N6-methyladenosine mRNA methylation positively regulated the response of poplar to salt stress.

As the most abundant form of methylation modification in messenger RNA (mRNA), the distribution of N6-methyladenosine (m6A) has been preliminarily revealed in herbaceous plants under salt stress, but its function and mechanism in woody plants were still unknown. Here, we showed that global m6A levels increased during poplar response to salt stress. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) revealed that m6A significantly enriched in the coding sequence region and 3'-untranslated regions in poplar, by recognising the conserved motifs, AGACU, GGACA and UGUAG. A large number of differential m6A transcripts have been identified, and some have been proved involving in salt response and plant growth and development. Further combined analysis of MeRIP-seq and RNA-seq revealed that the m6A hypermethylated and enrich in the CDS region preferred to positively regulate expression abundance. Writer inhibitor, 3-deazaneplanocin A treatment increased the sensitivity of poplar to salt stress by reducing mRNA stability to regulate the expression of salt-responsive transcripts PagMYB48, PagGT2, PagNAC2, PagGPX8 and PagARF2. Furthermore, we verified that the methyltransferase PagFIP37 plays a positively role in the response of poplar to salt stress, overexpressed lines have stronger salt tolerance, while RNAi lines were more sensitive to salt, which relied on regulating mRNA stability in an m6A manner of salt-responsive transcripts PagMYB48, PagGT2, PagNAC2, PagGPX8 and PagARF2. Collectively, these results revealed the regulatory role of m6A methylation in poplar response to salt stress, and revealed the importance and mechanism of m6A methylation in the response of woody plants to salt stress for the first time.

A randomized, parallel control and multicenter clinical trial of evidence-based traditional Chinese medicine massage treatment VS External Diclofenac Diethylamine Emulgel for the treatment of knee osteoarthritis.

BACKGROUND: Both massage and topically administered NSAIDs are safe and effective treatments for knee osteoarthritis (KOA); however, different massage technique sects in China caused assessment difficulties for the treatment of KOA. In order to standardize the massage techniques and procedures, we organized multi-disciplinary experts in China to acquire an evidence-based traditional Chinese medicine massage treatment of knee osteoarthritis. The purposes of this study will be to provide clinicians a complementary and alternative therapy for patients and to evaluate the efficacy and safety of evidence-based traditional Chinese medicine massage treatment of KOA compared to External Diclofenac Diethylamine Emulgel. METHODS AND DESIGN: A randomized controlled trial in which 300 participants diagnosed with KOA will be recruited and randomly allocated to either the experimental group or the control group in a ratio of 2:1. Two hundred participants will receive evidence-based traditional Chinese medicine massage 2 sessions per week for 10 weeks as the experimental group, and 100 participants will receive External Diclofenac Diethylamine Emulgel 3-4 times per day for 10 weeks as the control group. The patients in the two groups will receive follow-up at two time points at 5 weeks and 10 weeks from the beginning of treatment, respectively. The MRI scans and X-ray will be performed at baseline and at the end of the intervention. The primary outcome will be the changes in the Western Ontario and McMaster Osteoarthritis Index (WOMAC). Secondary outcomes will be measured by the PRO scale for knee osteoarthritis based on the concept of traditional Chinese medicine (Chinese scale for knee osteoarthritis (CSKO)), X-ray evaluation, and MRI scan evaluation. The data of WOMAC and CSKO will be analyzed at the baseline, 5 weeks, and 10 weeks from the beginning of treatment. The data from MRI scans and X-rays will be analyzed at baseline and at the end of the intervention. The significance level sets as 5%. The safety of interventions will be evaluated after each treatment session. DISCUSSION: This study will provide clinicians with much-needed knowledge for the treatment of KOA through a controlled trial. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR1800014400 . Registered on 10 January 2018.

Reduced number of IFN-γ producing cells in peripheral blood is a biomarker for patients with renal cell carcinoma.

Renal cell cancer (RCC) is the most lethal of all the common urologic cancers and constitutes 2.2% of all malignancy diagnoses. The incidence of RCC has been steadily increasing in recent decades. The classic risk factors of RCC include smoking, hypertension, obesity, genetics, and genetic mutations. Recent studies also revealed that RCC was an immunogenic tumor and affected by host immune status. Among the pan-cance, RCC presented with the highest degree of immune infiltration, indicating RCC patients might benefit from immunotherapy. A new immune classification of RCC has been developed by Su et al. based on tumor-infiltrating lymphocytes to guide clinical practice. However, these studies mainly focus on biomarkers derived from tumor microenvironment (TME), the biomarkers based on peripheral blood samples to RCC have rarely been described. We collected peripheral blood samples from RCC patients and their matched healthy controls and detected the number of IL-2 and IFN-γ producing cells by implementing an enzyme-linked immunospot (ELISPOT) assay. This is the first study to report blood-based immune biomarkers for RCC using an ELISPOT assay. Our results suggested the frequency of IFN-γ producing cells but not IL-2 producing cells was associated with RCC risk. These findings warrant further validation in larger prospective studies.

Internet addiction and suicidal behavior among vocational high school students in Hunan Province, China: A moderated mediation model.

Background: Vocational education is an important part of high school education in China. However, there is little research on high school students' mental health. This study aimed to investigate the prevalence of suicidal behavior (SB) among this population and the mediating role of insomnia, depression, anxiety, and stress in the relationship between Internet addiction (IA) and SB using a structural equation model. Methods: A cross-sectional questionnaire survey was conducted among several vocational high school students in Hunan Province, and 7,968 valid questionnaires were obtained. General demographic data and data from the Dual-Mode Self-Control Scale, Athens Insomnia Scale, Depression Anxiety Stress scale-21, and Revised Chen Internet Addiction Scale were collected. A structural equation model was used to explore the different pathways from IA to SB. Results: Among the participants, 37.7, 15.7, and 21.8% reported suicidal ideation, plans, and attempts, respectively. The structural equation model confirmed that IA was indirectly related to SB and was mediated by insomnia and/or depression, anxiety, and stress. Limitations: First, we only recruited students from vocational schools in Hunan Province, therefore, the sample may not represent the entire population of vocational students in China. Second, self-report scales were used in this study, and clinical diagnosis required professional interviews. Third, since this study had a cross-sectional design, the causal relationship between the variables could not be determined. Conclusions: The prevalence of SB among vocational high school students in China was significantly high. The prevention of SB related to IA can be attributed to the improvement of insomnia and emotional problems.

Comparison of ultrasound biomicroscopic changes after glaucoma triple procedure and trabeculectomy in eyes with primary angle closure glaucoma.

PURPOSE: To evaluate ultrasound biomicroscopy changes in the anterior chamber angle (ACA) configuration after glaucoma triple procedure (GTP), compared with those seen after trabeculectomy, in eyes with primary angle closure glaucoma (PACG). METHODS: Twenty-nine PACG eyes that underwent GTP were compared with 27 eyes that underwent trabeculectomy only, using ultrasound biomicroscopy. Anterior segment parameters such as anterior chamber depth, angle opening distance (AOD500), trabecular ciliary process distance, trabecular iris angle, iris ciliary process distance, iris thickness along the trabecular ciliary process distance line (ID1), scleral ciliary process angle, scleral iris angle, and angle recess area were measured preoperatively and at 3 months postoperatively; these parameters were then compared between the procedures. RESULTS: Among the anterior segment parameters, postoperative increases in the anterior chamber depth, AOD500, and angle recess area were statistically significant in the eyes that underwent trabeculectomy. For the eyes that underwent GTP, all parameters except the iris ciliary process distance and ID1 were significantly increased after the operation. CONCLUSIONS: For the eyes that underwent GTP, the ciliary body process was posteriorly positioned and the ACA was significantly increased after the operation, as compared with the eyes that underwent trabeculectomy. The posterior shift of the ciliary process only after GTP was indirect evidence that the lens plays an important role in ACA configuration in eyes with PACG.

Ultrasound biomicroscopic changes after laser iridotomy or trabeculectomy in angle-closure glaucoma.

This study was performed to demonstrate the ultrasound, biomicroscopic and dimensional changes of angle structure after laser iridotomy (LI) and primary trabeculectomy (PT) in primary angle-closure glaucoma (PACG). Angle-opening distance at a point 500 m from the scleral spur (AOD500), trabecular-iris angle (theta1), trabecular ciliary process distance (TCPD), ciliary process-iris angle (CPI), iris thickness (ID1, ID3), length of iris-lens contact distance (ILCD) and anterior chamber depth (ACD) were assessed before and after each procedure. Thirteen patients with LI and 16 with PT were prospectively enrolled. There were statistically significant increases in AOD500, theta1, and ILCD in both groups. CPI was decreased in both groups. ACD, TCPD, and iris thickness were not changed significantly. The changes in angle configuration after LI or PT may result more from alterations in aqueous pressure gradients across the iris and the changes of configuration were greater in the iris roots without rotation of ciliary body. However, we didn't find any significant differences in the changes of parameters between the two procedures.

Effect of human RAD50 gene therapy on glaucoma filtering surgery in rabbit eye.

PURPOSE: To investigate the antiproliferative effects of the subconjunctival injection of human RAD50 (hRAD50) on fibroblasts after glaucoma filtering surgery. METHODS: After glaucoma filtering surgery in normal rabbit eye, the subconjunctival injection of hRAD50 was performed. Morphologic changes in the subconjunctival area of hRAD50-treated eyes were compared with those of mitomycin C (MMC)-treated and control eyes using light and electron microscopy. Results. Two weeks after hRAD50 treatment (2 microg), the conjunctival epithelium increased in thickness and had many tonofilaments, but the basal lamina was intact. The subconjunctival fibroblasts exhibited a granular endoplasmic reticulum without distension. Most of the collagen bundles and fibers around the fibroblasts disappeared, and apoptotic cells with many fragmented nuclei and condensed chromatin were observed. Four weeks after hRAD50 treatment, the findings were similar with those of 2 weeks except for a slight increase in the number of collagen bundles and fibers and the appearance of macrophages. In MMC-treated eyes, the conjunctival epithelium also increased in thickness and had many tonofilaments. However, intercellular spaces were widened and the basal lamina was interrupted in some areas. Most of the collagen bundles and fibers were shortened, and apoptotic cells were also observed. CONCLUSIONS: These results demonstrate that the histologic antiproliferative effects of local hRAD50 on the conjunctival fibroblasts are similar to those of MMC, but without damage to the basal lamina of the conjunctival epithelium, and suggest that hRAD50 may be useful as a possible antifibroblastic agent for glaucoma filtering surgery. However, further investigations are needed to test for possible systemic complications, which so far have not been reported.

Ultrasound biomicroscopic dimensions of the anterior chamber in angle-closure glaucoma patients.

In order to evaluate the morphologic types of appositional angle-closure glaucoma, biometric measurements were made in angle-closure glaucoma patients using Ultrasound biomicroscopy (UBM). Twenty-six patients with primary angle-closure glaucoma and 21 cataract patients with as a control group were examined. The angle-closure glaucomatous eyes were classified as type B in which the angle closure started at the bottom of the angle and type S in which the angle closure occurred in the vicinity of Schwalbe's line. The trabecular-ciliary process distance (TCPD, type B; 873.20+/-86.77 microm, type S; 832.52+/-82.96 microm, control; 1233.50+/-73.01 microm, p = 0.000) and the angle opening distance (AOD500, type B; 89.75+/-63.27 microm, type S; 88.85+/-72.95 microm, control; 304.40+/-104.30 microm, p = 0.000) were significantly shorter in patients with angle closure vice control group. No significant difference were noted in the three groups of patients in regards to iris thickness or ciliary process-iris angle. In this study, we have demonstrated that there are two types of appositional angle-closure and have shown the forward rotation of the ciliary process without changes of the ciliary process-iris angle in cases of angle-closure glaucoma.

Postnatal expression and distribution of Refsum disease gene associated protein in the rat retina and visual cortex: effect of binocular visual deprivation.

Previously, phytanoyl-CoA alpha-hydroxylase-associated protein 1 (PAHX-AP1) was isolated as a novel neuron-specific protein to interact with Refsum disease (RfD) gene PAHX. Its expression in the brain increased after eyelid opening, and the elevated level was maintained through adulthood. In this report, to verify the hypothesis that light could trigger this increase, we have examined the developmental distribution pattern of PAHX-AP1 in rat retina and visual cortex, and changes of its expression by binocular deprivation. Northern blot analyses demonstrated PAHX-AP1 expression reached its highest level in the visual cortex and eyeball at 4 weeks after birth, and these levels were maintained through adult life. Two weeks after visual deprivation, its expression in the eyeball and visual cortex decreased compared with the control. In situ hybridization analyses of the retina showed that PAHX-AP1 expression was limited to the ganglionic cell layer at 10 days after birth, but expressed in the inner nuclear cell layer and extended to the outer nuclear cell layer at 2 and 3 weeks after birth, respectively. Two weeks after visual deprivation, however, it decreased in the ganglionic and inner nuclear cell layer, and disappeared in the rod and cone cell layers. In the visual cortex, strong signals of PAHX-AP1 were detected in layers IV and VI, and II-VI at 10 days and 2 weeks after birth, respectively. Its expression decreased after 2 weeks of visual deprivation. These results indicate that visual stimulation is essential for the maintenance of PAHX-AP1 expressions in the retina, especially in the rod and cone cell layers, and visual cortex, and suggest that PAHX-AP1 may be involved in the developmental regulation of the photoreceptor's function.