Fragment-Based Anti-inflammatory Agent Design and Target Identification: Discovery of AF-45 as an IRAK4 Inhibitor to Treat Ulcerative Colitis and Acute Lung Injury.

UC and ALI are inflammatory diseases with limited treatment in the clinic. Herein, fragment-based anti-inflammatory agent designs were carried out deriving from cyclohexylamine/cyclobutylamine and several fragments from anti-inflammatory agents in our lab. AF-45 (IC50 = 0.53/0.60 µM on IL-6/TNF-α in THP-1 macrophages) was identified as the optimal molecule using ELISA and MTT assays from the 33 synthesized compounds. Through mechanistic studies and a systematic target search process, AF-45 was found to block the NF-κB/MAPK pathway and target IRAK4, a promising target for inflammation and autoimmune diseases. The selectivity of AF-45 targeting IRAK4 was validated by comparing its effects on other kinase/nonkinase proteins. In vivo, AF-45 exhibited a good therapeutic effect on UC and ALI, and favorable PK proprieties. Since there are currently no clinical or preclinical trials for IRAK4 inhibitors to treat UC and ALI, AF-45 provides a new lead compound or candidate targeting IRAK4 for the treatment of these diseases.

Design, synthesis and bioactivity evaluation of 4-hydroxycoumarin derivatives as potential anti-inflammatory agents against acute lung injury and colitis.

Acute lung injury (ALI) and inflammatory bowel disease (IBD) are common inflammatory illnesses that seriously affect people's health. Herein, a series of 4-hydroxylcoumarin (4-HC) derivatives were designed and synthesized. The inhibitory effects of these compounds on LPS-induced interleukin-6 (IL-6) release from J774A.1 cells were then screened via ELISA assay, compound B8 showed 3 times more active than the lead compound 4-HC. The most active compound B8 had the IC50 values of 4.57 µM and 6.51 µM for IL-6 release on mouse cells J774A.1 and human cells THP-1, respectively. Furthermore, we also found that B8 could act on the MAPK pathway. Based on the target prediction results of computer virtual docking, kinase inhibitory assay was carried out, and it revealed that targeting IRAK1 was a key mechanism for B8 to exert anti-inflammatory activity. Moreover, B8 exerted a good therapeutic effect on the dextran sulfate sodium (DSS)-induced colitis model and liposaccharide (LPS)-induced ALI mouse models. The acute toxicity experiments indicated that high-dose B8 caused no adverse reactions in mice, confirming its safety in vivo. Additionally, the preliminary pharmacokinetic (PK) parameters of B8 in SD rats were also examined, revealing a bioavailability (F) of 28.72 %. In conclusion, B8 is a potential candidate of drug for the treatment of ALI and colitis.

Discovery of novel osthole derivatives exerting anti-inflammatory effect on DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice.

The modification based on natural products is a practical way to find anti-inflammatory drugs. In this study, 26 osthole derivatives were synthesized, and their anti-inflammatory properties were evaluated. The preliminary activity study revealed that most osthole derivatives could effectively inhibit inflammatory cytokines IL-6 secretion in LPS stimulated mouse macrophages J774A.1. Compound 7m exhibited the most effective anti-inflammatory activity (RAW264.7 IL-6 IC50: 4.57 µM, 32 times more active than osthole) in vitro with no significant influence on cell proliferation. Additionally, the mechanistic analysis demonstrated that compound 7m could block MAPK signal transduction by inhibiting the phosphorylation of JNK and p38, thereby inhibiting the release of inflammatory cytokines. Moreover, in vivo functional investigations revealed that 7m could substantially reduce DSS-induced ulcerative colitis and LPS-induced acute lung injury, with good therapeutic effects. The pharmacokinetics and acute toxicity experiments proved the safety and reliability of 7min vivo. Overall, Compound 7m could further be studied as potential anti-inflammatory candidate.

Discovery of the Diphenyl 6-Oxo-1,6-dihydropyridazine-3-carboxylate/carboxamide Analogue J27 for the Treatment of Acute Lung Injury and Sepsis by Targeting JNK2 and Inhibiting the JNK2-NF-κB/MAPK Pathway.

Acute lung injury (ALI) and sepsis are both serious and complex conditions associated with high mortality, yet there are no effective treatments. Herein, we designed and synthesized a series of diphenyl 6-oxo-1,6-dihydropyridazine-3-carboxylate/carboxamide analogues exhibiting anti-inflammatory activity. The optimal compound J27 decreased the release of TNF-α and IL-6 in mouse and human cells J774A.1 and THP-1 (IL-6 IC50 = 0.22 µM) through the NF-κB/MAPK pathway. J27 demonstrated remarkable protection against ALI and sepsis in vivo and exhibited good safety in subacute toxicity experiments. Pharmacokinetic study indicated that J27 had good bioavailability (30.74%). To our surprise, J27 could target JNK2 with a totally new molecular skeleton compared with the only few JNK2 inhibitors reported. Moreover, there is no report that JNK2 inhibitors could apply for ALI and sepsis. Therefore, this work provides a new lead structure for the study of JNK2 inhibitors and a new target of JNK2 to treat ALI and sepsis.

Design, synthesis, and bioactivity evaluation of novel amide/sulfonamide derivatives as potential anti-inflammatory agents against acute lung injury and ulcerative colitis.

The uneven regulation of inflammation is related to various diseases, making anti-inflammation a potential option for the development of novel therapies. In this study, we designed and synthesized a total of fifty-eight novel amide/sulfonamide derivatives based on our previously reported anti-inflammatory compounds. The anti-inflammatory activities of these compounds were evaluated upon LPS-stimulated J774A.1 cells. Compounds 11a, 11b, 11c, and 11d potently reduced the release of IL-6 and TNF-α, and decreased the mRNA level of cytokines in J774A.1 cells. The most active compound 11d with IC50 value of 0.61 µM for IL-6 inhibition, and 4.34 µM for TNF-α inhibition restored IκB α and inhibited the translocation of phosphorylated p65 into the nucleus. In vivo evaluation indicated that 11d improved LPS-induced ALI and alleviated DSS-induced ulcerative colitis in mice. In conclusion, these results suggested compound 11d can be a new lead structure for the development of anti-inflammatory drugs against ALI and ulcerative colitis.

[Temporal and Spatial Variations in Root-associated Fungi Associated with Pinus sylvestris var. mongolica in the Semi-arid and Dry Sub-humid Desertified Regions of Northern China].

To illuminate the ecological functions of root-associated fungi (RAF) and their interactions with host plants, we revealed the root-associated fungal diversity and community compositions of Pinus sylvestris var. mongolica involving natural forests and plantations (half-mature, nearly mature, and mature forests) in the Hulunbuir Desert, Horqin Desert, and Mu Us Desert and investigated the environmental driving factors (climatic condition and soil property). The results indicated that: 1 the diversity of RAF in the natural forests was significantly lower than that in plantations (P<0.05), and the values were highest in the Mu Us Desert. There was a distinct geographical distribution in the RAF community, but the influence of stand age was not significant (P>0.05). 2 The relative abundance of ectomycorrhizal fungi (50.49%) in natural forests was higher than that in plantations, such as Acephala, Mycena, and Suillus. The indicator genera were diverse involving the natural forests (Acephala) and plantations in the Hulunbuir Desert (Sarcodon), Horqin Desert (Russula and Calostoma), and Mu Us Desert (Geopora, Mallocybe, and Tomentella). 3 The indicator genera were mainly affected by available nitrogen content, available phosphorus content, and stand age, and few indicator genera were related to soil water content, pH, and total nitrogen content. A total of 43.25% of the variation in the RAF community was accounted for by both geographic location and environmental factors. Overall, geographic location and environmental factors shaped the spatial variation in the RAF structure and function of P. sylvestris natural forests and plantations in the semi-arid and dry sub-humid desertified regions; there were no significant temporal variations in RAF across stand ages, but the nonuniformity in fungal distribution with stand ageing cannot be ignored. The large population of symbiotic fungi in natural forests was conducive to the healthy growth of hosts; the ratio of symbiotic, saprophytic, and pathotrophic fungi varied in different plantations, and the increase in the proportion of saprophytic and pathotrophic fungi may have negative effects on the growth and health of plantations. This improved information will provide a theoretical basis for the management of P. sylvestris plantations.

Prognostic Role of Serum Soluble Tim-3 in Severe Traumatic Brain Injury: A Prospective Observational Study.

Objective: T cell immunoglobulin and mucin domain-3 (Tim-3) may be implicated in neuroinflammation. Herein, we attempted to discern the role of serum soluble (s) Tim-3 as an inflammatory prognostic biomarker of severe traumatic brain injury (sTBI). Methods: In this prospective observational study of 112 sTBI patients and 112 controls, serum sTim-3 levels were determined, Rotterdam computed tomography (CT) classification and Glasgow coma scale (GCS) were selected as the two severity indicators, serum C-reactive protein (CRP) was regarded as an inflammatory biomarker, and poor prognosis was referred to as extended Glasgow outcome scale (GOSE) scores 1-4 at 180 days after trauma. Results: Serum sTim-3 levels were markedly higher in patients than in controls (median, 4.2 ng/mL versus 0.7 ng/mL; P<0.001). Serum sTim-3 levels of patients were independently related to Rotterdam CT scores (ß=1.126), GCS scores (ß=-0.589), serum CRP levels (ß=0.155) and GOSE scores (ß=-0.211). Serum sTim-3 appeared as an independent predictor of post-traumatic 180-day mortality (odds ratio=1.289), overall survival (hazard ratio=1.208) and poor prognosis (odds ratio=1.293). Serum sTim-3 levels discriminated patients at risk of post-injury 180-day mortality and poor prognosis with areas under curve (AUCs) at 0.753 and 0.782, respectively. Serum sTim-3 levels combined with GCS scores and Rotterdam CT scores (AUC=0.869) exhibited significantly higher AUC than Rotterdam CT scores (P=0.026), but not than GCS scores (P=0.181) for death prediction and their combination (AUC=0.895) had significantly higher AUC than GCS scores (P=0.036) or Rotterdam CT scores (P=0.005) for outcome prediction. Conclusion: Elevated serum sTim-3 levels, in close correlation with traumatic severity and inflammation, are substantially associated with long-term death and poor outcome, indicating that serum sTim-3, as an inflammatory biomarker, may be of clinical significance in severity assessment and prediction of prognosis following sTBI.

Knockdown of Krüppel-Like Factor 9 Inhibits Aberrant Retinal Angiogenesis and Mitigates Proliferative Diabetic Retinopathy.

Advanced proliferative diabetic retinopathy (PDR) characterized by aberrant retinal angiogenesis is a leading cause of retinal detachment and blindness. Krüppel-like factor 9 (KLF9), a member of the zinc-finger family of transcription factors, participates in the development of diabetic nephropathy and the promotion of angiogenesis of human umbilical vein endothelial cells. Therefore, we speculate that KLF9 may exert a crucial role in PDR. The current study revealed that KLF9 was highly expressed in the high glucose (HG)-treated human retinal microvascular endothelial cells (HRMECs) and the retinas of oxygen-induced retinopathy (OIR) rats. Knockdown of KLF9 inhibited the proliferation, migratory capability, invasiveness and tube formation of HG-treated HRMECs. Besides, knockdown of KLF9 decreased the expression of yes-associated protein 1 (YAP1) in HG-treated HRMECs. Dual-luciferase reporter assays confirmed that KLF9 transcriptionally upregulated YAP1 expression. Overexpression of YAP1 reversed the KLF9 silencing-induced repression of HRMEC proliferation and tube formation. Further in vivo evidence demonstrated that knockdown of KLF9 reduced the expression of Ki67, CD31 and vascular endothelial growth factor A (VEGFA) in the retinas of OIR rats. Collectively, KLF9 silencing might mitigate the progression of PDR by inhibiting angiogenesis via blocking YAP1 transcription.

Acibenzolar-S-methyl activates calcium signalling to mediate lignin synthesis in the exocarp of Docteur Jules Guyot pears.

'Docteur Jules Guyot' pears were immersed in acibenzolar-S-methyl (ASM) and 0.01 mol L-1 ethyl glycol tetra acetic acid (EGTA) to investigate the changes of Ca2+ receptor proteins and phenylpropanoid pathway. Results showed that ASM treatment increased the activities of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate coenzyme A ligase (4CL), polyphenol oxidase (PPO), and cinnamyl alcohol dehydrogenase (CAD) in the exocarp of pears, whereas EGTA pre-treatment inhibited the activities of these enzymes. ASM treatment also enhanced the transcription of PcPAL, PcC4H, Pc4CL, PcC3H, PcCOMT, PcCCoAOMT, PcCCR, PcPOD, PcCDPK1, PcCDPK2, PcCDPK5, PcCDPK11, PcCDPK13, PcCBL1, PcCBL9, PcCIPK14, and PcCML27 in pears. EGTA + ASM treatments inhibited the transcription of PcPAL, PcC4H, Pc4CL, PcC3H, PcCCR, PcF5H, PcCAD, PcCDPK11, PcCDPK26, PcCDPK32, PcCBL1, PcCIPK14, PcCIPK23, and PcCaM in the fruit. All these results indicated that ASM induced the gene expressions of Ca2+ receptor proteins, the key enzyme activities and gene expressions in phenylpropanoid pathway; Ca2+ mediated phenylpropane metabolism in pears after ASM treatment.

Organic acids metabolism and GABA shunt involved in maintaining quality of Malus domestica by methyl jasmonate treatment.

Apples (cv. Golden Delicious) were used as the materials to investigate methyl jasmonate (MeJA) dipping on quality parameters, organic acids metabolism and GABA shunt during storage at 21 ± 1 °C and 75 ± 5 % relative humidity. Results demonstrated that MeJA treatment reduced mass loss, respiratory intensity and ethylene release, and maintained higher flesh firmness and soluble solid content of apples. MeJA also decreased malic acid content, increased succinic and tartaric acids contents, and inhibited cytoplasmic aconitase (Cyt-ACO), NADP-malate (NADP-ME), phosphoenolpyruvate dehydrogenase (PEPC), mitochondrial citrate synthase (Mit-CS), glutamate dehydrogenase (GAD), and GABA transferase (GABA-T) activities in apples. NADP-isocitrate dehydrogenase (NADP-IDH), mitochondrial cis-aconitase (Mit-ACO), and cytoplasmic NAD-malate dehydrogenase (CytNAD-MDH) activities in apples were also enhanced by MeJA dipping. Moreover, MeJA dipping enhanced MdCytNAD-MDH and MdNADP-IDH expressions, and down-regulated MdGAD, MdGABA-T, MdNADP-ME, MdPEPC, MdCyt-ACO and MdMit-CS expressions in apples. These results suggest that MeJA dipping can maintain storage quality of "Golden Delicious" apples by regulating organic acids metabolism and GABA shunt.

Trehalose Regulates Starch, Sorbitol, and Energy Metabolism to Enhance Tolerance to Blue Mold of "Golden Delicious" Apple Fruit.

The efficacy of trehalose on the lesion diameter of apples (cv. Golden Delicious) inoculated with Penicillium expansum was evaluated to screen the optimal concentration. The changes in gene expression and activity of the enzyme in starch, sorbitol, and energy metabolism were also investigated in apples after trehalose treatment. The results revealed that trehalose dipping reduced the lesion diameter of apples inoculated with P. expansum. Trehalose suppressed the activities and gene expressions of ß-amylase, NAD-sorbitol dehydrogenase, and NADP-sorbitol dehydrogenase, whereas it decreased the sorbitol 6-phosphate dehydrogenase gene expression and amylose, amylopectin, total starch, and reducing sugar contents. Additionally, trehalose improved the gene expressions and activities of α-amylase, starch-branching enzymes, total amylase, H+-ATPase, and Ca2+-ATPase, as well as soluble sugar, adenosine triphosphate, and adenosine diphosphate contents and energy charge in apples. These findings imply that trehalose could induce tolerance to the blue mold of apple fruit by regulating starch, sorbitol, and energy metabolism.

Retinal Src homology region 2-containing protein tyrosine phosphatase 2 silencing alleviates diabetic retinopathy via suppressing inflammatory response and oxidative stress by regulating Yes-associated protein 1 activity.

Diabetic retinopathy (DR) is the prevalent microvascular complication of diabetes mellitus (DM), and it may lead to permanent blindness. The previous publication has indicated that both inflammatory response and oxidative stress are critical factors involved in DR progression, however, the accurate regulatory mechanism remains to be revealed. Src homology region 2 (SH2)-containing protein tyrosine phosphatase 2 (SHP2), a member of the protein tyrosine phosphatase family, was reported to play a role in diabetic nephropathy, whereas its function in DR was unknown and required further exploration. The level of phosphorylated, not the total, SHP2 increased in the retinas of rats with streptozotocin injection-induced DM. Further, the intravitreal injection of SHP2 shRNA lentivirus alleviated retinal pathological changes, and inhibited inflammatory response and oxidative stress, which were accompanied with Yes-associated protein 1 (YAP1) deactivation in DR rats. Additional co-immunoprecipitation results confirmed the interaction of SHP2 and YAP1. Collectively, our data preliminarily show that DR amelioration-induced by SHP2 inhibition in rats may attribute to the deactivation of YAP1 pathway.

Shielded geomagnetic field accelerates glucose consumption in human neuroblastoma cells by promoting anaerobic glycolysis.

A shielded geomagnetic field, also called the hypomagnetic field (HMF), interferes with the metabolic processes of various cells and animals exhibiting diverse effects in different models, however, its underlying mechanism remains largely unknown. In this study, we assessed the effect on the energy metabolism of SH-SY5Y cells in HMF and found that HMF-induced cell proliferation depends on glucose supply. HMF promoted SH-SY5Y cell proliferation by increasing glucose consumption rate via up-regulating anaerobic glycolysis in the cells. Increased activity of LDH, a key member of glycolysis, was possibly a direct response to HMF-induced cell proliferation. Thus, we unveiled a novel subcellular mechanism underlying the HMF-induced cellular response: the up-regulation of anaerobic glycolysis and repression of oxidative stress shifted cellular metabolism more towards the Warburg effect commonly observed in cancer metabolism. We suggest that cellular metabolic profiles of various cell types may determine HMF-induced cellular effects, and a magnetic field can be applied as a non-invasive regulator of cell metabolism.

Postharvest melatonin dipping maintains quality of apples by mediating sucrose metabolism.

Senescence is a pivotal factor that causes quality breakdown and economic loss of fruit after harvest. In this study, 'Golden Delicious' apples were used as the materials to investigate the effect of melatonin dipping on quality parameters and sucrose metabolism during room temperature storage. Postharvest melatonin treatment inhibited respiratory intensity and ethylene release, increased flesh firmness, soluble sugar, ascorbic acid, and soluble solid contents, and titratable acid in apples. Furthermore, melatonin treatment inhibited acid invertase and neutral invertase activities, increased sucrose synthase and sucrose phosphate synthase activities, and repressed the activities of sorbitol dehydrogenase, sorbitol oxidase and sucrose synthase cleavage in apple fruit. All these findings suggest that exogenous application of melatonin could maintain quality of 'Golden Delicious' apples by mediating the enzyme activity in sucrose metabolism.

[Effect of electroacupuncture of "Zhiyang" (GV9) and "Jizhong" (GV6) on expression of CGRP and NLRP3 in rats with spinal cord injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zhiyang" (GV9) and "Jizhong" (GV6) of the Governor Vessel on the expression of calcitonin gene related peptide (CGRP)and NOD-like receptor pyrin containing 3 (NLRP3) in the injured anterior horn (AH) of rats with spinal cord injury (SCI), so as to explore its mechanisms underlying improvement of SCI. METHODS: Thirty-six male SD rats were randomly divided into sham operation (sham) group, model group and EA group which were further randomly allocated to 7 day (d) and 14 d subgroups (n=6 per subgroup). The SCI model was established by clamping the exposed spinal cord with an aneurysm clip. Rats of the EA group received EA of GV6 and GV9 for 30 min, once daily for 7 and 14 days, respectively. The Basso, Beattie and Bresnahan (BBB) was used to assess changes of the locomotor function on the 1st, 3rd, 7th and 14th day after SCI. The protein expression levels of CGRP, NLRP3, apoptosis-associated speck-like protein (ASC) and cysteinyl aspartate specific protease-1 (caspase-1) in the anterior horn region of the spinal cord on the 7th and 14th after SCI were detected by Western blot. RESULTS: The BBB scores on the 1st, 3rd, 7th and 14th d after SCI were significantly lower in the model group than in the sham group (P<0.05), whereas the expression levels of NLRP3, ASC and caspase-1 proteins on day 7 and 14 were markedly higher in the model group than in the sham group (P<0.05, P<0.01). Compared with the model group, the BBB score and CGRP expression on the 7th and 14th d were significantly increased in the EA group (P<0.05,P<0.01), while the expression levels of NLRP3, ASC and caspase-1 were obviously decreased in the EA group (P<0.01). CONCLUSION: EA of GV6 and GV9 can improve the locomotion of SCI rats, which may be associated with its function in up-regulating the expression of CGRP and down-regulating the expression of NLRP3, ASC and caspase-1 proteins in the spinal anterior horn tissue.

Serum hypoxia-inducible factor 1alpha emerges as a prognostic factor for severe traumatic brain injury.

BACKGROUND: Hypoxia-inducible factor 1alpha (HIF-1α) is implicated in the cell's response to hypoxia. We investigated whether serum HIF-1α concentrations are correlated with the severity and clinical outcome of severe traumatic brain injury (sTBI). METHODS: Serum HIF-1α concentrations were quantified in 104 sTBI patients and 80 healthy controls. Trauma severity was assessed using Glasgow coma scale (GCS). Glasgow outcome scale (GOS) score of 1-3 at post-trauma 90 days was defined as a poor outcome. Multivariate analyses were performed to discern the relationship between serum HIF-1α concentrations and outcome. RESULTS: Patients displayed significantly higher serum HIF-1α concentrations than controls (median, 294.9 versus 102.7 pg/ml). HIF-1α concentrations were intimately related to GCS scores (r = -0.62) and GOS scores (r = -0.64). 48 patients (46.2%) experienced a poor outcome. Serum HIF-1α concentrations > 280.2 pg/ml significantly distinguished patients with the development of poor outcome with 77.1% sensitivity and 69.6% specificity (AUC, 0.750; 95% CI: 0.655-0.829). Serum HIF-1α concentrations > 280.2 pg/ml emerged as an independent predictor for poor outcome (OR: 4.179; 95% CI: 1.024-17.052). CONCLUSIONS: Serum HIF-1α concentrations are tightly associated with trauma severity and poor 90-day outcome, substantializing serum HIF-1α as a promising prognostic biomarker for sTBI.

Glucomannan from Aloe vera Gel Promotes Intestinal Stem Cell-Mediated Epithelial Regeneration via the Wnt/ß-Catenin Pathway.

Intestinal stem cells (ISCs) are essential to maintain intestinal epithelial regeneration and barrier function. Our previous work showed that glucomannan from Aloe vera gel (AGP) alleviated epithelial damage, but the mechanism was still elusive. Herein, RNA-sequencing analysis showed that proliferation and differentiation of intestinal epithelial cells as well as the canonical Wnt pathway were involved in this process. Further experiments exhibited that AGP promoted nuclear translocation of ß-catenin and expression of transcription factor 7, increased the number of Lgr5+ ISCs, and differentiated epithelial cells in mice colon. Intriguingly, AGP reversed the inhibition of IEC-6 cells proliferation induced by an inhibitor of the canonical Wnt pathway. Hence, this study implied that AGP promoted proliferation and differentiation of colon stem cells via Wnt/ß-catenin signaling, which subsequently facilitated the regeneration of epithelial cells and alleviated colitis in mice. It may provide new insights into the role of polysaccharides in regulating intestinal homeostasis and relieving intestinal injury.

The effect of various urban design parameter in alleviating urban heat island and improving thermal health-a case study in a built pedestrianized block of China.

Increasing urban heat island and global warming have aroused serious thermal environmental problems and even harm people's thermal health. Because of the importance in people's daily life, a commercial pedestrianized block represents a symbol of a city or metropolis; therefore, focusing the attention on the thermal environment in such regions is very necessary. Most of the researches on the urban thermal environment are calculated by remote sensing data; limited by the low spatial resolution of remote sensing image, it may not obviously reflect the true thermal environment of the research site, especially in some microscale regions. Based on this, the new software ENVI-met is developed to research the thermal environment and forecast people's thermal sensation in a microscale region. Therefore, the objective of this study aims at conducting field measurement and numerical simulation to assess the thermal environment of a typical commercial pedestrianized space in southern China and assess the different urban design parameters in ameliorating the urban heat island effect. Our final results demonstrate a quantitative evidence for establishing a comprehensive standard for improving the thermal environment in a microscale region, and this study also can be a supplementary in the research field about improving thermal health.

Lysosome-Mediated Cytotoxic Autophagy Contributes to Tea Polysaccharide-Induced Colon Cancer Cell Death via mTOR-TFEB Signaling.

Targeting autophagy and lysosome may serve as a promising strategy for cancer therapy. Tea polysaccharide (TP) has shown promising antitumor effects. However, its mechanism remains elusive. Here, TP was found to have a significant inhibitory effect on the proliferation of colon cancer line HCT116 cells. RNA-seq analysis showed that TP upregulated autophagy and lysosome signal pathways, which was further confirmed through experiments. Immunofluorescence experiments indicated that TP activated transcription factor EB (TFEB), a key nuclear transcription factor modulating autophagy and lysosome biogenesis. In addition, TP inhibited the activity of mTOR, while it increased the expression of Lamp1. Furthermore, TP ameliorated the lysosomal damage and autophagy flux barrier caused by Baf A1 (lysosome inhibitor). Hence, our data suggested that TP repressed the proliferation of HCT116 cells by targeting lysosome to induce cytotoxic autophagy, which might be achieved through mTOR-TFEB signaling. In summary, TP may be used as a potential drug to overcome colon cancer.

Efficiency analysis by mfERG and OCT of intravitreal injection with ranibizumab on diabetic macular edema.

AIM: To analyze the clinical efficacy of intravitreal injection of ranibizumab (IVR) on diabetic macular edema (DME) with multifocal electroretinography (mfERG) and optical coherence tomography (OCT). METHODS: A total of 41 patients (41 eyes) with DME were treated with IVR. The best corrected visual acuity (BCVA), mfERG results, and OCT were analyzed to compare to the baselines at 1wk, 1, and 3mo after operation. RESULTS: The BCVA was significantly improved in all eyes at each time point (P<0.001). The macular area leakage and edema were reduced 1wk and 1mo after IVR, and the central fovea thickness (CFT) was significantly reduced compared to baseline (P<0.001). The mfERG, two-dimensional and three-dimensional images all showed that the macular fovea (1 ring) response density decreased, and the fovea and macular area spikes significantly decreased or disappeared. The amplitude density of the P1 wave was increased, and the latency of the P1 wave was shortened than preoperation (P<0.001). At 1wk and 1mo after the operation, there was a negative correlation between the amplitude density of P1 waves and CFT. CONCLUSION: OCT and mfERG fully demonstrate the importance of IVR for DME patients from the macular morphology and function, especially the significance of mfERG in this disease.