Rice is the world's largest food crop, and its yield and quality are directly related to food security and human health. Grain size, as one of the important factors determining the rice yield, has been widely concerned by breeders and researchers for a long time. To decipher the regulatory mechanism of rice grain size, we obtained a multi-tiller, dwarf, and small-grain mutant htd1 by ethyl methanesulfonate (EMS) mutation from the Japonica rice cultivar 'Zhonghua 11' ('ZH11'). Genetic analysis indicated that the phenotype of htd1 was controlled by a single recessive gene. Using the mutation site map (Mutmap) method, we identified the candidate gene OsHTD1, which encoded a carotenoid cleavage dioxygenase involved in the biosynthesis of strigolactone (SL). The SL content in htd1 was significantly lower than that in 'ZH11'. Cytological analysis showed that the grain size of the mutant decreased due to the reductions in the length and width of glume cells. The function of htd1 was further verified by the CRISPR/cas9 gene editing technology. The plants with the gene knockout exhibited similar grain size to the mutant. In addition, gene expression analysis showed that the expression levels of multiple grain size-related genes in the mutant changed significantly, suggesting that HTD1 may interact with other genes regulating grain size. This study provides a new theoretical basis for research on the regulatory mechanism of rice grain size and potential genetic resources for breeding the rice cultivars with high yields.
Oryza/growth & development* ; Mutation ; Edible Grain/growth & development* ; Alleles ; Plant Proteins/genetics* ; Dioxygenases/genetics* ; Lactones/metabolism* ; Gene Expression Regulation, Plant ; Genes, Plant ; Gene Editing ; CRISPR-Cas Systems ; Phenotype

Objective To observe the effect of Shenqi Chongcao (SQCC) Formula on the ASS1/src/STAT3 signaling pathway in a rat model of lung fibrosis and explore its therapeutic mechanism. Methods A total of 120 male SD rats were divided equally into 5 groups, including a blank control group with saline treatment and 4 groups of rat models of idiopathic pulmonary fibrosis induced by intratracheal instillation of bleomycin. One day after modeling, the rat models were treated with daily gavage of 10 mL/kg saline, SQCC decoction (0.423 g/kg), pirfenidone (10 mL/kg), or intraperitoneal injection of arginine deiminase (ADI; 2.25 mg/kg, every 3 days) for 28 days. After the treatments, the lung tissues of the rats were collected for calculating the lung/body weight ratio, observing histopathology using HE and Masson staining, and analyzing the inflammatory cells in BALF using Giemsa staining. Serum chemokine ligand 2 (CCL2) and transforming growth factor-β1 (TGF-β1) levels were measured with ELISA. The protein expressions of src, p-srcTry529, STAT3, and p-STAT3Try705 and the mRNA expressions of ASS1, src and STAT3 in the lung tissues were detected using Western blotting and RT-qPCR. Results The neutrophil, macrophage and lymphocyte counts and serum levels of CCL2 and TGF-β1 were significantly lower in SQCC, pirfenidone and ADI treatment groups than in the model group at each time point of measurement (P<0.05). P-srcTry529 and p-STAT3Try705 protein expression levels and ASS1, src, and STAT3 mRNA in the lung tissues were also significantly lower in the 3 treatment groups than in the model group (P<0.05). Conclusion SQCC Formula can alleviate lung fibrosis in rats possibly by activating the ASS1/src/STAT3 signaling pathway in the lung tissues.

Objective To observe the effect of Shenqi Chongcao (SQCC) Formula on the ASS1/src/STAT3 signaling pathway in a rat model of lung fibrosis and explore its therapeutic mechanism. Methods A total of 120 male SD rats were divided equally into 5 groups, including a blank control group with saline treatment and 4 groups of rat models of idiopathic pulmonary fibrosis induced by intratracheal instillation of bleomycin. One day after modeling, the rat models were treated with daily gavage of 10 mL/kg saline, SQCC decoction (0.423 g/kg), pirfenidone (10 mL/kg), or intraperitoneal injection of arginine deiminase (ADI; 2.25 mg/kg, every 3 days) for 28 days. After the treatments, the lung tissues of the rats were collected for calculating the lung/body weight ratio, observing histopathology using HE and Masson staining, and analyzing the inflammatory cells in BALF using Giemsa staining. Serum chemokine ligand 2 (CCL2) and transforming growth factor-β1 (TGF-β1) levels were measured with ELISA. The protein expressions of src, p-srcTry529, STAT3, and p-STAT3Try705 and the mRNA expressions of ASS1, src and STAT3 in the lung tissues were detected using Western blotting and RT-qPCR. Results The neutrophil, macrophage and lymphocyte counts and serum levels of CCL2 and TGF-β1 were significantly lower in SQCC, pirfenidone and ADI treatment groups than in the model group at each time point of measurement (P<0.05). P-srcTry529 and p-STAT3Try705 protein expression levels and ASS1, src, and STAT3 mRNA in the lung tissues were also significantly lower in the 3 treatment groups than in the model group (P<0.05). Conclusion SQCC Formula can alleviate lung fibrosis in rats possibly by activating the ASS1/src/STAT3 signaling pathway in the lung tissues.

Excavating the quantitative trait locus (QTL) associated with rice cooking quality, analyzing candidate genes, and improving cooking quality-associated traits of rice varieties by genetic breeding can effectively improve the taste of rice. In this study, we used the indica rice HZ, the japonica rice Nekken2 and 120 recombinant inbred lines (RILs) populations constructed from them as experimental materials to measure the gelatinization temperature (GT), gel consistency (GC) and amylose content (AC) of rice at the maturity stage. We combined the high-density genetic map for QTL mapping. A total of 26 QTLs associated with rice cooking quality (1 QTL associated with GT, 13 QTLs associated with GC, and 12 QTLs associated with AC) were detected, among which the highest likelihood of odd (LOD) value reached 30.24. The expression levels of candidate genes in the localization interval were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), and it was found that the expression levels of six genes were significantly different from that in parents. It was speculated that the high expression of LOC_Os04g20270 and LOC_Os11g40100 may greatly increase the GC of rice, while the high expression of LOC_Os01g04920 and LOC_Os02g17500 and the low expression of LOC_Os03g02650 and LOC_Os05g25840 may reduce the AC. The results lay a molecular foundation for the cultivation of new high-quality rice varieties, and provide important genetic resources for revealing the molecular regulation mechanism of rice cooking quality.
Quantitative Trait Loci ; Oryza/genetics* ; Plant Breeding ; Cooking ; Genetic Association Studies

Objective:To explore the early predictive value of urinary nephrin in acute kidney injury (AKI) for critically ill neonates.Methods:A prospective study was conducted to neonates who were admitted to Neonatal Intensive Care Unit (NICU) Children′s Hospital of Soochow University, from July to October 2016.According to whether AKI occurred during the NICU′s hospitalization, neonates were divided into AKI group and non-AKI group.Urinary nephrin levels were detected at the first 24 h of NICU, and the score for neonatal acute physiology (SNAP) was assessed within 24 hours of NICU.Multivariate linear analyses were applied to analyze potential variables that were asso-ciated with urinary nephrin level.Multivariate Logistic regression analysis was adopted to evaluate the relationship between urinary nephrin and AKI after adjusting for confounding factors.A receiver operating characteristic(ROC) curve and the area under the ROC curve (AUC) were calculated to assess the early predictive value of urinary nephrin for neonatal AKI. Results:Among the 156 neonates enrolled in the study, 16 cases(10.2%) developed AKI.The median of urinary nephrin, urinary albumin and SNAP scores were 0.27 μg/mg uCr, 0.48 g/g uCr and 9 scores with AKI group, while the median of urinary nephrin, urinary albumin and SNAP scores were 0.16 μg/mg uCr, 0.16 g/g uCr and 7 scores with non-AKI group.When compared with non-AKI neonates, urinary nephrin ( Z=-3.201, P=0.001), urinary albumin ( Z=-2.652, P=0.008) and SNAP score ( Z=-2.611, P=0.009) were significantly higher in AKI neonates.Multiple linear regression analysis proved that urinary nephrin levels were significantly correlated with urinary albumin ( B=0.488, SE=0.117, P<0.001). Multivariate Logistic regression analysis revealed that urinary nephrin remained significantly associated with AKI ( P=0.018) after adjusting for confounding factors, including gestational age, birth weight, gender, SNAP score, mechanical ventilation and apnea.Urinary nephrin achieved AUC of 0.746 (95% CI: 0.606-0.886, P=0.001). Conclusions:As a biomarker of glomerular injury, urinary nephrin is significantly related to the occurrence of AKI and has early predictive value for AKI in critically ill neonates.

Objective To investigate the expression of FAM19A4 gene promotor methylation in different cervical lesions and its diagnostic value. Methods 31 cervical cancers cases ,22 HSILs and 23 normal cervical tissues of formalin-fixed and parrffin-embedded specimen diagnosed by pathology were selected. Taqman probe-based quantitative PCR(qPCR)was used to detect the differences of FAM19A4 methylation in different cervical lesions, and then corresponding analyses were made. Results Statistical differences of FAM19A4 methylationrates were observed in cervical caner ,HSIL and normalcervix,respectively96.8%(30/31),72.7%(16/22)and 8.7%(2/23) (P < 0.05 ).FAM19A4 methylation rates increased with severity of cervical lesion (P < 0.05).The methylation rates of FAM19A4 were not statistically different in different clinicopathological characteristics of cervical cancer (P>0.05). Conclusions FAM19A4 gene promoter methylation is probably a specific biomarker of cervical cancer,andmay play a role in the development and progress of cervical cancer,but may not participate in the invasion and metastasis.

Objectives To investigate the association of fluid overload (FO) with the development and mortality of acute kidney injury (AKI) and to evaluate the predictive value of FO in mortality of critically ill children. Method A prospective study was conducted among critically ill children who were admitted to the children's intensive care unit (PICU). FO levels were assessed during the course of the disease and PRISM Ⅲ scores were evaluated within 24 hours of admission. Binary logistic regression analyses were conducted to evaluate the association of FO with the development and mortality of AKI after adjusting for confounding factors. The area under the receiver operating characteristic curve (AUC) was calculated to assess the predictive value of FO for mortality. Results In 362 children included, there were 26 children (7.18%) having average FO≥5%, and AKI in 24 children (6.63%) and 18 children (5.0%) died. The mean FO (OR=1.26, 95%CI: 1.10～1.43, P=0.001) and the maximum FO (OR=1.12, 95%CI: 1.02～1.23, P=0.018) were significantly correlated with the development of AKI in critically ill children within 7 days of admission to PICU. However, after adjusting for age and PRISM Ⅲ, both factors had no association with AKI (all P>0.05). After adjusting for the potential confounders such as AKI and the severity of disease, the average FO was significantly associated with mortality (AOR=1.34, 95%CI: 1.12～1.60, P=0.002). The AUC of mean FO that predicted mortality risk was 0.801 (P<0.001). Conclusion Fluid overload is associated with the development and the prognosis of AKI in critically ill children, and has important predictive value for mortality.