Hepatocellular carcinoma:An analysis of the expression status of stress granules and their prognostic value

BACKGROUND Hepatocellular carcinoma(HCC)is a global popular malignant tumor,which is difficult to cure,and the current treatment is limited.AIM To analyze the impacts of stress granule(SG)genes on overall survival(OS),survival time,and prognosis in HCC.METHODS The combined The Cancer Genome Atlas-Liver Hepatocellular Carcinoma(TCGA-LIHC),GSE25097,and GSE36376 datasets were utilized to obtain genetic and clinical information.Optimal hub gene numbers and corresponding coefficients were determined using the Least absolute shrinkage and selection operator model approach,and genes for constructing risk scores and corresponding correlation coefficients were calculated according to multivariate Cox regression,respectively.The prognostic model’s receiver operating characteristic(ROC)curve was produced and plotted utilizing the time ROC software package.Nomogram models were constructed to predict the outcomes at 1,3,and 5-year OS prognostications with good prediction accuracy.RESULTS We identified seven SG genes(DDX1,DKC1,BICC1,HNRNPUL1,CNOT6,DYRK3,CCDC124)having a prognostic significance and developed a risk score model.The findings of Kaplan-Meier analysis indicated that the group with a high risk exhibited significantly reduced OS in comparison with those of the low-risk group(P<0.001).The nomogram model’s findings indicate a significant enhancement in the accuracy of OS prediction for individuals with HCC in the TCGA-HCC cohort.Gene Ontology and Gene Set Enrichment Analysis suggested that these SGs might be involved in the cell cycle,RNA editing,and other biological processes.CONCLUSION Based on the impact of SG genes on HCC prognosis,in the future,it will be used as a biomarker as well as a unique therapeutic target for the identification and treatment of HCC.

Xiaotan Tongfu granules contribute to the prevention of stress ulcers

AIM:To investigate the efficacy and potential mechanism of Xiaotan Tongfu granules(XTTF)in stress ulcers.METHODS:One hundred sixty rats were randomly divided into 4 groups(n=10)as follows:the model group(MP group),the control group(CP group),the ranitidine group(RP group)and the XTTF granule group(XP group).Rats in the MP group received no drugs,rats in the CP group received 0.2 mL of a 0.9%sodium chloride solution via oral gavage,and rats in the RP and XP groups received the same volume of ranitidine(50 mg/kg)or XTTF granule(4.9 g/kg).The cold-restraint stress model was applied to induce stress ulcers after 7 consecutive days of drug administration.Afterwards,rats were sacrificed at 0,3,6 and24 h.Gastric pH was measured by a precise pH meter;gastric emptying rate(GER)was measured by using a methylcellulose test meal;myeloperoxidase activity(MPO),macrophage migration inhibitory factor(MIF),proliferating cell nuclear antigen(PCNA),and heat shock protein 70(HSP70)were measured by immunohistochemical staining;and mucosal cell apoptosis was measured by transferase dUTP nick end labeling.RESULTS:In the cold-restraint stress model,the development of stress ulcers peaked at 3 h and basically regressed after 24 h.Gastric lesions were significantly different in the RP and XP groups at each time point.Interestingly,although this index was much lower in the RP group than in the XP group immediately following stress induction(7.00±1.10 vs 10.00±1.79,P<0.05.Concerning gastric pH,between the RP and XP groups,we detected a statistically significant difference immediately after stress induction(0 h:4.56±0.47 vs 3.34±0.28,P<0.05)but not at any of the subsequent time points.For GER,compared to the RP group,GER was remarkably elevated in the XP group because a statistically significant difference was detected(3 h:46.84±2.70 vs 61.16±5.12,P<0.05;6 h:60.96±6.71 vs 73.41±6.16,P<0.05;24 h:77.47±3.17 vs 91.31±4.34,P<0.05).With respect to MPO and MIF,comparisons between the RP and XP groups revealed statistically significant differences at 3 h(MPO:18.94±1.20 vs 13.51±0.89,P<0.05;MIF:150.67±9.85 vs 122.17±5.67,P<0.05)and 6 h(MPO:13.22±1.54 vs 8.83±0.65,P<0.05;MIF:135.50±9.46 vs 109.83±6.40,P<0.05).With regard to HSP70,HSP70 expression was significantly increased in the RP and XP groups at 3 and 6 h compared to the MP and CP groups.In addition,comparing the RP and XP groups also showed statistically significant differences at 3 and 6 h.The expression of PCNA was higher in the RP and XP groups 3 h after stress induction.Between these two groups,small but statistically significant differences were observed at all of the time points(3 h:69.50±21.52 vs 79.33±15.68,P<0.05;6 h:107.83±4.40 vs 121.33±5.71,P<0.05;24 h:125.33±5.65 vs 128.50±14.49,P<0.05)except 0 h.With regard to apoptosis,the apoptotic activity in the RP and XP groups was significantly different from that in the MP and CP groups.The XP group exhibited a higher inhibition of cell apoptosis than the RP group at3 h(232.58±24.51 vs 174.46±10.35,P<0.05)and6 h(164.74±18.31 vs 117.71±12.08,P<0.05).CONCLUSION:The Xiaotan Tongfu granule was demonstrated to be similar to ranitidine in preventing stress ulcers.It exhibited multiple underlying mechanisms and deserves further study.

Stress granules inhibit endoplasmic reticulum stress-mediated apoptosis during hypoxia-induced injury in acute liver failure

BACKGROUND Stress granules(SGs)could be formed under different stimulation to inhibit cell injury.AIM To investigate whether SGs could protect hepatocytes from hypoxia-induced damage during acute liver failure(ALF)by reducing endoplasmic reticulum stress(ERS)mediated apoptosis.METHODS The agonist of SGs,arsenite(Ars)was used to intervene hypoxia-induced hepatocyte injury cellular model and ALF mice models.Further,the siRNA of activating transcription factor 4(ATF4)and SGs inhibitor anisomycin was then used to intervene in cell models.RESULTS With the increase of hypoxia time from 4 h to 12 h,the levels of HIF-1α,ERS and apoptosis gradually increased,and the expression of SGs marker G3BP1 and TIA-1 was increased and then decreased.Compared with the hypoxia cell model group and ALF mice model,the levels of HIF-1α,apoptosis and ERS were increased in the Ars intervention group.After siRNA-ATF4 intervention,the level of SGs in cells increased,and the levels of HIF-1α,ERS and apoptosis decreased.Compared with the siRNA-ATF4 group,the levels of G3BP1 in the siRNAATF4+anisomycin group were decreased,and the levels of HIF-1α,ERS and apoptosis were increased.Moreover,compared with the ALF group,the degree of liver injury and liver function,the levels of HIF-1α,ERS and apoptosis in the Ars intervention group were decreased,the level of SGs was increased.CONCLUSION SGs could protect hepatocytes from hypoxia-induced damage during ALF by reducing ERSmediated apoptosis.

Bufei Yishen granules suppress oxidative stress in rats with chronic obstructive pulmonary disease via Nrf2 signaling

OBJECTIVE To explore the antioxidant effect of Bufei Yishen granules on chronic obstructive pulmo⁃nary disease(COPD)and investigate its underlying mechanism.METHODS Forty-eight rats were randomly divided into normal,model,Bufei Yishen granules(BY)and N-acetylcysteine(NAC)groups,12 rats in each group.The stable COPD rat model was duplicated by using repeated cigarette smoke exposure combined with Klebsiella bacterial infection for 12 weeks(week 1-12),and the corresponding drugs were administered for the next 8 weeks(week 13-20).Minute volume(MV),tidal volume(TV)and peak expiratory flow(PEF)were measured by whole body plethysmography(WBP)system every 4 weeks.Before sacrificed,forced vital capacity(FVC)and forced expiratory volume 0.1(FEV0.1)were measured byPFT system.The pathological changes of lung tissue were observed by pathological techniques.Heme oxygenase 1(HO-1),superoxide dismutase 1(SOD1)and Nrf2 in lung tissue were measured by immunohisto-hemical method.The total anti oxidizing capability(T-AOC),lipid peroxide(LPO)in rat serum were measured.The expression of Nrf2,HO-1 andγ-glutamyl cysteine synthetase(γ-GCS)mRNA in lung tissue was detected by quantitative polymerase chain reac⁃tion(qPCR).The protein expression of Keap1,Nrf2 and HO-1 in lung tissue were detected by Western blotting.RESULTS①Lung function:compared with normal group,the MV in model group was significantly decreased at week 8(P<0.01),the TV and PEF were significantly decreased at week 4(P<0.01).At week 20,compared with model group,MV,TV,and PEF in the BY and NAC groups were significantly increased(P<0.01);compared with the NAC group,MV,TV,and PEF in BY group were significantly increased(P<0.01).At the end of week 20,the FVC and FEV0.1 in model group were significantly lower than that in normal group(P<0.01).Compared with model group,the FVC and FEV0.1 in the BY and NAC groups were significantly increased(P<0.05).②Oxidative indexes:Compared with Normal group,T-AOCin serum was significantly decreased in Model group,while LPO was significantly increased(P<0.01).Compared with the Model,T-AOC in BY and NAC groups was significantly increased(P<0.01),and the LPO was significantly decreased(P<0.05,P<0.01).There were no difference between the BTG and NAC.③Nrf2 signaling:Nrf2 and HO-1 in lung tissue were mainly expressed in the cytoplasm and part of the nucleus of alveolar epithelial cells.SOD1 protein was mainly distributed in bronchial epithelial cells and alveolar septa.Compared with normal group,the expression of Nrf2 in the model group was increased(P<0.01),and HO-1 and SOD1 were decreased(P<0.01).Compared with the model,the expression of Nrf2 in the BY group was significantly increased(P<0.05),and HO-1 and SOD1 in BY and NAC groups were both increased(P<0.01).Compared with the NAC group,the expression of HO-1 in BY group was increased(P<0.01).Compared with normal group,the Nrf2 mRNA expression of lung tissue in the model was significantly increased(P<0.01),the HO-1 andγ-GCS mRNA was decreased(P<0.01).Compared with model group,the Nrf2,HO-1,andγ-GCS mRNA in the BY group were increased(P<0.01),the HO-1,andγ-GCS mRNA in NAC group were increased(P<0.01).Compared with normal group,the Nrf2 protein expression of lung tissue in the model group was significantly increased(P<0.01),and HO-1 protein expression was significantly decreased(P<0.01).Compared with the model,the Nrf2 and HO-1 protein in NAC and BY groups was significantly increased(P<0.01).CONCLUSION Bufei Yishen gran⁃ules has beneficial curative effect in COPD rats,and has the same antioxidation effect as NAC,the mechanism may be involved in upregulating Nrf2 signaling.

Paclitaxel-induced stress granules increase LINE-1 mRNA stability to promote drug resistance in breast cancer cells

Abnormal expression of long interspersed element-1(LINE-1)has been implicated in drug resistance,while our previous study showed that chemotherapy drug paclitaxel(PTX)increased LINE-1 level with unknown mechanism.Bioinformatics analysis suggested the regulation of LINE-1 mRNA by drug-induced stress granules(SGs).This study aimed to explore whether and how SGs are involved in drug-induced LINE-1 increase and thereby promotes drug resistance of triple negative breast cancer(TNBC)cells.We demonstrated that SGs increased LINE-1 expression by recruiting and stabilizing LINE-1 mRNA under drug stress,thereby adapting TNBC cells to chemotherapy drugs.Moreover,LINE-1 inhibitor efavirenz(EFV)could inhibit drug-induced SG to destabilize LINE-1.Our study provides the first evidence of the regulation of LINE-1 by SGs that could be an important survival mechanism for cancer cells exposed to chemotherapy drugs.The findings provide a useful clue for developing new chemotherapeutic strategies against TNBCs.

Diverse CMT2 neuropathies are linked to aberrant G3BP interactions in stress granules

Complex diseases often involve the interplay between genetic and environmental factors. Charcot-Marie-Tooth type 2neuropathies (CMT2) are a group of genetically heterogeneous disorders, in which similar peripheral neuropathology isinexplicably caused by various mutated genes. Their possible molecular links remain elusive. Here, we found that uponenvironmental stress, many CMT2-causing mutant proteins adopt similar properties by entering stress granules (SGs), where theyaberrantly interact with G3BP and integrate into SG pathways. For example, glycyl-tRNA synthetase (GlyRS) is translocated fromthe cytoplasm into SGs upon stress, where the mutant GlyRS perturbs the G3BP-centric SG network by aberrantly binding toG3BP. This disrupts SG-mediated stress responses, leading to increased stress vulnerability in motoneurons. Disrupting thisaberrant interaction rescues SG abnormalities and alleviates motor deficits in CMT2D mice. These findings reveal a stressdependentmolecular link across diverse CMT2 mutants and provide a conceptual framework for understanding geneticheterogeneity in light of environmental stress.

芪蛭皱肺颗粒对慢性阻塞性肺疾病大鼠内质网应激的影响

目的观察芪蛭皱肺颗粒对慢性阻塞性肺疾病(COPD)大鼠内质网应激的影响,探讨其作用机制。方法采用脂多糖气管滴注+烟熏诱导COPD大鼠模型,将60只Wistar大鼠用随机数字表法分为对照组、模型组、补肺活血胶囊组和芪蛭皱肺颗粒低、中、高剂量组,每组10只。造模第29日开始,给药组分别予相应药物灌胃,对照组和模型组灌胃生理盐水,连续28 d。检测大鼠肺功能,HE染色观察肺组织形态,TUNEL染色检测肺组织细胞凋亡程度,RT-qPCR、Westernblot检测肺组织内质网应激及凋亡相关因子基因和蛋白表达。结果与对照组比较,模型组大鼠肺功能指标吸气峰流速(PIF)、呼气峰流速(PEF)、每分钟通气量(MV)显著降低,呼吸频率(F)显著升高(P<0.05);肺组织结构损伤明显,肺损伤评分及细胞凋亡率显著升高(P<0.05),肺组织葡萄糖调节蛋白78(GRP78)、蛋白激酶R样内质网激酶(PERK)、C/EBP同源蛋白(CHOP)、胱天蛋白酶(Caspase)-3、Caspase-9mRNA表达升高(P<0.05),GRP78、p-PERK、转录激活因子4(ATF4)、CHOP、Caspase-3、Caspase-9蛋白表达升高(P<0.05)。与模型组比较,芪蛭皱肺颗粒中、高剂量组及补肺活血胶囊组大鼠PIF、PEF、MV显著升高,F显著降低(P<0.05),肺组织损伤均有改善,肺损伤评分及细胞凋亡率显著降低(P<0.05),肺组织GRP78、PERK、CHOP、Caspase-3、Caspase-9 mRNA表达降低(P<0.05),GRP78、p-PERK、ATF4、CHOP、Caspase-3、Caspase-9蛋白表达降低(P<0.05)。结论芪蛭皱肺颗粒可通过抑制COPD大鼠内质网应激相关因子表达减少细胞凋亡过亢为害,从而促进未折叠蛋白反应提高内质网折叠能力,制约内质网应激状态,助其承制。

疏肝健脾颗粒对轻中度特应性皮炎合并睡眠障碍患者的疗效与抗氧化机制研究

目的观察疏肝健脾颗粒对轻中度特应性皮炎(Atopic dermatitis,AD)合并睡眠障碍(Sleep disturbances,SDs)患者的临床疗效并探讨其作用机制。方法将符合诊断与纳入标准的58例轻中度成人AD合并SDs患者随机分为治疗组与对照组,每组29例。治疗组予疏肝健脾颗粒联合除湿止痒软膏干预,对照组予盐酸西替利嗪片联合除湿止痒软膏干预,均持续4周。观测、比较两组患者治疗前后特应性皮炎指数(SCORAD)评分、失眠严重程度指数(ISI)评分及医院焦虑抑郁量表(HADS)评分差异;检测、比较血常规及肝肾功能指标、血清5-羟色胺(5-HT)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量差异。对上述评分与指标进行相关性分析。结果经4周干预,对照组总有效率为65.52%,治疗组总有效率为89.66%,显著优于对照组(P<0.05)。与干预前相比,两组的SCORAD、ISI、HADS评分与血清MDA含量均显著降低,血清5-HT及SOD含量均有升高。其中,治疗组疗后SCORAD(P<0.05)、ISI(P<0.01)评分显著低于对照组,血清SOD含量显著高于对照组(P<0.05)。患者血清5-HT含量与ISI评分呈负相关(r=-0.342,P<0.01);SOD含量与SCORAD评分呈负相关(r=-0.277,P<0.05),与ISI评分呈负相关(r=-0.311,P<0.01);MDA含量与SCORAD评分呈正相关(r=0.329,P<0.01)。疏肝健脾颗粒安全性良好,干预前后血常规及肝、肾功能未见异常改变。结论疏肝健脾颗粒对轻中度AD合并SDs患者疗效显著,其机制可能与抑制氧化应激反应、调节血清5-HT胺含量有关。

Bioactivity of five components of Chinese herbal formula Jiangzhi granules against hepatocellular steatosis

OBJECTIVE： This study aims to evaluate the bioactivity of five components of the traditional Chinese medicine complex prescription Jiangzhi granules against hepatocellular steatosis. METHODS： The five major components, including protopanaxadiol, tanshinone IIA, emodin, chlorogenic acid, and nuciferine, were extracted from Jiangzhi granules. Their cytotoxicity was assessed to determine the safe dose of each component for HepG2 cells. HepG2 cellular steatosis was induced using 1 mmol/L of free fatty acids （FFAs） for 24 h, and then treated with each component at high, intermediate, and low doses （500, 50, and 5 μmol/L）, respectively for another 24 h. The effects on HepG2 steatosis were observed directly under optical phase microscopy, or through oil red O staining and Nile red assays. In addition, the levels of reactive oxygen species （ROS） in the steatotic HepG2 cells with and without high-dose protopanaxadiol treatment were measured using fluorescent dye 2＇,7＇-dichlorodihydrofluorescein diacetate staining. RESULTS： No obvious cytotoxicity was observed in the HepG2 cells incubated with each of the five components at up to 500μmol/L. At 24 h after incubation with FFAs, the HepG2 cells swelled and many lipid droplets accumulated. The lipid content was attenuated after 24 h of incubation with protopanaxadiol, tanshinone IIA, and emodin at 500 or 50 μmol/L （P 〈 0.05）, especially with 500 μmol/L protopanaxadiol （P 〈 0.01）. In addition, the ROS level was elevated in steatotic cells, but decreased after intervention with 500μmol/L protopanaxadiol （P 〈 0.05）. CONCLUSION： Protopanaxadiol, tanshinone IIA, and emodin alleviate hepatocellular steatosis in a dose-dependent manner, and oxidative stress regulation may partially contribute to the effects of protopanaxadiol. ：

Biomechanical analysis of optic nerve injury treated by compound light granules and ciliary neurotrophic factor

In this study, rabbit models of optic nerve injury were reproduced by the clamp method. After modeling, rabbit models were given one injection of 50 ng recombinant human ciliary neurotrophic factor into the vitreous body and/or intragastric injection of 4 g/kg compound light granules containing Radix Angelicae Sinensis and Raidix Paeoniae Alba at 4 days after modeling, once per day for 30 consecutive days. After administration, the animals were sacrificed and the intraorbital optic nerve was harvested. Hematoxylin-eosin staining revealed that the injured optic nerve was thinner and optic nerve fibers were irregular. After treatment with recombinant human ciliary neurotrophic factor, the arrangement of optic nerve fibers was disordered but they were not markedly thinner. After treatment with compound light granules, the arrangement of optic nerve fibers was slightly disordered and their structure was intact. After combined treatment with recombinant human ciliary neurotrophic factor and compound light granules, the arrangement of optic nerve fibers was slightly disordered and the degree of injury was less than after either treatment alone. Results of tensile mechanical testing of the optic nerve showed that the tensile elastic limit strain, elastic limit stress, maximum stress and maximum strain of the injured optic nerve were significantly lower than the normal optic nerve. After treatment with recombinant human ciliary neurotrophic factor and/or compound light granules, the tensile elastic limit strain, elastic limit stress, maximum stress and maximum strain of the injured optic nerve were significantly increased, especially after the combined treatment. These experimental findings indicate that compound light granules and ciliary neurotrophic factor can alleviate optic nerve injury at the histological and biochemical levels, and the combined treatment is more effective than either treatment alone.

Application of moderate nitrogen levels alleviates yield loss and grain quality deterioration caused by post-silking heat stress in fresh waxy maize

High temperature(HT)during grain filling is one of the most important environmental factors limiting maize yield and grain quality.Nitrogen(N)fertilizer is essential for maintaining normal plant growth and defense against environmental stresses.The effects of three N rates and two temperature regimes on the grain yield and quality of fresh waxy maize were studied using the hybrids Suyunuo 5(SYN5)and Yunuo 7(YN7)as materials.N application rates were 1.5,4.5,and 7.5 g plant-1,representing low,moderate,and high N levels(LN,MN,and HN,respectively).Mean day/night temperatures during the grain filling of spring-and summer-sown plants were 27.6/21.0°C and 28.6/20.0°C for ambient temperature(AT)and 35/21.0°C and 35/20.0°C for HT,respectively.On average,HT reduced kernel number,weight,yield,and moisture content by 29.8%,17.9%,38.7%,and 3.3%,respectively.Kernel number,weight,yield,moisture,and starch contents were highest under MN among the three N rates under both temperature regimes.HT reduced grain starch content at all N levels.HT increased grain protein content,which gradually increased with N rate.Mean starch granule size under MN was larger(10.9μm)than that under LN and HN(both 10.4μm)at AT.However,the mean size of starch granules was higher under LN(11.7μm)and lower under MN(11.2μm)at HT.Iodine binding capacity(IBC)was lowest under MN and highest under HN among the three N levels under both temperature regimes.In general,IBC at all N rates was increased by HT.Peak viscosity(PV)was gradually reduced with increasing N rate at AT.In comparison with LN,PV was increased by MN and decreased by HN at HT.Retrogradation percentage gradually increased with N rate at AT,but was lowest under MN among the three N rates at HT.LN+AT and MN+HT produced grain with high pasting viscosity and low retrogradation tendency.MN application could alleviate the negative effects of HT on the grain yield and quality of fresh waxy maize.

Multi-scale analysis of subgrid stress and energy dissipation in turbulent channel flow

In present study, the subgrid scale （SGS） stress and dissipation for multiscale formulation of large eddy simulation are analyzed using the data of turbulent channel flow at Ret = 180 obtained by direct numerical simulation. It is found that the small scale SGS stress is much smaller than the large scale SGS stress for all the stress components. The dominant contributor to large scale SGS stress is the cross stress between small scale and subgrid scale motions, while the cross stress between large scale and subgrid scale motions make major contributions to small scale SGS stress. The energy transfer from resolved large scales to subgrid scales is mainly caused by SGS Reynolds stress, while that between resolved small scales and subgrid scales are mainly due to the cross stress. The multiscale formulation of SGS models are evaluated a priori, and it is found that the small- small model is superior to other variants in terms of SGS dissipation.

Experimental study on Tangshenjiangzhuo granules treatment of rats with early diabetic nephropathy

Objective:To study the effect of Tangshenjiangzhuo granules on serum cytokines as well as the oxidative stress and endoplasmic reticulum stress in kidney tissue of diabetic rats. Methods:SPF male Wistar rats were selected as experimental animals and randomly divided into control group, model group and intervention group. Model group received intragastric administration of saline after diabetes model was established, and the intervention group received intragastric administration of Tangshenjiangzhuo granules after diabetes model was established. 8 weeks after treatment, serum was collected to detect the levels of pro-inflammatory cytokines and adipocytokines, and kidney tissue was collected to detect the levels of oxidative stress molecules and endoplasmic reticulum stress molecules. Results:Serum interleukin-6 (IL-6), macrophage migration inhibitory factor (MIF), regulated upon activation normal T cell expressed secreted (RANTES), Chemerin and Nesfatin levels of model group were significantly higher than those of control group (P<0.05), and serum IL-6, MIF, RANTES, Chemerin and Nesfatin levels of intervention group were significantly lower than those of model group (P<0.05);reactive oxygen species (ROS), malondialdehyde (MDA), advanced oxidation protein products (AOPP), 8-hydroxy-2-deoxyguanosine (8-OHdG), glucose-regulated protein (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP) and ATF4 protein levels in kidney tissue of model group were significantly higher than those of control group (P<0.05), and ROS, MDA, AOPP, 8-OHdG, GRP78, CHOP and ATF4 levels in kidney tissue of intervention group were significantly lower than model group (P<0.05). Conclusions:Tangshenjiangzhuo granules can adjust the secretion of pro-inflammatory factors and adipocytokines, and inhibit oxidative stress and endoplasmic reticulum stress in kidney tissue of diabetic rat models.

RNA-binding proteins related to stress response and differentiation in protozoa

RNA-binding proteins(RBPs) are key regulators of gene expression. There are several distinct families of RBPs and they are involved in the cellular response to environmental changes, cell differentiation and cell death. The RBPs can differentially combine with RNA molecules and form ribonucleoprotein(RNP) complexes, defining the function and fate of RNA molecules in the cell. RBPs display diverse domains that allow them to be categorized into distinct families. They play important roles in the cellular response to physiological stress, in cell differentiation, and, it is believed, in the cellular localization of certain mRNAs. In several protozoa, a physiological stress(nutritional, temperature or pH) triggers differentiation to a distinct developmental stage. Most of the RBPs characterized in protozoa arise from trypanosomatids. In these protozoa gene expression regulation is mostly post-transcriptional, which suggests that some RBPs might display regulatory functions distinct from those described for other eukaryotes. mRNA stability can be altered as a response to stress. Transcripts are sequestered to RNA granules that ultimately modulate their availability to the translation machinery, storage or degradation, depending on the associated proteins. These aggregates of mRNPs containing mRNAs that are not being translated colocalize in cytoplasmic foci, and their numbers and size vary according to cell conditions such as oxidative stress, nutritional status and treatment with drugs that inhibit translation.

针刺联合息风化痰通络颗粒对急性脑梗死溶栓患者的影响及机制研究

目的:评估针刺联合息风化痰通络颗粒对急性脑梗死(ACI)后溶栓患者的疗效,探讨其作用机制。方法:纳入ACI溶栓患者178例,按随机数字表法分为观察组和对照组各89例,最终164例完成研究,观察组81例,对照组83例。对照组接受常规治疗,观察组在对照组的基础上加用针刺联合息风化痰通络颗粒治疗,两组均治疗2周。评估比较两组的临床疗效、美国国立卫生研究院卒中量表(NIHSS)评分、日常生活活动能力量表(ADL)评分、血清炎症因子水平、氧化应激因子水平以及脑血流动力学。结果:治疗后,两组ADL评分、白细胞介素-10(IL-10)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)水平均较治疗前升高(P<0.05),NIHSS评分、白细胞介素-1β(IL-1β)、超敏C反应蛋白(hs-CRP)、丙二醛(MDA)水平较治疗前降低(P<0.05),观察组NIHSS评分、IL-1β、IL-10、hs-CRP、MDA水平低于对照组(P<0.05),ADL评分、SOD、GSH-Px、CAT水平高于对照组(P<0.05)。两组脑血流动力学指标均较治疗前改善,且观察组改善情况优于对照组(P<0.05)。观察组治疗总有效率95.06%,对照组84.34%,两组疗效比较,差异有统计学意义(P<0.05)。结论:针刺联合息风化痰通络颗粒能够促进ACI溶栓患者脑血流动力学恢复,改善缺血再灌注损伤的神经功能,其作用机制可能与减轻炎症反应、抑制氧化应激反应有关。

小儿肺咳颗粒联合耳穴压豆治疗支气管哮喘患儿疗效观察

目的小儿肺咳颗粒联合耳穴压豆对慢性持续期支气管哮喘患儿氧化应激指标、肺功能和辅助性T淋巴细胞(T Helper Cells,Th1)/Th2平衡的影响。方法2020年4月—2023年4月该院儿科共收治了150例处于支气管哮喘慢性持续期的患儿。所有患儿通过随机数字表法被分配到对照组和试验组。对照组患儿接受吸入布地奈德混悬液的治疗,而试验组患儿在对照组的治疗基础上额外接受了小儿肺咳颗粒结合耳穴压豆的治疗方法。治疗4周后评价两组患者临床疗效、观察两组患者治疗前后氧化应激指标[超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(myeloperoxidase,MPO)和丙二醛(malondialdehyde,MDA)]水平,肺功能指标[肺活量(forced vital capacity,FVC)、1秒用力呯吸量(forced expiratory volume in one second,FEV1)和最大呼气峰流速(peak expiratory flow,PEF)]以及Th1/Th2平衡相关因子[白细胞介素-12(interleukin-12,IL-12)、干扰素-γ(interferon-γ,IFN-γ)、白细胞介素-4(interleukin-4,IL-4)、白细胞介素-13(interleukin-13,IL-13)]指标的变化。结果两组患儿一般资料比较,差异无统计学意义(P>0.05)。治疗结束后,试验组总有效率为84.84%(56/66),显著高于对照组75.71%(53/70),差异有统计学意义(P<0.05)。治疗后,两组受试者的血清SOD水平有所提升,而MPO和MDA水平则明显下降(P<0.05)。此外,治疗后,两组的肺功能指标均有所改善(P<0.05),且试验组的肺功能各项指标均优于对照组(P<0.05)。治疗后,两组患儿血清IL-12和IFN-γ较治疗前升高,IL-4和IL-13水平较治疗前降低(P<0.05)。治疗后,相较于对照组,试验组患儿的血清IL-12和IFN-γ含量显著增加,同时IL-4和IL-13的含量则显著减少(P<0.05)。结论对于儿童支气管哮喘慢性持续期,小儿肺咳颗粒结合耳穴压豆疗法不仅能有效提升患儿的肺功能和降低氧化应激水平,还能调节Th1/Th2免疫平衡,且具有较高的安全性。

疏肝利胆颗粒治疗原发性胆汁性胆管炎72例的疗效观察

目的:探讨疏肝利胆颗粒治疗原发性胆汁性胆管炎(PBC)胆腑郁热证患者效果。方法:纳入兰州大学第一医院普外科就诊的72例胆腑郁热证PBC患者,根据数字奇偶法分为对照组和治疗组,每组36例。对照组服用熊去氧胆酸胶囊治疗,治疗组服用熊去氧胆酸胶囊+疏肝利胆颗粒治疗。比较两组临床症状、微炎症指标[白细胞介素6(IL-6)、白细胞介素17(IL-17)、肿瘤坏死因子-α(TNF-α)]、肝功能[谷草转氨酶(AST)、总胆红素(TBIL)、谷丙转氨酶(ALT)]、氧化应激指标[丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)]、肝纤维化[血清透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原、Ⅳ型胶原]、免疫球蛋白M(IgM)变化及疗效情况。结果:两组治疗后身热不退、身目发黄、胁肋胀痛、大便秘结、呕吐呃逆、口苦咽干中医证候积分对比,治疗组均低于对照组(P<0.05)。两组治疗后IL-17、IL-6、TNF-α水平均下降(P<0.05),治疗组比对照组更低(P<0.05)。两组TBIL、AST、ALT水平治疗后均下降(P<0.05),治疗组比对照组更低(P<0.05)。两组治疗后MDA水平均降低(P<0.05),治疗组更低(P<0.05),两组治疗后GSH-Px、SOD水平均升高(P<0.05),治疗组更高(P<0.05)。两组治疗后肝纤维化、免疫指标HA、LN、III型前胶原、IV型胶原、IgM水平均下降(P<0.05),治疗组比对照组更低(P<0.05)。治疗后治疗组总有效率显著高于对照组(91.67%vs 72.22%)。结论:疏肝利胆颗粒治疗胆腑郁热证PBC临床疗效显著,能有效促进临床症状及肝功能改善,减轻机体炎症反应及氧化应激反应,改善机体免疫功能、肝纤维化程度,具有较高的临床推广价值。

基于网络药理学和实验验证探究生血通便颗粒治疗慢传输型便秘的作用机制

目的采用网络药理学预测生血通便颗粒治疗慢传输型便秘(slow transit constipation,STC)的潜在作用机制,并进行动物实验验证。方法利用TCMSP数据库和BATMAN-TCM数据库筛选生血通便颗粒的活性成分及作用靶点,通过GeneCards数据库和OMIM数据库筛选STC疾病相关靶点,构建“活性成分-疾病靶点”网络,使用R语言对交集基因进行GO及KEGG富集分析。大鼠采用盐酸洛哌丁胺混悬液[3 mg/(kg·d)]灌胃建立STC模型。将大鼠分为正常组(等体积蒸馏水)、模型组(等体积蒸馏水)、莫沙比利组[1.35 mg/(kg·d)]、生血通便颗粒低剂量组[1.44 g/(kg·d)]、生血通便颗粒高剂量组[2.88 g/(kg·d)],每组8只,每日灌胃1次,连续治疗14 d。记录治疗后肠道推进率;HE染色观察结肠组织病理学改变;免疫组织化学法检测结肠组织酪氨酸激酶受体(tyrosine kinase receptor,c-Kit)表达;TUNEL染色检测结肠组织Cajal间质细胞(interstitial cells of Cajal,ICC)凋亡情况;Western blot检测结肠组织葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)、C/EBP同源蛋白(C/EBP homologous protein,CHOP)、B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、活化型胱天蛋白酶-3(cleaved cysteine aspartic acid specific protease-3,cleaved Caspase-3)蛋白表达水平。结果筛选出生血通便颗粒活性成分74个,对应的作用靶点为492个,CASP3(Caspase-3)、Bcl-2、Bax凋亡基因是STC的核心靶点之一,其相关的作用通路为内质网应激信号通路。动物实验结果表明,与正常组比较,模型组大鼠肠道推进率降低(P<0.01);结肠黏膜萎缩、变薄,黏膜上皮层有缺失,固有层腺体缺损破坏明显,可见炎性改变;结肠组织中c-Kit表达降低(P<0.01);凋亡细胞绿色荧光增加、c-Kit红色荧光减少,ICC凋亡指数升高(P<0.01);结肠组织GRP78、CHOP、Bax、cleaved Caspase-3蛋白表达水平升高(P<0.01),Bcl-2蛋白表达水平降低(P<0.01)。与模型组相比,莫沙必利组及生血通便颗粒低、高剂量组大鼠肠道推进率提高(P<0.01);结肠黏膜结构较完整,炎性情况有所恢复,腺体排列较整齐;结肠组织中c-Kit表达升高(P<0.01);凋亡细胞绿色荧光减少、c-Kit红色荧光增加,ICC凋亡指数降低(P<0.01);结肠组织GRP78、CHOP、Bax、cleaved Caspase-3蛋白表达水平降低(P<0.01),Bcl-2蛋白表达水平升高(P<0.01)。与莫沙必利组相比,生血通便颗粒高剂量组肠道推进率提高(P<0.05)、c-Kit表达升高(P<0.05)、ICC凋亡指数降低(P<0.05);生血通便颗粒低、高剂量组结肠组织中GRP78、CHOP、Bax、cleaved Caspase-3蛋白表达水平降低(P<0.01),生血通便颗粒高剂量组Bcl-2蛋白表达水平升高(P<0.01)。结论生血通便颗粒可通过抑制内质网应激GRP78/CHOP信号通路,调控Bax、Bcl-2、cleaved Caspase-3凋亡蛋白表达,从而抑制ICC凋亡以达到治疗STC的作用。

坤宁颗粒对痛经模型大鼠的抗炎镇痛作用研究

[目的]研究坤宁颗粒对痛经模型大鼠COX-2/PGF2α通路、内分泌激素、炎症因子和氧化应激水平的影响,探讨其保护作用及机制。[方法]取SD雌性大鼠,随机分为6组,正常对照组、模型组、益母草颗粒(3.5 g/kg)阳性对照组、坤宁颗粒低(1.4 g/kg)、中(2.8 g/kg)、高(5.6 g/kg)剂量组,除正常对照组外,其余组建立痛经模型,第1、10天灌胃戊酸雌二醇每只0.2 mg,第2~9天每只0.1 mg,从第5天各给药组灌胃给药,正常对照组和模型组灌胃蒸馏水,每日1次,第10天正常对照组大鼠腹腔注射生理盐水,其余各组大鼠腹腔注射缩宫素每只10 U。观察各组大鼠扭体反应次数;采用酶联免疫吸附实验(ELISA)检测大鼠血清前列腺素F2α(PGF2α)、前列腺素E_(2)(PGE_(2))、β-内啡肽(β-EP)、雌二醇(E_(2))、孕酮(PROG)、白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;比色法检测大鼠血清超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)水平;蛋白免疫印迹法(Western Blot)检测大鼠子宫组织环氧合酶-2(COX-2)、前列腺素E_(2)受体(PTGER2)、雌激素受体α(ERα)蛋白表达;苏木精-伊红(HE)染色观察子宫组织病理形态。[结果]与正常对照组相比,模型组大鼠扭体反应次数显著增加,大鼠血清中PGF2α、E_(2)、IL-1β、IL-6、TNF-α、MDA水平和子宫组织COX-2、ERα蛋白表达显著升高,而血清PGE_(2)、β-EP、PROG、SOD、GSH-Px水平显著降低,子宫PTGER2蛋白表达显著降低,子宫组织可见明显充血、水肿。与模型组相比,坤宁颗粒可以显著降低大鼠扭体反应次数和血清中PGF2α、E_(2)、IL-1β、IL-6、TNF-α、MDA水平,升高PGE_(2)、β-EP、PROG、SOD、GSH-Px水平,显著抑制子宫组织COX-2、ERα蛋白表达,升高PTGER2蛋白表达,组织病理学明显改善。[结论]坤宁颗粒对痛经模型大鼠具有显著的改善作用,其机制可能与调节COX-2/PGF2α通路和PGE_(2)、β-EP、E_(2)、PROG等内分泌激素、降低炎症因子水平、改善氧化应激水平有关。