目的 通过高通量测序法对多重耐药大肠埃希菌HX43进行耐药分子机制的研究。方法 用Illumina Miseq平台对HX43进行高通量测序,用Edena、RAST、ResFinder、MLST和BLAST等生物信息学工具或数据库进行数据分析,获得耐药基因相关序列数据。结果 HX43对多种临床常用抗生素均不敏感,仅对碳氢霉烯类药物敏感。对高通量测序数据的分析研究发现,该菌存在多种耐药基因,包括β-内酰胺类耐药基因3个(bla_CMY-42、bla_CTX-M-14和bla_OXA-30),氨基糖苷类耐药基因5个(aac(3)-IIa、aadA5、 strA、 strB和aac(6′)-Ib-cr),喹诺酮类耐药基因1个(aac(6′)-Ib-cr),磺胺及甲氧苄啶类耐药基因3个(sul1、sul2和dfrA17),四环素耐药基因1个(tet(B)),氯霉素耐药基因2个(catB3和cmlA1),大环内酯类耐药基因2个(erm(B)和mph(A))。对包含bla_CMY-42的contigs进行分析,发现该基因与ISEcp1插入序列、blc和sugE等基因相关联。质粒分型发现HX43携带5种不相容群的质粒。多位点序列分型(MLST)分析发现HX43属于ST3835,为国内外较少见的序列型。结论 高通量测序技术可准确获得临床菌株抗生素耐药的相关基因信息,为临床抗菌治疗提供重要的实验室数据支持。

Objective To investigate the molecular resistance mechanism of Escherichia coli HX43 by high-throughput sequencing. Methods HX43 was sequenced by the Illumina Miseq platform, and sequencing data were analyzed by the Edena, RAST, ResFinder, MLST and BLAST softwares and databases. Results HX43 was resistant to most common clinical antibiotics except carbapenems. Analysis of data revealed resistance genes to β-lactams (bla_CMY-42, bla_CTX-M-14 and bla_OXA-30), aminoglycosides (aac(3)-IIa, aadA5, strA, strB and aac(6′)-Ib-cr), quinolones (aac(6′)-Ib-cr), trimethoprim/sulfonamides(sul1, sul2 and dfrA17), tetracyclines (tet(B)), chloramphenicol (catB3 and cmlA1), macrolides(erm(B) and mph(A)). Sequence analysis of the contig containing bla_CMY-42 identified correlations of the gene with ISEcp1 insertion sequences, blc and sugE genes. Plasmid typing identified 5 plasmid incompatibility groups in HX43. MLST analysis found that HX43 belonged to ST3835, a relatively rare sequence type in the world. Conclusion Information of resistance genes can be obtained by high-throughput sequencing, which provides important experimental data for clinical antimicrobial treatment.

目的 探讨糖尿病胃轻瘫大鼠不同血糖水平对Cajal间质细胞(ICC)的影响及其机制。方法 选择雌性Wista大鼠60只进行随机分组,实验组40只,对照组20只。实验组糖尿病Wista大鼠模型以单次腹腔注射链脲佐菌素法诱导。免疫组织化学荧光染色检测不同血糖浓度大鼠胃ICC数量及网络结构。结果 实验组大鼠血糖浓度高于对照组,ICC数量,低于对照组,且比较差异有统计学意义(P<0.05)。实验组大鼠中血糖浓度越高,ICC数量越低,说明血糖浓度升高可能与平滑肌及神经末梢之间缝隙连接的减少及其ICC网络的超微结构损伤及异常有关。结论 DM小鼠胃组织中血糖水平的升高,可能是DM胃中ICC数量减少的原因;外源性降低血糖能改善DM相关的胃肠道ICC病变。

Objective To observe the effects of glucose fluctuations on Cajal interstitial cells (ICC) of rats with diabetic gastroparesis(DGP) and its mechanistic. Methods 60 Wistar rats were selected and randomly divided into two groups. 20 rats in experimental group and 40 rats in control group. Used immunofluorescence staining to detect the amount of gastric ICC and network structure in DGP rats with different glucose levels. Results The blood glucose concentration in the experimental group was significantly higher than that in the control group, the amount of ICC in the experimental group was significantly lower than that in the control group(P<0.05). The amount of ICC decreased with the increase of glucose levels. In the experimental group, The gap junctions between smooth muscle and nerve endings, ultrastructural damage and abnormalities of the ICC network were probably related to glucose level. Conclusion The increase of glucose level was probably the cause of the decrease of the amount in ICC. Exogenousy decrease glucose levels probably can help to improve the lesion of ICC with DGP.

目的 氧化苦参碱对视网膜母细胞瘤细胞SM-106凋亡的诱导作用及机制。方法 以不同作用时间(24 h、48 h、72 h)和不同作用浓度(12.5 μl/mL、25 μl/mL、50 μl/mL、100 μl/mL)氧化苦参碱处理视网膜母细胞瘤细胞SM-106,分别采用流式细胞仪及western blot检测视网膜母细胞瘤细胞SM-106细胞凋亡及其凋亡因子(Bax、Bcl-2)蛋白表达。结果 氧化苦参碱可促进SM-106细胞体外凋亡,上调Bax蛋白表达及Bax/Bcl-2蛋白表达比值,下调Bcl-2蛋白表达,并呈现剂量及时间依赖性。结论 氧化苦参碱可诱导视网膜母细胞瘤细胞SM-106凋亡,调控凋亡因子Bax、Bcl-2的表达是其可能作用机制。

Objective To evaluate the apoptosis and its mechanism of retinoblastoma cells SM-106 induced by oxymatrine. Methods Retinoblastoma cells SM-106 were treated with different time(24 h、48 h、72 h)and different concentrations(12.5 μl/mL, 25 μl/mL, 50 μl/mL or 100 μl/mL) of oxymatrine. The apoptosis and protein expression of apoptosis factors (Bax and Bcl-2) were respectively determined by flow cytometry and western blot. Results Oxymatrine significantly promoted the SM-106 cells apoptosis in vitro, raised Bax protein expression and Bax/Bcl-2 protein expression ratio, reduced the Bcl-2 protein expression, and showed the dose and time dependent. Conclusion Oxymatrine is able to induce the apoptosis in retinoblastoma cells SM-106. Regulating apoptosis related gene Bax and Bcl-2 expression may be the mechanism of apoptosis.

目的 初步探讨微泡增强的脉冲式超声治疗脾创伤出血的作用机制。方法 14只健康家犬随机分为3组,超声微泡组(MEUS组)6只、单纯超声组(TUS组)4只、单纯微泡组(MB组)4只。开腹切割脾建立脾破裂出血模型,MEUS组用脉冲式超声治疗仪辐照伤口,同时静脉匀速推注微泡;TUS组超声治疗时静脉推注生理盐水;MB组超声治疗仪假照的同时静脉推注微泡。治疗完毕,进行超声造影评价,并送病理组织学检查。结果 MEUS组造影示靶区造影增强缺损或者低灌注,但较粗大血管仍为增强显影。病理组织学见脾窦、微小血管扩张充血、血管周围组织水肿,血小板聚集,微小血管血栓形成。结论 微小血管血栓形成、微血管淤血扩张、周围组织水肿压迫是超声联合微泡治疗脾创伤出血的可能机理。

Objective To investigate the mechanism of haemostatic effect induced by microbubble(MB)enhanced therapeutic ultrasound(TUS)on splenic trauma. Methods 14 healthy dogs were divided into 3 groups.Six animals were treated by microbubble-enhanced therapeutic ultrasound(MEUS),the other eight animals were treated with TUS only group(n=4)and the MB only(n=4)served as the controls.The spleens of all animals were surgically exposed and a 20 mm long,5 mm deep incision was created on the spleens using scalpel.Contrast enhanced ultrasound(CEUS)was performed to assess the blocking effects of splenic circulation.The targeted spleens were harvested for pathological examination. Results A non-enhanced or perfusion defect region was formed within the treated area.The histological results showed splenic sinus hyperemia,microvascular hyperemia,perivascular tissue edema,platelet aggregation and intravascular thrombosis. Conclusion The mechanism of haemostatic effect on splenic trauma by microbubble enhanced ultrasound maybe intravascular thrombosis,microvascular hyperemia and perivascular tissue edema oppression simultaneously.

目的 水解乳清蛋白对炎症性肠病大鼠的抗炎作用及机制。方法 将40只雄性大鼠随机分为实验组和对照组,并建立炎症性肠病动物模型,分别喂食添加了水解乳清蛋白及普通蛋白的饲料,喂养4周后处死大鼠,每周检测体重,血清ALB、TNF-α、IL-2、IL-6等。结果 二组间体重及血清白蛋白无区别(P>0.05),实验组与对照组的TNF-α、IL-2及IL-6无区别(P>0.05),从第二周到第四周,实验组的炎症因子水平低于对照组(P<0.05)。结论 水解乳清蛋白具有抗炎作用,能够减少炎症性肠病大鼠动物模型的炎症因子的释放,并改善其营养状况。

Objective To evaluate the anti-inflammatory effects of whey protein on SD rats model of inflammatory bowel disease. Methods 40 SD rats model of inflammatory bowel disease were established and randomly divided into experimental and control groups equally. Experimental and control groups were fed whey protein and ordinary protein respectively. After 4 weeks, TNF-α, IL-2 and IL-6 were detected. Results There were no significant difference between the two groups of weights and the level of ALB. The level of TNF-α, IL-2 and IL-6 between groups were not significantly different in the first week(P>0.05). However, thelevels of TNF-α, IL-2 and IL-6 in experimental group were significantly lower than those of the control group in the follow weeks. Conclusion The whey protein could reduce the production of inflammatory cytokines.

目的 分析乳腺癌细胞中Snail与MTDH基因的作用,明确Snail是否通过结合于MTDH的启动子区域促进乳腺癌转移。方法 克隆、转染Snail基因至乳腺癌细胞,观察过表达Snail的乳腺癌细胞中MTDHmRNA及蛋白表达的变化;再使用免疫共沉淀法检测Snail与MTDH基因的共作用。结果 转染Snail基因进入乳腺癌MDA-MB-435细胞后,转染组、空白组和对照组中MTDHmRNA的表达水平分别为1.61±0.22、1.02±0.18、0.99±0.20,转染组高于空白组和对照组,差异有统计学意义(P<0.05),而后两组表达无差异(P>0.05);Westren blot检测结果显示,Snail可促进MTDH蛋白的表达;免疫共沉淀显示,Snail与MTDH在细胞内存在相互结合作用。结论 Snail在乳腺癌细胞中可通过结合于MTDH基因的启动子区域,促进MTDHmRNA转录及相关蛋白的表达,从而导致乳腺癌转移。

Objective To investigate the function of Snail to MTDH gene in breast cancer cells. Methods We observed the changement of MTDHmRNA and protein expression in breast cancer cell line MDA-MB-435 after transfected with Snail gene. Then, we used co-immunoprecipitation to determine the domain of Snail and MTDH binding in vitro. Results After transfected with Snail gene into MDA-MB-435 cell, the expression levels of MTDHmRNA in transfection group, blank group and control group were 1.61±0.22,1.02±0.18,0.99±0.20. The level of transfection group was significantly higher than the other groups(P< 0.05). Western blot showed that the expression of MTDH protein can be promoted by Snail. Co-immunoprecipitation showed that Snail and MTDH are binding interactions in breast cancer cell line MDA-MB-43. Conclusion Snail can promote transcription and expression of MTDH in breast cancer cells by binding to the promoter region of the MTDH gene resulting in metastasis of breast cancer.

目的 探讨抗增殖蛋白2（PHB2）脓毒症心肌损伤线粒体功能的调控机制。方法 体外培养大鼠心肌细胞株（H9C2）,分为对照组、脂多糖（LPS）组、LPS+PHB2 siRNA（si-PHB2）组。检测氧化应激指标细胞内丙二醛（MDA）水平、荧光探针检测细胞内活性氧（ROS）水平;线粒体指标：三磷酸腺苷（ATP）水平、线粒体膜电位、线粒体电镜、线粒体半定量评分;免疫印迹法检测PHB2、PTEN诱导激酶1（PTNKI）、帕金蛋白（Parkin）、线粒体转录因子（TFAM）的表达。结果 LPS刺激后MDA水平和ROS水平升高、ATP水平低,LPS+si-PHB2组MDA（6.21±0.39 vs 3.59±0.33, P<0.05）、细胞内的ROS（15 131.37±88.72 vs 8 628.67±71.95, P<0.05）的水平较LPS组升高,ATP（3.46±0.34 vs 4.52±0.25, P<0.05）和线粒体膜电位水平（0.33±0.04 vs 0.55±0.09, P<0.05）进一步降低;电镜观察显示与正常组相比,LPS组、LPS+si-PHB2组出现不同程度线粒体损伤,线粒体损伤半定量评分显示LPS+si-PHB2组的损伤较LPS组更为明显（1.42±0.10 vs 0.81±0.04, P<0.05）; 免疫印迹法结果显示LPS处理后PHB2、PINK1、Parkin 表达上调,TFAM表达下调,LPS+si-PHB2组的线粒体自噬相关蛋白PINK1（1.33±0.06 vs 1.79±0.21, P<0.05）、Parkin（1.43±0.08 vs 1.86±0.09, P<0.05）和线粒体生物发生关键蛋白TFAM（0.29±0.01 vs 0.74±0.06, P<0.05）表达均较LPS组降低。结论 LPS可促进大鼠心肌细胞PHB2表达,si-PHB2干扰后线粒体自噬蛋白和生物发生蛋白表达抑制,心肌细胞氧化应激损害和线粒体功能障碍加重,提示PHB2表达上调可能恢复线粒体稳态改善脓毒症心肌损伤的线粒体功能。

Objective To explore the regulatory mechanism of septic myocardial injury by prohibitin 2（PHB2）. Methods Rat myocardial cell lines（H9C2）were cultured in vitro and divided into control group,LPS group,LPS + PHB2 siRNA（si-PHB2） group. The indicators for detecting oxidative stress include the levels of intracellular malondialdehyde（MDA）and reactive oxygen species（ROS）. The indicators for mitochondrial detection include adenosine triphosphate（ATP）levels,mitochondrial membrane potential,mitochondrial electron microscopy,and semi-quantitative mitochondrial scoring. Western blotting was used to detect the expression of PHB2,PTEN induced putative kinase（PINK1）,Parkin,mitochondrial transcription factor A（TFAM）. Results After LPS stimulation,MDA level and intracellular ROS level increased,ATP level decreased. Compared with LPS group,MDA（6. 21±0. 39 vs 3. 59±0. 33, P<0. 05）level and intracellular ROS level（15 131. 37±88. 72 vs 8 628. 67±71. 95, P<0. 05）in LPS + si-PHB2 group increased significantly,while ATP（3. 46±0. 34 vs 4. 52±0. 25, P<0. 05）and MMP（0. 33±0. 04 vs 0. 55±0. 09, P<0. 05）level further decreased. Compared with the normal group,the structure of mitochondria in LPS group and LPS + si-PHB2 group was damaged in different degree. The semi-quantitative score of mitochondrial damage showed that the damage in LPS + si-PHB2 group was more obvious than that in LPS group（1. 42±0. 10 vs 0. 81±0. 04, P<0. 05）. Western blotting showed that the expression of PHB2,PINK1 and Parkin were up-regulated and the expression of TFAM was down-regulated after LPS treatment,mitohagy-related proteins PINK1（1. 33±0. 06 vs 1. 79±0. 21, P<0. 05）,Parkin（1. 43±0. 08 vs 1. 86±0. 09, P<0. 05）and mitochondrial biogenetic protein TFAM（0. 29±0. 01 vs 0. 74±0. 06, P<0. 05）in LPS+si-PHB2 group were lower than those in LPS group. Conclusions LPS can promote the expression of PHB2 in rat cardiomyocytes. After interfering with PHB2 expression,we found that mitochondrial autophagy and biogenesis are inhibited,and mitochondrial dysfunction,oxidative stress exacerbated,suggesting that the up-regulation of PHB2 expression may restore mitochondrial homeostasis and improve mitochondrial function in septic myocardial injury.

       目的   探讨NXT629改善肝胆结石形成的相关机制。方法   对C57BL/6J小鼠分别采用常规饮食或成石饮食（LD）喂养，并在LD组小鼠注射PPAR-α拮抗剂NXT629。通过苏木精-伊红染色法染色分析肝脂肪病变，油红O染色检测肝脏脂质的积累，分光光度法检测胆汁或血清中总胆固醇、甘油三酯、磷脂、总胆汁酸、胆固醇饱和指数、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇指标；qPCR法检测小鼠肝组织中ABCG5/8、CYP7A1、CYP7B1、PPAR-α和ABCB11 mRNA的表达情况。结果  NXT629通过靶向PPAR-α降低LD组小鼠肝脏中的ABCG5、ABCG8、ABCB11 mRNA水平以及增加CYP7A1、CYP7B1 mRNA水平，进而减少LD诱导的肝胆结石形成并改善脂质代谢紊乱。结论  NXT629可能通过影响脂代谢相关基因表达改善肝胆结石。

       Objective  To explore the mechanism on NXT629 improves hepatolithiasis formation．Methods  C57BL/6J mice were fed either a regular diet or a lithogenic diet（LD），with the LD group receiving injections of PPAR-α inhibitor NXT629．Liver steatosis was analyzed via HE staining，hepatic lipid accumulation was detected by Oil Red O staining，and total cholesterol，triglycerides，phospholipids，total bile acids，cholesterol saturation index，low density lipoprotein cholesterol，and high density lipoprotein cholesterol levels in bile or serum were measured using assay kits．RT-qPCR was employed to determine the mRNA expression of ABCG5/8，CYP7A1，CYP7B1，PPAR-α，and ABCB11 in mouse liver tissues．Results  The results showed that NXT629 target PPAR-α to down-regulate the mRNA levels of ABCG5，ABCG8，and ABCB11 in the livers of LD-fed mice，while increasing the mRNA levels of CYP7A1 and CYP7B1，thereby  reducing LD-induced hepatolithiasis formation and improving lipid metabolism disorders．Conclusions  NXT629 can improve cholesterol gallstones by affecting the expression of genes related to lipid metabolism．

       目的   探讨复方黄芪颗粒（CHG）的抗疲劳作用及其机制。方法  48只雄性BALB/C小鼠随机分为空白对照组、低剂量（9.1 g/kg）、中剂量（18.2 g/kg）、高剂量（27.3 g/kg）CHG 3个试验组，每组12只。试验组给予不同剂量的复方黄芪颗粒溶液灌胃，空白对照组小鼠给予等体积生理盐水。给药30 d后，检测小鼠体内相关指标变化，观察其抗疲劳作用并分析相关机制。结果   与空白对照组相比，试验组小鼠体质量差异无统计学意义（P＞0.05），小鼠力竭游泳时间及转棒耐力时间均明显延长（P＜0.01），血尿素氮（BUN）、乳酸脱氢酶（LDH）、丙二醛（MDA）水平明显降低（P＜0.01），肝糖原和肌糖原水平升高（P＜0.05），超氧化物歧化酶（SOD）活性升高（P＜0.01）。体外抗氧化试验表明CHG以剂量依赖性方式清除2，2-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐（ABTS）和1，1-二苯基-2-三硝基苯肼（DPPH）自由基。当CHG质量浓度为100.000 0 mg/mL时，CHG对DPPH自由基清除能力可达85.030 3%。当CHG质量浓度为25.000 0 mg/mL时，CHG对ABTS自由基清除能力可达96.357 2%。结论  CHG具有抗疲劳的作用，其作用机制可能与抗氧化作用相关。

      Objective  To investigate the anti-fatigue effects of compound Huangqi granules（CHG）and its mechanism．Methods  Forty-eight male BALB/C mice were randomly divided into blank control group，9.1，18.2，27.3 g/kg CHG group（test groups）．The test groups received different concentrations of CHG solution by gavage，and the blank control group mice were given equal volume saline．After 30 days of administration，the mice were tested，meanwhile the anti-fatigue effect and mechanism were investigated．Results  Compared with blank control group，there was no significant difference in body weight（P＞0.05）．The exhaustive swimming time and rod turning endurance time of mice were significantly prolonged（P＜0.01）．The serum levels of blood urea nitrogen，lactate dehydrogenase and malondialdehyde were significantly decreased（P＜0.01），while the liver and muscle glycogen levels（P＜0.05）and superoxide dismutase activity were increased（P＜0.01）．In vitro antioxidant tests showed that CHG can remove （1，1-Diphenyl-2-picrylhydrazyl，ABTS） and （2，2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid，DPPH） free radicals in a dose-dependent manner．When the CHG concentration is 100 mg/mL，the DPPH free  radical scavenging ability of CHG can reach 85.030 3%．When the CHG concentration was 25 mg/mL，the scavenging ability of CHG to ABTS free radicals reached 96.357 2%．Conclusions  CHG has anti-fatigue effect，and its mechanism may be related to anti-oxidation effect．