaling pathway. This study elucidates that MSCs facilitate uterine repair by reversing endometrial fibrosis and promoting epithelial regeneration, and their role is related to the regulation of the TGF-β/SMAD pathway and suppression of aberrant collagen accumulation.

Aim To investigate the role and mecha-nism of allopurinol(ALLO)in restoring the abnormal metabolism of kynurenine(Kyn)mediated by trypto-phan-2,3-dioxygenase 2(TDO2)to ameliorate ulcera-tive colitis(UC)in mice,and to provide experimental basis for the treatment of UC by ALLO.Methods A dextran sodium sulfate(DSS)-induced mouse UC mod-el was established,and the mice were randomly divided into the control group,the model group,the ALLO low,medium and high-dose groups(10,20,and 40 mg·kg-1),and the positive-significant salazosulfapyridine(SASP)(200 mg·kg-1)group.The body mass and disease activity index(DAI)scores of mice were re-corded;HE staining was performed to observe the de-gree of pathological damage in colon tissue;Western blot was performed to detect TDO2 protein expression in colon tissue;flow cytometry was performed to detect changes in the proportion of macrophages in spleen and mesenteric lymph nodes;ELISA was employed to de-tect the levels of TNF-α,IL-1β and IL-6 pro-inflamma-tory cytokines in the supernatant of colon tissue homog-enate;high performance liquid chromatography(HPLC)was used to detect the levels of tryptophan(Trp)and Kyn in the supernatant of colon tissue ho-mogenate.Results Compared with the model group,ALLO administration significantly ameliorated colonic histopathological injury in UC mice,decreased the pro-portion of macrophages in spleen and mesenteric lymph nodes,down-regulated the levels of TNF-α,IL-1 β,and IL-6 pro-inflammatory factors in serum of homogenate of colonic tissues,and inhibited the activity(Kyn/Trp ratio)and expression of TDO2 in colonic tissues.Con-clusion ALLO improves disease manifestations in mice with ulcerative colitis,which may be related to its restoration of abnormal Kyn metabolism.

Objective:To explore the coordinated development status of the medical service supply-economy-society trinity systems in Zhejiang Common Prosperity Demonstration Zone,and to offer references for formulating policies conducive to the efficient and coordinated development of these three systems.Methods:Based on the panel data from 2013 to 2022,this research was conducted on the 11 prefectural-level cities in Zhejiang Province.Firstly,an evaluation index system for the three systems was established,and the entropy method was employed to determine the weights of each index and calculate the comprehensive evaluation index.Secondly,a triangular model was introduced to delineate the relative relationships among the three systems,and the coupling coordination degree model was utilized to disclose the coordination degree of the three systems.Finally,the spatial autocorrelation analysis method was applied to investigate the spatial autocorrelation of the coupling coordination degree of the three systems.Results:(1)On the whole,the development status of the three systems has improved over time,yet the comprehensive development level remains to be improved.(2)Overall,each prefectural-level city has transited from a medical service supply-society-dominated development to a balanced and coordinated development of the medical service supply-economy-society trinity systems.The coupling coordination degree of the three systems has risen from 0.468(on the verge of imbalance)in 2013 to 0.609(primary coordination)in 2022,presenting an upward trend in general.However,the coordination level remains to be improved and there exist imbalances among regions.(3)The coupling coordination degree of the three systems in Zhejiang Province exhibits a significant positive spatial correlation,and the spatial distribution characteristics are relatively stable. Conclusion:The coupling coordination degree of the medical service supply-economy-society trinity systems in Zhejiang Province awaits further enhancement. At the level of Zhejiang Province,favorable policy support should be provided,the layout of medical resources should be rationally planned,and high-quality economic and social development should be promoted. Each prefectural-level city should formulate strategies for medical service supply,economic,and social development in accordance with its own development level and local conditions,strengthen inter-city linkage and cooperation,and thereby elevate the coordinated development level of the three systems.

The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

Oropharyngeal squamous cell carcinoma(OPSCC)is a malignant tumor originating from the squamous epithelium of the oro-pharyngeal mucosa,accounting for more than 90％of oropharyngeal malignancies.In recent years,human papillomavirus(HPV)infec-tion has become one of the primary etiological factors of oropharyngeal squamous carcinoma.The incidence of HPV-associated oropharyn-geal squamous carcinoma has been rising annually,with a noticeable trend toward younger populations,posing a significant threat to hu-man health.Due to the distinct biological behavior and clinical characteristics of HPV-associated oropharyngeal squamous carcinoma com-pared to its non-HPV-related counterpart,the diagnostic and treatment strategies for oropharyngeal squamous carcinoma have undergone substantial changes.Prevention and screening for oropharyngeal squamous carcinoma are of critical importance.The diagnostic and treat-ment process involves multi-disciplinary collaboration,including oral and maxillofacial surgery,otolaryngology,head and neck surgery,oncology,radiology and pathology.Based on evidence from clinical practice,a comprehensive,integrated diagnostic and therapeutic ap-proach has been established,centered around the concept of"prevention,screening,diagnosis,treatment,and rehabilitation",covering the entire patient lifecycle and providing a valuable reference for clinical practice.

Aim To investigate the role of tryptophan 2,3-dioxygenase(TDO2)in cisplatin-acute kidney in-jury(Cis-AKI)and to explore the mechanism of TDO2 in relation to apoptosis in tubular epithelial cells(TECs)to investigate the mechanism of TDO2 associ-ated with apoptosis.Methods An AKI model was es-tablished by intraperitoneal injection of cisplatin(Cis).Colorimetric assay was used to detect CRE and BUN levels,and PAS staining was employed to observe renal injury in mice.Immunohistochemistry was used to detect TDO2 protein expression and distribution and macrophage(F4/80+)infiltration;immunofluores-cence was used to detect the co-localization of TDO2 with the tubular marker LTL;TUNEL staining was used to detect apoptosis in mouse kidney;flow cytome-try was used to detect overexpression of human renal cortical proximal tubular epithelial cells(HK2)and apoptosis after administration of the TDO2 inhibitor 680C91;Western blot was used to detect TDO2 and NF-κB pathway protein levels in HK2 cells after over-expression and inhibition of TDO2.Results In the o-verall animal experiments,Cis-AKI mice showed signif-icantly higher levels of CRE and BUN and obvious tu-bular damage compared with the control group;at the same time,the renal tissues of Cis-AKI mice showed increased expression of F4/80,and the proportion of apoptotic cells in kidney cells was increased.Immuno-histochemistry and immunofluorescence showed that the expression of TDO2 increased,mainly localized in TECs.In cellular experiments,HK2 cells overexpress-ing TDO2 increased the proportion of apoptosis,and the expression of TDO2,p-IKBα,and p-p65 proteins was elevated,and p-IKBα/IκBα and p-p65/p65 were ele-vated;furthermore,the proportion of apoptosis was re-duced by the administration of 680C91,and the expres-sion of p-IκBα,and p-p65 proteins decreased,and the expression of p-IKBα/IKBα,and p-p65/p65 de-creased.Conclusions Elevated TDO2 in TECs is in-volved in the pathological mechanism of Cis-AKI,which may be related to its induction of apoptosis in TECs and activation of the NF-κB signaling pathway and consequently renal injury.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Aim To investigate the role and mecha-nism of allopurinol(ALLO)in restoring the abnormal metabolism of kynurenine(Kyn)mediated by trypto-phan-2,3-dioxygenase 2(TDO2)to ameliorate ulcera-tive colitis(UC)in mice,and to provide experimental basis for the treatment of UC by ALLO.Methods A dextran sodium sulfate(DSS)-induced mouse UC mod-el was established,and the mice were randomly divided into the control group,the model group,the ALLO low,medium and high-dose groups(10,20,and 40 mg·kg-1),and the positive-significant salazosulfapyridine(SASP)(200 mg·kg-1)group.The body mass and disease activity index(DAI)scores of mice were re-corded;HE staining was performed to observe the de-gree of pathological damage in colon tissue;Western blot was performed to detect TDO2 protein expression in colon tissue;flow cytometry was performed to detect changes in the proportion of macrophages in spleen and mesenteric lymph nodes;ELISA was employed to de-tect the levels of TNF-α,IL-1β and IL-6 pro-inflamma-tory cytokines in the supernatant of colon tissue homog-enate;high performance liquid chromatography(HPLC)was used to detect the levels of tryptophan(Trp)and Kyn in the supernatant of colon tissue ho-mogenate.Results Compared with the model group,ALLO administration significantly ameliorated colonic histopathological injury in UC mice,decreased the pro-portion of macrophages in spleen and mesenteric lymph nodes,down-regulated the levels of TNF-α,IL-1 β,and IL-6 pro-inflammatory factors in serum of homogenate of colonic tissues,and inhibited the activity(Kyn/Trp ratio)and expression of TDO2 in colonic tissues.Con-clusion ALLO improves disease manifestations in mice with ulcerative colitis,which may be related to its restoration of abnormal Kyn metabolism.