目的:探索肺腺癌胸水细胞蜡块在肺腺癌病理诊断及EGFR/ALK/ROS1基因突变检测中的应用价值.方法:选取2017年1月至2021年12月在温州医科大学附属第一医院诊断为肺腺癌的41例患者胸水细胞,采用常规细胞涂片与细胞蜡块切片进行病理HE染色鉴定,并采用NGS及ARMS方法进行EGFR/ALK/ROS1基因突变检测.结果:恶性胸腔积液(MPE)细胞涂片癌细胞鉴定敏感性为90.2％,假阴性率为9.8％,MPE细胞蜡块癌细胞鉴定敏感性为100％,假阴性率为0;MPE细胞涂片样本EGFR/ALK/ROS1总体突变率为70.7％(29/41),其中EGFR突变率为61.0％(25/41),ALK融合基因检出率为7.3％(3/41),ROS1融合基因检出率为14.6％(6/41);MPE细胞蜡块样本中EGFR/ALK/ROS1总体突变率为73.2％(30/41),其中EGFR突变率为68.3％(28/41),ALK融合基因检出率为2.4％(1/41),ROS1融合基因检出率为4.9％(2/41);此外,胸水细胞涂片及细胞蜡块的基因突变阳性的肺腺癌患者较未突变肺腺癌患者具有显著增加的总生存率、无进展生存率及疾病控制率.结论:与胸水细胞涂片相比,胸水细胞蜡块肿瘤检出率及EGFR/ALK/ROS1总体突变率相对更高,可用于指导病理诊断及分子检测的靶向治疗.

目的 分析丁苯酞对外伤性蛛网膜下腔出血患者脑循环及血清炎症因子水平的影响.方法 60例外伤性蛛网膜下腔出血患者,按照治疗方式不同分为对照组和观察组,每组30例.对照组应用常规方案治疗,观察组在对照组基础上应用丁苯酞氯化钠注射液治疗.比较两组患者治疗前后血清炎性因子[白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]、脑循环参数[脑血流量(CBF)、脑血容量(CBV)以及平均通过时间(MTT)]及并发症发生情况.结果 治疗后,两组患者IL-1β、IL-6以及TNF-α水平均较本组治疗前降低,且观察组患者IL-1β(2.01±0.36)pg/ml、IL-6(96.34±10.24)pg/ml、TNF-α(5.14±1.17)ng/ml明显低于对照组的(4.15±0.28)pg/ml、(151.24±13.25)pg/ml、(16.35±2.29)ng/ml,差异具有统计学意义(P<0.05).治疗后,两组患者的CBF、CBV较本组治疗前明显升高,MTT较本组治疗前明显缩短,且观察组患者CBF(43.63±1.29)ml/(100 g·min)、CBV(2.29±0.17)ml/100 g均高于对照组的(37.24±1.26)ml/(100 g·min)、(1.86±0.19)ml/100 g,MTT(2.74±0.26)s短于对照组的(3.91±0.27)s,差异具有统计学意义(P<0.05).两组患者并发症发生率比较差异无统计学意义(P>0.05).结论 对外伤性蛛网膜下腔出血患者应用丁苯酞进行治疗有助于降低其血清炎性因子水平,缓解脑循环不良情况,且并发症发生率低,值得在临床推广应用.

目的:探讨高糖环境下miRNA-195抑制剂对血管内皮细胞行为的影响及机制.方法:培养人脐静脉血管内皮细胞(HUVECs)ECV304,分别用高糖(HG)、miRNA-195模拟质粒(mimic)、miRNA-195抑制质粒(inhibitor)、Sirt1激动剂白藜芦醇(RES)处理细胞株,CCK-8试剂盒检测增殖活力,RT-PCR检测miRNA-195及Sirt1 mRNA水平,Western blot检测Sirt1、p-FoxO1、FoxO1、Caveolin-1、血管内皮生长因子(VEGF)蛋白表达水平,划痕实验检测HUVECs的迁移能力,transwell小室实验检测细胞侵袭能力,Matrigel胶检测HUVECs的成管能力.结果:HG促进HUVECs的增殖、迁移、侵袭,增强成管能力(P<0.05);而miRNA-195 inhibitor则明显抑制HUVECs的增殖、迁移,降低成管能力(P<0.05).结论:miRNA-195 inhibitor或许通过miRNA-195/Sirt1/FoxO1/Caveolin-1/VEGF通路影响HG环境下HUVECs的生物学功能.

Objective To investigate the effects of miR-195 on the expression of Sirt1 and its downstream oxidative stress and mitochondrial apoptosis-related proteins in the liver of type 1 diabetic C57BL/6 mice. Methods Mice were divided into control group (Control group), diabetes group (DM group), and miR-195 inhibitor group (DM+antago group). The DM group model was replicated according to the conventional method, and miR-195 inhibitor (2.5 mg/kg) was transfected into DM + antago group mice. The DM group and the control group were injected with scrambled shRNA (2.5 mg/kg) only as a control.Observe and record the increase and decrease of body weight and blood glucose of mice every week. Take fresh liver tissue, one part is processed and made by HE staining for histopathological observation; the part is used to determine the MDA content and T-SOD activity in liver tissue.RT-qPCR and Western blot were used to detect the mRNA and protein content of related indicators in the liver tissue of the three groups of mice. Results 1)Compared with the control group, the leaflets of the liver tissue in the DM group showed structural disorders, hepatocytes were significantly edema, and the cytoplasm was loose; compared with the DM group, the hematocyte edema in the DM+antago group was reduced. 2)Compared with the control group, miR-195 mRNA expression, MDA content and AcFoxo1/Foxo1 protein expression in the DM group were significantly increased (P＜0.05), while Sirt1, Foxo1 mRNA and protein expression, and T-SOD activity were significantly reduced (P＜0.05); Compared with the DM group, miR-195 mRNA expression, MDA content and AcFoxo1/Foxo1 protein expression in the DM + antago group decreased significantly (P＜0.05), while Sirt1, Foxo1 mRNA and protein expression, and T-SOD activity were all Significantly increased (P＜0.05). Conclusions The expression of miR-195 is increased in liver oxidative stress in type 1 diabetic rats. Sirt1 seems to be potential target of miR-195.

目的 观察miR-138对乙醇诱导的大鼠心肌细胞系H9C2的Sirt1表达及其下游氧化应激和线粒体凋亡相关蛋白表达的影响.方法 将心肌细胞分为对照组(control组)、乙醇组(E组)、miR-138抑制剂组(I组);分别用含不同浓度乙醇的细胞培养液培养;MTT法检测H9C2细胞的活力;选择细胞活力接近80％的200 mmol/L乙醇浓度为最佳干预浓度.用Lipo2000,将miR-138抑制剂转染I组H9C2心肌细胞,孵育12 h后,分别用羟胺法测定心肌细胞中总超氧化物歧化酶(T-SOD)活力;硫代巴比妥酸(TBA)法测定心肌细胞丙二醛(MDA)含量;用RT-qPCR检测心肌细胞内miR-138与Sirt1 mRNA的表达;用Western blot测定心肌细胞内Sirt1、Bcl-2、Bax和caspase-3蛋白的表达.结果 1)乙醇可以抑制心肌细胞的活力.2)与对照组相比,乙醇组心肌细胞中miR-138 mRNA的表达及MDA的含量、Bax和caspase-3的蛋白表达均明显升高(P<0.05),T-SOD的活力、Sirt1 mRNA的表达及Sirt1和Bcl-2蛋白的表达均明显降低(P<0.05);与乙醇组相比,miR-138抑制剂组心肌细胞中miR-138 mRNA的表达及MDA的含量及Bax、caspase-3的蛋白表达均明显降低(P<0.05),T-SOD的活力、Sirt1 mRNA的表达、Sirt1和Bcl-2蛋白的表达和均明显升高(P<0.05).结论 miR-138在乙醇诱导的大鼠H9C2心肌细胞氧化应激中表达增加.Sirt1可能是miR-138的作用靶点.

目的 探讨银杏叶提取物(EGB)对1型糖尿病大鼠左心室收缩功能的影响.方法 30只雄性SD大鼠随机分为正常对照组(CON组)、糖尿病模型组(DM组)和银杏叶提取物干预组(EGB组),每组10只.其中后两组用腹腔注射链脲佐菌素(STZ)以诱导1型糖尿病模型.制模成功后,EGB组腹腔注射EGB溶液[8mg/(kg·d)],另两组注射等量0.9%氯化钠注射液.12周后所有大鼠行常规超声心动图及速度向量成像(VVI)检查,测定左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)、乳头肌水平左心室壁各节段收缩期平均峰值速度(Vs)、径向应变(Sr)及径向应变率(SRr),行心导管检查,测定左心室收缩末压力(LVESP)、左心室舒张末压力(LVEDP)及左心室内压最大上升/下降速率(LV±dp/dtmax).处死大鼠后取出心肌组织,分别通过天狼星红染色、TUNEL凋亡染色测定各组大鼠心肌胶原容积分数(CVF)及心肌凋亡指数(AI).结果 EGB干预12周后,DM组大鼠LVEF、LVFS、Vs、Sr、SRr、LVESP、+dp/dtmax及- dp/dtmax明显均低于正常对照组(均P＜0.05),LVEDP、CVF及AI测定值均明显高于正常对照组(均P＜0.05),EGB组LVEF、LVFS、Vs、Sr、SRr、LVESP、+dp/dtmax及- dp/dtmax较DM组均明显增高(均P＜0.05);LVEDP、CVF及AI测定值均明显减低(均P＜0.05).结论 EGB通过减少心肌细胞凋亡,减轻心肌间质纤维化,从而有效改善DM大鼠左心室收缩功能.

目的 观察超声靶向微泡击破(UTMD)技术调控改构型酸性成纤维细胞生长因子(MaFGF)对阿霉素心力衰竭(HF)大鼠心功能的保护作用并探讨其机制.方法 随机将40只实验动物(雄性健康SD大鼠)分为正常对照组、HF模型组、MaFGF组和MaFGF+UTMD组.后3组大鼠通过腹腔注入盐酸阿霉素,MaFGF+UTMD组大鼠经尾静脉注射内含MaFGF的超声微泡混悬液后心脏接受超声靶向微泡击破处理.经过6周干预,超声心动图检查测量左心室收缩末期内径(LVESd)、左心室舒张末期内径(LVEDd)、左心室短轴缩短率(LVFS)以及左心室射血分数(LVEF).其后处死大鼠取心肌组织,检测心肌丙二醛(MDA)、超氧化物歧化酶(SOD)水平,通过Masson胶原染色计算心肌胶原容积分数(CVF)、血管周围胶原面积(PVCA)/血管腔面积(LA)比值,透射电镜下对心肌细胞超微结构进行观察.结果 经过干预6周后,MaFGF+UTMD组LVESd及LVEDd较HF模型组减小(均P<0.05),LVEF、LVFS增高(均P<0.05);HF模型组心肌MDA较正常对照组升高,而SOD降低(均P<0.05),经过MaFGF+UTMD干预,MaFGF+UTMD组SOD上升而MDA降低(均P<0.05);Masson胶原染色显示与正常对照组比较HF模型组CVF与PVCA/LA增高(均P<0.05),而MaFGF+UTMD组这两项指标则下降(均P<0.05);透射电镜显示HF大鼠心脏结构明显异常且能量代谢紊乱,而由UTMD介导的MaFGF干预后,大鼠的心脏微观形态有所改善.结论 对于由阿霉素诱导的心肌损伤,UTMD介导MaFGF对左心室收缩机能具有保护性,其机制可能与MaFGF缓解ROS损害心肌线粒体膜程度,抑制心肌纤维化的形成有关.

目的 探讨姜黄素类似物L6H4对非酒精性脂肪性肝病大鼠肝细胞沉默信息调节因子1 (Sirt1)及凋亡相关蛋白表达的影响.方法 36只SD大鼠随机均分为三组:对照组(C组)、高脂饮食组(F组)和姜黄素组(L组).C组给予基础饲料,F组和L组给予高脂饲料喂养,从第9周开始L组用姜黄素类似物L6H4 0.2 mg/kg灌胃,C组和F组给予相同体积的1％羧甲基纤维素钠灌胃.于第20周末处死,HE染色和Masson染色观察肝组织的病理形态变化,测定丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力以及Sirt1、Bcl-2、Bax和Caspase-3 mRNA及蛋白的表达.结果 C组大鼠肝小叶结构完整,肝细胞呈单层放射状排列;F组大鼠肝小叶结构消失,细胞严重脂肪样变,部分肝细胞坏死伴有炎症细胞浸润;L组大鼠肝小叶结构部分消失,脂肪样变及肝细胞的坏死程度较F组减轻.与C组相比,F组肝组织SOD活力及Sirt1、Bcl-2 mRNA和蛋白表达降低(P＜0.05),MDA含量及Bax、Caspase-3 mRNA和蛋白表达升高(P＜0.05);与F组相比,L组肝组织SOD活力及Sirt1、Bcl-2 mRNA和蛋白表达升高(P＜0.05),MDA含量及Bax、Caspase-3 mRNA和蛋白表达降低(P＜0.05).结论 姜黄素类似物L6H4对非酒精性脂肪性肝病大鼠肝脏具有保护作用;其机制可能是通过调控调节蛋白Bcl-2、Bax和Caspase-3的表达以及上调肝细胞Sirt1的表达来减轻氧化应激而发挥作用.

Nucleotide excision repair (NER), the core mechanism of DNA repair pathway, was commonly used to maintain genomic stability and prevent tumorigenesis. Previous investigations have demonstrated that single nucleotide polymorphisms (SNPs) of NER pathway genes were associated with various types of cancer. However, there was no research elucidating the genetic association of entire NER pathway with ovarian cancer susceptibility. Therefore, we conducted genotyping for 17 SNPs of six NER core genes (XPA, XPC, XPG, ERCC1, ERCC2, and ERCC4) in 89 ovarian cancer cases and 356 cancer-free controls. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to describe the strength of association. The result showed that both ERCC1 rs11615 and XPC rs2228000 were significantly associated with reduced risk of ovarian cancer under dominant genetic model (adjusted OR = 0.35, 95% CI = 0.20–0.61, P=0.0002 and adjusted OR = 0.49, 95% CI = 0.30–0.81, P=0.005 respectively). In addition, XPC rs2228001 and ERCC2 rs238406 had statistically significant association with the increased risk of ovarian cancer under dominant genetic model (adjusted OR = 1.72, 95% CI = 1.02–2.92, P=0.043 and adjusted OR = 2.07, 95% CI = 1.07–4.01, P=0.032 respectively). ERCC1 rs3212986 were related with the increased risk of ovarian cancer under recessive model (adjusted OR = 2.40, 95% CI = 1.30–4.44, P=0.005). In conclusion, our results indicated that ERCC1, XPC and ERCC2 might influence ovarian cancer susceptibility. Further research with large sample size is warranted to validate the reliability and accuracy of our results.

Objective: To explore the roles and mechanism of mitochondrial dynamin-related protein 1 (drp1) and mitochondrial fusion protein 2 (Mfn2) in palmitate (PA) induced rat hepatocyte injury in the model of nonalcoholic-fatty-liver-disease (NAFLD) in vitro, and the effects of curcumin derivative L6H4 on it.Methods: Rat BRL-3A hepatocytes were cultured with difference concentrations of PA and oleate (OA) and PA+ curcumin derivative L6H4 mixed medium for 24 hours respectively. Cell vitality was detected by MTT assay and the opti-mal concentration of PA for establishing non alcoholic fatty liver model in vitro and drug intervention of curcum-in derivative L6H4 were screened. Cells were divided into 4 groups: control group (CON group), oleate group (OA group), palmitate group (PA group) and curcumin derivative L6H4 intervention group (L6H4 group), and were cultured with normal medium, containing oleate medium, containing palmitate medium and containing pal-mitate and curcumin derivative L6H4 mixed medium for 24 hours, respectively. Total SOD activity was detected by hydroxylamine method. The content of MDA was analyzsed by thiobarbituric acid method. The expression of mRNA and protein of Drp1, Mfn2, Bcl-2, Bax, Caspase-3, TNF-α of hepatocyte were detected by real-time PCR and Western blot respectively.Results: Different concentrations of PA had certain inhibitory effect on BRL-3A cells growth (P<0.05), and showed a clear dose dependence manner, 0.1 mmol/L PA was the cut-off point, while 0.05 mmol/L OA had no obvious effect. 0.05 mmol/L PA was selected as the optimal concentration to establish the model of NAFLD in vitro. The cell viability was higher than 75% when the concentration of curcumin deriva-tive L6H4 was lower than 10 μmol/L. The cell viability dropped to below 50% (P<0.05) when the concentration of curcumin derivatives L6H4 was higher than 20 μmol/L, 10 μmol/L L6H4 was the cut-off point. Therefore, 5 μmol/L curcumin derivative L6H4 was chosen for subsequent L6H4 interfere concentration. Compared with the CON group, MDA content of cellular homogenization and the expressions of mRNA and protein of Drp1, Bax, Caspase-3, TNF-α were increased (P<0.05) signiifcantly, T-SOD activity and the expressions of mRNA and pro-tein of Mfn2, Bcl-2 were decreased (P<0.05) signiifcantly in PA group. Compared with the PA group, MDA con-tent of cellular homogenization and the expressions of mRNA and protein of Drp1, Bax, Caspase-3, TNF-α were decreased signiifcantly (P<0.05), T-SOD activity and the expressions of mRNA and protein of Bcl-2 were in-creased signiifcantly (P<0.05) in L6H4 group, but the expression of Mfn2 had no signiifcant difference (P>0.05). Conclusion: Curcumin derivative L6H4 plays an important role in attenuation hepatocyte injury induced by PA, which is associated with its decrease the lipid peroxidation, inhibition mitochondrial ifssion and suppression the downstream of mitochondrial apoptotic pathway and signal transduction of inlfammatory factor. It may be one of the underlying mechanism for curcumin derivative L6H4 to treat NAFLD.

Objective: To investigate the effects of lycopene on bone mineral density (BMD) and RANKL/OPG in chronic hypoxia (CH) modal mice.Methods: Twenty-four C57BL/6 male mice aged 8 weeks were ran-domly divided into three groups: Control group (C group,n=8) and CH group (CH group,n=8) and lycopene group (LCP group,n=8). Mice in CH and LCP were exposed in hypoxic environment, 8 h/day, 6 d/week while the C group lived in normal environment. At the end of 8 weeks, the serum levels of OC and TRACP-5b were mea-sured by ELISA. The content of malondiadehyde (MDA) and the activity of superoxide dismutase (SOD) in shin-bone were analyzed using hydroxylamine assay and TBA colorimetry. The BMD of right femurs were detected by Hologic QDR-4500 bone densitometer. Histomorphology of bone tissue was observed with H.E stainin under light microscopy. The expressions of OPG and RANKL in bone tissue were detected with immunohistochemi-cal technique and RT-qPCR respectively.Results: Firstly, Histologically, the volume of bones trabecular were decreased and the bones trabecular distribution were sparse, thinner and fracture in bone marrow in CH group. In comparison with the C group mice, BMD and serum OC level and the activities of SOD in bone (P<0.01) were decreased signiifcantly in CH group mice, while the serum TRACP-5b level, MDA level in bone, the expres-sion of RANKL protein and RANKL mRNA in bone and RANKL mRNA/OPG mRNA (P<0.01) were increased signiifcantly. Secondly, LCP could partially improve the histomorphometric parameters in mice with CH. The serum TRACP-5b level, MDA level in bone, the expression of RANKL protein and mRNA in bone and RANKL mRNA/OPG mRNA in LCP group mice were lower signiifcantly than the levels in HC group mice, while BMD and serum OC level and the activities of SOD in bone were higher (P<0.01 orP<0.05).Conclusion:Lycopene can reduce oxidative stress, regulate RANKL mRNA/OPG mRNA, decrease bone resorption and improve BMD in CH mice.

目的制备兔骨膜去细胞生物支架,为骨缺损、骨不愈的组织工程研究提供天然的生物支架材料。方法取健康新西兰大白兔,游离双侧胫骨近端内侧骨膜,通过物理冻融（-80℃,24h）、去污剂洗脱（triton-X 100、SDS）和酶消化（DNA酶、RNA酶）获取骨膜去细胞生物支架。通过HE染色、DAPI染色、琼脂糖电泳和基因组DNA定量分析（n=5）测定细胞结构及DNA成分残留;Masson染色和羟脯氨酸测定法（n=6）定性定量检测骨膜细胞外基质的主要成分（胶原）的保留情况;扫描电子显微镜下观察骨膜去细胞生物支架的表面微结构;CCK8法检测支架浸提液毒性;皮下包埋实验（n=4）观察该支架的免疫排斥反应。结果 HE染色和4’,6-二脒基-2-苯基吲哚（DAPI）染色表明去细胞支架无残留细胞;琼脂糖电泳未见明显DNA条带;DNA定量检测显示组织去细胞率达95%以上;Masson染色及羟脯氨酸测定表明去细胞支架胶原成分被保留;扫描电子显微镜下细胞外基质呈现三维网状疏松结构;不同体积分数的浸提液对骨膜细胞的增殖与对照组（普通培养基）比较无明显抑制作用（P＞0.05）;异体皮下包埋实验显示,该去细胞支架免疫排斥反应不明显。结论运用物理冻融、去污剂洗脱和酶消化等方法所获取的骨膜去细胞生物支架细胞去除彻底,细胞外基质的结构及主要成分保留完好,生物相容性良好。

Objective: To investigate the effects of Ginkgo biloba on apoptosis and the expression of Drp-1, Mfn-2, Bax, Bcl-2 and Caspase-3 in mitochondrial apoptotic pathway in neonatal rat myocardial cells induced by high glucose and underlying mechanism.Methods: Left ventricule myocardial cells of neonatal SD rats were isolated and cultured with low-glucose culture media (LG group), low-glucose culture media added EGB (LG +EGB group), high-glucose culture media (HG group) and high-glucose culture media added EGB (HG+EGB group) in vitro respectively. The ratio of apoptosis of myocardial cells was analyzed by lfow cytometry. The ex-pression of Drp-1, Mfn-2, Bax, Bcl-2 and Caspase-3 were detected with western blot and/or RT-PCR respectively. Results: Compared to the LG group, the rate of apoptosis, the expression of Drp-1 and Bax, Caspase-3 and the ratio of Bax/Bcl-2 in ventricule myocardial cells were increased in HG group signiifcantly (P<0.05), while the expression of Mfn-2 and Bcl-2 were decreased signiifcantly (P<0.05). Compared to HG group, the rate of apop-tosis, the expression of Drp-1 and Bax, Caspase-3 and the ratio of Bax/Bcl-2 in HG+EGB group were decreased signiifcantly (P<0.05), while the expression of Bcl-2 was increased signiifcantly (P<0.05). But the expression of Mfn-2 had no signiifcant difference (P>0.05).Conclusion: EGB plays an important role in attenuation cardio-myocyte apoptosis induced by high glucose via its upregulation the expression of Bcl-2 and down regulation of Drp-1 and Bax, Caspase-3 and the ratio of Bax/Bcl-2. It may be one of the underlying mechanisms for EGB to treat diabetic cardiomyopathy.

目的 探讨黄连素(berberine,Ber)对实验性自身免疫性脑脊髓炎(encephalomyelitis,EAE)大鼠脊髓中小胶质细胞及IFN-γ、TNF-α和IL-10水平的影响.方法 Lewis大鼠24只随机分为3组:对照组、模型组和Ber组.采用免疫组化法检测腰髓中小胶质细胞的变化;采用ELISA法检测大鼠外周血和颈髓中促炎因子IFN-γ、TNF-α和抑炎因子IL-10的变化.结果 模型组8只大鼠全部出现典型的EAE行为学改变,Ber组有3只大鼠出现EAE行为学改变.与模型组相比,Ber组大鼠外周血中IFN-γ和TNF-α表达降低(P＜0.05),而IL-10的表达无明显变化;脊髓中活化的小胶质细胞数量减少,IFN-γ和TNF-α表达降低(P＜0.05),IL-10表达增加(P＜0.05).结论 Ber能抑制大鼠EAE的病情,其机制可能与抑制小胶质细胞的活化,下调IFN-γ、TNF-α的表达,并且上调IL-10的表达有关.

Objective To explore the effects of berberine( Ber) on the levels of matrix metallopeptidase-9 ( MMP-9 ) , intercellular cell adhe-sion molecule-1(ICAM-1)in spinal cord of rats with experimental au-toimmune encephalomyelitis. Methods Twenty four Lewis rats were randomly divided into three groups: normal group, model group, Ber group.Neurologic symptoms of Lewis rats were daily examined.Then ti-ssues were stained with Hematoxylin-eosin.Expression levels of MMP-9 and ICAM -1 in spinal cord were quantified by immunohistochemistry. The transcriptional levels of MMP-9,ICAM-1 in spinal cord were exa-mined by real -time polymerase chain reaction ( PCR ) .Results Com-pared with model group, neurological impairment and pathological change were alleviated in Ber group.Protein and mRNA expression of MMP-9 and ICAM-1 were decreased significantly in the spinal cord in Ber group (P<0.01).Conclusion Berberine could have an effect on prevention on EAE in Lewis rat,may be related to its ability to the down regulations the expression levels of MMP-9 and ICAM-1.

Objective To investigate the effects of chronic intermittent hypoxia (CIH) on bone mineral density (BMD) and bone metabolism in mice. Methods Sixteen male C57BL/6 mice aged 12 weeks were randomly assigned into 2 groups:CON group (n=8) and CIH group (n=8). Mice in CIH group were exposed to hypoxic environment 8h ×6d/week, while mice in CON group were exposed to normal environment. At the end of 8 weeks al animals were sacrificed. Serum levels of OC,TRACP- 5b, IL- 6 and TNF- αwere measured by ELISA;histopathology of bone tissue was observed by H.E staining and BMD examination was performed.The expression of OPG and RANKL protein and mRNA in bone tissue was detected by immunohistochemical staining and Real time- qPCR, respectively. Results Histopathology showed that in the metaphysis of distal femurs of CIH mice the bone trabecula became thinner and broken resulting in widening the intertrabecula space. Compared to CON group the BMD, serum OC level and ratio of OPG/RANKL were decreased significantly in the distal femurs of CIH group (P<0.05 or 0.01), while the serum TRACP- 5b,IL- 6 and TNF- αlevel and the expression of RANKL protein and RANKL mRNA in bone were increased significantly (P<0.05 or 0.01). But there were no significant differences in expression of OPG protein and OPG mRNA in bone be-tween two groups(P>0.05). The serum IL- 6 and TNF- αlevels were negatively correlated with BMD(P<0.05 or 0.01), and posi-tively correlated with the expression of RANKL mRNA in bone tissue (P<0.05 or 0.01). Conclusion The results suggest that chronic intermittent hypoxia can increase bone resorption, decrease bone formation and bone mineral density, which are associ-ated with up- regulation of IL- 6 and TNF- αand higher RANKL level.

目的探讨银杏叶提取物（extract of Ginkgo biloba,EGB）对1型糖尿病心肌病大鼠心肌Bcl-2、Bax和caspase-3表达的影响。方法将30只SD雄性大鼠随机分成3组:正常对照组（Con组,n=10）、糖尿病模型组（DM组,n=10）和银杏叶提取物治疗组（EGB组,n=10）。用链脲佐菌素（strepozotocin,STZ）进行腹腔注射,建立1型糖尿病动物模型后用银杏叶提取物注射液腹腔注射建立EGB组,其余两组给予相同体积的生理盐水。于12周末用超声心动图仪测定左心室舒张末期容积（LVIDV）、左心室收缩末期容积（LVISV）、左心室射血分数（LVEF）和每搏射血量（SV）;测定左心室重量指数、血糖和血胰岛素水平;用TUNEL法测定心肌细胞的凋亡指数（AI）;用RT-PCR和Western blot法分别检测其Bcl-2、Bax和caspase-3的表达。结果与Con组相比,DM组大鼠LVIDV（P＜0.01）、SV（P＜0.01）、胰岛素含量（P＜0.01）和心肌组织Bcl-2（P＜0.05）的表达都显著降低;而左心室重量指数和血糖浓度（P＜0.01）、心肌组织中Bax（P＜0.05）、caspase-3（P＜0.05）的表达以及Bax/Bcl-2比例均明显升高。当EGB干预治疗后,与DM组相比,大鼠LVIDV（P＜0.05）、SV（P＜0.05）、胰岛素含量（P＜0.05）和心肌组织Bcl-2（P＜0.05）的表达均显著升高;而左心室重量指数（P＜0.05）、血糖浓度（P＜0.01）、心肌组织中Bax（P＜0.05）、caspase-3（P＜0.05）的表达以及Bax/Bcl-2比例（P＜0.05）均明显降低。但是TUNEL染色显示各组大鼠的心肌细胞凋亡率（cardiomyocyte apoptosis index,CAI）没有明显的差异（P＞0.05）。结论银杏叶提取物可以通过增加Bcl-2的表达,降低Bax、caspase-3表达及Bax/Bcl-2比例,减少心肌细胞的凋亡,从而提高了糖尿病心肌病的心肌功能。这也提示EGB通过调节心肌凋亡相关基因的表达,减少细胞的凋亡是其治疗糖尿病心肌病的重要机制之一。

Objective:To investigate the effects of inlfammatory mediator and oxidative stress on bone me-tabolism in high fat diet-induced obesity mice and the underlying mechanism. MethodSixteen C57BL/6 mice aged 5 weeks were randomly divided into two groups. High fat diet group (HFD) was fed with high fat diet while normal fat diet group (NFD) was fed with normal fat diet. At the end of 16 weeks, the visceral fat was weighed and the visceral fat/body mass ratio was assessed. The levels of TRACP-5b, OC, IL-6 and TNF-αin serum were measured by ELISA. The content of malondiadehyde (MDA) and the activity of superoxide dismutase (SOD) in shin-bone were analyzed by using hydroxylamine assay and TBA colorimetry. The bone mineral density (BMD) of right femurs were detected by Hologic QDR-4500 Bone Densitometer. The expressions of OPG and RANKL in bone tissue were detected by immunohistochemical technique and RT-qPCR respectively. Histologi-cal changes in the left femurs were studied with HE stained under light microscopy. ResultCompared to the NFD group mice, the body weight,the ratio of visceral fat/body mass and the RANKL/OPG in bone, the serum levels of TRACP5b, IL-6, TNF-α, the expressions of RANKL and the content of MDA in bone were increased in HFD group mice signiifcantly (P<0.05). But the serum level of OC, the expressions of OPG, the activity of SOD in bone and the BMD were decreased in HFD group mice signiifcantly (P<0.05). Histologically, the volume of bones trabecular were decreased and the bones trabecular distribution were sparse, thinner and fracture and more adipocytes inifltrated in bone marrow in HFD group. The results of correlation analysis shows that BMD was positively correlated with SOD of bone (r=0.627, P<0.01) and negatively correlated with the ratio of visceral fat/body mass (r=-0.858, P<0.01), MDA level of bone (r=-0.538, P<0.01);SOD was positively correlated with OPG mRNA (r=0.637, P<0.01) and negatively correlated with TNF-α(r=-0.673, P<0.01), IL-6 (r=-0.874, P<0.01), RANKL mRNA (r=-0.760, P<0.01) and the ratio of the RANKL/OPG in bone(r=-0.768, P<0.01), but MDA shows no correlation with TNF-α, IL-6, OPG mRNA, RANKL mRNA and the ratio of the RANKL/OPG in bone (P>0.05). Conclusion:High fat diet induced obesity can lead to the increase of bone resorption and the patho-logical histology changes of osteoporosis and the BMD decline. The underlying mechanism may be associated with its upreguating of inlfammatory mediator and enhancing oxidative stress in bone.

目的:研究银杏叶提取物（EGB）对I型糖尿病心肌病大鼠心肌TGF-β1和collagen表达及间质纤维化的影响。方法:（1）在体实验,取30只雄性SD大鼠随机分为正常对照组（CON组）、糖尿病组（DM组）和EGB干预组（EGB组）;用链脲佐菌素诱导I型糖尿病大鼠模型,于第12周末测定大鼠体重、左心室重量、血糖、糖化血红蛋白和血胰岛素;用超声心动图仪检测左室舒张末期容积（LVEDV）、左室收缩末期容积（LVESV）、左室射血分数（LVEF）和每搏射血量（SV）;用天狼星红染色、免疫组织化学染色及RT-PCR方法分别检测左心室心肌胶原含量、转化生长因子β1（TGF-β1）蛋白表达及TGF-β1、procollagen I和collagen Ⅲ mRNA表达。（2）离体实验,分离培养SD大鼠乳鼠心肌细胞并分别用低糖（LG组）、高糖培养基（HG组）和EGB干预（HG+EGB组）;RT-PCR方法检测各组心肌细胞TGF-β1、procollagen I和collagen Ⅲ的mRNA表达。结果:（1）与CON组相比,DM组大鼠血糖（P＜0.01）、糖化血红蛋白（P＜0.05）、左心室重量指数（P＜0.01）、心肌间质胶原纤维含量（P＜0.05）以及心肌组织内TGF-β1（P＜0.05）、procollagen Ⅰ（P＜0.05）和collagen Ⅲ（P＜0.05）表达均明显升高,而血胰岛素水平（P＜0.01）、LVEDV（P＜0.01）和SV（P＜0.01）明显降低;EGB干预治疗后,与DM组相比,EGB组糖尿病大鼠血糖（P＜0.01）、糖化血红蛋白（P＜0.05）、左心室重量指数（P＜0.05）、心肌间质胶原纤维含量（P＜0.05）以及心肌组织内TGF-β1（P＜0.05）、procollagen Ⅰ（P＜0.05）和collagen Ⅲ（P＜0.05）表达均明显降低,血胰岛素水平（P＜0.05）、LVEDV（P＜0.05）和SV（P＜0.05）明显升高。（2）与LG组比较,HG组心肌细胞TGF-β1（P＜0.01）、procollagen Ⅰ（P＜0.01）和collagen Ⅲ（P＜0.01）mRNA表达明显升高;而EGB干预后,与HG组比较,EGB组心肌细胞TGF-β1（P＜0.01）、procollagen Ⅰ（P＜0.01）和collagen Ⅲ（P＜0.01）mRNA表达明显降低。结论:EGB可通过调低糖尿病大鼠心肌组织TGF-β1的表达,减少心肌间质中collagen Ⅰ和Ⅲ合成及沉积,从而改善糖尿病心肌病大鼠早期的心肌纤维化及左心室舒张功能。这提示通过有效降低心肌TGF-β1的表达减少心肌间质纤维化可能是EGB防治糖尿病心肌病的重要机制之一。

目的 探讨3.0T MR 体外相控阵线圈MRS在慢性前列腺炎的表现及其病理基础.方法 分析经手术或穿刺活检证实的7例表现为MR T2WI低信号的慢性前列腺炎的MRI、MRS所见.所有病例均应用3.0T MR 体外相控阵线圈行MR常规检查、扩散加权成像及MRS检查.MRS测量前列腺T2WI低信号区及对侧正常区域的枸橼酸盐(Cit)、胆碱(Cho)、肌酸(Cr)的相对值及( 胆碱+肌酸) /枸橼酸盐[(Cho+Cr)/Cit]的比值,并计算T2WI低信号病变区与对侧正常区域的胆碱比值及枸橼酸盐比值.分析病变区域的病理镜下所见.结果 慢性前列腺炎病变区域与对侧正常区域的胆碱含量比值为1.05±0.23,病变区域与对侧正常区域的枸橼酸盐含量比值为0.31±0.14,病变区域的(Cho+Cr)/Cit比值(1.77±0.41)明显高于对侧正常区域的(Cho+Cr)/Cit比值(0.58±0.15),两者有明显统计学差异(P＜0.01).病理上,慢性前列腺炎病变区域见淋巴细胞浸润、腺泡及导管不同程度萎缩、闭塞.结论 在常规MR T2WI上表现为低信号的慢性前列腺炎,在MRS上(Cho+Cr)/Cit比值升高,与前列腺癌表现极为相似,但慢性前列腺炎的(Cho+Cr)/Cit比值升高主要是因为病变区域的枸橼酸盐含量降低,而其胆碱含量变化不明显.