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2022 ◽  
Vol 12 (4) ◽  
pp. 739-746
Zhihong Qiu ◽  
Li Yan ◽  
Juan Xu ◽  
Xiaojun Qian

Objective: The aim of our research was to evaluate Nrf2 in COPD treatment and relative mechanism by vivo study. Materials: The mice were divided into Normal, Model and CCL16 groups. Measuring Pathology and goblet cell number by HE or AB/PAS staining; Evaluating apoptosis cell number by TUNEL assay; using flow separation to analysis inflammatory cells in difference groups; MAPK and NF-κB(p65) protein expression were evaluated by IHC assay in tissues; Total protein concentration of MUC5AC, Nrf2, Bax and Bcl-2 were evaluated by WB assay. Results: Compared with Normal group, the pathology was deteriorate and goblet cell number were significantly up-regulation in Model group, apoptosis goblet cell number were significantly depressed (P < 0.001), lympbocyte rate and hypertrophic rate were significantly down-regulation and Eosinophils rate, Macrophage rate and Neutrophils rate were significantly up-regulation (P < 0.001, respectively) in Model group. By IHC assay, MAPK and NF-κB(p65) proteins expression significantly increased (P < 0.001, respectively) in Model group; by WB assay, MUC5AC and Bcl-2 protein expression were significantly up-regulation and Nrf2 and Bax proteins expression were significantly down-regulation (P < 0.001, respectively) in Model group. Nrf2 supplement, the COPD were significantly improved with relative inflammatory cells rates significantly improving and relative proteins improving. Conclusion: Nrf2 could improve COPD by inducing goblet cell apoptosis increasing via regulation MAPK/NF-κB(p65) pathway in vivo study.

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Xiang Zheng ◽  
Tao-Xiu Xiong ◽  
Ke Zhang ◽  
Fu-Chen Zhou ◽  
Hui-Ying Wang ◽  

Aim and Objective. To study the effect of Dendrobium officinale ultrafine powder (DOFP) on the intestinal mucosal barrier in rats with ulcerative colitis (UC) induced by dextran sulfate sodium (DSS). Materials and Methods. After intragastric administration of DOFP for 3 weeks, the rat UC model was made by the administration of 4% oral DSS solution for one week, and the drug was given at the same time. During the experiment, the disease activity index (DAI) score of the rats was regularly computed. At the end of the experiment, the blood routine indexes of rats were obtained. The histopathological changes in the colon were monitored by hematoxylin-eosin (H&E) and PAS staining and observation of ultrastructural changes in the colon by transmission electron microscope. Occludin expression in the colon was monitored by Western blot, the expression of claudin-1 and ZO-1 in the colon was detected by immunofluorescence, and the expression of TNF-α, IL-6, and IL-1β in the colon was detected by immunohistochemistry. Results. The results firstly indicated that DOFP could significantly alleviate the signs and symptoms of the DSS-induced rats UC model, which manifested as improvement of body weight loss, increase of colon length, and improvement of the symptoms of diarrhea and hematochezia. Then, results from histopathology, blood routine examination, and transmission electron microscope analysis further implied that DOFP could dramatically reduce inflammatory cell infiltration and restore intestinal epithelial barrier integrity. In addition, the experiments of Western Blot analysis, immunofluorescence, and PAS staining also further confirmed that DOFP could markedly increase related protein expressions of the intestinal barrier and mucus barrier, as the expression of occludin, claudin-1, and ZO-1 in the colon significantly decreased. The experiments of immunohistochemistry confirmed that DOFP could markedly decrease protein expression levels of inflammatory cytokines TNF-α, IL-6, and IL-1β. Conclusion. DOFP notably alleviated inflammatory lesions, repaired the colon mucosa damage by promoting the expression of tight junction proteins occludin, claudin-1, and ZO-1 and inhibiting the release of inflammatory factors TNF-α, IL-6, and IL-1β, and finally achieved the purpose of treating UC.

2021 ◽  
Vol 13 ◽  
Xinhua Zhan ◽  
Marisa Hakoupian ◽  
Lee-Way Jin ◽  
Frank R. Sharp

Corpora amylacea (CA) increase in number and size with aging. Their origins and functions remain unknown. Previously, we found that Alzheimer's disease (AD) brains have more CA in the periventricular white matter (PVWM) compared to aging controls. In addition, CA is associated with neurodegeneration as indicated by colocalization of degraded myelin basic protein (dMBP) with periodic acid-Schiff (PAS), a CA marker. We also found that bacterial lipopolysaccharide is present in aging brains, with more LPS in AD compared with controls. Periodic acid-Schiff staining is used to identify CA by virtue of their high polysaccharide content. Despite the growing knowledge of CA as a contributor to AD pathology, the molecules that contribute to the polysaccharides in CA are not known. Notably, lipopolysaccharides (LPS) are important cell-surface polysaccharides found in all Gram-negative bacteria. However, it is unknown whether PAS could detect LPS, whether the LPS found in aging brains contribute to the polysaccharide found in CA, and whether LPS associate with myelin injury. In this study, we found that aging brains had a myelin deficit zone (MDZ) adjacent to the ventricles in PVWM. The MDZ contained vesicles, most of which were CA. LPS and dMBP levels were higher in AD than in control brains. LPS was colocalized with dMBP in the vesicles/CA, linking white matter injury with a bacterial pro-inflammatory molecule. The vesicles also contained oxidized fibers, C-reactive protein, NG2, and GALC, markers of oligodendrocyte precursor cells (OPCs) and oligodendrocyte cells (OLs), respectively. The vesicles/CA were surrounded by dense astrocyte processes in control and AD brains. LPS was co-localized with CA by double staining of PAS with LPS in aging brains. The relationship of LPS with PAS staining was confirmed by PAS staining of purified LPS on nitrocellulose membranes. These findings reveal that LPS is one of the polysaccharides found in CA which can be stained with PAS. In addition, vesicles/CA are associated with oxidized and damaged myelin. The LPS in these vesicles/CA may have contributed to this oxidative myelin damage and may have contributed to oxidative stress to OPCs and OLs which could impair the ability to repair damaged myelin in AD and control brains.

2021 ◽  
Peipei Su ◽  
Dongwei Liu ◽  
Sijie Zhou ◽  
Hang Chen ◽  
Xianming Wu ◽  

Abstract BackgroundLncRNA AK044604 (Risa), Sirt1 and GSK3β are autophagy related genes that can play important roles in diabetic nephropathy (DN). In this study, we sought to explore the effect of Risa on Sirt1/GSK3β-induced podocyte injury.MethodsTransgenic db/db mice were fed and injected with Risa inhibition of adeno-associated virus by tail vein injection, as well as intraperitoneally injected with LiCl. Blood, urine, kidney tissue samples, and clinical data were collected at different time points. Immortalized mouse podocyte cells (MPCs) were cultured and treated with Risa inhibition of lentivirus, EX-527, and LiCl. MPCs were collected under different stimulations. The effects of Risa on podocyte autophagy were examined by qRT-PCR, Western blot analysis, transmission electron microscope, PAS staining, and immunofluorescence staining. ResultsRisa and activated GSK3β were overexpressed, but Sirt1 decreased in Renal tissues of DN mice and high-glucose-treated MPCs and correlated with poor prognosis. Risa overexpression attenuated Sirt1-mediated downstream autophagy levels and aggravated the injury of podocytes by inhibiting the expression of Sirt1. In contrast, Risa inhibition enhanced Sirt1-induced autophagy and attenuated podocyte injury, but this effect could be abrogated by EX-527, suggesting that Risa overexpression aggravated podocyte injury by decreasing autophagy. ConclusionsRisa inhibits autophagy by regulating the Sirt1/GSK3β axis and thereby aggravates podocyte injury in DN. Risa may serve as a therapeutic target for the treatment of DN.

2021 ◽  
Vol 16 (1) ◽  
Weifeng Wu ◽  
Yifan Wang ◽  
Haidi Li ◽  
Haiyong Chen ◽  
Jiangang Shen

Abstract Background Buyang Huanwu Decoction (BHD) is a classical Chinese Medicine formula empirically used for diabetic nephropathy (DN). However, its therapeutic efficacies and the underlying mechanisms remain obscure. In our study, we aim to evaluate the renoprotective effect of BHD on a streptozotocin (STZ)-induced diabetic nephropathy mouse model and explore the potential underlying mechanism in mouse mesangial cells (MCs) treated with high glucose in vitro, followed by screening the active compounds in BHD. Methods Mice were received 50 mg/kg streptozotocin (STZ) or citrate buffer intraperitoneally for 5 consecutive days. BHD was intragastrically administrated for 12 weeks starting from week 4 after the diabetes induction. The quality control and quantitative analysis of BHD were studied by high-performance liquid chromatography (HPLC). Renal function was evaluated by urinary albumin excretion (UAE) using ELISA. The mesangial matrix expansion and renal fibrosis were measured using periodic acid-schiff (PAS) staining and Masson Trichrome staining. Mouse mesangial cells (MCs) were employed to study molecular mechanisms. Results We found that the impaired renal function in diabetic nephropathy was significantly restored by BHD, as indicated by the decreased UAE without affecting the blood glucose level. Consistently, BHD markedly alleviated STZ-induced diabetic glomerulosclerosis and tubulointerstitial injury as shown by PAS staining, accompanied by a reduction of renal inflammation and fibrosis. Mechanistically, BHD inhibited the activation of TGF-β1/Smad3 and NF-κB signaling in diabetic nephropathy while suppressing Arkadia expression and restoring renal Smad7. We further found that calycosin-7-glucoside (CG) was one of the active compounds from BHD, which significantly suppressed high glucose-induced inflammation and fibrosis by inhibiting TGF-β1/Smad3 and NF-κB signaling pathways in mesangial cells. Conclusion BHD could attenuate renal fibrosis and inflammation in STZ-induced diabetic kidneys via inhibiting TGF-β1/Smad3 and NF-κB signaling while suppressing the Arkadia and restoring renal Smad7. CG could be one of the active compounds in BHD to suppress renal inflammation and fibrosis in diabetic nephropathy.

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1438
Souheil Zayet ◽  
Pierre Isnard ◽  
Jacinta Bustamante ◽  
David Boutboul ◽  
Sarra Abroug ◽  

Whipple’s Disease is a rare systemic infectious disease caused by the ubiquitous actinomycetes Tropheryma whipplei (T. whipplei). We report herein a rare case of a cutaneous granulo matosis with hypercalcemia as an unusual presenting feature of Whipple’s disease. The diagnosis of the bacteria was obtained from skin and inguinal lymph node biopsy (16 rDNA PCR screening and histological examination using PAS staining). T. whipplei was also identified on saliva and stool specimens, using specific PCR and colonic biopsies. Treatment with hydroxychloroquine and doxycycline allowed a rapid resolution of symptoms with a complete recovery.

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S451-S452
Barbara Ximenes Braz ◽  
Chantal Soobhanath ◽  
Amelia B Thompson

Abstract Background Chronic peritonitis is an unusual manifestation of coccidiomycosis (CM) that is challenging to diagnose and manage due to its propensity for relapse. It is even more unusual to diagnose peritoneal CM in the pediatric population, with only two other cases reported in the literature. Methods We present the case of a previously healthy 5-year-old Filipino female in Florida who was diagnosed with peritoneal CM. After months of unintentional weight loss and worsening abdominal distention, she presented to medical care. Imaging revealed significant abdominal ascites and nodularities throughout the peritoneum. The peritoneal fluid demonstrated a lymphocytic pleocytosis and infectious workup was benign. CA125 levels were elevated, but peritoneal adenosine deaminase was within normal limits. A biopsy of the affected tissue revealed diffuse granulomas surrounding spherules that were positive on GMS and PAS staining, concerning for CM. Exposure history revealed that she was raised in California and moved to Florida one year prior to presentation. Complement fixation titers were significantly elevated at ≥ 1:512 and immunodiffusion titers were positive. A Coccidioides PCR was sent from the tissue to the Mayo clinic and was positive, and fungal cultures from the tissue grew C. immitis/posadasii. Immunologic workup was reassuring. She was started on oral Fluconazole with rapid resolution of her symptoms. Results Involvement of the peritoneum in CM is extremely rare. Abdominal distention due to ascites is the most common presentation, and the peritoneal fluid is typically exudative. Imaging may reveal peritoneal deposits which can mimic other infections and malignancy. Diagnosis can be based on histopathological demonstration of fungal structures, cultures, antibody testing, antigen detection and/or PCR. Treatment guidelines suggest azole therapy for nonmeningeal disseminated CM with at least 6–12 months of treatment for extrapulmonary coccidioidal soft tissue infection. Conclusion Peritoneal CM is an extremely uncommon condition and it is even more rare in the pediatric population, but should be considered in those in the appropriate clinical settings, particularly if they have history to suggest exposure to regions where this fungus is endemic. Disclosures All Authors: No reported disclosures

Zygote ◽  
2021 ◽  
pp. 1-8
Mohammad Veisi ◽  
Kamran Mansouri ◽  
Vahideh Assadollahi ◽  
Cyrus Jalili ◽  
Afshin Pirnia ◽  

Summary An in vitro spermatogonial stem cell (SSC) culture can serve as an effective technique to study spermatogenesis and treatment for male infertility. In this research, we compared the effect of a three-dimensional alginate hydrogel with Sertoli cells in a 3D culture and co-cultured Sertoli cells. After harvest of SSCs from neonatal mice testes, the SSCs were divided into two groups: SSCs on a 3D alginate hydrogel with Sertoli cells and a co-culture of SSCs with Sertoli cells for 1 month. The samples were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays and bromodeoxyuridine (BrdU) tracing, haematoxylin and eosin (H&E) and periodic acid–Schiff (PAS) staining after transplantation into an azoospermic testis mouse. The 3D group showed rapid cell proliferation and numerous colonies compared with the co-culture group. Molecular assessment showed significantly increased integrin alpha-6, integrin beta-1, Nanog, Plzf, Thy-1, Oct4 and Bcl2 expression levels in the 3D group and decreased expression levels of P53, Fas, and Bax. BrdU tracing, and H&E and PAS staining results indicated that the hydrogel alginate improved spermatogenesis after transplantation in vivo. This finding suggested that cultivation of SSCs on alginate hydrogel with Sertoli cells in a 3D culture can lead to efficient proliferation and maintenance of SSC stemness and enhance the efficiency of SSC transplantation.

Photonics ◽  
2021 ◽  
Vol 8 (9) ◽  
pp. 350
Nadia Dembskey ◽  
Heidi Abrahamse

(1) Background: Onychomycosis accounts for 50% of nail pathologies and is a therapeutic challenge due to an increase in resistance to antifungal agents. This study aimed to explore the effectiveness of 1064 nm diode laser irradiation for the treatment of Onychomycosis and establish a new set of laser parameters for effective and safe treatment; (2) Methods: An exploratory, single-blinded study was conducted on forty-five patients with toenail Onychomycosis. Digital images and nail clippings were taken for Periodic Acid-Schiff (PAS) staining and fungal microscopy and culture (MC&S). Group 1 received 5% topical Amorolfine lacquer to apply to affected nails. Group 2 received 1064 nm diode laser treatment at 10 mW/s, hallux 790 J/cm2 and lesser digits 390 J/cm2 (standard treatment). Group 3 received 1064 nm diode laser treatment at 10 mW/s, hallux 1 100 J/cm2 and lesser digits 500 J/cm2 (new treatment parameters). After laser treatment, nail temperatures were taken with a surface thermometer; (3) Results: PAS staining was more sensitive in identifying Onychomycosis (91.1%), compared to Fungal Microscopy (44.4%). Comparing treatment requirements over a period of 24 weeks, there was a statistical significance, p ≤ 0.01 (**), for standard laser treatment and, p ≤ 0.001 (***), for new laser parameter treatment, indicating treatment needed over time decreased. No adverse effects were noted with new laser therapy. An 86.7% visual improvement was noted in Group 3 after 24 weeks; (4) Conclusions: Phototherapy, or photo thermolysis, was the best treatment option for Onychomycosis. A new protocol for the standardization of laser irradiation with the possible inclusion into the Scoring Clinical Index for Onychomycosis treatment plan, was proposed.

2021 ◽  
Xiaoyan Meng ◽  
Wenjing Huang ◽  
Weiwei Mo ◽  
Tingting Shu ◽  
Haoqiang Yang ◽  

Abstract Background: ADAMTS-13 plays an important role in acute kidney injury (AKI), but the mechanism of cisplatin (CP) induced AKI remains unclear. Ferroptosis is increased in CP-induced AKI, and ADAMTS13 levels are associated with ferritin expression. In this article, we will explore the relationship between the three.Methods: After CP induction, mice were given 0.1 and 0.3nmol/kg ADAMTS-13, and then Scr and BUN were detected by the kits. The pathological changes of renal tissue were observed by staining with HE and PAS staining, and Western blot detected the expressions of KIM1 and NGAL in renal tissu. Perl's staining detected iron deposition in renal tissues, the kits detected iron levels, and western blot detected the expression of ferroptosis related proteins. Then the mechanism was further explored by adding ferroptosis inhibitors Ferrostatin 1 (Fer-1) and iron supplements Fe. The expression of Nrf2 pathway related proteins were detected by Western blot.Results: ADAMTS13 alleviated CP-induced ferroptosis in AKI mice with renal function impairment and tubular damage. Fer-1partially reversed CP-induced AKI, and Fe exacerbated this effect. ADAMTS13 alleviated CP-induced inflammatory response and oxidative stress in AKI mice, during which the Nrf2 signaling pathway was abnormal.Conclusion:ADAMTS-13-regulated Nrf2 signaling inhibits ferroptosis to ameliorate CP-induced AKI.

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