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Internode starch biosynthesis is one of the most important traits in lotus rhizome because of its relation to crop productivity. Understanding the microRNA (miRNA) and mRNA expression profiles related to lotus internode starch biosynthesis would help develop molecular improvement strategies, but they are not yet well-investigated. To identify genes and miRNAs involved in internode starch biosynthesis, the cDNA and small RNA libraries of Z6-1, Z6-2, and Z6-3 were sequenced, and their expression were further studied. Through combined analyses of transcriptome data and small RNA sequencing data, a complex co-expression regulatory network was constructed, in which 20 miRNAs could modulate starch biosynthesis in different internodes by tuning the expression of 10 target genes. QRT-PCR analysis, transient co-expression experiment and dual luciferase assay comprehensively confirmed that NnumiR396a down-regulated the expression of NnSS2 and ultimately prevents the synthesis of amylopectin, and NnumiR396b down-regulated the expression of NnPGM2 and ultimately prevents the synthesis of total starch. Our results suggest that miRNAs play a critical role in starch biosynthesis in lotus rhizome, and that miRNA-mediated networks could modulate starch biosynthesis in this tissue. These results have provided important insights into the molecular mechanism of starch biosynthesis in developing lotus rhizome.

BACKGROUND: The diagnosis of benign and malignant menopausal endometrial lesions (MEL) is often misled by complicated clinical indicators and ultrasonographic parameters in actual clinical applications. OBJECTIVE: To investigate the performance of clinical indicators and ultrasonographic parameters in the diagnosis of MEL. METHODS: A cohort of 156 enrolled menopausal patients with MEL was divided into benign group (128 cases) and malignant group (28 cases). Two clinical indicators of patient age (PA), abnormal vaginal bleeding (AVB) and three transvaginal ultrasonography (TVS) parameters of endometrial thickness (ET), endometrial uneven echo (EUE) and endometrial blood flow signal (EBFS) were measured for the mathematical modelling. The performance of combined indicators and individual indicators were firstly compared, and then the optimized combined indicators was compared with corresponding individual indicators, respectively. RESULTS: Our experiments verified that the mathematical modelling presented robust capabilities in the diagnosis of MEL with the sensitivity, specificity and AUC of 78.6%, 75.8% and 0.83 for combined indicators, and 75.0%, 81.3% and 0.85 for optimized combined indicators, respectively. The cut off thresholds of PA was 57.5 years, ET was 11.5 mm. Furthermore, the AVB presented the most important risk factor among the optimized indicators of PA, ET and AVB (P< 0.05). CONCLUSIONS: The combined indicators presented better performance in differentiating benign and malignant MEL and the AVB demonstrated the most capability for clinical applications.

This article investigated the role and the specific mechanism of Ruscogenin in Sjögren's syndrome (SS). NOD/ShiLtJ mice were treated with Ruscogenin, and acinar cells isolated from submandibular glands were treated with TNF-α, Ruscogenin and transfected with NLRP3 overexpression plasmid. Salivary flow rate (SFR) was measured at weeks 11, 13, 15, 17, and 20. Histological analysis of the submandibular glands was conducted by hematoxylin-eosin staining assay. IL-6, IL-17, TNF-α, and IL-1β mRNA expression was detected through qRT-PCR. AQP 5, AQP 4, P2X7R, NLRP3, caspase 1, IL-1β, Bax, and Bcl-2 protein levels were tested by western blot. Cell apoptosis was assessed through acridine orange and propidium iodide (AO/PI) staining assay and flow cytometry assay. Ruscogenin ameliorated the SFR and submandibular gland inflammation of NOD/ShiLtJ mice. Ruscogenin promoted the preservation of acinar cells and suppressed inflammation-related factors (P2X7R, NLRP3, caspase 1, and IL-1β) in submandibular gland tissues of NOD/ShiLtJ mice. Ruscogenin inhibited acinar cell apoptosis in NOD/ShiLtJ mice and reversed TNF-α-induced apoptosis and inflammation of acinar cells. NLRP3 overexpression reversed the repressive effect of Ruscogenin on TNF-α-induced inflammation and apoptosis of acinar cells. Ruscogenin ameliorated SS by inhibiting NLRP3 inflammasome activation.

To study the influence of total glucosides of paeony (TGP) on the expression of peripheral blood programmed cell death protein 1 (PD-1) and its ligand (PD-L1) in patients with primary Sjögren's syndrome (pSS).Ten patients with new-onset pSS were selected as the experimental group and were treated with 1.8 g of TGP (the main ingredient is Radix Paeoniae Alba) daily for 3 months; furthermore, 10 physically healthy individuals were selected as the control group. Peripheral blood mononuclear cells were isolated, and flow cytometry was used to detect PD-1 expression on the surface of CD4+ T and CD8+ T lymphocytes and PD-L1 expression on the surface of CD14+ monocytes and CD19+ B cells before and after treatment in the experimental and control groups. Furthermore, plasma levels of soluble PD-1 (sPD-1), interleukin (IL)-10, and IL-17A were also determined using enzyme-linked immunosorbent assay.The PD-1 expression on the surface of CD4+ T and CD8+ T lymphocytes in the peripheral blood of patients with pSS were significantly higher than in the control group (P0.001). However, PD-L1 expression on the surface of CD14+ monocytes declined but not significantly (P0.05), and PD-L1 expression on the surface of CD19+ B cells increased significantly (P0.001). Moreover, sPD-1 and IL-17A levels in the plasma of the experimental group were significantly higher than in the control group (P0.001), but the IL-10 level was significantly lower than in the control group (P0.001). After TGP treatment, PD-1 expression on the surface of CD4+ T and CD8+ lymphocytes in the peripheral blood of patients with pSS had decreased significantly (P0.001); the PD-L1 expression on the surface of CD19+ cells had decreased significantly (P0.001); and the PD-L1 expression on the surface of CD14+ monocytes did not differ significantly (P0.05). Furthermore, the levels of sPD-1 and IL-17A in plasma had decreased (P0.01) and IL-10 levels had increased after TGP treatment (P0.01).PD-1/PD-L1 molecules expressed on the surface of T cells, B cells, and monokaryon participated in the pathogenesis and development of SS through interactions. Therefore, TGP, which may increase the expression of PD-1 and its relevant ligand PD-L1 in the peripheral blood mononuclear cells, may play a role in the pathogenesis and development of SS through the PD-1/PD-L1 pathway by regulating regulatory T cells/T helper cell 17.

The aim of this study is to investigate the association between body composition, measured by bioelectrical impedance analysis, and outcomes in patients with acute kidney injury (AKI) receiving kidney replacement therapy (KRT).Patients with severe AKI treated with KRT in our hospital between September 2016 and August 2018 were enrolled. These patients were assessed by body composition analysis before KRT, and on the 3rd day and the 7th day after initiation of KRT. The predictors included lean tissue index (LTI), fat tissue index, and body cell mass index (BCMI). The association between all-cause mortality and predictors was analyzed using Cox regression.A total of 152 patients were included in this study, with a 28-day mortality of 46.7% and 1-year mortality of 60.5%. LTI (adjusted hazard ratio per standard deviation: 0.37; 95% confidence interval = 0.21-0.66, P .001) and BCMI (adjusted hazard ratio per standard deviation: 0.37; 95% confidence interval = 0.21-0.67, P .001) on day 7 after initiation of KRT, rather than before KRT, were associated with mortality during follow-up. LTI and BCMI before KRT were associated with 28-day mortality rather than 1-year mortality.LTI and BCMI before KRT were associated with short-term prognosis, and those on day 7 after KRT initiation were associated with intermediate mortality in patients with AKI requiring KRT.

Vitamin D deficiency is not only related to bone-related diseases but also associated with cardiovascular disease, diabetes, cancer and autoimmune diseases. Previous studies show that vitamin D deficiency increases risk of diabetic peripheral neuropathy (DPN). Data is limited among Chinese patients with type 2 diabetes. In this cross-sectional study, we enrolled 5362 patients (2359 men and 3003 women aged 30 to 89 years) with type 2 diabetes from Jan 1, 2016 to Oct 31, 2018. Serum 25-hydroxyvitamin D (25(OH)D) level was measured by electrochemiluminescence assay. Vitamin D deficiency was defined as Serum 25(OH)D level < 20 ng/mL according to the Endocrine Society Clinical Practice Guideline 2011. Diabetic peripheral neuropathy was diagnosed by motor/sensory nerve conduction velocity. Patients with diabetic peripheral neuropathy had lower serum 25(OH)D concentration (16.81±3.83 ng/mL vs. 18.05±4.11 ng/mL, P Disclosure L. Zhang: None. Y. Gu: None. Y. Yang: None. Y. Zhu: None. X. Yin: None. Y. Dong: None.

The current study aims to determine the secular trend of prevalence of type 2 diabetes and prediabetes and risk factors for diabetes from 2002 to 2009. We conducted 3 population-based surveys for diabetes in individuals aged 35-74 years in Qingdao in 2002, 2006 and 2009, respectively, with random-sampling approach. The sample size was 1792, 4549 and 4310 in 2002, 2006 and 2009, respectively. We performed a questionnaire-based survey to collect general information. All participants underwent standardized 2-h 75-g oral glucose tolerance tests (OGTTs). The 1999 WHO diagnostic criteria for diabetes and prediabetes was applied. Age-standardized prevalence of diabetes was calculated. Multiple non-conditional logistic regression were applied to analyze the risk factors for diabetes, taking diabetes as the dependent variable. In 2002, the age-standardized prevalence of diabetes was 12.32%, the corresponding prevalence was 16.57% and 18.01%, respectively, in 2006 and 2009. The prevalence of diabetes increased from 2002 to 2009 (p Disclosure L. Zhang: None. W. Gao: None. N. Wang: None. J. Wang: None. X. Yin: None. Y. Zhu: None. Y. Dong: None.

Aims: The aims of this study were to investigate the efficacy and identify the prognostic factors of continuous hemofiltration in patients with cardiorenal syndrome (CRS) and, finally, to optimally select patients who could benefit more from this therapy. Methods: A total of 59 patients with CRS type 1 or type 2 treated with continuous hemofiltration were enrolled. We collected their clinical data and divided them into 2 groups according to their survival or death during hospitalization to conduct a retrospective analysis on factors affecting mortality. Results: The following items were significantly different between the survival (n = 30) and death (n = 29) groups: serum creatinine, serum total bilirubin, direct bilirubin, white blood cells, hemoglobin, hematocrit, left ventricular end-diastolic diameter, left ventricular end-systolic diameter, systolic blood pressure and mean arterial pressure before hemofiltration, and average dehydration volume during the hemofiltration process. Leukocytosis was a risk factor for death (OR 1.242, 95% CI 1.242-1.480), and elevated sCr was not a key negative factor in the prognosis of CRS (OR 0.994, 95% CI 0.989-1.000). Conclusions: Cardiac function before hemofiltration and the amount of dehydration during the hemofiltration process both affected the prognosis. Infection and fluid overload condition at the beginning of hemofiltration were independently associated with mortality during hospitalization.

Background/Aims: To evaluate the long-term outcomes of Chinese patients with cardiac surgery-associated acute kidney injury (CSA-AKI). Methods: Patients who underwent cardiac surgery with a median 3-year follow-up were enrolled. The long-term survival rate and the incidence of chronic kidney disease (CKD) were recorded, and related risk factors were analyzed. Results: Of all 1,363 patients, 457 (33.5%) developed CSA-AKI. The AKI patients had a lower 3-year survival rate (88.8 vs. 97.2%, respectively, p < 0.001) and a higher incidence of CKD stages 3-5 (9.9 vs. 2.3%, respectively, p < 0.001) than the non-AKI patients. Cox regression analysis showed that AKI, atrial fibrillation, chronic cardiac insufficiency, longer surgical duration, respiratory failure after surgery, and longer mechanical ventilation time were associated with long-term mortality, while AKI, older age, and lower baseline kidney function were associated with incident CKD stages 3-5. Conclusion: CSA-AKI increased the risk of 3-year mortality and incident CKD stages 3-5.

Background/Aims: Renal replacement therapy (RRT) is a rescue therapy for patients with type 1 cardiorenal syndrome (CRS) with poor prognoses. However, the optimal timing for initiation and cessation of RRT remains controversial. The purpose of this study was to determine the optimal timing of initiation and cessation of RRT for patients with type 1 CRS. Methods: In this retrospective analysis, patients with refractory type 1 CRS receiving RRT were divided into 3 groups according to weaning from RRT and death within 90 days. Baseline characteristics, underlying heart disease, comorbidities, drug use before RRT, indicators of RRT initiation, and prognosis were compared between the 3 groups. Results: Fifty-two patients were enrolled, which included 27 males and 25 females with a mean age of 70.7 ± 16.1 years and a 90-day mortality rate of 65.4%. The mean urine output before RRT initiation was 800 mL/ 24 h in the RRT-independent group, 650 mL/24 h in the RRT-dependent group, and 345 mL/ 24 h in the death group (p = 0.021). Additionally, there were obvious differences in fluid balance between the 3 groups (167, 250, and 1,270 mL, respectively, p = 0.016). Patients could be successfully weaned from RRT when urine output was >880 mL and fluid balance volume was Conclusion: The mean fluid balance of survivors was remarkably less than that of the death group at RRT initiation. RRT termination can be considered when urine output is >880 mL/24 h and volume balance is

Background/Aims: The study aimed to investigate the relationship among mortality of patients with cardiac surgery-associated acute kidney injury (CSA-AKI), fluid balance, and ultrafiltration of renal replacement therapy (RRT). Methods: From January 2009 to October 2015, hospitalized patients with CSA-AKI receiving continuous or prolonged intermittent RRT were screened. The effects of fluid balance and ultrafiltration of RRT on clinical outcome were analyzed. Results: The 30-day mortality of all the 63 patients in the study was 58.6%. Compared with the death group, the survival group had a significantly lower fluid balance, larger ultrafiltration volume, and similar ultrafiltration rate during the first 3 days of RRT. Multivariate Cox regression analysis revealed that positive fluid balance during the first day of RRT, cardiac function of grade IV, and higher Sequential Organ Failure Assessment score were independent risk factors of 30-day mortality. Conclusion: Fluid balance was more relevant to short-term prognosis of CSA-AKI-RRT patients than ultrafiltration volume or ultrafiltration rate.

The key imaging parameter of number of diffusion gradient directions (NDGDs) in the protocol of diffusion tensor imaging (DTI) is significant in the acquisition of raw data. Up to date, the selection of NDGD in DTI is still in controversy for the optimized NDGD to observe 3D deterministic and probabilistic fiber trackings (FTs) of DTI in clinical evaluations. In this study, we evaluate the NDGDs of DTI on 3D deterministic and probabilistic fiber trackings (FTs). One volunteer was scanned by DTI protocols with five NDGDs of 6, 11, 15, 21, and 31 orientations, respectively. In 3D, the two deterministic and probabilistic FTs were qualitatively observed by delineating the region of interests (ROIs) of single-voxel and multiple-voxels. The results showed that the major fiber branches could be discovered with NDGD of six directions and the tracking performance could be easily affected by image noise; moreover, NDGD above 11 orientations could provide detailed fiber tract configurations for showing minor fiber tracts with longer scanning time.

Based on the absorption property of a diazine that can be formed by reaction of glyoxal and 3-methyl-2-benzothiazolinone hydrazone (MBTH) in the Ultraviolet-visible (UV-vis) spectral region, a HPLC method was developed for the determination of glyoxal in acetaldehyde solution. Glyoxal was derivatised from MBTH and the derivatives (diazine) were analyzed by HPLC for identification and quantification. The determination was performed on a ZORBAX Eclipse XDB-C18 column (4.6 × 250 mm, 5 μm) at 35°C with an injection volume of 10 μL, using a mixture of acetonitrile-water solvent (99:5, v:v) as a mobile phase with a flow rate of 0.8 mL·min−1. The proper derivative reaction conditions were the temperature of 70°C, MBTH to carbonyl molar ratio of 12, and reaction time of 110 min. The glyoxal diazine was a yellow dye with a maximum molar absorptivity at 401 nm and its retention time was 5.2 min under optimal HPLC conditions. The standard curve for glyoxal had a strong linear relationship with a regression coefficient (R2 = 0.999) in the range of 0.002–0.020 g·L−1. The analysis of glyoxal in an oxidising solution gave accurate results with a relative standard deviation (RSD) value of 0.55%. The average relative recovery was 102%. This efficient HPLC technique is also proposed for detecting other dicarbonyl compounds besides glyoxal.

The hydrazones have the absorption in the ultraviolet-visible (UV-vis) spectral region. The hydrazones can be formed by the reaction of aldehyde group and dinitrophenylhydrazine (DNPH) in acidic solution. The determination of glyoxal and glyoxalic acid in aldehyde solution was performed by high performance liquid chromatography (HPLC). The proper derivatization conditions were as follows: the reaction temperature of 70 degrees C, the pH of the system of 1.75; the molar ratio of DNPH to carbonyl of 6, the reaction time of 150 min. The saturated concentration of glyoxal dihydrazone in acetonitrile solution at pH 1.75 and 20 degrees C was 20.2 mg/L. The standard curves for glyoxal and glyoxalic acid both had good linear relations in the ranges of 2 - 20 mg/L and 10 - 100 mg/L, respectively. The glyoxal and glyoxalic acid contents in the solution of oxidation of acetaldehyde to glyoxal by nitric acid were determined by this method, the repeatabilities of analysis results were excellent and the relative error compared to chemical analysis of glyoxal was 1.77%. The spiked recoveries of glyoxal and glyoxalic acid were 99.6% - 103.3% and 98.1% - 102.4%, respectively. The method is simple, accurate and efficient for determining aldehydes and dicarbonyl compounds.

Objective The purpose of our study was to investigate the effect of coupled plasma filtration adsorption (CPFA) on endothelial cell (EC) function in patients with multiple organ dysfunction syndrome (MODS). Methods Besides routine therapy, the 24 MODS patients underwent both CPFA and high volume hemofiltration (HVHF), scheduled randomly at intervals of 12 hours. Patient serum from 0, 5, and 10 hours of therapy was collected to measure soluble E-selectin (sE-selectin) and soluble thrombomodulin (sTM) by the ELISA method. Human umbilical vein endothelial cells (HUVEC) were incubated for 24 hours with the patient serum and the supernatant liquid was gathered to detect sTM and sE-selectin. The proliferation function of the ECs was detected by methyl thiazolyl tetrazolium (MTT) method. Results 1. The serum levels of sE-selectin and sTM were significantly higher in MODS patients than in controls; serum sE-selectin and sTM decreased remarkably after a single circulation in CPFA (p0. 05); the level of sE-selectin and sTM in systemic circulation had no change during CPFA or HVHF (p>0.05); 2. sTM in supernatant liquid incubated with serum from 5 hours of CPFA and 10 hours of HVHF decreased remarkably (p0. 05); 3. when incubated with serum taken from the device pre- or post-CPFA, the optical density (OD) value of the latter was higher. The OD value increased gradually when incubated with serum from 0, 5, and 10 hours of CPFA (pConclusions CPFA can eliminate sE-selectin and sTM and improve the secretion function of ECs. CPFA was somewhat better and earlier than HVHF, while to a certain degree it can weaken the inhibitory effect of serum on the proliferation function of ECs.

The hydrazones have the absorption in the ultraviolet-visible (UV-vis) spectral region. The hydrazones can be formed by the reaction of aldehyde group and dinitrophenylhydrazine (DNPH) in acidic solution. The determination of glyoxal and glyoxalic acid in aldehyde solution was performed by high performance liquid chromatography (HPLC). The proper derivatization conditions were as follows: the reaction temperature of 70 degrees C, the pH of the system of 1.75; the molar ratio of DNPH to carbonyl of 6, the reaction time of 150 min. The saturated concentration of glyoxal dihydrazone in acetonitrile solution at pH 1.75 and 20 degrees C was 20.2 mg/L. The standard curves for glyoxal and glyoxalic acid both had good linear relations in the ranges of 2 - 20 mg/L and 10 - 100 mg/L, respectively. The glyoxal and glyoxalic acid contents in the solution of oxidation of acetaldehyde to glyoxal by nitric acid were determined by this method, the repeatabilities of analysis results were excellent and the relative error compared to chemical analysis of glyoxal was 1.77%. The spiked recoveries of glyoxal and glyoxalic acid were 99.6% - 103.3% and 98.1% - 102.4%, respectively. The method is simple, accurate and efficient for determining aldehydes and dicarbonyl compounds.