目的 探讨钙敏感受体(CaSR)拮抗剂溶钙素对骨密度和骨代谢的影响,为临床骨质疏松症的治疗提供新的思路.方法 C57BL6雌性小鼠36只,随机分为对照组(A组)、高钙高维生素D饮食组(B组)、溶钙素20 mg/kg灌胃治疗组(C组)、溶钙素50 mg/kg灌胃治疗组(D组)、特立帕肽(PTH(1-34)) 20 μg/kg皮下注射组(E组),小鼠分别处理12周后,测定各组小鼠海绵骨、皮质骨、全骨骨密度及骨组织中成骨关键基因如骨钙素(Bglap2)、Runt相关转录因子(Runx2) mRNA的表达,并探讨溶钙素治疗对小鼠血、尿钙的影响.结果 干预12周后,B、C、D、E组全骨骨密度与A组比较明显升高,差异均有显著性(F=39.82,P＜0.05);C、D组全骨骨密度水平较B组明显升高,差异有显著性(P＜0.05);而D组全骨骨密度与E组比较,差异无显著意义(P＞0.05).溶钙素可上调Bglap2、Runx2 mRNA的表达,改善骨转换,C、D、E组Bglap2、Runx2 mRNA表达量均较A组显著上调,差异有显著性(F=27.96、21.75,P＜0.05).此外,与钙和维生素D补充和外源性PTH治疗干预组相比,溶钙素对血钙波动的影响较小,差异无显著性(P＞0.05).溶钙素也可减少尿钙排泄,D组尿钙水平较B、E组显著降低(F=2.68,P＜0.05).结论 溶钙素可促进骨形成、增加骨密度,有可能成为骨质疏松症有效的治疗药物.

目的 探讨神经纤毛蛋白2单克隆抗体(NRP2mAb)对乳腺癌细胞增殖、迁移、侵袭及黏附作用的影响.方法 选取人乳腺癌细胞株MDA-MB-231,采用不同浓度NRP2mAb处理MDA-MB-231细胞后,采用CCK-8法检测细胞的增殖抑制率,Transwell小室法检测细胞的迁移和侵袭情况,纤连蛋白(FN)细胞黏附实验检测细胞的黏附能力.结果 NRP2mAb处理后,MDA-MB-231细胞的形态不规则,存在皱缩、脱落改变,细胞数量减少,细胞质颗粒数量增多.不同浓度NRP2mAb(25、50、75、100、200μg/ml)处理24、48、72 h后,MDA-MB-231细胞的增殖抑制率比较,差异均有统计学意义(P﹤0.05);且随着NRP2mAb浓度的增高,MDA-MB-231细胞的增殖抑制率逐渐增高;随着NRP2mAb作用时间的延长,NRP2mAb对乳腺癌细胞的抑制率逐渐增高.不同浓度NRP2mAb(0、25、50、75μg/ml)处理后,MDA-MB-231细胞的迁移数目、侵袭数目比较,差异均有统计学意义(P﹤0.01);且随着NRP2mAb浓度增高,MDA-MB-231细胞的迁移数目和侵袭数目均逐渐减少.与无FN组比较,0μg/ml NRP2mAb处理的FN组MDA-MB-231细胞的黏附率增高(P﹤0.05);FN组中,随着NRP2mAb浓度(0、25、50、75μg/ml)的增加,MDA-MB-231细胞的黏附率逐渐降低,组间两两比较,差异均有统计学意义(P﹤0.05).结论 NRP2mAb对乳腺癌细胞增殖、迁移、侵袭及黏附能力均有抑制作用,并具有浓度依赖性.

目的 桔梗皂苷D(platycodin D,PD)是中药材桔梗的主要活性成分,对多种肿瘤细胞具有抗肿瘤活性.有研究发现,它能够促进肝癌细胞的凋亡和抑制增殖,但对细胞侵袭性的影响在很大程度上是未知的.本实验研究PD对人肝癌HCCLM3细胞迁移侵袭作用的影响,并初步探讨其可能的作用机制.方法 不同浓度的PD处理肝癌HCCLM3细胞24和48 h,MTT法检测对细胞增殖的影响;划痕实验和Transwell迁移侵袭实验检测对细胞迁移和侵袭的作用;荧光定量PCR和蛋白质印迹实验分别检测基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和MMP-9基因在mR-NA和蛋白水平的表达情况.结果 MTT检测结果表明,PD降低肝癌HCCLM3细胞的存活率,抑制细胞的增殖,并呈剂量依赖性,P<0.01.划痕实验结果表明,10、20和40μmol/L的PD处理细胞48 h后,细胞的迁移率分别为(41.33±8.25)％、(31.87±9.83)％ 和(12.29±3.10)％,与对照组(86.53±6.04)％ 相比,差异有统计学意义,F=56.28,P<0.001.Transwell实验结果表明,10、20和40μmol/L的PD处理细胞48 h后,迁移的细胞个数分别为466.33±103.94、294.67±42.53和156.68±52.27,与对照组818.33±108.15相比,差异有统计学意义,F=36.47,P<0.001;侵袭的细胞个数分别为299.33±92.93、192.67±55.63和64.00±28.90,与对照组665.67±76.84相比,差异有统计学意义,F=43.48,P<0.001.荧光定量PCR和蛋白质印迹实验进一步研究表明,10、20和40μmol/L的PD处理细胞后,MMP-2和MMP-9在mRNA水平的表达降低,与对照组相比,差异有统计学意义,F值分别为16.86和27.32,P值分别为0.002和0.001;同时,MMP-2和MMP-9在蛋白水平的表达也降低,F值分别为8.85和15.01,P值分别为0.006和0.001.结论 PD能够抑制人肝癌HCCLM3细胞的迁移和侵袭能力,可能与降低MMP-2和MMP-9的表达有关.

Objective In recent years,the incidence of papillary thyroid carcinoma(PTC)increased gradually and the recurrence and metastasis of PTC patients are also increasing.The aim of this study was to investigate the effect of BRAFV600E mutation on the expression of HMGB1 in PTC,to explore the mechanism of BRAF gene affecting the development and prognosis of PTC,and then guiding clinical practice of precise therapy.Methods The preoperative serum and postoperative fresh tissue were collected from 44 cases of PTC patients in our hospital from September,2015 to December,2015.According to BRAFV600E mutations divided into positive group and negative group.Western blot,immunohistochemistry and fluorescence quantitative PCR were used to detect the level of HMGB1 in cancer tissues.The level of HMGB1 in serum was tested by ELISA method.Western blot was performed using Image J software.The relative quantitative method was used to calculate the fluorescence quantitative PCR data.The concentration of HMGB1 protein in serum was calculated by ELISA Calc regression/fit calculation program.All the data were analyzed by SPSS 20.0.Western blot,fluorescence quantitative PCR and ELISA data were analyzed by Chi-square test,Mann-Whitney-Wilcoxon test and independent sample t test.The relationship between lymph node metastasis and extrathyroid infiltration and the occurrence of BRAFV600E mutation was analyzed by chi-square test.Results In PTC cancer tissues,HMGB1 was mainly located in the cytoplasm,the levels of HMGB1mRNA and protein in positive group were lower than that of the negative group(Z =2.117,P ＜0.01,x2 =19.989,P ＜ 0.05),and the change did not show in peripheral blood(t =1.135,P ＞ 0.05).The expression of HMGB1 mRNA and protein in PTC with lymph node metastasis was lower than those without lymph node metastasis(Z =-2.216,P ＜0.05;t =-2.217,P ＜ 0.05),and so as in the extrathyroid infiltration(Z =-2.267,P ＜ 0.05;t =-3.885,P ＜ 0.01).Conclusion BRAFV600E mutations may accelerate the malignant development of PTC by down regulating the expression of HMGB1.

Objective To explore the mechanism of As2O3 in regulating the apoptosis of anaplastic thyroid cancer (ATC) cell line FRO. Method Cultivated ATC cell line FRO were randomized into 5 groups, as control group, U0126 group (10 μM), low level As2O3 group (1 μM), moderate level As2O3 group (10 μM) and high level As2O3 group (100μM). The expression of apoptotic proteins and ERK signaling transduction proteins were assayed by ELISA. Result The expression of Bax, Caspase-3 and Caspase-6 proteins were significantly increased (P<0.05), while the expression of Bcl2, Ras, Raf MEK and ERK1/2 proteins were decreased (P<0.05) with the treatment of As2O3, and the effects were positively correlated with dose levels (P<0.05). Conclusion As2O3 may significantly promote the apoptosis of anaplastic thyroid cancer cell line FRO, which is possibly correlated with the inhibited overexpression of ERK signaling pathway.

循环肿瘤细胞(circulating tumor cells,CTCs)是指自发地或因诊疗操作由实体瘤或转移灶释放入血的恶性肿瘤细胞.目前,CTCs在恶性肿瘤诊断、分期、评效、治疗、预后中的意义及其富集分离、检测技术已成为研究热点.

目的 探讨桥本甲状腺炎(HT)与ABO血型的关系,丰富HT的病因,为HT的预防及治疗提供更多的依据.方法 参照桥本甲状腺炎的入选标准选择病例组684例,以从中心血站获取的临沂市16 261名汉族人(健康献血者和非自身免疫性甲状腺病患者)的血型资料作为对照组,调查临沂市汉族人HT患者的ABO血型分布,与该地区汉族人ABO血型分布比较,观察A型、B型、AB型、O型在2组人群中的构成比差异,并比较4种血型的人患HT的相对危险度(OR).用玻片凝集法检测ABO血型.用电化学发光法测患者的甲状腺激素(FT3和FT4)、促甲状腺激素(TSH)和相关的甲状腺自身抗体(包括TPOAb、TGAb和TRAb).用SPSS 13.5统计软件包分析数据,构成比的比较用x2检验,OR值比较用Logistic回归分析,P＜0.05为差异有统计学意义.结果 ①临沂市汉族人HT患者的ABO血型分布特点为:O型(35.09％)＞A型(28.07％)＞B型(25.14％) ＞AB型(11.70％),与对照组比较差异存在统计学意义(P＜0.001).②患HT的血型OR为:O(1.3)＞AB(1.2)＞A(1.0)＞B(0.88),O型血者患HT风险显著高于其他血型(P＜0.05).结论 在临沂市汉族人群中,血型为O型者发生HT的风险最高,O型可能是该地区汉族入HT的易感血型.

目的:探讨喉咽鳞状细胞癌组织中细胞角蛋白13(CK13)和细胞角蛋白4(CK4)的表达与喉咽鳞状细胞癌临床病理特征的关系.方法:应用免疫组织化学方法检测56例喉咽鳞状细胞癌患者的癌组织标本及癌旁正常组织中CK13及CK4的蛋白表达,应用VIDAS图像分析仪分析其图像的积分光密度,并进行统计学处理.结果:CK4与CK13在癌旁正常组织中均呈强阳性表达,而在喉咽鳞状细胞癌组织中的表达下降(P＜0.01),且与病理分化程度均呈正相关(P＜0.05).CK4在T1、T2期的表达最强,在T3、T4期的表达依次减弱(F=93.814,P＜0.05);CK4在Ⅰ～Ⅱ期的表达最强,在Ⅲ期、Ⅳ期中的表达逐渐减弱(F=127.894,P＜0.05);CK4在无淋巴结转移的鳞状细胞癌组织中的表达较强,而在有淋巴结转移的鳞状细胞癌组织中表达较弱(T=10.678,P＜0.05).而CK13与肿瘤大小、临床分期及有无淋巴结转移无显著相关.结论:CK4与CK13共同检测对喉咽鳞状细胞癌的早期诊断、手术方法的选择以及判断预后具有指导意义.

Objective: To investigate the expression and clinical significance of the cytokeratin 4 in hypopharyngeal squamous cell carcinomas.Methods: The expression of CK4 was detected by immunohistochemistry approach in normal tissue of latero-carcinoma and tissue of carcinoma in 60 species of hypopharyngeal squamous.Results: The expression of CK4 was strong positive in the normal tissue of latero-carcinoma,while CK4 expression reduced in the hypopharynx squamous carcinoma tissue.The variance was statistical significance(x 2 =73.594,P0.05).Compared with that in the high differentiated squamous cell carcinoma,the expression of CK4 in moderately differentiation and poorly differentiated squamous carcinoma tissue all appear declined obviously.The variance was statistical significance(F=57.082,P0.05).Conclusion: CK4 has low expression in hypopharynx squamous carcinoma.It has certain clinical significance in the diagnosis of hypopharynx squamous carcinoma,and CK4 is expected to become the biological marker of hypopharynx squamous carcinoma malignant degree.

Objective To investigate the expression and clinical significance of cytokeratin 4(CK4) in hypopharyngeal squamous cell carcinoma(SCC).Methods The expression of CK4 in cancer and paraneoplastic tissue of 60 patients with hypopharyngeal SCC was detected by immunohistochemistry method.Results The difference of CK4 expression between cancer tissue and paraneoplastic tissue was significant(χ2=73.594,P0.05),and that in well-,moderately-and poorly-differentiated SCC tissue was also significant(F=57.082;q=7.50-13.02;P0.05).Conclusion Low expression of CK4 in hypopharynx SCC has a certain value in exploring the biological behaviour of hypopharynx SCC,which is expected to become a biological marker to judge the degree of this malignancy.

1临床资料例1女,28岁。因"乏力、怕冷、食欲欠佳11个月,加重伴恶心、呕吐3 d"来诊。患者1年前顺产1胎儿,分娩后无乳汁分泌,产后1个月无明显诱因出现乏力、怕冷、食欲减退,并反复发生"感冒"。3 d前无明显诱因再次出现咽痛、鼻塞、畏寒,测体温35℃,同时伴有明显厌食、恶心及进食后呕吐,化验电解质血钠123 mmol/L、血氯93 mmol/L、血糖3.8 mmol/L;补液治疗后症状及电解质均无明显好转,血皮质醇为0,ACTH 1.16 pg/ml,甲状腺及性腺激素均异常。T:35.0℃,P:72次/min,R:18次/min BP:78/51 mm Hg

<正>病例1,患者女,44岁,2011年1月起出现口干、多饮、消瘦,当地医院诊断"2型糖尿病、结缔组织病",接受四次胰岛素强化治疗,每日总剂量70U左右;同时甲强龙静脉冲击500mg×3d抗免疫、抗炎治疗。血糖不稳定,餐后2h血糖最高25mmol/L,并且经常感晨起心慌、多汗,查空腹血糖最低3.2mmol/L,后自行停用基础胰岛素。入院前15d口渴、多尿症状加重,血糖明显

原发性甲状腺功能减退(甲减)患者血循环中甲状腺激素(thyroid hormone,TH)水平降低,负反馈作用减弱,可致垂体促甲状腺激素(thyroid stimulating hormone,TSH)细胞和催乳素(prolactin,PRL)细胞增生,磁共振示垂体体积增大,临床相对少见,易被误诊为垂体瘤而行手术治疗导致不良后果。现将1例原发性甲减致垂体增生病例报道如下。

目的研究TNF-α诱导的人晶体上皮细胞(HLE-B3)炎症损伤模型中细胞形态及性状的改变,探讨核因子-κB(NF-κB)信号通路在炎症损伤中的作用。方法将HLE-B3细胞分为两组,对照组加入普通培养液,观察组加入含TNF-α的培养液。观察两组细胞形态变化;划痕试验检测细胞迁移率变化;免疫组织化学、Western blot和荧光定量PCR法分别检测NF-κB蛋白及mRNA表达变化;ELISA法检测培养液中NF-κB下游因子IL-1和IL-6水平。结果观察组HLE-B3细胞伸长变细,迁移率明显增快,逐渐向成纤维细胞转化。刺激后NF-κB蛋白表达增多,并从胞质向胞核内转移。与对照组比较,TNF-α刺激后6 h NF-κB蛋白及mRNA表达量均明显增高,12 h时达到高峰;培养液中IL-1和IL-6水平刺激后亦明显增加。结论应用TNF-α进行诱导是建立HLE-B3细胞炎症损伤模型的有效方法;NF-κB信号通路被激活可能是开启炎症损伤的关键。

Objective To explore the clinical significance of typing enterovirus involved in central nervous system(CNS) infection in children. Methods By using reverse transcription polymerase chain reaction(RT-PCR) and consensus primer and typing primer,the enterovirus RNA from cerebrospinal fluid(CSF) of 77(63 acute and 14 convalescence) children with enteroviral CNS infection were detected.The nucleotide sequence of positive specimen was analyzed using VP1 typing primer. Results Of the 77 CSF specimens undergoing RT-PCR with consensus primer amplification,47 out of 63 acute cases(74.6%) were positive,three out of 14 at convalescence stage(21.4%) were positive.These 50 positive specimens were further analyzed with typing primer amplification,31(62.0%) showed positivity.The sequence of 27 randomly selected segments showed 97% nucleotide homology compared with GenBank EV. Conclusion RT-PCR amplification with typing primer and sequence analysis provides a new approach to EV typing.