ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations， to evaluate the in vitro anticoagulant， antiplatelet and fibrinolytic activities of the three preparations， and to investigate the effects of temperature， pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay（BCA） and Bradford method， and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time（Fibg-TT） method， and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time（APTT）， prothrombin time（PT） and thrombin time（TT）. The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature， pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage， the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa， the bands of Maixuekang capsules were more clear in the range of >25 kDa， and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa， primarily containing peptides below 2 kDa， most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage， The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration， the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate， whereas for Maixuekang capsules， the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage， the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills， the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules， and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate（ADP） and arachidonic acid（AA）， and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules（P<0.01）， whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating， while the activities of the two capsules decreased significantly within 5 min above 80 ℃， and continued to decrease within 2 h. Compared with pure water， the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity（pH 1-3）. In the pepsin buffer， the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times， while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased， the lowest levels were about 60% and 20%， respectively. In trypsin buffer， the anticoagulant activities of Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules decreased significantly， and the lowest levels decreased to about 41%， 41% and 35%， respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder， dried Hirudo powder and reflux extract of Hirudo decrease sequentially， and the anticoagulant activity decrease gradually， but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively， and the first two are suitable for enteric-soluble preparations， while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods， active substances， pharmacodynamics and mechanism of Hirudo preparations.

ObjectiveTo evaluate the effectiveness of stone needle thermocompression and massage compared to flurbiprofen gel patch in relieving chronic musculoskeletal pain in the shoulder and back， and to explore the potential mediating mechanism through muscle elasticity. MethodsA total of 120 patients with chronic musculoskeletal pain in the shoulder and back were randomly assigned to either stone needle group or flurbiprofen group， with 60 patients in each. The stone needle group received stone needle thermocompression and massage for 30 minutes， three times per week； the flurbiprofen group received flurbiprofen gel patch twice daily. Both groups were treated for 2 weeks. Pain improvement， as the primary outcome， was assessed using the Global Pain Scale （GPS） at baseline， after 2 weeks of treatment， and again 2 weeks post-treatment. To explore potential mechanisms， a mediator analysis was conducted by measuring changes in superficial and deep muscle elasticity using musculoskeletal ultrasound at baseline and after the 2-week treatment period. ResultsThe stone needle group showed significantly greater pain relief than the flurbiprofen group 2 weeks post-treatment. After adjusting for confounders related to pain duration， the between-group mean difference was -8.8 ［95% CI （-18.2， -0.7）， P＜0.05］. Part of the therapeutic effect was mediated by changes in deep muscle elasticity， with a mediation effect size of -1.5 ［95% CI （-2.0， -0.9）， P = 0.024］， accounting for 17.9% of the total effect. ConclusionStone needle thermocompression and massage can effectively relieve chronic musculoskeletal pain in the shoulder and back， partly through a mediating effect of improved deep muscle elasticity.

Objective: To investigate the objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease (DKD). Methods: A cross-sectional study enrolled 134 patients with DKD G3-5 stages who met the diagnostic criteria for damp-heat syndrome in DKD. The patients were treated at Dongzhimen Hospital, Beijing University of Chinese Medicine, from May 2023 to January 2024. The patients were divided into three groups: DKD G3, DKD G4, and DKD G5 stage, with 53, 33, and 48 patients in each group, respectively. Clinical general data (gender, age, and body mass index) and damp-heat syndrome scores were collected from the patients. The YZAI-02 traditional Chinese medicine (TCM) AI Tongue Image Acquisition Device was used to capture tongue images from these patients. The accompanying AI Open Platform for TCM Tongue Diagnosis of the device was used to analyze and extract tongue manifestation features, including objective data on tongue color, tongue quality, coating color, and coating texture. Clinical data and objective tongue manifestation characteristics were compared among patients with DKD G3-5 based on their DKD damp-heat syndrome status. Results: No statistically significant difference in gender or body mass index was observed among the three patient groups. The DKD G3 stage group had the highest age (P<0.05). The DKD G3 stage group had a lower score for symptoms of poor appetite and anorexia(P<0.05) than the DKD G5 group. No statistically significant difference was observed in damp-heat syndrome scores among the three groups. Compared with the DKD G5 stage group, the DKD G3 stage group showed a decreased proportion of pale color at the tip and edges of the tongue (P<0.05). The DKD G4 stage group exhibited an increased proportion of crimson at the root of the tongue, a decreased proportion of thick white tongue coating at the root, a decreased proportion of pale color at the tip and edges of the tongue, an increased hue value (indicating color tone) of the tongue color in the middle, an increased brightness value (indicating color lightness) of the tongue coating color in the middle, and an increased thickness of the tongue coating (P<0.05). No statistically significant difference was observed in other tongue color proportions, color chroma values, body characteristics, coating color proportions, coating color chroma values, and coating texture characteristics among the three groups. Conclusion Tongue features differ in different stages of DKD damp-heat syndrome in multiple dimensions, enabling the inference that during the DKD G5 stage, the degree of qi and blood deficiency in the kidneys, heart, lungs, liver, gallbladder, spleen, and stomach is prominent. Dampness is more likely to accumulate in the lower jiao, particularly in the kidneys, whereas heat evil in the spleen and stomach is the most severe. These insights provide novel ideas for the clinical treatment of DKD.

Depression is a common chronic mental illness.Animal models of depression are widely used to study the pathogenesis of depression as well as in the development of new antidepressant drugs.The consistency and reliability of animal models of depression to simulate human depressive symptoms directly affect the study result.However,at present,no animal models have been found that are completely consistent with the onset of human depression,which hinders the in-depth study of depression.By using scientific and reasonable modeling and evaluation method,the pathological state of depression can still be simulated to a large extent,providing a basis for further in-depth research of the pathogenesis of depression and the development of effective antidepressant drugs.This paper aims to provide a reference for researchers by reviewing several commonly used animal models and behavioral tests of depression.

Objective To investigate the effect of electroacupuncture(EA)on the behaviors of lipopolysaccharide(LPS)-induced depression-like mice and explore the potential molecular mechanism of the antidepressant effect of EA using transcriptomics sequencing technology.Methods Thirty-six male specific pathogen-free-grade C57BL/6J mice were divided into the normal,the model,and the EA groups using the random number table method,with 12 mice per group.The EA group received a daily 20-minute EA at the"Baihui"and"Yintang"acupoints for 7 days.After 7 days of EA treatment,1.5 mg/kg LPS was injected intraperitoneally into the model and EA groups,whereas the normal group received an injection of an equal volume of saline solution.Depressive behaviors were assessed using the open-field test,tail-suspension test,and other relevant tests.Pathologic morphology of the hippocampus tissue was determined using hematoxylin-eosin staining.Transcriptomics technology was used to identify differentially expressed genes in the hippocampus,and bioinformatics analysis was performed.Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of hematopoietic cell kinase(Hck),adhesion G protein-coupled receptor E1(Adgre1),Toll-like receptor 2(Tlr2),and Gm49037 in the hippocampus.Results Compared to the normal group,mice in the model group exhibited a decrease in total travel distance,central travel distance,center area time(P＜0.01),as well as an increase in tail-suspension immobility time(P＜0.05).Hippocampal neurons were loosely arranged,and some neurons had blurred boundaries.Compared to the model group,mice in the EA group showed an increase in open-field total travel distance,central travel distance,center area time(P＜0.05),as well as a decrease in tail-suspension immobility time(P＜0.01);Additionally,hippocampal neurons were more tightly arranged,exhibiting clearer morphology and structure.Transcriptomic analysis revealed that 779 differentially expressed genes were identified in the model group compared to the normal group,whereas 172 genes differed between the EA and model groups.Nineteen genes were upregulated in the model group but downregulated in the EA group,and ten genes were down-regulated in the model group but up-regulated in the EA group.Gene Ontology Functional Analysis indicated that these genes primarily function in immune responses,Toll-like receptor activity,cell surface receptor signaling pathways,and interleukin-10 production.Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed enrichment in inflammation-related pathways,including Toll-like receptor signaling pathway and tumor necrosis factor signaling pathway.Compared to the normal group,Hck,Adgre1,and Tlr2 mRNA expressions were elevated in the model group(P＜0.01),whereas there was a decrease tendency for Gm49037 mRNA expression and there was no significant difference(P＞0.05);compared to the model group,Hck,Adgre1,and Tlr2 mRNA expressions were decreased in the EA group(P＜0.05),whereas there was an increase tendency for Gm49037 mRNA expression and there was no significant difference(P＞0.05).Conclusion EA alleviated depressive-like behavior and pathological damage in the hippocampus in LPS-induced mice,which may be related to attenuating hippocampal inflammatory pathways,and Hck,Adgre1,and Tlr2 genes may be the potential targets for its effect.

Objective To investigate the effect of electroacupuncture(EA)on the behaviors of lipopolysaccharide(LPS)-induced depression-like mice and explore the potential molecular mechanism of the antidepressant effect of EA using transcriptomics sequencing technology.Methods Thirty-six male specific pathogen-free-grade C57BL/6J mice were divided into the normal,the model,and the EA groups using the random number table method,with 12 mice per group.The EA group received a daily 20-minute EA at the"Baihui"and"Yintang"acupoints for 7 days.After 7 days of EA treatment,1.5 mg/kg LPS was injected intraperitoneally into the model and EA groups,whereas the normal group received an injection of an equal volume of saline solution.Depressive behaviors were assessed using the open-field test,tail-suspension test,and other relevant tests.Pathologic morphology of the hippocampus tissue was determined using hematoxylin-eosin staining.Transcriptomics technology was used to identify differentially expressed genes in the hippocampus,and bioinformatics analysis was performed.Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of hematopoietic cell kinase(Hck),adhesion G protein-coupled receptor E1(Adgre1),Toll-like receptor 2(Tlr2),and Gm49037 in the hippocampus.Results Compared to the normal group,mice in the model group exhibited a decrease in total travel distance,central travel distance,center area time(P＜0.01),as well as an increase in tail-suspension immobility time(P＜0.05).Hippocampal neurons were loosely arranged,and some neurons had blurred boundaries.Compared to the model group,mice in the EA group showed an increase in open-field total travel distance,central travel distance,center area time(P＜0.05),as well as a decrease in tail-suspension immobility time(P＜0.01);Additionally,hippocampal neurons were more tightly arranged,exhibiting clearer morphology and structure.Transcriptomic analysis revealed that 779 differentially expressed genes were identified in the model group compared to the normal group,whereas 172 genes differed between the EA and model groups.Nineteen genes were upregulated in the model group but downregulated in the EA group,and ten genes were down-regulated in the model group but up-regulated in the EA group.Gene Ontology Functional Analysis indicated that these genes primarily function in immune responses,Toll-like receptor activity,cell surface receptor signaling pathways,and interleukin-10 production.Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed enrichment in inflammation-related pathways,including Toll-like receptor signaling pathway and tumor necrosis factor signaling pathway.Compared to the normal group,Hck,Adgre1,and Tlr2 mRNA expressions were elevated in the model group(P＜0.01),whereas there was a decrease tendency for Gm49037 mRNA expression and there was no significant difference(P＞0.05);compared to the model group,Hck,Adgre1,and Tlr2 mRNA expressions were decreased in the EA group(P＜0.05),whereas there was an increase tendency for Gm49037 mRNA expression and there was no significant difference(P＞0.05).Conclusion EA alleviated depressive-like behavior and pathological damage in the hippocampus in LPS-induced mice,which may be related to attenuating hippocampal inflammatory pathways,and Hck,Adgre1,and Tlr2 genes may be the potential targets for its effect.

Objective:To evaluate the effect of different psychosocial interventions on fertility concerns in cancer patients using network Meta-analysis.Methods:Randomized controlled trials or quasi-experimental studies on the effects of different psychosocial interventions on fertility concerns in cancer patients were electronically searched in China National Knowledge Infrastructure, Wanfang Data, VIP, China Biology Medicine disc, PubMed, Embase, Cochrane Library, CINAHL, Web of Science and PsycINFO. The search period was from database establishment to October 31, 2024. Two researchers independently conducted literature screening and data extraction. Conventional Meta-analysis of the included literature was performed using RevMan 5.4, and network Meta-analysis was performed using Stata 18.0.Results:A total of 10 studies involving 835 cancer patients and seven psychosocial interventions were included. Network Meta-analysis showed that there was a statistically significant difference in the intervention effect of fertility counseling ( MD=18.24, 95% CI: 2.85-33.62, P<0.05) and mental awareness training ( MD=17.97, 95% CI: 3.09-32.85, P<0.05) on fertility concerns in cancer patients as compared to routine nursing. The ranked probability plot showed that fertility counseling and mental awareness training had superior results. Conclusions:Fertility counseling has a superior effect on fertility concerns in cancer patients. Mental awareness training may be effective in improving fertility concerns in cancer patients, and further research is needed to validate the effectiveness of mindfulness-based stress reduction, cognitive behavioral therapy, dyadic coping interventions, and educational interventions in improving fertility concerns in cancer patients.

Depression is a common chronic mental illness.Animal models of depression are widely used to study the pathogenesis of depression as well as in the development of new antidepressant drugs.The consistency and reliability of animal models of depression to simulate human depressive symptoms directly affect the study result.However,at present,no animal models have been found that are completely consistent with the onset of human depression,which hinders the in-depth study of depression.By using scientific and reasonable modeling and evaluation method,the pathological state of depression can still be simulated to a large extent,providing a basis for further in-depth research of the pathogenesis of depression and the development of effective antidepressant drugs.This paper aims to provide a reference for researchers by reviewing several commonly used animal models and behavioral tests of depression.

Objective:To evaluate the effect of different psychosocial interventions on fertility concerns in cancer patients using network Meta-analysis.Methods:Randomized controlled trials or quasi-experimental studies on the effects of different psychosocial interventions on fertility concerns in cancer patients were electronically searched in China National Knowledge Infrastructure, Wanfang Data, VIP, China Biology Medicine disc, PubMed, Embase, Cochrane Library, CINAHL, Web of Science and PsycINFO. The search period was from database establishment to October 31, 2024. Two researchers independently conducted literature screening and data extraction. Conventional Meta-analysis of the included literature was performed using RevMan 5.4, and network Meta-analysis was performed using Stata 18.0.Results:A total of 10 studies involving 835 cancer patients and seven psychosocial interventions were included. Network Meta-analysis showed that there was a statistically significant difference in the intervention effect of fertility counseling ( MD=18.24, 95% CI: 2.85-33.62, P<0.05) and mental awareness training ( MD=17.97, 95% CI: 3.09-32.85, P<0.05) on fertility concerns in cancer patients as compared to routine nursing. The ranked probability plot showed that fertility counseling and mental awareness training had superior results. Conclusions:Fertility counseling has a superior effect on fertility concerns in cancer patients. Mental awareness training may be effective in improving fertility concerns in cancer patients, and further research is needed to validate the effectiveness of mindfulness-based stress reduction, cognitive behavioral therapy, dyadic coping interventions, and educational interventions in improving fertility concerns in cancer patients.