As one of the basic processes of cellular activity,endocytosis sorting determines the fate of many proteins on the endosomal membranes,thus affecting a variety of important cellular processes,such as nutrient uptake,cell signal transduction,intracellular ion homeostasis,etc.Its dysregulation is closely related to various neurological diseases,metabolic disease,and immune disorders[1].Early endosomes(EEs)are crucial in cargo sorting within vesicular trafficking,cargos destined for the plasma membrane or the Golgi are first sorted into the specific membrane structure of the EE-EE buds,and then separated from the EE by membrane fission,and further returned to the plasma membrane or Golgi through a recycling pathway.Cargoes that are destined for degradation remain in EE,and as the endosome matures,they are ultimately degraded in the lysosome.Therefore,the EE buds play a key role in cargo sorting[2].Currently,the molecular mechanism of EE bud biogenesis is still not fully understood,and the molecular mechanism of membrane lipid regulation during budding is still unknown.

目的:研究VPS26在高脂环境中对大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSC）成骨、成脂分化作用的机制，探讨VPS26对高脂大鼠种植体骨结合和裸鼠异位成骨的影响。方法:普通成骨诱导液（成骨组）和高脂成骨诱导液（高脂组）培养BMSC，高脂组促进或抑制VPS26的表达，检测成骨和成脂相关基因的表达。细胞诱导第7、14天后行碱性磷酸酶（alkaline phosphatase，ALP）和油红O染色；成骨组细胞采用免疫荧光染色和免疫共沉淀方法检测VPS26与β-联蛋白（β-catenin）的结合，双荧光素酶报告实验检测萤火虫荧光值/海肾荧光值（以下简称TOP/FOP）比值。取18只雄性12周龄高脂血症Wista大鼠（体质量为160~200 g），于其双侧股骨干骺端植入种植体，分为3组，每组6只，分别注射VPS26过表达慢病毒（LV-VPS26组）、阴性对照慢病毒（LV-nc组）和生理盐水（空白对照组），种植体及周围骨组织行显微CT分析和HE、油红O染色评估种植体骨结合和股骨脂滴形成。20只雌性6周龄裸鼠（体质量为30~40 g）均分为5组，每组4只，于背部皮下分别植入不转染和转染了LV-VPS26、LV-nc、shVPS26、shscr慢病毒的成骨组BMSC，取样观察异位成骨。结果:过表达VPS26后高脂组BMSC中ALP的mRNA表达水平（1.56±0.09）显著高于阴性对照（1.01±0.03）（ t=10.09， P<0.001），过氧化物酶增殖物活化受体-γ（peroxisome proliferator-activated receptor-γ，PPAR-γ）和脂肪酸结合蛋白4（fatty acid-binding protein4，FABP4）的mRNA表达水平则显著低于阴性对照（ t=6.44， P<0.001； t=10.01， P<0.001）。蛋白质印迹法结果显示过表达VPS26后高脂组BMSC中ALP、Runt相关转录因子2的蛋白表达较阴性对照增强，PPAR-γ、FABP4则减弱，高脂组骨髓间充质干细胞ALP活性在过表达VPS26后更强，脂滴的形成较阴性对照更弱，数量更少。免疫荧光染色、免疫共沉淀和双荧光素酶报告实验结果显示VPS26与β-catenin存在共定位和相互作用，且TOP/FOP比值显著上升43.10%（ t=-3.17， P=0.034）。VPS26过表达后高脂大鼠种植体骨结合增强而脂滴数量下降，HE染色结果显示VPS26过表达后裸鼠皮下支架周围组织骨量增多。 结论:VPS26通过Wnt/β-catenin通路激活BMSC成骨分化且抑制成脂分化，具有促进高脂大鼠种植体骨结合和裸鼠异位成骨的作用。

Background::Acute coronary syndromes mainly result from abrupt thrombotic occlusion caused by atherosclerotic vulnerable plaques (VPs) that suddenly rupture or erosion. Fibrous cap thickness (FCT) is a major determinant of the propensity of a VP to rupture and is recognized as a key factor. The intensive use of statins is known to have the ability to increase FCT; however, there is a risk of additional adverse effects. However, lower dose statin with ezetimibe is known to be tolerable by patients. The present study aimed to investigate the effect of intensive statin vs. low-dose stain + ezetimibe therapy on FCT, as evaluated using optical coherence tomography. Method::Patients who had VPs (minimum FCT <65 μm and lipid core >90°) and deferred from intervention in our single center from January 2014 to December 2018 were included in the trial. They were divided into the following two groups: intensive statin group (rosuvastatin 15-20 mg or atorvastatin 30-40 mg) and combination therapy group (rosuvastatin 5-10 mg or atorvastatin 10-20 mg + ezetimibe 10 mg). At the 12-month follow-up, we compared the change in the FCT (ΔFCT%) between the two groups and analyzed the association of ΔFCT% with risk factors. Fisher exact test was used for all categorical variables. Student’s t test or Mann-Whitney U-test was used for analyzing the continuous data. The relationship between ΔFCT% and risk factors was analyzed using linear regression analysis. Result::Total 53 patients were finally enrolled, including 26 patients who were in the intensive statin group and 27 who were in the combination therapy group. At the 12-month follow-up, the serum levels of total cholesterol (TC), total triglyceride, low-density lipoprotein (LDL-C), hypersensitive C-reactive protein (hs-CRP), and lipoprotein-associated phospholipase A2 (Lp-PLA2) levels were reduced in both the groups. The ΔTC%, ΔLDL-C%, and ΔLp-PLA2% were decreased further in the combination therapy group. FCT was increased in both the groups (combination treatment group vs. intensive statin group: 128.89 ± 7.64 vs. 110.19 ± 7.00 μm, t = -9.282, P < 0.001) at the 12-month follow-up. The increase in ΔFCT% was more in the combination therapy group (123.46% ± 14.05% vs. 91.14% ± 11.68%, t = -9.085, P < 0.001). Based on the multivariate linear regression analysis, only the serum Lp-PLA2 at the 12-month follow-up ( B = -0.203, t = -2.701, P = 0.010), ΔTC% ( B = -0.573, t = -2.048, P = 0.046), and Δhs-CRP% ( B = -0.302, t = -2.963, P = 0.005) showed an independent association with ΔFCT%. Conclusions::Low-dose statin combined with ezetimibe therapy maybe provide a profound and significant increase in FCT as compared to intensive statin monotherapy. The reductions in Lp-PLA2, ΔTC%, and Δhs-CRP% are independently associated with an increase in FCT.

目的:探讨婴幼儿脑室出血后脑积水(posthemorrhagic hydrocephalus，PHH)的相关因素。方法:回顾性分析2013年6月至2022年1月郑州大学第三附属医院神经外科收治的112例脑室出血(intraventricular hemorrhage，IVH)患儿临床资料，通过随访IVH患儿是否发展为脑积水来探讨PHH的相关因素。将随访过程中出现脑积水者纳入PHH组，未出现脑积水纳入无脑室扩张(resolved ventricular dilation，RVD)组。采用单因素分析及多因素Logistic回归分析探讨IVH后发生脑积水的相关因素。结果:112例中，55例(55/112，49%)发生PHH；58例(51.79%)行单纯保守对症治疗，28例(25%)行腰椎穿刺，26例(23.21%)行VPS治疗。5例(4.46%)死亡，其中2例死于肺功能不全，3例死于多器官功能衰竭；107例存活患儿中，随访期间出现3例(2.80%)癫痫，1例(0.93%)视力障碍，6例(5.61%)听力障碍，5例(4.67%)脑性瘫痪。多因素Logistic回归分析结果显示：出生时胎龄28～32周( OR＝19.078，95% CI: 2.408～151.140， P＝0.005)、产前未使用类固醇( OR＝20.642，95% CI：1.882～226.404， P＝0.013)、脑室出血级别高( OR＝94.193，95% CI：16.583～535.043， P<0.001)是PHH发生的独立危险因素。 结论:IVH患儿低胎龄，脑室出血级别高，产前未使用类固醇均与婴幼儿PHH存在关联。围术期应关注IVH患儿胎龄、产前使用类固醇情况及脑室出血级别，及时干预，避免其发展为PHH，从而改善患儿预后。

目的:探讨脑室-腹腔分流术(ventriculo-peritoneal shunt，VPS)治疗艾滋病合并新型隐球菌性脑膜炎(cryptococcal neoformans meningitis，CNM)患者的临床疗效。方法:纳入2015年1月至2020年6月广州医科大学附属市八医院感染科住院治疗的艾滋病合并CNM患者，分为VPS组和常规治疗组。回顾性分析患者入院时脑膜炎相关症状和体征、脑脊液压力、脑脊液常规检查、脑脊液墨汁涂片染色、隐球菌培养结果，以及隐球菌培养转阴时间等资料，比较2组患者治疗6周和48周时的疗效。统计学分析采用两独立样本 t检验或 χ2检验。 结果:66例艾滋病合并CNM患者中，VPS组26例，年龄为(35.7±11.9)岁，11例(42.3%)入院时脑脊液压力>330 mmH 2O(1 mmH 2O＝0.009 8 kPa)，25例(96.2%)墨汁染色阳性，20例(76.9%)脑脊液新型隐球菌培养阳性；常规治疗组40例，年龄为(38.9±12.9)岁，15例(37.5%)脑脊液压力>330 mmH 2O，32例(80.0%)墨汁染色阳性，31例(77.5%)脑脊液新型隐球菌培养阳性，两组患者年龄、脑脊液压力>330 mmH 2O者所占比例、墨汁染色阳性率、脑脊液培养阳性率之间差异均无统计学意义( t＝-1.02, χ2＝0.15、3.49、0.00，均 P>0.050)。所有患者入院后均给予抗真菌治疗、降颅压、营养支持和对症治疗，VPS组内科保守治疗欠佳后行VPS。治疗6周时，VPS组脑脊液压力恢复正常率为57.7%(15/26)，部分缓解率为73.1%(19/26)，分别高于常规治疗组的31.0%(9/29)和47.5%(19/40)，差异均有统计学意义( χ2＝3.96、4.22，均 P<0.050)。治疗48周时，VPS组脑脊液压力恢复正常率为92.3%(24/26)，脑脊液隐球菌培养转阴率为100.0%(20/20)，完全缓解率为46.2%(12/26)，分别高于常规治疗组的37.9%(11/29)、67.7%(21/31)和20.0%(8/40)，差异均有统计学意义( χ2＝17.52、8.03、5.10，均 P<0.050)。VPS组完全或部分缓解者22例，无效4例，无一例死亡；常规治疗组完全或部分缓解者23例，无效12例，死亡5例，VPS组无效或死亡患者比例为15.4%(4/26)，低于常规治疗组的42.5%(17/40)，差异有统计学意义( χ2＝5.34， P＝0.021)。 结论:艾滋病合并CNM患者行VPS能够提高疗效，改善预后，降低治疗无效或病死率。

目的 探讨神经内镜下第三脑室底造瘘术(ETV)治疗梗阻性脑积水的长期随访效果.方法 回顾性分析2010年1月至2018年8月EVT治疗的142例梗阻性脑积水的临床资料.结果 142例均顺利完成手术,造瘘口大小2～10 mm,平均5 mm.术后CT显示脑积水均缓解,其中119例(83.8%)症状逐步改善,23例无明显改善.术后发生颅内出血26例(18.3%),保守治疗后恢复;发生颅内感染6例(4.2%),药物治疗后恢复;未见明显偏瘫、意识障碍、尿崩、明显记忆力障碍、基底动脉损伤、动眼神经损伤情况.失访10例,其余132例随访1～147个月,平均89.7个月,其中118例(89.4%)随访超过36个月.首次ETV失败10例(7.6%),发生时间在术后1～37个月,其中8例行CT检查后改行VPS,1例再次行ETV,1例术后5个月因急性脑积水行脑室外引流术后出现硬膜下血肿而死亡.结论 ETV是治疗梗阻性脑积水的首选方式,长期有效率高,失败多发生于术后3年内,注意随访,根据病情可再次ETV或VPS解决脑积水.

目的 探讨气腹下腹腔穿刺置管术在脑积水腹腔分流术中的应用价值.方法 回顾性分析2002年6月至2016年12月浙江省立同德医院收治的490例脑积水患者的临床资料,以2002年6月至2009年1月采用开腹置入分流管行脑室-腹腔分流术(VPS)的147例为对照组(开腹置管组),以2009年2月至2016年12月采用腹腔穿刺置入分流管行分流术的343例为观察组(腹腔穿刺置管组);再将腹腔穿刺置管组患者按置管方式不同分为VPS组228例和腰大池-腹腔分流术(LPS)组115例.收集开腹置管组和腹腔穿刺置管组患者术后6个月感染、分流管堵塞(堵管)、出血和其他并发症(如引流不足或过度、出血、癫痫、肠梗阻)发生情况等临床资料,比较各组并发症发生率的差异.结果 开腹置管组感染〔10.20%(15/147)比3.79% (13/343)〕、堵管〔14.29%(21/147)比7.58%(26/343)〕和其他并发症〔23.13%(34/147)比10.79%(37/343)〕发生率明显高于腹腔穿刺置管组(均P<0.05);VPS组堵管率明显高于LPS组〔9.65%(22/228)比3.48%(4/115), P<0.05〕.结论 气腹下腹腔穿刺置入分流管行脑积水分流术操作简便、安全,能有效降低感染率和堵管率;其中LPS堵管发生率比VPS更低.

目的 研究胃癌细胞在表皮生长因子(EGF)刺激下表皮生长因子受体(EGFR)相互作用蛋白的时序性变化.方法 筛选高表达EGFR的胃癌细胞系;EGF刺激不同时间后,利用甲醛固定免疫沉淀联合质谱(IP-XL/MS)技术对胃癌细胞中EGFR相互作用蛋白表达谱进行无标定量,捕获EGFR网络的动态变化.结果与结论 8种胃癌细胞系中,SGC7901细胞表达EGFR量最高. 4次生物学重复实验,共鉴定到3728个蛋白,625个EGFR相互作用蛋白,包括已知的相互作用蛋白( GRB2、CBL、SHC1 等) ,并发现59 个新蛋白( ANKFY、LGR4、SNX3、VPS26 A、ZFYVE20等). ANOVA检验分析得到406个响应EGF刺激的差异表达蛋白,根据动力学变化可分成5个簇,其具有不同的生物学功能,如蛋白转运、內吞、囊泡循环和蛋白酶体输运等,共同参与EGFR信号传递的动态调控,交叉调节多条信号通路.

目的 研究腰大池腹腔分流术(LPS)治疗创伤后脑积水(PTH)的临床疗效.方法 回顾性分析2016年1月—2018年12月四川大学华西医院神经外科收治的创伤后脑积水患者172例,男性116例,女性56例;年龄24～73岁,平均51.2岁.按照手术方式不同分为腰大池腹腔分流术组(LPS组,83例)与脑室腹腔分流组(VPS组,89例),LPS组男性57例,女性26例;年龄24～69岁,平均50.7岁,GCS评分(9.7±3.7)分,ISS评分(21.7±4.5)分,APACHE II评分(16.8±2.9)分,Evans指数(0.364±0.083)分.VPS组男性59例,女性30例;年龄29～73岁,平均51.6岁,GCS评分(9.8±2.4)分,ISS评分(22.7±4.2)分,A-PACHE II评分(15.8±3.6)分,Evans指数(0.345±0.076)分.对两组患者临床资料、临床疗效指标、并发症发生情况和预后指标进行统计学分析.结果 VPS组并发症19例(21.3％)多于LPS组7例(8.4％),差异有统计学意义(P<0.05).LPS组症状改善分数(3.820±1.092)分高于VPS组(2.960±0.793)分(P<0.01).VPS组脑室Evans指数(0.338±0.030)分高于LPS组(0.278±0.036)分(P<0.01).VPS组改良RANKIN量表(2.980±1.413)分高于LPS组(1.980±1.104)分(P<0.01).LPS组GOS评分(3.780±1.213)分高于VPS组(2.100±1.085)分(P<0.01).结论 LPS可能是治疗PTH的一种更为有效的手术方法,并发症较少,预后可能更好.

A myriad of abiotic stress responses in plants are controlled by abscisic acid(ABA)signaling.ABA recep-tors can be degraded by both the 26S proteasome pathway and vacuolar degradation pathway after pro-cessing via the endosomal sorting complex required for transport(ESCRT)proteins.Despite being essen-tial for ABA signaling,the upstream regulators of ESCRTs remain unknown.Here,we report that the ESCRT-I component VPS23A is an unstable protein that is degraded via the ubiquitin-proteasome system(UPS).The UEV domain of VPS23A physically interacts with the two PSAP motifs of XBAT35,an E3 ubiquitin ligase,and this interaction results in the deposition of K48 polyubiquitin chains on VPS23A,marking it for degradation by 26S proteasomes.We showed that XBAT35 in plants is a positive regulator of ABA re-sponses that acts via the VPS23A/PYL4 complex,specifically by accelerating VPS23A turnover and thereby increasing accumulation of the ABA receptor PYL4.This work deciphers how an ESCRT component is regulated in plants and deepens our understanding of plant stress responses by illustrating a mechanism whereby crosstalk between the UPS and endosome-vacuole-mediated degradation pathways controls ABA signaling.