OBJECTIVE To establish characteristic chromatogram of Yao medicine Kadsura longipedunculata and the method for the content determination of its main component anwulignan， and evaluate the anti-inflammatory activity of anwulignan. METHODS HPLC method was performed with acetonitrile-0.5% phosphoric acid solution as the mobile phase for gradient elution. The characteristic chromatogram of K. longipedunculata was established and similarity was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. The content of anwulignan in K. longipedunculata was determined. Lipopolysaccharide induced RAW264.7 macrophages were selected as inflammatory cell model to investigate the effects of anwulignan on the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6. RESULTS The similarities of characteristic chromatogram for 10 batches of K. longipedunculata ranged 0.901-0.994， and 9 common peaks were determined； 3 components were identified， such as changnan schisantherin E， kadsulactone A， anwulignan. The contents of anwulignan were （0.72±0.05）-（1.21±0.03） mg/g（n＝3）. Anwulignan of 0.125-0.5 μg/mL greatly decreased the levels of TNF-α， IL-1β and IL-6 in the supernatant of inflammatory model cells （P＜0.05 or P＜0.01）. CONCLUSIONS HPLC characteristic chromatogram of K. longipedunculata and the method for the content determination of anwulignan are all established， and anwulignan may be the active ingredient of anti-inflammatory effect in K. longipedunculata.

Objective To explore the effect of etoricoxib combined with Bitong decoction formula on the clinical efficacy and serum level of patients with knee osteoarthritis.Methods Retrospective analysis was used to collect patients with knee osteoarthritis admitted to our hospital from August 2022 to August 2023.According to the medication method,they were divided into the combined group and the control group.The control group was given etoricoxib tablets,and the combination group was given a combination of Bitong decoction formula on the basis of the control group.Both groups were treated for one month.The efficacy after treatment,Traditional Chinese Medicine(TCM)syndrome score,the knee joint function[osteoarthritis index(WOMAC),Lequesne score],inflammatory indicators[interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-3(MMP-3),tissue inhibitor of metalloproteinase-1(TIMP-1)],oxidative stress indicators[total antioxidant capacity(TAOC),lipid peroxidation(LPO),nitric oxide(NO)]before and after treatment and safety of the two groups were compared.Results A total of 162 patients were included,with 81 in each group.The overall efficacy and total effective rate of the combined group were higher than those of the control group(P＜0.05).After treatment,TCM syndrome score,WOMAC,Lequesne score,IL-1 β,TNF-α,MMP-3,TIMP-1,LPO and NO levels in two groups were decreased compared with before treatment,while TAOC increased compared with before treatment(P＜0.05),and all the above indexes in the combination group were better than those in the control group(P＜0.05).The safety of the two groups was comparable(P＞0.05).Conclusion The efficacy of etoricoxib combined with Bitong decoction formula in knee osteoarthritis is better than that of single western medicine,which can alleviate symptoms and improve knee joint function,as well as reduce inflammation and oxidative stress,and has high safety.

Objective To isolate the secondary metabolites of endophytic fungus Aspergillus sp. J218 from Anectochilus roxburhii. Methods Different chromatographic methods, including Sephadex LH-20 and silica gel chromatography as well as HPLC, were used to isolate compounds from the EtOAc fraction of the solid fermentation of J218, and their structures were identified by spectral methods. Results Ten compounds were isolated from the fermentation of J218 and their structures were individually identified as kotanin（1）, flavasperone（2）, aurasperone B（3）, fonsecinone B（4）, fonsecinone D（5）, ensidol A（6）, fonsecinone A（7）, fonsecinone C（8）, aurasperone A（9）, and fonsecinone F（10）. Conclusion Most compounds isolated from endophytic fungus Aspergillus sp. J218 in Anectochilus roxburhii were identified as dimeric naphthopyrones. These results suggest that this strain contains rich dimerization synthase, which could provide clues for the further exploration of the rational biosynthesis pathway of dimeric naphthopyrones in this strain.

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Lower extremity deep vein thrombosis (DVT) is one of the main complications in patients with traumatic fractures, and for severe patients, the DVT can even affect arterial blood supply, resulting in insufficient limb blood supply. If the thrombus breaks off, pulmonary embolism may occur, with a high mortality. The treatment and rehabilitation strategies of thrombosis in patients with lower extremity fractures have its particularity. DVT in traumatic fractures patients has attracted extensive attention and been largely studied, and the measures for prevention and treatment of DVT are constantly developing. In recent years, a series of thrombosis prevention and treatment guidelines have been updated at home and abroad, but there are still many doubts about the prevention and treatment of DVT in patients with different traumatic fractures. Accordingly, on the basis of summarizing the latest evidence-based medical evidence at home and abroad and the clinical experience of the majority of experts, the authors summarize the clinical treatment and prevention protocols for DVT in patients with traumatic fractures, and make this consensus on the examination and assessment, treatment, prevention and preventive measures for DVT in patients with different fractures so as to provide a practicable approach suitable for China ′s national conditions and improve the prognosis and the life quality of patients.

Objective:To investigate the immune effects of Clostridium difficile toxoid B (CdtB) vaccine formulated with different mucosal adjuvants through microneedle immunization, and to provide ideas for the prevention and treatment of Clostridium difficile infection. Methods:CdtB vaccine was prepared with purified Clostridium difficile toxin B(TcdB) after formaldehyde detoxification. Female BALB/c mice were immunized with different doses of vaccine alone or in combination with mucosal adjuvants. The titers of specific serum IgG and fecal IgA were detected at 0 d, 7 d, 14 d, 28 d and 42 d after immunization. The protective effects of CdtB vaccine were evaluated by cell neutralization assay and Clostridium difficile challenge infection. Results:(1) With the increase of immune dose, the mice immunized with CdtB vaccine alone by microneedle not only produced better serum specific IgG, but also had higher level of IgA in feces. (2) When the mice were immunized with CdtB vaccine containing LT or CTB adjuvant by microneedle, the trend of serum specific IgG titer in each group increased with the increase of immune dose, especially in the group containing LT adjuvant. There were significant differences in the trend of specific IgA titer in feces between the adjuvant groups and the group without adjuvant, but the adjuvant effect was not obvious. (3) No significant difference in serum IgG titer was observed between the mice immunized with 10 μg CdtB by microneedle or intraperitoneal injection, but microneedle immunization significantly increased fecal IgA level. (4) The neutralization titers of specific antibodies in mouse serum after immunization and the test results of challenge protection in mice confirmed that the use of CdtB vaccine had certain protective effects.Conclusions:CdtB vaccine had better immune effects in mice through microneedle immunization, but the adjuvant effects of LT and CTB were not significant.

Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously detect the contents of gallic acid, syringin, phellodendrine, aesculin and rhein in the gallnut lotion. Methods An UPLC- MS/MS method was established. Separation was performed on an Agilent Poroshell 120 EC-C₁₈(2.1 mm×150 mm, 2.7 μm)with a gradient mobile phase system of 0.2% formic water-acetonitrile solution. The flow rate was 0.3 ml/min. The temperature of column was 30 ℃. The injection volume was 2 μl. The MS detection was in dynamic MRM mode. Results gallic acid, syringin, phellodendrine, aesculin and rhein were successfully separated using this method, with good linear relationship as the ranges of 153.8-15380、10.31-1031、5.265-526.5、50.70-5070、1.054-105.4 ng/ml, respectively. The precision, repeatability, stability and recovery were good. Conclusion This UPLC-MS/MS method is stable, rapid, and reproducible., It is suitable for detecting the contents of gallic acid, syringin, phellodendrine, esculetin and in the gallnut lotion.

Objective To investigate the change of biological characteristics after stable knockdown of the coding gene of 3-methylcrotonyl-coenzyme A carboxylase β subunit (MCCC2) expression in DU145 by lentivirus shRNA. Methods Three groups were included in this study. shNC was the control group in which MCCC2 was negatively knocked down in DU145. shMCCC2 was the experimental group in which MCCC2 was knocked down. DU145 was the blank group without any treatment. The expression of MCCC2 was assessed by Western blot and qPCR. The proliferation of DU145 cells was detected by CCK8 assay. The migration ability of DU145 was detected by transwell. The apoptosis of DU145 cells was detected by flow cytometry. Results The expression level of MCCC2 in shMCCC2 group was significantly lower than that in shNC group (0.22 ± 0.02 vs 0.61 ± 0.06, P < 0.001). The proliferation (2.24 ± 0.04 vs 3.13 ± 0.15) and migration (23.96 ± 1.85 vs 49.73 ± 0.63) of DU145 cells in shMCCC2 group was significantly lower than that in shNC group, whereas the apoptosis (12.64 ± 0.30 vs 3.68 ± 0.02) of DU145 cells in shMCCC2 was significantly higher than that in shNC group. Conclusion MCCC2 knockdown significantly inhibited the proliferation and migration, and induced apoptosis of DU145 cells, which indicated that the down-regulation of MCCC2 is correlated with the change of tumor biological characteristics of DU145 cell line and can be a potential target for the treatment of prostate cancer.

Angiokinases, such as vascular endothelial-, fibroblast- and platelet-derived growth factor receptors (VEGFRs, FGFRs and PDGFRs) play crucial roles in tumor angiogenesis. Anti-angiogenesis therapy using multi-angiokinase inhibitor has achieved great success in recent years. In this study, we presented the design, synthesis, target identification, molecular mechanism, pharmacodynamics (PD) and pharmacokinetics (PK) research of a novel triple-angiokinase inhibitor WXFL-152. WXFL-152, identified from a series of 4-oxyquinoline derivatives based on a structure-activity relationship study, inhibited the proliferation of vascular endothelial cells (ECs) and pericytes by blocking the angiokinase signals VEGF/VEGFR2, FGF/FGFRs and PDGF/PDGFR simultaneously . Significant anticancer effects of WXFL-152 were confirmed in multiple preclinical tumor xenograft models, including a patient-derived tumor xenograft (PDX) model. Pharmacokinetic studies of WXFL-152 demonstrated high favourable bioavailability with single-dose and continuous multi-dose by oral administration in rats and beagles. In conclusion, WXFL-152, which is currently in phase Ib clinical trials, is a novel and effective triple-angiokinase inhibitor with clear PD and PK in tumor therapy.

Since December 2019, novel coronavirus pneumonia (NCP) has been reported in Wuhan, Hubei Province, and spreads rapidly to all through Hubei Province and even to the whole country. The virus is 2019 novel coronavirus (2019-nCoV), never been seen previously in human, but all the population is generally susceptible. The virus spreads through many ways and is highly infectious, which brings great difficulties to the prevention and control of NCP. Based on the needs of orthopedic trauma patients for emergency surgery and review of the latest NCP diagnosis and treatment strategy and the latest principles and principles of evidence-based medicine in traumatic orthopedics, the authors put forward this expert consensus to systematically standardize the clinical pathway and protective measures of emergency surgery for orthopedic trauma patients during prevention and control of NCP and provide reference for the emergency surgical treatment of orthopedic trauma patients in hospitals at all levels.