OBJECTIVES: To explore the therapeutic effect of LuoFuShan Rheumatism Plaster (LFS) on neuropathic pain (NP) and its molecular mechanism. METHODS: Mouse models of sciatic nerve chronic constriction injury (CCI) were treated with low, medium, and high doses (2.2, 4.4, and 8.8 cm², respectively) of LFS by topical application for 14 consecutive days. The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold (MWT), paw withdrawal latency (PWL), plasma IL-6 and TNF-α levels, and histopathology of the sciatic nerve. Network pharmacology and molecular docking were used to identify the key targets and signaling pathways. The key targets were verified by RT-qPCR and immunohistochemistry. The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart, liver, and kidneys. RESULTS: Compared with the CCI group, LFS dose-dependently increased MWT and PWL, reduced plasma IL-6 and TNF-α levels, and alleviated sciatic nerve inflammation in the mouse models. Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling. Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF‑α. In the mouse models of sciatic nerve CCI, LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve. LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart, liver and kidneys as compared with those in the CCI model group and sham-operated group. CONCLUSIONS LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.
Animals ; Toll-Like Receptor 4/metabolism* ; Neuralgia/metabolism* ; Mice ; Signal Transduction/drug effects* ; Tumor Necrosis Factor-alpha/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Sciatic Nerve/injuries* ; Male ; Molecular Docking Simulation ; Disease Models, Animal ; Interleukin-6

Objective To explore the therapeutic effect of LuoFuShan Rheumatism Plaster(LFS)on neuropathic pain(NP)and its molecular mechanism.Methods Mouse models of sciatic nerve chronic constriction injury(CCI)were treated with low,medium,and high doses(2.2,4.4,and 8.8 cm2,respectively)of LFS by topical application for 14 consecutive days.The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold(MWT),paw withdrawal latency(PWL),plasma IL-6 and TNF-α levels,and histopathology of the sciatic nerve.Network pharmacology and molecular docking were used to identify the key targets and signaling pathways.The key targets were verified by RT-qPCR and immunohistochemistry.The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart,liver,and kidneys.Results Compared with the CCI group,LFS dose-dependently increased MWT and PWL,reduced plasma IL-6 and TNF-α levels,and alleviated sciatic nerve inflammation in the mouse models.Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling.Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF-α.In the mouse models of sciatic nerve CCI,LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve.LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart,liver and kidneys as compared with those in the CCI model group and sham-operated group.Conclusion LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.

Objective:To investigate the current status of subjective well-being among stroke patients, and to explore the pathways and effects of influencing factors using structural equation model, so as to provide reference for improving subjective well-being among stroke patients.Methods:From July to November 2024, the stroke patients admitted to the First Affiliated Hospital of Bengbu Medical University, the Second Affiliated Hospital of Bengbu Medical University, Hefei First People′s Hospital were selected by convenience sampling method. A cross-sectional survey was conducted using a general demographic questionnaire, General Well-Being Scale, Cognitive Reserve Index questionnaire, Social Support Rating Scale, Stroke Symptom Cluster Scale, and FRAIL Scale, and AMOS 26.0 was used to analyse the pathways and effects of influencing factors of subjective well-being.Results:A total of 435 questionnaires were collected, 410 were valid.Among 410 cases, 266 case were males, 144 were females, with an age of (65.96 ± 12.15) years. The subjective well-being scores of stroke patients were (72.58 ± 11.66) points. Cognitive reserve and social support were positively correlated with subjective well-being ( r = 0.517, 0.554, both P<0.01), while symptom burden and frailty were negatively correlated with subjective well-being ( r = -0.687, -0.670, both P<0.01). Path analysis showed that symptom burden, frailty, cognitive reserve, and social support had a direct impact on subjective well-being (path coefficients were -0.500, -0.266, 0.148, and 0.144, respectively, all P<0.05), while cognitive reserve, social support, and symptom burden had an indirect impact on subjective well-being (path coefficients were 0.287, 0.249, and 0.108, respectively, all P<0.05). Conclusions:The subjective well-being of stroke patients is influenced by multiple factors, with symptom burden being an important factor affecting subjective well-being. Intervention strategies such as improving cognitive reserve, strengthening social support systems, and preventing frailty can improve the subjective well-being of patients.

Objective To evaluate the clinical effect of intensive treatment of obese patients with type 2 diabetes with Degu asparagus insulin and semaglutide.Methods A total of 92 obese patients with type 2 diabetes who were admitted to Rugao Hospital of Traditional Chinese Medicine from May 2020 to May 2023 were selected and randomly divided into two groups by randomnumber table method,with 46 cases in each group.The treatment group received Degu asparagus insulin and semaglutide,and the control group was treated with semaglutide.Glucose and lipid metabolism indicators(fasting blood glucose,glycated hemoglobin,fructosamine,2-hour postprandial blood glucose,and total cholesterol),blood glucose fluctuations(standard deviation of blood glucose,amplitude of postprandial blood glucose fluctuations,24-hour average blood glucose),insulin resistance index,visceral fat index,antioxidant indicators(malondialdehyde[MDA],lipid peroxide[LPO],superoxide dismutase[SOD],and paraoxonase-1[PON1]),and adverse reactions were observed before and after 4 weeks of treatment.Results After 4 weeks of treatment,the treatment group showed a significant improvement in glucose and lipid metabolism compared to the control group,including decreased fasting blood glucose,glycated hemoglobin,fructosamine,2-hour postprandial blood glucose,and total cholesterol(P<0.05).In addition,the treatment group showed significant reduction in the standard deviation of blood glucose,amplitude of postprandial blood glucose fluctuations,and 24-hour average blood glucose.Insulin resistance and visceral fat index were also significantly decreased in the treatment group(P<0.05).The decreases in MDA and LPO and the increases in SOD and PON1 indicated that the treatment group had better antioxidant capacity(P<0.001).There was no significant difference in the incidence of gastrointestinal adverse reactions,hypoglycemia,or liver damage between the two groups(P>0.05).Conclusion Degu asparagus insulin combined with semaglutide can effectively improve metabolic indicators of obese patients with type 2 diabetes and it provide an effective program for the comprehensive treatment of obese type 2 diabetes.

Objective To compare the changes in anorectal function between patients with Parkinson's disease constipation(PDC)and with functional constipation(FC),and further understand the constipation characteristics of PDC patients.Methods From 2017 to 2022,34 patients with PDC and 39 patients with FC who visited the Department of Neurology and the Department of Gastroenterology in Xuanwu Hospital,Capital Medical University were selected as the observation group and the control group.High resolution anorectal manometry was used to detect the motility,sensation and reflex of anorectal sphincter and pelvic floor muscles in the two groups of patients with constipation,and to compare the anorectal function changes of the two groups of patients.according to the results of manometry,the characteristics of constipation in the two groups of patients were analyzed and compared.Results The resting anal sphincter pressure and intrarectal pressure in the PDC group were significantly lower than those in the FC group,with statistical significance[(71.73±20.01)mmHg vs(85.02±19.74)mmHg,(22.30±21.12)mmHg vs(38.10±17.00)mmHg,respectively,1 mmHg=0.133 kPa,all P＜0.05].The length of high pressure zone,maximum squeeze pressure,continuous systolic time and anal relaxation rate in PDC group were lower than those in FC group,but no statistically significant differences;The maximum tolerance threshold of patients in the FC group was significantly higher than that in the PDC group,but there was no statistically significant difference between the two groups.There was no statistically significant difference in rectanal inhibitory reflex,initial sensory threshold,initial defecation threshold,and rectal compliance between the two groups.According to the pressure measurement results,it was found that both groups of patients mainly had insufficient defecation thrust.The proportions of insufficient defecation propulsion in PDC group and FC group were 88.2％(30/34)and 59.0％(23/39),the proportions of dyscoordination were 11.8％(4/34)and 30.8％(12/39),and the proportions of normal defecation propulsion without uncoordinated defecation were 0％and 10.3％(4/39),respectively.There was significant difference between the two groups in the type composition ratio of defecation disorder(x2=8.623,P＜0.05).Conclusions Both PDC patients and FC patients have abnormal rectal and anal motility and sensation,and the main manifestation of pressure measurement classification is insufficient defecation thrust.However,compared to FC patients,PDC patients have significantly lower anal resting pressure and rectal defecation pressure.In depth research on the changes in anorectal function and constipation characteristics of PDC patients can help improve understanding of the disease and provide relevant evidence for developing reasonable treatment plans for PDC.

Objective To compare the changes in anorectal function between patients with Parkinson's disease constipation(PDC)and with functional constipation(FC),and further understand the constipation characteristics of PDC patients.Methods From 2017 to 2022,34 patients with PDC and 39 patients with FC who visited the Department of Neurology and the Department of Gastroenterology in Xuanwu Hospital,Capital Medical University were selected as the observation group and the control group.High resolution anorectal manometry was used to detect the motility,sensation and reflex of anorectal sphincter and pelvic floor muscles in the two groups of patients with constipation,and to compare the anorectal function changes of the two groups of patients.according to the results of manometry,the characteristics of constipation in the two groups of patients were analyzed and compared.Results The resting anal sphincter pressure and intrarectal pressure in the PDC group were significantly lower than those in the FC group,with statistical significance[(71.73±20.01)mmHg vs(85.02±19.74)mmHg,(22.30±21.12)mmHg vs(38.10±17.00)mmHg,respectively,1 mmHg=0.133 kPa,all P＜0.05].The length of high pressure zone,maximum squeeze pressure,continuous systolic time and anal relaxation rate in PDC group were lower than those in FC group,but no statistically significant differences;The maximum tolerance threshold of patients in the FC group was significantly higher than that in the PDC group,but there was no statistically significant difference between the two groups.There was no statistically significant difference in rectanal inhibitory reflex,initial sensory threshold,initial defecation threshold,and rectal compliance between the two groups.According to the pressure measurement results,it was found that both groups of patients mainly had insufficient defecation thrust.The proportions of insufficient defecation propulsion in PDC group and FC group were 88.2％(30/34)and 59.0％(23/39),the proportions of dyscoordination were 11.8％(4/34)and 30.8％(12/39),and the proportions of normal defecation propulsion without uncoordinated defecation were 0％and 10.3％(4/39),respectively.There was significant difference between the two groups in the type composition ratio of defecation disorder(x2=8.623,P＜0.05).Conclusions Both PDC patients and FC patients have abnormal rectal and anal motility and sensation,and the main manifestation of pressure measurement classification is insufficient defecation thrust.However,compared to FC patients,PDC patients have significantly lower anal resting pressure and rectal defecation pressure.In depth research on the changes in anorectal function and constipation characteristics of PDC patients can help improve understanding of the disease and provide relevant evidence for developing reasonable treatment plans for PDC.

Objective:To investigate the current status of subjective well-being among stroke patients, and to explore the pathways and effects of influencing factors using structural equation model, so as to provide reference for improving subjective well-being among stroke patients.Methods:From July to November 2024, the stroke patients admitted to the First Affiliated Hospital of Bengbu Medical University, the Second Affiliated Hospital of Bengbu Medical University, Hefei First People′s Hospital were selected by convenience sampling method. A cross-sectional survey was conducted using a general demographic questionnaire, General Well-Being Scale, Cognitive Reserve Index questionnaire, Social Support Rating Scale, Stroke Symptom Cluster Scale, and FRAIL Scale, and AMOS 26.0 was used to analyse the pathways and effects of influencing factors of subjective well-being.Results:A total of 435 questionnaires were collected, 410 were valid.Among 410 cases, 266 case were males, 144 were females, with an age of (65.96 ± 12.15) years. The subjective well-being scores of stroke patients were (72.58 ± 11.66) points. Cognitive reserve and social support were positively correlated with subjective well-being ( r = 0.517, 0.554, both P<0.01), while symptom burden and frailty were negatively correlated with subjective well-being ( r = -0.687, -0.670, both P<0.01). Path analysis showed that symptom burden, frailty, cognitive reserve, and social support had a direct impact on subjective well-being (path coefficients were -0.500, -0.266, 0.148, and 0.144, respectively, all P<0.05), while cognitive reserve, social support, and symptom burden had an indirect impact on subjective well-being (path coefficients were 0.287, 0.249, and 0.108, respectively, all P<0.05). Conclusions:The subjective well-being of stroke patients is influenced by multiple factors, with symptom burden being an important factor affecting subjective well-being. Intervention strategies such as improving cognitive reserve, strengthening social support systems, and preventing frailty can improve the subjective well-being of patients.

Objective:To establish a general method for the simultaneous determination of hydroxyphenyl esters and quaternary ammonium bacteriostatic agents in eye drops.Methods:The chromatographic analysis was per-formed on an Agilent C18 column(4.6 mm ×250 mm,5 μm)with 1％triethylamine solution(pH adjusted to 5.0 with phosphoric acid)as mobile phase A and methanol as mobile phase B.Gradient elution was performed at col-umn temperature of 40 ℃.The detection wavelength was 214 nm,the flow rate was 1 mL·min-1,and the injec-tion volume was 20 μL.Results:Methylparaben,ethylparaben,propylparaben,butylparaben,benzalkonium chlo-ride and benzalkonium bromide were 0.11-559.0,0.10-513.0,0.10-258.8,0.11-270.5,1.07-537.0 and 1.03-512.8 μg·mL-1,respectively.The linear range was good(r＞0.999).The average recoveries of meth-ylparaben,benzalkonium bromide and benzalkonium chloride were 104.7％(RSD=1.3％),102.6％(RSD=1.1％)and 100.9％(RSD=1.1％),respectively.The contents of bacteriostatic agent in 100 batches of eye drops from 36 varieties of 12 enterprises were determined,and the accurate results were obtained.Conclusion:This meth-od provides a reference for the content quality control and safety evaluation of bacteriostatic agents in eye drops.

Objective To evaluate the safety of the low glucoside composites of Epimedii Folium and clarify the pharmacokinetic characteristics of its five low glucosides.Methods Four groups of KM mice were orally administrated of corn oil,1 968,2 625 and 3 500 mg·kg-1 low glucoside composites of Epimedii Folium,respectively.Then,the living conditions,toxic symptoms,and death of the mice were observed for 7 consecutive days.After the mice were dissected,the viscera/body ratio and the viscera/brain ratio were calculated.Besides,the contents of alanine aminotransferase(ALT)and aspartate transaminase(AST)in plasma were determined by ELISA,and the pathological changes of the liver were observed by HE staining.C57BL/6J mice were intravenously or orally administered of baohuoside I,baohuoside II,sagittatoside A,sagittatoside B and sagittatoside C.Then,blood samples were collected at different time points.The plasma concentrations of the five low glucosides were measured by UHPLC-MS/MS.Results When compared with the control group,no significant differences were found in the body mass,viscera/body ratio,viscera/brain ratio,contents of ALT and AST in plasma after oral administration of different doses of low glucoside composites to mice.Moreover,no pathological changes or damages were found in the liver sections.After intravenous injection,the AUC0-t values of baohuoside Ⅰ,baohuoside Ⅱ,sagittatoside A,sagittatoside B and sagittatoside C in mice were 4.82,82.54,276.64,88.77 and 178.02 min·μg·mL-1,respectively.Meanwhile,the t1/2 values were 60.42,115.27,67.63,131.61 and 129.87 min,respectively.After oral administration,the AUC0-t values of the five low glucosides were 31.64,18.59,3.48,2.41 and 2.42 min·μg·mL-1,respectively.The Cmax values were 147.23,86.76,15.58,24.34 and 26.12 ng·mL-1,respectively.The tmax values were 21.00,78.00,78.00,30.00 and 28.00 min,respectively.The bioavailability of baohuosideⅠ,baohuosideⅡ,sagittatoside A sagittatoside B and sagittatoside C were 1.91%,0.51%,0.05%,0.06%and 0.04%,respectively.Conclusion The low glucoside composites of Epimedii Folium has high safety,and no hepatotoxicity were observed at dose of 3 500 mg·kg-1.The 5 low glucosides are quickly absorbed and rapidly eliminated in mice,and all of them have low bioavailability.

Objective:To discuss the inhibitory effect of Schisandrin B on the proliferation of pancreatic cancer Pan02 cells,and to clarify the mechanism.Methods:CCK-8 method was used to detect the proliferation rates of the Pan02 cells after treated with different concentrations(0,0.78,1.56,3.12,6.25,12.50,and 25.00 mg·L-1)of Schisandrin B to select the optimal concentration and treatment time of Schisandrin B.The mouse pancreatic cancer Pan02 cells were divided into control group(0 mg·L-1 Schisandrin B),2.5 mg·L-1 Schisandrin B group,5.0 mg·L-1 Schisandrin B group,and 10.0 mg·L-1 Schisandrin B group.The morpholoy of Pan02 cells invarious groups was observed with light microscope;5-ethynyl-2'-deoxyuridine(EdU)staining assay was used to detect the positive expression rates of the Pan02 cells in various groups;flow cytometry was used to detect the percentages of the Pan02 cells at different cell cycles and the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of cell cycle and apoptosis-related proteins in the cells in various groups.Results:The CCK-8 method results showed that after treated with Schisandrin B for 48 and 72 h,compared with 0 mg·L-1 Schisandrin B,the proliferation rates of the Pan02 cells after treated with different concentrations of Schisandrin B were decreased(P<0.01),especially at 72 h.0.25,5.0,and 10.0 mg·L-1 Schisandrin B were selected to treat the Pan02 cells,and 72 h was the treatment time.In control group,the Pan02 cells had a spindle shape,with good condition,and grew closely adhered to the wall with normal organelles and cytoplasm,in 2.5 and 5.0 mg·L-1 Schisandrin B groups,the cell volume was decreased,the intercellular adhesion was disappeared,and the cell membrane was intact but more permeable;the cytoplasm shrank and vacuolar structures appeared inside the cells,with some fragmented and floating on the surface of the solution;in 10.0 mg·L-1 Schisandrin B group,the Pan02 cells exhibited notable apoptotic bodies,indicating an apoptotic state.The EdU staining results showed that compared with control group,the rates of EdU positive cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.01).The flow cytometry results showed that compared with control group,the percentages of the cells at S phase in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01),while the percentages of the cells at G2/M phase were significantly decreased(P<0.01),and the percentages of the cells at G0/G1 phase in 5.0 amd 1.0 mg·L-1 Schisandrin groups were decreased(P<0.01);compared with control group,the apoptotic rates of the cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of p27,B-cell lymphoma 2(Bcl-2)associated X protein(Bax),cleaved cysteine aspartic acid protease-3(cleaved Caspase-3),and cleaved poly adenosine diphosphate(ADP)ribose polymerase(cleaved PARP)proteins in the cells in 2.5 mg·L-1 Schisandrin B group were significantly increased(P<0.05 or P<0.01),the expression levels of cyclin A2,cyclin E2,and Bcl-2 proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.05 or P<0.01),while the expression levels of p27,Bax,cleaved Caspase-3,and cleaved PARP proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).Conclusion:Schisandrin B has an inhibitory effect on proliferation of the pancreatic cancer Pan02 cells,and its mechanism may be related to the activation of the cysteine aspartic acid protease-3(Caspase-3)pathway to induce the apoptosis and activating p27 protein to induce the arrest of cell cycle at S phase.