RESUMO
The calmodulin-binding protein 60 (CBP60) family is a gene family unique to plants, and its members play a crucial role in plant defense responses to pathogens and growth and development. Considering that cotton is the primary source of natural cotton textile fiber, the functional study of its CBP60 gene family members is critical. In this research, we successfully identified 162 CBP60 members from the genomes of 21 species. Of these, 72 members were found in four cotton species, divided into four clades. To understand the function of GhCBP60B in cotton in depth, we conducted a detailed analysis of its sequence, structure, cis-acting elements, and expression patterns. Research results show that GhCBP60B is located in the nucleus and plays a crucial role in cotton growth and development and response to salt and drought stress. After using VIGS (virus-induced gene silencing) technology to conduct gene silencing experiments, we found that the plants silenced by GhCBP60B showed dwarf plants and shortened stem nodes, and the expression of related immune genes also changed. In further abiotic stress treatment experiments, we found that GhCBP60B-silenced plants were more sensitive to drought and salt stress, and their POD (peroxidase) activity was also significantly reduced. These results imply the vital role of GhCBP60B in cotton, especially in regulating plant responses to drought and salt stress. This study systematically analyzed CBP60 gene family members through bioinformatics methods and explored in depth the biological function of GhCBP60B in cotton. These research results lay a solid foundation for the future use of the GhCBP60B gene to improve cotton plant type and its drought and salt resistance.
Assuntos
Proteínas de Ligação a Calmodulina , Regulação da Expressão Gênica de Plantas , Gossypium , Estresse Fisiológico , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/metabolismo , Secas , Genoma de Planta , Gossypium/genética , Gossypium/metabolismo , Família Multigênica , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genéticaRESUMO
ObjectiveTo investigate whether phosphodiesterase (PDE) 5 inhibitors sildenafil (SIL) or LW1646 prevented renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO). MethodsMale C57BL/6 mice were randomly divided into four groups (n =6), namely the Sham group, 7UUO group, 7UUO+SIL group and 7UUO+LW1646 group. Sildenafil (SIL) or LW1646, or vehicle was administered 1 hour before surgery, and the mice were continuously treated once daily (i. g., 50 mg/kg) for 7 days. The obstructed kidneys were harvested on day 7. Hematoxylin-eosin (HE) and Masson’s staining was used to examine renal histology. Immunoblotting and RT-qPCR were used to detect the expression levels of protein and mRNA for fibrosis, apoptosis, endoplasmic reticulum (ER) stress, autophagy, and pro-fibrotic factors. Human proximal tubule epithelial cells (HK-2) were treated with TGF-β1 for 48 hours or tunicamycin for 24 hours, respectively, to evaluate whether cyclic guanosine monophosphate (cGMP) or PDE5 inhibitors prevents ER stress and pro-fibrotic responses. ResultsAt the 7th days after UUO, the body weight of the mice showed a significant decrease (P< 0.000 1) compared with that in the sham group. The obstructed kidneys showed a significant tubular dilation and interstitial inflammation. The levels of protein and mRNA expression in apoptosis, ER stress, autophagy-related protein and pro-fibrotic factors were also markedly increased in UUO mice (P <0.05). In contrast, SIL or LW1646 treatment was associated with attenuated tubular dilation, infiltration of inflammatory cells and collagen content in the obstructed kidney of the mice. The protein and mRNA expression levels of renal TGF-β1 were markedly decreased, and the protein expression levels of apoptosis, endoplasmic reticulum stress, and autophagy markers were also significantly downregulated by PDE5 inhibitors. In HK-2 cells, TGF-β1 induced increased expression levels of fibronectin and BiP, which was at least partially reversed by cGMP, a product of PDE inhibition. Additionally, PDE5 inhibitors were found to modulate aberrant levels of autophagy and apoptosis. ConclusionIn conclusion, PDE5 inhibitors, in particular, LW1646, can alleviate the progression of fibrosis by improving ER stress, apoptosis and autophagy as well as downregulating protein and mRNA expression of TGF-β1.
RESUMO
ObjectiveTo investigate the role of bile acid receptor TGR5 activation in renal fibrosis induced by unilateral ischemia reperfusion injury and contralateral nephrectomy (uIRIx) model. MethodsIn vivo: C57BL/6J mice were randomly divided into Sham group, uIRIx group and uIRIx+ lithcholic acid (LCA) group with 6 mice in each group. Kidney fibrosis was induced by uIRIx model, kidney function was evaluated by blood and urine biochemical indexes, and the degree of kidney injury was evaluated by HE staining. Masson staining and immunohistochemistry were used to evaluate the degree of renal fibrosis, and Western Blotting was used to detect the expression of related index proteins of renal cortical fibrosis. Sham group and uIRIx group were set in TGR5+/+ mice and TGR5-/- mice respectively, with 6 mice in each group. The degree of renal fibrosis in each group was detected by Western Blotting. In vitro: TGF-β1 was administered to induce pro-fibrosis response in human renal tubular epithelial cell line (HK2 cells), LCA was used for drug intervention, cytoskeleton was labeled with phalloidin-FITC staining and the expression of fibrosis related indicator protein in HK2 cells was detected by Western Blotting. ResultsIn vivo: Compared with the Sham group, plasma creatinine level (P=0.007) and urinary albumin/creatinine ratio (P=0.041) in uIRIx group were significantly increased, renal cortical protein TGR5 expression (P=0.002) was decreased, Fibronectin expression (P=0.020) and COL1A1 expression (P<0.001) were increased. At the same time, the kidney structure was damaged and collagen deposition was aggravated. LCA intervention effectively improved the kidney function and alleviated the degree of kidney injury and fibrosis. TGR5 gene knockout increased uIRIx-induced Fibronectin expression (P<0.001) and COL1A1 expression (P=0.001) compared with TGR5+/+ mice. In vitro: TGF-β1 induced morphological changes of HK2 cells, cytoskeletal depolymerization and recombination, and promoted the up-regulation of fibrosis index protein. LCA effectively inhibited the morphological changes and skeletal depolymerization induced by TGF-β1, and down-regulated the expression of fibrosis related indicator proteins. ConclusionsLCA alleviated renal fibrosis induced by uIRIx model, and knockout of TGR5 gene aggravated uIRIx induced renal fibrosis; In HK2 cells, LCA alleviated fibrogenic reaction induced by TGF-β1. This indicates that activation of TGR5 alleviates renal fibrosis induced by uIRIx.
RESUMO
The purpose of this study was to investigate how Zuogui pills from the Kidney-tonifying and Nourishing Yin formula, in combination with the gonadotrophin-releasing hormone antagonist cetrorelix, affected the ovarian local oxidative stress response in decreasing ovarian reserve (DOR) mice. All animal experiments were carried out in accordance with the guidelines and standards established by Jinan University's Experimental Animal Management Committee. Cyclophosphamide (CTX)-treated DOR mice were given Zuogui pills, cetrorelix, or a combination of the two drugs intragastrically. After treatment, there were changes in the estrous cycle, serum sex hormone levels, oxidative stress-related indexes, growth biochemical factor levels, and SIRT1/P53/P21 expression. In comparison to the model group, the Zuogui pills and the cetrorelix+Zuogui pills group had significantly prolonged estrous periods and shortened interestrous periods, and the cetrorelix+Zuogui pills group had a significantly shortened cycle length. Follicle-stimulating hormone (FSH) decreased and estradiol (E2) increased in all treatment groups compared to the model group, oxidative stress indexes nitric oxide synthase (NOS), nitric oxide (NO), and reactive oxygen species (ROS) decreased, growth biochemical factors brain derived neurotrophic factor (BDNF) and growth differentiation factor 9 (GDF-9) concentrations increased significantly, and leukemia inhibitory factor (LIF) showed no significant change. SIRT1/P53/P21 immunohistochemical results revealed that, when compared to the model group, the expression of SIRT1 increased while the expression of P53 and P21 proteins decreased in all treatment groups, with the cetrorelix+Zuogui pills group having the largest decrease, with significant differences in all indicators. We conclude that cetrorelix combined with Zuogui pills for kidney nourishing and Yin recipe improved the oxidative stress response in the follicle by regulating the SIRT1/P53/P21 pathway, reducing peroxide product production, protecting ovarian function, and regulating ovarian hormone secretion, and its efficacy is superior to that of cetrorelix or Zuogui pills alone.
RESUMO
OBJECTIVE@#To investigate the incidence of autism spectrum disorder (ASD)-like symptoms in the population with intellectual disability (ID).@*METHODS@#The students with ASD or ID, aged 6-18 years, who studied in a special school in Shanghai from January to June, 2017, as well as the typically developing (TD) population of the same age, who studied in a general school in Shanghai during the same period, were enrolled. Social Responsiveness Scale (SRS) was completed by their parents or other guardians, and the ASD-like symptoms were evaluated.@*RESULTS@#A total of 69 subjects with ASD, 74 subjects with ID and 177 TD subjects were enrolled. The ID group had a significantly higher SRS-positive rate than the TD group (47.3% vs 1.7%; P0.05), and there was no significant correlation between SRS score and IQ (P>0.05).@*CONCLUSIONS@#The ID population aged 6-18 years has more ASD-like symptoms than the general population, and ASD screening and intervention should be performed for the ID population as early as possible.
Assuntos
Adolescente , Criança , Humanos , Transtorno do Espectro Autista , China , Deficiência Intelectual , Pais , Comportamento SocialRESUMO
<p><b>INTRODUCTION</b>The use of an additional biopsy from the gastric body may help improve the detection of Helicobacter pylori during endoscopy. This study aimed to determine whether such an additional biopsy is necessary in routine rapid urease test (RUT), and whether acid suppression and antibiotic therapy affect RUT results.</p><p><b>METHODS</b>Patients recruited had two gastric mucosal biopsies taken - one from the gastric antrum and the other from the gastric body. Each biopsy was placed into separate RUT kits. Information on previous or current use of proton-pump inhibitors, H2 receptor antagonist, bismuth and antibiotics was obtained. Patients on any of those drugs one week prior to endoscopy were considered to have a positive drug history (PDH).</p><p><b>RESULTS</b>Of the 400 patients recruited, 311 had negative RUTs and 89 had at least one positive RUT. Between the PDH and negative drug history (NDH) groups, there was a significant difference in the distribution of the location of the biopsies that yielded positive RUTs (p = 0.023). The NDH group had a higher proportion of patients who had positive RUTs for both locations, whereas the PDH group had a higher proportion of patients who had positive RUTs for only one location.</p><p><b>CONCLUSION</b>As RUT results are significantly affected by the use of acid suppression and antibiotic therapies, biopsies for RUT should be taken from both the gastric antrum and body to minimise false negative results.</p>