Soybean(Glycine max)rhizosphere organic phosphorus recycling relies on acid phosphatase activity and specific phosphorusmineralizing-related bacteria in phosphate deficient acidic soils

Bacteria play critical roles in regulating soil phosphorus(P) cycling. The effects of interactions between crops and soil P-availability on bacterial communities and the feedback regulation of soil P cycling by the bacterial community modifications are poorly understood. Here, six soybean(Glycine max) genotypes with differences in P efficiency were cultivated in acidic soils with long-term sufficient or deficient P-fertilizer treatments. The acid phosphatase(AcP) activities, organic-P concentrations and associated bacterial community compositions were determined in bulk and rhizosphere soils. The results showed that both soybean plant P content and the soil AcP activity were negatively correlated with soil organic-P concentration in P-deficient acidic soils. Soil P-availability affected the ɑ-diversity of bacteria in both bulk and rhizosphere soils. However, soybean had a stronger effect on the bacterial community composition, as reflected by the similar biomarker bacteria in the rhizosphere soils in both P-treatments. The relative abundance of biomarker bacteria Proteobacteria was strongly correlated with soil organic-P concentration and AcP activity in low-P treatments. Further high-throughput sequencing of the phoC gene revealed an obvious shift in Proteobacteria groups between bulk soils and rhizosphere soils, which was emphasized by the higher relative abundances of Cupriavidus and Klebsiella, and lower relative abundance of Xanthomonas in rhizosphere soils. Among them, Cupriavidus was the dominant phoC bacterial genus, and it was negatively correlated with the soil organic-P concentration. These findings suggest that soybean growth relies on organic-P mineralization in P-deficient acidic soils, which might be partially achieved by recruiting specific phoCharboring bacteria, such as Cupriavidus.

Effects of heat shock on change of HSC70/HSP68,acid and alkaline phosphatases before and after rat partial hepatectomy

INTRODUCTIONOnly the liver has the great capability ofregeneration in mammal.Few hepatocytes are inthe phase of division in the normal liver of an adultmammal (including human beings),but theremaining hepatocytes can be induced to proliferatequickly by partial hepatectomy (PH),and,to somedegree,they stop dividing and re-differentiate intocells functioning as hepatocytes.This shows

Molecular Characterization and Functional Analysis of a New Acid Phosphatase Gene(Ha-acp1) from Heterodera avenae

For sedentary endo-parasitic nematodes, parasitism genes encoding secretory protein expressed in the subventral glands cells always play an important role during the early parasitic process. A new acid phosphatase gene （Ha-acp1） expressed in the subventral glands of the cereal cyst nematode （Heterodera avenae） was cloned and the characteristics of the gene were analyzed. Results showed that the gene had a putative signal peptide for secretion and in situ hybridization showed that the transcripts of Ha-acp1 accumulated speciifcally in the subventral gland cells of H. avenae. Southern blot analysis suggested that Ha-acp1 belonged to a multigene family. RT-PCR analysis indicated that this transcription was strong at the pre-parasitic juveniles. Knocking down Ha-acp1 using RNA interference technology could reduce nematode infectivity by 50%, and suppress the development of cyst. Results indicated that Ha-acp1 could play an important role in destroying the defense system of host plants.

Pachymic acid exerts antitumor activities by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B

Objective:To determine the inhibitory effects of pachymic acid on lung adenocarcinoma(LUAD)cells and elucidate its underlying mechanism.Methods:CCK-8,wound healing,Transwell,Western blot,tube formation,and immunofluorescence assays were carried out to measure the effects of various concentrations of pachymic acid on LUAD cell proliferation,metastasis,angiogenesis as well as autophagy.Subsequently,molecular docking technology was used to detect the potential targeted binding association between pachymic acid and protein tyrosine phosphatase 1B(PTP1B).Moreover,PTP1B was overexpressed in A549 cells to detect the specific mechanisms of pachymic acid.Results:Pachymic acid suppressed LUAD cell viability,metastasis as well as angiogenesis while inducing cell autophagy.It also targeted PTP1B and lowered PTP1B expression.However,PTP1B overexpression reversed the effects of pachymic acid on metastasis,angiogenesis,and autophagy as well as the expression of Wnt3a andβ-catenin in LUAD cells.Conclusions:Pachymic acid inhibits metastasis and angiogenesis,and promotes autophagy in LUAD cells by modulating the Wnt/β-catenin signaling pathway via targeting PTP1B.

Mammalian-like Purple Acid Phosphatases in Plants

Introduction Purple acid phosphatases （PAPs） comprise of a family of binuclear metal-containing hydrolases, some members of which have been isolated and characterized from animal, plant and fungal sources . PAPs not only catalyze the hydrolyses of a wide range of phosphate esters and anhydrides under acidic reaction conditions, but also catalyze the generation of hydroxyl radicals in a Fenton-like reaction, by virtue of the presence of a redox-active binuclear metal center. Inmammals,

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase （APase） exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine （Pinus tabulaeformis）, under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, differed under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the different ions played different roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.

Effect of acid phosphatase produced by Trichoderma asperellum Q1 on growth of Arabidopsis under salt stress

Salt stress is a major environmental factor that inhibits crop growth.Trichoderma spp.are the most efficient biocontrol fungi and some of the strains can stimulate plant growth.Phosphate solubilization is known as one of the main mechanisms in promoting plant growth,but the underlying mechanisms of phosphate solubilization in the salinity still need to be explored.The Trichoderma asperellum Q1 isolated and identified in our lab is a beneficial rhizosphere biocontrol fungus with a high phosphate solubilization activity.It could produce acid and alkaline phosphatases when using insoluble organic phosphorus as the sole phosphorus source,the salt stress increased the phosphorus-solubilization ability of the strain and the activities of the two enzymes.Furthermore,an acid phosphatase was purified from the fermentation broth by ammonium sulphate precipitation,ion-exchange,and gel filtration chromatography.Its molecular weight was 55 k Da as determined by SDS-PAGE.The purified acid phosphatase was used to investigate growth performance of Arabidopsis thaliana by plate assay and the result showed that it contributed to Arabidopsis growth by transforming organic phosphate into a soluble inorganic form under salt stress.To our knowledge,this is the first report on acid phosphatase purification from T.asperellum and its function in regulation of plant growth under salt stress.

Effects of Lanthanum and Cerium on Acid Phosphatase Activities in Two Soils

To evaluate the security of using thulium, comparision between effects of La and those of Ce on acidic phosphatase activities in red soil and yellow soil in Zhejiang district was studied under conditions of ambient temperature and humidity. Results show that the acid phosphatase from different soil respondes to La and Ce differently. The activity of acid phosphatase in soil 1 declines with the increase of the concentration of La and Ce. The maximum inhibitory ratio of La and Ce reaches 69.8% and 71.0% respectively. But La and Ce have stimulative effect on the activity of acid phosphatase in soil 2. Under the effect of same concentration of the thulium, the acid phosphatase in two soils increases with the extending of culture time.

Detection of Ca^(2+)-dependent acid phosphatase activity identifies neuronal integrity in damaged rat central nervous system after application of bacterial melanin

The study aims to confirm the neuroregenerative effects of bacterial melanin （BM） on central nervous system injury using a special staining method based on the detection of Ca^2＋-dependent acid phosphatase activity. Twenty-four rats were randomly assigned to undergo either unilateral destruction of sensorimotor cortex （group I; n = 12） or unilateral rubrospinal tract transection at the cervical level （C3-4） （group II; n = 12）. In each group, six rats were randomly selected after surgery to undergo intramuscular injection of BM solution （BM subgroup） and the remaining six rats were intramuscularly in）ected with saline （saline subgroup）. Neurological testing confirmed that BM accelerated the recovery of motor function in rats from both BM and saline subgroups. Two months after surgery, Ca^2＋-dependent acid phosphatase activity detection in combination with Chilingarian＇s calcium adenoside triphosphate method revealed that BM stimulated the sprouting of fibers and dilated the capillaries in the brain and spinal cord. These results suggest that BM can promote the recovery of motor function of rats with central nervous system injury; and detection of Ca^2＋-dependent acid phosphatase activity is a fast and easy method used to study the regeneration-promoting effects of BM on the injured central nervous system.

Adsorption of Acid Phosphatase on Minerals and Soil Colloids in Presence of Citrate and Phosphate

The aim of this work was to study the influence of phosphate and citrate,which are common inorganic and organic anions in soils,on the adsorption of acid phosphatase by kaolin,goethite and the colloids separated from yellow-brown soil(YBS)and latosol(LS)in central-south China.The YBS colloid has the major clay mincral composition of 1.4 nm mincral,illite and kaolinite while the LS colloid mainly contains kaolinite and oxides.The adsorption isotherm of acid phosphatase on the examined soil colloids and minerals fitted to the Langmuir model.The amount of enzyme adsorbed in the absence of ligands was in the order of YBS colloid>LS colloid>kaolin≈goethite.In the presence of phosphate or citrate,the amounts of the enzyme adsorbed followed the sequence YBS colloid>kaolin>LS colloid>goethite.The presence of ligands also decreased the binding energy between the enzyme and soil colloids or minerals.With the increase of ligand concentration from 10mmol L^-1 to 400 m mol L^-1,different behaviors for the adsorption of cnzyme were found in the colloid and mineral systems studied.A sharp decrease in enzyme adsorption was observed on goethite while gradual decreases of enzyme adsorption were recorded in the two soil colloid systems.However,no any decrease was found for the amount of enzyme adsorbed on kaolin at higher ligand concentrations.When phosphate or citrate was introduced to the system before the addition of enzyme,the ligands usually enhanced the adsorption of enzyme.The results obtained in this study suggested the important role of kaolinite mineral in the adsorption of enzyme molecules in acidic soils in the presence of various ligands.

青稞酸性磷酸酶基因HvnACP2克隆和亚细胞定位研究

为了探索青稞酸性磷酸酶基因 HvnACP2的基因和蛋白结构特点,为青稞磷吸收利用机制研究提供基础。以青稞‘肚里黄’叶片为材料,根据植物基因组数据库Gramene中的大麦 HvACP2基因和启动子区域序列设计引物,通过PCR获得青稞 HvnACP2基因和启动子区域序列。采用生物信息学软件对 HvnACP2基因启动子区域元件以及蛋白理化性质、跨膜结构、磷酸化位点、信号肽、二级、三级结构进行分析。结果表明: HvnACP2有2个外显子和1个内含子,启动子区域有与分生组织、光响应、厌氧、茉莉酸和脱落酸相关的顺式作用元件。HvnACP2蛋白由454个氨基酸组成,分子质量为51 285.15 u,总原子数为7 058,亲水系数为-0.451,理论等电点为6.17,不稳定指数34.54,脂溶性指数为67.05。具有12个磷酸化位点和信号肽,不具有跨膜结构。 HvnACP2中α螺旋、延长链、β转角、无规则卷曲所占比例分别为18.72%、53.96%、23.57%、3.74%。 HvnACP2和其他物种的同源蛋白都有紫色酸性磷酸酶N末端结构域和金属酸性磷酸酶结构域,将 HvnACP2与其他植物的氨基酸序列进行对比并构建进化树,发现青稞 HvnACP2与小麦TaACP2亲缘关系最近。亚细胞定位结果显示 HvnACP2定位在细胞膜上。

Effects of Hg and Cu on the activities of soil acid phosphatase

Comparative study on the activity and kinectic properties of acid phosphatase (ACPase) of three soils amended with Hg and Cu at constant temperature and humidity was carried out. The results indicated that the inhibition on ACPase of the three sample soils by Hg and Cu varied with the content of soil organic matter and pH, where, Soil 1 was the most seriously contami- nated due to its lowest content of organic matter and the lowest pH among three samples, Soil 2 took the second place, and Soil 3 was the least contaminated. Except Soil 3, the activity of soil ACPase tended to increase along with the contact time under the same type and the same concentration of heavy metal. In particular the Vmax values of ACPase in all three samples decreased with increasing Hg and Cu concentration, whereas the Km values were affected weakly. According to the change of Vmax and Km values, Cu and Hg had the same inhibition effect on soil ACPase. Both of them may be a type of compound of non-competitive and anti-competitive inhibition. Statistic analyses indicated that activities of soil ACPase and Vmax values could serve as bioindicator to partially denote the heavy metal Hg and Cu contamination degree.

Effect of noise exposure(85 dB) on testicular adrenocortical steroidogenic key enzymes, acid and alkaline phosphatase activities of sex organs in mature albino rats

Changes in the activities of Δ 5\|3β\|hydroysteroid dehydrogenase(HSD) in testis and adrenal gland, 17β\|hydroxysteroid dehydrogenase in testis, acid and alkaline phosphatase in testis, prostate and seminal vesicle were observed in noise exposed mature rats at the intensity of 85 dB for 8 h/day for 45 days. The results indicated that noise exposed group showed a significant diminution in the activities of androgenic key enzymes Δ 5\|3β and 17β\|HSD, acid phosphatase in testis, prostate and seminal vesicle. There was a significant elevation in the activities of adrenal Δ 5\|3β\|HSD, alkaline phosphatase in testis and other accessory sex organ in noise exposed group. Gonadosomatic, prostatosomatic and seminal vesiculo\|somatic indexes were decreased significantly in noise exposed group. Therefore, it is evident that noise exposure at 85dB exerts a deleterious effect on testicular and adrenocortical activities.

血清MIP-1α、TRACP-5b和骨硬化蛋白水平在多发性骨髓瘤病情和预后中的临床价值

目的探讨血清巨噬细胞炎症蛋白-1α(MIP-1α)、抗酒石酸酸性磷酸酶5b(TRACP-5b)和骨硬化蛋白水平在多发性骨髓瘤病情和预后中的临床价值。方法选取2019年1月至2020年12月在该院就诊的125例多发性骨髓瘤患者作为观察组;另选取同期在该院体检的75例健康体检者作为对照组。比较观察组患者治疗前后血清MIP-1α、TRACP-5b和骨硬化蛋白水平,以及观察组和对照组血清MIP-1α、TRACP-5b和骨硬化蛋白水平;比较不同肿瘤分期、不同骨质破坏程度分级多发性骨髓瘤患者血清MIP-1α、TRACP-5b和骨硬化蛋白水平,分析多发性骨髓瘤患者1年内发生死亡的单因素和多因素;比较血清MIP-1α、TRACP-5b和骨硬化蛋白单项及3项指标联合检测对多发性骨髓瘤患者1年内发生死亡的预测效能。结果观察组治疗前后血清MIP-1α、TRACP-5b和骨硬化蛋白水平均明显高于对照组,差异均有统计学意义(P<0.05);观察组患者治疗后血清MIP-1α、TRACP-5b和骨硬化蛋白水平较治疗前均明显降低,差异均有统计学意义(P<0.05)。多发性骨髓瘤患者血清MIP-1α、TRACP-5b和骨硬化蛋白水平随着多发性骨髓瘤肿瘤分期和骨质破坏程度分级升高而升高,差异均有统计学意义(P<0.05)。死亡组患者肿瘤分期、骨质破坏程度分级、MIP-1α水平、TRACP-5b水平和骨硬化蛋白水平与存活组比较,差异均有统计学意义(P<0.05)。多因素分析结果显示,肿瘤分期、骨质破坏程度分级、MIP-1α水平、TRACP-5b水平和骨硬化蛋白水平是多发性骨髓瘤患者1年内发生死亡的影响因素(P<0.05)。血清MIP-1α、TRACP-5b和骨硬化蛋白水平对多发性骨髓瘤患者1年内发生死亡具有较高的预测效能,3项指标联合检测的灵敏度为90.0%,特异度为87.1%,受试者工作特征曲线下面积(AUC)为0.934,明显高于MIP-1α(Z=3.068,P=0.002)、TRACP-5b(Z=3.622,P<0.001)和骨硬化蛋白(Z=2.576,P=0.010)单独检测,而3项指标之间的AUC比较,差异无统计学意义(P>0.05)。结论MIP-1α、TRACP-5b和骨硬化蛋白参与了多发性骨髓瘤疾病的发生和发展,对预测其1年内发生死亡具有较高的价值。

血清MIP-1β、TRACP-5b和FGF-23检测对绝经后骨质疏松患者发生骨折的预测价值

目的探讨血清巨噬细胞炎症蛋白-1β(MIP-1β)、抗酒石酸酸性磷酸酶5b(TRACP-5b)和成纤维细胞生长因子23(FGF-23)检测对绝经后骨质疏松患者发生骨折的预测价值。方法选择2019年1月至2021年12月该院收治的绝经后骨质疏松患者121例纳入骨质疏松组,根据患者是否发生骨折分为骨折组(54例)和非骨折组(67例)。选择同期在该院诊断为骨量下降和骨量正常的绝经后女性分别纳入骨量下降组(82例)和对照组(65例)。比较各组血清MIP-1β、TRACP-5b和FGF-23水平,对骨质疏松患者发生骨折的影响因素进行单因素和多因素分析,评价血清MIP-1β、TRACP-5b和FGF-23检测对绝经后骨质疏松患者发生骨折的预测价值,以及分析血清MIP-1β、TRACP-5b和FGF-23水平与骨折严重程度的关系。结果骨质疏松组血清MIP-1β、TRACP-5b和FGF-23水平明显高于骨量下降组和对照组(P<0.01),骨量下降组血清MIP-1β、TRACP-5b和FGF-23水平明显高于对照组(P<0.01)。骨折组的血清MIP-1β、TRACP-5b、FGF-23水平明显高于非骨折组(P<0.01)。多因素分析发现,血清MIP-1β>719.72 pg/mL、TRACP-5b>6.97 U/L和FGF-23>247.32 pg/mL是绝经后骨质疏松患者发生骨折的危险因素(P<0.01)。血清MIP-1β、TRACP-5b和FGF-23水平在预测骨质疏松患者发生骨折方面具有较高的效能(P<0.01),3项指标联合检测灵敏度为90.7%,特异度为89.4%,曲线下面积(AUC)为0.946,AUC明显高于MIP-1β(Z=3.412,P<0.01)、TRACP-5b(Z=3.811,P<0.01)和FGF-23(Z=3.983,P<0.01)单项指标检测,而单项指标之间的AUC比较差异无统计学意义(P>0.05)。结论血清MIP-1β、TRACP-5b和FGF-23参与了绝经后骨质疏松的发生、发展过程,其水平检测对预测绝经后骨质疏松患者发生骨折具有较高的价值。

Effect of Salinity on Germination,Seedling Growth and Acid Phosphatase Activity in Lettuce

The impact of salt stress (NaCl 100 mM) on two lettuce varieties Romaine and Vista was conducted at germination and early seedling stages. The seeds of lettuce varieties were provided by the Seed Laboratory of Tunisian Ministry of Agriculture. The seeds were germinated in Petri dishes with double filter paper in distilled water (control) or NaCl solution (100 mM) for 5 days. The result showed that salinity significantly affected percentage and rate of germination in Vista variety but 100% of germination was found in Romaine. Length and fresh weight of root and shoot were reduced significantly with salt treatment in two lettuce varieties. Regarding biochemical analysis, acid phosphatase activity in root increased in Romaine and decreased in Vista. In shoot, this activity showed no difference with the control in the two varieties. However in cotyledons, and during 24 hours after germination, salinity decreased acid phosphatase activity in both varieties whereas in the later hours (48 - 96 h) this activity reached the value of the control in Romaine and Vista.

乳腺癌患者血清PCSK9、Tracp5b及MUC1表达水平及其与肿瘤病理特征的相关性分析

目的分析乳腺癌患者血清前蛋白转化酶枯草溶菌素9(PCSK9)、抗酒石酸酸性磷酸酶5b(Tracp5b)及黏蛋白1(MUC1)的表达水平及其与肿瘤病理特征的相关性。方法选择该院2019年1月至2022年1月收治的106例乳腺癌患者作为观察组,另选择同期在该院体检的106例健康体检女性作为对照组。检测两组血清PCSK9、Tracp5b及MUC1表达水平,并分析不同肿瘤病理特征的乳腺癌患者血清PCSK9、Tracp5b及MUC1表达水平。采用Spearman相关分析血清PCSK9、Tracp5b及MUC1与肿瘤最大径、病理分期、分化程度的关系。绘制受试者工作特征(ROC)曲线分析血清PCSK9、Tracp5b及MUC1对腋窝淋巴结转移的预测效能。结果观察组血清PCSK9、Tracp5b及MUC1表达水平均高于对照组,差异均有统计学意义(P<0.05)。肿瘤最大径>2 cm患者血清PCSK9、Tracp5b及MUC1表达水平均高于肿瘤最大径≤2 cm患者,差异均有统计学意义(P<0.05);Ⅰ~Ⅲ期病理分期及低、中、高分化患者血清PCSK9、Tracp5b及MUC1表达水平比较,差异均有统计学意义(P<0.05)。Spearman相关分析结果显示,乳腺癌患者血清PCSK9、Tracp5b及MUC1表达水平均与肿瘤最大径、病理分期呈正相关(P<0.05),与分化程度呈负相关(P<0.05)。106例乳腺癌患者中,发生腋窝淋巴结转移20例为淋巴结转移组,剩余86例为淋巴结未转移组,淋巴结转移组血清PCSK9、Tracp5b及MUC1表达水平均高于非淋巴结转移组,差异有统计学意义(P<0.05)。ROC曲线分析结果显示,乳腺癌患者血清PCSK9、Tracp5b与MUC1联合预测腋窝淋巴结转移的曲线下面积(AUC)为0.896,明显大于PCSK9、Tracp5b和MUC1单项预测(AUC为0.668、0.644、0.697),差异均有统计学意义(Z=2.365、2.613、2.874,P<0.05)。结论乳腺癌患者血清PCSK9、Tracp5b及MUC1表达水平明显上调,均与肿瘤最大径、病理分期、分化程度密切相关,联合预测腋窝淋巴结转移的效能较好,值得临床予以重视。

Acid-Phosphorus Activity of Wheat Varieties Rhizobacteria of Uzbekistan

In this work, local strains of phosphate-solubilizing microorganisms were isolated and identified from the wheat rhizosphere and exogenous acid phosphatase enzymes of locally active phosphate- and potassium-mobilizing rhizobacteria belonging to the genera Escherichia, Rahnella, Bacillus, Enterobacter, Pseudomonas, and Pantoea were studied. The efficiency of the physiological properties of rhizobacteria is determined by the production of soluble phosphorus, and the amount of phosphorus depends on the activity and biomass of bacteria that secrete phosphorus. This is done by phosphate solubilizing bacteria, and the habitat ecosystem is enriched with beneficial micronutrients. In these studies, active rhizobacteria activity of acid phosphatase in nutrient liquid was studied at different temperatures. Optimum pH activity index and temperature variability of enzymes were determined. It should be noted that in the most active phosphate-solubilizing strains the maximum enzymatic activity was observed in the culture fluid of R. aquatilis strain 17, which produced 1.086 μmol p-nitrophenol μmol/min/ml. P. agglomerans 22, P. agglomerans 20 and Ps. kilonensis 32 cultures phosphatase activity was 0.143 - 0.680 p-nitrophenol μmol/min/ml. It should be noted that the phosphatase activity of bacteria belonging to the same genus and species was very different from each other. That is, the enzyme activity of Rahnella aquatilis strain 17 was 9 times higher than the enzyme activity of Rahnella aquatilis strain 9. The pH optimum of sour phosphatase enzymes in Rahnella aquatilis strain 16 was 6.0. The optimum temperature of acid phosphatase activity was 45˚C and 50˚C. The reason for this may be that the strains were isolated in different soil and climate conditions. When the acid phosphatase activity of R. aquatilis 3, 9, E. cloacae 8 and P. agglomerans 22 cultures was determined at a temperature of 45˚C, it was observed that the enzyme activity increased by 2 - 4 times. Es. hermannii 1, Ps. kilonensis 26 and B. simplex 28 bacteria acid phosphatase activity was not significantly affected by temperature rise.

非小细胞肺癌组织中lncRNA GAS5、LHPP表达与上皮间质化相关性及临床意义

目的探讨非小细胞肺癌(NSCLC)患者癌组织中长链非编码核糖核酸生长抑制特异性基因5(lncRNA GAS5)、磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶(LHPP)表达与上皮间质化(EMT)相关性及临床意义。方法收集2018年6月至2020年1月在河北北方学院附属第一医院行手术切除的NSCLC 90例患者的癌组织及癌旁组织,采用实时荧光定量聚合酶链反应检测lncRNA GAS5、LHPP和EMT相关蛋白[E-钙黏蛋白(E-Cad)、N-钙黏蛋白(N-Cad)和波形蛋白(VIM)]表达。分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与临床病理特征的关系,并通过Pearson相关性分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与EMT相关蛋白表达的相关性。采用Kaplan-Meier法绘制不同lncRNA GAS5、LHPP mRNA表达的NSCLC患者生存曲线,多因素Cox回归分析NSCLC患者预后的影响因素。结果NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA、E-Cad mRNA表达低于癌旁组织,N-Cad mRNA、VIM mRNA表达高于癌旁组织,差异有统计学意义(P<0.05)。Pearson相关性分析显示,NSCLC患者癌组织中lncRNA GAS5与E-Cad mRNA表达呈正相关(r=0.724,P<0.001),与N-Cad mRNA、VIM mRNA表达呈负相关(r=-0.699、-0.689,P<0.001);lncRNA GAS5与LHPP mRNA表达呈正相关(r=0.651,P<0.001)。不同分化程度、肿瘤TNM分期、淋巴结转移的NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA表达比较差异有统计学意义(P<0.05)。Kaplan-Meier生存曲线分析显示,lncRNA GAS5高表达组的3年总生存率[68.18%(30/44)]高于lncRNA GAS5低表达组的3年总生存率[36.96%(17/46)];LHPP mRNA高表达组的3年总生存率[67.39%(31/46)]高于LHPP mRNA低表达组的3年总生存率[36.36%(16/44)],差异有统计学意义(χ^(2)=10.274、10.322,P<0.05)。低分化、肿瘤TNM分期Ⅲ期、有淋巴结转移为NSCLC患者死亡的独立危险因素,lncRNA GAS5≥1.32、LHPP mRNA≥1.12为独立保护因素(P<0.05)。结论NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA低表达,与EMT相关蛋白表达、分化程度、肿瘤TNM分期、淋巴结转移和预后有关,可能成为NSCLC诊治的新靶点。

新型酸性磷酸酶“Turn-on”型荧光底物

酸性磷酸酶(ACP)是前列腺癌、戈谢病、肾病等疾病的重要生物标记物,因此开发灵敏、高选择性的ACP检测方法具有重要的意义。目前,已发展出多种ACP检测方法,包括免疫法、光谱法、色谱法、电化学法等;其中,荧光检测方法因灵敏度高、选择性好、快速、准确等优点备受关注。通过席夫碱反应,利用2-氨基苯硼酸(2-APBA)二聚体和磷酸吡哆醛(PLP)合成了一种新型的ACP“Turn-on”型荧光底物APBA-PLP(ABPL)。反应后,PLP的—CHO基团和2-APBA二聚体的—NH 2基团的特征峰消失,同时ABPL的C N基团的特征峰出现;此外,^(1)H和^(1)H—^(1)H COSY核磁波谱的信号峰与ABPL的结构相对应。以上表征结果证实了ABPL的成功合成。进一步,将ABPL应用于ACP检测。我们发现向ABPL水溶液中加入ACP后,体系的荧光强度增强,机理为:2-APBA二聚体为聚集诱导发光增强(AIEE)分子,其AIEE特性源于2-APBA二聚体的高度有序堆积;当2-APBA二聚体分子上引入PLP后(即合成ABPL),分子的高度有序堆积结构被破坏;加入ACP后,PLP水解为吡哆醛并从2-APBA二聚体分子上脱落,2-APBA二聚体分子又可重新进行高度有序堆积,表现为体系的荧光增强。在0~5 U·L^(-1)活性范围内,376.5 nm处的相对荧光强度与ACP活性之间呈现出良好的线性关系(R 2=0.99)。此外,当ACP、果胶酶、木瓜蛋白酶和脂肪酶的活性分别为4、50000、80000和10000 U·L^(-1)时,四种酶对应的相对荧光强度分别为0.2、-0.006、0.03和0.05。该结果表明ABPL对ACP具有高选择性。ABPL分子易合成,对ACP具有线性和高选择性响应,为设计合成可用于ACP检测的新型高效荧光底物提供了新策略。