PDIA3 orchestrates effector T cell program by serving as a chaperone to facilitate the non-canonical nuclear import of STAT1 and PKM2.

Dysregulated T cell activation underpins the immunopathology of rheumatoid arthritis (RA), yet the machineries that orchestrate T cell effector program remain incompletely understood. Herein, we leveraged bulk and single-cell RNA sequencing data from RA patients and validated protein disulfide isomerase family A member 3 (PDIA3) as a potential therapeutic target. PDIA3 is remarkably upregulated in pathogenic CD4 T cells derived from RA patients and positively correlates with C-reactive protein level and disease activity score 28. Pharmacological inhibition or genetic ablation of PDIA3 alleviates RA-associated articular pathology and autoimmune responses. Mechanistically, T cell receptor signaling triggers intracellular calcium flux to activate NFAT1, a process that is further potentiated by Wnt5a under RA settings. Activated NFAT1 then directly binds to the Pdia3 promoter to enhance the expression of PDIA3, which complexes with STAT1 or PKM2 to facilitate their nuclear import for transcribing T helper 1 (Th1) and Th17 lineage-related genes, respectively. This non-canonical regulatory mechanism likely occurs under pathological conditions, as PDIA3 could only be highly induced following aberrant external stimuli. Together, our data support that targeting PDIA3 is a vital strategy to mitigate autoimmune diseases, such as RA, in clinical settings.

Convergent Regulation of Candida albicans Aft2 and Czf1 in Invasive and Opaque Filamentation.

Candida albicans is the most common fungal pathogen of mucosal infections and invasive diseases in immuno-compromised humans. The abilities of yeast-hyphal growth and white-opaque switching affect C. albicans physiology and virulence. Here, we showed that C. albicans Aft2 regulator was required for embedded filamentous growth and opaque cell-type formation. Under low-temperature matrix embedded conditions, Aft2 functioned downstream of Czf1-mediated pathway and was required for invasive filamentation. Moreover, deletion of AFT2 significantly reduced opaque cell-type formation under N-acetylglucosamine (GlcNAc) inducing conditions. Ectopic expression of CZF1 slightly increased the white-opaque switching frequency in the aft2Δ/Δ mutant, but did not completely restore to wild-type levels, suggesting that Czf1 at least partially bypassed the essential requirement for Aft2 in response to opaque-inducing cues. In addition, multiple environmental cues altered AFT2 mRNA and protein levels, such as low temperature, physical environment and GlcNAc. Although the absence of Czf1 or Efg1 also increased the expression level of AFT2 gene, deletion of CZF1 remarkably reduced the stability of Aft2 protein. Furthermore, C. albicans Aft2 physically interacted with Czf1 under all tested conditions, whereas the interaction between Aft2 and Efg1 was barely detectable under embedded conditions, supporting the hypothesis that Aft2, together with Czf1, contributed to activate filamentous growth by antagonizing Efg1-mediated repression under matrix-embedded conditions.

Effect of Methyl-CpG binding domain protein 2 (MBD2) on AMD-like lesions in ApoE-deficient mice.

The role of methyl-CpG binding domain protein 2 (MBD2) in an ApoE-deficient mouse model of age-related macular degeneration (AMD) was investigated. Eight-week-old Mbd2/ApoE double deficient (Mbd2(-/-) ApoE(-/-)) mice (n=12, 24 eyes, experimental group) and MBD2 (wt) ApoE(-/-) mice (n=12, 24 eyes, control group) were fed on Western-type diet for 4 months. The mice were sacrificed, and total serum cholesterol levels were analyzed and Bruch's membrane (BM) of the eyes was removed for ultrastructural observation by transmission electron microscopy. Moreover, intercellular adhesion molecule 1 (ICAM-1) immunoreactivities were evaluated by fluorescence microscopy in sections of the eyes in both groups for further understanding the function mechanism of MBD2. There was no significant difference in the total serum cholesterol levels between control group and experimental group (P>0.05). Transmission electron microscopy revealed that AMD-like lesions, various vacuoles accumulated on BM, notable outer collagenous layer deposits and dilated basal infoldings of retinal pigment epithelium (RPE) were seen in both groups, and the BM in control group was significantly thickened as compared with experimental group (P<0.05). Fluorescence micrographs exhibited the expression of ICAM-1 in choroid was higher in control group than in experimental group. We are led to conclude that MBD2 gene knockout may lead to accumulation of more deposits on the BM and influence the pathogenesis of AMD via triggering endothelial activation and inflammatory response in choroid, improving microcirculation, and reducing lipid deposition so as to inhibit the development of AMD-like lesions. Our study helps to provide a new therapeutic approach for the clinical treatment of AMD.

GSK2334470 attenuates high salt-exacerbated rheumatoid arthritis progression by restoring Th17/Treg homeostasis.

High salt (HS) consumption is a risk factor for multiple autoimmune disorders via disturbing immune homeostasis. Nevertheless, the exact mechanisms by which HS exacerbates rheumatoid arthritis (RA) pathogenesis remain poorly defined. Herein, we found that heightened phosphorylation of PDPK1 and SGK1 upon HS exposure attenuated FoxO1 expression to enhance the glycolytic capacity of CD4 T cells, resulting in strengthened Th17 but compromised Treg program. GSK2334470 (GSK), a dual PDPK1/SGK1 inhibitor, effectively mitigated the HS-induced enhancement in glycolytic capacity and the overproduction of IL-17A. Therefore, administration of GSK markedly alleviated HS-exacerbated RA progression in collagen-induced arthritis (CIA) model. Collectively, our data indicate that HS consumption subverts Th17/Treg homeostasis through the PDPK1-SGK1-FoxO1 signaling, while GSK could be a viable drug against RA progression in clinical settings.

PDIA3 defines a novel subset of adipose macrophages to exacerbate the development of obesity and metabolic disorders.

Adipose tissue macrophages (ATMs) play important roles in maintaining adipose tissue homeostasis and orchestrating metabolic inflammation. Given the extensive functional heterogeneity and phenotypic plasticity of ATMs, identification of the authentically pathogenic ATM subpopulation under obese setting is thus necessitated. Herein, we performed single-nucleus RNA sequencing (snRNA-seq) and unraveled a unique maladaptive ATM subpopulation defined as ATF4hiPDIA3hiACSL4hiCCL2hi inflammatory and metabolically activated macrophages (iMAMs), in which PDIA3 is required for the maintenance of their migratory and pro-inflammatory properties. Mechanistically, ATF4 serves as a metabolic stress sensor to transcribe PDIA3, which then imposes a redox control on RhoA activity and strengthens the pro-inflammatory and migratory properties of iMAMs through RhoA-YAP signaling. Administration of Pdia3 small interfering RNA (siRNA)-loaded liposomes effectively repressed adipose inflammation and high-fat diet (HFD)-induced obesity. Together, our data support that strategies aimed at targeting iMAMs by suppressing PDIA3 expression or activity could be a viable approach against obesity and metabolic disorders in clinical settings.

Aloperine Ameliorates IMQ-Induced Psoriasis by Attenuating Th17 Differentiation and Facilitating Their Conversion to Treg.

Aloperine is an anti-inflammatory compound isolated from the Chinese herb Sophora alopecuroides L. Previously, our group has reported that the generation of induced Treg was promoted by aloperine treatment in a mouse colitis model. However, the effect of aloperine on effector T cell subsets remains unclear. We therefore carefully examined the effect of aloperine on the differentiation of major subsets of T helper cells. Based on our results, psoriasis, a Th17 dominant skin disease, is selected to explore the potential therapeutic effect of aloperine in vivo. Herein, we demonstrated that topical application of aloperine suppressed epidermal proliferation, erythema, and infiltration of inflammatory cells in skin lesions. Mechanistic studies revealed that aloperine suppressed the differentiation of Th17 cells directly through inhibiting the phosphorylation of STAT3 or indirectly through impairing the secretion of Th17-promoting cytokines by dendritic cells. Moreover, aloperine enhanced the conversion of Th17 into Treg via altering the pSTAT3/pSTAT5 ratio. Collectively, our study supported that aloperine possesses the capacity to affect Th17 differentiation and modulates Th17/Treg balance, thereby alleviating imiquimod (IMQ)-induced psoriasis in mice.

Highly efficient photocontrolled targeted delivery of siRNA by a cyclodextrin-based supramolecular nanoassembly.

In this study, we have constructed a binary supramolecular nanoassembly composed of α-cyclodextrin-modified hyaluronic acid and an azobenzene-modified diphenylalanine derivative with a positively charged imidazole group. This nanoassembly can bind with siRNA through electrostatic interactions and efficiently delivered them into cancer cells and inhibited their growth.

[Effects of perfluorooctane sulfonate on learning and memory of rat pups].

OBJECTIVE: To study the effects of prenatal and postnatal perfluorooctane sulfonate (PFOS) exposure on spatial learning and memory, N-methyl-D-aspartate receptor 2B (NR2B) mRNA and protein level in frontal cortex and hippocampus of rat pups and to explore the mechanism of developmental neurotoxicity induced by PFOS. METHODS: Twenty-eight pregnant rats were randomly divided into three groups in proportion of 3:2:2, including control group (C), low dose group (L) and high dose group (H) by means of randomized number table, which respectively received 0, 7.2, 14.4 mg/kg PFOS feed from pregnancy day 0 to postnatal day (PND) 30 by free feedings. The animal models of prenatal and postnatal non-exposure (CC), prenatal exposure (LC and HC), postnatal exposure (CL and CH), and prenatal and postnatal exposure (LL and HH) to PFOS were established by cross-fostering method. The spatial learning and memory were measured by water maze experiment,the NR2B mRNA levels in frontal cortex of rat pups was determined with semi-quantitative RT-PCR, NR2B protein express in cerebral cortex (frontal and temporal cortex) and hippocampus (CA1, CA3, CA4 and DG regions) of rat pups was detected by immunohistochemistry. RESULTS: The escape latency of CL, CH, LL and HH groups pups in water maze experiment were (99.83 +/- 25.77) s, (111.30 +/- 17.82) s, (106.40 +/- 18.71) s, (107.70 +/- 16.85) s, and longer as compared with CC group [(54.90 +/- 26.69) s] (q value were 4.349, 4.773, 6.026 and 5.641, respectively, P <0.01). The number of errors of HH group rat pups entering dead end was (22.30 +/- 7.56) at the training day 4, and it was significantly higher than that of CC group (9.80 +/- 4.64) (q = 5.173, P < 0.01). The NR2B mRNA levels of frontal cortex of pups in HC group at PND1, and LC group, HC group and HH group at PND14 were (0.167 +/- 0.008), (0.364 +/- 0.035), (0.341 +/- 0.030) and (0.328 +/- 0.045) respectively,which were significantly lower than CC group (0.271 +/- 0.060) and (0.465 +/- 0.067) (q values were 3.547, 3.739, 4.597 and 5.006, respectively, P< 0.05 ). The results of immunohistochemistry indicated that NR2B protein express of the hippocampus CA1 region of pups in LC group was (0.091 +/- 0.005), and showed significant lower than CC group which was (0.123 +/- 0.009) at PND1 (q = 5.209, P <0.05). At PND14, the effect of PFOS extended to cerebral cortex and hippocampus regions. At PND28, the effects of PFOS were showed in hippocampus CA1, CA3 and temporal cortex regions. CONCLUSION: Prenatal and postnatal exposure to PFOS should result in the spatial learning and memory damage,and the mechanism might be possibly involved in the decrease of NR2B level in cerebral cortex and hippocampal formation regions.

[Comparative study on the acute toxicity of pulmonary caused by nanosized and microsized powders of silicon dioxide].

OBJECTIVE: To compare the acute pulmonary toxicities of nanosized and microsized silicon dioxide particles. METHODS: All 125 healthy male Wistar rats were divided into 25 groups randomly according to the weight. Experimental animals were exposed to microsized SiO2 at the doses of 100 mg/m3 (group A) and 300 mg/m3 (group B), and to the nanosized SiO2 at the same dose levels (group A' and B') by inhalation for 2 hours. Compositions in bronchoalveolar lavage fluids (BALF) and contents of hydroxyproline in blood sera and lung tissues were detected and then compared at 6, 12, 24, 48, 72 hours after administration. RESULTS: The total cellular score (TCS) in BALF of group A'[(55.00 +/- 8.30) x 10(4)/ ml] and B'[(52.50 +/- 9.02) x 10(4)/ml] at 6 hours were significantly higher than those in control groups [(34.88 +/- 12.53) x 10(4)/ml]; TCS in BALF of group A' [(55.00 +/- 8.30) x 10(4)/ml]at 6 hours and group A' [(39.75 +/- 12.08) x 10(4)/ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(32.38 +/- 13.07) x 10(4)/ml, (24.13 +/- 10.97) x 10(4)/ml) ]; total protein (TPr) in BALF of group A' [(0.34 +/- 0.09)g/L] and B' [(0.38 +/- 0.16) g/L] at 48 hours were significantly higher than those in isodose group of microsized SiO2 [(0.20 +/- 0.07) g/L, (0.21 +/- 0.05) g/L]. Lactate dehydrogenase (LDH) in BALF of group A' [(1.66 +/- 0.22) x 10(3) U/L] at 72 hours were significantly higher than those in isodose group of microsized SiO2 [(1.38 +/- 0.17) x 10(3) U/L]. Alkaline phosphatase (AKP) in BALF of group B' [(5.14 +/- 1.47) U/100 ml] at 6 hours and group B' [(5.86 +/- 2.41) U/100 ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(3.64 +/- 0.36) U/100 ml, (3.30 +/- 2.19) U/100 ml]. Hydroxyproline (HyP) in tissues of lung of group A' [(0.532 +/- 0.053) microg/mg, (0.484 +/- 0.046) microg/mg, (0.591 +/- 0.096) microg/mg, (0.551 +/- 0.084) microg/mg] at 6, 12, 48, 72 hours and group B' [(0.508 +/- 0.081) microg/mg, (0.565 +/- 0.053) microg/mg ] at 12, 72 hours were significantly higher than those in isodose group of microsized SiO2 [(0.345 +/- 0.074) microg/mg, (0.368 +/- 0.095) microg/mg, (0.431 +/- 0.036) microg/mg, (0.399 +/- 0.080) microg/mg, (0.396 +/- 0.039) microg/mg, (0.465 +/- 0.062) microg/mg]. CONCLUSION: Nanosized and microsized SiO2 should have some differences on acute pulmonary toxicities in our experiment condition.

Assessing the impact of cigarette smoking on ß-cell function and risk for type 2 diabetes in a non-diabetic Chinese cohort.

Although the impact of cigarette smoking on glucose homeostasis has been extensively studied, the results, however, are still not conclusive. We, therefore, conducted a cross-sectional analysis of a non-diabetic Chinese cohort collected by the China Health and Nutrition Survey (CHNS 2009) to comprehensively assess the relationship between smoking, Hemoglobin A1c, ß-cell function and insulin sensitivity. The cohort included a total of 5965 individuals (47.4% male) with a mean age of 49.23 years, and 4140 of which were non-smokers (69.4%), 834 were current light smokers (13.9%) and 991 were current heavy smokers (16.6%). Current smokers were predominantly males (93.6%) with a lower BMI (22.95 versus 23.42 kg/m2). HbA1c levels were dose-dependently increased with smoking exposure (5.39%, 5.42% and 5.45%, respectively, P = 0.007). Non-smokers were served as a referent, the adjusted ORs for type 2 diabetes were 1.12 (P = 0.256, light smokers) and 1.26 (P = 0.014, heavy smokers), indicating a positive relationship between cigarette smoking and incidence of diabetes. HOMA%B was decreased in a dose-responsive manner with cigarette smoking (4.80, 4.79 and 4.76, P = 0.036), suggesting an adverse effect of smoking on ß-cell function. Collectively, cigarette smoking is dose-dependently associated with decreased HOMA%B, and current smokers were clearly in a higher risk for diabetes as manifested by the elevated HbA1c.

[Effects of perfluorooctane sulfonate on Glu, PKC and PKA activities in mouse brain].

OBJECTIVE: To study the effects of perfluorooctane sulfonate (PFOS) on contents of glutamate and activity of protein kinase C (PKC) and A (PKA) and ultrastructure injury in the brain of male mice and to explore the mechanism of neurotoxicity and patho-alteration resulted from PFOS. METHODS: 44 male mice were randomly divided into four groups, who were respectively orally given 0, 5, 10, 20 mg/kg PFOS for 10 days. The Glu consents in the brain of the mice was measured with spectrophotometer and protein kinases activity were measured with non-radioactive assay of protein kinase and the changes of cerebral cortex ultrastructure were observed. RESULTS: Contents of Glu in 10 and 20 mg/kg groups were (1.57 +/- 0.11) and (1.62 +/- 0.16) mmol/g prot respectively,which was significantly increased compared with the corresponding controlled group [(1.45 +/- 0.13) mmol/g prot] (F = 39.59, P < 0.05). PKC activity in 5, 10 and 20 mg/kg BW groups were (29.05 +/- 2.89), (33.65 +/- 3.82) and (34.20 +/- 3.16) pmol x min(-1) x (mg prot)-1 respectively, which was significantly increased compared with the corresponding control group [(24.53 +/- 2.88) pmol x min(-1) x (mg prot)-1] (F = 7.75, P < 0.05). Compared with the corresponding control group, PKA in 5, 10 and 20 mg/kg BW groups increased by (24.12 +/- 3.86)%, (34.02 +/- 3.04)% and (33.42 +/- 3.71)% with a statistical significance (F = 26.27, P < 0.01). The exposed mice had cerebral cortex ultrastructure injury of cell nucleus envelope hollow. CONCLUSION: Exposure to PFOS increases Glu contents and activity of PKC and PKA in mouse brain and induce the cerebral cortex ultrastructural injury, a possible mechanism of the neurotoxicity caused by PFOS.

Sumoylation modulates oxidative stress relevant to the viability and functionality of pancreatic beta cells.

Sumoylation is an evolutionarily conserved regulatory mechanism to play an important role in various cellular processes through modulation of protein localization, stability and functionality. Recent studies including ours have consistently demonstrated that sumoylation provides protection for cells against oxidative stress. Given that pancreatic beta cells are a vulnerable target of oxidative stress, we thus in this minireview, updated the advancement of sumoylation in the regulation of ROS generation, and discussed its impact on several critical signaling pathways relevant to beta cells against oxidative stress and maintenance of functionality. Specifically, we bring together how sumoylation represses intracellular ROS formation, and protects beta cells against oxidative stress through regulating IκB/NFκB, JNK/c-Jun, and Maf/Nrf2 pathways. The tight implication of sumoylation in oxidative stress reflects that it could be an essential mechanism for beta cells to adapt to the detrimental cellular microenvironment.

[Screening and nitrogen transformation activity of ammonia-oxidizing molds during aerobic chicken manure composting].

In order to explicit the existence and nitrogen transformation activity of ammonia-oxidizing molds during aerobic chicken manure composting, the medium of ammonia-oxidizing molds was used to screen them, and correlation analysis between mycelium biomass and nitrogen transformation indexes was conducted, then high-effective ammonia-oxidizing strains were determined and added into aerobic compost. The results show that all tested strains are able to oxidize ammonium nitrogen to nitrite nitrogen and nitrate nitrogen, indicating that there are plenty of ammonia-oxidizing molds during aerobic chicken manure composting. Significantly positive correlations are detected between total contents of nitrite nitrogen and nitrate nitrogen, mycelium biomass and mycelium Kjeldahl nitrogen. When cultured in the medium of ammonia-oxidizing molds for 144 hours, two high-effective ammonia-oxidizing strains, named M25-22 (Penicillium sp.) and M40-4 (Aspergillus sp.), produce nitrate nitrogen by 1.1 x 10(-3) mg x mL(-1), 1.5 x 10(-3) mg x mL(-1), respectively, and decrease ammonium nitrogen by approximately 0.3 mg x mL(-1). Compared with the control treatment, the composting treatment with microbial agent M25-22 or M40-4 has lower ammonium nitrogen content, and higher contents of nitrate nitrogen and total nitrogen, indicating that they are of practical significance in reducing nitrogen loss during aerobic composting.

[Investigation of PFOS and PFOA pollution in snow in Shenyang, China].

The purpose of this study was to investigate the atmospheric perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) pollution in Shenyang by analyzing the concentration of the two compounds in snow. Snow samples were collected from both urban and suburban areas on Feb. 6 (n=36) and Feb. 25 (n=5) in 2006 in Shenyang, China. PFOS and PFOA were extracted from the snowmelt by solid phase extraction and then were analyzed by LC-MS-SIM. PFOS and PFOA were discernable in all the samples. In the snow samples collected on Feb. 6, 2006, the geometric average concentrations of PFOS and PFOA were 2.0 ng x L(-1) (range: 0.4-46.2 ng x L(-1)) and 3.6 ng x L(-1) (range: 1.6-22.4 ng x L(-1)), respectively. The 95% confidence intervals of PFOS and PFOA were 1.5-2.8 ng x L(-1) and 3.1-4.2 ng x L(-1), respectively. The concentration of PFOS and PFOA showed significant correlation at the center of Shenyang City (n=9), while the highest concentrations of PFOS and PFOA were detected at the same site located in the rural area. Geometric average concentrations of PFOS and PFOA were both 2.2 ng x L(-1) in the Feb. 25 snow samples. Concentrations of PFOS in two snow events did not show significant difference. Whereas PFOA concentration in Feb. 25 snow was higher than that in Feb. 6 snow. Results suggested the widespread PFOS and PFOA pollution in snow in Shenyang. There may be similar resources for the two compounds in regional area. Moreover, there may be continual import of PFOS to the atmosphere in Shenyang from specific resources, while PFOS and PFOA may exhibit different environmental behavior.