BACKGROUND:Previous studies have found that neohesperidin can delay bone loss in ovariectomized mice and has the potential to treat osteoporosis,but its specific mechanism of action remains to be explored. OBJECTIVE:To explore the key targets and possible mechanisms of neohesperidin in the treatment of osteoporosis based on bioinformatics and cell experiments in vitro. METHODS:The gene expression dataset related to osteoporosis was obtained from GEO database,and the differentially expressed genes were screened and analyzed in R language.The osteoporosis-related targets were screened from GeneCards and DisGeNET databases,and the neohesperidin-related targets were screened from ChEMBL and PubChem databases,and the common targets were obtained by intersection of the three.The String database was used to construct the PPI network of intersection genes,and the key targets were screened.The DAVID database was used for GO and KEGG enrichment analysis.The AutoDock software was used to verify the molecular docking between the neohesperidin and the target protein.The effect of neohesperidin on osteogenic differentiation of C57 mouse bone marrow mesenchymal stem cells was detected.Complete medium was used as blank control group;osteogenic induction medium was used as the control group;and osteogenic induction medium containing different concentrations of neohesperidin(25,50 μmol/L)was used as experimental group.The expression of alkaline phosphatase,the degree of mineralization,the expression of osteogenic-related genes and target genes during osteogenic differentiation of cells were measured at corresponding time points. RESULTS AND CONCLUSION:(1)9 253 differentially expressed genes,2 161 osteoporosis-related targets,and 326 neohesperidin-related targets were screened.There were 53 common targets among the three.All 53 genes were up-regulated in osteoporosis samples.The PPI network screened the target gene PRKACA of research significance.GO function and KEGG pathway enrichment analysis showed that neohesperidin's treatment of osteoporosis through PRKACA target mainly depended on biological processes such as protein phosphorylation and protein autophosphorylation,acting on endocrine resistance,proteoglycan in cancer,and estrogen signaling pathway to play a therapeutic role.Molecular docking results showed that neohesperidin had a certain binding ability to the protein corresponding to the target PRKACA.(2)The results of alkaline phosphatase staining showed that neohesperidin could promote the expression of alkaline phosphatase in the early stage of osteogenic differentiation of mesenchymal stem cells.Alizarin red staining showed that neohesperidin could promote the mineralization of osteogenic differentiation of mesenchymal stem cells.RT-qPCR results showed that neohesperidin could increase the mRNA expression of alkaline phosphatase,PRKACA,and osteocalcin.(3)These results indicate that neohesperidin may promote osteogenic differentiation through PRKACA target on the estrogen signaling pathway to prevent and treat osteoporosis.

Objective:To observe the long-term effect of superficial temporal fascia flap combined with severed auricle reimplantation in emergency repair of partial auricle of complete separation.Methods:The data of patients with partial auricle of complete separation admitted to Emergency Clinic of Burn and Plastic Surgery of General Hospital of Northern Theater Command from June 2014 to August 2023 were retrospectively analyzed. All of them were repaired with a superficial temporal fascia flap combined with amputating auricle reimplantation. During the operation, the superficial temporal fascia flap was harvested, and the pedicle was preserved. Then the detached auricular cartilage was removed and used as a replantation scaffold. Then the remaining skin was thinned to create a full-thickness skin graft after cartilage detachment. Referring to the position and angle of the contralateral auricle, the cartilage scaffold was sutured and fixed at the stump of the ear cartilage. The wound was covered with a superficial temporal fascia flap and a full-thickness skin graft, and then packed and sutured. The postoperative observation indicators mainly confirm whether the surgery was successful, the healing condition of the replanted ear, and whether there were problems such as skin flap necrosis, infection, hematoma, etc. The shape, color, texture and tactile recovery of the reconstructed auricle were evaluated by long-term follow-up for more than 1 year. The Vancouver scar scale (VSS) was used to assess scarring in both donor and recipient sites (total score of 0-15 points, higher scores indicated more severe scarring). The Likert 5-level scoring method was used to evaluate the patients’ satisfaction with the surgical results (total score of 30 points, ≥27 points were very satisfied, 24-26 points were somewhat satisfied, 18-23 points were indifferent, 15-17 points were somewhat dissatisfied, ≤14 points were very dissatisfied).Results:A total of 8 patients were enrolled, including 5 males and 3 females. Their ages ranged from 26 to 65 years, with an average of 41 years. All patients had unilateral ear defects, with 3 cases in the left ear and 5 cases in the right ear. The defect areas ranged from 1.5 cm × 2.5 cm to 5.0 cm × 4.0 cm. During the surgery, the harvested superficial temporoparietal fascia flaps ranged from 4.5 cm × 6.5 cm to 15.0 cm × 10.0 cm. After surgery, both the flaps and full-thickness skin graft healed satisfactorily, with primary healing observed in both the donor and recipient sites. There were no complications such as necrosis, infection, or hematoma were observed. The follow-up period ranged from 3 to 9 years, with an average of approximately 6.3 years. Except for one case that required a defatted surgery at the second stage, the reconstructed auricles of the remaining patients were basically consistent with the healthy side, with smooth contour lines, skin color and texture close to the surrounding tissues, and improved tactile sensitivity. In the final follow-up, the VSS scores for both the donor and recipient sites were ≤3 for all patients. All patients rated the surgical outcome as very satisfied.Conclusion:For the partial auricle of complete separation that has no chance of replantation, the use of superficial temporal fascia flap and detached ear composite graft to repair is a reliable and effective surgical method. Patients have good postoperative long-term effects and high levels of satisfaction.

Objective:To observe the long-term effect of superficial temporal fascia flap combined with severed auricle reimplantation in emergency repair of partial auricle of complete separation.Methods:The data of patients with partial auricle of complete separation admitted to Emergency Clinic of Burn and Plastic Surgery of General Hospital of Northern Theater Command from June 2014 to August 2023 were retrospectively analyzed. All of them were repaired with a superficial temporal fascia flap combined with amputating auricle reimplantation. During the operation, the superficial temporal fascia flap was harvested, and the pedicle was preserved. Then the detached auricular cartilage was removed and used as a replantation scaffold. Then the remaining skin was thinned to create a full-thickness skin graft after cartilage detachment. Referring to the position and angle of the contralateral auricle, the cartilage scaffold was sutured and fixed at the stump of the ear cartilage. The wound was covered with a superficial temporal fascia flap and a full-thickness skin graft, and then packed and sutured. The postoperative observation indicators mainly confirm whether the surgery was successful, the healing condition of the replanted ear, and whether there were problems such as skin flap necrosis, infection, hematoma, etc. The shape, color, texture and tactile recovery of the reconstructed auricle were evaluated by long-term follow-up for more than 1 year. The Vancouver scar scale (VSS) was used to assess scarring in both donor and recipient sites (total score of 0-15 points, higher scores indicated more severe scarring). The Likert 5-level scoring method was used to evaluate the patients’ satisfaction with the surgical results (total score of 30 points, ≥27 points were very satisfied, 24-26 points were somewhat satisfied, 18-23 points were indifferent, 15-17 points were somewhat dissatisfied, ≤14 points were very dissatisfied).Results:A total of 8 patients were enrolled, including 5 males and 3 females. Their ages ranged from 26 to 65 years, with an average of 41 years. All patients had unilateral ear defects, with 3 cases in the left ear and 5 cases in the right ear. The defect areas ranged from 1.5 cm × 2.5 cm to 5.0 cm × 4.0 cm. During the surgery, the harvested superficial temporoparietal fascia flaps ranged from 4.5 cm × 6.5 cm to 15.0 cm × 10.0 cm. After surgery, both the flaps and full-thickness skin graft healed satisfactorily, with primary healing observed in both the donor and recipient sites. There were no complications such as necrosis, infection, or hematoma were observed. The follow-up period ranged from 3 to 9 years, with an average of approximately 6.3 years. Except for one case that required a defatted surgery at the second stage, the reconstructed auricles of the remaining patients were basically consistent with the healthy side, with smooth contour lines, skin color and texture close to the surrounding tissues, and improved tactile sensitivity. In the final follow-up, the VSS scores for both the donor and recipient sites were ≤3 for all patients. All patients rated the surgical outcome as very satisfied.Conclusion:For the partial auricle of complete separation that has no chance of replantation, the use of superficial temporal fascia flap and detached ear composite graft to repair is a reliable and effective surgical method. Patients have good postoperative long-term effects and high levels of satisfaction.

Objective To establish the ultra-high performance liquid chromatography(UPLC)chromatographic fingerprint of Notopterygii Rhizoma et Radix;To determine the contents of ferulic acid,nodakenin,ammijin,notopterol,isoimperatorin and volatile oil of Notopterygii Rhizoma et Radix from different producing areas;To provide reference for quality evaluation of Notopterygii Rhizoma et Radix.Methods Waters BEH C18 chromatographic column(2.1 mm×150 mm,1.7 μm)was used,with mobile phase acetonitrile-0.02%formic acid aqueous solution gradient elution,flow rate 0.25 mL/min,column temperature 25℃,detection wavelength 330 nm,injection volume 2 μL.UPLC fingerprints of 25 batches of Notopterygii Rhizoma et Radix were established,and the similarity analysis and chemometrics analysis were carried out.The contents of ferulic acid,nodakenin,ammijin,notopterol and isoimperatorin were determined simultaneously,and the contents of volatile oil was determined by steam distillation method.Results Totally 23 common fingerprint peaks were calibrated,11 known components were identified.According to the results of the cluster analysis and principal component analysis,25 batches of Notopterygii Rhizoma et Radix samples were divided into 3 categories,and the 6 potential differential components were screened out by orthogonal partial least squares-discriminant analysis(OPLS-DA).The results showed that the contents of notopterol and volatile oil from Sichuan Province were higher than those from Gansu Province and Qinghai Province.Conclusion The method established in the study is accurate and reliable,which can provide scientific basis and reference for the quality evaluation and control of Notopterygii Rhizoma et Radix.

Objective:To investigate the utility of 68Ga-fibroblast activation protein (FAP) inhibitor (FAPI)-04 PET imaging in assessing the therapeutic response in pressure overload-induced heart failure. Methods:Rat models of pressure overload-induced heart failure were established by abdominal aortic constriction (AAC). Thirty rats were categorized into AAC group, 7 days reverse AAC (rAAC) group, and sham operation (sham) group ( n=10 per group) using completely random grouping method. All rats underwent 68Ga-FAPI-04 PET/CT imaging at 4 and 8 weeks after the ACC operation, while echocardiography, pathological examination, and immunohistochemical analysis were performed at 8 weeks postoperation. One-way analysis of variance, independent-sample t test and Pearson correlation analysis were used for data analysis. Results:68Ga-FAPI-04 PET/CT imaging showed that the target-to-background ratios of the heart and liver had significant differences among three groups both at 4 and 8 weeks postoperation ( F values: 2 547.12, 2 041.71, 462.65, 1 210.97, all P<0.001). Echocardiography revealed left ventricular ejection fraction (LVEF), left ventricular internal diameter at end-diastole (LVIDd), and left ventricular internal diameter at end-systole (LVIDs) in three groups at 8 weeks postoperation were significantly different ( F values: 118.92, 9.11, 10.63, all P<0.01). Rats were sacrificed at 8 weeks postoperation, and Masson staining showed that the fibrosis in the heart and liver of the rAAC group was significantly improved compared with that of the AAC group, and immunohistochemical analysis revealed significantly lower FAP levels in the heart and liver of the rAAC group compared with those of the AAC group ( t values: from -11.27 to -4.16, all P<0.01). FAPI uptake in the heart of the AAC group and rAAC group at 8 weeks postoperation were significantly positively correlated with FAPI uptake in the liver, LVIDd and LVIDs, FAPI uptake in the heart was significantly negatively correlated with LVEF, and FAPI uptake in the heart and liver were significantly positively correlated with fibrosis degree and FAP levels of corresponding organs ( r values: -0.89, -0.88, 0.72-0.97, all P<0.05). Conclusions:68Ga-FAPI-04 PET/CT can show the improvement process of cardio-liver fibrosis following the unloading of excessive pressure in heart failure. Myocardial FAPI uptake is closely related to the extent of heart failure improvement.

Objective:To explore the feasibility of actively screening patients with chronic obstructive pulmonary disease (COPD) among inpatients in county territory-level hospitals based on the collection of comorbidity-related diseases and questionnaire surveys.Methods:This study was a cross-sectional study. From April 1, 2023, to November 30, 2023, a total of 1 392 inpatients who met the screening criteria in county territory-level hospitals within the Western Medical Group of Baiyun District, Guangzhou, were included in the study. General information, disease data, and COPD screening data of the patients were collected. A total of 1 392 questionnaires were distributed, all of which were returned and included in the analysis. Descriptive analysis, comparative analysis, and association rule mining were conducted, including the distribution of general information, distribution of common comorbidity-related diseases in COPD, distribution of questionnaire screening and pulmonary function test results, comparison of screening results based on comorbidity-related diseases grouping, comparison of screening results based on questionnaire screening results grouping, comparison of screening results based on smoking total score grouping, and association rules between screening results and pulmonary function test results and other research data.Results:Among the 1 392 study subjects, 334 cases (24.0%) had a positive self-screening questionnaire for COPD, 44 cases (13.2%) completed pulmonary function tests, and 17 cases (38.6%) were diagnosed with COPD. The positive rate of the screening questionnaire among inpatients was lowest in surgical patients without comorbidity-related diseases and highest in male patients with single/multiple comorbidity-related diseases and symptoms of chronic respiratory system diseases. The group with multiple comorbidity-related diseases had a significantly higher positive rate in the screening questionnaire than the group with single comorbidity-related diseases and the group without comorbidity-related diseases. Only 13.2% of inpatients with a positive screening questionnaire completed pulmonary function tests, with residents covered by medical insurance with multiple comorbidity-related diseases, including cardiovascular diseases, having the lowest rate, and patients with symptoms of chronic respiratory system diseases and single comorbidity-related diseases having the highest rate.Conclusions:Based on the collection of comorbidity-related diseases and questionnaire surveys, it is feasible to actively screen COPD patients among inpatients in county territory-level hospitals. However, efforts are needed to further increase the proportion of inpatients with positive screening questionnaires undergoing pulmonary function tests.

Objective:To analyze and compare the health education efforts of respiratory infectious diseases at home and abroad.Methods:The literature related to health education and popular science of respiratory infectious diseases included in the databases of China National Knowledge Infrastructure (CNKI) and Web of Science (WOS) from January 1, 2003 to December 31, 2023 was searched. A total of 4 686 articles were retrieved in CNKI, 1 540 articles unrelated to the theme were excluded, and 3 146 Chinese articles were obtained. In the WOS database, 7 724 articles were retrieved, 3 685 articles about the clinical mechanism of diseases, pharmacology, and other research topics were excluded, and 4 039 English articles were obtained. The information of annual publications, institutions, authors and keywords was analyzed by using CiteSpace visualization software, and the publication status, research hotspots and development trends of health education related to respiratory infectious diseases at home and abroad were analyzed.Results:Since 2003, the number of publications in health education on respiratory infectious diseases at home and abroad had shown a fluctuating growth trend, and in 2020, the field showed a sharp growth trend at home and abroad. There was no core author group in this field in China, and the network density of domestic authors was 0.006 5, and the network density of foreign authors was 0.009 6. The domestic institutions were mainly the Center for Disease Control and Prevention and medical institutions, including Guangzhou Chest Hospital (29 articles) and the Chinese Center for Disease Control and Prevention (10 articles); Foreign research institutions were mainly higher education institutions, including the University of London (91 articles) and Harvard University (67 articles). The network density of domestic was 0.001 3, and the network density of foreign publishing institutions was 0.026 3, the network density was greater than that of Chinese publishing institutions. The emergence of "COVID-19" "Avian influenza" "Knowledge, Attitude, Practice" and "mental health" in China had strong burst (burst intensity: 46.41, 12.12, 10.33, 8.5); "Coronavirus" "coverage" "Avian influenza" and "COVID-19 vaccine" in foreign countries had strong burst (burst intensity: 14.34, 11.06, 10.73, 10.02).Conclusions:At present, the health education of respiratory infectious diseases at home and abroad has received great attention. But the cooperation between domestic authors and research institutions is loose, and the close collaboration needs to be strengthened. There are differences in the research focus of health education on respiratory infectious diseases at home and abroad.

The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26％,and the intra-and inter-batch co-efficients of variation were both less than 10％.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.

Objective:To systematically evaluate the efficacy and safety of different antifungal medications for fungal keratitis (FK).Methods:A network meta-analysis was conducted.Four databases including PubMed, Cochrane Library, Embase, Web of Science were searched.The publication period was from inception to March 16, 2023.Two researchers followed the inclusion and exclusion criteria to screen randomized controlled trial (RCT), completed the quality assessment and extracted the information.Literature quality assessment was performed using Review Manager 5.4 bias risk assessment tool, and network meta-analysis was performed using Stata 14.0 software for cure rate, healing time, visual acuity improvement and safety of different antifungal drugs for fungal keratitis.Results:A total of 14 RCTs involving 1 681 patients were finally included in this study.The network meta-analysis showed that 0.2% chlorhexidine eye drops and 5% natamycin eye drops+ oral voriconazole had better efficacy than other interventions in cure rate, and the surface under the cumulative ranking (SUCRA) were 86.1% and 63.3%, respectively.The cure rate of 1% voriconazole eye drops was lower than that of 0.2% chlorhexidine eye drops, 5% natamycin eye drops+ oral voriconazole, 0.05% chlorhexidine eye drops, 0.1% chlorhexidine eye drops, 5% natamycin eye drops+ oral ketoconazole and 5% natamycin eye drops, showing statistically significant differences (all at P<0.05).1% voriconazole eye drops and 5% natamycin eye drops+ oral voriconazole showed better efficacy than other interventions in terms of healing time (SUCRA=66.9%, 55.7%).The combination of 5% natamycin eye drops, 1% voriconazole eye drops and oral voriconazole showed a better efficacy than others in improving visual acuity and safety (SUCRA=74.8%, 79.7%).For safety, 5% natamycin eye drops was superior to 1% voriconazole eye drops and 0.2% chlorhexidine eye drops, with statistically significant differences (both at P<0.05).In addition, there may be potential publication bias in this analysis. Conclusions:0.2% chlorhexidine eye drops and 1% voriconazole eye drops are effective in the treatment of FK.The combination use of 5% natamycin eye drops, 1% voriconazole eye drops and oral voriconazole can improve visual acuity and had good safety.

Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.