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 establish UPLC fingerprint method and 2 contents determination methods of Buddleja officinalis; To provide a reference for improving the quality control standard and evaluation of Buddleja officinalis from different habitats.Methods:UPLC method was used to establish the fingerprints of 17 batches of Buddleja officinalis. The similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to compare the quality differences of Buddleja officinalis from different habitats. The contents of acteoside and linarin in Buddleja officinalis were determined.Results:There were 12 common peaks in UPLC fingerprints of Buddleja officinalis, six of which were identified as echinacoside, acteoside, cynaroside, isoacteoside, linarin, and apigenin. The fingerprint similarity of 17 batches of Buddleja officinalis was more than 0.9; Buddleja officinalis from different habitats were classified into 2 groups. Five differential markers were determined by OPLS-DA analysis. The order of significance was acteoside > peak 3 > echinacoside > isoacteoside > linarin. Edgeworthia chrysantha was identified by the method of fingerprint as counterfeit. The results of content determination showed that the content of Buddleja officinalis in Hubei and Sichuan was the high and stable.Conclusion:The method can effectively analyze the differences of Buddleja officinalis from different habitats, and provide reference for the quality control of Buddleja officinalis.

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.

Objective:To establish the ultra high performance liquid chromatography (UPLC) fingerprints of Mume flos at different flowering stages; To provide reference for the quality research of Mume flos.Methods:The fingerprints of Mume flos were established by UPLC method, and the common peaks were identified by high performance liquid chromatography high resolution mass spectrometry (LC-MS). Chemometrics analysis was carried out with the fingerprints' common peak area of plum blossom at different flowering stages as a variable. Semiquantitative analysis of changes in flavonoids and phenolic acids in Mume flos at different flowering stages was conduct using peak area calculation method.Results:Totally 31 common peaks were identified in the fingerprints of plum blossom medicinal materials at different flowering stages and 9 components were identified. Clustering analysis (HCA) and principal component analysis (PCA) both classified plum blossom medicinal herbs at different flowering stages into three categories. Among them, there were significant differences between the groups at the bud stage, blooming period, and final flowering period, while the differences between the groups at blooming period and final flowering period were relatively small. The orthogonal partial least squares discriminant analysis (OPLS-DA) screened 16 different components with VIP>1.0. The contents of phenolic acids in different flowering stages were as follows: bud stage>blooming period>final flowering period, while the contents of flavonoids were as follows: blooming period>final flowering period>bud stage.Conclusions:This method is simple and reliable, and can provide reference for the quality evaluation of plum blossom medicinal materials at different flowering stages.

Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

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 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 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.