The Proctor's reporting guideline for implementation strategies represents a landmark framework in the field of implementation science, aiming to address the issue of inconsistent reporting in implementation research by standardizing the naming, definition, and operationalization of implementation strategies, thereby enhancing the credibility and utility of research findings. This paper provides an in-depth interpretation of the core connotations of this reporting guideline and illustrates its application in developing interview outlines and specifying implementation strategies, using a brief smoking cessation intervention project as a case study. Through this reporting guideline, abstract recommendations for implementation are systematically transformed into clear, multidimensional operational guides, significantly improving the transparency of strategy connotations and the replicability of actual execution. Meanwhile, the case study highlights the flexibility of the guideline, which allows researchers to adapt the content and format of strategies based on local resources and cultural contexts, thus enhancing practical adaptability while maintaining scientific rigor. However, the application of Proctor's reporting guideline still faces challenges, primarily manifested in the potential confusion surrounding the constructs of temporality and dose in practice, as well as the challenges that the inherent flexibility of the guideline may pose to the assessment of fidelity and effectiveness. Despite these limitations, the reporting guideline remains a vital tool for implementation research; future efforts should focus on optimizing its application—through refining operational guidelines, standardizing flexible adaptations, and involving stakeholders—to better guide implementation studies and continuously promote high-quality development in the field.

To explore the advantages of traditional Chinese medicine （TCM） and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications （DMC）， refine key pathophysiological insights and treatment principles， and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine， hosted by the China Association of Chinese Medicine， was held in Beijing， 2024. Experts in TCM， Western medicine， and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis， diagnostic and treatment challenges， and mechanism research related to DMC， ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development， research prioritization， and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM， strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.

AIM:Flow cytometry was used to evaluate the effects of short-term storage conditions(fresh,frozen at-80℃for 7 d,and stored at 4℃for 7 d)on the median fluorescence intensity(MFI)of antibodies and the percentage of immune cell subsets in mouse peripheral blood mononuclear cells(PBMCs)and splenocytes.METHODS:The PBMC and splenocyte suspensions from six male Kunming mice were collected and analyzed under three different processing con-ditions to compare differences in the antibody MFI and percentages of monocyte subsets(Ly-6clow/Ly-6cmedium/Ly-6chigh),macrophages(M1/M2),and dendritic cells.RESULTS:Both tissue and antibody specificity were demonstrated by changes in the antibody MFI values.Following storage at-80℃,the MFIs of certain antibodies(such as CD45 and F4/80 in PBMCs,and CD115,Ly-6c,F4/80,CD80 and MHC-II in the spleen)were similar to those of the fresh groups,where-as after storage at 4℃,the MFIs of other antibodies(such as 7-AAD,CD115,Ly-6c and MHC-II in PBMCs,and CD11b,CD206 and CD11c in the spleen)were closer to those of the fresh groups.The MFI of most of the examined anti-bodies varied significantly following storage.Both storage conditions significantly reduced the viability of PBMCs and sple-nocytes.In PBMCs stored at 4℃,the percentages of total monocytes,Ly-6cmedium/Ly-6chigh monocytes,total macrophages,and dendritic cells were similar to those in the fresh group.Compared with the fresh group,both storage groups presented significantly lower percentages of M1 macrophages and dendritic cells(P<0.05).There were no statistically significant differences in the percentages of total monocytes,Ly-6cmedium monocytes,Ly-6chigh monocytes,total macrophages,M1 and M2 macrophages,or dendritic cells in the spleen among the three groups(P>0.05).The percentage of Ly-6clow monocytes did not differ substantially(P>0.05)between the fresh and-80℃frozen groups but was significantly lower in the 4℃storage group than in the fresh group(P<0.05).CONCLUSION:The storage conditions of the samples had a substantial effect on the flow cytometry results(antibody MFI and cell subset percentages)of the PBMCs and splenic cells,with tissue specificity.If the percentage of immune cell subgroups(particularly monocytes/macrophages/dendritic cells)in PBMCs is highly important,storage at 4℃for 7 d is preferable.If the MFI values of specific antibodies(such as CD45 and F4/80)are important,freezing at-80℃may be more appropriate.If the MFI values of most antibodies or the percentages of criti-cal subgroups(such as total monocytes/Ly-6chigh/total macrophages/dendritic cells)in splenic cells need to be close to those of fresh samples,4 ℃ storage for 7 d is more effective.Freezing at-80℃is preferable if the MFI values of particular anti-bodies(such as CD115 and Ly-6c)need to be determined.

Objective To analyze risk factors of acute cardio-cerebrovascular accidents in elderly patients with lower extremity fracture,and to construct a risk prediction diagram model.Methods A retrospective analysis was conducted on the clinical data of 510 elderly patients with lower extremity fractures,and the patients were divided into the occurrence group and the non-occurrence group based on the occurrence of acute cardiovascular and cerebrovascular accidents during the postoperative hospitalization.Univariate and multivariate Logistic regression analysis was used to analyze risk factors affecting acute cardiovascular and cerebrovascular accidents in elderly patients with lower extremity fracture,and the risk prediction model was constructed and verified.Results A total of 52 cases of acute cardio-cerebrovascular accidents occurred during hospitalization in 510 patients(10.20%).Age,ASA grade Ⅲ ratio,preoperative concurrent cerebrovascular disease,postoperative pneumonia ratio and postoperative bed time≥5 days ratio were higher in the occurrence group than those in the non-occurrence group(P＜0.05).Multivariate Logistic regression analysis showed that age,ASA grade Ⅲ,preoperative concurrent cardiovascular and cerebrovascular disease,and postoperative pneumonia were risk factors for acute cardio-cerebrovascular accidents in elderly patients with lower limb fracture(P＜0.05).Multivariate Logistic regression analysis was used to screen risk factors as predictive variables to build a risk early warning histogram model for acute cardio-cerebrovascular accidents.The total score ranged from 103 to 168 points,corresponding to a risk range of 0.1 to 0.9 points.Receiver operating characteristic curve(ROC)curve results showed that the area under the curve(AUC)of this model for predicting acute cardiovascular and cerebrovascular accidents was 0.980(95%CI:0.964-1.000),and discrete choice approach(DCA)showed that when the risk threshold was 0.2-0.6,the prediction model of this column graph had a good clinical advantage.Conclusion The risk diagram model constructed based on risk factors has good predictive value for the occurrence of acute cardio-cerebrovascular accidents after lower extremity fracture in elderly patients.

Objective To explore the relevant factors affecting the efficacy of traditional Chinese medicine in treating MG,and to develop and validate a nomogram model,in order to personalize the prediction of the probability of benefits for MG patients after traditional Chinese medicine treatment,further guide clinical physicians in targeted medication,and provide guidance for the traditional Chinese medicine treatment of MG.Method Retrospective analysis of MG patients who visited the Affiliated Hospital of Changchun University of Traditional Chinese Medicine from March 2018 to June 2022,13 clinical factors were selected to evaluate the results.To acquire independent factors,univariate and multivariate Logistic analyses were performed.The area under the receiver operating characteristic curve,Harrell's concordance index,calibration curve,and decision curve analyses were used to evaluate the predictive ability,accuracy,and clinical practicability of the prediction model.Results A total of 204 cases were included,divided into derivation cohort 139 cases and temporal validation cohort 65 cases.Multivariate Logistic regression showed 4 independent predictors affecting effectiveness of traditional Chinese medicine in treating MG,including age at onset,repetitive nerve stimulation positive,oral immunosuppressant and anxiety/depression.AUC values for the model group and validation group were 0.76(95%CI:0.68-0.84)and 0.83(95%CI:0.71-0.95),respectively,and based on calibration curve and decision curve analysis,we concluded that the nomogram showed excellent performance.Conclusion MG patients may experience a decrease in short-term efficacy with age,positive RNS,concomitant emotional abnormalities,or recent history of immunosuppressive therapy.This nomogram effectively predicts the possibility of short-term effective traditional Chinese medicine treatment in MG patients.

AIM:To explore suitable methods for the induction and cryopreservation of mouse bone marrow-derived macrophages(BMDMs).METHODS:Mouse fibroblasts(L929 cells)were cultured under varying conditions of temperature,seeding density,and serum concentration.The concentration of macrophage colony-stimulating factor(M-CSF)in the cell supernatant was measured using ELISA to determine the optimal conditions.Mouse bone marrow cells were extracted,and a differential adherence method was employed to pre-culture the bone marrow cells for 4 h,followed by flow cytometry to assess the proportion of resident macrophages.Flow cytometry was utilized to assess the ratio of F4/80 positive cells among the suspended and adherent cells.Induction of BMDMs was conducted using L929 cell supernatant or recombinant M-CSF for 7 d,and flow cytometry was applied to evaluate the proportion of F4/80 and CD11b double-positive cells.The morphologic changes during cell induction were observed under an inverted microscope,and the phagocytic ca-pacity and inflammatory response levels of BMDMs derived from C57BL/6N and C57BL/6J mice were evaluated using neu-tral red and ELISA methods.The cells were immediately cryopreserved after extraction,and then induced after recovery,or cryopreserved after successful induction and recovered.The cell morphology was observed under an inverted micro-scope,cell viability was assessed using the CCK-8 method,and phagocytic ability was measured using the neutral red method.RESULTS:The M-CSF concentration in the supernatant from L929 cells cultured at 33℃,10%fetal bovine se-rum(FBS)for 7 d was rich.Following 4 h of pre-culture,the proportion of F4/80 positive cells in adherent cells was sig-nificantly higher than that in suspended cells(P＜0.01).After 7 d of induction with L929 cell supernatant or recombinant M-CSF,the proportions of F4/80+CD11b+cells showed no significant difference(P＞0.05).Compared with the BMDMs derived from C57BL/6J mice,those from C57BL/6N mice exhibited stronger phagocytic capacity(P＜0.01),and released lower levels of TNF-α(P＜0.01)and IL-6(P＜0.05),and higher levels of IL-1β(P＜0.05).Compared with the BMDMs that were induced after recovery from initial cryopreservation,those cryopreserved immediately after extraction and in-duced upon recovery exhibited better macrophage morphology,higher cell viability(P＜0.01),and enhanced phagocytic ability(P＜0.01).CONCLUSION:The supernatant from L929 cells cultured at 33℃,10%FBS for 7 d is rich in M-CSF,successfully inducing bone marrow cells to differentiate into mouse BMDMs.The differential adherence method for pre-culturing can eliminate resident macrophages from the original bone marrow.The phagocytic capacity and inflammato-ry response levels differ between BMDMs derived from the C57BL/6N and C57BL/6J mouse subtypes.Cryopreserving bone marrow cells immediately after extraction and subsequently inducing them upon recovery is a preferable method for BMDM cryopreservation.

Objective To investigate the correlation between the eosinophil percentage and prognosis in patients with sepsis after intensive care unit(ICU)admission.Methods A retrospec-tive cohort study design was employed,data from adult patients with sepsis or septic shock who were hospitalized for the first time and admitted to the ICU for the first time from the MIMIC-Ⅳ database(version 2.2)were selected.Patients were grouped according to differed eosinophil percentage(＜0.4％and≥0.4％),and the correlation between eosinophil percentage＜0.4％and prognostic out-comes was analyzed.Results A total of 616 patients were included.On the first day after ICU ad-mission,an eosinophil percentage＜0.4％was not significantly associated with prognosis;on the sec-ond day after ICU admission,patients with an eosinophil percentage＜0.4％exhibited a significantly increased 28-day mortality rate(HR=1.572,95％CI,1.119 to 2.208)and in-hospital mortality rate(HR=1.634,95％CI,1.137 to 2.349);on the third day after ICU admission,patients with an eosinophil percentage＜0.4％demonstrated significantly increased 28-day mortality rate(HR=2.072,95％CI,1.482 to 2.896),ICU mortality rate(HR=2.033,95％CI,1.296 to 3.188),and in-hospital mortality rate(HR=2.193,95％CI,1.533 to 3.136).Patients with eosinophil percentages＜0.4％on both the second day and the third day after ICU admission had the worst prog-nosis,with a significantly increased 28-day mortality rate(HR=2.271,95％CI,1.393 to 3.785).Conclusion The eosinophil percentages on the second day and the third day after ICU admission in patients with sepsis are closely correlated with prognosis,particularly aneosinophil percentage＜0.4％on the third day,which is significantly associated with a higher mortality rate.

Objective To observe the effects of electroacupuncture(EA)on morphine-induced microglia activation and analgesic tolerance,and explore the potential mechanism of EA in the treatment of morphine analgesic tolerance.Methods A total of 60 clean-grade SD rats were randomly assigned to control group,morphine group,morphine+EA group,and morphine+EA+colony-stimulating factor 1(CSF1)group,with 15 rats in each group.Morphine analgesic tolerance model was established by continuous 7-d intrathecal injection of morphine in the morphine,morphine+EA and morphine+EA+CSF1 groups.EA was given in the rats of morphine+EA and morphine+EA+CSF1 groups at"Zusanli"and"Sanyinjiao"acupoints,with dilatational wave,frequency of 2/100 Hz,stimulation intensities of 0.5,1.0,and 1.5 mA(10 min per intensity),once a day,for 7 consecutive days.Rats in morphine+EA+CSF1 group were given intrathecal injection with recombinant CSF1 protein for 7 consecutive days.The effect of EA on morphine analgesic tolerance in rats was observed by mechanical withdrawal threshold(MWT).After 7 d,the rats were sacrificed,and the L4-6 spinal dorsal horn and dorsal root ganglion tissues were isolated.The expression of CSF1 protein and mRNA in the dorsal root ganglia and spinal dorsal horn was detected by Western blotting and quantitative polymerase chain reaction.The expression of ionized calcium-binding adapter molecule 1(IBA-1),a marker of microglia in the spinal dorsal horn,was detected by immunofluorescence method,and the expression of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α in the spinal cord was detected by enzyme-linked immunosorbent assay(ELISA).Results After intrathecal injection of morphine,the percentage of maximal possible potential effect(%MPE)in the morphine group was decreased progressively,indicating that the morphine analgesic tolerance model was successfully constructed.Compared with the morphine group,the%MPE in the morphine+EA group was increased after intrathecal injection at 3,5 and 7 d(all P＜0.05).Compared with the morphine+EA group,the%MPE in the morphine+EA+CSF1 group was all decreased after intrathecal injection at 3,5 and 7 d(all P＜0.05).Compared with the control group,the expression of CSF1 protein and mRNA in dorsal root ganglion and the expression of CSF1 protein in spinal dorsal horn in the morphine group were increased(all P＜0.05).Compared with the morphine group,the expression levels of CSF1 protein and mRNA in dorsal root ganglion and CSF1 protein in spinal dorsal horn in the morphine+EA group were decreased(all P＜0.05).There was no significant difference in the expression of CSF1 mRNA in the spinal dorsal horn among those groups(all P＞0.05).Compared with the control group,the expression of IBA-1 in the spinal dorsal horn of the morphine group was increased(P＜0.05).Compared with the morphine group,the expression of IBA-1 in the spinal dorsal horn of the morphine+EA group was decreased(P＜0.05).Compared with the morphine+EA group,the expression of IBA-1 in the spinal dorsal horn of the morphine+EA+CSF1 group was increased(P＜0.05).Conclusion EA can inhibit the activation of microglia in the spinal dorsal horn of rats and improve morphine analgesic tolerance in rats.The mechanism may be related to the reduction of CSF1 protein expression in the spinal dorsal horn.

ObjectiveTo observe the clinical effectiveness and potential mechanism of combined tongue three-needle acupuncture and acupoint application on Lianquan （CV 23） for patients with dysphagia after ischemic stroke. MethodsA prospective study was conducted on 160 patients with post-stroke dysphagia， who were randomly divided into a treatment group and a control group， with 80 cases in each group. The control group received conventional rehabilitation training， while the treatment group received tongue three-needle acupuncture combined with acupoint application on Lianquan （CV 23） on the basis of conventional rehabilitation training， for 4 weeks in both groups. We compared the clinical effectivenss of both groups after treatment， and assessed the swallowing function including videofluoroscopic swallowing study （VFSS）， standardized swallowing assessment （SSA） and functional oral intake scale （FIOS）， swallowing contrast test including hyoid maximum displacement （HmaxD）， pharyngeal transit time （PTT）， and upper esophageal sphincter （UES） opening， surface electromyography （sEMG） test including maximum amplitude and swallowing duration as well as swallowing quality of life questionnaire （SWAL-QOL） score of the patients in both groups before treatment， after 2 weeks and 4 weeks of treatment， respectively. ResultsThe total effective rate in treatment group was 82.50% （66/80）， significantly higher than 66.25% （53/80） in control group （P＜0.05）. The VFSS， and FOIS scores， UES opening rate and HmaxD， sEMG maximal amplitude values， and SWAL-QOL scores were increased in both groups after 2 weeks and 4 weeks of treatment compared with the values before treatment （P＜0.05）， while SSA scores， PTT， and swallowing duration were decreased compared within group before treatment （P＜0.05）. VFSS and FOIS scores， UES opening rate and HmaxD， sEMG maximal amplitude values， and SWAL-QOL scores after 2 and 4 weeks of treatment in the treatment group were higher （P＜0.05）， while SSA scores， PTT， and swallowing duration were lower （P＜0.05） than those in the control group at the same time. ConclusionCombined tongue three-needle acupuncture and acupoint application on Lianquan （CV 23） for patients with dysphagia after ischemic stroke can significantly improve swallowing activities， and its mechanism of action may be related to the improvement of the contraction ability and coordination of swallowing-related muscle groups.

AIM: To investigate the expression of serum growth differentiation factor 11(GDF11)and thrombospondin 1(TSP1)in patients with diabetic retinopathy(DR), and discuss their relationship with microvascular injury.METHODS: Totally 102 DR patients were served as DR group and assigned into non proliferative DR group(NPDR group)and proliferative DR group(PDR group)based on the severity of DR lesions. Meantime, 100 patients with simple diabetes were served as control group. Serum indicators of microvascular injury including vascular endothelial growth factor(VEGF), endothelial cells(ECs), endothelial progenitor cells(EPCs), and levels of GDF11 and TSP1 were measured in each group. Pearson method was used to discuss the correlation between GDF11, TSP1 and microvascular injury indicators. Logistic regression was used to discuss the factors that affected the occurrence of DR. Receiver operating characteristic(ROC)curve was applied to analyze the evaluation value of serum GDF11 and TSP1 for the DR conditions.RESULTS: For the control group, DR group had lower EPCs and GDF11, and higher VEGF, ECs, and TSP1 levels(all P<0.05). The PDR group had lower GDF11 and higher TSP1 than the NPDR group(all P<0.05). Serum GDF11 was negatively related to VEGF and ECs(r=-0.486, -0.511, all P<0.001), and positively related to EPCs(r=0.475, P<0.001). TSP1 was positively related to VEGF and ECs(r=0.579, 0.594, all P<0.001), and negatively related to EPCs(r=-0.505, P<0.001). Moreover, GDF11 and TSP1 were negatively correlated(r=-0.443, P<0.001). The course of T2DM, VEGF, and TSP1 were risk factors for DR, while GDF11 was a protective factor(all P<0.05). The AUC of GDF11, TSP1, and combined diagnosis for PDR conditions was 0.819, 0.822, and 0.915, respectively. The combined diagnosis was better than single diagnosis(Zcombination-GDF11=2.070, P=0.039, Zcombination-TSP1=2.274, P=0.023).CONCLUSION: GDF11 and TSP1 are closely associated with microvascular injury in DR patients and are related to the progression of DR disease, and the combined detection of their serum levels is of clinical value in the assessment of DR disease.