Acute phase reactions in Daboia siamensis venom and fraction-induced acute kidney injury: the role of oxidative stress and inflammatory pathways in in vivo rabbit and ex vivo rabbit kidney models

Abstract Background: This study examines the direct nephrotoxic effects of Daboia siamensis venom (RVV) and venom fractions in in vivo and isolated perfused kidneys (IPK) to understand the role of inflammation pathways and susceptibility to oxidative stress in venom or fraction-induced acute renal failure. Methods: We administered RVV and its venom fractions (PLA2, MP, LAAO, and PDE) to rabbits in vivo and in the IPK model. We measured oxidative stress biomarkers (SOD, CAT, GSH, and MDA) in kidney tissue, as well as inflammatory cytokines (TNF-α, IL-1β, IFN-γ, IL-4, IL-5, and IL-10), MDA and GSH levels in plasma and urine. We also calculated fractional excretion (FE) for pro-/anti-inflammatory cytokines and oxidative stress biomarkers, including the ratios of pro-/anti-inflammatory cytokines in urine after envenomation. Results: In both kidney models, significant increases in MDA, SOD, CAT, and GSH levels were observed in kidney tissues, along with elevated concentrations of MDA and GSH in plasma and urine after injecting RVV and venom fractions. Moreover, RVV injections led to progressive increases in FEMDA and decreases in FEGSH. The concentrations of IL-4, IL-5, IL-10, IFN-γ, and TNF-α in plasma increased in vivo, as well as in the urine of the IPK model, but not for IL-1β in both plasma and urine after RVV administrations. Urinary fractional excretion of TNF-α, IL-1β, IFN-γ, IL-4, IL-5, and IL-10 tended to decrease in vivo but showed elevated levels in the IPK model. A single RVV injection in vivo disrupted the balance of urinary cytokines, significantly reducing either the TNF-α/IL-10 ratio or the IFN-γ/IL-10 ratio. Conclusion: RVV induces renal tubular toxicity by increasing oxidative stress production and elevating inflammatory cytokines in urine. During the acute phase of acute kidney injury, the balance of urine cytokines shifts toward anti-inflammatory dominance within the first two hours post-RVV and venom fractions.

Clinical Efficacy of Modified Banxia Xiexintang Combined with Vedolizumab in Treatment of Crohn's Disease with Syndrome of Cold and Heat in Complexity / 中国实验方剂学杂志

ObjectiveTo evaluate the clinical efficacy of modified Banxia Xiexintang combined with vedolizumab （VDZ） in the treatment of active moderate to severe Crohn's disease （CD） with the syndrome of cold and heat in complexity and the effect of the therapy on intestinal flora. MethodEighty patients were randomized based on the random number table method into a control group （40 cases） and an observation group （40 cases）. The control group was treated with VDZ， and the observation group was treated with modified Banxia Xiexintang （1 bag per day） combined with VDZ. The treatment in both groups lasted for 14 weeks and the follow-up lasted until the 52th weeks. The CD activity index （CDAI）， CD simplified endoscopic score （SES-CD）， inflammatory bowel disease questionnaire （IBDQ） score， and syndrome score of cold and heat in complexity were assessed before treatment， after treatment， and at the end of follow-up. The levels of hemoglobin （HGB）， hematocrit （HCT）， albumin （ALB）， C-reactive protein （CRP）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-17 （IL-17）， and fecal calprotectin （FC） were measured before and after treatment. Intestinal flora was examined before and after treatment. The safety of the therapy was evaluated. ResultCompared with those before treatment， the scores of CDAI， SES-CD， and the syndrome of cold and heat in complexity decreased （P<0.05） and the IBDQ score increased after treatment （P<0.05）. Compared with those before treatment， the scores of CDAI， SES-CD， and the syndrome of cold and heat in complexity increased （P<0.05） and the IBDQ score decreased （P<0.05） at the end of follow-up. After treatment and at the end of follow-up， the observation group had lower scores of CDAI， SES-CD， and syndrome of cold and heat （P<0.05） and higher IBDQ score （P<0.05） than the control group. Moreover， the observation group had higher clinical remission rate（χ2=4.381，3.962） and response rate（χ2=5.541，4.306） and lower non-response rate（χ2=6.646，4.306） than the control group at the two time points （P<0.05）. The endoscopic remission rate（χ2=4.072，3.985） and response rate（χ2=4.528，5.161） in the observation group were higher than those in the control group （P<0.05）. After treatment， the HGB， HCT， and ALB levels in both groups elevated， and the observation group had higher levels than the control group （P<0.05）. The treatment in both groups lowered the levels of CRP， IL-6， TNF-α， IL-17， and FC （P<0.05）， and the observation group had lower levels of CRP， IL-6， TNF-α， IL-17， and FC than the control group after treatment （P<0.05）. The relative abundance of Bifidobacterium， Lactobacillus， and Prevotella increased （P<0.05）， while that of Proteus， Klebsiella， and Enterococcus decreased （P<0.05） in the two groups after treatment. Moreover， the changes in the relative abundance of these bacteria in the observation group were more obvious than those in the control group （P<0.05）. No adverse reactions related to the modified Modified Banxia Xiexintang were observed during the study period. ConclusionModified Banxia Xiexintang combined with VDZ can play a synergistic role and has good short-term and long-term efficacy. This therapy can improve the nutritional status， regulate intestinal flora， and reduce inflammatory injury in the treatment of moderate to severe active CD patients with the syndrome of cold and heat in complexity without causing severe adverse reactions.

Preparation of tripterygium glycoside nanoparticles and therapeutic effect on arthritis rats / 中国药理学通报

Aim To prepare tripterygium glycoside nanoparticles and probe into their therapeutic effect on collagen-induced arthritis ( CIA) rats. Methods Tripterygium glycosides polyglycoside nanoparticles were prepared by thin film dispersion method and their quality was assessed. The CIA model was established and drug intervention performed. The body weight, toe swelling degree and arthritis index were measured. The pathological changes of the organs, knee and ankle synovium were observed. The serum levels of kidney function and inflammatory cytokine expression were detected in rats. Results The prepared tripterygium wil-fordii polyglycoside nanoparticles were round particles with uniform distribution and stable properties under electron microscope. Compared with the model group, the swelling of the left and right toes of medication group significantly decreased (P < 0. 01), and the ar-thritis index markedly decreased ( P < 0. 01). Among them, the efficacy of the TG-NPs group was better than that of the TG group. Compared with the normal group, the indexes of heart, spleen, kidney and testis all significantly decreased (P <0. 05, P<0.01). TG-NPs group had a significantly reduced pathological ankle-joint injury in knee cartilage and increased apoptotic synovial cells. Compared with the model group, the serum levels of ALT and BUN and CRE in TG-NPs group were significantly lower (P < 0. 05 ), and IL-1β, TNF-α and IL-6 levels decreased significantly (P <0. 05). Conclusions TG-NPs have good therapeutic effect on CIA through induction of synovial cell apoptosis and decrease of the expression of inflammatory cytokines. By intravenous injection of blood circula-tion, slow and controlled release of drugs can be achieved, the first pass effect caused by oral drug can be avoided, the viscera toxicity can be reduced, which provides an experimental basis for the development of new nanoagents for the treatment of rheumatoid arthritis.

Antitumor and immunomodulatory activity of fucoidan from the brown alga Lessonia trabeculata (Lessoniaceae) on breast cancer spheroids / Actividad antitumoral e inmunomoduladora de fucoidan del alga parda Lessonia trabeculata (Lessoniaceae) en esferoides de cáncer de mama

Abstract Introduction: The therapeutic benefits of the brown algae fucoidan in the treatment of breast cancer have attracted considerable interest in recent years. However, research using spheroids which provide relevant results in trials for antitumor and immunomodulatory products because they adequately simulate the tumor microenvironment, is limited. Objective: To evaluate the antitumor and immunomodulatory activity of Lessonia trabeculata fucoidan (LtF), native to the Peruvian Sea, on two types of multicellular tumor spheroids. Methods: The study was conducted from January to December 2021. Two types of spheroides were elaborated: from 4T1 tumor cells (MTS), and from 4T1 tumor cells+mouse splenocytes (MTSs). The antitumor activity of LtF was evaluated in MTS by quantifying cell viability with MTT. Immunomodulatory activity was determined in MTSs using the IC50 for two types of treatment: simple, fucoidan alone (LtF) and combined, fucoidan+doxorubicin (LtF+Dox). Pro-inflammatory (TNF-α, IL-6) and anti-inflammatory (IL-10, TGF-β) cytokine production was quantified by sandwich ELISA 72 h after treatment. Dox was used as positive control in all assays. Results: LtF exerted antitumor activity as evidenced by increased necrotic zone and cell debris formation compared to the untreated control. Antitumor activity was concentration dependent between 100 and 6 000 μg/ml. In MTSs, simple treatment increased IL-6 and decreased IL-10 and TGF-β production. The combined treatment significantly reduced TGF-β production. In both treatments and Dox, there was an increase in IL-6 compared to the untreated control. The highest production of IL-10 and TGF-β was observed in the untreated control, compatible with a highly immunosuppressive tumor microenvironment. Conclusions: LtF is a good candidate for the treatment of breast cancer and can immunomodulate the tumor microenvironment alone or in combination with Dox.

Resumen Introduccción: Los beneficios terapéuticos del fucoidan de algas pardas en el tratamiento del cáncer de mama han despertado gran interés en los últimos años. Sin embargo, las investigaciones con esferoides son limitadas, éstos proporcionan resultados relevantes en ensayos de productos antitumorales e inmunomoduladores porque simulan adecuadamente el microambiente tumoral. Objetivo: Evaluar la actividad antitumoral e inmunomoduladora del fucoidan de Lessonia trabeculata (LtF), nativa del Mar Peruano, en dos tipos de esferoides tumorales multicelulares. Métodos: El estudio se realizó de enero a diciembre de 2021. Se elaboraron dos tipos de esferoides: con células tumorales 4T1 (MTS) y con células tumorales 4T1+esplenocitos de ratón (MTSs). La actividad antitumoral de LtF se evaluó en MTS cuantificando la viabilidad celular con MTT. La inmunomodulación se determinó en MTSs utilizando la IC50 para dos tipos de tratamiento: simple, fucoidan solo (LtF) y combinado, fucoidan+doxorubicina (LtF+Dox). La producción de citoquinas proinflamatorias (TNF-α, IL-6) y antiinflamatorias (IL-10, TGF-β) se cuantificó mediante ELISA sándwich 72 h post-tratamiento. En todos los ensayos se utilizó Dox como control positivo. Resultados: En los MTS, el LtF ejerció actividad antitumoral evidenciada por aumento de la zona necrótica y formación de restos celulares respecto al control no tratado. La actividad antitumoral fue concentración-dependiente entre 100 y 6 000 μg/ml. En los MTSs, con el tratamiento simple se incrementó IL-6 y disminuyeron IL-10 y TGF-β. El tratamiento combinado redujo significativamente la producción de TGF-β. Los dos tratamientos y Dox incrementaron IL-6 respecto al control no tratado. La mayor producción de IL-10 y TGF-β se observó en los no tratados, compatible con un microambiente tumoral altamente inmunosupresor. Conclusiones: El LtF es un buen candidato para tratar el cáncer de mama y puede inmunomodular el microambiente tumoral solo o en combinación con Dox.

Mecanismos y dianas terapéuticas del inflamasoma NLRP3 en la miocardiopatía diabética / NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy

Resumen La cardiomiopatía diabética es una complicación grave de la diabetes causada por estrés oxidativo, inflamación, resistencia a la insulina, fibrosis miocárdica y lipotoxicidad. Se trata de un padecimiento insidioso, complejo y difícil de tratar. El inflamasoma NLRP3 desencadena la maduración y liberación de citoquinas proinflamatorias, participa en procesos fisiopatológicos como la resistencia a la insulina y la fibrosis miocárdica, además de estar estrechamente relacionado con la aparición y progresión de la cardiomiopatía diabética. El desarrollo de inhibidores dirigidos a aspectos específicos de la inflamación sugiere que el inflamasoma NLRP3 puede utilizarse para tratar la cardiomiopatía diabética. Este artículo pretende resumir el mecanismo y las dianas terapéuticas del inflamasoma NLRP3 en la cardiomiopatía diabética, así como aportar nuevas sugerencias para el tratamiento de esta enfermedad.

Abstract Diabetic cardiomyopathy (DCM) is a serious complication of diabetes caused by oxidative stress, inflammation, insulin resistance, myocardial fibrosis, and lipotoxicity; its nature is insidious, complex and difficult to treat. NLRP3 inflammasome triggers the maturation and release of pro-inflammatory cytokines, participates in pathophysiological processes such as insulin resistance and myocardial fibrosis, in addition to being closely related to the development and progression of diabetic cardiomyopathy. The development of inhibitors targeting specific aspects of inflammation suggests that NLRP3 inflammasome can be used to treat diabetic cardiomyopathy. This paper aims to summarize NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy, and to provide new suggestions for the treatment of this disease.

The role of pro-inflammatory cytokines – tumor necrosis factor-alpha and interleukin 6 – in the evolution of diabetic nephropathy: A comparative study

Background: Diabetes is highly prevalent and it is responsible for the increased financial burden on healthcare. Type II diabetes is a more prevalent form of diabetes. Uncontrolled and unsupervised type II diabetes may lead to various microvascular and macrovascular complications which are responsible for high morbidity and mortality. Diabetic nephropathy (DN) is a common complication characterized by the expansion of mesangial cells with thickening of the basement and nodular glomerulosis. TNF-alpha and IL-6 play an important role in causing detrimental changes leading to nephropathy. The study of the role of these inflammatory cytokines in patients with DN may help in the early diagnosis and management. Aims and Objectives: The objectives of this study were to compare the levels of proinflammatory cytokines, TNF-?, and IL-6 in the evolution of DN patients. Materials and Methods: The present study was conducted in the Department of Biochemistry, in collaboration with the Department of Medicine (Nephrology unit); Pt. B.D. Sharma, Post Graduate Institute of Medical Sciences, Rohtak after ethical clearance. Forty patients with DN (Stages 3, 4, and 5) and forty patients with diabetes mellitus without nephropathy were taken up for study after taking informed consent. Results: The mean serum TNF-? levels in cases was 33.05 ± 29.22 pg/mL and in controls was 17.67 ± 12.33 pg/mL. On the basis of unpaired t-test, the difference between the groups was statistically highly significant (P < 0.05). The mean serum interleukin-6 levels in cases was 24.92 ± 30.16 pg/mL (2.95–155.55 pg/mL) and in controls was 6.76 ± 5.82 pg/mL (2.22–35.42 pg/mL). On the basis of the t-test, the difference between the groups was statistically highly significant (P < 0.05). Conclusion: TNF-? and IL-6 may serve as potential biomarkers for patients with DN and also in the development of newer therapeutic modalities for the prevention and treatment of DN.

Anti-inflammatory and antioxidant potential of Artemisia dracunculus L. aqueousextract against acetic acid induced ulcerative colitis in male Wistar rats

Ulcerative colitis (UC) is one of the two types of inflammatory bowel disease (IBD) which is increasing worldover due to modern life style. Patients with UC are prone to develop colorectal cancer. While the disease severity decides the treatment option, researchers look towards herbal medicines with anti-inflammatory properties for minimal or nil side effects. Artemisia dracunculus L., commonly called Tarragon, is a medicinal herb used in traditional Asian medicine mainly in Iran, India, Pakistan and Azerbaijan due to its special compounds. In this study, we tried to elucidate the effects of aqueous extract of tarragon on acetic acid induced ulcerative colitis (UC) in rats. Male Wistar rats were grouped into four groups of eight each viz., control; experimental control (UC was induced via luminal instillation of 4% acetic acid); and UC induced + aqueous tarragon extract (100 mg/kg) or prednisolone (2 mg/kg) orally for ten consecutive days. Tissue specimens were collected after the experimental period for evaluation of caspase-3 and cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Real-time PCR was used to monitor the levels of IL-1, IL-6 and TNF-? in colonic homogenates. Moreover, the levels of myeloperoxidase, nitric oxide and total antioxidant capacity were measured in colonic homogenates. The results showed that both treatment regimens could similarly reduce the severity of disease symptoms. Treatment with aqueous extract of tarragon caused a better improvement (P <0.05) in the levels of myeloperoxidase enzyme, and total antioxidant capacity of colonic homogenates compared to prednisolone. Nevertheless, the levels of the expression of caspase-3, and COX-2 and TNF-? were reduced in UC rats received prednisolone more than UC rats received aqueous extract of tarragon. The was no statistical difference in the levels of nitric oxide, IL-1 and IL-6 between UC rats received tarragon extract or prednisolone. Overall, these findings suggest that the aqueous extract of tarragon is a promising strategy to control ulcerative colitis. Aqueous extract can also be used as an anti-inflammatory and immune system stimulant in conditions where the immune system is damaged.

Chinese Medicine Regulates Knee Osteoarthritis-related Signaling Pathways： A Review / 中国实验方剂学杂志

As the research of traditional Chinese medicine （TCM） on knee osteoarthritis （KOA） is progressing， researchers have discovered that a variety of Chinese medicines can delay the progress of KOA by regulating signaling pathways at the molecular level. The Chinese medicines and their active ingredients mentioned in this article are associated with the signaling pathways in KOA. They can regulate the levels of targeted molecules via different signaling pathways to inhibit cartilage inflammatory cytokine， apoptosis， and cartilage matrix degradation and promote chondrocyte autophagy， so as to reduce the synovial inflammatory edema and delay cartilage degeneration. This paper systematically reviews the studies about the TCM intervention of KOA. Baicalein can reduce the inflammatory cytokines and apoptosis and promote the autophagy of chondrocytes by blocking the phosphatidylinositol-3 kinase/protein kinase （PI3K/Akt） signaling pathway. Cornuside I can decrease the phosphorylation activity of Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） pathway to reduce synovial inflammation and delay cartilage matrix degeneration. Salvianolic acid A can reduce inflammation and cartilage matrix degradation by inhibiting the phosphorylation of the nuclear factor-κB （NF-κB） pathway. Emodin can reduce the activity of Wnt/β-catenin pathway to inhibit the decomposition of collagen and proteoglycan. Myristicoside can inhibit apoptosis by blocking the p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway. Akebia saponin D can enhance the activity of nuclear factor E2-related factor 2/heme oxygenase 1（Nrf2/HO-1） pathway to inhibit oxidative stress in chondrocytes. The saponins in Achyranthis Bidentatae Radix reduce cartilage matrix degradation by enhancing the transforming growth factor-β （TGF-β）/Smad signaling pathway. Crocin inhibits the cartilage inflammation and apoptosis factor increase by stimulating the activity of hippo-Yes-associated protein （Hippo-YAP）. Ligustrazine blocks the Notch pathway to improve the morphology and abnormality of chondrocytes. Oleanolic acid reduces the destruction and degeneration of cartilage matrix via the estrogen signaling pathway. The above summary aims to provide references for future clinical and experimental research on KOA.

Effect of Modified Gualou Zhishitang Combined with Piperacillin Sodium and Tazobactam Sodium on Immune Function and Serum Levels of Inflammatory Cytokines in Patients with Stroke-associated Pneumonia with Syndrome of Phlegm-heat Accumulation in Lung / 中国实验方剂学杂志

ObjectiveTo explore the effects of modified Gualou Zhishitang combined with piperacillin sodium and tazobactam sodium on the immune function and serum levels of inflammatory cytokines in the patients with stroke-associated pneumonia （SAP， syndrome of phlegm-heat accumulation in lung）. MethodEighty SAP patients with the syndrome of phlegm-heat accumulation in lung were randomized into a control group （40 cases） and a study group （40 cases）. The SAP patients in the control group were treated with piperacillin sodium and tazobtam sodium， while those in the study group were treated with modified Gualou Zhishitang on the basis of the treatment in the control group for 2 consecutive weeks. The clinical therapeutic effects， immune function indexes， inflammation indexes， and lung function of SAP patients in the two groups before and after treatment were determined and compared. ResultAfter treatment， the scores of lesion， pulmonary rales， cough， fever， phlegm color， and constipation in both groups decreased （P<0.05）. After treatment， the ratio of forced expiratory volume in the first second to forced expiratory volume （FEV1/FVC） and forced expiratory volume in the first second as percentage of predicted value（FEV1%） in both groups improved （P<0.05）， and the study group outperformed the control group （P<0.05）. The treatment decreased the neutrophil to lymphocyte ratio （NLR） in the two groups （P<0.05）， and the study group had lower NLR than the control group after treatment （P<0.05）. The serum levels of procalcitonin （PCT） and hypersensitive C-reactive protein （hs-CRP） in both groups declined after treatment （P<0.05）， and the declines were more significant in the study group than in the control group （P<0.05）. After treatment， the study group was better than the control group （P<0.05）. The treatment in both groups elevated the levels of CD3+， CD4+， and CD4+/CD8+ in the peripheral blood and lowered the level of CD8+ （P<0.05）， and the changes were more significant in the study group than in the control group （P<0.05）. The total response rate of the study group was 95.00% （38/40）， which was higher than that （80.00%， 32/40） of the control group （χ2=4.114，P<0.05）. There was no significant difference in the incidence of adverse reactions between the two groups. ConclusionModified Gualhou Zhishitang combined with piperacillin sodium and tazobactam sodium demonstrates a significant therapeutic effect on the SAP patients with the syndrome of phlegm-heat accumulation in lung. This therapy can mitigate the clinical symptoms， improve the lung function， lower the serum levels of inflammatory cytokines， and improve the immune capacity， with high safety.

The correlation between brain functional network connectivity and inflammatory cytokines in patients with bipolar disorder II depressive episodes / 中华行为医学与脑科学杂志

Objective:To investigate the functional connectivity of default mode network (DMN) and limbic system, the expression level of inflammatory cytokine and their correlation in bipolar disorder type Ⅱ(BDⅡ) patients with depressive episodes.Methods:Thirty-three BD Ⅱ patients with depressive episodes and forty-six healthy controls were recruited to complete the resting-state functional magnetic resonance imaging (rs-fMRI). After image preprocessing, the DMN and limbic system were extracted from the image data by independent component analysis (ICA), so as to compare the differences of functional connectivity of resting brain network between the patients and the controls.Serum levels of inflammatory cytokines interleukin-6 (IL-6), interleukin-8(IL-8), interleukin-10(IL-10), tumor necrosis factor-α (TNF-α), and C-C motif chemokine ligand 4 (CCL4) in patients and healthy controls were detected.The correlation between functional connectivity of different brain regions and inflammatory cytokines was analyzed.SPSS 17.0 software was used for data statistical analysis.The two samples were compared using t-test or Mann-Whitney U-test, and Spearman was used for correlation testing. Results:In BDⅡ patients, the functional connectivity of the right medial prefrontal cortex(cluster-size=7 voxel, cluster-level PGRF<0.05, MNI: x=6, y=54, z=9, t=-3.765) and the left superior frontal gyrus(cluster-size=10 voxel, cluster-level PGRF<0.05, MNI: x=-21, y=54, z=15, t=-4.139) in DMN decreased, while the left cerebellum Ⅳ and Ⅴ lobules of limbic system (cluster-size=21 voxel, cluster-level PGRF<0.05, MNI: x=-15, y=-24, z=-30, t=4.468) and cerebellar tonsil of left cerebellum posterior lobe(cluster-size=8 voxel, cluster-level PGRF<0.05, MNI: x=-15, y=-51, z=-45, t=4.138) in the limbic system increased.Compared with the healthy controls, the serum levels of IL-10(7.39 (6.33, 9.32) pg/mL vs 6.54 (5.84, 7.39) pg/mL, Z=-2.937, P=0.003)and CCL4 (39.31 (25.77, 68.70) pg/mL vs 31.30 (20.32, 40.89) pg/mL, Z=-2.209, P=0.027) were higher in BDⅡ patients.The functional connectivity of the left cerebellum Ⅳ and Ⅴ lobules was positively correlated with the serum levels of IL-10 ( r=0.432, P=0.031) and that of the cerebellar tonsil of left cerebellum posterior lobe was positively correlated with the serum levels of IL-10 ( r=0.429, P=0.032) and CCL4 ( r=0.402, P=0.046). Conclusion:The functional connectivity of DMN and limbic system in BDⅡ patients with depressive episode is abnormal in resting-state fMRI.The expression level of inflammatory cytokines in patients' serum increases, and has correlation with the functional connection of limbic system.

The expression of miR-122 in the liver of rats infected with Clonorchis sinensis and its correlation with inflammatory cytokines / 中华地方病学杂志

Objective:To observe the expression changes of microRNA(miR)-122 in liver tissue of rats infected with Clonorchis sinensis and its correlation with expression level of inflammatory cytokines. Methods:Totally 24 SPF grade Wistar male rats were selected and randomly divided into a control group (100 μl physiological saline gavage), a 4-week infection group (100 Clonorchis sinensis metacercariae gavage), and an 8-week infection group (100 Clonorchis sinensis metacercariae gavage) based on body weight (100-120 g) using a random number table method, with 8 rats in each group. Starting from the third week of infection, rat feces were collected and directly smeared with physiological saline for identification of Wistar rat animal models infected with Clonorchis sinensis. After 4 and 8 weeks of infection, the rats in the 4- and 8-week infection groups were euthanized, while 4 rats in the control group were euthanized, respectively. The heart blood and left lobe liver tissue and serum samples were collected from each group of rats. Using hematoxylin-eosin (HE) staining to observe liver pathological damage under the light microscope, real-time fluorescence quantitative PCR to detect the expression level of miR-122 in liver tissue, and Luminex 200 liquid suspension chip to detect the expression levels of serum inflammatory cytokines [tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6 (IL-6)]. The correlation between miR-122 and inflammatory cytokines was analyzed using Pearson correlation. Results:Under the light microscope, the morphology of hepatocytes in control group was normal, and no inflammatory cell infiltration was observed. There was inflammatory cells such as lymphocyte, eosinophil and other inflammatory cell infiltration around the portal area in the 4-week infection group. The hepatocytes of the 8-week infected rats were arranged in a disordered manner, with varying degrees of swelling, loose and lightly stained cytoplasm, and some hepatocytes showed watery degeneration; additionally, bile duct dilation and thickening of the bile duct wall were observed in the liver tissue. There were statistically significant differences of liver miR-122 (1.00 ± 0.32, 2.57 ± 0.60, 3.63 ± 1.63), serum TNF-α [(0.14 ± 0.06), (0.43 ± 0.09), (0.61 ± 0.10) ng/ml], and IL-6 expression levels [(0.03 ± 0.01), (1.06 ± 0.24), (1.48 ± 0.33) ng/ml] in control group, 4- and 8-week infection groups ( F = 13.36, 69.99, 82.23, P < 0.001). There was no statistically significant difference in expression level of IL-1β between different groups ( F = 2.15, P = 0.141). The Pearson correlation analysis showed that the expression level of miR-122 was positively correlated with the expression levels of inflammatory cytokines TNF-α and IL-6 ( r = 0.67, 0.80, P < 0.001). Conclusion:Clonorchis sinensis infection can increase the expression of miR-122 in the host liver tissue, and the miR-122 is closely related to the expression levels of inflammatory cytokines TNF-α and IL-6.

Inhibitory effect of miR-497 on corneal neovascularization and its regulation mechanism targeting STAT3 / 中华实验眼科杂志

Objective:To investigate the role of microRNA (miR)-497 in the formation of corneal neovascularization (CNV) induced by alkali burn and its mechanism.Methods:Forty-two wild type (WT) C57BL/6 mice aged 6 to 8 weeks, 42 CRISPR/Cas9 mediated miR-497 knockout (KO) and 42 CRISPR/Cas9 mediated overexpression transgenic (TG) C57BL/6 mice were selected and assigned as WT group, KO group and TG group, respectively.The corneal alkali burn model was established.At 3, 7, 14 and 21 days after modeling, corneal epithelium damage and stromal turbidity were scored according to slit lamp microscopy.The area of neovascularization was measured.Corneal structural changes and expression of inflammatory cells were observed by histopathological staining.The expression of CD31 in corneal tissues was detected by immunohistochemistry staining.The targeted binding relationship between miR-497 and signal transducer and activator of transcription 3 (STAT3) was detected by luciferase reporter assay.The relative expressions of miR-497, vascular endothelial growth factor A (VEGFA), tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β and macrophage inflammatory protein (MCP)-1 mRNA were detected by real-time quantitative PCR.At 14 days following modeling, the expression of STAT3 and p-STAT3 proteins in mice corneal tissues was detected by Western blot.The use and care of animals complied with the ARVO statement.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.2019K-K010).Results:Corneal injury, inflammatory cell infiltration and CNV occurred in mice cornea after alkali burn.Corneal epithelial injury score, corneal stromal turbidity score and CNV area increased first and reached the peak on the 14th day after modeling, and then decreased.There were significant differences in corneal epithelial injury score, corneal stromal turbidity score, CNV area and number of CD31-positive cells among various time points after alkali burn ( Fgroup=49.19, 34.56, 44.56, 77.56; all at P<0.01; Ftime=51.62, 65.62, 71.32, 46.12; all at P<0.01). Corneal epithelial injury score, corneal stromal turbidity score, CNV area and the number of CD31-positive cells were greater in KO group at various time points than in WT and TG groups, and those in WT group were greater than in TG group (all at P<0.05). In WT STAT3 co-transfected cells, the luciferase activity of the miR-497 group was significantly lower than that of the miR-negative control group and normal control group (both at P<0.05). In mutant STAT3-transfected cells, there was no significant difference in luciferase activity among all groups ( F=0.69, P=0.56). On the 14th day after modeling, the relative expression levels of miR-497 in corneal tissue of WT, KO and TG groups were 0.68±0.11, 0.41±0.06 and 1.05±0.14, respectively, which were significantly lower than 1.00±0.04, 0.56±0.07 and 1.34±0.11 before modeling (all at P<0.01). The relative expressions of STAT3 and p-STAT3 were higher in KO group than in WT and TG groups, and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.05). The expressions of VEGFA, TNF-α, IL-6, IL-1β and MCP-1 mRNA at various time points after modeling in various groups were significantly higher than before modeling, which were higher in KO group than in WT and TG groups and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.01). Conclusions:MiR-497 inhibits corneal inflammation and CNV formation induced by alkali burn.It might inhibit the activation of the inflammation signal pathway via targeting STAT3.

Chlorogenic acid derived from Moringa oleifera leaf as a potential antiinflammatory agent against cryptosporidiosis in mice

@#Cryptosporidiosis is a serious illness in immunodeficient patients, and there is still no drug that can completely remove the parasite from the host. The present study represents the first report investigating the impact of the active molecule chlorogenic acid (CGA), naturally isolated from Moringa oleifera leaf extract (EMOLE), on immunosuppressed, Cryptosporidium parvum-infected BALB/c mice. Mice were divided into five groups: normal mice, infected immunosuppressed mice, and infected immunosuppressed mice treated with EMOLE, CGA, and nitazoxanide (NTZ) drugs. Parasitological, immunological, and histopathological investigations were recorded besides differences in the mice’ body weight. Infected control mice showed elevated levels of oocyst shedding throughout the study. The EMOLE- and CGA-treated groups showed 84.2% and 91.0% reductions in oocyst shedding, respectively, with no significant difference compared to the drug control. The inflammatory markers IFN-γ, IL-6, IL-1β, and TNF-α were significantly higher in the infected control group. Treatment with 300 mg/kg/day of EMOLE or 30 mg/kg/day of CGA significantly downregulated pro-inflammatory cytokine levels compared to the infected group, although they did not change significantly compared to the NTZ-treated group. Histopathology of intestinal sections showed inflammatory and pathological changes in the infected control group. Low-grade tissue changes and an obvious improvement in villi structure were seen in mice treated with CGA. This study highlighted the role of CGA, isolated and purified from EMOLE, as an effective anti-inflammatory agent in eradicating C. parvum infection.

The protective effect of cuminaldehyde on gastric mucosa in Rattus norregicus of experimental gastric ulcer / 中国药理学通报

Aim To explore the effect of cuminaldehyde in cumin fruit on gastric ulcer and the protective mechanism via establishing the gastric ulcer model of rats was by ethanol injury. Methods Thirty-six male R. norregicus were divided into six groups: control group, model group, omeprazole positive control group and cuminaldehyde low, medium and high dosage groups. After seven days of continuous intragastric administration, the acute gastric ulcer of R. norregicus was tested by absolute alcohol. Gastric ulcer area, inhibition rate, gastric tissue antioxidant activity, serum inflammatory factors and gastric mucosal protective factors were detected in different groups. Results The results showed that cuminaldehyde significantly reduced the area of gastric ulcer and increased the inhibition rate of gastric ulcer. The inhibition rate of cuminaldehyde at high dose group was up to 74.65%, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH) in gastric tissue significantly increased, and the contents of serum prolandin E

Mechanism of regulating immune dysfunction of sepsis based on network pharmacology and experimental verification / 中国药理学通报

Aim To verify the role of Shengjiang Powder in sepsis and explore its molecular mechanism. Methods The targets of drug active ingredients and disease-related targets were searched by TCMSP, Disgenet and other databases, and the intersection of the two was selected. DAVID database was used to carry out enrichment analysis of GO and KEGG pathways for intersection targets, and molecular docking was performed between drug active ingredients and core genes of key pathways. Mouse model of sepsis was constructed by cecal ligation puncture (CLP). Spleen tissue and serum of mice were collected. The percentage of T cell subsets in spleen was detected by flow cytometry, and IL-6 and IL-10 levels in serum were detected by ELISA. Results A total of 25 active ingredients, 238 targets of active ingredients, 2797 disease-related targets, 90 genes of intersection between active ingredients and disease-related targets, potential targets were AKT, JUN, EGFR, MMP9, etc. GO enrichment analysis showed 1021 items, including 942 biological processes, 23 cell compositions and 55 molecular functions. KEGG pathway analysis found that the intersection genes were mainly enriched in THE PD-1/PD-L1 signaling pathway, HIF-1 signaling pathway, TNF signaling pathway and inflammatory mediators signaling pathway, indicating that the therapeutic effect may be related to these pathways. The molecular docking results showed that quercetin, kamanol, emodin and other core compounds could be well combined with key genes. Flow cytometry results showed that after seven days of CLP, the proportion of CD4 T cells in spleen decreased, the proportion of CD4 PD-1 T cells increased, the release of IL-6 decreased, the content of IL-10 increased, and the mice were immunosuppressed. The percentage of CD4 T cells in spleen increased, the number of CD4 PD-1 T cells decreased, the release of IL-6 was enhanced, the content of IL-10 decreased, and the immunosuppression was improved. Conclusions It is proved that Shengjiang Powder can increase the release of pro-inflammatory cytokine IL-6, increase the ratio of CD4 T/CD8 T cells, and decrease the expression of anti-inflammatory cytokine IL-10 in the late stage of sepsis, so as to improve immune suppression in the late stage of sepsis and improve the survival rate of mice.

Short-Term Efficacy and Postoperative Inflammatory Cytokine Changes in Lung Cancer Treated with Da Vinci Robot-assisted Versus Video-assisted Thoracoscopic Surgery / 肿瘤防治研究

Objective To evaluate the short-term outcomes and postoperative inflammatory cytokine changes in patients with lung cancer treated with robot-assisted thoracoscopic surgery (RATS) versus video-assisted thoracoscopic surgery (VATS). Methods A total of 270 patients with lung cancer treated by minimally invasive surgery were selected for the study, and the surgical procedures were selected according to the patients' economic conditions and preferences. Among them, 132 patients completed the operation through RATS, and 138 patients completed the operation through VATS. The clinical data of the two groups were compared. Results All patients successfully completed radical lung cancer surgery, and no perioperative deaths were reported. Intraoperative bleeding, postoperative drainage time, postoperative hospital stay, number of lymph nodes dissected, and number of lymph nodes dissected groups were more advantageous in the RATS group compared with the VATS group (P < 0.05). In terms of operative time, total postoperative chest drainage, and hospitalization cost, the VATS group had an advantage (P < 0.05). The postoperative levels of CRP, PCT, IL-6, IL-8, IL-10, and TNF-α increased in both groups, compared with preoperative levels, and the increases in the RATS group were lower than those in the VATS group. Conclusion RATS offers technical and short-term efficacy advantages for the treatment of lung cancer but comes with the disadvantage of high cost. Post-operative inflammatory cytokine elevation is lower in the RATS group, and inflammatory response to the organism is less severe.

Clinical study of 19 cases of steroid-refractory gastrointestinal acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation with fecal microbiota transplantation / 中华血液学杂志

Objective: To investigate the clinical efficacy of fecal microbiota transplantation (FMT) for treating steroid-refractory gastrointestinal acute graft-versus-host disease (GI-aGVHD) . Methods: This analysis included 29 patients with hematology who developed steroid-refractory GI-aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Huaian Hospital Affiliated to Xuzhou Medical University from March 2017 to March 2022. Among them, 19 patients underwent FMT treatment (the FMT group) and 10 patients did not (the control group). The efficacy and safety of FMT were assessed, as well as the changes in intestinal microbiota abundance, lymphocyte subpopulation ratio, peripheral blood inflammatory cytokines, and GVHD biomarkers before and after FMT treatment. Results: ① Complete remission of clinical symptoms after FMT was achieved by 13 (68.4%) patients and 2 (20.0%) controls, with a statistically significant difference (P<0.05). Intestinal microbiota diversity increased and gradually recovered to normal levels after FMT and FMT-related infections did not occur. ②The proportion of CD3(+) and CD8(+) cells in the FMT group after treatment decreased compared with the control group, and the ratio of CD4(+), regulatory T cells (Treg), and CD4(+)/CD8(+) cells increased (all P< 0.05). The interleukin (IL) -6 concentration in the FMT group was lower than that in the control group [4.15 (1.91-5.71) ng/L vs 6.82 (2.40-8.91) ng/L, P=0.040], and the IL-10 concentration in the FMT group was higher than that in the control group [12.11 (5.69-20.36) ng/L vs 7.51 (4.10-9.58) ng/L, P=0.024]. Islet-derived protein 3α (REG3α) was significantly increased in patients with GI-aGVHD, and the REG3α level in the FMT group was lower than that in the control group after treatment [30.70 (10.50-105.00) μg/L vs 74.35 (33.50-139.50) μg/L, P=0.021]. Conclusion: FMT is a safe and effective method for the treatment of steroid-refractory GI-aGVHD by restoring intestinal microbiota diversity, regulating inflammatory cytokines, and upregulating Treg cells.

Effect of Jiawei Shenqi Yixin Prescription on Cardiovascular Risk Factors in Patients with Heart Failure with Preserved Ejection Fraction and Insulin Resistance / 中国实验方剂学杂志

ObjectiveTo observe the effect of Jiawei Shenqi Yixin prescription on cardiovascular risk factors in the patients with heart failure with preserved ejection fraction and insulin resistance. MethodFrom January 2021 to January 2022， a total of 82 patients with heart failure with preserved ejection fraction were enrolled in the ward of the First Affiliated Hospital of Heilongjiang University of Chinese Medicine. The patients were randomly assigned into two groups （ 41 cases） and received the same basic treatment. The observation group was additionally treated with Jiawei Shenqi Yixin prescription for 8 weeks. The clinical efficacy， traditional Chinese medicine （TCM） efficacy， cardiac function indexes ［NT-probrain natriuretic peptide （NT-proBNP） and 6-min walking test （6MWT）］， echocardiographic parameters ［left atrial volume index （LAVI）， left ventricular mass index （LVMI）， peak early diastolic to peak late diastolic mitral flow velocity （E/A） ratio］， insulin resistance-related indexes ［fasting insulin （FINS）， fasting plasma glucose （FPG）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， homeostatic model assessment of insulin resistance （HOMA-IR）， triglyceride-glucose index （TYG）， and triglyceride to high-density lipoprotein cholesterol （TG/HDL-C） ratio］， inflammatory cytokines ［tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， adiponectin （ADP）， and C-reactive protein （CRP）］， vascular endothelial function indicators ［nitric oxide （NO）， endothelial nitric oxide synthase （eNOS）， inducible nitric oxide synthase （iNOS）， and endothelin-1 （ET-1）］， and the safety of treatment were determined. In addition， Pearson correlation analysis was performed to analyze the correlations of insulin resistance， inflammatory cytokines， and vascular endothelial factors with the mitigation of heart failure. ResultIn terms of clinical efficacy， the therapy of the observation group was significantly effective in 26 patients， effective in 12 patients， ineffective in 3 patients， with the total effective rate of 92.68%， the therapy of the control group was significantly effective in 14 patients， effective in 12 patients， and ineffective in 15 patients， with the total effective rate of 63.41%. The clinical total effective rate of the observation group was higher than that of the control group （χ2=11.6， P<0.05）. In terms of TCM efficacy， the therapy of the observation group was significantly effective in 26 patients， effective in 11 patients， and ineffective in 4 patients， with the total effective rate of 90.24%； the therapy of the control group was significantly effective in 9 patients， effective in 13 patients， and ineffective in 19 patients， with the total effective rate of 53.66%. The TCM total effective rate of the observation group was higher than that of the control group （χ2=8.19， P<0.05）. Compared with those before treatment， the levels of NT-proBNP， LAVI， LVMI， FPG， FINS， HOMA-IR， TYG， TG/HDL-C， TNF-α， IL-6， CRP， ET-1， and iNOS in two groups declined after treatment （P<0.05）， while the levels of 6MWT， E/A， ADP， NO， and eNOS elevated （P<0.05）. After treatment， the observation group had lower levels of NT-proBNP， LAVI， LVMI， FPG， FINS， HOMA-IR， TYG， TG/HDL-C， TNF-α， CRP， and ET-1 （P<0.05） and higher levels of 6MWT， E/A， ADP， and NO than the control group （P<0.05）. In addition， the increase in 6MWT after treatment was positively correlated with the increase in NO and the decrease in ET-1. The decrease in LVMI after treatment was positively correlated with the increase in NO and the decrease in FINS. The increase in left ventricular ejection fraction after treatment was positively correlated with the decreases in TNF-α and TYG （P<0.05）. Adverse reactions were observed in neither group. ConclusionJiawei Shenqi Yixin prescription can significantly mitigate the symptoms， reduce inflammation， and improve vascular endothelial function in the patients with heart failure with preserved ejection fraction and insulin resistance， being safe without causing adverse reactions.

Alcohol Extract of Angelicae Pubescentis Radix Treats Gouty Arthritis Induced by MSU via NLRP3/ASC/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the efficacy and mechanism of the alcohol extract DH50 of Angelicae Pubescentis Radix in treating gouty arthritis induced by monosodium urate （MSU） crystals in vivo and in vitro. MethodFifty male SD rats were randomly assigned into five groups （n=10）： a normal group， a model group， a dexamethasone （DXMS， 0.07 mg·kg-1） group， and low- （DH50-D， 9 mg·kg-1） and high-dose （DH50-G， 18 mg·kg-1） DH50 groups. The rats in the normal group and model group were administrated with the same amount of pure water. On day 5， the gouty arthritis model was established by injecting MSU into the right ankle joint of rats. The toe volume and joint inflammation index were measured 4， 8， 24， and 48 h after modeling. The pathological changes of the synovial tissue were detected by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， and IL-6 in the synovial tissue. Western blot was employed to measure the protein levels of NOD-like receptor protein 3 （NLRP3）， cysteine-aspartic protease-1 （Caspase-1）， apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain （ASC）， IL-1β， and cyclooxygenase-2 （COX-2） in the synovial tissue. Furthermore， the cell inflammation model was established with RAW264.7 cells stimulated with MSU （75 mg·L-1）. The cell experiments were carried out with 6 groups： a normal group， a model group， a positive drug （DXMS， 100 μmol·L-1） group， and low- （DH50-D， 25 mg·L-1）， medium- （DH50-Z， 50 mg·L-1）， and high-dose （DH50-G， 100 mg·L-1） DH50 groups. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the cell viability， ELISA to determine the content of TNF-α in the supernatant of cell culture， and Western blot to determine the protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2. ResultCompared with the normal group， the rat model group showed increased toe swelling degree and joint inflammatory index （P<0.01）， serious infiltration of the synovium， elevated levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， ASC， IL-1β， and COX-2 （P<0.05， P<0.01）. Compared with the rat model group， low- and high-dose DH50 mitigated the toe swelling degree， decreased the joint inflammatory index， alleviated the inflammatory infiltration， lowered the levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and down-regulated the expression of related proteins （P<0.05， P<0.01）. Compared with the normal group， the cell model group showed elevated level of TNF-α in the supernatant （P<0.01） and up-regulated protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2 （P<0.05）. Compared with the model group， low， medium， and high doses of DH50 lowered the level of TNF-α in the supernatant of cell culture in a dose-dependent manner and down-regulated the expression of related proteins （P<0.05， P<0.01）. ConclusionDH50 can mitigate gouty arthritis both in vitro and in vivo by inhibiting the activation of NLRP3 inflammasomes and the production of inflammatory cytokines.

Improvement of inflammatory cytokines level in peripheral blood, anxiety and depressive symptoms in patients with pulpitis by root canal therapy / 口腔疾病防治

Objective@# To investigate the effect of root canal therapy (RCT) on inflammatory cytokines level in peripheral blood, anxiety, and depression in patients with pulpitis.@*Methods @#A total of 155 patients with pulpitis admitted to the Stomatology Hospital of the Fourth Military Medical University from June 2021 to June 2022 were treated with root canal therapy. Another 155 persons who received health examinations during the same period were selected as the control group. The Generalized Anxiety Disorder-7 (GAD-7) scores and the Patient Health Questionnaire-9 (PHQ-9) scores of the two groups were compared. The GAD-7, PHQ-9 and pain scores of the test group before treatment and 3 and 6 weeks after treatment were compared. Pain was assessed with a visual analog scale (VAS). Inflammatory cytokine [interleukin-8 (IL-8), interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α), c-reactive protein (CRP)] levels in the test group before treatment and 3 and 6 weeks after treatment were compared. @*Results @#The GAD-7 and PHQ-9 scores in the test group were higher than those in the control group before treatment (P<0.05). The GAD-7 and PHQ-9 scores in the test group at 3 and 6 weeks after treatment were lower than those before treatment (P<0.05); there was no significant difference between the GAD-7 and PHQ-9 scores at 3 and 6 weeks after treatment(P>0.05). The pain scores of the experimental group at 3 and 6 weeks after treatment were lower than those before treatment (P<0.05), and the pain scores 6 weeks after treatment were lower than those at 3 weeks after treatment (P<0.05). The levels of IL-8, IL-1β, TNF-α and CRP in the peripheral blood of the experimental group were lower 3 and 6 weeks after treatment than before (P<0.05), and the levels of IL-8 and IL-1β in the peripheral blood at 6 weeks after treatment were significantly lower than at 3 weeks after treatment (P<0.05). The levels of TNF-α and CRP in the peripheral blood at 6 weeks after treatment were not significantly different from those at 3 weeks after treatment (P>0.05).@*Conclusion @#The peripheral blood of patients with pulpitis has a high level of inflammatory cytokines, and the patients suffer from obvious anxiety and depression. Root canal therapy can relieve their anxiety and depression by reducing their level of inflammatory cytokines in peripheral blood.