Proteomics identifies hypothermia induced adiponectin protects corneal endothelial cells via AMPK mediated autophagy in phacoemulsification.

AIM: To explore the molecular mechanism underlying the protective effect of hypothermic perfusion on the corneal endothelium during phacoemulsification. METHODS: Phacoemulsification was performed on New Zealand white rabbits. Perfusate at different temperatures was used during the operation, and the aqueous humor was collected for proteomic sequencing after the operation. Corneal endothelial cell injury was simulated by a corneal endothelial cell oxygen-glucose deprivation/reoxygenation (OGD/R) model in vitro. Flow cytometry and evaluation of fluorescent LC3B puncta were used to detect apoptosis and autophagy, and western blotting was used to detect protein expression. RESULTS: A total of 381 differentially expressed proteins were identified between the two groups. In vitro, 4 ℃ hypothermia significantly reduced apoptosis and promoted autophagy. Apoptosis increased after autophagy was inhibited by 3-Methyladenine (3-MA). Furthermore, adiponectin (ADIPOQ) knockdown inhibited phospho-AMPK and blocked the protective effect of hypothermia on corneal endothelial cells. CONCLUSIONS: We investigated the differential expression of proteins between the hypothermia group and normothermia group by proteomics. Moreover, hypothermia-induced ADIPOQ can reduce apoptosis by promoting AMPK-mediated autophagy.

Comprehensive evaluation of functional vision, quality of life, and cognitive ability in pediatric uveitis.

BACKGROUND: Pediatric uveitis may cause severe impairment of vision in children and affect their quality of life as well as cognitive ability. This study aims to evaluate the functional vision, visual-related and health-related quality of life, and cognitive ability in pediatric uveitis. METHODS: Children with uveitis aged 5-16 years old completed six validated instruments to assess functional visual ability with Cardiff Visual Ability Questionnaire for Children (CVAQC), vision-related quality of life with Impact of Vision Impairment for Children (IVI-C), health-related quality of life with Pediatric Quality of Life Inventory (PedsQL), cognitive ability with Chinese Wechsler Intelligence Scale for Children (C-WISC), and depression and anxiety evaluation with Hospital Anxiety and Depression Scale (HAD). RESULTS: The CVAQC, IVI-C, and PedsQL scores of pediatric uveitis were significantly lower than that of normal levels. Full-scale intelligence quotient (IQ) and performance IQ were significantly lower in pediatric uveitis patients with impaired vision in their best eye (visual acuity < 0.3) compared to those with a vision equal to or better than 0.3. Verbal IQ was significantly lower in male pediatric uveitis patients with impaired vision compared to those with a vision equal to or better than 0.3. Additionally, parents of pediatric uveitis patients with impaired vision generally had lower educational levels than parents of those with a vision equal to or better than 0.3. CONCLUSIONS: Impaired vision caused by pediatric uveitis has a significant impact on children's functional visual ability and quality of life. The development of cognitive function in pediatric uveitis is also significantly hindered.

[Research progress of Shegan Mahuang Decoction and predictive analysis on its Q-markers].

Shegan Mahuang Decoction has been used in clinical practice for thousands of years, and is a classical formula for treating asthma and other respiratory diseases, with the effects of ventilating lung, dispersing cold, and relieving cough and asthma. This paper summarized the history, clinical application and mechanism of Shegan Mahuang Decoction, and predicted its quality markers(Q-markers) based on the "five principles" of Q-markers. The results suggested that irisflorentin, tectoridin, tectorigenin, irigenin, ephedrine, pseudoephedrine, asarinin, methyleugenol, shionone, epifriedelanol, tussilagone, 6-gingerol, trigonelline, cavidine, schizandrin, and schizandrin B could be used as Q-markers of Shegan Mahuang Decoction, which provided a basis for the quality control and subsequent research and development of Shegan Mahuang Decoction.

[Improvement effect of Shegan Mahuang Decoction on rats with cold-induced asthma based on TRPV1/NRF-1/mtTFA pathway].

This study investigated the therapeutic effect of Shegan Mahuang Decoction(SGMHD) on cold-induced asthma in rats and explored its underlying mechanism. Seventy-two healthy male SD rats of specific pathogen free(SPF) grade were randomly divided into a blank group, a model group, a positive control group(dexamethasone, 0.4 mg·kg~(-1)), and low-, medium-, and high-dose SGMHD groups(3.2, 6.4, and 12.8 g·kg~(-1)). The blank group received saline, while the other groups were sensitized by intraperitoneal injection of ovalbumin(OVA) solution. Subsequently, the rats were placed in a cold chamber adjustable to 0-2 ℃, and OVA solution was ultrasonically nebulized to induce cold-induced asthma in rats. After three weeks of treatment, the general behaviors of rats were observed. Hematoxylin-eosin(HE) staining was used to evaluate pathological changes in lung tissues, periodic acid-Schiff(PAS) staining assessed mucin changes, and Masson staining was performed to examine collagen deposition. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of the inflammatory factors interleukin-4(IL-4) and vascular endothelial growth factor(VEGF) in serum and bronchoalveolar lavage fluid(BALF). Real-time quantitative polymerase chain reaction(RT-PCR) was employed to assess the mRNA expression levels of transient receptor potential vanilloid subfamily member 1(TRPV1), nuclear respiratory factor 1(NRF-1), and mitochondrial transcription factor A(mtTFA) in lung tissues. Western blot was used to measure the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues. Compared with the blank group, the model group exhibited signs of rapid respiration, increased frequency of defecation with looser stools, and disheveled and dull fur. Pathological results showed significant infiltration of inflammatory cells in lung tissues, narrowing of bronchial lumens, increased mucin secretion, and enhanced collagen deposition in the model group. Additionally, the levels of IL-4 and VEGF in serum and BALF were significantly elevated, and the mRNA and protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were significantly increased. Compared with the model group, SGMHD improved the behaviors of rats, alleviated pathological changes in lung tissues, mucin production, and collagen deposition, significantly decreased the levels of IL-4 and VEGF in serum and BALF, and reduced the mRNA expression levels of TRPV1, NRF-1, and mtTFA in lung tissues, with the medium-dose SGMHD group showing the most significant effect. Moreover, the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were also reduced, with the medium-dose SGMHD group exhibiting the most significant effect. In conclusion, this study demonstrates that SGMHD can alleviate airway inflammation and inhibit airway remodeling in cold-induced asthma rats. These effects may be associated with the modulation of the TRPV1/NRF-1/mtTFA signaling pathway.

[Research progress of Shenling Baizhu San and predictive analysis on its quality markers].

Shenling Baizhu San is a classic prescription for replenishing Qi to invigorate the spleen and dispelling dampness to check diarrhea, which mainly treats the syndrome of spleen deficiency and heavy dampness. With the pharmacological effects of regulating immune system, improving lung function and gastrointestinal function, and resisting oxygen, tumor, and inflammation, Shenling Baizhu San is commonly used in modern clinical practice to treat chronic obstructive pulmonary disease, pulmonary fibrosis, bronchial asthma, irritable bowel syndrome, ulcerative colitis, chronic diarrhea, and diabetic, etc. This paper summarized the chemical constituents, pharmacological effects, and clinical application of Shenling Baizhu San in recent years, and predictively analyzed the quality markers of Shenling Baizhu San according to the "five principles" of Q-marker. The Q-markers of Shenling Baizhu San involved ginsenoside Rg_1, ginsenoside Re, ginsenoside Rb_1, pachymic acid, dehydrotumulosic acid, batatasin Ⅰ, batatasin Ⅲ, diosgenin, liensinine, neferine, luteolin, quercetin, glycerol trioleate, ß-sitosterol, platycodin D, glycyrrhizic acid, glycyrrhetinic acid, liquiritin, pipecolinic acid, atractylenolide Ⅰ, atractylenolide Ⅲ, and bornyl acetate, which provided references for the quality control and follow-up research of Shenling Baizhu San.

Inhibition of histamine receptor H3 suppresses the growth and metastasis of human non-small cell lung cancer cells via inhibiting PI3K/Akt/mTOR and MEK/ERK signaling pathways and blocking EMT.

Recent evidence shows that the expression levels of histamine receptor H3 (Hrh3) are upregulated in several types of cancer. However, the role of Hrh3 in non-small cell lung cancer (NSCLC) has not been elucidated. In the present study, we showed that the expression levels of Hrh3 were significantly increased in NSCLC samples, and high levels of Hrh3 were associated with poor overall survival (OS) in NSCLC patients. In five human NSCLC cell lines tested, Hrh3 was significantly upregulated. In NSCLC cell lines H1975, H460, and A549, Hrh3 antagonist ciproxifan (CPX, 10-80 µM) exerted moderate and concentration-dependent inhibition on the cell growth and induced apoptosis, whereas its agonist RAMH (80 µM) reversed these effects. Furthermore, inhibition of Hrh3 by CPX or siRNA retarded the migration and invasion of NSCLC cells through inhibiting epithelial-mesenchymal transition (EMT) progression via reducing the phosphorylation of PI3K/Akt/mTOR and MEK/ERK signaling pathways. In nude mice bearing H1975 cell xenograft or A549 cell xenograft, administration of CPX (3 mg/kg every other day, intraperitoneal) significantly inhibited the tumor growth with increased E-cadherin and ZO-1 expression and decreased Fibronectin expression in tumor tissue. In conclusion, this study reveals that Hrh3 plays an important role in the growth and metastasis of NSCLC; it might be a potential therapeutic target against the lung cancer.

Crossed versus conventional pseudophakic monovision for high myopic eyes: a prospective, randomized pilot study.

BACKGROUND: Aiming at spectacle independence, conventional pseudophakic monovision has been widely used in myopia patients with bilateral monofocal intraocular lens implantation. However, the crossed monovision, which is to correct the dominant eye for near vision and the non-dominant eye for distant vision, has been mentioned preferable for high myopic cataract patients by some studies. We have conducted this study to compare clinical results to assess the feasibility of conventional and crossed monovision for high myopic pseudophakic patients by comparing patient satisfaction, visual function and spectacle independence. METHOD: Forty-sixth high myopia patients were divided into two groups: 22 in crossed monovision group with patients whose refraction targeted to - 2.00 diopters (D) in the dominant eye and - 0.50D in the non-dominant eye; 24 in conventional monovision group with patients whose refraction targeted to - 0.50D in the dominant eye and - 2.00D in the non-dominant eye. Binocular uncorrected distance visual acuity (BUDVA), binocular uncorrected near visual acuity (BUNVA), binocular corrected distant visual acuity (BCDVA), binocular corrected near visual acuity (BCNVA), contrast visual acuity and stereoacuity were examined at postoperative 2 weeks, 1 month and 3 months. Questionnaires were completed by patients 3 months after binocular surgery to evaluate patients' satisfaction and spectacle independence. RESULTS: The conventional monovision and the crossed monovision group showed no significant differences of mean BUDVA, BUNVA, BCDVA, BCNVA 2 weeks, 1 month or 3 months postoperatively (P > 0.05). There was no difference in the bilateral contrast sensitivity or stereoscopic function between the convention conventional and crossed monovision groups (P > 0.05). Patient satisfaction with near and distant vision, as well as spectacle dependence did not differ significantly between the two groups (P > 0.05). CONCLUSION: Crossed pseudophakic monovision exhibited similar visual function when compared with conventional monovision technique, which indicates that it is an effective option to improve the visual functionality and quality of life for high myopic patients who considering bilateral cataract surgery. TRIAL REGISTRATION: The Institutional Review Board and Ethics committee of the First Affiliated Hospital of Chongqing Medical University, Chongqing, China. The trial registration was submitted in September 2018 and passed on March 18, 2020, and the registration number is: ChiCTR2000030935 .

Effect of hypothermic perfusion on phacoemulsification in cataract patients complicated with uveitis: a randomised trial.

BACKGROUND: To evaluate the effectiveness and safety of hypothermic perfusion in the phacoemulsification of cataract caused by uveitis. METHODS: This was a prospective, single-masked, randomised, controlled clinical trial. One hundred and six patients with uveitis-associated cataract underwent phacoemulsification with perfusion fluid temperature at 4 °C (treatment group) or 24 °C (control group). Anterior chamber inflammation grade, corneal endothelial cell count, corneal thickness, macular fovea thickness, and intraocular pressure (IOP) were observed on the 1st day and 7th day after operation. RESULTS: The aqueous flare score was 0.83 ± 0.76 in the 4 °C group, which was lower than that in the 24 °C group (1.51 ± 1.02, p = 0.006) on the first day after operation. The aqueous cells score was lower in the 4 °C group (0.17 ± 0.38) than that in the 24 °C group (0.62 ± 0.94, p = 0.025). The mean corneal thickness of incision in the 4 °C group (907.66 ± 85.37 µm) was thinner than that in the 24 °C group (963.75 ± 103.81 µm, p = 0.005). Corneal endothelial cells density, macular fovea thickness, or percentage of transiently increased IOP showed no difference between the two groups (p > 0.05). There was no significant difference in all the main outcome parameters between the two groups on the 7th day after operation (p > 0.05). CONCLUSIONS: Hypothermic perfusion in the phacoemulsification of uveitis-associated cataract is safe, and it can effectively inhibit anterior chamber inflammation and reduce the incisional corneal edema in the early postoperative stage. TRIAL REGISTRATION: The study was registered with the Chinese Clinical Trial Registry. (http://www.chictr.org.cn/, Registration Number: ChiCTR1800016145).

RNAi of the nuclear receptor HR3 suggests a role in the molting process of the spider mite Panonychus citri.

Ecdysteroids regulate molting in arthropods by binding to heterodimers of the ecdysone receptor and retinoid-X-receptor, homologous to the ultraspiracle protein, to induce the expression of downstream signal response genes including the nuclear receptor HR3. However, the detailed expression dynamics of HR3 during molting in spider mites are not yet clear. In this study, the full length of PcHR3 was retrieved based on the genome of citrus red mite, Panonychus citri. The open reading frame is 1707 bp encoding 568 amino acids, which contains a DNA binding domain and a ligand binding domain. Then, the expression pattern of PcHR3 was analyzed throughout the development of the deutonymph by RT-qPCR. The result showed that PcHR3 was mainly transcribed in the late deutonymph stage, when the deutonymph was at least 24 h old and motionless, the critical point at which the mites started molting. Transcription reached the highest level in 32-h-old deutonymphs and decreased by 36 h, where the mites remained in a quiescent state. Further silencing of PcHR3 by leaf-disc-based delivery of dsRNA to 8-h-old deutonymph mites, resulted in retarded development and death of 58% of deutonymphs. In summary, we suggest that PcHR3 regulates the latter stages of molting in P. citri.

Expression dynamics of key ecdysteroid and juvenile hormone biosynthesis genes imply a coordinated regulation pattern in the molting process of a spider mite, Tetranychus urticae.

In insects, the ecdysteroid 20-hydroxyecdysone coordinates with juvenile hormone (JH) to regulate the process of molting, development and metamorphosis; however, this interaction is still unclear in the mites. In this study, we investigated the gene related to ecdysteroid and JH biosynthesis pathways, including four ecdysteroid and 11 JH biosynthesis genes. We examined their expression patterns during molting of different developmental stages of the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), an important agricultural pest that feeds on more than 1100 plant species. The expression of ecdysteroid biosynthesis Halloween genes exhibited a positive zigzag-like pattern, with a peak after 8 h of molting and a drop 8 h after entering each quiescent stage. In contrast, JH biosynthesis genes expression displayed a negative zigzag-like pattern, with a peak at 8 h after entering each quiescent stage and a drop after 8 h of each molting. These opposite patterns imply that ecdysteroid and JH expression is coordinated during the developmental transition. Our data provide an initial perspective on the co-expression of ecdysteroid and JH biosynthesis genes to regulate this important developmental process in the two-spotted spider mite.

miRNA-133b targets FGFR1 and presents multiple tumor suppressor activities in osteosarcoma.

BACKGROUND: Osteosarcoma (OS) is the most common bone malignancy prevalent in children and young adults. MicroRNA-133b (miR-133b), through directly targeting the fibroblast growth factor receptor 1 (FGFR1), is increasingly recognized as a tumor suppressor in different types of cancers. However, little is known on the biological and functional significance of miR-133b/FGFR1 regulation in osteosarcoma. METHODS: The expressions of miR-133b and FGFR1 were examined by RT-qPCR and compared between 30 paired normal bone tissues and OS tissues, and also between normal osteoblasts and three OS cells lines, MG-63, U2OS, and SAOS-2. Using U2OS and MG-63 as the model system, the functional significance of miR-133b and FGFR1 was assessed on cell viability, proliferation, apoptosis, migration/invasion, and epithelial-mesenchymal transition (EMT) by overexpressing miR-133b and down-regulating FGFR1 expression, respectively. Furthermore, the signaling cascades controlled by miR-133b/FGFR1 were examined. RESULTS: miR-133b was significantly down-regulated while FGFR1 robustly up-regulated in OS tissues and OS cell lines, when compared to normal bone tissues and normal osteoblasts, respectively. Low miR-133b expression and high FGFR1 expression were associated with location of the malignant lesion, advanced clinical stage, and distant metastasis. FGFR1 was a direct target of miR-133b. Overexpressing miRNA-133b or knocking down FGFR1 significantly reduced the viability, proliferation, migration/invasion, and EMT, but promoted apoptosis of both MG-63 and U2OS cells. Both the Ras/MAPK and PI3K/Akt intracellular signaling cascades were inhibited in response to overexpressing miRNA-133b or knocking down FGFR1 in OS cells. CONCLUSION: miR-133b, by targeting FGFR1, presents a plethora of tumor suppressor activities in OS cells. Boosting miR-133b expression or reducing FGFR1 expression may benefit OS therapy.

Comprehensive Evaluation of Soil Near Uranium Tailings, Beishan City, China.

To evaluate the impact of uranium tailings on soil composition and soil microbial, six soil samples at different distance from the uranium tailings (Beishan City, China) were collected for further analysis. Concentrations of radionuclides (238U and 232Th), heavy metals (Mn, Cd, Cr, Ni, Zn, and Pb) and organochlorine pesticide were determined by ICP-MS and GC, they were significantly higher than those of the control. And the Average Well Color Development as well as the Shannon, the Evenness, and the Simpson index were calculated to evaluate the soil microbial diversity. The carbon utilization model of soil microbial community was also analyzed by Biolog-eco. All results indicated that uranium tailings leaded to excessive radionuclides and heavy metals, and decreased the diversity of the soil microbial community. Our study will provide a valuable basis for soil quality evaluation around uranium tailing repositories and lay a foundation for the management and recovery of uranium tailings.

Heavy Metals in Sediment from Bei Shan River: Distribution, Relationship with Soil Characteristics and Multivariate Assessment of Contamination Sources.

The concentrations and correlation between some heavy metals (Mn, Cd, Cr, Cu, Ni, Zn and Pb) measured in sediments in part of Bei shan River near uranium mill tailings were studied. The mean concentration of Cr (57.7 mg/g) was more than the mean values established for uncultivated areas worldwide (46.3 mg/g). Negative correlations with pH and positive correlations with organic matter have been observed for most of elements analyzed in this study. Correlation analysis showed that all metals except Cr are highly correlated (p ≤ 0.01, p ≤ 0.05). This may indicate the different origin or controlling factors of Cr in analyzed sediments. Cluster analysis highlighted the lithogenic origin of heavy metals (Mn, Cd, Cu, Ni, Zn and Pb) and pointed out the primary input of Cr from anthropogenic sources.

Cross-resistance, inheritance and biochemical mechanism of abamectin resistance in a field-derived strain of the citrus red mite, Panonychus citri (Acari: Tetranychidae).

BACKGROUND: The citrus red mite, Panonychus citri (McGregor), a global pest of citrus, has developed different levels of resistance to various acaricides in the field. Abamectin is one of the most important insecticides/acaricides worldwide, targetting a wide number of insect and mite pests. The evolution of abamectin resistance in P. citri is threatening the sustainable use of abamectin for mite control. RESULTS: The abamectin resistant strain (NN-Aba), derived from a field strain NN by consistent selection with abamectin, showed 4279-fold resistance to abamectin compared to a relatively susceptible strain (SS) of P. citri. Cross-resistance of NN-Aba was observed between abamectin and emamectin benzoate, pyridaben, fenpropathrin and cyflumetofen. Inheritance analyses indicated that abamectin resistance in the NN-Aba strain was autosomal, incompletely recessive and polygenic. The synergy experiment showed that abamectin toxicity was synergized by piperonyl butoxide (PBO), diethyl maleate (DEM) and tributyl phosphorotrithiotate (TPP) in the NN-Aba strain, and synergy ratios were 2.72-, 2.48- and 2.13-fold, respectively. The glutathione-S-transferases activity in the NN-Aba strain were significantly increased by 2.08-fold compared with the SS strain. CONCLUSION: The abamectin resistance was autosomal, incompletely recessive and polygenic in P. citri. The NN-Aba strain showed cross-resistance to various acaricides with different modes of action. Metabolic detoxification mechanism participated in abamectin resistance in NN-Aba strain. These findings provide useful information for resistance management of P. citri in the field. © 2023 Society of Chemical Industry.

Sublethal and transgenerational effects of broflanilide on the citrus red mite, Panonychus citri.

BACKGROUND: The citrus red mite, Panonychus citri is a serious pest of the citrus industry and has developed resistance to many acaricides. Broflanilide is a novel meta-diamide insecticide that binds to a new site on the γ -aminobutyric acid receptor with high potency against pests. However, little information has been reported about its effect on the citrus red mite. RESULTS: Broflanilide exhibited higher toxicity to female adults and eggs of a laboratory strain of P. citri The median lethal concentration (LC50), 9.769 mg/L and 4.576 mg/L, respectively) than other commonly used acaricides and was also toxic to two P. citri field strains. Broflanilide treatment with LC10, LC20, and LC30 significantly decreased the fecundity and longevity of female adults of F0 P. citri compared with the control. The duration of larva, protonymph, deutonymph and adult, and total life span in the F1 generation were significantly reduced after treatment of F0 with broflanilide. Population parameters, including the intrinsic rate of increase (r) and finite rate of increase (λ), were significantly increased, and the mean generation time (T) of F1 progeny was significantly reduced in the LC20 treatment. The predicted population size of F1 increased when parental female adults were treated with sublethal concentrations. CONCLUSION: Broflanilide had high acaricidal activity toward P. citri, and exposure to a sublethal concentration significantly inhibited the population growth of F0. The transgenerational hormesis effect is likely to cause population expansion of F1. More attention should be paid when broflanilide is applied to control P. citri in citrus orchards. © 2024 Society of Chemical Industry.

Resistance of Panonychus citri (McGregor) (Acari: Tetranychidae) to pyridaben in China: monitoring and fitness costs.

BACKGROUND: Panonychus citri is a major citrus pest worldwide. The short life cycle and high reproductive potential of P. citri, combined with heavy acaricide use, have led to high levels of resistance to acaricides, posing a threat to global resistance management programs. Here, resistance monitoring was established to determine the pyridaben resistance status of ten P. citri populations in China from 2014 to 2021 using a leaf-dipping assay. Four characterized strains-the susceptible strain (Lab_S), the resistant strain (Pyr_R), as well as the segregated resistant strain (Pyr_Rs) and the segregated susceptible strain (Pyr_Control) derived from the crossing of the Lab_S and Pyr_R strains, were used to evaluate the life-history characteristics using age-stage, two-sex life tables. RESULTS: Most P. citri populations developed high resistance to pyridaben. Resistance levels exceeded 1000-fold in Yuxi, Anyue, Nanning, and Ganzhou populations compared with the Lab_S strain. Compared with Pyr_Control, two key fitness cost criteria, developmental period and fecundity, showed significant differences in Pyr_Rs under consistent conditions. The intrinsic rate of increase, net reproductive rate and gross reproductive rate were lower in the resistant strain compared with the Pyr_Control strain. The Pyr_Rs strain had a lower relative fitness of 0.934 compared with the Pyr_Control. Moreover, the life-history traits and population parameters of the Pyr_R strain also showed significant differences compared with the Lab_S strain. CONCLUSION: The resistance levels to pyridaben varied greatly among the different P. citri populations and showed regional differences. Substantial fitness costs are associated with pyridaben resistance. This study provides potential implications for developing strategies for resistance management in P. citri. © 2022 Society of Chemical Industry.

Molting process revealed by the detailed expression profiles of RXR1/RXR2 and mining the associated genes in a spider mite, Panonychus citri.

Spider mites have one ecdysone receptor (EcR) and multiple retinoid X receptors (RXRs). However, the function of these RXRs in spider mite development is unknown. Here, we screened the expression dynamics of two PcRXR isoforms at 4 h intervals in the deutonymphal stage of Panonychus citri. The results showed that PcEcR had an expression pattern similar to that of PcRXR2. For PcRXR1, its expression remained at a certain high level, when there was a decrease of both PcEcR and PcRXR2. In situ hybridization showed that PcRXR2 was detected in the central nervous mass, while the ecdysteroid biosynthesis gene PcSpo was mainly expressed at the edge of the central nervous mass. RNAi-based silencing of PcRXR1 or PcRXR2 showed the same phenotype as in mites with that of silencing PcEcR. Furthermore, RNA-seq was used to mine the genes associated with the expression dynamics of PcRXR1 or PcRXR2, which revealed that the heterodimer of EcR-RXR2 in spider mites might be linked with the cell autophagy and tissue remodeling during apolysis, and RXR1 might be linked with new epicuticle and exocuticle secretion during ecdysis. Taken together, these results increase our understanding of the regulation mechanism of ecdysteroid signal pathway in spider mite development.

De novo RNA-Seq and Annotation of Sesquiterpenoid and Ecdysteroid Biosynthesis Genes and MicroRNAs in a Spider Mite Eotetranychus kankitus.

Eotetranychus kankitus is an important mite pest in citrus, but molecular data on the developmental processes of E. kankitus are lacking. The different development stages mix of E. kankitus was used to sequence for transcriptome and small RNAs to identify genes and predict miRNAs associated with sesquiterpenoid and ecdysteroid biosynthesis and signaling pathways. More than 36 million clean reads were assembled and 67,927 unigenes were generated. Of the unigenes, 19,300 were successfully annotated through annotation databases NR, SwissProt, COG, GO, KEGG, PFAM, and KOG. The transcripts were involved in sesquiterpenoid biosynthesis (11 genes) and ecdysteroid biosynthesis and signaling pathway (13 genes). Another, small RNA library was obtained and 31 conserved miRNAs were identified. Five most abundant miRNAs were Ek-miR-5735, Ek-miR-1, Ek-miR-263a, Ek-miR-184, and Ek-miR-8. The target genes related to sesquiterpenoid and ecdysteroid showed that 10 of the conserved miRNAs could potentially target the sesquiterpenoid and ecdysteroid pathway according to four-prediction software, sRNAT, miRanda, RNAhybrid, and Risearch2. Thus, the results of this study will provide bioinformatics information for further molecular studies of E. kankitus which may facilitate improved pest control strategies.

Baicalin attenuates LPS-induced alveolar type II epithelial cell A549 injury by attenuation of the FSTL1 signaling pathway via increasing miR-200b-3p expression.

In China, baicalin is the main active component of Scutellaria baicalensis, which has been used in the treatment of inflammation-related diseases, such as inflammation-induced acute lung injury. However, its specific mechanism remains unclear. This study examined the protective effect of baicalin on LPS-induced inflammation injury of alveolar epithelial cell line A549 and explored its protective mechanism. Compared with the LPS-induced group, the proliferation inhibition rates of alveolar type II epithelial cell line A549 intervened by different concentrations of baicalin decreased significantly, as did the levels of inflammatory factors IL-6, IL-1ß, prostaglandin 2 and TNF-α in the supernatant. The expression levels of inflammatory proteins inducible NO synthase (iNOS), NF-κB65, phosphorylated ERK (p-ERK1/2), and phosphorylated c-Jun N-terminal kinase (p-JNK1) significantly decreased, as did the protein expression of follistatin-like protein 1 (FSTL1). In contrast, expression of miR-200b-3p significantly increased in a dose-dependent manner. These results suggested that baicalin could significantly inhibit the expression of inflammation-related proteins and improve LPS-induced inflammatory injury in alveolar type II epithelial cells. The mechanism may be related to the inhibition of ERK/JNK inflammatory pathway activation by increasing the expression of miR-200b-3p. Thus, FSTL1 is the regulatory target of miR-200b-3p.

Folic acid-modified nonionic surfactant vesicles for gambogenic acid targeting: Preparation, characterization, and in vitro and in vivo evaluation.

The aim of present study was to develop folic acid (FA)-modified nonionic surfactant vesicles (NISVs, niosomes) as carrier systems for targeted delivery of gambogenic acid (GNA). The FA-GNA-NISVs exhibited a mean particle size of 180.77 ± 2.41 nm with a narrow poly dispersion index of 0.147 ± 0.08 determined by dynamic light scattering. Transmission electron microscopy also revealed that the FA-GNA-NISVs were spherical with double-layer structure. Entrapment efficiency (EE%) and zeta potential of the optimal FA-GNA-NISVs were 87.84 ± 1.06% and -37.33 ± 0.33 mV, respectively. Differential scanning calorimetry demonstrated that the GNA was in a molecular or amorphous state inside the FA-NISVs in vitro release profiles suggested that FA-GNA-NISVs could release GNA at a sustained manner, and less than 60% of GNA was released from the FA-NISVs within 12 hours of dialysis. in vivo pharmacokinetic results illustrated that FA-GNA-NISVs had considerably higher Cmax , area under curve (AUC0 - t ) and accumulation in lung. The cell proliferation study shown that the FA-GNA-NISVs significantly enhanced the in vitro cytotoxicity against A549 cells. Flow cytometry and fluorescence microscopy further demonstrated that the FA-GNA-NISVs increased apoptosis compared with nonmodified GNA-NISVs and free GNA. Moreover, FA-GNA-NISVs induced A549 cell apoptosis in a dose-dependent manner. In addition, cellular uptake assays showed a higher uptake of FA-GNA-NISVs than GNA-NISVs as well as free GNA. Taken together, it could be concluded that FA-GNA-NISVs were proposed as a novel targeting carriers for efficient delivering of GNA to cancers cells.