BACKGROUND:β-Caryophyllene has a variety of pharmacological activities such as antioxidant,anti-inflammatory and anti-apoptotic,which may have a better therapeutic effect on osteoarthritis.OBJECTIVE:To investigate the effect and mechanism of β-caryophyllene on mouse osteoarthritis.METHODS:Forty C57BL/6J mice were randomly divided into sham group,model group,low-dose β-caryophyllene group and high-dose β-caryophyllene group,with 10 mice in each group.Hulth method was used to construct an osteoarthritis model in the latter three groups.Four weeks after modeling,70 and 140 mg/kg/d β-caryophyllene was intragastrically given in the low-and high-dose β-caryophyllene groups,respectively,and normal saline was given by gavage in the sham group and the model group,once a day,for 4 weeks.After administration,knee joint morphological changes were observed by hematoxylin-eosin staining,serum levels of inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6,and interleukin-10)were detected by ELISA,and oxidative stress indexes(glutathione peroxidase,superoxide dismutase,and malondialdehyde)were detected by chemiluminescence.The expression levels of key proteins in the Sonic hedgehog(Shh)/glioma associated oncogene homolog 1(Gli1)signaling pathway were detected by immunohistochemistry and western blot.RESULTS AND CONCLUSION:(1)Compared with the sham group,a large number of inflammatory cells infiltrated in the knee joint of mice in the model group,cartilage tissue was seriously damaged,serum levels of tumor necrosis factor-α,interleukin-1β,interleukin-6,interleukin-10 and malondialdehyde were significantly increased(P＜0.01),the activities of glutathione peroxidase and superoxide dismutase were significantly decreased(P＜0.01),and the relative expression levels of Shh and Gli1 in the knee joint were significantly increased(P＜0.01).(2)Compared with the model group,in the low-and high-dose β-caryophyllene groups,inflammatory cell infiltration in the mouse knee joint was decreased,cartilage tissue injury was alleviated,serum levels of tumor necrosis factor-α,interleukin-1 β,interleukin-6 and malondialdehyde were significantly decreased(P＜0.05),the activities of glutathione peroxidase and superoxide dismutase were significantly increased(P＜0.01),and the expression levels of Shh and Gli1 in the knee joint were significantly decreased(P＜0.01).The above-mentioned improvements were more significant in the high-dose β-caryophyllene group than the low-dose β-caryophyllene group.To conclude,β-caryophyllene can improve osteoarthritis,and its mechanism may be related to reducing inflammation and oxidative stress damage by regulating the Shh/Gli1 signaling pathway.

BACKGROUND:H-type vessels(CD31hiEmcnhi)possess unique functionalities that offer new perspectives and entry points for comprehensively understanding the mechanisms of vascular-mediated bone metabolism regulation.This has triggered a significant paradigm shift in bone science research from a"bone-centric"approach to an"integrative bone-vascular"framework.OBJECTIVE:To perform a visual analysis based on literature data in the field of H-type vessels,aiming to identify research hotspots and emerging trends.METHODS:A systematic search was conducted in the CNKI and Web of Science databases for English and Chinese publications related to H-type vessels,covering the period from 2014 to 2024.The CiteSpace 6.2.R4 information visualization software was utilized to analyze and visualize data on publication countries,authors,institutions,keywords,and timeline views.RESULTS AND CONCLUSION:(1)A total of 59 Chinese and 185 English publications were included in the analysis.Since the introduction of the H-type vessel concept in 2014,the number of related research publications has been steadily increasing,accompanied by a significant growth in academic output.China leads the research in this domain,with major institutions including Southern Medical University,Sichuan University,and Shanghai Jiao Tong University.Keyword analysis indicated that current research hotspots primarily focus on the generation and regulatory mechanisms of H-type vessels,their roles in bone formation and remodeling,and their associations with bone metabolic diseases.Additionally,emerging keywords such as"induced membrane,""cartilage repair,"and"traditional Chinese medicine"suggest that research is progressively moving towards diversification and multidisciplinary integration.(2)H-type vessels play a crucial role in the development and progression of bone diseases,and relevant studies are pivotal for a deeper understanding of the physiological and pathological processes of bone tissue.Future research should further elucidate the specific mechanisms of H-type vessels in various disease contexts and promote the clinical translation of basic research findings.These efforts will provide innovative ideas and strategies for the prevention and treatment of bone-related diseases.

BACKGROUND:β-Caryophyllene has a variety of pharmacological activities such as antioxidant,anti-inflammatory and anti-apoptotic,which may have a better therapeutic effect on osteoarthritis.OBJECTIVE:To investigate the effect and mechanism of β-caryophyllene on mouse osteoarthritis.METHODS:Forty C57BL/6J mice were randomly divided into sham group,model group,low-dose β-caryophyllene group and high-dose β-caryophyllene group,with 10 mice in each group.Hulth method was used to construct an osteoarthritis model in the latter three groups.Four weeks after modeling,70 and 140 mg/kg/d β-caryophyllene was intragastrically given in the low-and high-dose β-caryophyllene groups,respectively,and normal saline was given by gavage in the sham group and the model group,once a day,for 4 weeks.After administration,knee joint morphological changes were observed by hematoxylin-eosin staining,serum levels of inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6,and interleukin-10)were detected by ELISA,and oxidative stress indexes(glutathione peroxidase,superoxide dismutase,and malondialdehyde)were detected by chemiluminescence.The expression levels of key proteins in the Sonic hedgehog(Shh)/glioma associated oncogene homolog 1(Gli1)signaling pathway were detected by immunohistochemistry and western blot.RESULTS AND CONCLUSION:(1)Compared with the sham group,a large number of inflammatory cells infiltrated in the knee joint of mice in the model group,cartilage tissue was seriously damaged,serum levels of tumor necrosis factor-α,interleukin-1β,interleukin-6,interleukin-10 and malondialdehyde were significantly increased(P＜0.01),the activities of glutathione peroxidase and superoxide dismutase were significantly decreased(P＜0.01),and the relative expression levels of Shh and Gli1 in the knee joint were significantly increased(P＜0.01).(2)Compared with the model group,in the low-and high-dose β-caryophyllene groups,inflammatory cell infiltration in the mouse knee joint was decreased,cartilage tissue injury was alleviated,serum levels of tumor necrosis factor-α,interleukin-1 β,interleukin-6 and malondialdehyde were significantly decreased(P＜0.05),the activities of glutathione peroxidase and superoxide dismutase were significantly increased(P＜0.01),and the expression levels of Shh and Gli1 in the knee joint were significantly decreased(P＜0.01).The above-mentioned improvements were more significant in the high-dose β-caryophyllene group than the low-dose β-caryophyllene group.To conclude,β-caryophyllene can improve osteoarthritis,and its mechanism may be related to reducing inflammation and oxidative stress damage by regulating the Shh/Gli1 signaling pathway.

BACKGROUND:H-type vessels(CD31hiEmcnhi)possess unique functionalities that offer new perspectives and entry points for comprehensively understanding the mechanisms of vascular-mediated bone metabolism regulation.This has triggered a significant paradigm shift in bone science research from a"bone-centric"approach to an"integrative bone-vascular"framework.OBJECTIVE:To perform a visual analysis based on literature data in the field of H-type vessels,aiming to identify research hotspots and emerging trends.METHODS:A systematic search was conducted in the CNKI and Web of Science databases for English and Chinese publications related to H-type vessels,covering the period from 2014 to 2024.The CiteSpace 6.2.R4 information visualization software was utilized to analyze and visualize data on publication countries,authors,institutions,keywords,and timeline views.RESULTS AND CONCLUSION:(1)A total of 59 Chinese and 185 English publications were included in the analysis.Since the introduction of the H-type vessel concept in 2014,the number of related research publications has been steadily increasing,accompanied by a significant growth in academic output.China leads the research in this domain,with major institutions including Southern Medical University,Sichuan University,and Shanghai Jiao Tong University.Keyword analysis indicated that current research hotspots primarily focus on the generation and regulatory mechanisms of H-type vessels,their roles in bone formation and remodeling,and their associations with bone metabolic diseases.Additionally,emerging keywords such as"induced membrane,""cartilage repair,"and"traditional Chinese medicine"suggest that research is progressively moving towards diversification and multidisciplinary integration.(2)H-type vessels play a crucial role in the development and progression of bone diseases,and relevant studies are pivotal for a deeper understanding of the physiological and pathological processes of bone tissue.Future research should further elucidate the specific mechanisms of H-type vessels in various disease contexts and promote the clinical translation of basic research findings.These efforts will provide innovative ideas and strategies for the prevention and treatment of bone-related diseases.

Organophosphorus pesticides(OPs)are widely used in global agriculture,and pose a serious threat to ecological environment and human health due to their high environmental persistence and biological toxicity.Colorimetric sensing strategies based on the inhibition of acetylcholinesterase(AChE)have become an important method for detecting OPs because of their simplicity and high specificity.However,the sensitivity is limited by the insufficient catalytic efficiency of traditional nanozymes.In this study,a one-step solvothermal method was used to synthesize polyoxometalate nanoribbons(POM)loaded with gold nanoparticles(Au NPs),named Au-POM.Experimental results showed that Au-POM could catalyze the decomposition of H2O2 in an acidic environment(pH 4.0),demonstrating typical peroxidase-like activity.Based on this,an AChE,choline oxidase(ChOx)and Au-POM nano enzyme cascade catalytic system was constructed.In this system,AChE specifically catalyzed the hydrolysis of acetylcholine(ACh)to choline,and then ChOx mediated the oxidation of choline to produce H2O2.During this process,Au-POM acted as a peroxidase-like enzyme to catalyze the decomposition of H2O2 to generate reactive oxygen species,triggering a specific oxidation reaction of the chromogenic substrate 3,3',5,5'-tetramethylbenzidine(TMB)into oxidized form.When the OP pesticide ethoprophos(EP)was present,it inhibited the activity of AChE and blocked the generation of ACh and H2O2,indirectly inhibiting the oxidation of TMB.The color and absorbance of the solution changed in a concentration-dependent manner.The detection limit of this method for EP was 1.05 μmol/L,and the linear response range was 20-180 μmol/L(R2=0.998).This method was applied to detection of environmental water samples and coriander samples with satisfactory results,providing a reliable technical platform for monitoring of OPs in environment and food.

Objective:To investigate whether neoadjuvant therapy can improve the prognosis of patients with pancreatic cancer.Methods:A retrospective case-control study analyzed data from the Surveillance, Epidemiology, and End Results (SEER) database on 12, 103 patients who underwent surgical treatment between January 1, 2010, and December 31, 2021. Patients were divided into the neoadjuvant therapy group ( n=3 276) and the upfront surgery group ( n=8 827) based on whether they received neoadjuvant treatment. The neoadjuvant therapy group included 2 342 patients receiving neoadjuvant chemotherapy and 934 patients receiving neoadjuvant chemoradiotherapy. The upfront surgery group consisted of 4 335 patients receiving adjuvant chemotherapy, 1 987 patients receiving adjuvant chemoradiotherapy, 63 patients receiving adjuvant radiotherapy, and 2 442 patients undergoing surgery alone. Propensity score matching was used to eliminate group differences and create a cohort with no statistical differences in other clinicopathological features except for the grouping variable. Variables such as age, gender, tumor location, race, population of residence, tumor diameter, household income, TNM stage, and information on radiotherapy and chemotherapy were used for 1∶1 case matching. T stage, N stage, and the use of radiotherapy or chemotherapy were matched exactly. After matching, 1 182 patients were included in each group: the neoadjuvant therapy group contained 1 155 patients receiving neoadjuvant chemoradiotherapy and 27 receiving neoadjuvant chemotherapy, while the upfront surgery group comprised 848 patients receiving adjuvant chemotherapy and 334 receiving adjuvant chemoradiotherapy. TNM staging was reported according to the 7th edition of the AJCC guidelines. The primary outcome was overall survival. Measurement data with skewed distributions were expressed as M( Q1, Q3), and intergroup comparisons were conducted using the Wilcoxon rank-sum test. Categorical data were compared using the chi-square test or the Fisher′s exact test. The Log-rank test and subgroup analyses to assess interactions between neoadjuvant therapy and subgroup in COX regression models were used to compare survival benefits across variables. Landmark analysis was performed to create segmented survival curves, studying the impact of neoadjuvant therapy on prognosis during different follow-up periods. Results:The neoadjuvant therapy group had a higher proportion of T 4 tumor involving celiac axis, superior mesenteric artery, and/or common hepatic artery compared to the upfront surgery group (14.7% vs 2.8%, P<0.001). Additionally, significant differences were observed between groups in terms of race, location, population of residence, age, tumor diameter, tumor stage, and adjuvant therapy regimen ( P<0.05). The median overall survival time in the neoadjuvant therapy group was 30 months, compared to 22 months in the upfront surgery group ( P<0.001). In the neoadjuvant therapy group, the median survival was 30 months for both neoadjuvant chemotherapy and chemoradiotherapy patients; in the upfront surgery group, it was 26 months for both adjuvant chemotherapy and chemoradiotherapy patients, 17 months for adjuvant radiotherapy patients, and 12 months for surgery-only patients. After propensity score matching, there were no differences in the distribution of clinical characteristics between groups ( P>0.05), and all patients in the matched cohort had received chemotherapy. The matched neoadjuvant therapy group had a longer median overall survival compared to the upfront surgery group (30 months vs 27 months, P<0.001). Subgroup interaction analysis revealed that T stage had a significant interaction with neoadjuvant therapy, both before (T 4 stage: HR=0.382, 95% CI: 0.319-0.458; T 2-T 3 stages: HR=0.696, 95% CI: 0.656-0.738; T 1 stage: HR=1.199, 95% CI: 0.867-1.657; interaction P<0.001) and after matching (T 4 stage: HR=0.581, 95% CI: 0.414-0.814; T 2-T 3 stages: HR=0.827, 95% CI: 0.734-0.931; T 1 stage: HR=1.320, 95% CI: 0.716-2.433; interaction P=0.043). Subgroup interaction analysis indicated that T 1 patients did not benefit from neoadjuvant therapy; survival curves plotted for matched T 1 patients showed no difference in survival between the neoadjuvant therapy group and the upfront surgery group ( P=0.323). Conversely, non-T 1 (T 2-T 4) stage patients showed significant survival benefits in both unmatched and matched cohorts ( P<0.001). Landmark analysis showing that the survival benefits occurred mainly in the early postoperative period of up to 3 years ( P<0.001), but there was no difference in overall survival between the neoadjuvant therapy group and the upfront surgery group of >3 years ( P>0.05). Patients with Arterial invasion (T 4 stage compared to T 1-T 3 stages) showed a similarly significant interaction with the benefit of neoadjuvant therapy in both the pre-matching cohort (interaction P<0.001) and the post-matching cohort (interaction P=0.037). Patients with T 4 stage disease in the neoadjuvant therapy group had longer overall survival compared to the upfront surgery group (median overall survival in pre-matching cohort: 30 months vs 13 months, P<0.001; median overall survival in post-matching cohort: 28 months vs 18 months, P=0.001). Among T 4 stage patients in the post-matching cohort, neoadjuvant therapy provided significant survival benefits during the early postoperative period of up to 3 years ( P=0.001). However, there was no difference in overall survival between the neoadjuvant therapy group and the direct surgery group beyond 3 years( P=0.729). Conclusions:The prognosis in the neoadjuvant therapy group was better than in the upfront surgery group. Propensity score matching and subgroup interaction analysis showed that non-T 1 and T 4 stage patients benefited more from neoadjuvant therapy, with benefits mainly seen in the early postoperative period (≤3 years).

Objective:To investigate the liver-protective effect of Peipi Shugan Decoction (PSD) in rats with liver fibrosis through network pharmacology and experimental animal models.Methods:TCMSP was used to retrieve the active components of Peipi Shugan Decoction, GeneCards database was used to obtain the liver fibrosis related targets, and Venny 2.1 was used to obtain the intersection targets. A protein-protein interaction (PPI) network was constructed using the STRING 12.0 database, and Go function and KEGG pathway enrichment analysis were performed through David database. A total of 60 male Sprague-Dawley (SD) rats were divided into two groups: a blank control group ( n=10) and a model establishment group ( n=50) with random number table method. Liver fibrosis models were induced in the model group by intraperitoneal injection of a 40% CCl?- olive oil solution. After successful modeling, the rats were randomly assigned to the following groups ( n=10 per group): model group, colchicine group, and Peipi Shugan Decoction low-, medium-, and high-dosage groups. The colchicine group received colchicine suspension at 0.2 ml/kg via intragastric administration. Peipi Shugan Decoction low-, medium-, and high-dosage groups were administered the decoction at dosages of 2.81, 5.63, and 11.25 g/kg, respectively. The blank control and model groups received an equal volume of normal saline. All treatments were administered once daily for 8 consecutive weeks. HE staining and Masson staining were used to observe the morphological changes of liver tissue and the deposition of collagen fibers. he levels of GPT, GOT and ALP were detected by automatic biochemical analyzer. The expressions of JAK2/STAT1 signaling pathway related proteins, α - smooth muscle actin (α-SMA) and Collagen Ⅰ in rat liver were detected by Western blot. The expression of α-SMA protein in liver tissue was detected by immunohistochemistry. The number and ratio of CD4 + T cells and CD8 + T cells in liver tissue were analyzed by flow cytometry. The levels of IL-6, IL-1β and TNF-α in serum were detected by ELISA. Results:A total of 94 active components were screened out from Peipi Shugan Decoction; the prediction analysis results revealed that this compound formula shared 122 common targets with liver fibrosis diseases; The top five core targets with degree values in the PPI network are AKT1, TNF, IL-6, TP53, and IL-1β. GO enrichment analysis further indicated that Peipi Shugan Decoction mainly achieved anti-liver fibrosis effects by regulating JAK2/STAT1 signaling pathway and other mechanisms. Compared with the model group, the colchicine group and Peipi Shugan Decoction low-, medium-, and high-dosage groups exhibited decreased serum levels of GPT, GOT, ALP, IL-6, TNF-α, and IL-1β ( P<0.05), as well as reduced expression of α-SMA in liver tissue ( P<0.05). Peipi Shugan Decoction medium- and high-dosage groups showed increased protein expression of p-STAT1/STAT1 in liver tissue ( P<0.05), while Peipi Shugan Decoction high-dosage group demonstrated decreased α-SMA protein expression ( P<0.05). Additionally, Peipi Shugan Decoction medium- and high-dosage groups exhibited reduced expressions of CD4 + and CD8 + ( P<0.05). Conclusions:Peipi Shugan Decoction has the characteristics of multi-component, multi-target and multi-pathway in the treatment of liver fibrosis. Its mechanism is mainly related to activating the JAK2/STAT1 signaling pathway, inhibiting cellular inflammatory responses and hindering the activation of hepatic stellate cells (HSC).

Objective To investigate the effects of electroacupuncture at Baihui(GV20)point and Dazhui(GV14)point on cognitive function and potential mechanisms in Alzheimer's disease(AD)mice.Methods Forty-eight male five-transgenic familial Alzheimer's disease(5xFAD)mice were divided into four groups:empty vector group,knockdown group,empty vector+electroacupuncture group,and knockdown+electroacupuncture group.The knockdown group received Rho-associated coiled-coil containing protein kinase 2(ROCK2)knockdown via adeno-associated virus(AAV)microinjection technology.Knockdown efficiency was verified by polymerase chain reaction(PCR),and gene expression levels were confirmed by Western Blot.After 4 weeks of viral expression and transfection,electroacupuncture intervention at Baihui and Dazhui acupoints was administered for 4 weeks.Cognitive function was assessed using behavioral tests.Pathological products,inflammatory markers[interleukin 6(IL-6),tumor necrosis factor α(TNF-α)],and changes in the PI3K/AKT signaling pathway were detected using immunofluorescence staining,enzyme-linked immunosorbent assay(ELISA),and Western Blot techniques.Results Compared with the empty vector group,the ROCK2 knockdown group showed significantly improved spatial and contextual memory(P＜0.05 or P＜0.01),reduced amyloid deposition and inflammatory indicators(P＜0.05 or P＜0.01),and increased expression levels of phosphorylated protein kinase B(P-AKT)and phosphorylated phosphatidylinositol-3-kinase(P-PI3K)(P＜0.01),while the knockdown+electroacupuncture group exhibited more pronounced effects than knockdown group(P＜0.05 orP＜0.01).Conclusion Electroacupuncture at Baihui point and Dazhui point can regulate ROCK2 to activate the PI3K/AKT signaling pathway,improve cognitive impairment in AD mice,reduce Aβ deposition,and alleviate inflammatory responses.

BackgroundAlcohol dependence patients are prone to relapse after their attempts to quit drinking， which poses a considerable threat to their physical and mental health and creates a heavy burden on their families. Currently， empowerment education is increasingly being utilized in the rehabilitation management of chronic diseases， but there remains a striking lack of empirical research on the application of "Internet +" empowerment education based on timing it right in alcohol dependence patients. ObjectiveTo explore the impact of "Internet +" empowerment education based on timing it right on patients with alcohol dependence， in order to maximize the reduction in relapse rates， craving for alcohol and severity of anxiety symptoms. MethodsA total of 120 patients who were hospitalized in the Department of Addiction Medicine， Hebei Provincial Mental Health Center from May 2022 to April 2023 and met the diagnostic criteria for alcohol dependence in the International Classification of Diseases， tenth edition （ICD-10） were enrolled， and they were classified into study group （n=62） and control group （n=58） using random number table methods. Both groups received standard medication and routine care. Additionally， study group underwent a 6-month "Internet +" empowerment education based on timing it right. At baseline， all subjects were assessed using Penn Alcohol Craving Scale （PACS） and Self-rating Anxiety Scale （SAS）. Three months and six months after intervention， assessments were conducted using PACS， SAS and Michigan Alcoholism Screening Test （MAST）. ResultsThe relapse rates after three and six months of intervention were both lower in study group than those in control group， with statistically significant differences （χ²=8.575， 8.828， P<0.01）. ANOVA with repeated measures on PACS total score and scores of each item revealed a significant time effect， group effect and time×group interaction effect （F=159.714~837.751， 84.645~393.606， 24.302~137.896， P<0.01）. And significant time effect， group effect and time×group interaction effect were also reported on SAS scores （F=166.237， 65.325， 24.724， P<0.01）. Conclusion"Internet +" empowerment education based on timing it right may help reduce relapse rates， alcohol cravings and severity of anxiety symptoms among patients with alcohol dependence. ［Funded by 2023 Annual Hebei Provincial Medical Scientific Research Project Plan （number， 20231537）］

Diterpenoid ferruginol is a key intermediate in biosynthesis of active ingredients such as tanshinone and carnosic acid.However, the traditional process of obtaining ferruginol from plants is often cumbersome and inefficient. In recent years, the increasingly developing gene editing technology has been gradually applied to the heterologous production of natural products, but the production of ferruginol in microbe is still very low, which has become an obstacle to the efficient biosynthesis of downstream chemicals, such as tanshinone. In this study, miltiradiene was produced by integrating the shortened diterpene synthase fusion protein,and the key genes in the MVA pathway were overexpressed to improve the yield of miltiradiene. Under the shake flask fermentation condition, the yield of miltiradiene reached about(113. 12±17. 4)mg·L~(-1). Subsequently, this study integrated the ferruginol synthase Sm CYP76AH1 and Sm CPR1 to reconstruct the ferruginol pathway and thereby realized the heterologous synthesis of ferruginol in Saccharomyces cerevisiae. The study selected the best ferruginol synthase(Il CYP76AH46) from different plants and optimized the expression of pathway genes through redox partner engineering to increase the yield of ferruginol. By increasing the copy number of diterpene synthase, CYP450, and CPR, the yield of ferruginol reached(370. 39± 21. 65) mg·L~(-1) in the shake flask, which was increased by 21. 57-fold compared with that when the initial ferruginol strain JMLT05 was used. Finally, 1 083. 51 mg·L~(-1) ferruginol was obtained by fed-batch fermentation, which is the highest yield of ferruginol from biosynthesis so far. This study provides not only research ideas for other metabolic engineering but also a platform for the construction of cell factories for downstream products.
Saccharomyces cerevisiae/genetics* ; Diterpenes/metabolism* ; Metabolic Engineering ; Fermentation ; Abietanes