Development and verification of a rapid detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line / 中国生物制品学杂志

@#Objective To develop and verify a rapid detection method for the biological activity of adalimumab based on U937-NF-κB-Luc cell line. Methods Using U937-NF-κB-Luc cell line as the detection cells,a method for detecting the biological activity of adalimumab was developed based on luciferase luminescence principle. The method was optimized for the concentration of tumor necrosis factor-α(TNF-α)(160 ng/mL as initial concentration,2 times serial dilution,10dilutions),the initial concentration of antibody(2 000 ng/mL,2 times serial dilution,20 dilutions),the dilution multiple of antibody(1. 5,2,3,4 times),the inoculation amount(8 × 103,2 × 104,4 × 104,6 × 104cells/well)and the incubation time(0. 5,1,2,3 h),and verified for the specificity,accuracy,precision and linear range. The relative potency of five batches of adalimumab was detected by using the optimized method and TNF-α neutralization activity method based on L929cells respectively. Results The dose-response curve of adalimumab international standard showed a typical S-type,and the data complied with the four-parameter equation y =(A-D)/[1 +(x/C)B]+ D,R2> 0. 99. The optimum concentration of TNF-α was 5 ng/mL,the initial concentration of antibody was 800 ng/mL,the dilution ratio for adalimumab was 1∶2,the inoculation amount was 2 × 104cells/well,and the induction time was 2 h. Three therapeutic monoclonal antibodies of TNF-α target,such as adalimumab,obtained good dose-response curves,while therapeutic monoclonal antibodies of other non-TNF-α targets did not show this curve. The linear regression equation of the logarithmic value of theoretical potency and the logarithmic value of the corresponding measured potency had a slope of 1. 037,and the relative bias was within the range of ± 12%. The geometric coefficient of variation(GCV)of the relative titer measured value of each sample was less than20%. The theoretical potency ranged from 64% to 156%,showing a good linear relationship with the measured values,and the fitting linear regression equation was y = 1. 037 4 x-0. 023 7,R2= 0. 998 4. There was no significant difference in the relative potency measured results of five batches of adalimumab by the two methods(t = 1. 198,P = 0. 265 1). Conclusion The developed detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line has good specificity,accuracy and precision with short time consumption(3 h),which can be used as a rapid evaluation method for the biological activity of adalimumab.

Analysis of cytokines level in patients with Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis / 白血病·淋巴瘤

Objective:To investigate the level change of cytokines in patients with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (EBV-HLH).Methods:A retrospective case control study was conducted. The clinical data of 65 patients with EBV-HLH, 30 patients with infectious mononucleosis (IM) (IM group) and 40 patients with non-EBV infection-associated hemophagocytic lymphohistiocytosis (non-EBV-HLH group) who admitted to Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology from February 2022 to February 2023 were retrospectively analyzed. The enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of the interleukin (IL)-6, IL-2, IL-10, IL-8, IL-1β, tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in serum samples of patients in the above 3 groups. The cytokines levels in EBV-HLH group were compared with those in IM group and non-EBV-HLH group, respectively.Results:The cytokines levels of IL-6, IL-2, IL-10, IL-8, IL-1β, TNF-α and IFN-γ in EBV-HLH group were higher than those in the non-EBV-HLH group, and the differences were statistically significant (all P < 0.05). The cytokines levels of IL-2, IL-10 and IFN-γ in EBV-HLH group were higher than those in IM group, and the differences were statistically significant (all P < 0.05). Conclusions:The cytokines levels of IL-6, IL-2, IL-10, IL-8, IL-1β, TNF-α, IFN-γ are increased in EBV-HLH patients, which may play an important role in the development and progression of EBV-HLH.

Analysis of clinical characteristics of 33 cases of tuberculosis complicated by tumor necrosis factor-α inhibi-tor in autoimmune diseases / 实用医学杂志

Objective To investigate the clinical characteristics,treatment and prognosis of tuberculosis in patients with autoimmune diseases after tumor necrosis factor-αinhibitors.Methods Clinical data of 33 patients with TB after biologics(tumor necrosis factor-α inhibitors)treated in Guangzhou Chest Hospital from January 2019 to March 2023 were collected,including 25 males and 8 females,with a median age of 32 years.The clinical symptoms,laboratory results,imaging and tracheoscopic features,pathological features,treatment and outcome were analyzed retrospectively.Results The common clinical manifestations were cough(26/33),sputum(23/33)and fever(17/33).The most common cases were pulmonary tuberculosis(32/33),bronchial tuberculosis(15/33),mediastinum and hilar lymph node tuberculosis(11/33).Bilateral lung spread of tuberculosis(21/33),intrapulmonary spread of tuberculosis(bronchus,mediastinal hilar lymph nodes,pleura)(19/33),extrapulmonary tuberculosis(18/33),pulmonary tuberculosis with intrapulmonary or extrapulmonary tuberculosis(26/33).Blood CD4+T lymphocyte test was normal(23/33),and blood IGRA test was positive(27/33).Pulmonary imaging miliary nodules(8/33).The histopathology of the lymph nodes showed atypical granulomatous nodules.The duration of anti-tuberculosis treatment is 8-32 months.1 case of death.Conclusion Patients with autoimmune diseases complicated with tuberculosis after the application of tumor necrosis fact-α inhibitor are more likely to have double lung lesions,which are easy to spread to lung tissues and multiple organs of the body,and have decreased immune function.Most of them need to extend the treatment course,and the prognosis is generally good after comprehensive treatment.

Mechanism by which miR-34a-5p/PLCD3 axis regulates osteoarthritis progression / 中国组织工程研究

BACKGROUND:Molecular mechanisms targeting the miRNA/mRNA axis to regulate osteoarthritis disease process have been studied.We identified the mRNA:phospholipase C delta 3(PLCD3)and its target miRNA(miR-34a-5p)with clinical predictive value through previous bioinformatics studies,while experiments to verify their specific roles and mechanisms in regulating osteoarthritis are still lacking. OBJECTIVE:To investigate the regulatory role and mechanism of miR-34a-5p/PLCD3 axis on osteoarthritis progression. METHODS:The synovium of 15 patients with knee osteoarthritis was selected as the osteoarthritis group,and the synovium of 15 young patients with internal fixation of patellar fracture caused by trauma during the same period was selected as the control group.The expression of PLCD3 and miR-34a-5p in the synovium was detected by real-time PCR.Human fibroblast like synovial cells-osteoarthritis(HFLS-OA)cells were treated by cell transfection and divided into miR-34a-5p mimic group,pCDH-PLCD3 group,miR-34a-5p mimic+pCDH-PLCD3 group,miR-34a-5p inhibitor group,si-PLCD3 group,and miR-34a-5p inhibitor+si-PLCD3 group.The relationship between PLCD3 and miR-34a-5p expression was detected by real-time PCR.The effects of HFLS-OA cell viability and cell migration in each group were detected by CCK-8 assay and cell scratch test.Western blot assay was used to detect the expression level of apoptosis marker protein.The expression of inflammatory factors was detected by ELISA. RESULTS AND CONCLUSION:(1)PLCD3 was a direct target of miR-34a-5p,and the expression levels of PLCD3 and miR-34a-5p were negatively correlated.(2)Upregulation of PLCD3 promoted proliferation of HFLS-OA cells and inhibited cell migration.The up-regulation of miR-34a-5p significantly inhibited the activity of HFLS-OA cells and enhanced cell migration.Overexpression of miR-34a-5p significantly increased the levels of Casp3 and Casp9 proteins in HFLS-OA cells,while overexpression of PLCD3 showed the opposite trend.(3)PLCD3 overexpression significantly increased the expression of interleukin 6 and tumor necrosis factor alpha in HFLS-OA cells,while miR-34a-5p mimics showed protective activity.(4)The miR-34a-5p/PLCD3 axis may affect the progression of osteoarthritis by regulating the inflammatory process or apoptosis of synovial cells.

Efficacy of rituximab injection combined with CHOP regimen in the treatment of diffuse large B-cell lymphoma / 中国医师进修杂志

Objective:To evaluate the clinical efficacy of rituximab injection combined with CHOP regimen (cyclophosphamide+doxorubicin+vincristine+prednisolone) in the treatment of diffuse large B-cell lymphoma (DLBCL).Methods:One hundred and twenty patients with DLBCL who treatment in the First Affiliated Hospital of Xinjiang Medical University from June 2019 to June 2022 were selected as the study object. They were randomly divided into the study group (60 cases) and the control group (60 cases). The control group was treated with CHOP regimen, and the study group was treated with rituximab injection on the basis of CHOP regimen. The clinical efficacy, inflammatory reaction, immune function and adverse reaction were evaluated after 6 courses of treatment.Results:After treatment, the total clinical effective rate in the study group was higher than that in the control group: 88.33%(53/60) vs. 70.00%(42/60), there was statistical difference ( χ2 = 6.11, P<0.05). Before treatment, the levels of serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the two groups had no significant differences ( P>0.05); after treatment, the levels of serum IL-6 and TNF-α were decreased, and the levels of serum IL-6 and TNF-α in the study group were lower than those in the control group: (223.56 ± 21.28) ng/L vs. (267.35 ± 25.36) ng/L, (9.34 ± 2.75) μg/L vs. (11.96 ± 3.83) μg/L, there were statistical differences ( P<0.05). Before treatment, the levels of serum immunoglobulin (Ig) A, IgM and IgG in the two groups had no significant differences ( P>0.05); after treatment, the levels of serum IgA, IgM and IgG were decreased, but the levels of serum IgA, IgM and IgG in the study group were higher than those in the control group: (1.83 ± 0.46) g/L vs. (1.34 ± 0.34) g/L, (1.15 ± 0.22) g/L vs. (0.83 ± 0.24) g/L, (10.67 ± 1.65) g/L vs. (8.02 ± 1.62) g/L, there were statistical differences ( P<0.05). After treatment, the incidence of thrombocytopenia, leucopenia, gastrointestinal reaction, bone marrow suppression and liver function injury in the study group were lower than those in the control group: 6.67%(4/60) vs. 20.00%(12/60), 15.00%(9/60) vs. 31.67%(19/60), 30.00%(18/60) vs. 58.33%(35/60), 5.00%(3/60) vs. 16.67%(10/60), 10.00%(6/60) vs. 25.00%(15/60), there were statistical differences ( χ2 = 4.62, 4.66, 9.77, 4.33, 4.88, P<0.05). Conclusions:The treatment effect of rituximab injection combined with CHOP regimen in DLBCL is significant, which can reduce the inflammatory reaction of the body, reduce the damage of immune function, and reduce the adverse reactions of chemotherapy.

Therapeutic efficacy of automatic peritoneal dialysis on elderly patients with cardiorenal syndrome / 中华老年心脑血管病杂志

Objective To explore the therapeutic efficacy of automatic peritoneal dialysis on elderly patients with cardiorenal syndrome(CRS).Methods A total of 260 elderly CRS patients treated at our hospital from January 2019 to January 2022 were recruited,and then randomly divided into an observation group and a control group,with 130 cases in each group.The control group received conventional basic treatment and symptomatic treatment,while the observation group received automated peritoneal dialysis treatment on this basis.Cardiac function indicators,renal function indicators,inflammatory factors,MAP and heart rate were compared between the two groups.Re-sults After treatment,significantly lower LVESD(26.29±1.19 mm vs 29.59±1.84 mm),LVEDD(47.43±1.39 mm vs 51.81±1.34 mm),LAD(30.74±1.15 mm vs 33.11±0.88 mm),and levels of NT-proBNP(1034.74±313.61 ng/L vs 2634.02±853.67 ng/L),urea(16.69±3.57 mmol/L vs 32.67±4.54 mmol/L),cystatin C(0.47±0.13 mg/L vs 0.61±0.15 mg/L),creatinine(254.74±41.15 mmol/L vs 394.09±38.61 mmol/L),TNF-α(144.14±23.16 mg/L vs 183.97± 23.37 mg/L)and hs-CRP(4.09±1.03 μg/L vs 5.45±1.17 μg/L),and higher LVEF(39.14± 4.48％vs 35.64±5.27％)were observed in the observation group than the control group(all P＜0.01).There were no significant differences in heart rate and MAP between the two groups before and after treatment(P＞0.05).Conclusion Automatic peritoneal dialysis can improve the cardiac and renal function and reduce inflammatory response in elderly CRS patients,and show positive significance for improving prognosis.

Effect and mechanism of arctigenin on ventricular remodeling and inflammatory reaction in rats with chronic heart failure / 中华老年心脑血管病杂志

Objective To investigate the influences of arctigenin(ATG)on ventricular remodeling and inflammatory reaction in chronic heart failure(CHF)rats,and to analyze its potential mecha-nism.Methods A total of 79 SD rats were randomly divided into sham operation group(n=12),and the remaining rats were inflicted with abdominal aortic coarctation to establish a rat CHF model.After modeling,60 CHF rats were randomly divided into CHF group,low and high dose ATG group(ATG-L and ATG-H groups,10 and 20 mg/kg,respectively),ATG+NC group[20 mg/kg ATG+100 μl high mobility group protein B1(HMGB1)negative control plasmid],and ATG+HMGB1 group(20 mg/kg ATG+100 pl HMGB1 overexpression plasmid),with 12 rats per group.After 4 weeks of corresponding intervention,heart function,levels of B-type brain na-triuretic peptide(BNP),N-terminal B-type brain natriuretic peptide precursor(NT-proBNP)andIL-6 and TNF-α,heart mass index(HMI)and left ventricular mass index(LVMI),pathological changes of myocardial tissue,cross-sectional area of myocardial cells and myocardial collagen vol-ume fraction(CVF)and protein expression of HMGB1/Toll-like receptor 4(TLR4)/NF-κB sig-naling pathway in left ventricular myocardial tissue were measured.Results Compared with the sham operation group,myocardial tissue HMGB1(0.42±0.05 vs 0.15±0.02)and TLR4(0.70± 0.09 vs 0.21±0.04)protein levels,and phosphorylated NF-κB p65(p-NF-κB p65)/NF-κB p65(0.73±0.09 vs 0.26±0.05)protein ratio were obviously increased in the CHF group,while the left ventricular ejection fraction(LVEF)and left ventricular short-axis fractional shortening(LVFS)were obviously decreased(P＜0.05).Myocardial tissue HMGB1(0.33±0.04、0.24±0.04 vs 0.42±0.05)and TLR4(0.56±0.06、0.41±0.05 vs 0.70±0.09)protein levels,and p-NF κB p65/NF-KB p65(0.61±0.08、0.49±0.06 vs 0.73±0.09)protein ratio were decreased,and the LVEF and LVFS were increased in the ATG-L group and ATG-H group than the CHF group(P＜0.05).Overexpression of HMGB1 obviously attenuated the inhibitory effects of ATG on HMGB1/TLR4/NF-κB signaling pathway,ventricular remodeling,and inflammatory reaction in CHF rats(P＜0.05).Conclusion ATG may suppress ventricular remodeling in CHF rats by in-hibiting HMGB1/TLR4/NF-κB signaling inflammatory pathway.

Effects of electroacupuncture on gut microbiota and related inflammatory factors in rats with Crohn disease / 针灸推拿医学（英文版）

Objective:To observe the effects of electroacupuncture(EA)on gut microbiota and serum inflammatory factors interleukin(IL)-1β and tumor necrosis factor(TNF)-α in Crohn disease(CD)model rats. Methods:Thirty-six Sprague-Dawley rats were randomly divided into a normal control(NC)group with 10 rats and a modeling group with 26 rats.In the modeling group,the CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.After successful modeling,the rats were randomly divided into a CD model(CD)group,an EA group,and a Western medicine(WM)group.The NC and CD groups received no treatment;the EA group was treated with EA for 20 min each time,with 7 consecutive days'intervention;the WM group received mesalazine enteric-coated tablet solution by gavage once a day for 7 d.The changes in body mass and disease activity index(DAI)were observed.Serum IL-1β and TNF-α were determined by enzyme-linked immunosorbent assay.Hematoxylin-eosin staining was used to observe the pathological changes of colon tissues,and 16S rDNA sequencing was used to analyze the structural changes of gut microbiota. Results:Compared with the NC group,the body mass of rats in the CD group decreased(P<0.01),and the DAI score increased(P<0.01);the colon tissue structure was disordered,and many inflammatory cells were present;also,IL-1β and TNF-α increased significantly(P<0.01).As a result,the diversity of gut microbiota decreased,and the abundance of some conditional pathogenic bacteria(such as Prevotella)increased,while the abundance of beneficial bacteria(such as Lactobacillus,Rochella,and Spirillum)decreased.After the intervention,compared with the CD group,the body mass of rats in the EA group and WM group increased(P<0.01);the DAI score decreased(P<0.01),the colon tissue structure improved,and the IL-1β and TNF-α levels decreased(P<0.01);the diversity of gut microbiota increased(P<0.05),and the abundance of some conditional pathogenic bacteria decreased while the abundance of beneficial bacteria increased in the EA group;whereas the diversity of gut microbiota in the WM group was not statistically different(P>0.05). Conclusion:EA can reduce the damage of colon mucosa,regulate the imbalance of gut microbiota,and inhibit the serum inflammatory factor IL-1β and TNF-α expression in CD rats.

Mechanism of Modified Erchentang on COPD Inflammation Based on TNF-α/TNFR1/RIPKs Pathway / 中国实验方剂学杂志

ObjectiveBased on tumor necrosis factor alpha （TNF-α）/tumor necrosis factor receptor 1 （TNFR1）/receptor-interacting protein kinases （RIPKs） signaling pathway， this paper aims to study the effect of modified Erchentang on inflammation in rats with chronic obstructive pulmonary disease （COPD） and explore its mechanism of action. MethodA total of 60 SD rats were randomly divided into normal group， model group， high， medium， and low-dose groups （20， 10， 5 g·kg-1·d-1） of modified Erchentang， and Xiaokechuan group （3.5 mL·kg-1·d-1）， with 10 rats in each group. The COPD rat model was established by cigarette smoke combined with lipopolysaccharide （LPS）. The normal group and model group were given the same amount of normal saline for 21 days by gavage administration. The contents of TNF-α and TNFR1 in bronchoalveolar lavage fluid （BALF） of rats were detected by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA expressions of RIPK1， RIPK3， and mixed lineage kinase domain-like （MLKL） in the lung tissue. The protein expressions of RIPK1， RIPK3， and MLKL in the lung tissue were detected by Western blot. The pathological changes in lung tissue were observed by hematoxylin-eosin （HE） staining. ResultCompared with the normal group， the contents of TNF-α and TNFR1 in BALF of the model group were significantly increased （P<0.01）， and the mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were significantly increased （P<0.01）. Compared with the model group， the contents of TNF-α and TNFR1 in BALF of high， medium， and low-dose groups of modified Erchentang and Xiaokechuan group were decreased （P<0.01）. The mRNA and protein expression levels of RIPK1， RIPK3， and MLKL in the lung tissue were decreased to different degrees （P<0.05， P<0.01）. ConclusionModified Erchentang can effectively improve the inflammatory response of lung tissue in COPD rats， and the mechanism may be by inhibiting the activation of the TNF-α/TNFR1/RIPKs signaling pathway.

Up-regulation of Proinflammatory Cytokines in Rostral Ventromedial Medulla Contributes to Chronic Postsurgical Pain by Promoting 5-HT Release / 中山大学学报(医学科学版)

ObjectiveTo investigate the role of proinflammatory cytokines tumor necrosis factor alpha （TNFα） and interleukin-1β （IL-1β） in rostral ventromedial medulla （RVM） in chronic postsurgical pain （CPSP） induced by skin/muscle incision and retraction （SMIR）. MethodsSD rats were randomly divided into 5 groups： ① Sham group； ② SMIR group； ③ SMIR+TNFα/IL-1β neutralizing antibody group； ④ SMIR+TNFα/IL-1β group and ⑤ SMIR+vehicle group. 50% paw mechanical withdrawal threshold （MWT） was measured by the up-down method， immunofluroscence was used to detect the TNFα and IL-1β expression and ELISA for the 5-Hydroxytryptamine （5-HT） level. ResultsSMIR elicited persistent nociceptive sensitization， upregulated TNFα and IL-1β expression in RVM neurons and astrocytes. Microinjection of TNFα or IL-1β neutralizing antibody into RVM inhibited the development of nociceptive sensitization and decreased the level of 5-HT in both RVM and spinal dorsal horn. While microinjection of recombinant TNFα or IL-1β into RVM enhanced the development of nociceptive sensitization and increased the level of 5-HT in both RVM and spinal dorsal horn. ConclusionUp-regulation of proinflammatory cytokines in RVM may contribute to SMIR induced CPSP by promoting 5-HT release.

Ginkgo ketone ester dropping pills combined with agatroban injection for the treatment of acute cerebral infarction / 中国基层医药

Objective:To investigate the clinical efficacy of ginkgo ketone ester dropping pills combined with agatroban injection in the treatment of acute cerebral infarction.Methods:This prospective case-control study was conducted on 120 patients with acute cerebral infarction who were treated at The Hospital of Shanxi University of Chinese Medicine between April 2020 and April 2022. These patients were randomly divided into a control group and a study group using the random number table method, with 60 patients in each group. The control group received intravenous injections of agatroban based on conventional treatment, while the study group received treatment with ginkgo ketone ester dropping pills combined with agatroban injection based on conventional treatment. The treatment duration was 2 weeks. Clinical efficacy was evaluated after continuous treatment for 2 weeks.Results:The overall response rate in the study group was 95.0% (57/60), which was significantly higher than 80.0% (48/60) in the control group ( χ2 = 6.17, P = 0.012). After treatment, the Barthel index in the study group was (65.3 ± 7.3) points, which was significantly higher than (59.8 ± 7.5) points in the control group ( t = -4.07, P < 0.001). The modified Rankin Scale score and the National Institutes of Health Stroke Scale score in the study group were (1.2 ± 0.4) points and (4.6 ± 0.7) points, which were significantly lower than (2.4 ± 0.6) points and (7.6 ± 1.1) points, respectively, in the control group ( t = 12.89, 17.82, both P < 0.001). Interleukin-6, hypersensitive C-reactive protein, and tumor necrosis factor-α levels in the study group were significantly lower than those in the control group ( t = 10.10, 18.25, 14.15, all P < 0.001). The nitric oxide levels in the study group were significantly higher than those in the control group, while endothelin 1 and thromboxane A2 levels in the study group were significantly lower than those in the control group ( t = -7.65, 10.77, 21.90, all P < 0.001). There was no significant difference in incidence of adverse reactions between the two groups ( P > 0.05). Conclusion:The combination of ginkgo ketone ester dropping pills and agatroban injection has a remarkable therapeutic effect on acute cerebral infarction. The combined therapy can reduce the severity of neurological deficits in patients, promote brain function recovery, improve quality of life, adjust serum inflammatory factors, and thereby be worthy of clinical application.

Clinical efficacy of Danggui Shaoyao powder combined with folic acid tablets in the treatment of chronic atrophic gastritis / 中国基层医药

Objective:To investigate the clinical efficacy of Danggui Shaoyao powder combined with folic acid tablets in the treatment of chronic atrophic gastritis. Methods:A total of 90 patients with chronic atrophic gastritis who were treated at Longyou Branch, Sir Run Run Shaw Hospital, Zhejiang University from March 2021 to March 2022 were included in this study. They were randomly divided into a control group and an experimental group, with 45 patients per group. The control group was treated with folic acid tablets, while the experimental group was treated with Danggui Shaoyao powder combined with folic acid tablets. Before and after treatment, traditional Chinese medicine syndrome scores, gastroscopy and pathology scores, and inflammatory factor levels were compared between the two groups. Clinical efficacy was compared between the two groups. Results:The amplitudes of decreases in traditional Chinese medicine syndrome score in the experimental group [acid reflux (0.57 ± 0.19) points and epigastric pain (0.84 ± 1.36) points] were significantly greater than those in the control group [acid reflux (1.46 ± 0.39) points, epigastric pain (1.52 ± 1.41) points, t = 4.86, 6.52, both P < 0.05]. Inflammatory factor levels in the experimental group [tumor necrosis factor-α (1.03 ± 0.11) μg/L, interleukin-6 (8.15 ± 1.42) ng/L, C-reactive protein (6.55 ± 0.98) mg/L] were significantly lower than those in the control group [tumor necrosis factor-α (1.73 ± 0.14) μg/L, interleukin-6 (12.24 ± 1.48) ng/L, C-reactive protein (10.23 ± 1.03) mg/L, t = 4.52, 7.66, 6.95, all P < 0.05]. The gastroscopy and pathological scores in the experimental group [intestinal metaplasia (0.78 ± 0.35) points, dysplasia (0.30 ± 0.10) points] were significantly lower than those in the control group [intestinal metaplasia (1.31 ± 0.38) points, dysplasia (0.68 ± 0.12) points, t = 4.13, 3.85, both P < 0.05]. The overall response rate in the experimental group was 93.33% (42/45), which was significantly higher than 77.78% (35/45) in the control group ( χ2 = 4.40, P < 0.05). Conclusion:Danggui Shaoyao powder combined with folic acid tablets in the treatment of chronic atrophic gastritis can effectively improve traditional Chinese medicine syndromes, reduce the level of inflammatory factors, and have a good clinical effect.

Effect of recombinant human epidermal growth factor combined with sodium hyaluronate eye drops in patients with cataract after multifocal intraocular lens implantation / 中国基层医药

Objective:To investigate the clinical efficacy of recombinant human epidermal growth factor combined with sodium hyaluronate eye drops in the treatment of cataracts after multifocal intraocular lens implantation and its effect on inflammation factors in tears and tear film stability.Methods:A total of 86 patients with cataracts who underwent multifocal intraocular lens implantation at Jinan 2 nd People's Hospital from July 2020 to January 2023 were included in this randomized controlled study. These patients were randomly divided into a control group and a combined group, with 43 patients in each group. Patients in the control group were administered sodium hyaluronate eye drops postoperatively, while patients in the combined group received a combination of recombinant human epidermal growth factor and sodium hyaluronate eye drops. All patients were treated for 1 month. Before and after treatment, the levels of inflammatory factors in tears, tear film stability-related indicators, and corneal endothelial cells were measured and compared between the two groups. Additionally, any adverse reactions experienced by the patients were recorded throughout the treatment period. Results:After treatment, the levels of interleukin-6 and tumor necrosis factor-α in the tear fluid of the combined group were (17.91 ± 2.45) μg/L and (72.14 ± 8.43) μg/L, respectively. These values were significantly lower than those in the control group, which were (24.63 ± 3.05) μg/L and (86.97 ± 9.85) μg/L, respectively ( t = 11.26, 7.50, both P < 0.001). Additionally, the fluorescein staining score for corneal damage in the combined group was (2.34 ± 0.37) points. This was significantly lower than the score of (3.42 ± 0.48) points observed in the control group ( t = 11.69, P < 0.001). Tear break-up time and Schirmer I Test in the combined group were (8.68 ± 0.96) seconds and (9.31 ± 1.04) mm/5 minutes, respectively. These values were significantly higher than those in the control group, which were (7.81 ± 0.89) seconds and (7.14 ± 0.86) mm/5 minutes, respectively ( t = -4.36, -10.54, both P < 0.001). Furthermore, the corneal endothelial cell density and the proportion of hexagonal cells in the combined group were (2 514.09 ± 259.31) counts/mm 2 and (41.67 ± 5.05)%, respectively. These values were significantly higher than those in the control group, which were (2 244.82 ± 253.37) counts/mm 2 and (36.75 ± 4.96)% in the control group ( t = -4.87, -29.45, both P < 0.001). The incidence of adverse reactions in the combined group was 11.63% (5/43), which was significantly higher than 6.98% (3/43) in the control group ( χ2 = 0.55, P > 0.05). Conclusion:The combination of recombinant human epidermal growth factor with sodium hyaluronate eye drops following multifocal intraocular lens implantation in patients with cataracts effectively decreases the levels of inflammatory factors in tear fluid. This treatment regimen also enhances tear film stability, promotes the repair of injured corneal tissue, and is highly safe.

Effect of amlodipine/benazepril tablets on blood pressure and cardiac function in older adults with hypertension complicated by coronary heart disease / 中国基层医药

Objective:To investigate the effect of amlodipine/benazepril tablets on blood pressure control and cardiac function improvement in older adult patients with hypertension complicated by coronary heart disease.Methods:A randomized controlled clinical study was conducted on 98 older adult patients with hypertension complicated by coronary heart disease who were diagnosed and treated at Liaoning Jinqiu Hospital between February 2020 and February 2022. The patients were randomly divided into an observation group and a control group, with 49 patients in each group using a random lottery method. The control group was treated with amlodipine tablets, while the observation group was treated with amlodipine/benazepril tablets for 3 months. Blood pressure levels and cardiac function improvement were compared between the two groups.Results:Prior to treatment, there were no statistically significant differences in diastolic blood pressure, systolic blood pressure, cardiac function indices, and inflammatory factor levels between the two groups (all P > 0.05). After treatment, the observation group exhibited lower levels of diastolic blood pressure and systolic blood pressure [(88.74 ± 4.26) mmHg, (125.47 ± 6.23) mmHg, 1 mmHg = 0.133 kPa] compared with the control group [(95.71 ± 4.55) mmHg, (134.28 ± 6.10) mmHg, t = 7.07, 7.82, both P < 0.001]. After treatment, the observation group showed lower levels of endothelin 1 and higher levels of nitric oxide compared with the control group ( t = 5.02, 4.96, both P < 0.05). After treatment, the observation group demonstrated lower left ventricular end-systolic diameters [(44.04 ± 3.26) mm vs. (48.58 ± 3.19) mm, t = 6.96, P < 0.001], lower left ventricular end-diastolic diameters [(52.07 ± 4.11) mm vs. (60.12 ± 4.30) mm, t = 9.47, P < 0.001], and higher levels of left ventricular ejection fraction [(54.08 ± 3.06)% vs. (47.50 ± 3.22)%, t = 10.36, P < 0.001] compared with the control group. The levels of tumor necrosis factor-alpha, interleukin-6, and C-reactive protein in the observation group were significantly lower than those in the control group ( t = 5.53, 12.48, 13.45, all P < 0.001). Conclusion:Amlodipine/benazepril tablets are effective in facilitating the recovery of cardiac and vascular endothelial function in older adult patients with hypertension complicated by coronary heart disease. Furthermore, they improve blood pressure control and mitigate the inflammatory response in the body.

TNF-α promoter hypomethylation is frequent in oncopediatric patients who recovered from mucositis

Abstract The aim of this study was to investigate the DNA methylation profile in genes encoding catalase (CAT) and superoxide dismutase (SOD3) enzymes, which are involved in oxidative stress mechanisms, and in genes encoding pro-inflammatory cytokines interleukin-6 (IL6) and tumor necrosis factor-alpha (TNF-α) in the oral mucosa of oncopediatric patients treated with methotrexate (MTX®). This was a cross-sectional observational study and the population comprised healthy dental patients (n = 21) and those with hematological malignancies (n = 64) aged between 5 and 19 years. Oral conditions were evaluated using the Oral Assessment Guide and participants were divided into 4 groups: 1- healthy individuals; 2- oncopediatric patients without mucositis; 3- oncopediatric patients with mucositis; 4- oncopediatric patients who had recovered from mucositis. Methylation of DNA from oral mucosal cells was evaluated using the Methylation-Specific PCR technique (MSP). For CAT, the partially methylated profile was the most frequent and for SOD3 and IL6, the hypermethylated profile was the most frequent, with no differences between groups. For TNF-α, the hypomethylated profile was more frequent in the group of patients who had recovered from mucositis. It was concluded that the methylation profiles of CAT, SOD3, and IL6 are common profiles for oral cells of children and adolescents and have no association with oral mucositis or exposure to chemotherapy with MTX®. Hypomethylation of TNF-α is associated with oral mucosal recovery in oncopediatric patients who developed oral mucositis during chemotherapy.

Diagnostic accuracy of interleukin-6, interleukin-10 and tumor necrosis factor alpha cytokine levels in patients with mild cognitive impairment: systematic review and meta-analysis / Precisão diagnóstica dos níveis das citocinas IL-6, IL-10 e TNF-α em pacientes com comprometimento cognitivo leve: revisão sistemática e metanálise

ABSTRACT There is growing evidence suggesting an association between neurodegeneration and inflammation playing a role in the pathogenesis of age-associated diseases, including Alzheimer's disease (AD) and Mild Cognitive Impairment (MCI). Objective: A systematic review and meta-analysis were performed to verify evidence on the diagnostic accuracy parameters of the inflammatory cytokines interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor alpha (TNF-α). Methods: A search of Medical Literature Analysis and Retrieval System Online (Medline), Scientific Electronic Library Online (SciELO), Web of Science and Science Direct databases was performed and nine observational studies associated with peripheral inflammatory biomarkers in MCI were identified. Mean (±standard deviation — SD) concentrations of these biomarkers and values of true positives, true negatives, false positives and false negatives for MCI and healthy controls (HC) were extracted from these studies. Results: Significantly higher levels of IL-10 were observed in subjects in the MCI group and Mini-Mental State Examination (MMSE) scores were lower compared to HC. For the other investigations, no differences were found between the groups. Our meta-analysis for the TNF-α biomarker revealed high heterogeneity between studies in terms of sensitivity and specificity. Conclusion: These findings do not support the involvement of inflammatory biomarkers for detection of MCI, although significant heterogeneity was observed. More studies are needed to evaluate the role of these cytokines in MCI, as well as in other stages of cognitive decline and all-cause dementias.

RESUMO Há evidências crescentes que sugerem uma associação entre a neurodegeneração e a inflamação, desempenhando um papel na patogênese de doenças associadas à idade, incluindo a doença de Alzheimer (DA) e o comprometimento cognitivo leve (CCL). Objetivo: Uma revisão sistemática e metanálise foram realizadas para verificar evidências relativas aos parâmetros de acurácia diagnóstica das citocinas inflamatórias interleucina-6 (IL-6), interleucina-10 (IL-10) e fator de necrose tumoral (TNF-α). Métodos: Foi realizada uma busca nas bases de dados Medical Literature Analysis and Retrieval System Online (Medline), Scientific Electronic Library Online (SciELO), Web of Science e Science Direct, e foram identificados nove estudos observacionais associados a biomarcadores inflamatórios periféricos no CCL. As concentrações médias (desvio padrão — ±DP) desses biomarcadores e valores de verdadeiros positivos, verdadeiros negativos, falsos positivos e falsos negativos para CCL e controles saudáveis (CS) foram extraídos desses estudos. Resultados: Níveis significativamente mais elevados de IL-10 foram observados em indivíduos do grupo CCL e os escores do Miniexame do Estado Mental foram mais baixos em comparação com o CS. Para as demais investigações não foram encontradas diferenças entre os grupos. Nossa metanálise para o biomarcador TNF-α revelou alta heterogeneidade entre os estudos em termos de sensibilidade e especificidade. Conclusão: Esses achados não apoiam o envolvimento de biomarcadores inflamatórios na detecção do CCL, embora tenha sido observada heterogeneidade significativa. Mais estudos são necessários para avaliar o papel dessas citocinas no CCL, bem como em outros estágios de declínio cognitivo e demências de todas as causas.

Hepatitis B reactivation risk and physician awareness in rheumatological patients receiving anti-tumor necrosis factor-α treatment

SUMMARY OBJECTIVE: We aimed to evaluate the risk of hepatitis B virus reactivation in rheumatic patients using anti-tumor necrosis factor-alpha drugs and the awareness of physicians about hepatitis B virus reactivation. METHODS: Demographic characteristics, pre- and post-treatment hepatitis markers, and laboratory parameters of patients receiving anti-tumor necrosis factor-alpha therapy in our rheumatology clinic were retrospectively examined. RESULTS: A total of 448 patients, 240 (53.6%) female and 208 (46.4%) male, were evaluated. Their mean age was 48.02±14.64 years. While HBsAg was examined in 443 (98.9%) patients before treatment, 7 (1.6%) patients were found to be HBsAg positive. While anti-HBc IgG was examined in 405 (90.4%) patients, it was positive in 69 (17%) patients. HBs Ag (total 446-99.6%) test was performed in three patients who were not tested for HBsAg before the treatment, and anti-HBc total (431-96.2% total) test was performed in 26 patients who were not tested for anti-HBc total. All HBsAg positive patients and 17 (24.6%) of those with previous hepatitis B received antiviral treatment. While the median follow-up period of the patients was 24 (6-60) months, no patient developed hepatitis B virus reactivation. CONCLUSION: The screening rates and awareness of physicians providing anti-tumor necrosis factor-alpha therapy for hepatitis B virus infection were found to be higher compared to similar studies. Hepatitis B virus reactivation did not develop in any patient. Since the risk of hepatitis B virus reactivation is low, especially in patients with previous hepatitis B, it would be more appropriate to follow up the patients without giving antiviral prophylaxis.

New immunological aspects of peri-implantitis

ABSTRACT Objective: This study aimed to compare the levels of HIF1-α, VEGF, TNF-α, and IL-10 in the peri-implant crevicular fluid of patients with and without peri-implantitis. Methods: Forty patients, comprising 16 with and 24 without peri-implantitis were selected. Results: Patients with peri-implantitis exhibited significantly higher HIF-1α levels than those without peri-implantitis (p=0.0005). TNF-α revealed significant positive correlations with IL-10 (p=0.0008) and VEGF (p=0.0246), whereas HIF-1α and IL-10 levels (p=0.0041) demonstrated a negative and significative correlation in the peri-implantitis group. Conclusion: This study, for the first time demonstrates the balance of HIF-1α, TNFα, IL-10, and VEGF in peri-implantitis. It shows an elevated HIF-1α levels in patients with peri-implantitis, which could have stemmed from persistent inflammation- triggered hypoxia. Furthermore, the positive correlation between TNF-α and VEGF suggests intensified proinflammatory activity in peri-implantitis. Nevertheless, further studies are essential to understand these immune dynamics in peri-implantitis.

Hepatic and skeletal muscle tissue immunometabolism in trained rats / Inmunometabolismo del tejido hepático y muscular esquelético de ratas entrenadas / Imunometabolismo tecidual hepático e muscular esquelético de ratos treinados

ABSTRACT Introduction: Physical exercise can be an alternative for preventing and treating the harmful effects of obesity, mainly inflammatory effects on skeletal muscle and liver tissues. However, no consensus exists regarding this purpose's best physical training model. Objective: Evaluate morphological, metabolic, and inflammatory alterations in rats' skeletal and hepatic muscle tissues caused by aerobic and resistance training. Methods: 24 Wistar rats were divided into sedentary (S), aerobic (AE), and resistance training (R) groups. Blood glucose, total cholesterol, and serum triglycerides were measured periodically. After euthanasia, body mass was measured to calculate the total mass gain during the experiment. High-density lipoprotein (HDL) was measured. Adipose tissue was extracted to calculate its percentage relative to body mass and the liver, soleus, and gastrocnemius muscles for morphological analyses and concentrations of glycogen, lipids, and Tumor Necrosis Factor α (TNF-α). The Kruskall-Wallis test and Dunn's post-test were performed for statistical analysis, adopting p<0.05. Results: Both training models reduced the percentage of adipose tissue, body mass gain, and hepatic TNF-α concentration (p<0.05). AE increased serum HDL, gastrocnemius fiber diameter and reduced the fractal dimension in the soleus (p<0.05). R reduced blood glucose and serum and liver lipids, increased liver and soleus glycogen concentrations, increased gastrocnemius fiber diameter, and decreased TNF-α (p<0.05). Conclusion: Both training models reduced body mass, relative visceral adipose tissue, serum total cholesterol concentration, and liver inflammation. However, resistance training was more effective in promoting metabolic effects in the liver and skeletal muscle and reducing muscle inflammation in rats. Level of Evidence V; Expert Opinion.

RESUMEN Introducción: El ejercicio físico puede ser una alternativa para prevenir y tratar los efectos nocivos de la obesidad, principalmente los efectos inflamatorios sobre los tejidos del músculo esquelético y del hígado. Sin embargo, no existe consenso sobre cuál es el mejor modelo de entrenamiento físico para este fin. Objetivo: Evaluar las alteraciones morfológicas, metabólicas e inflamatorias del entrenamiento aeróbico y de resistencia en sobre los tejidos músculo esqueléticos y hepáticos de ratas. Métodos: 24 ratas Wistar se dividieron en grupos sedentarios (S), aeróbicos (AE) y de entrenamiento de resistencia (R). Se midieron periódicamente glucosa en sangre, colesterol total y triglicéridos. Después de la eutanasia, se midió la masa corporal para calcular la ganancia de masa total durante el experimento. Se midió la lipoproteína de alta densidad (HDL). Se extrajo tejido adiposo para calcular su porcentaje relativo a la masa corporal, así como hígado, músculos sóleo y gastrocnemio para análisis morfológicos y concentraciones de glucógeno, lípidos y Factor de Necrosis Tumoral α (TNF-α). Para el análisis estadístico fueron utilizados Kruskall-Wallis y el post-test de Dunn, adoptando p<0,05. Resultados: Ambos entrenamientos redujeron el porcentaje de tejido adiposo, masa corporal y la concentración de TNF-α hepático (p<0,05). AE aumentó el HDL sérico, el diámetro de la fibra del gastrocnemio y redujo la dimensión fractal en el sóleo (p<0,05). R redujo la glucosa en sangre y los lípidos séricos y hepáticos, aumentó las concentraciones de glucógeno hepático y sóleo, aumentó el diámetro de la fibra del gastrocnemio y disminuyó el TNF-α (p<0,05). Conclusión: Ambos modelos de entrenamiento redujeron la masa corporal, el tejido adiposo visceral relativo, la concentración sérica de colesterol total y la inflamación hepática. El entrenamiento de resistencia demostró ser más eficaz para promover los efectos metabólicos en el hígado y el músculo esquelético, además de reducir la inflamación muscular en ratas. Nivel de Evidencia V; Opinión del Especialista.

RESUMO Introdução: O exercício físico pode se apresentar como uma alternativa para prevenção e tratamento de efeitos deletérios da obesidade, principalmente efeitos inflamatórios sobre os tecidos muscular esquelético e hepático. No entanto, não há consenso quanto ao melhor modelo de treinamento físico para tal finalidade. Objetivos: Avaliar alterações morfológicas, metabólicas e inflamatórias dos treinamentos aeróbico e resistido sobre os tecidos muscular esquelético e hepático de ratos. Métodos: 24 ratos Wistar foram divididos nos grupos sedentário (S), treinamento aeróbico (AE) e resistido (R). Glicemia, colesterol total e triglicerídeos séricos foram mensurados periodicamente. Após a eutanásia, a massa corporal foi mensurada para calcular o ganho total de massa durante o experimento. A lipoproteína de alta densidade (HDL) foi dosada. O tecido adiposo foi extraído para cálculo de sua porcentagem relativa à massa corporal assim como o fígado e os músculos sóleo e gastrocnêmio para as análises morfológicas e das concentrações de glicogênio, lipídios e Fator de Necrose Tumoral α (TNF-α). Para análise estatística, foram utilizados o teste de Kruskall-Wallis e o pós-teste de Dunn, adotando-se p<0,05. Resultados: Ambos os modelos de treinamento reduziram o percentual de tecido adiposo, ganho de massa corporal e concentração hepática de TNF-α (p<0,05). AE aumentou o HDL sérico, o diâmetro das fibras do gastrocnêmio e reduziu a dimensão fractal no sóleo (p<0,05). R reduziu a glicemia e os lipídios séricos e hepáticos, aumentou a concentração de glicogênio hepático e sóleo, aumentou o diâmetro das fibras gastrocnêmicas e diminuiu o TNF-α (p<0,05). Conclusão: Ambos os modelos de treinamento reduziram a massa corporal, o tecido adiposo visceral relativo, a concentração sérica de colesterol total e a inflamação hepática. No entanto, o treinamento resistido mostrou-se mais eficaz em promover efeitos metabólicos no fígado e no músculo esquelético, além de reduzir a inflamação muscular em ratos. Nível de Evidência V; Opinião do Especialista.

Comparative Analysis of TNF-alpha, TNF-R1, and TNF-R2 in Patients with Low-impact Fractures Due to Osteoporosis / Análise comparativa de TNF-alfa, TNF-R1 e TNF-R2 em pacientes com fraturas de baixo impacto decorrentes de osteoporose

Abstract Objective To analyze the serum levels of TNF-alpha and its TNF-R1 and TNF-R2 receptors in the blood of patients with low-impact fractures due to osteoporosis, comparing between genders and with healthy patients. Methods The present study was conducted with a blood sample of 62 patients, divided into patients with osteoporosis and healthy patients. The results were obtained using the ELISA method. Cytokine concentrations were determined based on the absorbance values obtained. Results Serum TNF-alpha levels were undetectable in female patients, while in males they were found only in one patient, with no significant difference. Similar results were found in the analyses of TNF-R1 and TNF-R2 levels, a significant increase in levels of TNF-alpha receptors in the groups of patients with osteoporosis compared with the control groupinbothsexes.There wasnosignificant difference between the sexes in the dosage of both receptors within the group with osteoporosis. There was also a positive and significant correlation in the levels of TNF-R1 and TNF-R2 only in women. Conclusion The significant increase in TNF-R1 and TNF-R2 levels in women with osteoporosis suggest that the release and expression of these receptors may be contributing differently to the development of osteoporosis in men and women.

Resumo Objetivo Analisar os níveis séricos de TNF-alfa e de seus receptores TNF-R1 e TNF-R2 no sangue de pacientes com fraturas de baixo impacto, decorrentes de osteoporose, comparando entre os sexos e com pacientes saudáveis. Métodos Oestudofoi realizadocom amostradesanguede 62 pacientes,divididos em pacientes com osteoporose e pacientes saudáveis. Os resultados foram obtidos através do método de ELISA. As concentrações de citocinas foram determinadas com base nos valores de absorbância obtidos. Resultados Os níveis séricos de TNF-alfa foram indetectáveis nos pacientes do sexo feminino, enquanto no masculino encontrou-se somente em um paciente, não havendo diferença significativa. Encontrou-se resultados semelhantes nas análises dos níveis de TNF-R1 e TNF-R2, aumento significativo nos níveis dos receptores de TNF-alfa nos grupos de pacientes com osteoporose em comparação com o grupo controle, em ambos os sexos. Não houve diferença significativa entre os sexos na dosagem de ambos os receptores dentro do grupo com osteoporose. Houve ainda correlação positiva e significativa nos níveis de TNF-R1 e TNF-R2 apenas nas mulheres. Conclusão O aumento significativo nos níveis de TNF-R1 e TNF-R2 em mulheres com osteoporose sugerem que a liberação e expressão destes receptores pode estar contribuindo de maneira distinta no desenvolvimento da osteoporose em homens e mulheres.