Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.

Objective This study aimed to systematically investigate the pharmacological substances and potential mechanisms of Zhenzhu Tongluo Wan in the treatment of peripheral nerve disorders.Methods Based on the TCMSP database and related literature,the active ingredients of Zhenzhu Tongluo Wan were determined.Target proteins for these active ingredients and those associated with peripheral neuropathy were predicted using the TCMSP,SwissTarget Prediction,GeneCard,OMIM,and DisGeNET databases.The intersection of these two sets of proteins was once considered a key target for the treatment of peripheral neuropathy.A protein-protein interaction(PPI)network of the intersection targets was constructed using the STRING database and analyzed using Cytoscape 3.9.1 software.Key targets obtained from the network were subjected to GO enrichment analysis and pathway annotation analysis using the DAVID database.The UHPLC-Q-Orbitrap HRMS technique was used to characterize the major chemical components of Zhenzhu Tongluo Wan.In addition,molecular docking,animal and cell experiments were performed to explore the mechanisms underlying the treatment of peripheral neuropathy with Zhenzhu Tongluo Wan.Results A total of 147 potential active ingredients and 333 intersection targets of Zhenzhu Tongluo Wan were screened.GO function and KEGG pathway analysis revealed that the major pathways involved include MAPK signaling pathway,NF-κB signaling pathway,PI3K-Akt signaling pathway,MAPK cascade and inflammatory response,among others.Based on high-resolution mass spectrometry analysis,33 major active ingredients were identified from Zhenzhu Tongluo Wan,focusing on those compounds with a Dgree value greater than the median.Molecular docking results further demonstrate good binding interactions between compounds such as luteolin,quercetin,tryptophan,arginine and kaempferol found inZhenzhu Tongluo Wan and key targets.Western blotting experiments showed that luteolin,quercetin and tryptophan could significantly inhibit the expression of MAPK8,P65 and TNF proteins in inflammatory cells.Animal experiments showed that Zhenzhu Tongluo Wan significantly improved hyperalgesia in rats with diabetic peripheral neuropathy,which may be related to the modulation of the expression of MAPK8,P65 and TNF proteins in the dorsal root ganglion.Conclusion Zhenzhu tongluo pills may play a role in the treatment of peripheral neuropathy by acting on the targets of MAPK8,P65 and TNF.This study lays a foundation for further research on the mechanism and pharmacodynamic substances of Zhenzhu Tongluo Wan in the treatment of peripheral neuropathy.

Objective This study aimed to systematically investigate the pharmacological substances and potential mechanisms of Zhenzhu Tongluo Wan in the treatment of peripheral nerve disorders.Methods Based on the TCMSP database and related literature,the active ingredients of Zhenzhu Tongluo Wan were determined.Target proteins for these active ingredients and those associated with peripheral neuropathy were predicted using the TCMSP,SwissTarget Prediction,GeneCard,OMIM,and DisGeNET databases.The intersection of these two sets of proteins was once considered a key target for the treatment of peripheral neuropathy.A protein-protein interaction(PPI)network of the intersection targets was constructed using the STRING database and analyzed using Cytoscape 3.9.1 software.Key targets obtained from the network were subjected to GO enrichment analysis and pathway annotation analysis using the DAVID database.The UHPLC-Q-Orbitrap HRMS technique was used to characterize the major chemical components of Zhenzhu Tongluo Wan.In addition,molecular docking,animal and cell experiments were performed to explore the mechanisms underlying the treatment of peripheral neuropathy with Zhenzhu Tongluo Wan.Results A total of 147 potential active ingredients and 333 intersection targets of Zhenzhu Tongluo Wan were screened.GO function and KEGG pathway analysis revealed that the major pathways involved include MAPK signaling pathway,NF-κB signaling pathway,PI3K-Akt signaling pathway,MAPK cascade and inflammatory response,among others.Based on high-resolution mass spectrometry analysis,33 major active ingredients were identified from Zhenzhu Tongluo Wan,focusing on those compounds with a Dgree value greater than the median.Molecular docking results further demonstrate good binding interactions between compounds such as luteolin,quercetin,tryptophan,arginine and kaempferol found inZhenzhu Tongluo Wan and key targets.Western blotting experiments showed that luteolin,quercetin and tryptophan could significantly inhibit the expression of MAPK8,P65 and TNF proteins in inflammatory cells.Animal experiments showed that Zhenzhu Tongluo Wan significantly improved hyperalgesia in rats with diabetic peripheral neuropathy,which may be related to the modulation of the expression of MAPK8,P65 and TNF proteins in the dorsal root ganglion.Conclusion Zhenzhu tongluo pills may play a role in the treatment of peripheral neuropathy by acting on the targets of MAPK8,P65 and TNF.This study lays a foundation for further research on the mechanism and pharmacodynamic substances of Zhenzhu Tongluo Wan in the treatment of peripheral neuropathy.

Objective:To investigate the association of polymorphisms at the matrix metalloproteinase(MMP)-3-1612 position of the promotor region with the inflammatory response and oxidative stress in elderly patients with cerebral ischemic stroke.Methods:In this retrospective study, 129 elderly patients with cerebral infarction diagnosed and treated in our hospital between March 2019 and March 2021 were enrolled as the study group, and 110 healthy subjects were selected as the control group.Polymorphisms of the MMP-3-1612 position in the promotor region, the inflammatory response and oxidative stress were examined using appropriate parameters and the associations between them were analyzed. Results:Compared with the control group, the proportions of patients with hypertension, diabetes, and smoking history in the study group were significantly higher( χ2=16.05, 17.19, 14.19, all P<0.05), and the levels of fasting blood glucose, low-density lipoprotein, and homocysteine were also significantly higher( t=6.22, 3.64, 2.69, all P<0.05).Meanwhile, compared with patients carrying the MMP-3-5A/6A or the MMP-3-6A/6A genotype, the levels of serum inflammatory markers such as high mobility group box-1 protein(HMGB1), fractalkine(FKN), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β)and interleukin-17(IL-17)in patients carrying the MMP-3 gene 5A/5A genotype were significantly higher(all P<0.05).In addition, the expression of serum oxidative stress-related molecules Kelch-like ECH-associated protein 1(Keap1), nuclear factor erythroid-2 related factor2(Nrf2), antioxidant response element(ARE), quinone oxidoreductase 1(NQO1), and heme oxygenase-1(HO-1)was also significantly increased(all P<0.05), but there was no difference in these markers between patients carrying the MMP-3-5A/6A genotype and patients carrying the MMP-3-6A/6A genotype( P>0.05).Patients carrying the 5A/5A genotype and the 6A/6A genotype exhibited only one 97 bp band and one 120 bp band, respectively, while the patients carrying the 5A/6A genotype exhibited two 97 bp bands and two 120 bp bands.There was no statistical difference in the number of patients carrying the 5A/6A genotype in the cerebral infarction group compared with the control group( P>0.05), and the number of patients carrying the 5A/5A genotype in the cerebral infarction group was higher than that in the control group(69% or 53.49% vs.35% or 31.82%, χ2=11.34, P<0.05).Polymorphisms of the MMP-3 gene had a positive correlation with the risk of stroke( r=0.25, P<0.05). MMP-3-1612 gene polymorphism( OR=7.21, 95% CI: 1.13-1.83, P=0.01), elevated blood glucose( OR=1.27, 95% CI: 1.18-2.06, P<0.001), high homocysteine( OR=1.05, 95% CI: 1.08-1.58, P<0.01), hypertension( OR=5.414, 95% CI: 1.140-4.46, P<0.01), elevated low-density lipoprotein( OR=4.03, 95% CI: 1.03-2.35, P=0.02), coronary heart disease( OR=1.17, 95% CI: 1.47-3.19, P<0.01)and diabetes( OR=8.52, 95% CI: 1.32-4.71, P<0.01)were risk factors for cerebral infarction. Conclusions:In elderly patients with cerebral infarction, polymorphisms of the MMP-3-1612 position in the promotor region is closely related to the risk of cerebral ischemic stroke, the inflammatory response and oxidative stress.MMP-3 gene polymorphisms are risk factors for stroke.

Abstract Astragalus polysaccharides(APS), as one of the main active components of Astragali Radix, a traditional famous tonic drug in China, has good pharmacological effects in regulating immunity and anti-oxidation. In recent years, with the success of PD-1/PD-L1 inhibitors and anti-CTLA-4 drugs in cancer treatment, immunotherapy has truly become one of the main clinical anti-tumor treatment methods. Aiming at the characteristics of tumor immune escape and tumor microenvironment change, this paper explores the clinical potential of APS as an anti-tumor adjuvant drug, and summarizes the role and mechanism of APS in inhibiting tumor immune escape, regulating tumor microenvironment, and improving immune function. To summarize and analyze the immune regulation of APS on various tumors in order to provide an objective basis for APS as a clinical anti-tumor adjuvant drug.

OBJECTIVE To establish a quantitative analysis method for the quality control components in Tenghuang jiangu capsules， and predict the possible action mechanism of the quality control components. METHODS Seven key quality control components in Tenghuang jiangu capsules were quantitatively analyzed by UHPLC-Q-Orbitrap HRMS. The “component-target” network was constructed based on network pharmacology， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and gene ontology （GO） function enrichment analysis were further conducted to find the key signaling pathways. RESULTS The average contents of succinic acid， hyperoside， gallic acid， kaempferol， naringin， naringenin and protocatechuic acid in 20 batches of Tenghuang jiangu capsules were 520.92， 67.67， 129.48， 4.74， 397.45， 5.66 and 376.62 μg/g， respectively. The results of network pharmacology showed that the 62 key target genes of the quality control components of the drug included AKT1， TNF， VEGFA， MMP9， PTGS2， etc. They were mainly enriched in cytokine receptor interaction， nuclear factor， tumor necrosis factor， interleukin 17， rheumatoid arthritis， Toll-like receptor and other signal pathways， involving inflammatory reaction， signal transduction， protein phosphorylation and other biological processes， kytoplasm， cell membrane and other cell components， as well as enzyme activity， energy activity and other molecular functions. CONCLUSIONS The established UHPLC- Q-Orbitrap HRMS method can be used for the quantitative analysis of the quality control components of Tenghuang jiangu capsule. Its quality control components may be mapped to inflammatory pathways related to bone diseases such as rheumatoid arthritis and Toll-like receptors through AKT1， TNF， VEGFA and other key targets， so as to play a therapeutic role.

Infectious diarrhea is an important public health problem, which has a significant impact on global disease burden. Under the background of climate change, rainstorms increase and floods occur frequently. Most studies show that the incidences of infectious diarrhea disease increase significantly after rainstorm and flood events. However, there is a lack of systematic summary on the path of rainstorm and flood events affecting the incidence of infectious diarrhea, including the key links and mechanisms underlying environmental-social interaction. This study comprehensively combed the literature from environmental factors, socio-economic and cultural factors, and population and individual susceptibility factors. The potential mechanisms of infectious diarrhea caused by rainstorm and flood events were discussed from the aspects of spreading of pathogens, affecting sanitation facilities and (or) drinking water treatment infrastructure, the regulatory role of individual and behavioral factors, and long-term effects. Based on the "pressure-state-response" model, a social driving process model of rainstorm and flood leading to incidence of infectious diarrhea was constructed. This model could provide reference for future quantitative modeling and other research directions. It is helpful to guide the public health departments to accurately identify factors affecting the incidence of infectious diarrhea after rainstorm and flood, so as to take targeted intervention measures.

Objective: To investigate the alterations of the cerebral resting-state spontaneous neural activity in colorectal cancer patients with depressive symptoms. Methods: Thirty-three colorectal cancer patients (patient group) with depression and 43 healthy subjects (control group) underwent the resting state functional magnetic resonance imaging (rs-fMRI). The amplitude of low-frequency fluctuations (ALFF) and fractional ALFF (fALFF) were calculated. Two independent samples t test were used to compare the ALFF and fALFF values between two groups by DPABI software, and then correlation analysis was performed between ALFF and fALFF with statistical significance and Patient Health Questionnaire (PHQ-9) scores and Generalized Anxiety Disorder (GAD-7) scores. Results: Compared with the control group, the patient group showed significantly lower ALFF and fALFF values in the bilateral precuneus, calcarine gyrus, lingual gyrus, left cuneus, superior, middle, inferior occipital gyrus and right fusiform gyrus (t=-5.730, P<0.05; t=-4.872, P<0.05). There were no significant correlations between the ALFF and fALFF values in these regions and PHQ-9 or GAD-7 scores (P>0.05). Conclusion Spontaneous decrease of neural activity in occipital and parietal lobes exists in colorectal cancer patients with depression at resting-sate, which may be a potential neurobiological marker.

Objective? To investigate the status of caregiver preparedness and adult attachment among the spouse of young and middle-aged stroke patients,and to analyze the correlation between them. Methods? From January 2017 to May 2018,110 spouses of stroke patients who were treated in Affiliated Hangzhou First People＇s Hospital, Zhejiang University School of Medicine were selected as subjects by convenience sampling. The General Information Questionnaire,Care Preparedness Scale (CPS),and Experience in Close Relationship Scale (ECR) were used in the survey. Totally 110 questionnaires were distributed, and 102 valid ones were retrieved, yielding an effective recovery rate of 92.73%. Results? The total CPS score of 102 young and middle-aged stroke patients was (12.87±5.39). The scores of attachment anxiety and attachment avoidance in ECR were (57.96 ±11.69) and (48.06 ±12.10). There were statistically significant differences in the scores of preparedness of caregivers among spouse caregivers of different ages, self reported health status and nursing experience (P＜ 0.05). Attachment anxiety dimension was negatively correlated with caregiver preparedness score (r=-0.318,P＜0.01), and attachment avoidance dimension was negatively correlated with caregiver preparedness score (r=-0.233,P＜ 0.01). Conclusions? The preparedness status of the spouses caregivers of young and middle-aged stroke patients are at a low level. Medical workers can improve their care preparedness by giving them home care knowledge guidance,reducing their attachment avoidance and attachment anxiety so as to improve their preparedness.

ObjectiveTo investigate the acceptance and influencing factors of manual hyperinflation (MIH) in patients with open sputum suction after tracheotomy, so as to provide reference for improving the acceptance of MIH for patients. MethodsBy convenience sampling, 140 patients with tracheotomy on open suction in Affiliated Hangzhou First People＇s Hospital, Zhejiang University School of Medicine from October 2016 to September 2018 were selected as participants in the study and the Self-Designed Questionnaire was used to study the acceptance towards MIH of the patients. Univariate analysis and Logistics regression analysis were conducted to analyze the influencing factors of the acceptance of patients on MIH. A total of 140 questionnaires were distributed, and 134 valid ones were collected. ResultsUnivariate analysis showed that there were statistical difference in age, gender, family support, duration of tracheotomy, whether the medical staff had explained the relevant knowledge of MIH, and whether the patients had experienced MIH on patients＇acceptance of MIH (P＜ 0.05). The above 6 statistically significant factors were analyzed by multivariate Logistic regression and the results showed that age, gender, duration of tracheotomy, whether the patients had experienced MIH before were the influencing factors of the patients＇ acceptance on MIH(P＜ 0.05). ConclusionsAge, gender, duration of tracheotomy, whether or not the patents have experienced MIH are independent impact factors for the patients＇ acceptance of MIH and the above factors can be the cut-in points of explanation and education in the process of MIH,so as to improve the patients＇ acceptance of MIH and promote recovery.