Objective To explore the analgesic initiation mechanism of three-manipulation and three-acupoint tuina in model rats with minor chronic constriction injury(CCI).Methods Fifty-six SD rats were divided randomly into eight groups:normal group,sham group,model 1 group,model 2 group,tuina 1 group,tuina 2 group,tuina 1+transient receptor potential vanilloid-1(TRPV1)antagonist group,and tuina 2+transient receptor potential ankyrin 1(TRPA1)antagonist group.The model,tuina,and tuina+antagonist groups were established with minor CCI models.The tuina and tuina+antagonist groups received the three-method three-point intervention(point method,dial method,kneading method,Yinmen point,Chengshan point,Yanglingquan point)7 days after modeling.The model and sham groups were subjected to grasping restraint,and the normal group received no intervention.After the respective interventions,each group was tested for changes in mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)to detect different types of pain.The nitric oxide(NO)content of the dorsal root ganglion(DRG)was determined by the nitrate reductase method,and changes in protein and gene expression levels of components of the TRPV1/TRPA1-NO-cGMP-protein kinase G(PKG)signaling pathway in the DRG of each group were determined by enzyme-linked immunosorbent assay,Western blot,and qPCR.Results Compared with the model group,MWT and TWL were prolonged in the tuina 1 and tuina 2 groups.Expression levels of TRPV1,TRPA1,NO,soluble guanylate cyclase-β,cGMP,and PKG1 in the DRG were significantly decreased in the tuina 1,tuina 2,tuina 1+TRPV1 antagonist,and tuina 2+TRPA1 antagonist groups.Conclusions Tuina can effectively improve the symptoms of thermal and mechanical hyperalgesia caused by peripheral nerve injury after one-time intervention.Tuina can exert immediate and continuous analgesic effects via the TRPV1/TRPA1-NO-cGMP-PKG signaling pathway.

Unlike chemosynthetic drugs designed for specific molecular and disease targets,active small-molecule natural products typically have a wide range of bioactivities and multiple targets,necessitating extensive screening and development.To address this issue,we propose a strategy for the direct in situ micro-dynamic examination of potential drug candidates to rapidly identify their effects and mechanisms of action.As a proof-of-concept,we investigated the behavior of mussel oligosaccharide(MOS-1)by tracking the subcellular dynamics of fluorescently labeled MOS-1 in cultured cells.We recorded the entire dynamic process of the localization of fluorescein isothiocyanate(FITC)-MOS-1 to the lysosomes and visualized the distribution of the drug within the cell.Remarkably,lysosomes containing FITC-MOS-1 actively recruited lipid droplets,leading to fusion events and increased cellular lipid consumption.These drug behaviors confirmed MOS-1 is a candidate for the treatment of lipid-related diseases.Furthermore,in a high-fat HepG2 cell model and in high-fat diet-fed apolipoprotein E(ApoE)-/-mice,MOS-1 significantly promoted triglyceride degradation,reduced lipid droplet accumulation,lowered serum triglyceride levels,and mitigated liver damage and steatosis.Overall,our work supports the prioritization of in situ visual monitoring of drug location and distribution in subcellular compartments during the drug development phase,as this methodology contributes to the rapid identification of drug indications.Collectively,this methodology is significant for the screening and development of selective small-molecule drugs,and is expected to expedite the identification of candidate molecules with me-dicinal effects.

Objective By observing the effects of"three methods and three points"tuina technique on the expression of interleukin-17F(IL-17F),interleukin-17 receptor C(IL-17RC),activator 1 of nuclear transcription factor-κB(Act1),tumour necrosis factor receptor-associated factor 6(TRAF6)and M1 microglial cell expression in the spinal dorsal horn of rats with mild chronic compressive injury(minor CCI)model,we explored the immediate analgesic mechanism of tuina on peripheral neuropathic pain(pNP).Methods Thirty-six SD rats were divided into the sham group,the model group and the tuina group according to the random number method,twelve rats in each group,and the minor CCI model was replicated by ligating the right sciatic nerve.The rats in the tuina group were subjected to pointing,plucking and kneading at the BL37,BL57 and GB34 points on the affected side using a tuina simulator,while the sham group and the model group were only grasped and restrained,and were intervened for one time.The mechanical pain test and cold plate test were used to evaluate the response of rats to mechanical stimulation and cold stimulation after immediate intervention.The protein expression of IL-17F and TRAF6 in the spinal dorsal horn of rats in each group was detected by Western blotting.The mRNA expression of IL-17F,IL-17RC,Act1 and TRAF6 in the spinal dorsal horn of rats in each group was detected by real-time PCR.The average fluorescence intensity of M1 microglia in the spinal dorsal horn of rats in each group was detected by immunofluorescence.Results Behavioral results showed that before intervention,compared with the sham group,paw mechanical withdraw threshold(PMWT)decreased and cold sensitivity threshold(CST)increased in the model group and the tuina group;after tuina intervention,PMWT in the tuina group was increased,and CST was decreased compared with the model group;after intervention,PMWT in the tuina group was increased,while CST was decreased(P＜0.05).RT-PCR results showed that compared with the sham group,mRNA expression levels of IL-17F,IL-17RC,TRAF6 and Act1 in the spinal dorsal horn of the model group were increased;compared with model group,the mRNA expression levels of above indexes in the tuina group were decreased(P＜0.05).Western boltting results showed that compared with the sham group,the expression levels of IL-17F and TRAF6 protein in the spinal dorsal horn of the model group were increased;compared with the model group,the expression levels of IL-17F and TRAF6 protein in the tuina group decreased(P＜O.05).Immunofluorescence results showed that the mean fluorescence intensity of CD40 in the spinal dorsal horn of model group was enhanced compared with the sham group;compared with the model group,the mean fluorescence intensity of CD40 in the tuina group was decreased(P＜0.05).Conclusion The"three methods and three points"tuina technique can produce immediate analgesia by inhibiting the expression of IL-17F,IL-17RC,Act1,TRAF6 and the activation of M1 microglia in the dorsal horn of the spinal cord after one intervention.

Objective To explore the analgesic initiation mechanism of"three-manipulations and three-acupoints"of tuina on minor chronic constriction injury(minor CCI)model rats.Methods According to the random number table method,35 SD rats were randomly divided into five groups:normal group,sham group,model group,tuina group,and tuina+MK-801 group.The model group,tuina group,and tuina+MK-801 group were subjected to ligation of the right sciatic nerve trunk to establish a minor CCI rat model.The sham group was only exposed to the right sciatic nerve without ligation,and the normal group was not subjected to any operation.The normal group was not subjected to any intervention measures.On the seventh day after modeling,the model group and the sham group underwent 9 minutes of grasping restraint,while the tuina group underwent one intervention of three-manipulations(point method,dialing method,and kneading method)and three-acupoints(right"Yinmen"(BL37),"Chengshan"(BL57),and"Yanglingquan"(GB34)acupoints)with each manipulation and acupoint intervention for 1 minute for a total of 9 minutes.The tuina+MK-801 group received intrathecal injection of MK-801 from the fifth to seventh days after modeling,with a dose of 6 μg(10 μL)per day,tuina intervention was performed 30 minutes after the last intrathecal injection,and the specific operation of tuina was the same as that of the tuina group.Before modeling,after modeling,and after intervention,each group of rats was subjected to cold sensitivity threshold(CST)and mechanical withdrawal threshold(MWT)testing.After intervention,immunohistochemistry was used to detect the positive expression of cyclic guanosine monophosphate(cGMP)in the spinal dorsal horn(SDH)at L4-6 segments;protein expressions of N-methyl-D-aspartate receptor 1(NMDAR1),neurogenic nitric oxide synthase(nNOS),soluble guanylyl cyclase β(sGCβ),and protein kinase G1(PKG1)in SDH at L4-6 segments were detected by Western blotting;mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 in SDH at L4-6 segments were detected by real-time PCR.Results Compared with the normal and sham groups,after modeling,CST increased and MWT decreased in the model group,tuina group and tuina+MK-801 group(P＜0.05);after intervention,the positive protein expression of cGMP was increased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were increased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were increased in SDH at L4-6 segments in the model group(P＜0.05).Compared with the model group,after intervention,CST decreased and MWT increased in the tuina group and tuina+MK-801 group(P＜0.05);the positive protein expression of cGMP was decreased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were decreased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were decreased in SDH at L4-6 segments in the tuina group and tuina+MK-801 group(P＜0.05).Conclusion One-time tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia induced by peripheral nerve injury,which may initiate analgesia through the NMDAR1/cGMP/protein kinase G signaling pathway,thereby exerting immediate analgesic effect.

Objective To develop an ultra-performance liquid chromatography-mass spectrometry(UPLC-MS/MS)method for the simultaneous quantification of dolutegravir,raltegravir,efavirenz,lamivudine and tenofovir in human plasma and to apply it to the therapeutic monitoring.Methods Dolutegravir-D5,raltegravir-D4,efavirenz-D5,lamivudine-13 C-15 N2 and tenofovir-D7 were used as internal standard,respectively.All samples were extracted using the protein precipitation method with acetonitrile and then diluted for analysis.Chromatographic separation was performed on Shim-pack XR-ODS Ⅲ(2.0 mmx50 mm,1.6 μm)column.Mobile phases A and B consisted of 0.1％formic acid in water and acetonitrile respectively.A programmed mobile phase gradient was used at a flow rate of 0.3 mL·min-1 and column temperature of 40 ℃.The tandem mass spectrometer was equipped with an electrospray ionization(ESI)source operating in multiple reaction monitoring(MRM)modes.After methodological validation,it can be used for therapeutic drug monitoring in HIV patients.Results There was good linearity in the validated concentration ranges of 62.5-3 000 ng·mL-1 for dolutegravir,10-500 ng·mL-1 for raltegravir,125-6 000 ng·mL-1for efavirenz,10-500 ng·mL-1 for lamivudine and 10-500 ng·mL-1 for tenofovir with the linear correlation coeffificients of determination(R2)of all higher than 0.998.The accuracy of both intra-day and inter-day studies ranged from 94.0％-109.3％,and the relative standard deviations were less than 7％.The IS-normalized matrix factor and extraction recoveries of all analytes were 95.7％-106.0％and 98.7％-104.5％at all concentrations.All analytes were stable in plasma at a certain storage environment.The trough blood concentrations of dolutegravir,efavirenz,lamivudine and tenofovir were 107.7-2 366.0,740.0-3 410.0,38.5-1 229.3,31.6-224.4ng·mL-1 in HIV patients,respectively.Conclusion The method is highly aceurate,easy to perform,low-cost,and suitable for therapeutic drug monitoring of dolutegravir,raltegravir,efavirenz,lamivudine and tenofovir in HIV patients.

Objective:To analyze the clinical characteristics of chronic Q fever.Methods:The epidemiological characteristics, clinical manifestations, laboratory results, diagnosis, treatment and prognosis of 12 patients with chronic Q fever admitted in Peking Union Medical College Hospital from January 2008 to December 2022 were analyzed retrospectively.Results:Of the12 cases, there were 11 males and 1 female with a median age at diagnosis of 58 years (47-68). The median time interval between the onset of symptoms and diagnosis was 9.5 months (4.0-28.5). Ten patients had fever, and the common symptoms included weakness ( n=6), weight loss ( n=5) and shortness of breath ( n=3). Seven patients presented with hepatomegaly and splenomegaly. Among the 12 patients, 1 patient suffered from cardiac valve and artery involvement at the same time, 8 patients suffered from infective endocarditis, and 1 patient suffered from aneurysm. No definite infection site was found in 2 patients. Seven of the 12 patients (58.3%) had elevated serum creatinine. Rheumatoid factors were detected in 8 patients, all of which were elevated. Nine patients were treated with doxycycline/minocycline plus hydroxylchloroquine and 3 patients were treated with doxycycline/minocycline. Patients were followed up for 3 months to 14 years, 7 patients stopped antibiotics after 12 to 30 months and were in stable condition. One patient died of heart failure 6 months after diagnosis and 1 patient lost follow-up. Three patients diagnosed in 2022 were still under treatment and in stable condition. Conclusions:The clinical manifestations of chronic Q fever lack of specificity and may involve multiple systems. Endocarditis and mycotic aneurysm are common complications. For long-term fever with new heart valve function damage or mycotic aneurysm patients, chronic Q fever should therefore be considered.

Nonpharmaceutical interventions (NPIs) have been commonly deployed to prevent and control the spread of the coronavirus disease 2019 (COVID-19), resulting in a worldwide decline in influenza prevalence. However, the influenza risk in China warrants cautious assessment. We conducted a cross-sectional, seroepidemiological study in Shandong Province, Northern China in mid-2021. Hemagglutination inhibition was performed to test antibodies against four influenza vaccine strains. A combination of descriptive and meta-analyses was adopted to compare the seroprevalence of influenza antibodies before and during the COVID-19 pandemic. The overall seroprevalence values against A/H1N1pdm09, A/H3N2, B/Victoria, and B/Yamagata were 17.8% (95% CI 16.2%-19.5%), 23.5% (95% CI 21.7%-25.4%), 7.6% (95% CI 6.6%-8.7%), and 15.0 (95% CI 13.5%-16.5%), respectively, in the study period. The overall vaccination rate was extremely low (2.6%). Our results revealed that antibody titers in vaccinated participants were significantly higher than those in unvaccinated individuals (P < 0.001). Notably, the meta-analysis showed that antibodies against A/H1N1pdm09 and A/H3N2 were significantly low in adults after the COVID-19 pandemic (P < 0.01). Increasing vaccination rates and maintaining NPIs are recommended to prevent an elevated influenza risk in China.

Objective:The surgical method and clinical efficacy was studied on the anterior lateral malleolar venous network grafting in repair of the defects of superficial palmar arch.Methods:From February 2015 to July 2021, 8 cases (6 males and 2 females; aged 32-44 years old, mean age at 39 years old) were treated in the Department of Hand Surgery of Yantaishan Hospital. Anterior lateral malleolar venous network was grafted to repair the defects of superficial palmar arch of the patients. The patients had different degrees of tendon damage, and injuries of nerve, deep palmar arch, common palmar digital artery or proper palmar digital artery. The length of the vessel defect was 6.0-10.0 cm. Blood supply insufficiency occurred in 1st-5th digits in 1 case, 2nd-4th fingers in 5 cases, and 2nd-5th fingers in 2 cases. The anterior lateral malleolar venous network was transferred to repair 3 broken ends of vessels in 5 cases, 4 broken ends in 1 case and 5 broken ends in 2 cases. The wound of hand was sutured directly in 2 cases. The emergency surgical repair of the wound of hand by transfer of ALTPF was performed in 2 patients, and elective ALTPF transfer in 3 cases. Forearm dorsi-ulnar flap transfer was performed in 1 case. The follow-up reviews were carried out via the outpatient clinic visit, telephone, WeChat interviews or home visits. The blood supply of the distal digit, skin sensation, TPD, the range of motion of the phalangeal joints were observed.Results:The results showed that all the digits and the grafted flaps survived. The follow-up was carried out for 10-53 months, with an average of 17.9 months. The functions of digit extension and flexion, finger opposition and thumb opposition were restored in 5 cases, and the patients returned back to former jobs. The finger flexion and thumb opposition were limited in 3 cases, and these cases participated in light physical work. The proprioception of the digits were recovered, and the TPD was 5-7 mm. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association and rated as excellent in 5 cases and good in 3 cases. The donor sites of the foot showed a linear scar. There was no effect on the function and the shape of the donor site.Conclusion:The anterior lateral malleolar venous network grafting is an ideal method in repair of the defects of superficial palmar arch.

Objective:To explore the clinical application of the posterior tibial artery perforator flap with the great saphenous vein (GSV) in the treatment of severe degloving injury of the forearm.Methods:From June 2015 to October 2020, 5 patinets (4 males and 1 female, aged 20-46 years old, mean age 37 years old) were treated in the Department of Hand Surgery of Yantaishan Hospital in Yantai. Aposterior tibial artery perforator flap with GSV was used to repair the partial wound of the injured forearm, together with the establishment of venous circulation of dorsal hand for all 5 patients. All the patients suffered from severe forearm degloving injury. Of which, 3 accompanied with ulna radius fracture, 2 with ulnar and radial artery injury and 2 with blood supply insufficiency in the injured fingers. The sizes of soft-tissue defect were 26 cm×18 cm-32 cm×25 cm. The sizes of the posterior tibial artery perforator flap with GSV ranged from 12 cm×5 cm to 33 cm×6 cm. The anastomoses were performed on the perforating artery and the radial artery. The GSV was anastomosed with cephalic vein with bridging anastomosis to re-establish the venous circulation of dorsal hand. Combined anterolateral thigh flaps (ALTF) were used to repair the rest wound of injured limbs in 4 patients, and the combined pedicled abdominal flap was used in 1 patient. The donor site of calf flap was sutured directly, and the skin of upper limb was thinned into medium thick to cover the thigh flap donor site, which was packed with pressure dressing. Regular follow-up reviews were carried out by outpatient clinic, telephone, WeChat APP or home-visit after the operation to observe the survival of flaps, the swelling of the distal end of injured limb, functional recovery and healing at the donor site. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:All the grafted flaps in 5 patients survived. Necrosis at epidermal edge occurred in 1 calf flap, and achieved secondary healing after dress changing for 4 weeks. There was neither arterial nor venous crisis in all flaps. The oedema of the hand disappeared within 4-8 weeks, with clear dermatoglyph. There was no obvious enlarged circumference of thumb and fingers. The follow-up was carried out for 8-20 months, with 11 months in average. The flaps were elastic in good colour, and full texture without pigmentation. The donor site of the calf flap showed a linear scar. No swollen was seen in the hands of the injured forearm and the feet of donor lower legs. The range of motion of phalangeal joints was good. Recovery of finger sensation achieved at S 4. The recovery of the sensation of posterior tibial artery perforator flap reached S 3 in 2 patients and S 2 in 3 patients. Assessment of the Upper Limb Function using the Standard Issued by the Hand Surgery Society of Chinese Medical Association rated excellent in 3 patients and good in 2 patients. Conclusion:The posterior tibial artery perforator flap with GSV has a reliable blood supply with a small damage to the donor site. This flap is ideal for repair of severe degloving injury of forearm and meanwhile to achieve the re-establishment of venous circulation in dorsal hand.

Hepatocellular carcinoma (HCC) has been known as the second common leading cancer worldwide, as it responds poorly to both chemotherapy and medication. Triptolide (TP), a diterpenoid triepoxide, is a promising treatment agent for its effective anticancer effect on multiple cancers including HCC. However, its clinical application has been limited owing to its severe systemic toxicities, low solubility, and fast elimination in the body. Therefore, to overcome the above obstacles, photo-activatable liposomes (LP) integrated with both photosensitizer Ce6 and chemotherapeutic drug TP (TP/Ce6-LP) was designed in the pursuit of controlled drug release and synergetic photodynamic therapy in HCC therapy. The TP encapsulated in liposomes accumulated to the tumor site due to the enhanced permeability and retention (EPR) effect. Under laser irradiation, the photosensitizer Ce6 generated reactive oxygen species (ROS) and further oxidized the unsaturated phospholipids. In this way, the liposomes were destroyed to release TP. TP/Ce6-LP with NIR laser irradiation (TP/Ce6-LP+L) showed the best anti-tumor effect both