Current concepts in ameloblastoma-targeted therapies in B-raf proto-oncogene serine/threonine kinase V600E mutation: Systematic review

BACKGROUND Ameloblastomas are common benign epithelial odontogenic neoplasms that present an aggressive and unpredictable behavior that may modify treatment strategies.Different signaling pathways that participate in the progression of these tumors have been identified.B-raf proto-oncogene serine/threonine kinase(BRAF)is a protein involved in the behavior of ameloblastomas,and it is related to many cell mechanisms.BRAF gene mutations have been identified in ameloblastomas,of which the BRAF V600E(valine substituted by glutamic acid at amino acid 600)mutation has been the most common and can be present concomitantly with other mutations that may be involved in its behavior.Targeted therapies have been used as an alternative in the case of resistance or contraindications to conventional treatments.AIM To document the presence of BRAF V600E and additional mutations,their behavior,and targeted therapies in these tumors.METHODS An electronic literature search was conducted according to PRISMA guidelines in PubMed/MEDLINE,Cochrane,EMBASE,and SpringerLink using the terms“ameloblastomas”,“BRAF V600E”,“additional mutations”,and“targeted therapies”.Ameloblastomas were classified according to WHO guidelines.Inclusion criteria were articles in English,published not more than 10 years ago,and studies with laboratory works related to BRAF V600E.Articles were evaluated by two independent reviewers and retrieved for full-text evaluation.The EBLIP Critical Appraisal Checklist was used to evaluate the quality of the eligible studies.Descriptive statistical analysis was performed.RESULTS Two independent reviewers,with a substantial concordance indicated by a kappa coefficient of k=0.76,evaluated a total of 19 articles that were included in this study.The analysis registered 521 conventional ameloblastomas(AM),81 unicystic ameloblastomas(UA),13 ameloblastic carcinomas(AC),three metastatic ameloblastomas(MA),and six peripheral ameloblastomas(PA),of which the histopathological type,anatomic location,laboratory tests,expression of BRAF mutation,and additional mutations were registered.The BRAF V600E mutation was found in 297 AM(57%),63 UA(77.7%),3 AC(23%),1 MA(50%),and 5 PA(83.3%).Follicular type predominated with a total of 116 cases(40%),followed by plexiform type with 63 cases(22.1%).Furthermore,both types presented additional mutations,in which alterations in JAK3 P132T,SMARCB1,PIK3CA,CTNNB1,SMO,and BRAF G606E genes were found.Four case reports were found with targeted therapy to BRAF V600E.CONCLUSION The identification of BRAF V600E and additional mutations as an aid in targeted therapies has been a breakthrough in alternative treatments of ameloblastomas where surgical treatments are contraindicated.

Novel serine/threonine kinase 11 gene mutations in PeutzJeghers syndrome patients and endoscopic management

AIM:To explore mutations in serine/threonine kinase 11(STK11) gene in Peutz-Jeghers syndrome(PJS) with gastrointestinal(GI) hamartomatous polyps.METHODS:Six Japanese PJS patients in 3 families were enrolled in this study.Each of the cases had hamartomatous polyposis in the gastrointestinal tract,including the small intestine,along with mucocutaneous hyperpigmentation.Narrow-band imaging(NBI)-magnification endoscopy was employed to detect microvascular and microsurface irregularities in the GI lesions.NBI magnification findings could be classified into three groups(type A,type B,or type C).Endoscopic polypectomy was performed using double-balloon enteroscopy or colonoscopy.Genomic DNA was extracted from a whole blood sample from each subject.All of the coding exons of STK11 gene,its boundary regions,and the promoter region containing the polymorphic regions were amplified by polymerase chain reaction,and direct sequencing was performed to assess the germline mutations.RESULTS:NBI-magnification endoscopic observation could detect the abnormalities in microvessels and microsurface structures of GI polyps.Overall,we found 5 cases of type A and one case without the examination for the gastric polyps,while there were 4 cases of type B and 2 case of type A for the colorectal polyps.Seventy-nine small-bowel and 115 colorectal polyps over 27 sessions for each were resected endoscopically without significant complications.The only delayed complication included the occurrence of bleeding in a case,and this was successfully managed with hemoclips.Resected polyps contained no malignant components.Based on mutation analysis,all 3 cases in Family I exhibited the +658C>T nonsense mutation in exon 5,which resulted in the production of a truncated protein(Q220X).In Family II,a case had-252C>A and-193C>A in the promoter region.In Family III,a case was found to have the +1062C>G(F342L) mutation in exon 8.CONCLUSION:We found two novel mutations of STK11 in association with PJS.Endoscopic polypectomy of GI polyps in PJS patients appears to be useful to prevent emergency laparotomies and reduce the cancer risk.

Serine-threonine protein kinase activation may be an effective target for reducing neuronal apoptosis after spinal cord injury

The signaling mechanisms underlying ischemia-induced nerve cell apoptosis are poorly understood. We investigated the effects of apoptosis-related signal transduction pathways following ischemic spinal cord injury, including extracellular signal-regulated kinase（ERK）, serine-threonine protein kinase（Akt） and c-Jun N-terminal kinase（JNK） signaling pathways. We established a rat model of acute spinal cord injury by inserting a catheter balloon in the left subclavian artery for 25 minutes. Rat models exhibited notable hindlimb dysfunction. Apoptotic cells were abundant in the anterior horn and central canal of the spinal cord. The number of apoptotic neurons was highest 48 hours post injury. The expression of phosphorylated Akt（pAkt） and phosphorylated ERK（p-ERK） increased immediately after reperfusion, peaked at 4 hours（p-Akt） or 2 hours（p-ERK）, decreased at 12 hours, and then increased at 24 hours. Phosphorylated JNK expression reduced after reperfusion, increased at 12 hours to near normal levels, and then showed a downward trend at 24 hours. Pearson linear correlation analysis also demonstrated that the number of apoptotic cells negatively correlated with p-Akt expression. These findings suggest that activation of Akt may be a key contributing factor in the delay of neuronal apoptosis after spinal cord ischemia, particularly at the stage of reperfusion, and thus may be a target for neuronal protection and reduction of neuronal apoptosis after spinal cord injury.

Tacolimus Postconditioning Alleviates Apoptotic Cell Death in Rats after Spinal Cord Ischemia-reperfusion Injury via Up-regulating Protein-Serine-Threonine Kinases Phosphorylation

The effects of tacrolimus postconditioning on protein-serine-threonine kinases （Akt） phos- phorylation and apoptotic cell death in rats after spinal cord ischemia-reperfusion injury were investi- gated. Ninety male SD rats were randomly divided into sham operation group, ischemia-reperfusion group and tacrolimus postconditioning group. The model of spinal cord ischemia was established by means of catheterization through femoral artery and balloon dilatation. The spinal cord was reperfused 20 min after ischemia via removing saline out of balloon. The corresponding spinal cord segments were excised and determined for Akt activity in spinal cord tissue by using Western blotting at 5, 15, and 60 min after reperfusion respectively. Spinal cord tissue sections were stained immunohistochemically for detection of the phosphorylated Akt expression at 15 min after reperfusion. Flow cytometry was applied to assess apoptosis of neural cells, and dry-wet weights method was employed to measure water content in spinal cord tissue at 24 h after reperfusion. The results showed that the activities of Akt in tarcolimus postconditioning group were significantly higher than those in ischemia-reperfusion group at 5, 15, and 60 min after reperfusion （P〈0.05, P〈0.01）. The Akt activities reached the peak at 15 min after reperfu- sion in ischemia-reperfusion group and tacrolimus postconditioning group. The percentage of apoptotic cells and water content in spinal cord tissue were significantly reduced （P〈0.01） in tacrolimus postcon- ditioning group as compared with those in ischemia-reperfusion group at 24 h after reperfusion. It is concluded that tacrolimus postconditioning can increase Akt activity in spinal cord tissue of rats, inhibit apoptosis of neural cells as well as tissue edema, and thereby alleviate spinal cord ischemia-reperfusion injury.

Knockdown of Microtubule Associated Serine/threonine Kinase Like Expression Inhibits Gastric Cancer Cell Growth and Induces Apoptosis by Activation of ERK1/2 and Inactivation of NF-κB Signaling

Microtubule-associated serine/threonine kinase(MASTL)functions to regulate chromosome condensation and mitotic progression.Therefore,aberrant MASTL expression is commonly implicated in various human cancers.This study analyzed MASTL expression in gastric cancer vs.adjacent normal tissue for elucidating the association with clinicopathological data from patients.This work was then extended to investigate the effects of MASTL knockdown on tumor cells in vitro.The level of MASTL expression in gastric cancer tissue was assessed from the UALCAN,GEPIA,and Oncomine online databases.Lentivirus carrying MASTL or negative control shRNA was infected into gastric cancer cells.RT-qPCR,Western blotting,cell viability,cell counting,flow cytometric apoptosis and cell cycle,and colony formation assays were performed.MASTL was upregulated in gastric cancer tissue compared to the adjacent normal tissue,and the MASTL expression was associated with advanced tumor stage,Helicobacter pylori infection and histological subtypes.On the other hand,knockdown of MASTL expression significantly reduced tumor cell viability and proliferation,and arrested cell cycle at G2/M stage but promoted tumor cells to undergo apoptosis.At protein level,knockdown of MASTL expression enhanced levels of cleaved PARP1,cleaved caspase-3,Bax and p-ERK1/2 expression,but downregulated expression levels of BCL-2 and p-NF-κB-p65 protein in AGS and MGC-803 cells.MASTL overexpression in gastric cancer tissue may be associated with gastric cancer development and progression,whereas knockdown of MASTL expression reduces tumor cell proliferation and induces apoptosis.Further study will evaluate MASTL as a potential target of gastric cancer therapeutic strategy.

serine/threonine蛋白激酶功能研究进展

蛋白激酶(protein kinase)具有将ATP的γ-磷酸基转移到蛋白质底物特定的氨基酸残基上的潜在催化能力,从而使蛋白质磷酸化。根据底物上氨基酸的特异性,蛋白激酶可以细分为丝/苏氨酸激酶和酪氨酸激酶。在DNA复制和有丝分裂过程中蛋白激酶起调节作用。同时,蛋白激酶在转录过程也起到重要作用,如在转录因子核转位过程的作用、调节转录因子与DNA结合能力、调节转录因子的激活活性。蛋白激酶的磷酸化作用与肿瘤发生关系密切,其可以促使基因表达的改变等一系列细胞的应答发生,最终导致癌症的发生和发展。

Pathophysiological roles of Pim-3 kinase in pancreatic cancer development and progression

Pim-3 is a member of the provirus integration site for Moloney murine leukemia virus(Pim)family proteins that exhibit serine/threonine kinase activity.Similar to the other Pim kinases(Pim-1 and Pim-2),Pim-3 is involved in many cellular processes,including cell proliferation,survival,and protein synthesis.Although Pim-3is expressed in normal vital organs,it is overexpressed particularly in tumor tissues of endoderm-derived organs,including the liver,pancreas,and colon.Silencing of Pim-3 expression can retard in vitro cell proliferation of hepatocellular,pancreatic,and colon carcinoma cell lines by promoting cell apoptosis.Pim-3 lacks the regulatory domains similarly as Pim-1 and Pim-2 lack,and therefore,Pim-3 can exhibit its kinase activity once it is expressed.Pim-3 expression is regulated at transcriptional and post-transcriptional levels by transcription factors(e.g.,Ets-1)and post-translational modifiers(e.g.,translationally-controlled tumor protein),respectively.Pim-3 could promote growth and angiogenesis of human pancreatic cancer cells in vivo in an orthotopic nude mouse model.Furthermore,a Pim-3 kinase inhibitor inhibited cell proliferation when human pancreatic cancer cells were injected into nude mice,without inducing any major adverse effects.Thus,Pim-3 kinase may serve as a novel molecular target for developing targeting drugs against pancreatic and other types of cancer.

A defect in the PINOID serine/threonine kinase affects leaf shape in cucumber

Examining the plants in any forest or meadow reveals a remarkable diversity of leaf shape,suggesting the importance of this trait for adaptation to various environmental conditions(reviewed in Nicotra et al.2011).Indeed,leaf shape may be constrained by biomechanical factors and affects thermoregulation,susceptibility to herbivory,the available light for photosynthesis,and water balance.

LRRK2基因R1067Q和GBA基因R202Q双变异致早发型帕金森病一例

患者男性,42岁。主因左手抖动18个月,左下肢抖动伴运动迟缓6个月,于2023年7月8日入院。患者18个月前(2022年1月)无明显诱因出现左上肢不自主抖动,静止时出现、持物及动作时消失,无动作迟缓、反应变慢等其他伴随症状。于2022年11月至外院就诊,头部MRI检查无明显异常,结合临床症状,考虑帕金森病(PD),服用普拉克索0.375 mg/d并逐渐增量至0.375 mg/次、2次/d长期治疗,症状无明显改善。6个月前(2023年1月)出现左下肢不自主抖动,并逐渐出现动作迟缓,左侧肢体活动不灵活,体位变化或行走时偶有头晕,不伴视物旋转及恶心呕吐等,持续数分钟后头晕可自行好转,无嗅觉丧失、情绪低落,睡眠质量尚可,无多梦、呓语、乱喊乱叫、手舞足蹈等症状。

PIM1基因对急性髓系白血病U937细胞增殖、凋亡及JAK2/STAT3信号通路的影响

目的:探讨PIM1基因对急性髓系白血病(AML)U937细胞增殖、凋亡的影响,以及对JAK2/STAT3通路的调控作用。方法:收集初诊成人AML患者和单纯缺铁性贫血患者的骨髓单个核细胞,荧光定量PCR检测PIM1 mRNA表达。将AML细胞系U937细胞分为:U937组(U937细胞正常培养)、Si-PIM1组(U937细胞转染含PIM1 mRNA的低表达腺病毒载体)、Si-NC组(U937细胞转染不含PIM1 mRNA的低表达腺病毒载体)、CoA1组(U937细胞中加入浓度为20μmol/L的JAK2激活剂CoA1)、Si-PIM1+CoA1组(U937细胞转染含PIM1 mRNA低表达的腺病毒载体并加入浓度为20μmol/L的CoA1)。培养24 h。荧光定量PCR和蛋白印迹法检测U937细胞PIM1 mRNA和蛋白、JAK2/STAT3通路、细胞周期、凋亡相关蛋白表达;噻唑蓝法检测细胞增殖活性;流式细胞术检测细胞周期变化及凋亡率。结果:AML患者骨髓单个核细胞中PIM1 mRNA表达水平高于单纯缺铁性贫血患者(P<0.05)。与U937组相比,Si-PIM1组细胞PIM1 mRNA和蛋白、p-JAK2/JAK2、p-STAT3/STAT3、Cyclin D1、CDK2蛋白、细胞增殖活性、S期比例、G2/M期比例降低(均P<0.05),p27、Caspase-3蛋白、G0/G1期、凋亡率升高(均P<0.05),而CoA1组上述指标的变化情况与Si-PIM1组正好相反,CoA1可逆转Si-PIM1对U937细胞的作用效果。U937组、Si-PIM1+CoA1组、Si-NC组U937细胞上述指标差异无统计学意义(P>0.05)。结论:敲低PIM1基因表达可抑制U937细胞增殖、促进凋亡,缓解ALM进程,且上述作用可能与抑制JAK2/STAT3通路活化有关。

基于PI3K/Akt/mTOR 通路探究七氟醚麻醉对大鼠神经细胞凋亡的影响

目的探讨七氟醚麻醉对大鼠认知功能及神经细胞凋亡的影响。方法取96只大鼠,随机分为对照组(吸入2 L·min^(−1)氧气)、七氟醚低浓度组(吸入体积分数为1%的七氟醚+2 L·min^(−1)氧气)、七氟醚中浓度组(吸入体积分数为2%的七氟醚+2 L·min^(−1)氧气)、七氟醚高浓度组(吸入体积分数为4%的七氟醚+2 L·min^(−1)氧气),持续吸入6 h。用Morris水迷宫实验观察各组大鼠认知能力,用HE染色观察海马组织学形态,用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定海马组织中枢神经特异性蛋白(soluble protein 100β,S-100β)、神经元特异性烯醇化酶(neuron-specific enolase,NSE)、白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,流式细胞术测定神经细胞凋亡情况,蛋白印迹法(Western blotting)测定海马组织B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl2-associated X protein,Bax)、磷酸化磷脂酰肌醇3-激酶(phosphorylated phosphatidylinositol 3-kinase,p-PI3K)、磷酸化丝氨酸/苏氨酸激酶(phosphorylated serine/threonine kinase,p-Akt)和磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,p-mTOR)的水平。结果HE染色结果表明,对照组海马组织神经细胞结构正常,排列紧密,七氟醚各浓度组大鼠海马组织神经细胞减少,排列紊乱,分布不均匀。与对照组比较,七氟醚各浓度组大鼠逃避潜伏期时间,S100-β、NSE、IL-1β、TNF-α水平,Bax蛋白水平和神经细胞凋亡率均升高,穿越平台次数,平台停留时间,Bcl-2、p-PI3K、p-Akt和p-mTOR蛋白水平均降低(P<0.05);七氟醚各浓度组诸项指数水平变化规律相同,并呈剂量依赖性(P<0.05)。结论七氟醚麻醉可诱导大鼠神经细胞凋亡,使大鼠认知能力衰退,其可能与调控PI3K/Akt/mTOR信号通路有关。

齐墩果酸通过miR-18a-5p/STK4轴抑制白血病K562细胞恶性进展

慢性粒细胞白血病(chronic myeloid leukemia,CML)是由于骨髓造血干细胞的恶性增生导致的疾病。虽然酪氨酸激酶抑制剂(Tyrosine kinase inhibitors,TKI)对慢性粒细胞性白血病的治疗取得长足进展,但耐药问题仍未解决。因此,寻找新的治疗药物和靶点十分关键。有研究表明,miRNAs与慢性粒细胞白血病在内的多种肿瘤有密切联系,齐墩果酸(oleanolic acid,OA)对白血病细胞有较好的抑制效果。通过对转录物组测序结果发现,齐墩果酸可以显著上调丝氨酸/苏氨酸蛋白质激酶4(serine/threonine-protein kinase 4,STK 4)的水平,而miR1a-5p是STK 4的靶miRNA。因此,本文旨在探究齐墩果酸是否可以通过miR-18a-5p/STK4影响K562细胞恶性进展。K562细胞经齐墩果酸处理后差异表达基因富集在凋亡相关通路上。EdU实验和CCK-8实验结果发现,齐墩果酸降低K562细胞增殖能力(P<0.05)。qRT-PCR,免疫荧光和Western印迹结果显示,齐墩果酸处理后,K562细胞中STK 4蛋白质水平表达显著上调(P<0.05),miR-18a-5p表达下调(P<0.05)。在细胞中转染miR-18a-5p mimics,能显著抑制STK 4的表达(P<0.05);转染miR-18a-5p inhibitor能显著增加STK 4的表达(P<0.05)。活性氧(reactive oxygen species、ROS)检测和线粒体膜电位检测结果显示,齐墩果酸可以促进K562细胞中ROS的增加,降低线粒体膜电位。过表达STK 4后,与Vector组相比,细胞的线粒体膜电位下降;敲降STK 4可以逆转齐墩果酸组导致的线粒体膜电位下降。流式细胞术检测显示,齐墩果酸能明显促进细胞凋亡的发生,而转染miR-18a-5p mimics后凋亡率显著下调(P<0.05);过表达STK 4可以促进细胞从早期凋亡向晚期凋亡转化,而同时转染miR-18a-5p mimics可以抑制这一现象。我们的研究结果证实,齐墩果酸可以通过维持miR-18a-5p的低表达和保持STK 4的高表达状态来促进K562细胞的凋亡。

P38MAPK信号通路与uPA在卵巢癌细胞及组织中表达的相关性

背景与目的:P38丝裂原活化蛋白激酶(P38 mitogen-activated protein kinase,P38MAPK)信号通路参与多种肿瘤的发生、发展和转移过程,尿激酶型纤溶酶原激活剂(urokinase-type plasminogen activator,uPA)在肿瘤浸润和转移中发挥着重要作用。本实验研究卵巢癌组织中P38MAPK、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)、丝氨酸苏氨酸蛋白激酶(serine threonine kinase,AKT)及uPA的表达与临床病理特征的关系,并分析上述蛋白与u PA表达的相关性,探讨P38MAPK信号通路与uPA在卵巢癌细胞及组织中的表达及临床意义。方法:应用免疫组织化学法检测49例卵巢癌组织中u PA、P38MAPK、ERK和AKT蛋白的表达,采用蛋白[质]印迹法(Western blot)检测不同卵巢癌细胞系HO8910、HO-8910PM、SKOV3和CAOV3中uPA和P38MAPK蛋白的表达,使用特异性抑制剂SB203580阻断P38MAPK信号通路后检测u PA蛋白表达水平的变化。结果:uPA、P38MAPK、ERK和AKT蛋白在卵巢癌组织中的表达阳性率分别为61.22%、57.14%、53.06%和55.10%。uPA蛋白的表达与P38MAPK呈正相关(r=0.865,P=0.001),且与卵巢癌组织的临床病理分期(P=0.029)、分化(P=0.03)和转移程度(P淋巴=0.022,P大网膜=0.012)有关,而与患者的年龄(P=0.754)及组织学类型(P=0.652)无关。ERK、AKT蛋白的表达与卵巢癌淋巴结转移(PERK=0.011,PAKT=0.022)和大网膜转移(PERK=0.006,PAKT=0.000)有关,而与患者的年龄(PERK=0.000,PAKT=0.022)、组织类型(PERK=0.771,PAKT=0.245)及病理分期(PERK=1.000,PAKT=0.254)无关。卵巢癌细胞系HO-8910PM中uPA蛋白的表达水平明显高于HO8910、SKOV3和CAOV3细胞系,使用SB203580阻断P38MAPK信号通路后可降低uPA蛋白的表达,且随着SB203580浓度升高u PA蛋白表达水平逐渐降低。卵巢癌中P38MAPK及u PA蛋白的表达与卵巢癌的预后显著相关(Log-rank=3.897和11.044,P=0.048和0.001)。结论:卵巢癌组织中P38MAPK信号通路处于激活状态;P38MAPK信号通路的激活可上调u PA的表达,促进卵巢癌的恶性进展;P38MAPK信号通路和u PA可能在卵巢癌侵袭和转移的过程中发挥重要作用。P38MAPK和uPA蛋白有望成为卵巢癌预后评估的重要指标。

辛开苦降法对多囊卵巢综合征胰岛素抵抗患者卵巢功能、子宫内膜容受性及磷脂酰肌醇3激酶/丝苏氨酸蛋白激酶通路的影响

目的 观察辛开苦降法对多囊卵巢综合征(Polycystic ovarian syndrome,PCOS)胰岛素抵抗(Insulin resistance,IR)患者卵巢功能、子宫内膜容受性及磷脂酰肌醇3激酶/丝苏氨酸蛋白激酶(Phosphatidylinositol 3 kinase/serine threonine protein kinase,PI3K/AKT)通路的影响。方法 选取2018年11月—2020年1月期间四川中医药高等专科学校附属绵阳市中医医院收治的PCOS-IR患者83例,按随机数字表法分为对照组41例和治疗组42例。对照组采用炔雌醇环丙孕酮联合二甲双胍治疗,治疗组采用中医辛开苦降法(中药),均连续治疗12周。观察比较两组患者治疗前后体质量指数(Body mass index,BMI)、卵巢功能[卵泡刺激素(Follicle stimulating hormone,FSH)、黄体生成素(Luteinizing hormone,LH)、LH/FSH与抗缪勒管激素(Anti-mullerian hormone,AMH)]、血糖[空腹胰岛素(Fasting insulin,FIN)、空腹血糖(Fasting blood glucose,FPG)、餐后2 h血糖(2 hours postprandial blood glucose,2 h PBG)与胰岛素抵抗指数(Homeostasis model assessmentinsulin resistance,HOMA-IR)]、子宫内膜容受性[收缩期峰值流速(Vmax)、搏动指数(Pulse index,PI)、阻力指数(Resistance index,RI)、血流参数血管化指数(Vascularity index,VI)、血流指数(Flow index,FI)与血管化血流指数(Vascularity flow index,VFI)]、外周血PI3K/AKT通路(PI3K、AKT mNRA),治疗期间不良反应情况。结果 治疗后两组患者BMI、FPG、FIN、2 h GLU、HOMA-IR水平均较治疗前明显降低,差异有统计学意义(P<0.05);且治疗组BMI、FPG、FIN、2 h GLU、HOMA-IR水平均明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者AMH、LH、LH/FSH水平均较治疗前降低,差异有统计学意义(P<0.05);FSH较治疗前无显著变化,差异无统计意义(P>0.05);且治疗组AMH、LH、LH/FSH明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者子宫内膜厚度、子宫容积、Vmax、VI、FI、VFI均较治疗前明显升高,PI、RI均较治疗前降低,差异有统计学意义(P<0.05);且治疗组子宫内膜厚度、子宫容积、Vmax、VI、FI、VFI均明显高于对照组,PI、RI均明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者PI3K、AktmRN表达均较治疗前升高,差异有统计学意义(P<0.05);且治疗组PI3K、AktmRN表达均明显高于对照组,差异有统计学意义(P<0.05)。治疗期间,两组患者均未发生严重不良反应。结论 中医辛开苦降法不仅能改善PCOS-IR患者胰岛素抵抗,还可改善其卵巢功能、子宫内膜容受性,激活PI3K/AKT通路,安全性高。

FTO介导m6A修饰的PRKD2调节SIRT1/HIF-1α通路抑制糖尿病肾病足细胞损伤的机制研究

目的探究脂肪含量和肥胖相关蛋白(fat mass and obesity-associated protein,FTO)和丝氨酸-苏氨酸激酶蛋白激酶D2(serine-threonine kinase protein kinase D2,PRKD2)在糖尿病肾病(diabetic kidney disease,DKD)进展中的调控作用和调节机制。方法采用35 mmol/L葡萄糖对足细胞(MPC5细胞)进行高糖刺激24h构建DKD体外模型。采用FTO过表达载体(pcDNA-FTO)和PRKD2过表达载体(pcDNA-PRKD2),或空载体(vector)转染高糖诱导的MPC5细胞。通过RT-qPCR检测FTO和PRKD2过表达效率;MeRIP检测PRKD2 mRNA的N6-甲基腺苷(N6-methyladenosine,m6A)修饰水平;ELISA检测Caspase-3活性、IL-6,TNF-α和单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)分泌量;流式细胞术分析细胞凋亡率;Western blot评估FTO和PRKD2蛋白水平,以及SIRT1/HIF-1α通路关键蛋白表达水平;Pearson分析FTO和PRKD2水平的相关性。结果与无高糖诱导对照组比较,高糖诱导的足细胞中FTO蛋白(0.51±0.04 vs 1.00±0.03)和PRKD2蛋白(0.45±0.03 vs 1.01±0.04)水平显著下调,差异具有统计学意义(t=13.17,16.76,均P<0.001)。高糖诱导的足细胞中FTO蛋白水平和PRKD2蛋白水平呈正相关(r2=0.7051,P<0.001)。与vector组相比,pcDNA-FTO组PRKD2 mRNA的m6A水平(0.56±0.09 vs1.01±0.13)降低,PRKD2 mRNA水平(3.16±0.14 vs 1.03±0.02)显著升高,差异具有统计学意义(t=51.37,11.82,均P<0.001)。与control组(IL-6:512.76±61.85 pg/ml,TNF-α:28.17±2.83 pg/ml,MCP-1:157.31±17.69 pg/ml)和vector组(IL-6:498.41±87.51 pg/ml,TNF-α:26.35±5.47 pg/ml,MCP-1:165.52±16.87 pg/ml)比较,pcDNA-PRKD2组IL-6(301.86±21.85 pg/ml),TNF-α(11.06±4.12 pg/ml),MCP-1分泌量(81.45±9.03pg/ml)显著减少,差异具有统计学意义(F=7.51,10.47,61.97,均P<0.01)。与control组(Caspase-3:689.65±79.5U/L,细胞凋亡率:22.31%±2.69%)和vector组(Caspase-3:715.91±113.58 U/L,细胞凋亡率:21.07%±3.28%)比较,pcDNA-PRKD2组Caspase-3活性(437.64±104.76 U/L)和细胞凋亡率(8.41%±3.15%)下降,差异具有统计学意义(F=2.35,79.13,均P<0.01)。与control组(SIRT1:1.01±0.05,HIF-1α:1.03±0.07)和vector组(SIRT1:0.97±0.05,HIF-1α:1.02±0.03)相比,pcDNA-PRKD2组SIRT1蛋白(3.51±0.15)水平升高,HIF-1α蛋白(0.37±0.07)水平降低,差异具有统计学意义(F=31.54,8.31,均P<0.01)。结论FTO介导m6A修饰的PRKD2通过SIRT1/HIF-1α通路抑制高糖诱导的足细胞炎症反应和细胞凋亡。

BRAF、TP53、Pax8-PPARγ在甲状腺癌中的表达及疗效预测价值

目的探讨肿瘤蛋白P53(TP53)、丝氨酸/苏氨酸蛋白激酶(BRAF)、配对盒基因8-过氧化物酶体增殖物激活受体γ(Pax8-PPARγ)在甲状腺癌中的表达及疗效预测价值。方法将2020年4月至2022年4月该院收治的150例甲状腺癌患者纳入研究。检测患者甲状腺癌组织及癌旁组织中TP53、BRAF、Pax8-PPARγmRNA的表达水平,分析其与临床病理因素的关系,基于受试者工作特征(ROC)曲线及决策曲线分析TP53、BRAF、Pax8-PPARγmRNA表达水平预测^(131)I治疗效果的价值。结果甲状腺癌组织中的TP53、BRAF、Pax8-PPARγmRNA表达水平高于癌旁组织(P<0.05)。甲状腺癌患者淋巴结转移、肿瘤最大径、包膜浸润与TP53、BRAF、Pax8-PPARγmRNA表达水平有关(P<0.05)。采用放射性^(131)I清除术后残留的甲状腺组织(简称清甲)失败患者组织TP53、BRAF、Pax8-PPARγmRNA表达水平均高于清甲成功患者(P<0.05)。ROC曲线分析显示,TP53、BRAF、Pax8-PPARγmRNA联合预测甲状腺癌患者清甲失败的曲线下面积优于三者单独预测(P<0.05)。决策曲线显示,三者联合预测甲状腺癌清甲失败发生的净收益率优于单一预测(P<0.05)。结论TP53、BRAF、Pax8-PPARγ在甲状腺癌组织中呈高表达,联合检测有助于预测^(131)I治疗效果,为临床确定合理治疗方案及时机提供参考。

LRRK2基因Gly2385Arg多态性与泉州地区汉族人群散发性帕金森病的关联研究

目的探讨LRRK2基因Gly2385Arg同泉州地区汉族人群散发性帕金森病(PD)的关系。方法收集268例泉州地区汉族散发性PD患者和277例健康者的外周血液标本并提取DNA,利用聚合酶链反应(PCR)-限制性酶切方法(RFLP)进行LRRK2基因Gly2385Arg多态位点的基因型检测,变异者进行直接测序验证。结果 Gly2385Arg检测中,PD组有26例是杂合型变异(9.7%),显著高于对照组(2.2%;P<0.01,OR:4.85,95%CI:1.96～11.99)。按性别分层显示:在男性亚组或女性亚组中,Gly2385Arg变异的频率在PD组均分别显著高于健康对照组(男性亚组:P<0.01,OR:4.07,95%CI:1.32～12.54;女性亚组:P<0.01,OR:6.44,95%CI:1.39～29.72)。按起病年龄分层显示,晚发型PD(起病年龄>50岁)中,Gly2385Arg变异的频率显著高于对照组(P<0.01,OR:4.44,95%CI:1.63～12.06);早发型PD(起病年龄≤50岁)中,Gly2385Arg变异的频率与对照组差别无意义。结论 LRRK2基因Gly2385Arg多态是泉州地区汉族人群晚发型PD的一个风险因子。

磷酸化修饰介导的病原微生物致病机制研究

病原微生物通过特殊的分泌系统向宿主释放一系列效应蛋白,干扰宿主的先天免疫相关信号通路,引起疾病的发生甚至导致宿主死亡,是病原微生物致病的重要手段。翻译后修饰在病原微生物感染过程中发挥重要功能,受到越来越多重视,其中,磷酸化修饰是一种常见的翻译后修饰,迄今为止许多效应蛋白被发现具有丝氨酸/苏氨酸激酶活性。微生物借助于这些具有激酶活性的效应蛋白,将宿主内特异靶蛋白进行磷酸化修饰,进而干扰宿主细胞内的信号转导功能,抑制宿主免疫防御反应,促进病原微生物在宿主体内的繁殖与扩增。对近期发现的具有激酶活性的效应蛋白进行概述,为进一步理解磷酸化修饰介导的病原微生物感染以及药物靶标选择提供参考。

NEK2基因rs10494938、rs10158205位点多态性与肝细胞癌家系遗传易感性的关系

目的探讨丝/苏氨酸激酶NIMA相关激酶2(NEK2)基因rs10494938、rs10158205位点多态性与肝细胞癌(简称肝癌)家系遗传易感性的关系。方法选取广西扶绥县肝癌高发地区20个肝癌家系,包括肝癌患者20例及其直系亲属59例;另选10个正常家系(健康人群)进行对照,共40例。各家系均符合Hardy-Weinberg遗传平衡,均具有人群代表性。采用质谱方法检测NEK2基因rs10494938和rs10158205位点基因型分布频率;采用非条件Logistic回归分析法分析各家系人群等位基因及基因型分布频率的差异。结果 NEK2基因rs10494938位点存在CC和CG两种基因型,其中CC基因型114例、CG基因型5例;rs10158205位点存在CC、CT和TT三种基因型,其中CC基因型69例、CT基因型45例、TT基因型5例。肝癌家系中肝癌患者及其直系亲属、正常对照家系NEK2基因rs10494938位点CC、CG基因型及等位基因G、C分布频率比较差异均无统计学意义(P均>0.05),rs10158205位点CC、CT、TT基因型及等位基因T、C分布频率比较差异均无统计学意义(P均>0.05)。结论 NEK2基因rs10494938、rs10158205位点多态性与广西扶绥县肝癌家系遗传易感性之间无相关性。

TGFβ1-Smad/p38 MAPK信号通路与肾移植患者术后肾功能指标的相关性

目的:探讨转化生长因子β1(TGF-β1)/丝/苏氨酸激酶受体(smad)/p38丝裂原活化蛋白激酶(p38 MAPK)信号通路与肾移植患者术后肾功能指标的相关性。方法:选取420例肾移植手术患者为研究对象,根据术后效果分为稳定组(n=275)和肾纤维化组(n=145)。比较两组患者肾功能指标(Scr、BUN)水平、炎症因子指标(TNF-α、IL-6、IL-10)水平及TGF-β1、Smad7、p38 MAPK水平,分析TGFβ1-Smad/p38 MAPK信号通路与肾移植患者术后肾功能指标的相关性。结果:稳定组患者TNF-α、IL-6、IL-10、Scr、BUN、TGF-β1、p38MAPK水平低于肾纤维化组(P<0.05);Smad 7高于肾纤维化组(P<0.05)。相关性分析显示,TGF-β1、p38 MAPK、TNF-α、IL-6、IL-10水平与肾移植患者术后Scr、BUN呈正相关(P<0.05),Smad 7与Scr、BUN呈负相关(P<0.05)。结论:TGFβ1-Smad/p38 MAPK信号通路可通过触发炎症反应参与肾移植患者术后肾纤维化,导致肾移植患者术后肾功能变化。