Objective To investigate the effects of nephroblastoma over-expressed protein (CCN3) on the formation of extracellular matrix (ECM) induced by transforming growth factor-β1 (TGF-β1) in human mesangial cells (HMCs) and its underlying signal transduction mechanism related with microRNA-29(miRNA-29).Methods HMCs were pretreated with different doses of exogenous CCN3 (5 μg/L,50 μg/L and 500 μg/L) or transfected with pcDNA3.1(+)-CCN3 before exposed to TGF-β1(2 μg/L),to observe the expression of fibronectin (FN),type I collagen (COL I) and miRNA-29a,b and c.The mimics or inhibitor of the miRNA-29a were transfected into HMCs to analyze whether miRNA-29a affect CCN3.The expressions of FN mRNA,COL I mRNA and miRNA-29 family were detected by real time PCR.The protein expressions of FN and COL I were detected by Western blotting and cell immunofluorescence.Results (1) Compared with the normal control group,the expressions of FN and COL I were up-regulated in TGF-β1 group,while the expressions of miRNA-29a,b,c were down-regulated in TGF-β1 group (all P ＜ 0.05).(2) Compared with the TGF-β1 group,the expressions of FN and COL I were decreased when pretreated with the different doses of exogenous of CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P ＜ 0.05).Meanwhile,the expression of miRNA-29a was significantly increased when pretreated with 50 μg/L and 500 μg/L CCN3 or transfected with pcDNA3.1(+)-CCN3 (all P ＜ 0.05);whereas miRNA-29b and c had no statistical difference (all P ＞ 0.05).(3) Compared with TGF-β1+CCN3 group,the expressions of FN and COL I were decreased in CCN3+TGF-β1+miRNA-29a mimics group (all P ＜ 0.05),whereas the expressions of FN and COL I in CCN3+TGF-β1+miRNA-29a inhibitors group were increased (all P ＜ 0.05).Conclusions CCN3 reduces the TGF-β1-induced production of ECM by the up-regulation of miRNA-29a.

Objective To investigate the curative effect of lateral position ventilation combined with vibration sputum drainage on the patients with acute respiratory distress syndrome (ARDS). Methods A prospective randomized controlled trial was conducted. The patients with ARDS undergoing mechanical ventilation (MV) admitted to intensive care unit (ICU) of Linyi Central Hospital from January 2013 to June 2017 were enrolled, and they were divided into simple ventilation group and combined treatment group according to random number table. The patients in both groups received etiological treatment, protective ventilation strategy, sensitive antibiotics for anti-infection, and calefacient and humidifying treatment. The patients in the simple ventilation group received bilateral discontinuous alternative lateral position ventilation [pressure controlled ventilation (PCV), tidal volume (VT) ≤ 6 mL/kg, the inhaled oxygen concentration (FiO2) and positive end expiratory pressure (PEEP) was adjusted to maintain the airway plateau pressure (Pplat) ≤ 30 cmH2O (1 cmH2O = 0.098 kPa)], and those in the combined treatment group received lateral position ventilation combined with vibration sputum drainage, twice a day, 15 minutes each time. The parameters of respiratory function and inflammation as well as excretion of sputum before and after treatment were compared between the two groups, and the complication and prognosis were recorded. Results A total of 200 patients with ARDS were included, 4 patients were excluded because of severe pneumothorax, massive hemorrhage of the digestive tract, or elevated intracranial pressure, so 196 patients were finally enrolled in the analysis. There were 98 patients in the simple ventilation group and the combined treatment group, respectively. There were no significant differences in parameters of respiratory function and inflammation as well as excretion of sputum before treatment between the two groups. After treatment, the above parameters in both groups were improved, and the effect of combined treatment was more significant. Compared with the simple ventilation group, white blood cell count (WBC), procalcitonin (PCT), C-reactive protein (CRP) at 24 hours of treatment in the combined treatment group were significantly decreased [WBC (×109/L):9.1±1.6 vs. 11.8±3.6, PCT (μg/L): 14.5±2.4 vs. 22.7±3.2, CRP (mg/L): 32.2±6.3 vs. 67.2±7.2, all P < 0.01], the quantity of sputum excretion was significantly increased (mL: 49.3±12.5 vs. 36.8±11.8, P < 0.01); 72 hours after treatment, the oxygenation index (PaO2/FiO2) in the combined treatment group was significantly increased [mmHg (1 mmHg = 0.133 kPa): 278±28 vs. 238±39, P < 0.01], and PEEP, FiO2, airway resistance (Raw) were significantly lowered [PEEP (cmH2O): 5±2 vs. 7±3, FiO2: 0.40±0.11 vs. 0.49±0.12, Raw (cmH2O): 12.8±0.7 vs. 16.2±0.8, all P < 0.01]. There was no significant difference in static lung compliance (Cst) or Pplat between the two groups. Compared with the simple ventilation group, the duration of MV (days: 5.5±3.0 vs. 8.6±2.1), the length of ICU stay (days: 7.5±5.4 vs. 11.3±4.6) and the extraction time of artificial airway (days: 6.6±2.8 vs. 9.4±3.3) in the combined treatment group were significantly shortened (all P < 0.01). However, there was no significant difference in the 28-day mortality rate [4.1% (4/98) vs. 3.1% (3/98)], the rate of tracheotomy [2.0% (2/98) vs. 3.1% (3/98)] or the incidence ofⅠ-Ⅱ skin pressure sore [1.0% (1/98) vs. 2.0% (2/98)] between simple ventilation group and the combined treatment group (all P > 0.05). No artificial airway disappearance, pneumothorax, grade Ⅲ or Ⅳ pressure sore was found in the two groups. Conclusions Compared with lateral position ventilation alone, the treatment of combined with vibration sputum drainage is more effective for improving the respiratory function of patients with ARDS, reducing infection action, shortening the duration of MV and the length of ICU stay, and improve the recovery, promote the rehabitation of patients.

Objective:To investigate the risk factors of thrombocytopenia in elderly patients with sepsis-associated acute kidney injury(SA-AKI), and to further evaluate whether the degree of thrombocytopenia is related to the increased risk of death on day 28.Methods:Elderly patients with SA-AKI admitted to ICU of our hospital from June 2017 to June 2020 were selected.The patients were divided into normal platelet group(56 cases)and thrombocytopenia group(72 cases)according to the platelet(PLT)count, and according to the degree of thrombocytopenia, they were further divided into three groups: PLT<20×10 9/L(group A, 22 cases), 20×10 9/L≤ PLT<50×10 9/L(group B, 27 cases), 50×10 9/L≤ PLT<100×10 9/L(group C, 23 cases).The general data, clinical baseline indicators and prognostic indicators of each group were compared to evaluate the risk factors of thrombocytopenia.At the same time, the impact of platelet count on the prognosis of elderly patients with SA-AKI was evaluated according to the length of stay in ICU, total length of stay and whether the patient died, and the correlation between the degree of thrombocytopenia and survival was analyzed. Results:A total of 128 elderly patients with SA-AKI were enrolled, including 73 males and 55 females.59.4 % of the patients were hospitalized in the department of internal medicine.The APACHEⅡ score was(15.5 ± 3.3)points, invasive mechanical ventilation accounted for 78.9%, positive inotropic therapy accounted for 12.8%, and 56 patients had normal platelet count.Thrombocytopenia occurred in 72 patients, including 22 patients with PLT<20×10 9/L, 27 patients with platelet count in 20-50×10 9/L, and 23 patients with platelet count in 50-100×10 9/L.There were significant differences in bloodstream infection, Gram-negative bacteria, APACHEⅡ score and procalcitonin(PCT)among the four groups( P<0.05).Further multivariate logistic regression showed that PCT was an independent risk factor for thrombocytopenia in elderly patients with SA-AKI( OR=1.05, 95% CI: 1.00-1.10, P=0.042).Compared with the normal platelet group, the 28-day mortality rate of the thrombocytopenia group was significantly higher than that of the normal platelet group, while the length of stay in ICU and the total length of stay were prolonged( P<0.05).Kaplan-Meier survival curve showed that the 28-day cumulative survival rate of thrombocytopenia group was lower than that of normal platelet group, and the risk of death increased with the degree of thrombocytopenia( χ2=31.479, P<0.001).Univariate Cox regression analysis showed that thrombocytopenia, bloodstream infection, APACHEⅡ score and PCT were risk factors for 28-day death in elderly patients with SA-AKI.Multivariate Cox analysis showed that bloodstream infection and APACHEⅡ score were independent risk factors for 28-day death.After adjusting these confounding factors, thrombocytopenia was an independent risk factor for death, and the degree of thrombocytopenia was related to the increased risk of death. Conclusions:Thrombocytopenia is common in elderly patients with SA-AKI, and elevated PCT levels increase the risk of thrombocytopenia.The degree of thrombocytopenia is an independent risk factor for 28-day mortality in such patients.

Reversible phosphorylation plays a crucial role in regulating protein activities and functions. Sexual reproduction directly affects yield of most agricultural crops. As the male reproductive organ, anther generates microspores (pollen), delivering gametes (sperms) to complete double fertilization in higher plants. Here, we took the advantage of Nano UHPLC-MS/MS to analyze maize (Zea mays, B73) early anthers at proteomic and phosphoproteomic levels, to explore the protein and phosphorylation modification regulatory networks controlling maize anther development. Our proteomic analysis identified 3 016 unique peptides, belonging to 1 032 maize proteins. MapMan analysis revealed variously potential proteins associated with maize anther development, such as receptor-like kinases (GRMZM2G082823_P01 and GRMZM5G805485_P01). Using phospho-peptides enriched by TiO2 affinity chromatography, our phosphoproteomic analysis detected 257 phospho-peptides from 210 phosphoproteins, discovering 223 phosphosites. Compared to the 86 maize phosphoproteins collected in the Plant Protein Phosphorylation Data Base (P3DB), we found that 203 phosphoproteins and 218 phosphosites were not revealed before. Further bioinformatics analysis revealed that phosphorylation of 14-3-3 proteins, kinases, phosphatases, transcription factors, cell cycle and chromatin structure related proteins might play important roles in regulating normal anther development in maize. Our findings not only enlarged the maize phosphoproteome data, but also provided information for analyzing the molecular mechanism controlling maize anther development at genetic and biochemical levels.
Crops, Agricultural ; chemistry ; Phosphoproteins ; chemistry ; Phosphorylation ; Plant Proteins ; chemistry ; Pollen ; chemistry ; Proteome ; Tandem Mass Spectrometry ; Zea mays ; chemistry