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Plant and fruit surfaces are covered with cuticle wax and provide a protective barrier against biotic and abiotic stresses. Cuticle wax consists of very-long-chain fatty acids (VLCFAs) and their derivatives. ß-Ketoacyl-CoA synthase (KCS) is a key enzyme in the synthesis of VLCFAs and provides a precursor for the synthesis of cuticle wax, but the KCS gene family was yet to be reported in the passion fruit (Passiflora edulis). In this study, thirty-two KCS genes were identified in the passion fruit genome and phylogenetically grouped as KCS1-like, FAE1-like, FDH-like, and CER6-like. Furthermore, thirty-one PeKCS genes were positioned on seven chromosomes, while one PeKCS was localized to the unassembled genomic scaffold. The cis-element analysis provides insight into the possible role of PeKCS genes in phytohormones and stress responses. Syntenic analysis revealed that gene duplication played a crucial role in the expansion of the PeKCS gene family and underwent a strong purifying selection. All PeKCS proteins shared similar 3D structures, and a protein-protein interaction network was predicted with known Arabidopsis proteins. There were twenty putative ped-miRNAs which were also predicted that belong to nine families targeting thirteen PeKCS genes. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation results were highly associated with fatty acid synthase and elongase activity, lipid metabolism, stress responses, and plant-pathogen interaction. The highly enriched transcription factors (TFs) including ERF, MYB, Dof, C2H2, TCP, LBD, NAC, and bHLH were predicted in PeKCS genes. qRT-PCR expression analysis revealed that most PeKCS genes were highly upregulated in leaves including PeKCS2, PeKCS4, PeKCS8, PeKCS13, and PeKCS9 but not in stem and roots tissues under drought stress conditions compared with controls. Notably, most PeKCS genes were upregulated at 9th dpi under Fusarium kyushuense biotic stress condition compared to controls. This study provides a basis for further understanding the functions of KCS genes, improving wax and VLCFA biosynthesis, and improvement of passion fruit resistance.
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Objective:To investigate the correlation between the expressions of miR-524-5p and sex determining region Y box protein 9 (SOX9) in advanced gastric cancer and their influences on the efficacy and prognosis of chemotherapy.Methods:A total of 82 patients diagnosed as advanced gastric cancer who received DCF (docetaxel + cisplatin + fluorouracil) chemotherapy in 910th Hospital of Joint Logistics Support Force of Chinese People′s Liberation Army from January 2015 to December 2017 were selected as the research objects. The expression levels of miR-524-5p and SOX9 in gastric cancer tissues were detected by fluorescence quantitative PCR. The correlation between the expressions of miR-524-5p and SOX9 was analyzed, the total effective rates of chemotherapy in patients with different miR-524-5p and SOX9 expression levels were compared, and the predictive value of miR-524-5p combined with SOX9 on the efficacy of chemotherapy was analyzed. Overall survival (OS) and progression-free survival (PFS) were analyzed.Results:The expressions of miR-524-5p and SOX9 were not related to gender or age of patients with advanced gastric cancer (all P>0.05), but were related to the degree of differentiation ( χ2=3.577, P=0.001; χ2=5.654, P<0.001) and distant metastasis ( χ2=2.466, P=0.016; χ2=5.218, P<0.001) of patients with advanced gastric cancer. There was a negative correlation between the expressions of miR-524-5p and SOX9 in advanced gastric cancer ( r=-0.348, P=0.001). According to the median expressions of miR-524-5p and SOX9 in gastric cancer tissues, patients were divided into high expression and low expression groups, miR-524-5p≥0.64 was high expression ( n=41), <0.64 was low expression ( n=41), SOX9≥1.84 was high expression ( n=41), and <1.84 was low expression ( n=41). The total effective rate of advanced gastric cancer patients with high expression of miR-524-5p was 58.54% (24/41), which was higher than that of patients with low expression of miR-524-5p (24.39%, 10/41), and there was a statistically significant difference ( χ2=9.484, P=0.002). The total effective rate of advanced gastric cancer patients with high expression of SOX9 was 21.95% (9/41), which was lower than 60.97% (25/41) of patients with low expression of SOX9, and there was a statistically significant difference ( χ2=12.863, P<0.001). Receiver operating characteristic curve analysis showed that the expressions of miR-524-5p and SOX9 had predictive value for the efficacy of chemotherapy, and the area under the curve was 0.753 (95% CI: 0.644-0.861, P<0.001) and 0.660 (95% CI: 0.540-0.780, P=0.014) respectively. The combination of miR-524-5p and SOX9 had predictive value for the efficacy of chemotherapy, and the area under the curve was 0.768 (95% CI: 0.667-0.868, P<0.001). The median PFS and OS of patients with high expression of miR-524-5p were 8 months and 14 months, which were longer than those of patients with low expression of miR-524-5p (6 months, 9 months), and there were statistically significant differences ( χ2=21.160, P<0.001; χ2=29.730, P<0.001). The median PFS and OS of patients with high expression of SOX9 were 7 months and 10 months, which were shorter than those of patients with low expression of SOX9 (8 months, 12 months), and there were statistically significant differences ( χ2=6.345, P=0.012; χ2=4.107, P=0.043). Conclusion:There is a negative correlation between the expressions of miR-524-5p and SOX9 in advanced gastric cancer tissues. The chemotherapy efficacy and prognosis of patients with high expression of miR-524-5p and low expression of SOX9 are better than those of patients with low expression of miR-524-5p and high expression of SOX9.
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Production of passion fruit (Passiflora edulis) is restricted by postharvest decay, which limits the storage period. We isolated, identified, and characterized fungal pathogens causing decay in two passion fruit cultivars during two fruit seasons in China. Morphological characteristics and nucleotide sequences of ITS-rDNA regions identified eighteen isolates, which were pathogenic on yellow and purple fruit. Fusarium kyushuense, Fusarium concentricum, Colletotrichum truncatum, and Alternaria alternata were the most aggressive species. Visible inspections and comparative analysis of the disease incidences demonstrated that wounded and non-wounded yellow fruit were more susceptible to the pathogens than the purple fruit. Purple cultivar showed higher expression levels of defense-related genes through expression and metabolic profiling, as well as significantly higher levels of their biosynthesis pathways. We also found fungi with potential beneficial features for the quality of fruits. Our transcriptomic and metabolomics data provide a basis to identify potential targets to improve the pathogen resistance of the susceptible yellow cultivar. The identified fungi and affected features of the fruit of both cultivars provide important information for the control of pathogens in passion fruit industry and postharvest storage.
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Objective:To compare the difference of low-level assisted ventilation and T-piece method on respiratory mechanics of patients with invasive mechanical ventilation during spontaneous breathing trial (SBT) within 3 days before extubation.Methods:A retrospective observational study was conducted. Twenty-five patients with difficulty in weaning or delayed weaning from invasive mechanical ventilation who were admitted to department of critical care medicine of the First Affiliated Hospital of Guangzhou Medical University from December 2018 to June 2020, and were in stable condition and entered the weaning stage after more than 72 hours of invasive mechanical ventilation were studied. A total of 119 cases of respiratory mechanical indexes were collected, which were divided into the low-level assisted ventilation group and the T-piece group according to the ventilator method and parameters used during the data collection. The different ventilation modes related respiratory mechanics indexes such as the esophageal pressure (Pes), the gastric pressure (Pga), the transdiaphragmatic pressure (Pdi), the maximum Pdi (Pdimax), Pdi/Pdimax ratio, the esophageal pressure-time product (PTPes), the gastric pressure-time product (PTPga), the transdiaphragmatic pressure-time product (PTPdi), the diaphragmatic electromyography (EMGdi), the maximum diaphragmatic electromyography (EMGdimax), PTPdi/PTPes ratio, Pes/Pdi ratio, the inspiratory time (Ti), the expiratory time (Te) and the total time respiratory cycle (Ttot) at the end of monitoring were recorded and compared between the two groups.Results:Compared with the T-piece group, Pes, PTPes, PTPdi/PTPes ratio, Pes/Pdi ratio and Te were higher in low-level assisted ventilation group [Pes (cmH 2O, 1 cmH 2O = 0.098 kPa): 2.84 (-1.80, 5.83) vs. -0.94 (-8.50, 2.06), PTPes (cmH 2O·s·min -1): 1.87 (-2.50, 5.93) vs. -0.95 (-9.71, 2.56), PTPdi/PTPes ratio: 0.07 (-1.74, 1.65) vs. -1.82 (-4.15, -1.25), Pes/Pdi ratio: 0.17 (-0.43, 0.64) vs. -0.47 (-0.65, -0.11), Te (s): 1.65 (1.36, 2.18) vs. 1.33 (1.05, 1.75), all P < 0.05], there were no significant differences in Pga, Pdi, Pdimax, Pdi/Pdimax ratio, PTPga, PTPdi, EMGdi, EMGdimax, Ti and Ttot between the T-piece group and the low-level assisted pressure ventilation group [Pga (cmH 2O): 6.96 (3.54,7.60) vs. 7.74 (4.37, 11.30), Pdi (cmH 2O): 9.24 (4.58, 17.31) vs. 6.18 (2.98, 11.96), Pdimax (cmH 2O): 47.20 (20.60, 52.30) vs. 29.95 (21.50, 47.20), Pdi/Pdimax ratio: 0.25 (0.01, 0.34) vs. 0.25 (0.12, 0.41), PTPga (cmH 2O·s·min -1): 7.20 (2.54, 9.97) vs. 7.97 (5.74, 13.07), PTPdi (cmH 2O·s·min -1): 12.15 (2.95, 19.86) vs. 6.87 (2.50, 12.63), EMGdi (μV): 0.05 (0.03, 0.07) vs. 0.04 (0.02, 0.06), EMGdimax (μV): 0.07 (0.05, 0.09) vs. 0.07 (0.04, 0.09), Ti (s): 1.20 (0.95, 1.33) vs. 1.07 (0.95, 1.33), Ttot (s): 2.59 (2.22, 3.09) vs. 2.77 (2.35, 3.24), all P > 0.05]. Conclusions:When mechanically ventilated patients undergo SBT, the use of T-piece method increases the work of breathing compared with low-level assisted ventilation method. Therefore, long-term use of T-piece should be avoided during SBT.
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BackgroundIn December 2019, human infection with a novel coronavirus, known as SARS-CoV-2, was identified in Wuhan, China. The mortality of critical illness was high in Wuhan. Information about critically ill patients with SARS-CoV-2 infection outside of Wuhan is scarce. We aimed to provide the clinical features, treatment, and prognosis of the critically ill patients with SARS-CoV-2 infection in Guangdong Province. MethodsIn this multi-centered, retrospective, observational study, we enrolled critically ill patients with SARS-CoV-2 pneumonia who were admitted to the intensive care unit (ICU) in Guangdong Province. Demographic data, symptoms, laboratory findings, comorbidities, treatments, and prognosis were collected. Data were compared between patients with and without intubation. ResultsForty-five critically ill patients with SARS-CoV-2 pneumonia were identified in 7 ICUs in Guangdong Province. The mean age was 56.7 years, and 29 patients (64.4%) were men. The most common symptoms at the onset of illness were high fever and cough. Majority of patients presented with lymphopenia and elevated lactate dehydrogenase. Treatment with antiviral drugs was initiated in all the patients. Thirty-seven patients (82.2%) had developed acute respiratory distress syndrome, and 13 (28.9%) septic shock. A total of 20 (44.4%) patients required intubation and 9 (20%) required extracorporeal membrane oxygenation. As of February 28th 2020, only one patient (2.2%) had died and half of them had discharged of ICU. ConclusionsInfection with SARS-CoV-2 in critical illness is characterized by fever, lymphopenia, acute respiratory failure and multiple organ dysfunction. Compared with critically ill patients infected with SARS-CoV-2 in Wuhan, the mortality of critically ill patients in Guangdong Province was relatively low. These data provide some general understandings and experience for the critical patients with SARS-CoV-2 outside of Wuhan.
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BackgroundAdministration of convalescent plasma may be of clinical benefit for treatment of severe acute viral respiratory infections. However, no clear evidence exists to support or oppose convalescent plasma use in clinical practice. We conducted a systematic review and meta-analysis to assess the evidence of randomized controlled trials (RCTs) in the convalescent plasma for the treatment of severe influenza. MethodsHealthcare databases were searched in February 2020. All records were screened against the eligibility criteria. Data extraction and risk of bias assessments were undertaken. The primary outcome was case-fatality rates by influenza. ResultsWe identified 5 RCTs of severe influenza. The pooled analyses showed no evidence for a reduction in mortality (Odds Ratio (OR) 1.06; 95% confidence interval (CI) 0.51-2.23; p = 0.87; I2 = 35%). We also found non-significant reductions in days in ICU and hospital, and days on mechanical ventilation. There seemed to have a biological benefit of increasing HAI titer levels and decreasing influenza B virus loads and cytokines after convalescent plasma treatment. No serious adverse events was reported between two groups. Studies were commonly of low risk of bias with high quality. ConclusionsConvalescent plasma appears safe but may not reduce mortality in severe influenza. This therapy should be studied within the context of a well-designed clinical trial for treatment of SARS-Cov-2 infection.
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Objective@#To examine the prevalence and associated factors of seeking casual male sexual partners among men who have sex with men (MSM) students in Guangzhou, and to provide a reference for health education and intervention.@*Methods@#A cross-sectional study was conducted to recruit 189 MSM students aged 18 and 28 via voluntary counseling and testing (VCT) clinics, Lingnan partners and Zhitong charity from May 2017 to April 2018. An anonymous electronic questionnaire was administrated to collect information on sociodemographic and sexual behavior characteristics. Chi-square test and non-conditional Logistic regression analysis were performed to explore the associated factors of casual homosexual behavior among MSM students.@*Results@#In the past 6 months, 63.49% (120/189) of them reported casual male partners, and the rate of consistent condom use was 61.67%(74/120). Multivariate Logistic regression analysis showed that participants who had multiple sex partners in the past 6 months were more likely to seek male casual partners(aOR=9.13, 95%CI=4.27-19.53). Individuals who used recreational drugs before sex were at a higher risk of having casual sex behaviors(aOR=3.06, 95%CI=1.08-8.62). MSM students who had steady homosexual partners had a lower risk to seek casual sex(aOR=0.43, 95%CI=0.19-0.98).@*Conclusion@#The proportion of seeking casual male sexual partners among MSM students in Guangzhou was relatively high. The publicity and education should be strengthened among those who had multiple sex partners, with no steady homosexual partners and used recreational drugs before sex, to reduce the casual sex behaviors.
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Objective To explore the effect and mechanism of Sacubitril/Valsartan on myocardial remodeling and cardiac function in rats with myocardial infarction.Methods The acute myocardial infarction (AMI) rat model was established by ligating anterior descending branch of coronary artery for one week.A total of 60 adult male rats in SPF grade with AMI were randomized into the Sacubitril/Valsartan group and the model group,who were gavaged with Sacubitril/Valsartan (68 mg/kg,once daily,n=30) versus with normal saline once daily(n=30) for 4 weeks.Twenty-four hours after the last treatment,the left ventricular cardiac function was examined by echocardiography,and pathological changes of the left ventricle were observed under light microscope.The degree of myocardial fibrosis was quantitatively analyzed by picric acid-sirius scarlet staining.Myocardial cells and fibroblasts from rat pups of the same species were prepared in vitro and were divided into the control group,Ang Ⅱ group,LBQ657 group,valsartan group and LCZ696 group.3 [H]-leucine incorporation and 3[H]-proline incorporation were used to detect the myocardial hypertrophy and fibrosis.Results There was no significant difference in left ventricular function between the the model group and the Sacubitril/Valsartan group before medication (P ＞ 0.05).Four weeks after administration of the medications,end-diastolic diameter of left ventricle and end-systolic volume of left ventricle were lower [(9.73±0.26) mm vs.(10.52±0.21) mm,P＜0.05;(0.19±0.03) ml vs.(0.31±0.02) ml,P＜0.01],and the left ventricular ejection fraction was higher [(60.17±2.18)％vs.(47.16± 5.14)％,P＜0.01] in the Sacubitril/Valsartan group than in the model group.The degree of myocardial cell injury in the infarct area was lower,and the area of myocardial fibrosis in the non-infarct zone and peripheral infarcted zone were less in the Sacubitril/Valsartan group than in the modelgroup[(4.0±0.1)％ vs.(6.1±0.8)％,P＜0.001;(15.7±0.8)％ vs.(23.8±1.2)％,P＜0.001].3 [H]-proline incorporation in cardiac fibroblasts was lower in the Valsartan group than in theAng Ⅱ group [(152.77±8.46) CPM vs.(221.87±13.41) CPM,P＜0.01].3[H]-leucine incorporation in myocardial cells was lower in the Valsartan group than in the Ang Ⅱ group [(113.47 ±2.33) CPM vs.(127.65 ± 2.38) CPM,P＜0.01].3 [H]-leucine incorporation in myocardial cells was lower in LBQ657 group than in the Ang Ⅱ group [(119.78±2.98) CPM vs.(127.65±2.38)CPM,P＜0.05],and the combined application of valsartan and LBQ657 can further reduce myocardial hypertrophy and fibrosis (P ＜ 0.05).Conclusions Sacubitril/Valsartan can effectively alleviate myocardial remodeling and cardiac dysfunction after myocardial infarction,and the mechanism may be related to reducing Ang Ⅱ-induced myocardial hypertrophy and myocardial fibrosis.
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Objective@#To explore the effect and mechanism of Sacubitril/Valsartan on myocardial remodeling and cardiac function in rats with myocardial infarction.@*Methods@#The acute myocardial infarction (AMI) rat model was established by ligating anterior descending branch of coronary artery for one week.A total of 60 adult male rats in SPF grade with AMI were randomized into the Sacubitril/Valsartan group and the model group, who were gavaged with Sacubitril/Valsartan (68 mg/kg, once daily, n=30) versus with normal saline once daily(n=30) for 4 weeks.Twenty-four hours after the last treatment, the left ventricular cardiac function was examined by echocardiography, and pathological changes of the left ventricle were observed under light microscope.The degree of myocardial fibrosis was quantitatively analyzed by picric acid-sirius scarlet staining.Myocardial cells and fibroblasts from rat pups of the same species were prepared in vitro and were divided into the control group, AngⅡ group, LBQ657 group, valsartan group and LCZ696 group.3[H]-leucine incorporation and 3[H]-proline incorporation were used to detect the myocardial hypertrophy and fibrosis.@*Results@#There was no significant difference in left ventricular function between the the model group and the Sacubitril/Valsartan group before medication (P>0.05). Four weeks after administration of the medications, end-diastolic diameter of left ventricle and end-systolic volume of left ventricle were lower [(9.73±0.26) mm vs.(10.52±0.21) mm, P<0.05; (0.19±0.03) ml vs.( 0.31±0.02) ml, P<0.01], and the left ventricular ejection fraction was higher [(60.17±2.18)% vs.(47.16±5.14)%, P<0.01] in the Sacubitril/Valsartan group than in the model group.The degree of myocardial cell injury in the infarct area was lower, and the area of myocardial fibrosis in the non-infarct zone and peripheral infarcted zone were less in the Sacubitril/Valsartan group than in the model group [(4.0±0.1)% vs. (6.1±0.8)%, P<0.001; (15.7±0.8)% vs. (23.8±1.2)%, P<0.001].3[H]-proline incorporation in cardiac fibroblasts was lower in the Valsartan group than in the Ang Ⅱ group [(152.77±8.46) CPM vs.(221.87±13.41) CPM, P<0.01].3[H]-leucine incorporation in myocardial cells was lower in the Valsartan group than in the Ang Ⅱ group [(113.47±2.33) CPM vs.(127.65±2.38) CPM, P<0.01].3[H]-leucine incorporation in myocardial cells was lower in LBQ657 group than in the Ang Ⅱ group [(119.78±2.98) CPM vs.(127.65±2.38) CPM, P<0.05], and the combined application of valsartan and LBQ657 can further reduce myocardial hypertrophy and fibrosis (P<0.05).@*Conclusions@#Sacubitril/Valsartan can effectively alleviate myocardial remodeling and cardiac dysfunction after myocardial infarction, and the mechanism may be related to reducing AngⅡ-induced myocardial hypertrophy and myocardial fibrosis.
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Objective To investigate the effects of Dingkoulizhong decoction on cellular immune factors and adverse reactions in patients with advanced gastric cancer treated with chemotherapy.Methods One hundred and thirty-eight patients with advanced gastric cancer were enrolled in our hospital from October 2014to December 2016,and were randomly divided into control group (n =69) and observation group (n =69) by using the random number table.The patients of the control group were treated with FOLFOX4 chemotherapy (oxaliplatin + calcium folinate + fluorouracil),while the patients of the observation group were given by the treatment of Dingkoulizhong decoction on the basis of the control group.The peripheral blood samples of the patients were collected before and after the treatment.The levels of cellular immune factors CD3 +,CD4 +,CD8 + and CD4 +/CD8 + were detected,and the clinical efficacy and adverse reactions of the patients of the two groups were observed.Results After the treatment,the number of complete remission (CR),partial remission (PR),stable disease (SD) and progressive disease (PD) in the observation group were 16,25,16 and 12 cases respectively,while the control group were 9,20,19 and 21 cases respectively,and there was a statistically significant difference in the distribution of clinical efficacy between the two groups (Z =4.381,P =0.036).Compared with the control group,the clinical benefit rate (CBR) of the observation group was significantly improved (59.42％ vs.42.03％),with a statistically significant difference (x2 =4.175,P =0.041).The cellular immune factors CD3 + [(52.67 ±6.21)％ vs.(53.45 ±6.54)％],CD4 + [(23.56 ±3.85)％ vs.(24.09±2.91)％],CD8 +[(28.16±3.49)％ vs.(27.87±3.26)％] and CD4 +/CD8+(1.13 ± 0.27 vs.1.19 ±± 0.31) of the patients of the observation group showed no statistically significant difference (t=0.718,P=0.474;t=0.912,P=0.363;t=0.504,P=0.615;t=1.212,P=0.227) beforeand after the treatment,but the cellular immune factor CD3 + [(50.36 ± 3.74)％ vs.(53.13 ± 6.12)％],CD4 +[(21.26±2.37)％ vs.(23.44 ±3.96)％] andCD4+/CD8*(0.96±0.26vs.1.15±0.25) of the patients of the control group after the treatment were significantly lower than those before the treatment,and CD8 +[(31.64 ± 4.05) ％ vs.(27.98 ± 3.52) ％] after the treatment was significantly higher than that before the treatment,all with statistically significant differences (t =3.208,P ＜ 0.001;t =3.924,P ＜ 0.001;t =4.289P ＜ 0.001,t =5.666,P ＜ 0.001).Compared with the control group,the level of CD3 + [(53.45 ± 6.54) ％ vs.(50.36±±3.74)％],CD4+[(24.09±±2.91)％ vs.(21.26±2.37)％] andCD4+/CD8+(1.19±0.31vs.0.96 ± 0.26) of the patients of the observation group after the treatment were significantly improved,and CD8 +[(27.87 ± 3.26) ％ vs.(31.64 ± 4.05) ％] was significantly decreased,all with statistically significant differences (t=3.407,P=0.001;t =6.264,P＜0.001;t =4.722,P＜0.001;t =6.023,P＜0.001).Compared with the control group,the total adverse reaction rate of the observation group was significantly decreased (36.23％ vs.55.07％),with a statistically significant difference (x2 =4.936,P =0.026).Conclusion Dingkoulizhong decoction can significantly improve the clinical efficacy in patients with advanced gastric cancer treated with chemotherapy,alleviate the immune function damage caused by chemotherapy,and it can reduce the occurrence of adverse reactions.
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<p><b>OBJECTIVE</b>To investigate the effects of JAZF1 overexpression on the pro-inflammatory cytokines in hepatic steatosis.</p><p><b>METHODS</b>The model of hepatic steatosis was established by incubating hepatocytes with palmitic acid (PA) at 0, 0.125, 0.25, 0.5 and 1 mM dose and for 0, 6, 12, 24 and 48 hours, after which recombinant adenovirus expressing JAZF1 (Ad-JAZF1) was introduced to up-regulate expression. Triglyceride level was measured by GOD. Cell viability was detected by CCK-8. The mRNA and protein expression of TNF-alpha, MCP-1, IL-8 and JAZF1 was examined by RT-PCR, ELISA, and western blotting.</p><p><b>RESULTS</b>The PA-treated hepatocytes showed dose-dependent significant increases in TNF-alpha, MCP-1 and IL-8 mRNA expression for doses up to 0.25 mM; there were no significant increases for the highest doses of 0.5 and 1 mM. The 0.25 mM PA-treated hepatocytes showed time-dependent significant increases in TNF-alpha, MCP-1 and IL-8 mRNA expressions (FTNF-alpha = 26.51, FMCP-1 = 57.20, FIL-8 = 353.85, P less than 0.01), with the maximum level reached at 12 h and followed by a gradual decrease with longer treatment times. JAZF1 mRNA and protein expression was markedly increased in hepatocytes infected with Ad-JAZF1 (P less than 0.01). However, the AP-treated hepatocytes with JAZF1 overexpression showed down-regulation of TNF-alpha, MCP-1 and IL-8 mRNA expression (decreased by 89.69%, 77.68%, and 83.21%, respectively) and secretion (37%, 37% and 41%, respectively, P less than 0.01).</p><p><b>CONCLUSION</b>Stimulation of hepatocytes by the PA fatty acid in vitro promotes mRNA expression of TNF-alpha, MCP-1 and IL-8, but overexpression of JAZF1 inhibits the PA-induced expression and secretion of these factors.</p>
Subject(s)
Humans , Cell Survival , Chemokine CCL2 , Metabolism , Cytokines , Metabolism , Fatty Liver , Hepatocytes , Metabolism , Interleukin-8 , Metabolism , Neoplasm Proteins , Metabolism , Palmitic Acid , Pharmacology , RNA, Messenger , Metabolism , Tumor Necrosis Factor-alpha , Metabolism , Up-RegulationABSTRACT
cry1Ah1, one of holo-type cry genes, cloned in this laboratory from Bacillus thuringiensis strain has been patented in China, and it encoded a protein with strong insecticidal activity against certain lepidopteran insect pests, such as Chilo suppressalis. cry1Ah1 gene is exhibiting good application prospects. In order to improve the expression level of cry1Ah1 gene in rice, and investigate the effect of codon usage preference of gene expression, we designed five different optimized schemes for cry1Ah1 insecticidal critical fragment in accordance with bias of rice codon, to improve G+C content, removed the shear signal and unstable factors. Optimized cry1Ah1 genes were transformed into Escherichia coli Rosetta (DE3) respectively, and 65 kDa polypeptides was expressed normally in inclusion body separately. All of these expressed polypeptides showed insecticidal activity against 2nd-instar larvae of Plutella xylostella and neonate of Chilo suppressalis. After transformation with modified cry1Ah1 genes into Var nippobare, the transgenic rice seedlings were detected by PCR, the positive rate containing target gene was more than 87%. Afterwards, the results of real-time RT-PCR and ELISA assay indicated that the highest expression level of five modified cry1Ah1 genes was that using the highest frequent codons. Average expression amount of Cry1Ah1 polypeptides was 0.104% of total soluble proteins from the positive transgenic rice.
