Assessment of the levels of fibrogenic genes in human corneal fibroblasts.

<p>(A) Cells were treated with the indicated concentration of NIC or COT. After 24 hours’ incubation, mRNA expression of αSMA and TGFβ1 was determined by qRT-PCR (B) Cells were treated with the indicated concentrations of NIC or COT along with 0.01N NaOH to induce a chemical injury. After 24 hours’ incubation, mRNA expression of αSMA and TGFβ1 was determined by qRT-PCR. (C) Protein levels of αSMA were determined by Western blotting in total cell lysates. Values presented are from two independent experiments and each sample was assayed in duplicate. Data are expressed as the mean ± SEM per group and analyzed by ANOVA. Experimental groups marked by different letters (a, b, or c) represent significant difference between groups at p<0.05.</p

Evaluation of fibrogenic gene expression in injured corneal tissue.

<p>(A) mRNA levels of the fibrogenic genes αSMA, TGFβ1, and Col1 were analyzed. (B) Corneal tissues 14 days after injury were stained with an anti-αSMA antibody. Pictures are shown at 200x magnification. The percentage of αSMA-positive area was evaluated in corneal tissues. (C) αSMA protein levels were determined by Western blotting in corneal tissues with or without NIC treatment 14 days after alkali burn injury. Values are expressed as the mean ± SEM. Data was analyzed by ANOVA. Experimental groups marked by different letters (a, b, or c) represent significant difference at p<0.05 compared to the control group on each day. N.S., not significant.</p

Alkali injury-induced corneal NV in corneas with or without NIC treatment.

<p>(A) Stereomicroscopic appearance of mouse eyes on days 7 and 14. The length of new blood vessels was determined. (B) Results of qRT-PCR to detect VEGF and MMP9 mRNA levels are shown. Data are expressed as the mean ± SEM per group and analyzed by ANOVA. Experimental groups marked by different letters (a, b, or c) represent significant difference at p<0.05 compared to the control group on each day. N.S., not significant.</p

Comparison of nAChR subunit expression.

<p>(A) A representative qRT-PCR analysis showing expression of the α1–10, β1–4, γ, δ, and ε nAChR subunits in corneal fibroblasts. Values presented are from three independent experiments and each sample was assayed in duplicate. (B) mRNA levels of α3, α7, and β1 nAChR subunits in corneal tissues 14 days after injury. Data are expressed as the mean ± SEM per group and analyzed by Two-tailed Student’s <i>t</i> test; *P < 0.05, **P < 0.01. N.S., not significant.</p

Determination of the intracellular signaling molecules involved in corneal fibroblast proliferation induced by treatment with NIC or COT.

<p>The specific inhibitors WO and RO were used to inhibit PI3K and PKC, respectively. (A and B) Cell proliferation was determined by the CCK-8 assay. Values presented are from three independent experiments. Data are expressed as the mean ± SEM per group and analyzed by Two-tailed Student’s <i>t</i> test; *P < 0.05, **P < 0.01.</p

Additional file 1 of TLR7 activation by miR-21 promotes renal fibrosis by activating the pro-inflammatory signaling pathway in tubule epithelial cells

Additional file 1: Supplementary Table 1. Information of Antibodiesused in Western blotting. SupplementaryTable 2. Primer sequences for qPC

sj-docx-1-npx-10.1177_1934578X231180709 - Supplemental material for Antiviral and Therapeutic Effects of a Mixture of <i>Boswellia serrata</i>, <i>Commiphora myrrha</i>, and Propolis for SARS-CoV-2

Supplemental material, sj-docx-1-npx-10.1177_1934578X231180709 for Antiviral and Therapeutic Effects of a Mixture of Boswellia serrata, Commiphora myrrha, and Propolis for SARS-CoV-2 by Myeon-Sik Yang, Yun-Sook Lim, Byungkwan Oh, Seok-Chan Park, Daram Yang, Soon B Hwang and Bumseok Kim in Natural Product Communications</p

Presentation_1_Dual TLR2/9 Recognition of Herpes Simplex Virus Infection Is Required for Recruitment and Activation of Monocytes and NK Cells and Restriction of Viral Dissemination to the Central Nervous System.PDF

<p>The importance of TLR2 and TLR9 in the recognition of infection with herpes simplex virus (HSV) and HSV-caused diseases has been described, but some discrepancies remain concerning the benefits of these responses. Moreover, the impact of TLR2/9 on innate and adaptive immune responses within relevant mucosal tissues has not been elucidated using natural mucosal infection model of HSV. Here, we demonstrate that dual TLR2/9 recognition is essential to provide resistance against mucosal infection with HSV via an intravaginal route. Dual TLR2/9 ablation resulted in the highly enhanced mortality with exacerbated symptoms of encephalitis compared with TLR2 or TLR9 deficiency alone, coinciding with highly increased viral load in central nervous system tissues. TLR2 appeared to play a minor role in providing resistance against mucosal infection with HSV, since TLR2-ablated mice showed higher survival rate compared with TLR9-ablated mice. Also, the high mortality in dual TLR2/9-ablated mice was closely associated with the reduction in early monocyte and NK cell infiltration in the vaginal tract (VT), which was likely to correlate with low expression of cytokines and CCR2 ligands (CCL2 and CCL7). More interestingly, our data revealed that dual TLR2/9 recognition of HSV infection plays an important role in the functional maturation of TNF-α and iNOS-producing dendritic cells (Tip-DCs) from monocytes as well as NK cell activation in VT. TLR2/9-dependent maturation of Tip-DCs from monocytes appeared to specifically present cognate Ag, which effectively provided functional effector CD4⁺ and CD8⁺ T cells specific for HSV Ag in VT and its draining lymph nodes. TLR2/9 expressed in monocytes was likely to directly facilitate Tip-DC-like features after HSV infection. Also, dual TLR2/9 recognition of HSV infection directly activated NK cells without the aid of dendritic cells through activation of p38 MAPK pathway. Taken together, these results indicate that dual TLR2/9 recognition plays a critical role in providing resistance against mucosal infection with HSV, which may involve a direct regulation of Tip-DCs and NK cells in VT. Therefore, our data provide a more detailed understanding of TLR2/9 role in conferring antiviral immunity within relevant mucosal tissues after mucosal infection with HSV.</p

Data_Sheet_1_Blood parameters and pathological lesions in pigs experimentally infected with Vietnam's first isolated African swine fever virus.docx

African swine fever virus (ASFV) is a notable virus and one of the most serious global threats to the pig industry. Improving awareness about host–virus interactions could facilitate the understanding of the disease pathogenesis. Therefore, we investigated changes in blood parameters, viral loads, and pathological changes in ASFV-inoculated pigs according to the time of death after the onset of viremia. For the analyses, the ASFV-infected pigs (n = 10) were divided into two groups (five pigs/group) according to their time of death after the onset of viremia. The blood cell count dynamics and serum biochemistry profiles were similar between the groups; however, viral load distribution was different. A comparison of the histopathological changes and immunohistochemistry results between the two groups indicated that the lymphoid system, particularly the spleen, was more damaged in the early stage of the disease than in the last stage. Additionally, the virus-induced lesions in other organs (liver and kidney) were more severe in the late stage than in the early stage. Our findings provide invaluable information on the characteristics of blood parameters and pathological lesions in pigs infected with the Asia-epidemic ASFV strain and the course of ASF, targeting internal organs in pigs. Overall, this study characterizes the host-pathogen interaction in ASFV infection, offering insight for the establishment of ASF control strategies.</p