Integration of mRNA and miRNA analysis reveals the molecular mechanisms of sugar beet (Beta vulgaris L.) response to salt stress.

The continuous increase of saline-alkali areas worldwide has led to the emergence of saline-alkali conditions, which are the primary abiotic stress or hindering the growth of plants. Beet is among the main sources of sugar, and its yield and sugar content are notably affected by saline-alkali stress. Despite sugar beet being known as a salt-tolerant crop, there are few studies on the mechanisms underlying its salt tolerance, and previous studies have mainly delineated the crop's response to stress induced by NaCl. Recently, advancements in miRNA-mRNA network analysis have led to an increased understanding of how plants, including sugar beet, respond to stress. In this study, seedlings of beet variety "N98122" were grown in the laboratory using hydroponics culture and were exposed to salt stress at 40 days of growth. According to the phenotypic adaptation of the seedlings' leaves from a state of turgidity to wilting and then back to turgidity before and after exposure, 18 different time points were selected to collect samples for analysis. Subsequently, based on the data of real-time quantitative PCR (qRT-PCR) of salt-responsive genes, the samples collected at the 0, 2.5, 7.5, and 16 h time points were subjected to further analysis with experimental materials. Next, mRNA-seq data led to the identification of 8455 differentially expressed mRNAs (DEMs) under exposure to salt stress. In addition, miRNA-seq based investigation retrieved 3558 miRNAs under exposure to salt stress, encompassing 887 known miRNAs belonging to 783 families and 2,671 novel miRNAs. With the integrated analysis of miRNA-mRNA network, 57 miRNA-target gene pairs were obtained, consisting of 55 DEMIs and 57 DEMs. Afterwards, we determined the pivotal involvement of aldh2b7, thic, and δ-oat genes in the response of sugar beet to the effect of salt stress. Subsequently, we identified the miRNAs novel-m035-5p and novel-m0365-5p regulating the aldh gene and miRNA novel-m0979-3p regulating the thic gene. The findings of miRNA and mRNA expression were validated by qRT-PCR.

Du Huo Ji Sheng Tang inhibits Notch1 signaling and subsequent NLRP3 activation to alleviate cartilage degradation in KOA mice.

BACKGROUND: Knee osteoarthritis (KOA) has a complex pathological mechanism and is difficult to cure. The traditional medicine Du Huo Ji Sheng Tang (DHJST) has been used for the treatment of KOA for more than one thousand years, but its mechanism for treating KOA has not been revealed. In our previous study, we confirmed that DHJST inhibited the activation of NLRP3 signaling in rats and humans. In the current study, we aimed to determine how DHJST inhibits NLRP3 to alleviate knee cartilage damage. METHODS: Mice were injected with NLRP3 shRNA or Notch1-overexpressing adenovirus into the tail vein to construct systemic NLRP3 low-expressing or Notch1 high-expressing mice. Mice were injected with papain into the knee joint to replicate the KOA model. DHJST was used to treat KOA model mice with different backgrounds. The thickness of the right paw was measured to evaluate toe swelling. The pathohistological changes and the levels of IL-1ß, MMP2, NLRP3, Notch1, collagen 2, collagen 4, HES1, HEY1, and Caspase3 were detected by HE staining, ELISA, immunohistochemical staining, western blotting, or real-time qPCR. RESULTS: DHJST reduced tissue swelling and serum and knee cartilage IL-1ß levels, inhibited cartilage MMP2 expression, increased collagen 2 and collagen 4 levels, decreased Notch1 and NLRP3 positive expression rates in cartilage, and decreased HES1 and HEY1 mRNA levels in KOA model mice. In addition, NLRP3 interference decreased cartilage MMP2 expression and increased collagen 2 and collagen 4 levels without affecting the expression levels of notch1, HES1 and HEY1 mRNA levels in the synovium of KOA mice. In KOA mice with NLRP interference, DHJST further reduced tissue swelling and knee cartilage damage in mice. Finally, Notch1-overexpressing mice not only showed more severe tissue swelling and knee cartilage degradation but also abolished the therapeutic effect of DHJST on KOA mice. Importantly, the inhibitory effects of DHJST on the mRNA expression of NLRP3, Caspase3 and IL-1ß in the knee joint of KOA mice were completely limited after Notch1 overexpression. CONCLUSION: DHJST significantly reduced inflammation and cartilage degradation in KOA mice by inhibiting Ntoch1 signaling and its subsequent NLRP3 activation in the knee joint.

Therapeutic efficacy of traditional Chinese exercises and massage treatment on lumbar instability: a randomized controlled trial.

OBJECTIVE: To determine the effect of traditional Chinese exercises combined with traditional Chinese massage on lumbar instability. METHODS: Seventy patients with lumbar instability were randomly divided into experimental group and control group. The experimental group was treated with traditional Chinese exercises combined with traditional Chinese massage, while the control group was only treated with traditional Chinese exercises. The Japanese Orthopaedic Association (JOA) scores, Oswestry Disability Index (ODI) and dynamic imaging indexes were used to assess treatment outcomes. RESULTS: After 8 weeks of treatment, the JOA scores of both groups were significantly higher than those before treatment (P < 0.05), while ODI was significantly lower (P < 0.05). In addition, compared with the control group, the JOA score of the experimental group was significantly increased (P < 0.05), and the ODI was significantly decreased (P < 0.05). Besides, the dynamic imaging indexes of patients with lumbar instability significantly changed after treatment. The lumbar vertebral laxity was significantly decreased (P < 0.05) than those before treatment. CONCLUSION: The traditional Chinese exercises and massage may improve the symptoms of lumbar instability, and traditional Chinese exercises combined with massage treatment may be better than exercises alone.

Massage manipulation vs. low back muscle exercise for lumbar intervertebral instability: A preliminary randomized clinical trial.

This is a preliminary randomized clinical trial on patients conducted at Wuxi Hospital Affiliated with Nanjing University of Chinese Medicine from September 2015 to December 2016. The patients with intervertebral instability were randomized 1:1 for massage (20 min/day for 6 days) or exercise (3 sessions/day for 15 days). Japanese Orthopaedic Association (JOA) score, Oswestry disability score, and quantitative fluoroscopy (QF) were performed before and after the treatment and at 1 and 3 months thereafter. Improvement rates were noted to be 86.7% and 40.0% in the massage and exercise groups, respectively. Massage group showed significant changes in the JOA and Oswestry disability scores (p < 0.001 and p = 0.002), while the exercise group did not show any significant change (p > 0.05). Changes in the JOA and Oswestry disability scores were more important in the massage group (p < 0.05). All dynamic imaging parameters were improved in the massage group (all p < 0.05) but not in the exercise group (all p>0.05). These results suggest that the massage manipulation could be an appropriate way to treat intervertebral instability.