Expression of SMARCA2 and SMARCA4 in gastric adenocarcinoma and construction of a nomogram prognostic model.

BACKGROUND: Aberrant expression of SWI/SNF complex subunits is closely associated with tumorigenesis. The clinicopathological and prognostic significance of altered SMARCA2 and SMARCA4 subunits has not been well evaluated in gastric adenocarcinoma. METHODS: We collected 1271 postoperative cases of gastric adenocarcinoma and then constructed tissue microarrays (TMA), from which we obtained the immunohistochemistry expression of SMARCA2 and SMARCA4. Next, we screened the variables related to the loss of SMARCA2 and SMARCA4 by univariate correlation analysis and multivariate logistic regression analysis. Then, we identified the variables related to prognosis by univariate and multivariate Cox regression analysis. Finally, we constructed a nomogram prognostic model and evaluated it. RESULTS: The loss of SMARCA2 and SMARCA4 occurred in 236 (18.57%) and 86 (6.77%) cases, respectively, including 26 cases of co-loss. After multivariate logistic regression, variables independently associated with SMARCA2 loss were T stage, differentiation status, WHO histological classification, and EBER. Variables independently associated with SMARCA4 loss were differentiation status, WHO histological classification, PD-L1, and MMR. Survival analysis revealed that the SMARCA2 and SMARCA4 lost groups showed worse survival than the corresponding present groups (P = 0.032 and P = 0.0048, respectively). Univariate and multivariate Cox analyses identified independent prognostic factors, including age, T stage, N stage, M stage, SMARCA2, and chemotherapy. CONCLUSION: The loss of SMARCA2 and SMARCA4 correlated with poor differentiation, leading to a worse prognosis. SMARCA2, as an independent prognostic factor, combined with other clinicopathological variables, established a novel nomogram prognostic model, which outperformed the AJCC TNM model.

Role of NLR family pyrin domain-containing 3 inflammasome in the activation of pancreatic stellate cells.

NLRP3 inflammasome activation plays an important role in the development of pancreatic fibrosis. However, it is unclear whether the activation of the NLRP3 inflammasome is directly involved in the activation of Pancreatic stellate cells (PSCs). The aim of this study was to investigate the role and mechanism of the NLRP3 inflammasome in the activation of PSCs. In vivo, a rat model of chronic pancreatitis (CP) was induced by intravenous injection of dibutyltin dichloride (DBTC). In vitro, rat primary PSCs were isolated from pancreatic tissues and incubated with the NLRP3 inflammasome activator LPS, the NLRP3 inhibitor MCC950, or NLRP3 siRNA. The results showed that the expression of NLRP3, pro-Caspase-1, Caspase-1 and IL-18 was increased in the rat model of CP and during PSCs activation. LPS increased the protein levels of NLRP3, ASC, Caspase-1, IL-1ß and IL-18 accompanied by the upregulation of α-SMA, Col I and FN expression. Moreover, MCC950 or NLPR3 siRNA decreased the expression of α-SMA, Col I, FN, TGF-ß1 and p-Smad3. Furthermore, MCC950 reversed the LPS-induced upregulation of α-SMA, FN and Col Ⅰ expression in PSCs. This study revealed that the NLRP3 inflammasome is directly involved in the activation of PSCs in vivo and in vitro. Inhibiting NLRP3 suppresses the activation of PSCs through the TGF-ß1/Smad3 pathway.

Clinicopathological and prognostic significance of SWI/SNF complex subunits in undifferentiated gastric carcinoma.

BACKGROUND: The switch/sucrose nonfermentable (SWI/SNF) complex is an evolutionarily conserved chromatin remodeling complex that displays dysfunction in many tumors, especially undifferentiated carcinoma. Cancer stem cells (CSC), a special type of undifferentiated cancer cells with stem cell-like properties, play an essential role in tumor cell proliferation, invasion, and metastasis. In undifferentiated gastric carcinomas, the association of SWI/SNF complexes with clinicopathological features, CSC phenotype, and the prognosis is not fully understood. METHODS: We collected a cohort of 21 patients with undifferentiated/dedifferentiated gastric carcinoma. We next performed immunohistochemistry staining for the five subunits of the SWI/SNF complex (ARID1A, ARID1B, SMARCA2, SMARCA4, and SMARCB1), and four mismatch repair proteins (MLH1, PMS2, MSH2, and MSH6), as well as other markers such as p53, PD-L1, and cancer stem cell (CSC) markers (SOX2, SALL4). Then, we investigated the correlation of SWI/SNF complex subunits with clinicopathological characters and performed prognostic analysis. RESULTS: We observed SMARCA2 loss in 12 cases (57.14%), followed by ARID1A (5 cases, 23.81%) and SMARCA4 (3 cases, 14.29%). Fourteen cases (66.67%) lost any one of the SWI/SNF complex subunits, including 3 cases with SMARCA2 and ARID1A co-loss, and 3 cases with SMARCA2 and SMARCA4 co-loss. Correlation analysis revealed that the CSC phenotype occurred more frequently in the SWI/SNF complex deficient group (P = 0.0158). Survival analysis revealed that SWI/WNF complex deficiency, undifferentiated status, CSC phenotype, and the loss of SMARCA2 and SMARCA4 resulted in worse survival. Univariate and multivariate Cox regression analyses screened out three independent factors associated with worse prognosis: undifferentiated status, SWI/SNF complex deficiency, and lymph node metastasis. CONCLUSIONS: The SWI/SNF complex deficiency was more likely to result in a CSC phenotype and worse survival and was an independent prognostic factor in undifferentiated/dedifferentiated gastric carcinoma.

SMARCA2 deficiency in NSCLC: a clinicopathologic and immunohistochemical analysis of a large series from a single institution.

BACKGROUND: SMARCA2 (SWI/SNF Related, Matrix Associated, Actin Dependent Regulator of Chromatin, Subfamily A, Member 2) is an important ATPase catalytic subunit in the switch-sucrose nonfermenting (SWI/SNF) complex. However, its relationship with the pathological features of NSCLC and its prognosis remain unclear. METHODS: We retrospectively reviewed 2390 patients with surgically resected NSCLC, constructed tissue microarrays (TMAs) and performed immunohistochemical assays. We analyzed the correlation of SAMRCA2 with clinicopathological features and evaluated its prognostic value. RESULTS: Among 2390 NSCLC cases, the negative expression ratios of SAMRCA2, SMARCA4, ARID1A, ARID1B and INI1 were 9.3%, 1.8%, 1.2%, 0.4% and 0%, respectively. In NSCLC, male sex, T3 and T4 stage, moderate and poor differentiation, tumor ≥ 2 cm, Ki67 ≥ 15%, SOX-2 negative expression, middle lobe lesion and adenocarcinoma were relative risk factors affecting SMARCA2-negative expression. In lung adenocarcinomas, high-grade nuclei, histological morphology of acinar and papillary, solid and micropapillary and TTF-1-negative expression were relative risk factors affecting SMARCA2-negative expression. Kaplan-Meier survival analysis showed that the OS was shorter in the SMARCA2-negative group. Multivariate survival analysis revealed that SMARCA2-negative expression was an independent factor correlated with a poor prognosis in NSCLC. CONCLUSION: In conclusion, SMARCA2-negative expression is an independent predictor of a poor outcome of NSCLC and is a potential target for NSCLC treatment.

Construction of a survival prediction model for high-and low -grade UTUC after tumor resection based on "SEER database": a multicenter study.

BACKGROUND: There are differences in survival between high-and low-grade Upper Tract Urothelial Carcinoma (UTUC). Our study aimed to develop a nomogram to predict overall survival (OS) of patients with high- and low-grade UTUC after tumor resection, and to explore the difference between high- and low-grade patients. METHODS: Patients confirmed to have UTUC between 2004 and 2015 were selected from the Surveillance, Epidemiology and End Results (SEER) database. The UTUCs were identified and classified as high- and low-grade, and 1-, 3- and 5-year nomograms were established. The nomogram was then validated using the Chinese multicenter dataset (patients diagnosed in Shandong, China between January 2010 and October 2020). RESULTS: In the high-grade UTUC patients, nine important factors related to survival after tumor resection were identified to construct nomogram. The C index of training dataset was 0.740 (95% confidence interval [CI]: 0.727-0.754), showing good calibration. The C index of internal validation dataset was 0.729(95% CI:0.707-0.750). On the other hand, Two independent predictors were identified to construct nomogram of low-grade UTUC. The C index was 0.714 (95% CI: 0.671-0.758) for the training set,0.731(95% CI:0.670-0.791) for the internal validation dataset. Encouragingly, the nomogram was clinically useful and had a good discriminative ability to identify patients at high risk. CONCLUSION: We constructed a nomogram and a corresponding risk classification system predicting the OS of patients with an initial diagnosis of high-and low-grade UTUC.

Inhibitory Effects of the Extracts of Juglans sigillata Green Husks on the Proliferation, Migration and Survival of KYSE150 and EC9706 Human Esophageal Cancer Cell Lines.

This study evaluated the antitumor activity of the extracts of green husks of Juglans sigillata Dode on esophageal cancer. KYSE150 EC9706 cells were treated with different concentrations of six components of the extracts of J. sigillata green husks. Cell viability was measured by MTT. Cell migration and cell invasion were measured by wound-healing assays and transwell assays, respectively. Cell apoptosis and cycle were measured by flow cytometry. The expression of cell migration, cell cycle and cell apoptosis regulatory proteins was analyzed by Western blotting. Only the three constituents, including EtOH extractives, EtOAc soluble fraction and gallic acid (GA), exhibited inhibitory effects on the cell viability, migration and invasion by decreasing MMP2 and MMP9 expression (all P < 0.05). Flow cytometry revealed that these three constituents also induced cell apoptosis by increasing Bax and cleaved caspase-3 but decreasing Bcl-2 in KYSE150 and EC9706 cells. Furthermore, these constituents arrested the cell cycle at G0/G1 by downregulating the expression of Cyclin D1 but upregulating p53 and phospho-p53 expression in KYSE150 cells. In conclusion, the green husks of J. sigillata may act as a potential inhibitor on esophageal cancer growth. GA was the major single active constituent of the extracts.

Matrilin-2 prevents the TNFα-induced apoptosis of WB-F344 cells via suppressing JNK pathway.

Oval cells, a kind of hepatic progenitor cell quiescent at normal condition, activates to proliferate and differentiate into hepatocytes under severe and long-term liver injury, which usually raises severe inflammation. However, how oval cell survives in the inflammatory milieu interne is still unclear. Tumor necrosis factor α (TNFα), mimicking inflammatory hepatic milieu interne, was used to treat oval cell line, WB-F344, to test the protective function of matrilin-2. In this study, our data suggested that matrilin-2 prevented TNFα-induced apoptosis in WB-F344 cells via inhibiting ASK1/MKK7/JNK pathway. In conclusion, we determined that matrilin-2 plays the key role in maintaining the survival of oval cell and guarantees its proliferation under various injury factors.

Matrilin-2 regulates proliferation, apoptosis and cell cycle during radiation-induced injury in HPAEpiC cell.

Radiation pulmonary injury is related to the accumulation of extracellular matrix proteins in the alveolar interstitial space. Matrilin-2 as a component of extracellular filamentous networks, present higher level in the lung tissue from irradiated mice and irradiated pulmonary epithelial cell line, HPAEpiC cells. Knockdown of endogenous matrilin-2 prevents the apoptosis of HPAEpiC cell induced by the irradiation injury. Consistently, over-expression of matrilin-2 reduced the proliferation and induced apoptosis of HPAEpiC cells. Matrilin-2 promotes the expression of p21 via increasing the transcriptional activity of p53, by which induces the G1 phase arresting in HPAEpiC cells. In summary, matrilin-2, increased by irradiation, reduced the proliferation and induces apoptosis of pulmonary epithelial cells via p53/p21 pathway.

High glucose-induced Matrilin-2 expression in mouse mesangial cells was mediated by transforming growth factor beta 1 (TGF-ß1).

This study aimed at evaluating the effect of high glucose on the expression of extracellular matrix (ECM) protein Matrilin-2 and the mechanism underlying this effect by using a mouse mesangial cell line. Mouse mesangial cells (MMCs) were cultured in media containing normal (5 mM d-glucose) or high concentrations of glucose (30 mM d-glucose). The expression of Matrilin-2 was assessed by either RT-PCR or western blot. Additionally, transforming growth factor beta 1 (TGF-ß1) inhibitors and TGF-ß1 were used to determine whether glucose-regulated Matrilin-2 expression was mediated by the TGF-ß1/Smad3 signaling pathway. Our data demonstrated that Matrilin-2 expression was markedly induced by high glucose and TGF-ß1. High glucose-induced Matrilin-2 expression was inhibited by TGF-ß1/Smad3 inhibitors, indicating that Matrilin-2 was markedly induced by high glucose and this induction was mediated by the TGF-ß1/Smad3 pathway. Taken together, our results showed that high-glucose-induced Matrilin-2 expression that was mediated by the TGF-ß1/Smad3 signaling pathway might play a role in Diabetic nephropathy (DN) pathogenesis and our finding provided a potential diagnostic and/or therapeutic target for DN.

Overexpression of RRM2 in gastric cancer cell promotes their invasiveness via AKT/NF-κB signaling pathway.

The ribonucleotide reductase M2 subunit (RRM2) plays an active role in tumor progression and is frequently overexpressed in cancer. It plays a significant role in the regulation of cell invasiveness, cell migration and tumor metastasis. Elevated RRM2 expression has been reported to be associated with poor prognosis of gastric cancer. However, the molecular mechanisms of RRM2 in gastric cancer cells remain elusive. In our study, we found that RRM2 highly expressed in gastric cancer cells BGC823. RRM2 stimulation dose-dependently enhanced the invasion and migration of BGC823 cells. Furthermore, we found that the expressions of MMP-2 and MMP-9 in BGC823 cells were significantly increased after RRM2 stimulation. In addition, RRM2 time-dependently induced activation of AKT, IKBα, and NF-κB. These effects of RRM2 were prevented by AKT selective inhibitor GSK690693 as well as NF-κB selective inhibitor BAY117082. In conclusion, our findings establish a signaling role for RRM2 in gastric cancer cells and identify that the RRM2/AKT/NF-κB signaling pathway is essential for tumor invasiveness in gastric cancer cells. Thus, our data may provide knowledge for using RRM2 as a novel target for effective diagnosis and treatment of gastric cancer.

Resistance in Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) to new chemistry insecticides.

The control of rice leaffolder, Cnaphalocrocis medinalis (Guenée), depended mainly on the insecticide application in China for a long time, and the resistance development impacted the effects of insecticide application. In this study, 13 conventional and new chemistry insecticides were assayed for the toxicities to the larvae of rice leaffolder collected from Nanning, Changsha, and Nanjing, China, with rice seedling dip method during 2011-2013. Among the tested chemicals, macrolide insecticides spinetoram, spinosad, abamectin, and emanectin benzoate have the highest toxicities, whereas monosultap and Bt have the least toxicities to this insect. Comparing with the baseline data established in 2010, the susceptibilities of rice leaffolder to chlorantraniliprole, metaflumizone, and tebufenozide are declining simultaneously and gradually in the three regions from 2011 to 2013, and C. medinalis are becoming resistance to chlorantraniliprole, metaflumizone, and tebufenozide. The synchronous decreases of susceptibility in three geographic populations were not observed for macrolide insecticides, indoxacarb, chlorpyrifos, monosultap, and Bt. The synchronous insecticide susceptibility declines in field populations of the migratory insect collected from different areas indicated resistance evolution, and the sequence application patterns of different insecticides should be scheduled to delay the further development of resistance along the migratory pathway of the rice leaffolder in China.

Antifungal agents against Aspergillus niger for rearing rice leaffolder larvae (Lepidoptera: Pyralidae) on artificial diet.

Mold contamination is an important issue in insect mass rearing. Frequently used antifungal agents such as sorbic acid and methylparaben have negative impact on many lepidopteran larvae, which might be one of the reasons for the difficulty in rearing rice leaffolder, Cnaphalocrocis medinalis (Güenée). In this study, 19 antifungal agents, including 7 food preservatives, 6 antifungal drugs, and 6 agricultural fungicides, were screened for their inhibitory activities on Aspergillus niger in diets. The results demonstrated that most of the tested chemicals are unsuitable as mold inhibitors in the diets of the rice leaffolder, and the rice leaffolder neonate is sensitive to sorbic acid and methylparaben. These two mold inhibitors at commonly used concentrations were shown to impact the survival of rice leaffolder larvae fed on artificial diets. Among the tested mold inhibitors, natamycin was the safest for the rice leaffolder larvae. Much higher larva survival was observed for the larvae fed on diets containing natamycin as an antifungal agent (59 and 72% at 200 and 400 ppm, respectively). Two agricultural fungicides, tebuconazole and azoxystrobin, are also potent as mold inhibitors when used in insect diets. The mixed use of natamycin and sorbic acid, or methylparaben, and the mixed use of sorbic acid and azoxystrobin resulted in significantly higher larva survival than sorbic acid + methylparaben. Natamycin + azoxystrobin and sorbic acid + tebuconazole resulted in larva survival similar to that of sorbic acid + methylparaben. The ternary combination of natamycin, sorbic acid, and methylparaben was the best combination for the rearing of rice leaffolder.

Macro/Microfracture evolution and instability behaviors of high-temperature granite under water-cooling subjected to Brazilian splitting test using the DIC technique.

To investigate the evolution and stability characteristics of granite thermal damage, a series of Brazilian splitting tests is conducted on high-temperature granite samples using digital image correlation (DIC) technology. The results show that the Brazilian tensile strength and P-wave velocity exhibit a clear decline beyond a temperature threshold of 450~600°C, with a linear relationship between them. The presence of micro-cracks alters the stress transfer path, disrupting the stress balance on the Brazilian disc and leading to complex fracture patterns. At temperatures below 450°C, high strain areas and the development of micro-cracks occur at both the upper and lower loading ends of the granite Brazilian disc. However, these phenomena are only observed at the upper loading end when the temperature exceeds 450°C. Thermal cracks also cause changes in the internal structure of rock samples, and temperature variations can affect both the P-wave velocity and tensile strength. In terms of the relationship between P-wave velocity and Brazilian tensile strength (BTS) of high-temperature granite under water cooling, the negative exponential function model proposed in this study fits the experimental data very well.

Shape memory and antibacterial chitosan-based cryogel with hemostasis and skin wound repair.

Massive damage to the skin can lead to heavy bleeding and potential wound infection. Therefore, the preparation of low-cost wound dressings that meet these requirements by simple methods has a good application prospect. In the study, a shape memory cryogel prepared at low temperatures by mixing chitosan (CS) and citric acid (CA). Silver nanoparticles (Ag NPs) introduced into the cryogel through the reduction of Ag+ with tannic acid (TA) as a reducing agent. The CS/CA/Ag cryogel has good mechanical properties and interconnected macroporous structures. The results of hemostasis tests show that CS/CA/Ag cryogel can absorb a large amount of blood and promote blood cell adhesion compared with commercial gelatin sponges and gauze. Meanwhile, CS/CA/Ag cryogel has a good antibacterial ability against S. aureus and E. coli. Furthermore, CS/CA/Ag cryogel significantly promotes wound healing in the full-thickness wound model infected with S. aureus. In conclusion, the cryogel prepared by the simple method has great advantages in rapid hemostasis and promoting wound healing.

The Effect of Conductive Polyaniline on the Anti-Fouling and Electromagnetic Properties of Polydimethylsiloxane Coatings.

In this paper, four conductive polyaniline powders doped in hydrochloric acid, sulfuric acid, phosphoric acid, and sulfonic acid were selected and blended with polydimethylsiloxane to prepare coatings with an electromagnetic absorption effect and fouling desorption effect, respectively. A UV spectrophotometer was used to evaluate the settling rate of the powders. Fourier transform infrared spectrometry, laser confocal microscopy, and scanning electron microscopy were used to observe the morphology and structure of the powder and the coating. The interface properties of the coatings were characterized using a contact angle measurement, the mechanical properties of the coatings using a tensile test, and the electromagnetic properties of the powders and microwave absorption properties of the coatings using vector network analyzers. Meanwhile, the antifouling performance of the coatings was evaluated via the marine bacteria adhesion test and benthic diatom adhesion test, and the effect of conductive polyaniline on the antifouling performance of the coating was analyzed. The results show that adding polyaniline reduced the surface energy of the coating and increased the roughness, mechanical properties and anti-fouling properties of the coating. Moreover, adding appropriate polyaniline powder can enhance the electromagnetic wave loss of the coating. The followings values were recorded for a hydrochloric-acid-doped polyaniline coating: lowest surface energy of 17.17 mJ/m2, maximum fracture strength of 0.95 MPa, maximum elongation of 155%, maximum bandwidth of 3.81 GHz, and peak of reflection loss of -23.15 dB. The bacterial detachment rate of the polydimethylsiloxane (PDMS) samples was only 30.37%. The bacterial adhesion rates of the composite coating containing hydrochloric-acid-doped polyaniline were 4.95% and 2.72% after rinsing and washing, respectively, and the desorption rate was 45.35%. The chlorophyll concentration values were 0.0057 mg/L and 0.0028 mg/L, respectively, and the desorption rate was 54.62%.

Injectable and photothermal antibacterial bacterial cellulose cryogel for rapid hemostasis and repair of irregular and deep skin wounds.

For irregular and deep skin wounds, it's difficult for wound dressing to reach the injured site to achieve rapid hemostasis and provide wound protection. Bacterial cellulose (BC) has high strength and natural three-dimensional pore structure, which endows it shape recovery ability after absorbing blood when injected to the wound. Therefore, in the study, an injectable aldehyde bacterial cellulose/polydopamine (DBC/PDA) photothermal cryogel was prepared by oxidation polymerization method for hemostasis and repair of irregular and deep skin wounds. BC was oxidized by NaIO4 to form DBC and dopamine (DA) was introduced into DBC by reacting with the aldehyde group in DBC through Schiff base reaction. Under oxidation effect of NaIO4 and with freezing condition, water crystallization led to local aggregation of DA and DBC, and at the same time DA was oxidized to PDA and polymerized with DA on DBC. After the melting process, the porous cryogel was obtained. The introduction of PDA enhances the photothermal properties of DBC/PDA cryogel. DBC/PDA cryogel can kill most bacteria and provide wound protection under near-infrared light. In vitro and in vivo hemostatic tests show that the DBC/PDA cryogel can quickly absorb blood and stop bleeding. Combined with its good injectable, DBC/PDA cryogel can provide rapid hemostatic and protection in the face of irregular and deep skin wounds.

Anti-marine biofouling adhesion performance and mechanism of PDMS fouling-release coating containing PS-PEG hydrogel.

Polystyrene microspheres compounded with polyethylene glycol-based hydrogel (PS-PEG)/polydimethylsiloxane (PDMS) coatings were prepared using the physical blending method. The chemical structure, surface and interface properties, interlayer adhesion, and tensile properties were tested in this paper. Furthermore, the antifouling performance was evaluated through bovine serum albumin fluorescent protein adsorption testing, marine bacteria adhesion testing, and benthic diatom adhesion testing. The results showed that the coating performance was best when 20 wt% PS-PEG hydrogel was added. Its surface energy was only 19.21 mJ/m2, the maximum breaking strength was 1.24 MPa, the maximum elongation rate was 675 %, the elastic modulus was 2.53 MPa, and the anti-stripping rate was 100 %. In addition, the coating with added 20 wt% PS-PEG hydrogel bacterial adherence rate was 5.36 % and 2.45 % after rinsing and washing, respectively, and the removal rate was 54.29 %. In the benthic diatom adhesion test, the chlorophyll concentration a-value was only 0.0017 mg/L after washing with added 20 wt% hydrogel, and the protein desorption rate was 84.19 % higher than PDMS in the fluorescent protein adsorption test. This coating has the 'low adhesion' and 'desorption' characteristics in the three growth stages of biofouling. Meanwhile, the low surface energy of the silicone is stable, and the hydrogel also dynamically migrates to the surface to gradually form a hydration layer, both are synergistic. When 20 wt% PS-PEG hydrogel was added, the coating demonstrated excellent antifouling performance due to its high hydration layer, low surface energy, high elasticity, and high interlayer adhesion. This research is expected to contribute to the practical applications of hydrogel coatings in marine antifouling.

S1P/S1PR2 promote pancreatic stellate cell activation and pancreatic fibrosis in chronic pancreatitis by regulating autophagy and the NLRP3 inflammasome.

Sphingosine-1-phosphate (S1P) is a bioactive lipid molecule that governs various functions by embedding its receptor, S1PR, in different cells. Chronic pancreatitis (CP) is characterized by pancreatic fibrosis via activation of pancreatic stellate cells (PSCs). However, the effect of S1P on CP and PSC activation is still unknown. Here, we conducted a series of experiments to explore the effect of S1P on a CP rat model and primary cultured PSCs. In vivo, CP was induced by intravenous injection of dibutyltin dichloride. S1P was administered at a dosage of 200 µg/kg body weight per day by intraperitoneal injection. After 4 weeks, serum, plasma and pancreas samples were collected for molecular analysis and histological detection. In vitro, PSCs were isolated and cultured for treatment with different doses of S1P. 3MA and MCC950 were used to determine the effect of S1P on PSC activation by regulating autophagy and the NLRP3 inflammasome. JTE013 and Si-S1PR2 were applied to verify that the functions of S1P were realized by combining with S1PR2. Cells were collected for RT‒PCR, western blotting and immunofluorescence. The results showed that S1P was increased in the plasma and pancreatic tissue of CP rats. When S1P was administered to CP rats, the function and histomorphology of the pancreas were severely impaired. In addition, S1P promoted PSC activation, heightened autophagy and enhanced the NLRP3 inflammasome in vivo and in vitro. Moreover, S1PR2 mediated the effect of S1P on PSC activation by regulating autophagy and the NLRP3 inflammasome sequentially. In conclusion, S1P binding to S1PR2 promoted PSC activation and pancreatic fibrosis in CP by regulating autophagy and the NLRP3 inflammasome. These findings provide a theoretical basis for targeting S1P/S1PR2 to treat pancreatic fibrosis and further suggest that considering the role of autophagy and the NLRP3 inflammasome may help with the treatment pancreatic fibrosis.

Prognostic and immune infiltration significance of ARID1A in TCGA molecular subtypes of gastric adenocarcinoma.

BACKGROUND: AT-rich interaction domain 1A (ARID1A) is an essential subunit of the switch/sucrose non-fermentable chromatin remodeling complex and is considered to be a tumor suppressor. The Cancer Genome Atlas (TCGA) molecular classification has deepened our understanding of gastric cancer at the molecular level. This study explored the significance of ARID1A expression in TCGA subtypes of gastric adenocarcinoma. METHODS: We collected 1248 postoperative patients with gastric adenocarcinoma, constructed tissue microarrays, performed immunohistochemistry for ARID1A, and obtained correlations between ARID1A and clinicopathological variables. We then carried out the prognostic analysis of ARID1A in TCGA subtypes. Finally, we screened patients by random sampling and propensity score matching method and performed multiplex immunofluorescence to explore the effects of ARID1A on CD4, CD8, and PD-L1 expression in TCGA subtypes. RESULTS: Seven variables independently associated with ARID1A were screened out: mismatch repair proteins, PD-L1, T stage, differentiation status, p53, E-cadherin, and EBER. The independent prognostic variables in the genomically stable (GS) subtype were N stage, M stage, T stage, chemotherapy, size, and ARID1A. PD-L1 expression was higher in the ARID1A negative group than in the ARID1A positive group in all TCGA subgroups. CD4 showed higher expression in the ARID1A negative group in most subtypes, while CD8 did not show the difference in most subtypes. When ARID1A was negative, PD-L1 expression was positively correlated with CD4/CD8 expression; while when ARID1A was positive, this correlation disappeared. CONCLUSIONS: The negative expression of ARID1A occurred more frequently in the Epstein-Barr virus and microsatellite instability subtypes and was an independent adverse prognostic factor in the GS subtype. In the TCGA subtypes, ARID1A negative expression caused increased CD4 and PD-L1 expression, whereas CD8 expression appeared independent of ARID1A. The expression of CD4/CD8 induced by ARID1A negativity was accompanied by an increase in PD-L1 expression.

Xuanfei Baidu Decoction suppresses complement overactivation and ameliorates IgG immune complex-induced acute lung injury by inhibiting JAK2/STAT3/SOCS3 and NF-κB signaling pathway.

BACKGROUND: The significant clinical efficacy of Xuanfei Baidu Decoction (XFBD) is proven in the treatment of patients with coronavirus disease 2019 (COVID-19) in China. However, the mechanisms of XFBD against acute lung injury (ALI) are still poorly understood. METHODS: In vivo, the mouse model of ALI was induced by IgG immune complexes (IgG-IC), and then XFBD (4g/kg, 8g/kg) were administered by gavage respectively. 24 h after inducing ALI, the lungs were collected for histological and molecular analysis. In vitro, alveolar macrophages inflammation models induced by IgG-IC were performed and treated with different dosage of XFBD-containing serum to investigate the protective role and molecular mechanisms of XFBD. RESULTS: The results revealed that XFBD mitigated lung injury and significantly downregulated the production of pro-inflammatory mediators in lung tissues and macrophages upon IgG-IC stimulation. Notably, XFBD attenuated C3a and C5a generation, inhibited the expression of C3aR and C5aR and suppressed the activation of JAK2/STAT3/SOCS3 and NF-κB signaling pathway in lung tissues and macrophages induced by IgG-IC. Moreover, in vitro experiments, we verified that Colivelin TFA (CAF, STAT3 activator) and C5a treatment markedly elevated the IgG-IC-triggered inflammatory responses in macrophages and XFBD weakened the effects of CAF or C5a. CONCLUSION: XFBD suppressed complement overactivation and ameliorated IgG immune complex-induced acute lung injury by inhibiting JAK2/STAT3/SOCS3 and NF-κB signaling pathway. These data contribute to understanding the mechanisms of XFBD in COVID-19 treatment.