Objective:To explore the potential target of unfolded protein response-protein kinase R-like endoplasmic reticulum kinase (UPR-PERK) pathway in the process of ER stress in liver, we also tested whether the UPR-PERK pathway could regulate the oxidative stress-induced growth inhibitor 1 ( Osgin1) in the liver endoplasmic reticulum (ER) stress models. Methods:Using GEO database as the source of analysis data, the differentially expressed genes were selected by GEO2R software, using Venn diagram, we identified Osgin1 as the gene with significant variation in the three physiological factors of fasting-refeeding, exercise, and age. The direct interaction between the UPR-PERK pathway and Osgin1 was verified using the GENEMANIA database. Subsequently, we used three different liver endoplasmic reticulum stress models (acute endoplasmic reticulum stress model, non-alcoholic fatty liver disease model and liver cancer model) in GEO Database and TCGA database to further verify the regulatory relationship of UPR-PERK pathway to Osgin1. GraphPadPrism 9.0.0 software was used for statistical analysis. Pearson correlation analysis was used to analyze the correlation between the transcriptional levels of UPR-PERK pathway genes and Osgin1. Results:Compared with the control group, fasting-refeeding, increased exercise and aging significantly changed the transcription level of Osgin1 ( Osgin1 mRNA was up-regulated by 700%, down-regulated by 156% and 229%, respectively, P<0.05). The GENEMANIA database verified the direct interaction between the UPR-PERK pathway and Osgin1. In the model of endoplasmic reticulum (ER) stress, compared with control group, inhibition of UPR-PERK pathway related gene ( Eif2ak3) could significantly reverse the up-regulation of Osgin1 mRNA induced by Tunicamycin ( Osgin1 mRNA was downregulated by 87%, P<0.001). In the non-alcohol fatty liver model, compared with control group, when the UPR-PERK pathway was alleviated by the administration of drugs, the Osgin1 transcription level was also down-regulated by 48%, 43.3% and 56.6% with BI4556906, EX10970 and IXA4 interventions ( P<0.05). In hepatic cancer cell tissues, compared with control group, the transcription levels of UPR-PERK pathway-related genes and Osgin1 were significantly up-regulated 109% ( P<0.001). Conclusions:The activation of UPR-PERK pathway upregulates Osgin1 transcription in physiological conditions and liver endoplasmic reticulum stress models.

目的:了解宁波某企业体检人群中甲状腺结节检出率,并分析相关影响因素.方法:收集宁波某大型国有企业5416名体检人员的体检资料.将人群分为甲状腺结节组与非甲状腺结节组,组间比较根据数据类型分别用两独立样本t检验、χ2检验进行分析,并用多因素Logistic回归分析受检人群中甲状腺结节检出率的影响因素.结果:甲状腺结节总检出率为35.8％,其中女性为48.4％,男性为32.6％;不同年龄段女性检出率均高于男性,且甲状腺结节检出率随年龄增长而显著升高(P<0.05).多因素Logistic回归分析显示,性别、年龄和腰围是甲状腺结节的可能影响因素(P<0.05).结论:宁波某企业体检人群中女性甲状腺结节检出率明显高于男性,性别、年龄和腰围与甲状腺结节有显著相关性.

线粒体脑肌病伴高乳酸血症和卒中样发作(MELAS)是母系遗传性线粒体疾病,临床表现多样,易与单纯疱疹病毒性脑炎(HSE)混淆.该文报道1例初诊时误诊为HSE的MELAS患者,该例患者因反复发热、头痛、肢体抽搐1个月,再发头痛1周就诊,入院时初步疑诊为HSE,予以抗病毒治疗无效,进一步行血液和尿液基因检测确诊为MELAS.MELAS可与不典型的HSE表现相似,应谨慎鉴别.脑脊液和(或)血清乳酸升高和基底节钙化有助于诊断MELAS,MELAS的线粒体DNA突变可通过血液和尿液基因检测,而不需要采用肌肉活组织检查这样的有创检查.

目的 探讨直肠腔内超声(ERUS)显示Denonvilliers筋膜解剖结构特点的可行性.方法 回顾性分析15例男性直肠癌患者术前ERUS图像,观察辨别Denonvilliers筋膜解剖结构特点,并与术中腹腔镜所见进行对照.其中1例实施术中ERUS与腹腔镜下所见进行对照以验证术前ERUS识别的结构是否为Denonvilliers筋膜.结果 在ERUS图像上,走行于直肠固有筋膜与前列腺、精囊腺之间的线状高回声部位为Denonvilliers筋膜,显示率达100％.15例术前ERUS图像与术中腹腔镜所见均相符.Denonvilliers筋膜与前列腺后方包膜贴合紧密,在精囊后方有疏松较宽间隙.在精囊腺后方及侧方存在结构个体差异,依据线状高回声数量的不同可分为单层结构(3例)及多层结构(12例).结论 ERUS可以准确显示Denonvilliers筋膜解剖结构特点.

目的 总结75例疑似重症肌无力患者行新斯的明试验的护理经验.方法 护理人员用药前对患者进行心理护理、用药指导并评估患者病情、测量生命体征,用药后监测患者生命体征、观察重症肌无力症状改善情况及药物副作用,观察有无严重不良反应发生.结果 ?75例患者均顺利完成试验.结论 通过观察患者无严重不良反应发生,用药副作用均能及时发现并对症处理.新斯的明试验是诊断重症肌无力的简单有效安全的方法 ,可以安全迅速判断患者是否为重症肌无力.

中枢神经系统（CNS）诺卡菌感染在临床上罕见且诊断较困难。该文报道了3例经基于宏基因组学的二代基因测序技术（mNGS）诊断的CNS诺卡菌感染病例。1例头颅MRI示脑膜脑炎,1例头颅MRI示右侧额顶枕叶、左侧大脑脚脑桥及小脑半球、脑膜多发病灶,1例头颅MRI示脑膜脑炎并双侧额叶、左侧侧脑室后角多发脓肿。3例脑脊液涂片均未发现病原体,2例脑脊液培养阴性。最后3例均经脑脊液mNGS被确诊为诺卡菌脑膜脑炎,1例予磺胺甲口恶唑、米诺环素口服,联合利奈唑胺静脉滴注抗感染后好转出院,随访无复发;1例予磺胺甲口恶唑口服,美罗培南、利奈唑胺静脉滴注等治疗,曾出现急性脑积水,先后行侧脑室外引流术、脑室腹腔分流术,其后病情逐渐好转并出院,随访无复发;1例予磺胺甲口恶唑口服,利奈唑胺、美罗培南静脉滴注,曾出现脑积水,行侧脑室外引流术,其后病情好转并出院,随后失访。该文提示在传统检测方法的基础上,将脑脊液mNGS作为常规的检测手段有助于提高该病的确诊率。

目的 探究伴发病对隐球菌性脑膜炎患者临床治疗效果的影响.方法 采用回顾性病例对照研究方法将327例非HIV非器官移植的患者基于入院时是否存在伴发病分为研究组(隐脑十伴发病172例)和对照组(隐脑无伴发病155例),并对两组患者的基本临床资料(性别、年龄、入院后首次脑脊液特征等)和临床治疗效果进行比较,分析伴发病的数量及种类对隐脑十伴发病组患者临床治疗效果的影响.结果 研究组脑脊液白细胞计数、蛋白、糖和氯化物含量与对照组比较差异均有统计学意义(P＜0.05);在进行10周的抗真菌治疗后,对照组患者治疗疗效优于研究组,差异有统计学意义(Z=-2.131,P=0.033);研究组中临床治疗成功与治疗失败患者在伴发病数量上的差异没有统计学意义;伴有消化系统和内分泌系统疾病的患者,治疗失败的人数更多,差异有统计学意义(P＜0.05).结论 入院时有伴发病的隐球菌性脑膜炎患者的治疗失败率更高,不良的预后结果与入院时伴发病数量无关,而与合并消化系统和内分泌系统疾病有关.

目的:探讨和分析早期康复护理用于重型颅脑损伤术后偏瘫患者的康复效果.方法:选取2016年10月至2017年9月期间,我院收治的82例重型颅脑损伤术后偏瘫患者作为研究对象;将全部82例患者以随机的方式分成各41例的两组,分别为观察组与对照组;对照组患者接受常规护理干预,观察组患者在此基础上接受早期康复护理干预;观察并对比两组患者的康复效果与术后并发症状况.结果:观察组患者术后生活质量评分改善状况优于对照组患者;同时,观察组患者的术后并发症发生率显著低于对照组患者,差异均具有统计学意义,P<0.05.结论:早期康复护理能够有效提高重型颅脑损伤术后偏瘫患者的康复效果,并能显著控制并发症状况,应当予以临床推广.

目的:探究急性心肌梗塞患者应用主动脉球囊进行治疗的临床效果与特征.方法:本次研究的分析对象是我院从2016年10月到2017年7月接收并治疗的89例急性心肌梗塞患者,且通过纳入标准均判定为急性心肌梗塞患者且并发心源性休克,并对抽取的急性心肌梗塞并心源性休克患者应用主动脉球囊反搏术进行治疗,探究并分析抽取的急性心肌梗塞患者采用主动脉球囊反搏术进行治疗的效果与特点,包括患者应用主动脉球囊治疗前后血液动力学的指标变化等.结果:抽取的89例心肌梗塞合并心源性休克患者应用主动脉球囊反搏术治疗后,结果显示,心肌梗塞患者的心指数3.99±1.05 L/min m2、心率83±12次/min、舒张压62.5±10.3mmHg、收缩压114.5±17.4mmHg明显优于治疗前,且通过主动脉球囊治疗后两小时患者的末梢循环良好,血压升高稳定,尿量增多且组织灌注增加.结论:针对心肌梗塞患者应用主动脉球囊进行治疗可以起到较好的效果,对于稳定患者血压,改善血流动力学指标价值较高,值得在临床治疗中应用.

Objective To investigate the diagnostic value of ultrasonography in the assessment of peri-pancreatic vascular invasion and resectability of pancreatic cancer. Methods Clinical data of 33 patients with pancreatic cancer admitted to the Third Affiliated Hospital of Sun Yat-sen University from January 2012 to September 2017 were retrospectively analyzed. The informed consents of all patients were obtained and the local ethical committee approval was received. Among them, 21 patients were male and 12 were female, aged 46-75 years with a median age of 54 years. Peri-pancreatic vascular invasion was evaluated by preoperative ultrasonography and the resectability of tumors was assessed. The ultrasonography findings were compared with the intraoperative exploration and surgical outcomes. And its sensitivity and specificity were calculated. Results The sensitivity, specificity, positive and negative predictive rates of ultrasonography in evaluating the peri-pancreatic vascular invasion were 88% (30/34), 99% (261/263), 94% (30/32) and 98% (261/265), respectively. For 19 patients with vascular invasion but without distant metastasis or lymph node metastasis, the sensitivity, specificity, positive and negative predictive rates of ultrasonography in assessing the unresectability were 5/6, 12/13, 5/6 and 12/13, respectively. Conclusions Ultrasonography can accurately and reliably evaluate the peri-pancreatic vascular invasion and give reference for assessing the resectability of pancreatic cancer. Key words: Ultrasonography; Pancreatic neoplasms; Neoplasm invasiveness; Diagnosis

Objective To investigate the antibacterial activity to Streptococcus mutans of a nisin⁃containing single⁃ bond universal adhesive. Methods Nisin was mixed into the bonding agent to produce concentrations ranging from 0.01 g/mL to 0.05 g/mL for the experiments, and adhesive without nisin was used as the control. Dentin⁃resin specimens were prepared for the microtensile strength test to evaluate changes in the bonding strength. The proper concentrations were selected for more tests. ① An agar diffusion test was applied with filter paper to detect the release of nisin, and adhesive without nisin was used as the negative control, 0.01 g/mL Nisin aqueous solution was used as the positive control. ② Solidification; resin adhesive specimens were prepared for the assessment of direct contact inhibition activity. ③ Confocal laser scanning microscopy was used to examine the effect of the adhesive on the biological film activity and the ability of Streptococcus mutans to produce extracellular polysaccharides. Results Nisin did not significantly reduce the bond strength of the modified adhesive at 0.01⁃0.03 g/mL (P < 0.05); these concentrations were selected for the subsequent antibiosis experiment. Rings could not be observed in the agar diffusion test, except for in the group of adhesive modified with 0.01⁃0.03 g/mL nisin. Resin adhesive with 0.01⁃0.03 g/mL nisin could significantly inhibit the proliferation of Streptococcus mutans on the surface of the specimens. The confocal laser scanning microscopy results indicate that only the adhesive resin modified with nisin could reduce the bacteria in the biofilm and the production of extracellular polysaccharides. Conclusion Single⁃bond universal adhesive with 0.01⁃0.03 g/mL nisin can inhibit the growth of Streptococcus mutans and its biofilms on the bonding interface, as well as decrease the production of extracellular polysaccharides, and thus has the potential to decrease the occurrence of secondary caries.

Objective To explore the clinical and head magnetic resonance imaging(MRI)charac-teristics of HIV-negative patients diagnosed with cryptococcal meningoencephalitis(CM). Methods Medical history,poultry exposure,clinical data,cerebrospinal fluid and head MRI findings of 126 non-AIDS CM pa-tients were collected. Based on the primary diseases,all patients were divided into two groups with or without predisposing factors and retrospectively analyzed. Results The main manifestations of CM were fever,head-ache,nausea,vomiting,and positive meningeal irritation,with increasing cerebrospinal fluid pressure and white blood cell count,and decreasing cerebrospinal fluid glucose. The majority of patients were apparently healthy (71 patient,56. 3%)and predisposing factors were idenfied in only 51 (43. 7%)patients,with hep-atitis B accounting for the most common underlying factor in these cases (28 patents,50. 9%). Poultry expo-sure history rate of 19. 7%in the group without prediposing factors,was significantly higher than which of 4%in the group with prediposing factors (P<0. 05). Meningeal irritation postive rate of 56. 4% and the average value of CSF glucose 2. 1 mmol/L in the group with predisposing factors was significantly higher than meningeal irritation postive rate of 43. 7% and the average value of CSF glucose 1 . 61 mmol/L (both P<0. 05 )in the group without predisposing factors. Whereas MRI findings manifested as meningeal enhancement and cerebral parenchymal lesions did not differ between two groups. Conclusions The final diagnosis of CM depends on pathological examination. CM mainly manifested as meningeal irritation and intracranial hypertension in clini-cal,and meningeal enhancement and cerebral parenchymal lesions in MRI findings,with certain characters,which is important for early diagnosis and differential diagnosis of CM.

Objective To investigate the regulating effect of dendritic cells (DC)over-expressing B-and T-lymphocyte attenuator (BTLA)upon the immune tolerance of mouse models with experimental autoim-mune encephalomyelitis (EAE).Methods The recombinant lentiviral vector carrying the target gene of BTLA was constructed.Bone marrow-derived DCs were isolated from the C57BL/6 mice,cultured and transfected at a multiplicity of infection (MOI)of 1 5 to construct DCs over-expressing BTLA.The C57BL/6 mouse models with EAE model were established.During the peak of onset,the spleen cells were isolated,and CD4 + T lym-phocytes were separated by magnetic beads and co-cultured with DCs over-expressing BTLA.The Th1 -type cy-tokine IFN-γand Th2-type cytokine IL-1 0 were quantitatively detected by ELISA.Results The recombinant LPP-Mm26645-Lv203-400 lentiviral vector carrying mouse BTLA gene was successfully constructed.DCs were transfected and cultured for 5 d at a MOI of 1 5.DCs over-expressing BTLA were detected after 96 h.The peak of onset was observed in C57BL/6 mice at 21 -24 d after MOG35-55 peptide immunization.CD4 +T lymphocytes and DCs over-expressing BTLA were co-cultured.ELISA demonstrated that the expression level of IFN-γwas significantly down-regulated,whereas the expression level of IL-1 0 was considerably up-regulated in the co-cul-ture system (both P <0.05).Conclusion Over-expressing of BTLA can enhance the tolerance of DCs,yield specific antigen immune tolerance to EAE via reversing Th1 /Th2 imbalance and negatively regulate the immune response of EAE.

Objective To evaluate the effect of different concentrations of glycogen synthase kinase-3(GSK3) inhibitor CHIR99021 upon regulating the differentiation of human induced pluripotent stem cells (iPSCs) into motor neurons, aiming to investigate the optimal concentration.Methods During the early stage of differentiation, iPSCs were co-cultured with different concentrations of CHIR99021 for 6 days and were divided into the control (0 μmol/L), 1, 2, 3 and 4 μmol/L groups.Immunofluorescence identification was observed under light microscope during the early, middle and advanced stages of induction and differentiation.Results Throughout the early, middle and advanced stages of differentiation, cell counting revealed that the cell quantity was increased along with the elevated concentration of CHIR99021.During the early stage, the expression level of nerve stem cell marker SOX1 did not significantly differ among 0, 1, 2 and 3 μmol/L groups.The expression of SOX1 in the 4 μmol/L group was significantly lower than that in the other groups (all P<0.05).In the middle stage, the expression level of motor neuron precursor marker Olig2 was significantly increased along with the elevated concentration of CHIR99021 in 0, 1, 2 and 3 μmol/L groups (all P<0.05).The expression of Olig2 in 4 μmol/L group was slightly lower than that in 3 μmol/L group(P<0.05).During the advanced stage, the cells in 2, 3 and 4 μmol/L groups expressed mature motor neuron marker choline acetyl transferase (ChAT) rather than in control and 1 μmol/L groups.Conclusions During the early stage of nerve differentiation, a proper concentration of CHIR99021 can promote the proliferation of iPSCs, enhance the efficiency of cell differentiation and eventually induce iPSCs into motor neurons.The optimal concentration of CHIR99021 is 3 μmol/L.

目的 比较超声造影(CEUS)与增强CT (CECT)、增强MRI (CEMRI)检查肝脏局灶性病变(FLLs)的安全性及耐受性.方法 收集2015年7月在中山大学附属第三医院同时行肝脏CEUS和CECT或CEMRI的肝病患者42例,其中37例同时接受CEUS和CECT检查,36例同时接受CEUS和CEMRI检查.观察检查结束后30 min内有否造影剂不良反应,并采用视觉模拟评分法(VAS)评估检查过程中的不适.结果 CEUS和CEMRI的造影剂不良反应发生率低于CECT(0比11％).患者做CEUS的VAS评分均低于做CECT或做CEMRI的评分(均为P＜0.05).结论 肝脏CEUS安全性高且耐受性好,是检查FLLs的首选方法.

Objective To construct lentiviral vector expressing B and T lymphocyte attenuator (BT-LA)gene and evaluate the feasibility of transfecting immature dendritic cells by this vector.Methods Ac-cording to the mRNA sequence of mouse BTLA published by GeneBank,the ORF lentiviral plasmid was con-structed and the recombinant lentiviral vector expressing BTLA was synthesized.Immature dendritic cells were subjected to primary isolation and culture from C57BL/6 mouse bone marrow.Immature dendritic cells were transfected with recombinant lentivial vector expressing BTLA at a multiplicity of infection of 1 5.The expres-sion level of BTLA was assessed by immunofluorescence and RT-PCR.Results The lentiviral vector LPP-Mm26645-Lv203-400 was successfully constructed at a titer of 3.25 ×1 09 TU /ml.At 96 h after immature den-dritic cells transfected by the lentiviral vector LPP-Mm26645-Lv203-400,positive cells expressing green fluo-rescent protein were observed under inverted fluorescent microscope.The expression level of BTLA in the trans-fected immature dendritic cells was significantly higher compared with that in the non-transfected cells (P <0.05),indicating the successful transfection.Conclusion The lentiviral vector expressing BTLA gene can be successfully transfected into the mouse marrow-derived immature dendritic cells.

目的 探讨用急性肾损伤网络(acute kidney injury network,AKIN)标准诊断心脏术后AKI的危险因素.方法 回顾性收集昆明医科大学第一附属医院心外科2012年6月至2013年6月期间所有行心脏手术的住院患者资料,根据AKIN诊断标准,对心脏术后AKI患者的发病率及危险因素进行统计学分析.结果 548例心脏手术患者中,用AKIN标准诊断心脏术后AKI的发生,其中81例发生AKI,发病率为14.78％,死亡患者3例;统计学分析表明:年龄、左室射血分数、高血压、术中甘露醇使用量、CPB时间、主动脉阻断时间、心脏停跳时间、机械通气时间、监护室停留时间、术前Scr、术前尿酸是心脏术后AKI发生的相关危险因素,而术前血肌酐(P＜0.01)、年龄(P＜0.01)、CPB时间(P＜0.01)、机械通气时间(P=0.026)是心脏术后AKI发生的独立危险因素.结论 AKI是心脏手术后严重的并发症,术前血肌酐、年龄、CPB时间、机械通气时间是心脏术后AKI的独立危险因素.

目的 探讨单层细胞分化法结合Smad信号双抑制剂,诱导人诱导多能干细胞(hiPSCs)分化为神经干细胞,并进一步分化为运动神经元的可行性.方法 无血清E8培养基培养hiPSCs,当其接近70％融合度时改用神经诱导培养基,并加入SB431542和DMH1.9d后将贴壁细胞消化,在含bFGF和EGF的培养基中悬浮培养,免疫荧光检测神经干细胞标记物的表达.在神经干细胞培养液中添加视黄酸(RA)和音猬因子(SHH),免疫荧光检测各分化阶段运动神经元相关标记物的表达.结果 经9d神经诱导,hiPSCs分化为表达Nestin、Sox1和Sox2的神经干细胞.神经干细胞在RA和SHH等的作用下进一步分化为运动神经元,免疫荧光检出各不同阶段标志物的表达.结论 单层分化法结合Smad信号双抑制剂,可将hiPSCs诱导为神经干细胞,并在RA和SHH等作用下进一步分化为运动神经元.

目的 探讨早期诊断心脏术后急性肾损伤(acute kidney injury,AKI)的标准,比较急性肾损伤网(acute kidney injury network,AKIN)及RIFLE标准对心脏术后AKI的发病率及预后差异.方法 收集昆明医科大学第一附属医院心外科2012年6月至2013年6月期间所有行心脏手术的住院患者资料,根据AKIN与RIFLE诊断标准,对心脏术后AKI患者的发病率进行统计学分析,比较AKIN及RIFLE标准对心脏术后AKI的发病率差异,并对心脏术后AKI患者进行电话随访调查,观察比较患者长期预后情况.结果 (1) 548例心脏手术患者中,根据RIFLE标准诊断心脏术后AKI的发生,与AKIN标准比较,发现AKIN标准诊断的1期AKI的发生率与相对应的RIFLE标准诊断的Risk期AKI的发生率明显升高,而2期AKI的发生率与Injury期AKI的发生率明显下降(P＜0.05),对于两者诊断的Ⅲ期AKI发生率无明显差别,两者差异无统计学意义(P＞0.05); (2)对于2诊断标准在AKI患者预后比较发现,AKIN标准对患者预后没有表现出优势.在对AKI患者进行电话随访调查中,53例参与随访,28例出现失访,失访率为34.57％;随访患者第1月、第3月、第6月肾功能与出院时肾功能比较,差异无统计学意义(P＞0.05).结论 (1) AKIN标准较RIFLE标准诊断术后AKI发病率稍高,但差异没有达到统计学意义; (2) AKIN与RIFLE两标准比较,对改善心脏术后AKI患者的预后无明显差异; (3) AKI患者出院后肾功能与出院时肾功能比较无明显差异.

Objective To explore the expression of Nogo-A and NgR in Ambystoma mexicanum brain tissue and the relationship between Nogo-A, NgR, and regeneration after central nervous system damage injury. Method The normal Ambystoma mexicanum brain tissue and the brain at 3, 7, 14, and 21 d after midbrain cholinergic neurons injury were collected to determine Nogo-A and NgR expression using immunofluorescence, western blot analysis, and Real-time Quantitative PCR(RT-PCR). Results Nogo-A and NgR had abundant expression in the normal ambystoma mexicanum brain. Three days after AF64A injection, we observed the significant loss of ChAT+ cells in the ambystoma mexicanum mesencephalic regions, as compared with the controls(P0.01). At 7, 14 and 21 days, ChAT+BrdU+ cells were observed in the lesioned animals and increased with time course. Nogo-A in the brain tissue had no significant changes after midbrain cholinergic neurons injury in mRNA and protein expression. However, a marked decrease of NgR expression both in mRNA and protein levels was found after midbrain cholinergic neurons injury compared with normal controls. Conclusion The study also demonstrated that the ability of regeneration in Ambystoma mexicanum central nervous system might correlate with the decreases of expression of the NgR after injury.