Role of post-translational modification of basic leucine zipper transcription factors in response to abiotic stresses in plants / 生物工程学报

Abiotic stresses substantially affect the growth and development of plants. Plants have evolved multiple strategies to cope with the environmental stresses, among which transcription factors play an important role in regulating the tolerance to abiotic stresses. Basic leucine zipper transcription factors (bZIP) are one of the largest gene families. The stability and activity of bZIP transcription factors could be regulated by different post-translational modifications (PTMs) in response to various intracellular or extracellular stresses. This paper introduces the structural feature and classification of bZIP transcription factors, followed by summarizing the PTMs of bZIP transcription factors, such as phosphorylation, ubiquitination and small ubiquitin-like modifier (SUMO) modification, in response to abiotic stresses. In addition, future perspectives were prospected, which may facilitate cultivating excellent stress-resistant crop varieties by regulating the PTMs of bZIP transcription factors.

Research progress of risk factors of Leber’s hereditary optic neuropathy / 中华眼底病杂志

Leber’s hereditary optic neuropathy (LHON) is a paradigm maternal hereditary eye disease, mainly involving the retinal and macular fibers of the optic disc in the anterior ethmoid plate of the sclera. LHON has the characteristics of sex bias among males and incomplete penetrance. Primary mitochondrial DNA mutations m.11778G>A, m. 14484T>C, m.3460G>A are the molecular basis of LHON. However, other risk factors, such as secondary mitochondrial DNA mutations, mitochondrial haplotypes, nuclear modification genes, estrogen, vitamin B12 and environmental factors, work together to affect its phenotypic expression. The clinical diagnosis of LHON mainly limited to the detection of the primary mutation site of mitochondrial DNA. Therefore, comprehensive analysis of multiple risk factors of LHON will facilitate to construct multi-dimensional model of prevention, diagnosis and treatment system, which provide accurate and individualized medical services for patients. These may alleviate the incidence in LHON families. It also provides new ideas and different angles for the in-depth study of the pathogenesis of LHON.

Role of PIAS-regulated SUMOylation of PPARγ in endogenous protective mechanism against endotoxin-induced acute lung injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of small ubiquitin-associated modifier (SUMO) E3 ligase (PIAS)-regulated SUMOylation of peroxisome proliferator-activated receptor γ (PPARγ) in the endogenous protective mechanism against endotoxin-induced acute lung injury (ALI) in mice.Methods:Experiment Ⅰ Twenty-four clean-grade wild type male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (C group), ALI group, ALI+ PPARγ inducer TZD group (ALI+ T group) and ALI+ TZD+ SUMOylation inhibitor anacardic acid group (ALI+ T+ A group). Lipopolysaccharide (LPS) 15 mg/kg was injected into the tail vein to develop the ALI model. In ALI+ T+ A group, anacardic acid 5 mg/kg was intraperitoneally injected at 1 h before LPS administration. In ALI+ T group and ALI+ T+ A group, TZD 50 mg/kg was intraperitoneally injected at 30 min before LPS administration. The mice were sacrificed at 12 h after LPS administration, and the lung tissues were obtained to examine the pathological changes which were scored and to determine the wet/dry (W/D) weight ratio, and expression of PIAS1, PIAS2, PIAS3 and PIASy protein and mRNA (by Western blot or polymerase chain reaction). Experiment Ⅱ Mouse alveolar macrophages (MH-S cells) were cultured in vitro and divided into 4 groups ( n=5 each) using a random number table method: control group (C group), LPS group, LPS+ PIAS2 siRNA group (L+ P group) and LPS+ Con siRNA group (L+ C group). Cells were routinely cultured in group C. Cells were stimulated with 10 μg/ml LPS to develop the model of endotoxin challenge. PIAS2 siRNA 50 nmol/L and Con siRNA 50 nmol/L were transfected at 48 h before LPS was added in L+ P group and L+ C group, respectively. The cells were collected at 24 h of incubation with LPS to determine the cell viability, levels of M1 and M2 alveolar macrophages (by flow cytometry), expression of PIAS2 and PPARγ (by Western blot), co-expression of PPARγ-SUMO1 (by immunoprecipitation) and expression of tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) mRNA (by polymerase chain reaction). The ratio of M1/M2 was calculated. Results:Experiment Ⅰ Compared with C group, the lung injury scores and W/D ratio were significantly increased, and the expression of PIAS2 protein and mRNA was up-regulated in the other three groups ( P<0.05). Compared with ALI group, the lung injury scores and W/D ratio were significantly decreased, and the expression of PIAS2 protein and mRNA was up-regulated in ALI+ T group and ALI+ T+ A group ( P<0.05). Compared with ALI+ T group, the lung injury scores and W/D ratio were significantly increased, and the expression of PIAS2 protein and mRNA was down-regulated in ALI+ T+ A group ( P<0.05). There was no significant difference in the expression of PIAS1, PIAS3 and PIASy protein and mRNA in lung tissues among the four groups ( P>0.05). Experiment Ⅱ Compared with C group, the cell viability was significantly decreased, the expression of PPARγ and co-expression of PPARγ-SUMO1 was up-regulated, the levels of M1 and M2 macrophages and M1/M2 ratio were increased, the expression of TNF-α mRNA was up-regulated, and the expression of IL-10 mRNA was down-regulated in the other three groups, and PIAS2 expression was significantly up-regulated in L group and L+ C group ( P<0.05). Compared with L group, the cell viability was significantly decreased, the expression of PIAS2 and PPARγ and PPARγ-SUMO1 co-expression were down-regulated, the M1 macrophage level and M1/M2 ratio were increased, TNF-α mRNA expression was up-regulated, and the expression of IL-10 mRNA was down-regulated in L+ P group ( P<0.05), and no significant change was found in the parameters mentioned above in L+ C group ( P>0.05). Compared with L+ C group, the cell viability was significantly decreased, the expression of PIAS2 and PPARγ and co-expression of PPARγ-SUMO1 were down-regulated, the level of M1 alveolar macrophages and M1/M2 ratio were increased, the expression of TNF-α mRNA was down-regulated, and the expression of IL-10 mRNA was up-regulated in L+ P group ( P<0.05). Conclusions:PIAS2-regulated SUMOylation of PPARγ is the endogenous protective mechanism against endotoxin-induced ALI in mice, which may be related to inhibition of macrophage polarization into M1 type and alleviation of inflammatory responses.

Effect of selective cerebral mild hypothermia on SUMO2/3 modification of Drp1 in a rat model of cerebral ischemia-reperfusion / 中华麻醉学杂志

Objective:To evaluate the effect of selective cerebral mild hypothermia on small ubiquitin-like modifier 2/3 (SUMO2/3) modification of dynamin-related protein 1 (Drp1) in a rat model of cerebral ischemia-reperfusion (I/R).Methods:Sixty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 240-260 g, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (S group), cerebral I/R group (I/R group), selective cerebral mild hypothermia group (HT group) and normal temperature group (NT group). The operation was performed under the monitoring of cerebral temperature and rectal temperature.Only the cervical blood vessels were exposed in S group, while focal cerebral I/R was induced by 2 h middle cerebral artery occlusion (MCAO) followed by 24 h reperfusion in anesthetized animals in the other three groups.In HT group and NT group, 4 and 37 ℃ normal saline was perfused through the left internal carotid artery at a rate of 80 ml·kg -1·h -1 for 15 min, respectively. Modified neurological severity score (mNSS) was assessed at 24 h of reperfusion. Then the rats were sacrificed under deep anesthesia, brains were removed, brain tissues were obtained for determination of the percentage of cerebral infarct size (by TTC staining), and the ischemic penumbra tissues in the cerebral cortex were removed for examination of the ultra-structural changes of mitochondria (with a transmission electron microscope) and for determination of the SUMO2/3 modification of Drp1 (by CO-IP), expression of total Drp1 (T-Drp1) and total cytochrome c (T-Cytc) (by Western blot), and expression of mitochondrial outer membrane Drp1 (M-Drp1) and cytoplasmic Cytc (C-Cytc) (by Western blot) after isolation of mitochondria and cytoplasm. Results:Compared with S group, the mNSS and percentage of cerebral infarct size were significantly increased, the expression of M-Drp1, T-Drp1, C-Cytc and T-Cytc was up-regulated, and SUMO2/3 modification of Drp1 in ischemic penumbra area was increased ( P<0.05), the fragmentation of mitochondria was aggravated, and cristae rupture and vacuolation were obvious in the other three groups. Compared with I/R group, the mNSS and percentage of cerebral infarct size were significantly decreased, the expression of M-Drp1, T-Drp1, C-Cytc and T-Cytc was down-regulated, SUMO2/3 modification of Drp1 was increased ( P<0.05), the fragmentation of mitochondria was significantly attenuated, and cristae rupture and vacuolation were weakened in HT group. There were no significant differences in these detection parameters between NT group and I/R group ( P>0.05). Conclusions:The mechanism by which selective cerebral mild hypothermia alleviates the cerebral I/R injury is related to increased SUMO2/3 modification of Drp1, decreased binding of Drp1 to mitochondrial outer membrane, and reduced mitochondrial excessive fission in rats.

Breath-by-breath measurement of exhaled ammonia by acetone-modifier positive photoionization ion mobility spectrometry via online dilution and purging sampling / 药物分析学报

Exhaled ammonia(NH3)is an essential noninvasive biomarker for disease diagnosis.In this study,an acetone-modifier positive photoionization ion mobility spectrometry(AM-PIMS)method was developed for accurate qualitative and quantitative analysis of exhaled NH3 with high selectivity and sensitivity.Acetone was introduced into the drift tube along with the drift gas as a modifier,and the characteristic NH3 product ion peak of(C3H6O)4NH4+(K0=1.45 cm2/V·s)was obtained through the ion-molecule reaction with acetone reactant ions(C3H6O)2H+(K0=1.87 cm2/V·s),which significantly increased the peak-to-peak resolution and improved the accuracy of exhaled NH3 qualitative identification.Moreover,the interference of high humidity and the memory effect of NH3 molecules were significantly reduced via online dilution and purging sampling,thus realizing breath-by-breath measurement.As a result,a wide quantitative range of 5.87-140.92 μmol/L with a response time of 40 ms was achieved,and the exhaled NH3 profile could be synchronized with the concentration curve of exhaled CO2.Finally,the analytical capacity of AM-PIMS was demonstrated by measuring the exhaled NH3 of healthy subjects,demon-strating its great potential for clinical disease diagnosis.

The pharmacological effects of isatin on Parkinson's disease / 中国药理学通报

Parkinson's disease (PD) is a kind of senile neurodegenerative disease. Dopaminergic drugs and anticholinergic drugs are the two major therapeutic drugs for PD. In the past several decades, great progress has been achieved on dopamines (DAs) and their synergists including monoamine oxidase B (MAO-B) inhibitors, catechol oxygen methyl transferase inhibitors, ergot and nonergot DA receptor agonists, DA precursor drugs, cannabis and isatin. Isatin is the inhibitor of endogenous specific anti-aging enzyme MAO-B, which has a variety of pharmacological activities. Moreover, the pharmacological mechanism of isatin may be associated with the regulatory functions of various protein activities.

Anthraquinone modifier KA-4c triggers endoplasmic reticulum stress and targets ATF6 against breast cancer cells / 中国药理学通报

Aim To explore the mechanism of the effect of anthraquinone modifier KA-4c on breast cancer cells, and determine its action target by drug affinity reaction target stability technique (DARTS). Methods The cell viability was detected by MTT method. The effect of KA-4c on the morphology of breast cancer cells was studied by HE staining, ER-Tracker Red and electron microscope. The apoptosis rate of breast cancer cells induced by KA-4c was detected by flow cytometry. The expression of apoptotic protein was detected by Western blotting. DARTS and CETSA were used to determine the target of KA-4c. Results KA-4c had the most significant inhibitory effect on the proliferation of triple negative breast cancer MDA-MB231 cells, and could cause endoplasmic reticulum and mitochondrial vacuolation to damage the cells. The apoptosis rate and the expression of apoptosis-related proteins CHOP and caspase-7 increased with the increase of KA-4c concentration. DARTS results showed that KA-4c could activate endoplasmic reticulum protein processing signaling pathway, in which KA-4c bound to ATF6 protein and was resistant to protease hydrolysis. The results of CETSA experiments showed that KA-4c could enhance the expression of ATF6 protein in a concentration-dependent manner. Conclusions KA-4 triggers endoplasmic reticulum stress to induce apoptosis in breast cancer cells. ATF6 may be one of the targets of KA-4c.

SENP1 induced protein deSUMO modification increased the chemotherapy sensitivity of endometrial cancer side population cells / 中华肿瘤杂志

Objective: To inhibit the stemness maintenance potential of endometrial cancer and increase the sensitivity of endometrial cancer side population cells to chemotherapy drugs by inducing extensive deSUMOylation modification of proteins. Methods: Flow cytometry was used to sort and culture CD133(+) CD44(+) KLE endometrial cancer cell clone spheres. Protein expression level of small ubiquitin-related modifier 1 (SUMO1) and two stemness maintenance genes of tumor side population cells, octamer binding transcription factor-4 (Oct4) and sex determining region Y-box2 (Sox2), were detected by western blotting method. Lentivirus-mediated Sentrin/SUMO-specific proteases 1 (SENP1) gene was stably transfected into KLE side population cells. Western blotting was used to detect the protein expressions of SENP1, SUMO1, Oct4 and Sox2. The clone formation rate was compared between KLE side population cells with or without SENP1 overexpression. Flow cytometry was applied to detect cell cycle changes. 3-(4, 5-Dimethylthiazole-2)-2, 5-diphenyl-tetrazolium bromide (MTT) experiment and flow cytometry apoptosis method were used to detect the chemosensitivity of the side population of endometrial cancer cells to cisplatin. Tumor-bearing mouse models of endometrial cancer were established to detect the effect of SENP1 overexpression on the chemotherapy sensitivity of cisplatin. Results: Compared with CD133(-)CD44(-) KLE cells, CD133(+) CD44(+) KLE side population cells could form clonal spheres and express higher levels of SUMO1, Oct4 and Sox2 proteins (P<0.05). Compared with KLE side population cells that were not transfected with SENP1 gene, the expression level of SENP1 protein in KLE side population cells overexpressing SUMO1、Oct4 and Sox2 were lower. The clonal sphere formation rate was reduced from (25.67±5.44)% to (7.46±1.42)%, and cell cycle shifted from G(0)/G(1) phase to G(2) phase. IC(50) of cisplatin decreased from (55.46±6.14) μg/ml to (11.55±3.12) μg/ml, and cell apoptosis rate increased from (9.76±2.09)% to (16.79±3.44)%. Overexpression of SENP1 could reduce the tumorigenesis rate of KLE side population cells in vivo and increase their chemotherapy sensitivity to cisplatin (P<0.05). Conclusion: Overexpression of SENP1 can induce protein deSUMOylation modification, inhibit the stemness maintenance potential of endometrial cancer side population cells, and enhance their chemotherapy sensitivity, which provides a new reference for gene therapy of endometrial cancer.

Relationship between heatwave and years of life lost associated with stroke in Guangdong Province: Based on Bayesian spatio-temporal model / 环境与职业医学

Background Stroke has become a main cause of death in China. With global warming, the studies on temperature and stroke have attracted much attention. Objective To analyze he relationships between heatwave and the years of life lost (YLL) by different subtypes of stroke by controlling temporal and spatial effects with Bayesian spatio-temporal model, and to study the modifiers of the health effect of heatwave. Methods The daily information of stroke deaths, meteorological data, and air pollutant data in 40 districts and counties of Guangdong Province were collected during the warm seasons (from May to October) in the years from 2014 to 2017. The individual YLL was first calculated by matching age and gender according to the life table, and then the daily YLL rate (person-years/100 000 people) was obtained by summarizing the daily YLL and correcting it with the population of each district or county. Bayesian spatio-temporal model was used to fit a proposed exposure-response relationship between heatwave and the YLL rates of different subtypes of stroke. Finally, stratified analyses were conducted by age (<65 years, ≥65 years), gender (male, female), and region (Pearl River Delta and non-Pearl River Delta regions) to identify the major modifiers for the association between heatwave and stroke mortality. Results During the warm seasons from 2014 to 2017, a total of 23 heatwave events occurred in the 40 districts or counties of Guangdong Province, cumulatively lasting for 145 d. A total of 30 852 stroke deaths were recorded in the same time periods. The average daily YLL rate of total stroke was (2.39±3.63) person-years/100 000 people, and those for hemorrhagic stroke and ischemic stroke were (1.54±2.99) person-years/100 000 people and (0.84±1.85) person-years/100 000 people, respectively. Heatwave was associated with increased YLL rate of stroke in residents, and it had a greater impact on ischemic stroke with a lag effect. The largest cumulative effect of heatwave was at lag 0-1 day, which was associated with an increased YLL rate of total stroke and ischemic stroke by 0.17 (95%CI: 0.03-0.29) person-years/100 000 people and 0.13 (95%CI: 0.06-0.20) person-years/100 000 people, respectively. The results of stratified analyses showed that heatwave had a larger effect on ischemic stroke in residents of aged 65 years or older, male, and non-Pearl River Delta regions, and the rates of YLL increased by 1.11 (95%CI: 0.58-1.55), 0.13 (95%CI: 0.03-0.23), and 0.20 (95%CI: 0.07-0.32) person-years/100 000 people, respectively; Heatwave only had an effect on hemorrhagic stroke in residents aged 65 years or older with an increased YLL rate of 0.79 (95%CI: 0.26-1.31) person-years/100 000 people. Conclusion Heatwave could elevate the level of years of life lost associated with stroke in Guangdong residents, with greater impacts on ischemic stroke of the aged, men, and residents in non-Pearl River Delta regions, and on hemorrhagic stroke in the elderly.

Clinical value of deubiquitination modification in evaluating the prognosis of patients with diffuse large B-cell lymphoma / 中国医师进修杂志

Objective:To observe the expression levels of small ubiquitinated protein specific protease (SENP) 1 and small ubiquitin-related modifier protein (SUMO) 1 in patients with diffuse large B-cell lymphoma (DLBCL), and analyze the clinical value of evaluating prognosis.Methods:The clinical data of 66 patients with DLBCL (DLBCL group) in Inner Mongolia People′s Hospital from February 2017 to October 2020 were retrospectively analyzed, and 60 cases of healthy people in the same period were selected as the healthy control group. The expression levels of SENP1 and SUMO1 were detected by enzyme linked immunosorbent assay (ELISA). The correlation between the expression levels of SENP1, SUMO1 and clinical characteristics was analyzed. The independent risk factors affecting the prognosis were analyzed by Cox multivariate analysis.Results:The SENP1 in DLBCL group was significantly higher than that in healthy control group (50.39 ± 6.86 vs. 7.47 ± 1.32), the SUMO1 in DLBCL group was significantly lower than that in healthy control group (8.84 ± 2.13 vs. 31.49 ± 5.89), and there were statistical differences ( t = 47.640 and 29.210, P<0.01). There were statistical differences in SENP1 and SUMO1 among patients with different clinical stages ( P<0.01). The expression levels of SENP1 and SUMO1 were correlated with clinical stage and international prognostic index (IPI) ( P<0.05), and were not correlated with age, gender, disease site and clinical symptoms ( P>0.05). The 3-year survival rate in patients with high SENP1 expression (30 cases) was significantly lower than that in patients with low SENP1 expression (36 cases), the 3-year survival rate in patients with high SUMO1 expression (38 cases) was significantly higher than that in patients with low SUMO1 expression (28 cases), and there were statistical differences (26.67% vs. 75.00% and 73.68% vs. 39.29%, P<0.05). Cox multivariate regression analysis result showed that clinical stage, IPI, SENP1 and SUMO1 were independent risk factors affecting the prognosis in patients with DLBCL ( HR = 1.352, 1.487, 2.048 and 3.295; 95% CI 1.180 to 1.691, 1.187 to 1.602, 2.536 to 4.023 and 2.752 to 5.325; P<0.05 or <0.01). Conclusions:In patients with DLBCL, SENP1 is highly expressed and SUMO1 is lowly expressed. The expression levels of SENP1 and SUMO1 are closely related to clinical stage and IPI in patients with DLBCL, and they are independent risk factors of the prognosis.

Effect of small ubiquitin-like modifier 1 pseudogene 3 on the proliferation and apoptosis of non-small cell lung cancer cell line H1299 through protein kinase B signaling pathway / 解剖学报

Objective To investigate the effect of long noncoding RNA (IncRNA) small ubiquitin-like modifier 1 pseudogene 3(SUM01P3) on the proliferation and apoptosis of non-small cell lung cancer cell line 1299. Methods Determination of SUM01P3 expression in non-small cell lung cancer cells by Real-time PCR. SUM01P3 small interfering RNA(siRNA) was transfected into H1299 cells, the down regulation effect was determined by Real-time PCR. Cell proliferation was measured by MTT, 5-ethynyl-2′-deoxyuridine(EdU) method, the cell cycle was determined by PI single staining, apoptosis was detected by annexin V -FITC/PI, detection of apoptosis by TUNEL, Western blotting was used to detect the expression of cleaved Caspase-3 (c-Caspase-3), cyclin Dl, P27, phosphorylated phospoinositide 3-kinase (p-PI3K) and phosphorylated protein kinase B (p-Akt). Akt signal activator treated H1299 cells transfected with SUM01P3 siRNA, cell proliferation, apoptosis and cycle change were also measured by the above methods. The number of samples was 9. Results SUM01P3 was up-regulated in non-small cell lung cancer cells. The expression of SUM01P3 in H1299 cells decreased after transfection with SUM01P3 siRNA, cell proliferation decreased, the ratio of G0/Gj phase increased, apoptosis rate increased, c-Caspase-3 and P27 protein in the cells increased, the protein levels of cyclin D1, p-PI3K and p-Akt decreased. Akt signal activator could reverse the inhibition of proliferation, cycle arrest and apoptosis of H1299 cells by SUM01P3 siRNA. Conclusion Down-regulation of SUM01P3 inhibits the proliferation of non-small cell lung cancer H1299 cells and induces apoptosis, the mechanism of action is related to the reduction of the activation level of the Akt signaling pathway.

Small ubiquitin-like modifier proteins specific protease 3 in microglia participating in the early progression of photothrombosic ischemic stroke mice model / 解剖学报

Objective To investigate the expression of small ubiquitin-like modifier proteins specific protease 3 (SENP3) in microglia of mice with ischemic stroke and its relationship with the progression of ischemic stroke. Methods The experimental animals were divided into control group, ischemic stroke day 1 group and ischemic stroke day 7 group (3 mice per group). The expression of inducible nitric oxide synthase (iNOS), argniase-1 (ARG-1), SENP3 and c-Jun N-terminal kinase (JNK) phosphorylation levels in the striatum were detected by immunoblotting. The expression of iNOS and ARG-1 in mouse striatum microglia was detected by immunofluorescence double labeling. Results Compared with the control group, the expression of SENP3 and the phosphorylation level of JNK in the ischemic stroke group increased significantly, and the expression of the marker iNOS of Ml type microglia increased significantly. The expression of the marker ARG-1 of M2 type microglia increased significantly in the day 7 group of ischemic stroke. The immunofluorescence double-labeled result of striatum ionized calcium binding adapter molecule 1 (Ibal) and iNOS, Ibal and ARG-1 were consistent with the result of immunoblotting. Conclusion In the early stage of ischemic stroke, the expression of SENP3 in microglia increases, which promote the cerebral inflammatory response by affecting the level of JNK phosphorylation and the polarization of microglia, and participate in the progression of ischemic stroke.

Desenvolvimento de achocolatados em pó com adição de subprodutos de frutas, processados por spray dryer e com modificador reológico / Development of powdered chocolate added with fruit byproducts, processed by spray dryer and rheological modifier

O achocolatado em pó é um dos derivados do cacau com maior inserção econômica e cultural em diversos países. O objetivo deste estudo foi avaliar a adição de ingredientes diferenciados nesse tipo de produto, como modificadores reológicos e fruta, bem como à alteração no tipo de cacau utilizado, ocasionando mudanças sensoriais e nutricionais positivas ao produto. O fruto sugerido neste estudo foi o cupuaçu (Theoboroma grandiflorum), fruto típico da região Norte, que apresenta excelente qualidade nutricional. Foram desenvolvidas 7 formulações de achocolatado por método convencional após simples mistura (padrão, com cacau alcalino, com cacau orgânico, com polpa de cupuaçu, com amido pré-gelatinizado, com amido pré-gelatinizado + polpa de cupuaçu, com goma guar, com goma guar + polpa de cupuaçu) e 4 formulações processadas por spray dryer após a simples mistura (padrão, com polpa de cupuaçu, com amido pré-gelatinizado, com amido + polpa de cupuaçu). Todas as formulações foram avaliadas quanto à composição nutricional, calorimetria exploratória de varredura (DSC), análises físico-químicas, reológica, quantificação dos compostos fenólicos e avaliação da capacidade antioxidante por métodos in vitro. Em seguida foi realizada análise sensorial com as formulações: padrão, com polpa de cupuaçu e com amido + polpa de cupuaçu. O achocolatado padrão apresentou tempo de mistura de 38 min, o que foi utilizado como parâmetro para as demais formulações. Os achocolatados que continham polpa de cupuaçu apresentaram maior teor proteico (14,5 a 16,3 g/100g) quando comparados com o padrão (13,6 g/100g). Todos os achocolatados apresentaram umidade entre 1,2% e 3,7%, e atividade de água entre 0,13 e 0,57, considerados microbiologicamente estáveis, sendo bom para a vida útil do produto. Os achocolatados obtiveram tempo de molhabilidade entre 07:15 min e 15:06 min; solubilidade de 1,56 IR% a 7,44 IR%; tamanho de partícula variando entre 0,216 mm e 0,347 mm (partículas finas). O uso do spray dryer não teve impacto significativo nas características físicas das formulações, assim como a utilização dos diferentes tipos de cacau não afetou a composição nutricional e qualidade física dos achocolatados. Houve aumento (p< 0,05) para o tempo de molhabilidade e solubilidade do achocolatado com cacau orgânico em comparação com o padrão (13:30 e 9:33 min; 2,64 e 1,56 IR%, respectivamente). A transição vítrea variou entre 35,2 a 35,7 mW enquanto o ponto de carbonização ficou entre 237,4 a 243,6 mW, indicando que a adição dos agentes espessantes e/ou do cupuaçu não interferiu (p<0,05) na análise térmica dos achocolatados. Todos os achocolatados diluídos em leite apresentaram-se como pseudoplásticos, com aumento de viscosidade nas menores temperaturas, conforme esperado. O achocolatado com cacau orgânico apresentou o maior teor de compostos fenólicos (8,27 mg AG g-1) enquanto observou-se redução no conteúdo de fenólicos nos produtos processados por spray dryer. Os achocolatados apresentaram capacidade antioxidante entre 31,76 µMETrolox/g e 75,62 µMETrolox/g, pelo método do DPPH. A adição do cupuaçu levou ao aumento da capacidade de sequestro de radicais DPPH quando comprados com o padrão (p<0,05). Não foi observada diferença significativa pelo método FRAP. A avaliação sensorial obteve aceitação situada na região positiva da escala (5 a 7). Os achocolatados formulados apresentam formulações adequadas a sua comercialização, com agregação de valor nutricional e econômico

The chocolate powdered is a cocoa-derived with greater economic and cultural integration in several countries. The objective of this study was to evaluate the addition of different ingredients in this type of product, such as rheological modifiers and fruit, as well as the change in the type of cocoa used, causing positive sensory and nutritional changes to the product. The fruit suggested in this study was cupuassu (Theoboroma grandiflorum), a typical fruit from the northern region, which has excellent nutritional quality. Seven powdered chocolate formulations were developed by conventional method after simple mixing (standard, with alkaline cocoa, with organic cocoa, with cupuassu pulp, pre-gelatinized starch, pre-gelatinized starch + cupuassu pulp, guar gum, with guar gum + cupuassu pulp) and 4 formulations processed by spray dryer after simple mixing (standard, with cupuassu pulp, pre-gelatinized starch, starch + cupuassu pulp). All formulations were evaluated for nutritional composition, differential scanning calorimetry (DSC), physicochemical, rheological analyzes, quantification of phenolic compounds and antioxidant capacity evaluation by in vitro methods. Then, sensory analysis was performed with the formulations: standard, with cupuassu pulp and starch + cupuassu pulp. The standard powdered chocolate had a mixing time of 38 min, which was used as parameter for the other formulations. The powdered chocolate containing cupuassu pulp had higher protein content (14.5 to 16.3 g / 100g) when compared to the standard (13.6 g / 100g). All powdered chocolate presented humidity between 1.2% and 3.7%, and water activity between 0.13 and 0.57, considered microbiologically stable, wich is good for the shelf life of the product. The powdered chocolate obtained wettability time between 07:15 min and 15:06 min; solubility from 1.56 IR% to 7.44 IR%; particle size ranging from 0.216 mm to 0.347 mm (fine particles). The use of the spray dryer had no significant impact on the physical characteristics of the formulations, as well as the use of different types of cocoa did not affect the nutritional composition and physical quality of the powdered chocolate. There was an increase (p <0.05) for the time of wettability and solubility in chocolate powdered formulated with organic cocoa when compared to the standard (9:33 and 13:30 min; IR 2.64 and 1.56%, respectively). The glass transition ranged from 35.2 to 35.7 mW while the carbonization point ranged from 237.4 to 243.6 mW, indicating that the addition of thickening agents and / or cupuassu did not interfere (p <0.05) in the thermal analysis of powdered chocolate. All powdered chocolate when diluted in milk presented as pseudoplastics, with viscosity increase at lower temperatures, as expected. Chocolate powdered with organic cocoa presented the highest content of phenolic compounds (8.27 mg AG g-1) whereas there was a reduction in phenolic content in products processed by spray dryer. The powdered chocolates presented antioxidant capacity between 31.76 µMETrolox / g and 75.62 µMETrolox / g, by the DPPH method. The addition of cupuassu led to increased ability to sequester DPPH radicals when compared to the standard (p <0.05). No significant difference was observed by the FRAP method. Sensory evaluation was accepted in the positive region of the scale (5 to 7). The formulated powdered chocolates have appropriate formulations for marketing, with added nutritional and economic value

Epigenetic regulation of hepatitis B virus cccDNA and related prospects for antiviral therapy / 临床肝胆病杂志

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is stably maintained in hepatocytes in the form of minichromosome and is considered the most important cause of chronicity of HBV infection, presence of HBV after antiviral therapy, and recurrence of hepatitis after drug withdrawal. However, due to a lack of antiviral regimens targeting cccDNA itself or the formation and transcription of cccDNA, there is an urgent need for treatment strategies targeting cccDNA. With the gradual understanding of epigenetic modification of histones of the cccDNA minichromosome, epigenetic therapy is expected to become a potential therapy for HBV. This article reviews the current status and future directions of HBV DNA methylation and cccDNA-bound histone modification, in order to provide new thoughts for epigenetic therapy for HBV.

The value of anti-SUMO antibody in the clinical diagnosis and treatment of primary biliary cholangitis / 国际检验医学杂志

Objective In this study, we constructed a plasmid, specifically expressed SUMO protein, and to study the expression level of anti-SUMO antibody in the serum of patients with primary biliary cholangitis, systemic lupus erythematosus, rheumatoid arthritis, Connective tissue disease, sjogren′s syndrome which were typical of autoimmune diseases.And to investigate whether anti-SUMO antibodies can be used as specific serum markers of PBC.Methods Plasmids containing SUMO1, SUMO2 and SUMO3 fragments were prepared by PCR and ET cloning and introduced into E.coli for protein induction and purification, respectively.Dot blot was used to preliminarily screen anti-SUMO antibody-positive specimens from serum samples of PBC patients and were verified by western blot to obtain positive reference serum.Through the establishment of the optimal anti-SUMO antibody ELISA diagnostic system, the positive rates of three subtypes of anti-SUMO antibody in PBC, SLE, SS, RA and CTD were detected, and their differences were analyzed by chi-square test.ResultsThe anti-SUMO antibody label has a specificity of up to 99％in PBC and a sensitivity of around 86％.After chi-square test analysis, the positive detection rate of anti-SUMO antibody in PBC was higher than that of nonPBC autoimmune disease and healthy controls (P＜0.01).There was no significant difference in the expression of anti-SUMO antibody in other autoimmune disease populations (non-PBC autoimmune diseases) (P＞0.05).Conclusion The highly specific anti-SUMO antibody is expected to become a novel antibody for diagnosis of PBC, which is of great significance for improving the clinical diagnosis efficiency of PBC.

Hypoxia increases chemotherapy resistance in nasopharyngeal carcinoma via inducing CDK6 deSUMOylation / 中华耳鼻咽喉头颈外科杂志

Objective@#To understand the mechanism of chemotherapy resistance in nasopharyngeal carcinoma under hypoxic conditions through the perspective of protein SUMOylation modification.@*Methods@#Cobalt chloride (CoCl2) was used to establish the hypoxic model of human nasopharyngeal carcinoma CNE1 cells. Then, the cell cycle was detected by flow cytometry, and the expression level of small ubiquitin-related modifier(SUMO) and cyclin-dependent kinase 6 (CDK6) proteins were detected by western blotting. MTT assay was used to determine the median lethal dose (IC50) of cancer cells against cisplatin, and enzyme-linked immunosorbent assay (ELISA) was used to determine lactate dehydrogenase (LDH) level.@*Results@#The cell cycle of CNE1 induced by hypoxia was arrested in G0/G1 phase.The results of Western blot showed that the protein expression level of CDK6 in CNE1 cells was lower than that in the control group (0.83±0.25 vs. 0.43±0.21, t=14.67, P=0.003). The protein level of conjugated SUMO1 was significantly lower than that in the control group (2.69±0.48 vs. 1.38±0.31, t=17.22, P=0.001), while the level of free SUMO1 protein was significantly higher than that in the control group (2.01±0.43 vs. 2.60±0.59, t=15.45, P=0.002).The LC50 of CNE1 cells in the control group was significantly lower than that in the hypoxic group (29.44 μg/ml vs. 97.72 μg/ml, t=12.79, P=0.001). After CNE1 cells received 50 μg/ml cisplatin for 48 h, the LDH content in the supernatant of the control group was significantly higher than that in the hypoxic group ((541.49±64.59) ng/ml vs. (234.67±41.03) ng/ml, t=11.94, P=0.007)). The apoptosis rate of CNE1 cells in the control group was significantly higher than that in the hypoxic group ((76.64±5.37)% vs. (32.84±4.77) ng/ml, t=8.49, P=0.003)).@*Conclusion@#Hypoxia can dissociate the covalent modification of CDK6 and SUMO1, inhibit cell cycle and increase the chemotherapy resistance of nasopharyngeal carcinoma.

Effect of selective brain hypothermia on SUMOylation of cerebral cortex during cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of selective brain hypothermia on small ubiquitin-like modifier (SUMO)ylation of cerebral cortex during cerebral ischemia-reperfusion (I/R) in rats.Methods A total of 120 healthy clean-grade male Sprague-Dawley rats,aged 8 weeks,weighing 200-250 g,were divided into 4 groups (n=30 each) by a random number table method:sham operation group (group S),group I/R,selective brain hypothermia group (group H) and 37 ℃ normal saline group (group N).Rats were anesthetized with 10％ chloral hydrate 3 ml/kg.Cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into the right internal carotid artery and advanced cranially until resistance was met.The right middle cerebral artery was occluded for 2 h followed by reperfusion in group I/R.10-13 ℃ normal saline was infused at the rate of 100 ml · kg-1 · h-1 for 20 min starting from the time point immediately after removing the nylon thread in group H.37 ℃ normal saline was infused instead at the same rate for 20 min in N group.The neurological deficit were assessed and scored at 6,24 and 48 h after reperfusion.The cerebral cortex on ischemic side was then removed for determination of cell apoptosis (by TUNEL) and expression of SUMO sepcific proteases 3 (SENP3) and SUMO2/3-conjugated protein (by Western blot).The apoptotic rate was calculated.Results Compared with group S,the neurological deficit score and apoptosis rate were significantly increased,and the expression of SUMO2/3-conjugated protein was up-regulated at each time point after reperfusion in the other three groups (P＜0.05),and the expression of SENP3 was significantly up-regulated in I/R and N groups and down-regulated in group H (P＜0.01).Compared with group I/R,the neurological deficit score and apoptosis rate were significantly decreased,the expression of SUMO2/3-conjugated protein was up-regulated,and the expression of SENP3 was down-regulated in group H (P＜0.05).Conclusion The mechanism by which selective brain hypothermia reduces cerebral I/R injury is related to enhancing SUMOylation of cerebral cortex in rats.

Roles of SUMO modification in pulmonary diseases / 国际儿科学杂志

Small ubiquitin-related modifier(SUMO) is a small molecule that is structurally similar to ubiquitin.SUMOylation has emerged as an important post-translational modification.It profoundly influences the function of proteins,including the regulation of protein-protein interaction,the activity of protein transcription,DNA damage repair,gene regulation and biological rhythm.In recent years,evidence suggests that the SUMOylation plays crucial roles in lung.By binding to target proteins,SUMO is associated with pulmonary biological function and disease.

Research progress on antitumor effect and mechanism of carboxymethyl-glucan / 中草药

D-glucan, an immune polysaccharide, is one kind of vital biological response modifiers (BRMs) which has the function of immune regulation. Carboxymethyl-glucan (CMG) is a water-soluble derivative of D-glucan that possesses excellent immunocompetent and water-solubility. Therefore, it has broad application prospects in anticancer therapy. The research progress of antitumor effect and mechanism of CMG was reviewed in this paper, which provides a scientific basis for the development and utilization of CMG as a candidate for the prevention and treatment of cancer.

IL-21 modulates biological characteristics of NK92 cells by upregulating SENP1 expression / 军事医学

Objective To investigate the roles of SENP1 in regulation of biological characteristics of NK cells.Methods Lentivirus-mediated-Senp1-small-hairpinRNA (shRNA) transduction was applied to NK92 cells.The expression of SENP1 in NK92 cells was determined by real-time PCR and Western blot.The proliferation of NK92 cells was detected by CCK-8 assay.The apoptosis of NK92 cells was determined by Annexin Ⅴ and PI labeling.The cytotoxicity of NK92 cells against K562 cells was evaluated by luciferase reporter assay.Results Treatment of NK92 cells with IL-21 resulted in SENP1 upregulation.Lentivirus mediated SENP1 knockdown reduced proliferation and increased apoptosis in NK-92 cells,but SENP1 inhibition had slight impact on the cytotoxic ability of NK92 cells to kill K562 cells.Conclusion SENP1 mediates the regulatory effect of IL-21 on the proliferation and survival of NK92 cells.