Nerve growth factor pretreatment against glutamate-induced hippocampal neuronal injury Action mechanism of phosphatase and tensin homologue deleted on chromosome 10

BACKGROUND： Nerve growth factor （NGF） attenuates glutamate-induced injury to hippocampal neurons, and the human tumor suppressor gene phosphatase and tensin homologue deleted on chromosome 10 （PTEN） promotes neuronal apoptosis. However, effects of PTEN in NGF-mediated neuroprotection against glutamate excitotoxicity remain poorly understood. OBJECTIVE： To investigate the relationship between NGF inhibition of glutamate-induced injury and PTEN. DESIGN, TIME AND SE＇I＇rlNG： The randomized, controlled, in vitro study was performed at the Department of Pathophysiology, Medical School of Nantong University, China from October 2007 to March 2008. MATERIALS： Glutamate, NGF, 4, 6-diamidino-2-phenyl-indolediacetate, 3-[4, 5-dimethylthiazol-2-yl]- 2, 5-diphenyl tetrazoliumbromide （M-I-F）, and lactate dehydrogenase kit （Sigma, USA）, fluorescence microscope and inverted phase contrast microscope （Olympus, Japan） were used in this study. METHODS： Hippocampal neurons were obtained from newborn （〈 24 hours） Sprague Dawley rats and cultured for 7 days. The control group was not treated with any intervention factor, the glutamate group was treated with glutamate （0.2 mmol/L）, and NGF groups were treated with NGF （10, 50, 100, and 200 μg/L, respectively） prior to glutamate treatment. MAIN OUTCOME MEASURES： The MTT and lactate dehydrogenase assays were applied to evaluate viability of hippocampal neurons. Morphological changes in hippocampal neurons were observed using an inverted phase-contrast microscope, and neuronal apoptosis was detected by 4, 6-diamidino-2- phenyl-indolediacetate staining. PTEN mRNA and protein expression were measured by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. RESULTS： Glutamate （0.2 mmol/L） induced significantly decreased neuronal viability and greater lactate dehydrogenase efflux compared with the control group （P 〈 0.01）. However, compared with the glutamate group, cell viability significantly increased and lactate dehydrogenase efflux decreased in the NGF group with increasing NGF concentrations （P 〈 0.05 or P 〈 0.01）. The apoptotic ratio and PTEN mRNA and protein expression decreased in the NGF group compared with the glutamate group （P 〈 0.01）. CONCLUSION： Pretreatment with NGF exerted neuroprotective effects against glutamate-induced injury, partially through inhibition of PTEN expression and neuronal apoptosis.

Phosphatase and tensin homology deleted in chromosome 10,hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression

To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein expression in the patients with human colorectal adenomas and adenocarcinomas.Methods The expression of PTEN,HIF-1 alpha gene was detected by using in situ hybridization,and the VEGF expression levels by immunohistochemistry in colorectal adenomas and primary colorectal adenocarcinoma.Results Strong expression of HIF-1 alpha was detectable in the majority of colorectal dadenocarcinoma,particularly surrounding areas of necrosis in adenocarcinoma.PTEN,HIF-1 alpha mRNA and VEGF protein were positive in 51.6%,67.7% and 59.7% respectively in 62 cases of adenocarcinomas,and 77.8%,44.4% and 33.3% respectively in 18 cases of adenomas.The positive rate of VEGF was higher in the patients with colorectal adenocarcinomas than that in those with adenomas,whereas that of PTEN mRNA was contrary.HIF-1 mRNA expression was correlated significantly with lymph node metastasis,liver metastasis,Duke’s stage and recurrence.During colorectal tumor progression,the expression of HIF-1 alpha mRNA was positively correlated with the VEGF protein expression (χ2= 4.751 ,P<0.05),but negatively with the PTEN mRNA expression(χ2=21.84,P<0.01).Conclusion The absence or low expression of PTEN and the increased levels of HIF-1α and VEGF may paly an important role in carcinogenesis and progression of colorectal carcinoma.These results suggest that VEGF upregulated by HIF-1 alpha gene may be involved in angiogenesis of colorectal adenocarcinoma.4 refs,1 tab.

Rapid construction of phosphatase and tensin homolog-deleted on chromosome ten gene recombinant adenovirus using the AdEasy system

Recent studies have shown that phosphatase and tensin homolog-deleted on chromosome ten （PTEN） gene plays an important role in ischemic brain damage and synaptic plasticity. The AdEasy system, which has been widely used, greatly simplifies preparation of recombinant adenovirus. Therefore, recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene （Ad-PTEN） was constructed using the AdEasy-1 system and was transfected into HEK293 cells for packaging and amplification. Infection efficiency and expression intensity were observed in primary cultured rat hippocampal neurons infected with Ad-PTEN in vitro. Results revealed a cytopathic effect in green fluorescent protein expression, which increased with prolonged time. After three cycles of amplification, the adenovirus titer was increased to an adequate titer for infecting hippocampal neurons. The entire process typically requires 4-5 weeks for completion. Results suggested that recombinant defective adenovirus vector carrying the PTEN gene was successfully and rapidly constructed using the AdEasy system.

Hepatoprotective effects of Xiaoyao San formula on hepatic steatosis and inflammation via regulating the sex hormones metabolism

BACKGROUND The modified Xiaoyao San(MXS)formula is an adjuvant drug recommended by the National Health Commission of China for the treatment of liver cancer,which has the effect of preventing postoperative recurrence and metastasis of hepatocellular carcinoma and prolonging patient survival.However,the molecular mechanisms underlying that remain unclear.AIM To investigate the role and mechanisms of MXS in ameliorating hepatic injury,steatosis and inflammation.METHODS A choline-deficient/high-fat diet-induced rat nonalcoholic steatohepatitis(NASH)model was used to examine the effects of MXS on lipid accumulation in primary hepatocytes.Liver tissues were collected for western blotting and immunohisto chemistry(IHC)assays.Lipid accumulation and hepatic fibrosis were detected using oil red staining and Sirius red staining.The serum samples were collected for biochemical assays and NMR-based metabonomics analysis.The inflammation/lipid metabolism-related signaling and regulators in liver tissues were also detected to reveal the molecular mechanisms of MXS against NASH.RESULTS MXS showed a significant decrease in lipid accumulation and inflammatory response in hepatocytes under metabolic stress.The western blotting and IHC results indicated that MXS activated AMPK pathway but inhibited the expression of key regulators related to lipid accumulation,inflammation and hepatic fibrosis in the pathogenesis of NASH.The metabonomics analysis systemically indicated that the arachidonic acid metabolism and steroid hormone synthesis are the two main target metabolic pathways for MXS to ameliorate liver inflammation and hepatic steatosis.Mechanistically,we found that MXS protected against NASH by attenuating the sex hormone-related metabolism,especially the metabolism of male hormones.CONCLUSION MXS ameliorates inflammation and hepatic steatosis of NASH by inhibiting the metabolism of male hormones.Targeting male hormone related metabolic pathways may be the potential therapeutic approach for NASH.

Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis

AIM To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten(PTEN) gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.METHODS rat primary hepatic stellate cells(HSCs) and human LX-2 cells were transfected with adenovirus containing c DNA constructs encoding wild-type PTEN(Ad-PTEN), PTEN mutant G129 E gene(Ad-G129E), and r NA interference constructs targeting the PTEN sequence PTEN short hairpin r NA to up-regulate and downregulate the expression of PTEN. HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl_4-induced rat hepatic fibrosis model was established to investigate the in vivo effects. Hematoxylin and eosin, and Masson's trichrome were used to assess the histological changes. The expression of collagen Ⅰ and Ⅲ was assessed using immunohistochemistry and western blot analysis.RESULTS Elevated expression of PTEN gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of PTEN expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase(MMP)-13(P < 0.01) and MMP-2(P < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase(TIMP)-1(P < 0.01) and TIMP-2(P < 0.01).CONCLUSION These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.

Increased susceptibility of aging gastric mucosa to injury:The mechanisms and clinical implications

This review updates the current views on aging gastric mucosa and the mechanisms of its increased susceptibility to injury. Experimental and clinical studies indicate that gastric mucosa of aging individuals-&#x0201c;aging gastropathy&#x0201d;-has prominent structural and functional abnormalities vs young gastric mucosa. Some of these abnormalities include a partial atrophy of gastric glands, impaired mucosal defense (reduced bicarbonate and prostaglandin generation, decreased sensory innervation), increased susceptibility to injury by a variety of damaging agents such as ethanol, aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs), impaired healing of injury and reduced therapeutic efficacy of ulcer-healing drugs. Detailed analysis of the above changes indicates that the following events occur in aging gastric mucosa: reduced mucosal blood flow and impaired oxygen delivery cause hypoxia, which leads to activation of the early growth response-1 (egr-1) transcription factor. Activation of egr-1, in turn, upregulates the dual specificity phosphatase, phosphatase and tensin homologue deleted on chromosome ten (PTEN) resulting in activation of pro-apoptotic caspase-3 and caspase-9 and reduced expression of the anti-apoptosis protein, survivin. The imbalance between pro- and anti-apoptosis mediators results in increased apoptosis and increased susceptibility to injury. This paradigm has human relevance since increased expression of PTEN and reduced expression of survivin were demonstrated in gastric mucosa of aging individuals. Other potential mechanisms operating in aging gastric mucosa include reduced telomerase activity, increase in replicative cellular senescence, and reduced expression of vascular endothelial growth factor and importin-&#x003b1;-a nuclear transport protein essential for transport of transcription factors to nucleus. Aging gastropathy is an important and clinically relevant issue because of: (1) an aging world population due to prolonged life span; (2) older patients have much greater risk of gastroduodenal ulcers and gastrointestinal complications (e.g., NSAIDs-induced gastric injury) than younger patients; and (3) increased susceptibility of aging gastric mucosa to injury can be potentially reduced or reversed pharmacologically.

MiR-106b-5p Inhibits Tumor Necrosis Factor-α-induced Apoptosis by Targeting Phosphatase and Tensin Homolog Deleted on Chromosome 10 in Vascular Endothelial Cells

Background： Apoptosis of endothelial cells （ECs） plays a key role in the development of atherosclerosis and there are also evidence indicated that phosphatase and tensin homolog deleted on chromosome 10 （PTEN） is a viable target in therapeutic approaches to prevent vascular ECs apoptosis. Aberrant miR-106b-5p expression has been reported in the plasma of patients with unstable atherosclerotic plaques. However, the role and underlying mechanism of miR-106-5p in the genesis of atherosclerosis have not been addressed. In this study, we explored the anti-apoptotic role of miR-106-5p by regulating PTEN expression in vascular ECs. Methods： Real-time reverse transcription polymerase chain reaction （RT-PCR） was performed to detect the expression levels of miR-106b-5p in human atherosclerotic plaques and normal vascular tissues. Human umbilical vein endothelial cells （HUVEC） were transfected with miR-106b-5p mimic or negative control mimic, and apoptosis was induced by serum starvation and tumor necrosis factor-α （TN F-α） treat. Western blotting and real-time RT-PCR experiments were used to detect PTEN expression levels and TN F-α-induced apoptosis was evaluated by the activation of caspase-3 and cell DNA fragmentation levels in HUVEC. Results： The expression ofmiR-106b-5p was significantly downregulated in plaques than in normal vascular tissues. TNF-α significantly downregulated miR-106b-5p expression levels and upregulated activation of caspase-3 and cell DNA fragmentation levels in HUVEC. Overexpression ofmiR-106b-5p with miR-106b-5p mimic inhibited PTEN expression and TNF-α-induced apoptosis in HUVEC. Luciferase reporter assays confirmed that miR-106b-5p binds to PTEN mRNA 3＇ untranslated region site, Conclusion： MiR-106b-5p could inhibit the expression of PTEN in vascular ECs, which could block TNF-α-induced activation of caspase-3, thus prevent ECs apoptosis in atherosclerosis diseases.

PTEN and Ki67 expression is associated with clinicopathologic features of non-small cell lung cancer

Phosphatase and tensin homolog deleted on chromosome 10（PTEN） and the proliferating antigen Ki67 have been widely studied in several tumors.However,their role as indicator in non-small cell lung cancer（NSCLC）remains unknown.Here,we investigated the expression of PTEN and Ki67 in NSCLC tissues and paired normal lung tissues to identify whether these proteins are associated with lung cancer development and survival.Immunohistochemistry for PTEN and Ki67 was performed on 67 lung cancer tissues and 41 paired adjacent normal lung tissues to detect the expression of these two proteins.The expression of PTEN in NSCLC tissues（32.8%） was significantly lower than that in normal tissues（82.9%,P 〈 0.05）.In contrast,the expression of Ki67 in NSCLC tissues（76.1%） was significantly higher than that in normal tissues（27.3%,P 〈 0.05）.Expression of both PTEN and Ki67 were strongly associated with tumor histology,clinical stage,lymph node metastasis,differentiation and4-year postoperative survival rate（P 〈 0.05）.However,PTEN expression was negatively correlated with Ki67 expression（r =-0.279,P 〈 0.05）.In conclusion,low PTEN expression and Ki67 overexpression are associated with malignant invasion and lymph node metastasis of NSCLC.These proteins may serve as diagnostic and prognostic biomarkers of NSCLC.

MiR-200a and miR-200b target PTEN to regulate the endometrial cancer cell growth in vitro

Objective:To study whether miR-200a and miR-200b target PTEN gene expression to regulate the endometrial cancer cell growth in vitro. Methods:Endometrial cancer cells ECC-1 were cultured and transfected with the miR-200a and miR-200b mimics and inhibitors as well as the negative control mimics and inhibitors,and then the cell proliferation activity as well as the expression of PTEN and downstream genes in cells was determined; after transfection of miR-200a and miR-200b mimics as well as PTEN-3'UTR luciferase report gene plasmids,the fluorescence activity of luciferase reporter gene was determined. Results:12 h,24 h and 48 h after transfection,the cell proliferation activity of miR-200a mimics group and miR-200b mimics group were significantly higher than those of NC mimics group while the cell proliferation activity of mi R-200 a inhibitor group and miR-200b inhibitor group were significantly lower than those of NC inhibitor group; 48 h after transfection,PTEN expression in cells and PTEN-3'UTR luciferase reporter gene fluorescence activity of miR-200 a mimics group and miR-200b mimics group were significantly lower than those of NC mimics group while p-PI3K and p-Akt expression were significantly higher than those of NC mimics group; PTEN expression in cells and PTEN-3'UTR luciferase reporter gene fluorescence activity of miR-200 inhibitor group and miR-200b inhibitor group were significantly higher than those of NC inhibitor group while p-PI3K and p-Akt expression were significantly lower than those of NC inhibitor group. Conclusion:miR-200 a and miR-200b can promote the endometrial cancer cell growth in vitro by targeted inhibition of PTEN gene expression.

Upregulated DJ-1 Promotes Renal Tubular EMT by Suppressing Cytoplasmic PTEN Expression and Akt Activation

Recently,phosphatase and tensin homolog deleted on chromosome 10（PTEN） is suggested as a new agent in the fighting against fibrogenesis.In tumor,DJ-1 is identified as a negative regulator of PTEN.But the expression of DJ-1 and the regulation of PTEN in fibrosis are unclear.Renal fibrosis was induced in 5/6 subtotal nephrectomy rat model.Human proximal tubular epithelial cells（HKC） were treated with transforming growth factor-beta 1（TGF-β1）,or transfected with DJ-1 or PTEN.Confocal microscope was used to investigate the localization of DJ-1 and PTEN.The selective phosphoinositide-3 kinase（PI3K） inhibitor,LY294002,was administered to inhibit PI3K pathway.The DJ-1 and PTEN expression,markers of epithelial-mesenchymal transition（EMT） and Akt phosphorylation were measured by RT-PCR,Western blotting or immunocytochemistry.In vitro,after HKC cells were stimulated with 10 ng/mL TGF-β1 for 72 h,the expression of DJ-1 was increased,and that of PTEN was decreased.In vivo,the same results were identified in 5/6-nephrectomized rats.In normal HKC cells,most of DJ-1 protein localized in cytoplasm,and little in nucleus.TGF-β1 upregulated DJ-1 expression in both cytoplasma and nuclei.In contrary,TGF-β1 emptied cytoplasmic PTEN protein into nucleus.Overexpression of DJ-1 decreased the expression of PTEN,promoted the activation of Akt and the expression of vimentin,and also led to the loss of cytoplasmic PTEN.Contrarily,overexpression of PTEN protected HKC cells from TGF-β1-induced EMT.In conclusion,DJ-1 is upregulated in renal fibrosis and DJ-1 mediates EMT by suppressing cytoplasmic PTEN expression and Akt activation.

Relationsip between PTEN and VEGF Expression and Clinicopathological Characteristics in HCC

To investigate the expressions and significance of the tumor suppressor gene phosphatase and tensin homlog deleted on chromosome ten protein （PTEN） and vascular endothelial growth factor （VEGF） in hepatocellular carcinoma （HCC）, and to analyze the relationship between their expressions and the tumor＇s invasion and their pericarcinomatous tissues, the correlation of their expressions with the tumor＇s clinicopathological characteristics and invasion potential were studied. Our study showed that the expression level of PTEN in HCC was remarkably lower than that in pericarcinomatous liver tissues, while the expressions of both VEGF and MVD were higher than that in pericarcinomatous liver tissues. Correlation analysis revealed that the expression of PTEN was negatively related to the progression of the pathological differentiation and invasion of tumor, whereas the expressions of VEGF and MVD were positively related. Moreover, there was a negative relationship between the expression of PTEN and the expressions of VEGF and MVD, and a positive one between VEGF and MVD. The expressions of PTEN and VEGF may reveal the degree of differentiation and the invasive potential of HCC tissues. The mechanism by which the lack of PTEN expression probably induces abnormal hyperexpression of VEGF may play an important role in the invasion and metastasis of HCC.

IL-37调控miR-106b-5p/PTEN抑制肾细胞癌细胞生物学行为

目的:探讨IL-37对肾细胞癌细胞生物学行为的影响和潜在机制。方法:RT-qPCR检测肾细胞癌组织中IL-37mRNA和miR-106b-5p表达。Western blot检测肾细胞癌组织中10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)蛋白表达。将肾细胞癌细胞786-0分为对照组、IL-37(10、50、100 ng/ml)组、anti-miR-NC组、anti-miR-106b-5p组、IL-37+miR-NC组、IL-37+miR-106b-5p组。采用MTT法、平板克隆实验、划痕愈合实验、流式细胞术检测786-0细胞增殖、迁移和凋亡能力。荧光素酶实验检测miR-106b-5p与PTEN的靶向关系。结果:肾细胞癌组织中miR-106b-5p表达量显著升高(P<0.05),IL-37 mRNA和PTEN蛋白表达量显著降低(P<0.05)。与对照组比较,IL-37(10、50、100 ng/ml)组细胞活力、集落形成数、迁移距离、miR-106b-5p表达显著降低(P<0.05),凋亡率、PTEN蛋白表达显著升高(P<0.05)。与anti-miR-NC组比较,anti-miR-106b-5p组细胞活力、集落形成数、迁移距离显著降低(P<0.05),凋亡率、PTEN蛋白表达显著升高(P<0.05)。与IL-37+miR-NC组比较,IL-37+miR-106b-5p组细胞活力、集落形成数、迁移距离显著升高(P<0.05),凋亡率、PTEN蛋白表达显著降低(P<0.05)。PTEN是miR-106b-5p的靶基因。结论:外源性IL-37可抑制肾细胞癌细胞的增殖和迁移,诱导细胞凋亡,其抗肿瘤机制可能是通过抑制miR-106b-5p/PTEN途径发挥作用。

微RNA-132-3p靶向第10号染色体缺失的磷酸酶和张力蛋白同源物调控葡萄膜黑色素瘤细胞增殖与凋亡

目的 研究微RNA(miR)-132-3p在葡萄膜黑色素瘤细胞增殖、凋亡中的作用及其作用机制。方法 该研究起止时间为2018年2月至2019年7月。定量聚合酶链反应(qPCR)检测正常葡萄膜上皮细胞ARPE-19和葡萄膜黑色素瘤细胞SP6.5、M23中miR-132-3p表达。SP6.5细胞中转染miR-132-3p干扰质粒(anti-miR-132-3p)、第10号染色体上缺失的磷酸酶和张力蛋白同源物(PTEN)过表达质粒(pcDNA3.1-PTEN)或共转染anti-miR-132-3p和PTEN干扰质粒(si-PTEN),MTT法和流式细胞术分别检测细胞增殖与凋亡,蛋白质印迹法检测PTEN、细胞周期蛋白D1(cyclin D1)、周期素依赖激酶抑制剂p21(P21)、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白表达,生物信息学预测结合双萤光素酶报告实验分析miR-132-3p与PTEN的靶向关系。结果 与ARPE-19细胞相比,SP6.5、M23细胞中miR-132-3p表达量(0.26±0.02比0.94±0.09、0.81±0.08)明显升高(P<0.05)。与anti-miR-132-3p阴性对照(anti-miR-NC)组相比,anti-miR-132-3p组24 h、48 h、72 h的细胞活性(0.51±0.05比0.30±0.03、0.97±0.09比0.45±0.05、1.40±0.14比0.76±0.07)、cyclin D1、Bcl-2蛋白表达量显著降低(P<0.05),细胞凋亡率[(8.03±0.68)%比(21.51±2.06)%]、P21、Bax水平明显提高(P<0.05),与过表达PTEN相同。miR-132-3p与PTEN之间有靶向调控关系。抑制PTEN能逆转抑制miR-132-3p对SP6.5细胞增殖的抑制作用及对细胞凋亡的促进作用。结论 miR-132-3p通过直接靶向PTEN调控葡萄膜黑色素瘤细胞增殖与凋亡。

间充质干细胞外泌体对缺血性脑卒中大鼠神经功能恢复的影响

目的探讨间充质干细胞外泌体(MSCs-EXO)对缺血性脑卒中大鼠神经功能恢复的影响。方法大鼠骨髓间充质干细胞原代培养,超速离心法提取其MSCs-EXO,大脑中动脉夹闭模型(MCAO)法制作大鼠缺血性脑卒中模型,MSCs-EXO经鼻给药,设为MSCs-EXO组,通过改良神经功能缺损评分(mNSS)和错步试验评估其神经功能恢复情况,并与未治疗模型组(MCAO组)做对比。PKH26标记MSCs-EXO并结合免疫荧光染色观察其在缺血半暗带(IP)中的分布,荧光定量PCR及Western blot检测IP区域脑组织中PTEN的表达水平,并与正常大鼠及MCAO组大鼠进行对比。结果MSCs-EXO治疗组的神经功能恢复高于MCAO组,差异有统计学意义(P<0.05)。免疫荧光染色显示标记外泌体可以进入IP区域神经细胞,荧光定量PCR及Western blot检测显示MSCs-EXO治疗组及MCAO组中PTEN水平均较正常升高,但MSCs-EXO治疗组的PTEN水平低于MCAO组,差异有统计学意义(P<0.05)。结论MSCs-EXO能够促进缺血性脑卒中大鼠的神经功能恢复,这可能与其下调IP区域PTEN表达水平相关。

自身免疫性肝病患者血清PRDX1、PTEN水平及其与肝功能、疾病活动性的关系

目的探讨过氧化物氧化还原蛋白(PRDX)1、第10号染色体缺失性磷酸酶-张力蛋白同源物基因(PTEN)水平与自身免疫性肝病患者肝功能、疾病活动性的关系。方法选取2021年1月至2022年12月该院收治的83例自身免疫性肝病患者作为研究对象,根据入院时疾病活动性分为活动期组(37例)、缓解期组(46例),统计两组临床资料及入院时血清PRDX1、PTEN水平,同时对患者进行肝功能Child-Pugh分级并分组。选取同期体检的100例健康志愿者作为对照组。采用多因素Logistic逐步回归分析自身免疫性肝病患者疾病活动性的影响因素,采用受试者工作特征(ROC)曲线及曲线下面积(AUC)分析治疗后血清PRDX1、PTEN水平对自身免疫性肝病患者疾病活动性的评估价值。结果与A级组比较,B级组血清PRDX1、PTEN水平差异无统计学意义(P>0.05),而C级组血清PRDX1水平升高,PTEN水平降低(P<0.05);与B级组相比,C级组血清PRDX1水平升高、PTEN水平降低(P<0.05);与对照组比较,缓解期组血清PRDX1、PTEN水平差异无统计学意义(P>0.05),而活动期组血清PRDX1水平升高、PTEN水平降低(P<0.05);与缓解期组相比,活动期组血清PRDX1水平升高、PTEN水平降低(P<0.05)。血清PRDX1、PTEN判断自身免疫性肝病患者疾病活动性的AUC分别为0.750、0.854,二者联合预测的AUC为0.916。活动期组患者肝区不适、肝硬化占比高于缓解期组(P<0.05);多因素Logistic逐步回归分析显示,肝区不适(OR=3.487,95%CI:1.534~7.927),肝硬化(OR=4.289,95%CI:1.744~10.545),PRDX1≥5.22 ng/mL(OR=5.068,95%CI:1.951~13.164),PTEN≤0.31 pg/mL(OR=5.387,95%CI:2.099~13.829)是影响自身免疫性肝病疾病活动性的危险因素(P<0.05)。结论血清PRDX1水平升高、PTEN水平降低与自身免疫性肝病患者肝功能、疾病活动性密切相关,二者对自身免疫性肝病患者具有一定临床评估价值。

PTEN、CA125、sVEGFR1、NGAL在子宫内膜癌患者血清中的表达及与病理特征的关系

目的 探讨第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)、糖类抗原125(CA125)、可溶性血管内皮生长因子受体-1(sVEGFR1)、中性粒细胞明胶酶相关脂质载运蛋白(NGAL)在子宫内膜癌(EC)患者血清中的表达及与病理特征的关系。方法 选取2019年1月至2021年6月于在邯郸市第一医院行全子宫切除术并经病理诊断的120例EC患者为研究组,选择同期80例良性病变子宫内膜患者为对照组,用酶联免疫吸附法检测并比较2组患者血清PTEN、CA125、sVEGFR1、NGAL水平,收集2组临床病理资料,分析血清PTEN、CA125、sVEGFR1、NGAL与研究组患者病理特征的关系。结果 研究组血清CA125、NGAL水平高于对照组,血清PTEN、sVEGFR1水平低于对照组(P<0.05)。分化程度越低血清CA125、NGAL水平越高,血清PTEN、sVEGFR1水平越低(P<0.05);临床分期Ⅲ~Ⅳ期患者血清PTEN高于Ⅰ~Ⅱ期(P<0.05);临床分期Ⅲ~Ⅳ期、有淋巴结转移、浸润深度≥50%患者血清CA125、NGAL水平升高,血清sVEGFR1水平降低(P<0.05)。血清PTEN与临床分期呈负相关,与分化程度呈正相关(P<0.05);血清CA125、NGAL与临床分期、淋巴结转移、浸润深度呈正相关,与分化程度呈负相关(P<0.05);血清sVEGFR1与临床分期、淋巴结转移、浸润深度呈负相关,与分化程度呈正相关(P<0.05)。结论 CA125、NGAL在EC患者血清中呈高表达,PTEN、sVEGFR1呈低表达,均与EC患者病理特征有一定相关性,可作为EC早期诊断与疾病进展的潜在生物学标志物。

GATA3介导miR-21/PTEN轴对子宫内膜癌细胞增殖、侵袭的影响

目的分析GATA结合蛋白3(GATA3)介导微小RNA-21(miR-21)/人类第10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)轴对子宫内膜癌细胞增殖、侵袭的影响。方法取HEC-1-A细胞,进行转染分组,分为对照组、GATA3空载质粒组、GATA3过表达质粒组、GATA3 siRNA阴性对照组、GATA3 siRNA组。检测各组细胞中GATA3、miR-21、PTEN表达量、增殖情况、凋亡率、迁移、侵袭。结果与hEEC组相比,HEC-1-A组、HEC-1-B组、Ishikawa组细胞中GATA3、miR-21表达水平升高,PTEN表达水平降低(P<0.05)。与GATA3空载质粒组相比,GATA3过表达质粒组GATA3、miR-21 mRNA表达量、增殖率、迁移距离、侵袭细胞数、Vimentin水平升高,PTEN mRNA表达量、凋亡率、Caspase-9、Bax、E-cadherin水平降低(P<0.05);与GATA3 siRNA阴性对照组相比,GATA3、miR-21 mRNA表达量、增殖率、迁移距离、侵袭细胞数、Vimentin水平降低,PTEN mRNA表达量、凋亡率、Caspase-9、Bax、E-cadherin水平升高(P<0.05)。结论下调GATA3表达,可对miR-21/PTEN轴进行调节,使HEC-1-A细胞的增殖减慢,促进HEC-1-A细胞的凋亡。

PTEN基因在鸡肝脏中的发育性变化

为了研究抑癌基因PTEN在家禽肝脏发育过程中的表达变化,试验选取50枚海兰褐受精蛋,在孵化的第15,18,20天[E15、E18、E20(啄壳为准)]和出壳后第1,5,10,15天(D1、D5、D10、D15)各取6枚(只)鸡胚/雏鸡,用乙醚麻醉后颈椎脱臼致死,取肝脏并提取基因组,采用实时荧光定量PCR和免疫组织化学方法对PTEN基因在家禽肝脏发育过程中的表达变化进行研究。结果表明:PTEN基因相对表达量随着胚龄/日龄的增长而增加,但在啄壳当天,PTEN基因相对表达量会暂时下降。PTEN蛋白在出壳前期主要分布在部分肝血窦内皮细胞的细胞核和肝细胞的细胞核中,在细胞质中基本无表达;在出壳后,PTEN蛋白在肝细胞核和细胞浆内均有分布。E20,免疫阳性PTEN蛋白的表达量下降,和E15水平相当(P>0.05),在其他胚龄/日龄时均显著高于E15(P<0.05)。肝细胞核PTEN蛋白免疫阳性率在E18显著高于E15(P<0.05);在E20和D15,肝细胞核PTEN蛋白免疫阳性率急剧下降,均显著低于E18(P<0.05);在D5~D15,肝细胞核PTEN蛋白免疫阳性率逐渐增加,均显著高于E15(P<0.05)。PTEN基因的这种表达模式和鸡胚/雏鸡肝脏的发育模式基本一致,说明家禽PTEN基因在肝脏发育和代谢中发挥重要作用。

伴微量白蛋白尿2型糖尿病患者血清脂肪细胞型脂肪酸结合蛋白和4和第10号染色体缺失的磷酸酶张力蛋白同源物蛋白的研究

目的探讨2型糖尿病(T2DM)伴微量白蛋白尿患者血清脂肪细胞型脂肪酸结合蛋白(FABP4)和第10号染色体缺失的磷酸酶和张力蛋白同源物(PTEN)蛋白表达水平变化及两者在糖尿病肾病(DN)发生过程中的相互关系。方法收集T2DM患者120例,据尿白蛋白肌酐比值(UACR)进行分组,其中正常白蛋白尿组(D0)39例,微量白蛋白尿组(D1)81例。同时收集39例正常对照组(NC)。采用ELI-SA法检测受试者外周血清FABP4和PTEN蛋白表达的水平。结果 T2DM组血清FABP4和PTEN蛋白水平均显著高于正常对照组(P <0.001)。D1组血清FABP4和PTEN蛋白水平显著高于NC组及D0组(均P <0.05)。T2DM患者中,Log(FABP4)与PTEN蛋白(r=0.524,P <0.001)、Log(UACR)(r=0.202,P <0.05)均呈正相关。二分类Logistic回归分析显示,血清FABP4水平与T2DM患者尿微量白蛋白的出现独立相关(OR=1.147,95%CI∶1.042～1.263,P=0.005)。结论血清FABP4水平也许可作为DN患者的早期预测指标。FABP4和PTEN蛋白可能在DN的发生中存在着相互联系。

血清TLR4和PTEN与特发性膜性肾病患者临床病理特征及预后的相关性

目的 探究血清Toll样受体4(TLR4)、第10号染色体缺失性磷酸酶和张力蛋白同源物基因(PTEN)与特发性膜性肾病(IMN)患者临床病理特征及预后的相关性。方法 选取2019年1月至2022年1月在河北以岭医院就诊治疗的IMN患者224例为IMN组,同时根据其预后结果将其分为预后良好组174例、预后不良组50例;另选取同期在本院体检健康者100名作为对照组。采用酶联免疫吸附法(ELISA)检测研究对象血清TLR4、PTEN水平;采用pearson分析IMN患者血清TLR4水平和PTEN水平相关性;多因素Logistic回归分析IMN患者预后不良的影响因素;受试者工作特征曲线(ROC)分析血清TLR4和PTEN在评估IMN患者预后中的价值。结果 IMN组血清TLR4、PTEN水平较对照组均显著升高,差异有统计学意义(t=12.918,13.249,P<0.05);预后不良组血清TLR4、PTEN水平较预后良好组均显著升高,差异有统计学意义(t=11.608,12.965,P<0.05);IMN患者血清TLR4与PTEN水平呈正相关;TLR4、PTEN、IgG、合并高血压均为IMN患者预后不良的独立危险因素(P<0.05);血清TLR4、PTEN水平预测IMN患者预后是否不良的曲线下面积(AUC)分别为0.878、0.868,两者联合预测的AUC为0.941,高于两者单独预测(z=1.874、2.065,P<0.05),且特异度为0.89,灵敏度为0.88。结论 血清TLR4、PTEN水平与IMN患者分期,肾小球硬化,二者对IMN预后具有一定的预测价值。