[Correlations Between the Expression of MicroRNA-155 and Suppressor of Cytokine Signaling 1 in Colonic Mucosal Tissue and Disease Severity in Patients With Ulcerative Colitis].

Objective To explore the relationship between the expression levels of microRNA-155 (miR-155) and suppressor of cytokine signaling 1 (SOCS1) in the colonic mucosal tissue of patients with ulcerative colitis (UC) and the severity of the disease.Methods A total of 130 UC patients admitted to the Second Affiliated Hospital of Hebei North University from September 2021 to June 2023 were selected.According to the modified Mayo score system,the patients were assigned into an active stage group (n=85) and a remission stage group (n=45).According to the modified Truelove and Witts classification criteria,the UC patients at the active stage were assigned into a mild group (n=35),a moderate group (n=30),and a severe group (n=20).A total of 90 healthy individuals who underwent colonoscopy for physical examination or those who had normal colonoscopy results after single polypectomy and excluded other diseases were selected as the control group.The colonic mucosal tissues of UC patients with obvious lesions and the colonic mucosal tissue 20 cm away from the anus of the control group were collected.The levels of miR-155 and SOCS1 mRNA in tissues were determined by fluorescence quantitative PCR,and the expression of SOCS1 protein in tissues was determined by immunohistochemistry.The correlations of the levels of miR-155 and SOCS1 mRNA in the colonic mucosal tissue with the modified Mayo score of UC patients were analyzed.The values of the levels of miR-155 and SOCS1 mRNA in predicting the occurrence of severe illness in the UC patients at the active stage were evaluated.Results Compared with the control group and the remission stage group,the active stage group showed up-regulated expression level of miR-155,down-regulated level of SOCS1 mRNA,and decreased positive rate of SOCS1 protein in the colonic mucosal tissue (all P<0.001).The expression level of miR-155 and modified Mayo score in colonic mucosal tissues of UC patients at the active stage increased,while the mRNA level of SOCS1 was down-regulated as the disease evolved from being mild to severe (all P<0.001).The modified Mayo score was positively correlated with the miR-155 level and negative correlated with the mRNA level of SOCS1 in colonic mucosal tissues of UC patients (all P<0.001).The high miR-155 level (OR=2.762,95%CI=1.284-5.944,P=0.009),low mRNA level of SOCS1 (OR=2.617,95%CI=1.302-5.258,P=0.007),and modified Mayo score≥12 points (OR=3.232,95%CI=1.450-7.204,P=0.004) were all risk factors for severe disease in the UC patients at the active stage.The area under curve of miR-155 combined with SOCS1 mRNA in predicting severe illness in the UC patients at the active stage was 0.920.Conclusions The expression levels of miR-155 and SOCS1 mRNA were correlated with the disease severity in the UC patients at the active stage.The combination of the two indicators demonstrates good performance in predicting the occurrence of severe illness in UC patients at the active stage.

Downregulation of Nogo-B ameliorates cerebral ischemia/reperfusion injury in mice through regulating microglia polarization via TLR4/NF-kappaB pathway.

Many studies have shown a close association between Nogo-B and inflammation-related diseases. However, uncertainty does exist, regarding Nogo-B function in the pathological progression of cerebral ischemia/reperfusion (I/R) injury. Middle cerebral artery occlusion/reperfusion (MCAO/R) model was utilized in C57BL/6L mice to mimic ischemic stroke in vivo. Using oxygen-glucose deprivation and reoxygenation (ODG/R) model in microglia cells (BV-2) to establish cerebral I/R injury in vitro. Various methods, including Nogo-B siRNA transfection, mNSS and the rotarod test, TTC, HE and Nissl staining, immunofluorescence staining, immunohistochemistry, Western blot, ELISA, TUNEL and qRT-PCR were employed to probe into the effect of Nogo-B downregulation on cerebral I/R injury and the potential mechanisms. A small amount of Nogo-B expression (protein and mRNA) was observed in cortex and hippocampus before ischemia, then Nogo-B expression increased significantly on day 1, reaching the maximum on day 3, remaining stable on day 14 after I/R, and decreasing gradually after 21 days, but it still rose significantly compared with that observed preischemia. Nogo-B down-regulation could markedly reduce the neurological score and infarct volume, improve the histopathological changes and neuronal apoptosis, lower the number of CD86+/Iba1+ cells and the levels of IL-1ß, IL-6, and TNF-α, and raise the density of NeuN fluorescence, the number of CD206+/Iba1+ cells, and the level of IL-4, IL-10 and TGF-ß in brain of MCAO/R mice. Treatment with Nogo-B siRNA or TAK-242 in BV-2 cells could obviously decrease the CD86 fluorescence density and the mRNA expression of IL-1ß, IL-6 and TNF-α, increase CD206 fluorescence density and the mRNA expression of IL-10 after OGD/R injury. In addition, the expression of TLR4, p-IκBα and p-p65 proteins significantly increased in the brain after MCAO/R and BV-2 cells exposed to OGD/R. Treatment with Nogo-B siRNA or TAK-242 prominently reduced the expression of TLR4, p-IκBα and p-p65. Our findings suggest that the down-regulation of Nogo-B exerts protective effect on cerebral I/R injury by modulating the microglia polarization through inhibiting TLR4/NF-κB signaling pathway. Nogo-B may be a potential therapeutic target for ischemic stroke.

Anfibatide alleviates inflammation and apoptosis via inhibiting NF-kappaB/NLRP3 axis in ischemic stroke.

Recent evidence suggests that Nod-like receptor protein-3 (NLRP3) inflammasome is a key mediator of inflammatory response and can induce the activation of apoptosis signaling pathways in ischemic stroke. In this research, we assessed the effects of anfibatide (ANF) on inflammatory and apoptosis in cerebral ischemic injury and the potential mechanisms. Middle cerebral artery occlusion (MCAO) model was established on male Sprague-Dawley rats to induce cerebral ischemia/reperfusion (I/R) injury in vivo. Primary cortical neurons (PCN) cells were exposed to oxygen-glucose deprivation and reintroduction (OGD/R) to mimic cerebral I/R injury in vitro. The results showed that ANF markedly alleviated infarct volume, neurological deficit and neurobehavioral impairment in MCAO/R rats, enhanced cell viability and decreased LDH release in PCN after OGD/R. The number of TUNEL-positive cells, Bax, cleaved-caspase-3, p-IκBα, p-p65, NLRP3, ASC, cleaved caspase-1, IL-ß and IL-18 proteins expression were significantly upregulated in the cortex of MCAO/R rats and PCN exposed to OGD/R, NLRP3 and caspase-1 mRNA levels were also evidently elevated. Bcl-2 protein expression significantly decreased in the cortex of MCAO/R rats. Treatment with ANF obviously inhibited the expression of p-IκBα, p-p65, NLRP3, ASC, cleaved caspase-1, Bax and cleaved-caspase-3, promoted the expression of Bcl-2, then decreased the TUNEL-positive cell number and the level of inflammatory cytokines (IL-ß and IL-18) in cerebral ischemia reperfusion in vito and in vitro. Our findings suggest that ANF exerts effects of alleviating inflammation and apoptosis through inhibiting NF-kappaB/NLRP3 axis. ANF is a potential candidate for treating cerebral I/R injury.

Anfibatide Preserves Blood-Brain Barrier Integrity by Inhibiting TLR4/RhoA/ROCK Pathway After Cerebral Ischemia/Reperfusion Injury in Rat.

The disruption of the blood-brain barrier (BBB) and the consequent brain edema are major contributors to the pathogenesis of cerebral ischemia/reperfusion injury. RhoA is generally thought to play a crucial role in the process of BBB disruption and participate in the signaling pathways emanating from TLR4. However, it remains unverified the regulatory role of TLR4 in the RhoA/ROCK pathway in cerebral I/R injury and its effects on the BBB as well. The present study probes into the protective effect of ANF on the BBB after cerebral I/R injury and the possible mechanisms. Focal cerebral ischemia was induced by 120 min of transient middle cerebral artery occlusion (MCAO). ANF (1, 2, 4 µg/kg) was achieved by intravenous injection after 120 min of MCAO followed by 1, 24, 48, and 72 h reperfusion. Evans blue extravasation, brain water content, RhoA activity, and the expressions of TLR4, ROCK1/2, p-MLC2, MMP-2/9, ZO-1, occludin, and claudin-5 protein in rat brain were evaluated 72 h after reperfusion. ANF could significantly reduce the Evans blue extravasation and water content in the ipsilateral hemisphere and obviously increase the occludin, claudin-5, and ZO-1 expression after cerebral I/R injury. Furthermore, cerebral I/R injury induced apparently increased expression of TLR4, RhoA-GTP, ROCK1/2, p-MLC2, and MMMP-2/9, which, however, could be remarkably alleviated by ANF intervention. Taken together, the TLR4/RhoA/ROCK signaling pathway is implicated in BBB breakdown after cerebral I/R injury, and ANF preserves BBB integrity, probably via inhibiting the TLR4/RhoA/ROCK signaling pathway.

Therapeutic effects of Baizhu Huanglian Decoction on ulcerative colitis in rats / 第二军医大学学报

Objective To explore the therapeutic effects of Baizhu Huanglian Decoction on rats with ulcerative colitis (UC) induced by trinitrobenzene sulfonic acid (TNBS). Methods SD rats were used to make rat UC model by TNBS enema. A total of 74 rats were divided into 6 groups with half of each group being male. Model group had 14 rats, and Baizhu Huanglian Decoction low-, middle- and high-dose groups, control group and sulfasalazine group had 12 rats in each group. One rat in the model group and one in the sulfasalazine group were dead on the 2nd day of modeling. After modeling, the rats of Baizhu Huanglian Decoction low-, middle- and high-dose group were intragastrically given (2 times a day, each time with 2 mL) Baizhu Huanglian Decoction of 6.895, 13.790 and 27.580 g/kg, respectively. Sulfasalazine group was given sulfasalazine suspension by gavage 0.2 g/kg (2 times a day, each time with 2 mL) and model group was given the same amount of normal saline after modeling. Normal control group was always given normal saline. After 10 days of continuous administration, the body mass and fecal characteristics of the rats were observed. At the end of the administration, the colonic pathological changes of colon and the length of colorectal were measured. The colon pathology and colon mucosal injury index (CMDI) score were evaluated. The levels of nitric oxide (NO), inducible nitric oxide synthase (iNOS), myeloperoxidase (MPO) and interleukin-6 (IL-6) were measured by ELISA. Results Compared with the model group, the body mass of the rats were significantly increased on the 4th-10th day after administration in the Baizhu Huanliang Decoction low-, middle- and high-dose groups and sulfasalazine group (all P0.05), which had no significant difference versus normal control group. Fecal characteristic scores of the Baizhu Huanglian Decoction high-dose groups and the sulfasalazine group were lower than those of the model group and the Baizhu Huanglian Decoction middle- and low-dose groups, but the differences were not statistically significant. Compared with the model group, the colorectal lengths of the rats were significantly longer in Baizhu Huanglian Decoction middle- and high-dose groups and sulfasalzine group (all P0.05). The colon pathology scores were significantly lower in the Baizhu Huanglian Decoction low-, middle- and high-dose group, and sulfasalazine group versus the model group (all P0.05). CMDI scores of Baizhu Huanglian Decoction high-dose group and sulfasalazine group were significantly lower than those of the model group (both P0.05). The levels of NO, iNOS, MPO and IL-6 of the rat colonic tissues were significantly lower in Baizhu Huanliang Decoction low-, middle- and high-dose groups, and sulfasalazine group than those in the model group (all P0.05). Conclusion Baizhu Huanglian Decoction can improve the general condition and colonic lesions of rats with UC, decreasing the levels of NO, iNOS, MPO and IL-6 and reducing the inflammation.

Protective effect and mechanism of Qishen Yiqi Formula on mitochondrial structure and function of cardiomyocytes injured by H2O2 / 中草药

Objective: To establish a cell-based multi-parameter mitochondrial structure and functional evaluation system for hydrogen peroxide (H2O2) injury in cardiomyocytes and investigate the protective effect of Qishen Yiqi Formula on H9c2 cells and its mechanism. Methods: For the in vitro myocardial cell injury model, H9c2 cells were divided into control, model (H2O2), positive control (carbonyl cyanide-cyanomethoxy basic hydrazine, 5 μmol/L), and Qishen Yiqi Formula (0.2 mg/mL) groups, with three duplicates in each group, and were cultured in triplicate for 24 h with corresponding drugs, followed by 2 h H2O2 induction. H2O2 injury model was established with H9c2 cell line. Mitochondrial function and morphological texture were evaluated by Operetta high content imaging system. Mitochondrial respiration and bioenergetic metabolism states were measured by Seahorse Bioscience XF extracellular flux analyzer. Cardiomyocyte apoptosis were quantified by flow cytometry. Results: Qishen Yiqi Formula prevented the decrease of mitochondrial membrane potential and the increase of mitochondrial mass and improved mitochondrial morphological integrity. Functionally, Qishen Yiqi Formula increased basic oxygen consumption rate, ATP-linked oxygen consumption rate, maximum oxygen consumption, and the reserve capacity. It also reduced the apoptotic rate at early and end stage and increased the myocyte survival. Conclusion: Based on the structure and functional evaluation of the mitochondria, results indicated that the compound Qishen Yiqi Formula protected the H9c2 cells by improving the mitochondrial function and energy metabolism as well as reducing the apoptosis.

[Protective effect of Wuziyanzong Pills on rats with experimental oligoasthenospermia and its action mechanism].

OBJECTIVE: To investigate the protective effect of Wuziyanzong Pills (WYP) in the rat model of oligoasthenospermia (OAS) and its action mechanism. METHODS: Sixty male SD rats were equally randomized into six groups: normal control, OAS model, Shengjing Capsules (1.6 g per kg of the body weight), low-dose WYP (1 g per kg of the body weight), medium-dose WYP (2 g per kg of the body weight), and high-dose WYP (4 g per kg of the body weight). The OAS model was established by intragastric administration of Tripterygium glucoside at 30 mg per g per d for 6 weeks. From the 3rd week of modeling, the rats of the medication groups were treated intragastrically with corresponding drugs for 4 weeks. Then all the rats were sacrificed for measurement of the testicular and epididymal organ coefficients, examination of epididymal sperm quality and apoptosis, and detection of the openness of the sperm mitochondrial permeability transition pore (MPTP). Histopathological changes in the testis were observed by HE staining and the apoptosis of spermatogenic cells determined by Hochest staining. RESULTS: WYP obviously improved the organ coefficients of the testis and epididymis, increased sperm concentration, motility and viability, decreased the apoptosis of spermatogenic cells, and inhibited the abnormal openness of MPTP in the OAS model rats. HE staining showed that the number and levels of spermatogenic cells were significantly increased while Hochest staining manifested that the apoptosis of spermatogenic cells was remarkably inhibited in the seminiferous tubules of the testis in the WYP-treated rats. CONCLUSIONS: WYP can improve sperm quality and reduce the apoptosis of spermatogenic cells (including sperm) in OAS model rats, which may be related with its inhibitory effect on the abnormal openness of MPTP.

Effects of different time on ischemia/reperfusion injury in isolated rat hearts / 中国药学杂志

METHODS: Male Sprague Dawley rats which were used lor Langendoff isolated heart perfusion were divided into four groups; normal control group (n=6), 120 group (n=1), 130 group (n=8) and 140 group (n=8), these hearts were subjected to global ischemia for 20, 30 and 40 min respectively. Then coronary flow, heart rate, creatinine kinase and lactate dehydrogenase in effluent and the changes of cardiac function parameters were measured in different groups. Infarct and risk areas were measured by planimetry using Image/J software.

Intracoronary nitroprusside in the prevention of the no-reflow phenomenon in acute myocardial infarction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>No-reflow phenomenon during percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI) is a predictive factor of continuous myocardial ischemia, ventricular remodeling and cardiac dysfunction, which is closely associated with a worse prognosis. This study aimed to evaluate intracoronary nitroprusside in the prevention of the no-reflow phenomenon in AMI.</p><p><b>METHODS</b>Ninety-two consecutive patients with AMI, who underwent primary PCI within 12 hours of onset, were randomly assigned to 2 groups: intracoronary administration of nitroprusside (group A, n = 46), intracoronary administration of nitroglycerin (group B, n = 46). The angiographic results were observed. The real-time myocardial contrast echocardiography (RT-MCE), including contrast score index (CSI), wall motion score index (WMSI), transmural contrast defect length (CDL) and serious WM abnormal length (WML) were recorded at 24 hours and 1 week post-PCI. High sensitivity C-reactive protein (Hs-CRP) was examined by immune rate nephelometry. N-terminal prohormone brain natriuretic peptide (NT-proBNP) was tested with enzyme-linked immunosorbent assay. Patients were followed up for six months. Major adverse cardiac events (MACE) were recorded.</p><p><b>RESULTS</b>The incidence of final TIMI-3 flow in group A was much higher than that in Group B (P < 0.05), final corrected TIMI frame count (cTFC) in group A decreased significantly than that in group B (P < 0.01). The CSI, CDL/LV length, WMSI and WL/LV length in group A were significantly lower than that in group B (P < 0.01). Levels of Hs-CRP and NT-proBNP at 1 week post-PCI decreased significantly in group A than that in group B (P < 0.01). Patients were followed up for 6 months and the incidence of MACE in group A was significantly lower than that in group B (P < 0.05).</p><p><b>CONCLUSION</b>Intracoronary nitroprusside can improve myocardial microcirculation, leading to the decrease of the incidence of no-reflow phenomenon and better prognosis.</p>

Initial study on discrimination of oral microorganisms with the metabonomics technique / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility of employing metabonomics method in identification of oral pathogenic bacteria.</p><p><b>METHODS</b>The Streptococcus mutans ATCC25175 and Actinomyces viscosus ATCC15987 were respectively inoculated in same certain culture medium. The growth curves of the inoculated bacteria were drown by turbidimetry. The culture solutions in four different growth phases of the both bacteria were used to test with the 1H-Nuclear magnetic resonance (1H-NMR) spectroscopy respectively. The data of 1H-NMR spectroscopy results were analyzed by principal components analysis (PCA).</p><p><b>RESULTS</b>The PCA showed the obvious clustering phenomena and the points of two group data stayed differentially together by two clusters. Therefore, the NMR-based metabonomics profiles can discriminate the two different kind of bacteria.</p><p><b>CONCLUSION</b>The metabonomics can be expected to be a kind of promising useful method in quick discrimination of oral pathogenic bacteria.</p>