Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund's complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-α in rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-α levels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-α levels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.

Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund's complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-α in rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-α levels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-α levels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.

Objective:To investigate the protective effect of moxibustion on joints and its influence on the expression levels of S100 calcium binding protein A8(S100A8),S100 calcium binding protein A9(S100A9),serum amyloid A1(SAA1),and related inflammatory factors in rats with adjuvant arthritis(AA).Methods:Forty Wistar rats were randomly divided into a normal group,a model group,a moxibustion group,and a medication group,with 10 rats in each group.Except for the normal group,AA models were established in the other three groups by exposing rats to wind-cold-dampness environmental conditions combined with complete Freund's adjuvant.After successful modeling,the moxibustion group received moxibustion intervention,while the medication group was administered tripterygium glycosides tablets via oral gavage.The normal and model groups underwent similar handling and fixation without additional interventions.After 15 d of intervention,hematoxylin-eosin staining was used to assess pathological changes in the knee joint synovial membrane.Western blotting was performed to detect the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue.Enzyme-linked immunosorbent assay was used to measure the serum levels of interferon(IFN)-γ,interleukin(IL)-6,and IL-23.Results:Compared to the normal group,the model group exhibited significantly increased protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue,as well as elevated serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Histopathological analysis revealed marked synovial hyperplasia and extensive infiltration of inflammatory cells in the model group.Compared to the model group,both the moxibustion and medication groups showed significant reductions in the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue,as well as decreased serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Additionally,synovial tissue in these two groups displayed minimal hyperplasia and only mild inflammatory cell infiltration.Notably,compared to the moxibustion group,the medication group exhibited significantly higher protein expression of S100A9 in the synovial tissue(P<0.05),while no significant differences were observed in the expression of S100A8,SAA1,or serum levels of IFN-γ,IL-6,and IL-23(P>0.05).Both intervention groups showed comparable degrees of synovial inflammation,clear tissue structure,and no obvious hyperplasia.Conclusion:Moxibustion can alleviate joint swelling and reduce inflammatory responses in AA rats.Its mechanism may involve regulating the protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue.

Objective:To investigate the protective effect of moxibustion on joints and its influence on the expression levels of S100 calcium binding protein A8(S100A8),S100 calcium binding protein A9(S100A9),serum amyloid A1(SAA1),and related inflammatory factors in rats with adjuvant arthritis(AA).Methods:Forty Wistar rats were randomly divided into a normal group,a model group,a moxibustion group,and a medication group,with 10 rats in each group.Except for the normal group,AA models were established in the other three groups by exposing rats to wind-cold-dampness environmental conditions combined with complete Freund's adjuvant.After successful modeling,the moxibustion group received moxibustion intervention,while the medication group was administered tripterygium glycosides tablets via oral gavage.The normal and model groups underwent similar handling and fixation without additional interventions.After 15 d of intervention,hematoxylin-eosin staining was used to assess pathological changes in the knee joint synovial membrane.Western blotting was performed to detect the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue.Enzyme-linked immunosorbent assay was used to measure the serum levels of interferon(IFN)-γ,interleukin(IL)-6,and IL-23.Results:Compared to the normal group,the model group exhibited significantly increased protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue,as well as elevated serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Histopathological analysis revealed marked synovial hyperplasia and extensive infiltration of inflammatory cells in the model group.Compared to the model group,both the moxibustion and medication groups showed significant reductions in the protein expression of S100A8,S100A9,and SAA1 in the synovial tissue,as well as decreased serum levels of IFN-γ,IL-6,and IL-23(P<0.01).Additionally,synovial tissue in these two groups displayed minimal hyperplasia and only mild inflammatory cell infiltration.Notably,compared to the moxibustion group,the medication group exhibited significantly higher protein expression of S100A9 in the synovial tissue(P<0.05),while no significant differences were observed in the expression of S100A8,SAA1,or serum levels of IFN-γ,IL-6,and IL-23(P>0.05).Both intervention groups showed comparable degrees of synovial inflammation,clear tissue structure,and no obvious hyperplasia.Conclusion:Moxibustion can alleviate joint swelling and reduce inflammatory responses in AA rats.Its mechanism may involve regulating the protein expression of S100A8,S100A9,and SAA1 in the knee joint synovial tissue.

Objective: To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid (NMDA) receptor subtype 2B (NR2B) in the hippocampus of rheumatoid arthritis (RA) rats, and to explore the analgesic mechanisms of moxibustion in RA treatment. Methods: Sixty male Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, a moxibustion + NMDA receptor antagonist (AP-5) group, and a moxibustion + NMDA receptor agonist (NMDA) group, with 12 rats in each group. Except for the normal group, rats in the other four groups were treated with complete Freund's adjuvant in a windy, cold, and damp environment to replicate RA models. Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu (BL23) and Zusanli (ST36), and the two points were used alternately. After intraperitoneal injection of AP-5 or NMDA, rats in the moxibustion + AP-5 group and the moxibustion + NMDA group received the same moxibustion intervention as in the moxibustion group, once a day for 15 d. The thermal withdrawal latency (TWL) of rats in each group was detected before and after modeling and after the 15-day intervention. After the 15-day intervention, hematoxylin-eosin staining was performed to observe the pathological changes in knee joints. The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus; Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B. Results: The synovial tissue was proliferated, the synovial lining was significantly thickened, the pannus was formed, and the cartilage and bone tissues were significantly damaged in the model group. After intervention, the pathological morphology of the knee joints in the moxibustion group, the moxibustion + AP-5 group, and the moxibustion + NMDA group was significantly improved, and the improvement in the moxibustion + AP-5 group was more notable than that in the moxibustion + NMDA group. Compared with the normal group, the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group (P<0.01). Compared with the model group, the TWL of each intervention group was significantly increased (P<0.01 or P<0.05), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased (P<0.01). Compared with the moxibustion group, the TWL was significantly increased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion + AP-5 group (P<0.01); the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion + NMDA group (P<0.01). Conclusion: Moxibustion reduces hyperalgesia in RA inflammatory rats. The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.

Objective To evaluate the effect of ulinastatin pretreatment on endoplasmic reticulum stress during myocardial injury in the patients undergoing mitral valve replacement (MVR) with cardiopulmonary bypass (CPB).Methods One hundred patients of both sexes,aged 35-64 yr,weighing 40-80 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ (New York Heart Association Ⅱ or Ⅲ),scheduled for elective MVR with CPB,were divided into ulinastatin pretreatment group (UP group,n=50) and normal saline control group (NS group,n =50) using a random number table.Ulinastatin 0.5× 104 U/kg was intravenously infused over 1 h before skin incision,and administration was repeated every 4 h until the end of operation in group UP,while the equal volume of normal saline was given instead in group NS.Immediately after opening the right atrium (T0),at 30 min after aortic clamping (T1) and while suturing the right atrium (T2),blood samples were collected from the radial artery for measurement of the concentrations of plasma creatine kinase-MB and cardiac troponin T by enzyme-linked immunosorbent assay.Right auricle specimens were obtained after blood sampling at each time point for determination of the expression of glucose-regulated protein 78,CCAAT/enhancer-binding protein homologous protein and c-Jun N-terminal kinase protein and mRNA (by real-time polymerase chain reaction and Western blot,respectively) and apoptosis in cardiomyocytes (by TUNEL).The apoptosis rate was calculated.Results Compared with group NS,the plasma concentrations of creatine kinase-MB and cardiac troponin T at T1 and T2 and apoptosis rate at T2 were significantly decreased,and the expression of glucose-regulated protein 78,CCAAT/enhancer-binding protein homologous protein and c-Jun N-terminal kinase protein and mRNA was down-regulated at T1 and T2 in group UP (P＜0.05).Conclusion The mechanism by which ulinastatin pretreatment inhibits apoptosis in cardiomyocytes and attenuates myocardial injury is related to decrease in endoplasmic reticulum stress in the patients undergoing MVR with CPB.

Objective To summarize the preliminary experience and early -stage follow -up results of simple transesophageal echocardiography(TEE)to guide right jugular internal vein for closure of atrial septal defect (ASD), and to investigate its feasibility.Methods A total of 24 ASD patients(12 male,12 female)treated by transcatheter ASD closure under solely guidance of TEE were summarized in Children Heart Center,People′s Hospital of Zhengzhou University from February 2015 to February 2016.Mean age,body weight and average diameter of the 24 cases of ASD were 5 -18 (12.4 ±5.6)years old,16 -60 (34.1 ±9.6)kg,5 -20 (11.0 ±2.5)mm,respectively.All patients were treated by right jugular internal vein transcatheter closure under solely guidance of TEE.The efficiency of imme-diate post -operation under TEE was estimated,and follow -ups were done at 24 hours,1 month,3,6 and 12 months after the procedures by transthoracic echocardiography,X -ray examination and electrocardiogram.Results A total of 23 patients were treated successfully under solely guidance of TEE.The diameter of devices was (14.5 ±3.6)mm.The procedural time was(20.5 ±7.2)min.One patient who had 2 ASD was treated by traditional thoracic surgery,because the small ASD had 5 mm residual shunt after the procedures.There were 3 cases with trivial residual shunt after the pro-cedures immediately.All patients could move at once after operation.The average follow -up time was(5.6 ±1.2) months.No patients suffered from residual shunt,occluder malposition,peripheral vascular injury,pericardial effusion or cardiac perforation during the follow -up.Conclusions For some selected patients,simple TEE guiding right jugular in-ternal vein for closure of ASD is applicable.The method not only prevents injury from radiation,but also shortens the in -bed time and avoid the lower limbs braking,with good early -stage follow -up results are desirable.

Objective: To explore the blood-saving effect of autologous platelet-rich plasma (PRP) back-transfusion in patients with Stanford type A aortic dissection surgery. Methods: A total of 59 consecutive patients who received Stanford type A aortic dissection surgery in our hospital from 2013-01 to 2015-10 were studied. The patients were at the age of (50±6) years with mean body weighting at (80±12) kg and were randomly divided into 2 groups: Traditional (T) group,n=31 and Autologous PRP back-transfusion (P) group,n=28. Blood levels of Hb, platelet counts, PT, APTT were measured at pre-induction of anesthesia (T1), before CPB (T2), prior ifnishing of CPB (T3) and at 1 h (T4), 24 h (T5), 48 h (T6) after the operation. The in-operative, 48 h post-operative volumes of allogeneic blood transfusion and the volume of chest tube drainage at 48h after operation were recorded; the complication occurrence at peri-operative period was recorded. Results: In P group, whole blood processing volume was (1269±197) ml, PRP volume was (753±78) ml, PRP separation time was (35±9) min and the separated platelets were about (22±3)% of total platelet counts. Compared with T group, P group had decreased Hb at T2 (131.0±15.0) g/L vs (101.0±10.0) g/L, decreased platelet counts at T3 (115.0±51.0)×109 /L vs (83.0±23.0)×109/L, while increased platelet counts at T4 (103.0±25.0)×109/L vs (151.0±27.0)×109/L, T5 (105.0±25.0)×109 /L vs (147.0±39.0)×109/L and T6 (101.0±26.0) ×109/L vs (149.0±35.0)×109/L, allP<0.05; P group presented reduced PT at T4 (17.6±2.1) s vs (14.1±1.1) s and T5 (17.6±2.7) s vs (13.5±0.8) s, allP<0.05. The in-operative transfusions of platelet, plasma, cold precipitation and post-operative transfusions of red blood cells, platelets, plasma, cold precipitation and the volume of chest tube drainage at 48h after operation were less in P group,P<0.05. Compared with T group, P group had the lower rates of acute post-operative lung injury (32.1% vs 19.4%), shorter mechanical ventilation time (69.1±5.9) h vs (43.1±1.5) h and ICU staying time (8.1±2.8) d vs (5.3±1.1) d, allP<0.05. Conclusion: Autologous PRP back-transfusion could reduce the post-operative bleeding and allogeneic blood transfusion for Stanford A aortic dissection surgery, it has obvious blood-saving effect.

ObjectiveTo study the effect of anticoagulant therapy with low molecular weight heparin (LMWH) on coagulation and inflammation markers in sepsis patients.Methods A prospective randomized controlled trial was conducted. Sixty sepsis patients admitted to intensive care unit (ICU) of Zhengzhou University People's Hospital from March 2012 to May 2014 were divided into control group and observation group, with 30 cases in each group. The observations were begun as soon as the diagnosis of sepsis was established, and the observation time was 7 days. All sepsis patients were treated according to the 2008 international sepsis treatment guidelines. Every patient in the observation group was subcutaneously injected with LMWH 0.6 mL on the first day of ICU admission, twice a day for 7 days. The blood from peripheral vein was collected at 1, 3, 5, 7 days of treatment, and CD62p, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), D-dimmer was determined by immunoturbidimetry, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score was recorded, and incidence of multiple organ dysfunction syndrome (MODS) was also evaluated.Results There were no significant differences in values of all parameters, including CD62p, D-dimmer, IL-6, TNF-α, and APACHEⅡ score at 1 day of treatment. The values of all parameters in observation group were gradually decreased. CD62p at 3 days of treatment and D-dimmer, IL-6, TNF-α, and APACHEⅡ score at 5 days of treatment were significantly lower than those at 1 day of treatment. The values in the control group were decreased at first and then increased, as D-dimmer, IL-6 and TNF-α were significantly higher on the 5th day than those at 1 day of treatment. Compared with control group, CD62p, D-dimmer, IL-6, TNF-α and APACHEⅡ score on the 7th day of treatment were significantly lowered in observation group [CD62 (μg/L): 22.64±2.88 vs. 31.52±2.81, D-dimmer (g/L): 1.32±0.46 vs. 4.79±0.82, IL-6 (ng/L): 5.84±1.87 vs. 49.64±3.12, TNF-α (ng/L): 21.04±3.15 vs. 130.58±6.26, APACHEⅡ score: 9.71±2.02 vs. 14.17±2.38, allP< 0.05]. Correlation analysis showed that in observation group, CD62p, D-dimmer, IL-6, and TNF-α were positively correlated with APACHEⅡ score (r value was 0.907, 0.868, 0.880, 0.693, respectively, all P=0.000). The incidence of MODS in observation group was significantly lower than that in the control group [26.7% (8/30) vs. 46.7% (14/30),χ2=3.943,P= 0.028].Conclusions LMWH, which was given early in sepsis, can significantly down-regulate the expression of CD62p, D-dimmer, IL-6 and TNF-α, and reduce the incidence of MODS. Some indicators regarding coagulation and inflammation can be used as supplementary indicators to severity scores, and it may be able to improve the accuracy of scoring systems for sepsis.