GLP-1类似物利拉鲁肽对糖尿病神经病变大鼠坐骨神经PARP-1/NF-κB表达的影响

目的：　通过构建大鼠糖尿病周围神经病变（diabetic peripheral neuropathy,DPN）模型探究胰高血糖素样肽1（glucagon-like peptide 1,GLP-1）类似物利拉鲁肽对DPN大鼠坐骨神经聚腺苷二磷酸核糖聚合酶/激活核因子-κB（poly ADP-ribose polymerase/nuclear factor kappa-B,PARP/NF-κB）表达的影响。方法：　SD大鼠84只,随机选12只作为正常组;其余大鼠均采用链脲佐菌素诱导建立DPN模型。建模成功后,抽取72只大鼠随机分为模型组,胰岛素组,甲钴胺组,利拉鲁肽低、中、高剂量组。正常组和模型组皮下注射等体积生理盐水,胰岛素组给予胰岛素治疗,甲钴胺组给予甲钴胺治疗,给药组给予不同剂量利拉鲁肽治疗,共治疗8周。记录各组大鼠体质量和血糖,测定神经反应速度,ELISA检测炎症因子,流式细胞术检测细胞凋亡,Western blot检测PARP-1、NF-κB蛋白表达,EMSA检测NF-κB转录活性。选择RSC96细胞建立高糖环境下细胞损伤模型,应用基因芯片预测PARP-靶基因,并通过转染PARP-1进行检验。结果：　与模型组比较,利拉鲁肽治疗后大鼠体质量显著增加,血糖显著降低,神经反应速度提高（P<0.05）;大鼠实验和细胞实验显示利拉鲁肽给药后细胞炎症因子含量及PARP和NF-κB蛋白表达较模型组显著降低,高剂量组降低幅度更明显（P<0.05）;大鼠神经细胞凋亡率较模型组显著降低（P<0.05）;利拉鲁肽抑制了NF-κB的转录活性;PARP-1转染组炎症因子和PARP-1、NF-κB蛋白水平显著高于中剂量组（P<0.05）,PARP-1/NF-κB是GLP-1对糖尿病神经病变大鼠抗炎症作用的重要靶点。结论：　利拉鲁肽可以降低DPN大鼠坐骨神经PARP-1和NF-κB的表达水平,从而降低大鼠坐骨神经损伤,起到防治糖尿病周围神经病变损伤的作用。     

BDNF联合chABC鞘内注射对大鼠脊髓损伤修复的影响

目的 探究脑源性神经营养因子（BDNF）联合软骨素酶（chABC）鞘内注射对大鼠脊髓损伤后运动感觉功能的影响以及脊髓组织中神经干细胞、神经元、星形胶质细胞、少突胶质细胞的影响。方法 将SD大鼠分为假手术对照组（A组）,损伤对照组（B组）,chABC治疗组（C组）,BDNF治疗组（D组）,联合治疗组（E组）,采用后肢运动功能评分法（CBS）对大鼠术后1周、2周、4周和2个月后肢运动功能进行评定,苏木精-伊红（HE）染色法观察不同时间点大鼠脊髓组织变化,免疫组化法观察各组大鼠脊髓组织神经干细胞的影响,荧光免疫组化法观察各组大鼠神经元、星形胶质细胞、少突胶质细胞的变化。结果 术前各组大鼠神经功能均正常,术后1~2周,B、C、D、E各组CBS评分降低,但各组间对比差异无统计学意义（P>0.05）;术后4周~2个月,C、D、E组CBS评分逐渐降低,低于B组,差异有统计学意义（P<0.05）。与A组相比,B组不同时间点BrdU阳性细胞数目降低,差异有统计学意义（P<0.05）。脊髓损伤后1周,C、D、E组大鼠脊髓组织BrdU阳性细胞数目与B组比较,差异无统计学意义（P>0.05）;脊髓损伤后第2~4周,C、D、E组BrdU阳性细胞数高于B组,差异有统计学意义（P<0.05）。与空白对照A组比较,B组脊髓损伤后不同时间点Nestin阳性细胞数目均减少,差异有统计学意义（P<0.05）。脊髓损伤后2周、4周、2个月,C、D、E组脊髓组织Nestin阳性细胞数高于B组,差异有统计学意义（P<0.05）。脊髓损伤后1周,A组星形胶质细胞比例逐渐降低,神经元细胞比例逐渐升高。B组细胞3种细胞比例均较低,随着时间延长,神经元细胞比例缓慢升高,星形胶质细胞比例缓慢下降。C、D、E组脊髓受损后脊髓受损后4周~2个月,C、D、E组神经元比例逐渐升高,星形胶质细胞逐渐下降,与B组比较,差异有统计学意义（P<0.05）。结论 BDNF、chABC鞘内注射后对大鼠脊髓损伤后运动、感觉功能恢复具有一定的促进作用,可能与促进神经干细胞增殖以及神经元细胞的分化有关,但两者联合后效果并不显著。     

Z-藁本内酯对人脐静脉内皮细胞保护作用及其机制研究

目的 通过观察Z-藁本内酯对肿瘤坏死因子-α（TNF-α）诱导的人脐静脉内皮细胞（HUVEC）细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）及核因子-κB（NF-κB）表达的影响,探讨其对炎性因子损伤内皮细胞的保护作用及其机制。方法 向体外培养的HUVEC中加入TNF-α 10 ng/mL造成细胞损伤,同时加入不同浓度（5、10、20 μmol/L）的Z-藁本内酯,共同孵育24 h,MTT法检测各组细胞活力,ELISA法测定细胞培养液中ICAM-1和VCAM-1的量,Western blotting法检测各组细胞NF-κB蛋白表达。结果 与对照组比较,TNF-α可造成HUVEC明显损伤（P＜0.05、0.01）,显著增加HUVEC中ICAM-1和VCAM-1的表达（P＜0.01）。与模型组比较,Z-藁本内酯可以剂量相关地降低ICAM-1（P＜0.01）和VCAM-1（P＜0.05）的表达,显著减少细胞NF-κB蛋白的表达（P＜0.05）。结论 Z-藁本内酯可减轻TNF-α对HUVEC的损伤,其作用机制可能与通过抑制NF-κB的激活,进而减少ICAM-1和VCAM-1的表达有关。     

苓桂术甘汤对慢性心衰模型大鼠心肌组织TNF-α及血清NF-κB和IL-1β的影响

目的 观察苓桂术甘汤对慢性心力衰竭模型大鼠心肌组织肿瘤坏死因子-α（TNF-α）蛋白及mRNA表达、血清核因子-κB（NF-κB）、白细胞介素-1β（IL-1β）水平的影响,探讨苓桂术甘汤防治慢性心衰的作用机制。方法 采用冠状动脉结扎法制备慢性心衰大鼠模型,造模4周后将模型大鼠随机分为模型组,卡托普利（4.375 mg/kg）阳性对照组,苓桂术甘汤低、中、高剂量（生药2.1、4.2、8.4 g/kg）组,另设假手术组,每天给药1次,连续给药4周。Western blotting、RT-PCR法分别检测各组大鼠心肌组织TNF-α蛋白及mRNA的表达,ELISA法检测血清中NF-κB、IL-1β水平。结果 与假手术组相比,模型组大鼠心肌组织TNF-α蛋白及mRNA表达增强,血清NF-κB、IL-1β水平显著升高（P＜0.01）;与模型组相比,苓桂术甘汤及卡托普利均能显著抑制模型大鼠心肌组织TNF-α蛋白及mRNA表达、降低模型大鼠血清NF-κB、IL-1β水平（P＜0.05、0.01）。结论 苓桂术甘汤干预慢性心衰的机制与其调节细胞因子网络有关。     

姜黄素对肺炎链球菌感染的抗炎和抑菌作用

目的 研究姜黄素（Cur）及其衍生物6B、Y20对肺炎链球菌（streptococcus pneumonia,SP）最小抑菌浓度及对其致病因子NanA、NF-κB表达调节,探讨Cur、6B、Y20对SP抑菌作用及减少SP感染炎性反应的作用机制。方法 利用微量浓度稀释法联合血琼脂平板涂布法检测Cur、6B、Y20对SP的最小抑菌浓度（minimum inhibitory concentration,MIC）值;培养人支气管上皮（BEAS-2B）,建立体外SP感染模型,分为空白对照组、SP感染组、Cur+SP组、6B+SP组、Y20+SP组。CCK8法测试细胞存活率,ELISA法检测NanA、TNF-α等的蛋白含量,用Western blot法检测NF-κB的表达。结果 Cur、6B、Y20在高浓度有抑菌作用;随SP感染时间的延长,药物处理组细胞存活率较SP感染组相比增高（P<0.01）;与空白对照组比较,SP感染组中NanA、NF-κB、TNF-α表达增加;与SP感染组比较,Cur、6B、Y20药物预处理组NF-κB、NanA表达降低（P<0.05）,TNF-α等表达水平较模型降低（P<0.01）。结论 Cur、6B、Y20高浓度有抑菌作用,并可通过下调SP表面毒力因子NanA的表达,减轻SP的毒力作用,并通过抑制NF-κB转录激活,减少下游炎性因子表达,减轻SP引起炎症损伤。     

通肺化痰汤加减对痰热郁肺型COPD急性加重期患者血清LCN1和LCN2水平的影响

[目的] 观察通肺化痰汤加减对痰热郁肺型慢性阻塞性肺疾病急性加重期（AECOPD）患者的临床疗效及对血清LCN1和LCN2水平的影响。[方法] 选取痰热郁肺型AECOPD患者80例,应用随机数字表法分为对照组和治疗组（每组40例）,两组患者均应用吸氧、抗感染等基础治疗,对照组使用茶碱缓释片、沙美特罗替卡松粉吸入剂,治疗组在对照组基础上加用通肺化痰汤加减,治疗结束后比较两组患者临床疗效、肺功能、肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-8及脂质运载蛋白（LCN）1、LCN2水平的变化。[结果] 治疗后,治疗组总有效率（90%）高于对照组总有效率（72.5%,P<0.05）,两组肺功能水平较治疗前升高,治疗组高于对照组（P<0.05）;治疗后TNF-α、IL-8水平较治疗前降低,治疗组低于对照组（P<0.05）;治疗后LCN1、LCN2水平较治疗前降低,治疗组低于对照组（P<0.05）,治疗过程中未发生不良反应。[结论] 通肺化痰汤对治疗痰热郁肺型AECOPD患者疗效显著,安全性高,可明显改善患者肺功能,降低血清TNF-α、IL-8、LCN1、LCN2水平,提高临床疗效。     

黄芪甲苷抑制IKK/NF-κB炎症通路减轻高糖诱导的H9c2心肌细胞损伤

[目的]观察黄芪甲苷减轻高糖诱导H9c2细胞损伤的作用机制。[方法]通过高糖孵育诱导H9c2心肌细胞损伤模型,MTT法测定细胞存活率,酶联免疫吸附（ELISA）法检测细胞培养液上清中肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的含量,蛋白印迹（Western Blot）法检测细胞内IκB激酶β（IKK-β）、核转录因子κB（NF-κB）、p-NF-κB及TNF-α蛋白表达水平。[结果]给予高糖处理H9c2心肌细胞12 h能明显下调细胞存活率（P<0.05）;黄芪甲苷（20、40、80 μmol/L）预处理10 min可呈剂量依赖性增强细胞活力,降低细胞培养液上清中TNF-α和IL-6的含量（P<0.05）,下调IKK-β蛋白表达（P<0.05）,抑制NF-κB磷酸活化水平（P<0.05）,并降低TNF-α蛋白表达（P<0.05）。[结论]黄芪甲苷可通过抑制IKK/NF-κB炎症通路过度激活减轻高糖诱导的H9c2心肌细胞损伤。     

抵挡汤早期干预对2型糖尿病大鼠大血管病变的影响及其机制

目的 观察抵挡汤早期干预对糖尿病大鼠大血管病变的影响及其机制。方法 采用高脂饲料喂养及链尿佐菌素（STZ）诱导方法制备2型糖尿病大鼠模型,大鼠分成对照组,模型组,抵挡汤早期（给予STZ前4周给药）、中期（与STZ同时给药）及晚期（给予STZ后4周给药）干预组（每天均ig给予抵挡汤3.24 g/kg 1次）,阳性药吡咯列酮（2.7 mg/kg）和辛伐他汀（1.8 mg/kg）组,各组给药至给予STZ后24周。免疫组化染色法观察大鼠主动脉细胞间黏附分子-1（ICAM-1）和血管细胞黏附分子-1（VCAM-1）的表达,Western blotting法检测主动脉核因子-κB（NF-κB）及基质金属蛋白酶-9（MMP-9）蛋白表达。结果 与模型组比较,抵挡汤早期和中期干预组、辛伐他汀组大鼠主动脉ICAM-1表达减少（P＜0.05）;抵挡汤早期干预组与辛伐他汀组主动脉VCAM-1表达减少（P＜0.05）;抵挡汤早期和中期干预组主动脉NF-κB及MMP-9蛋白表达明显降低（P＜0.05）,其中以抵挡汤早期干预组效果最显著。结论 抵挡汤早期干预有效降低NF-κB调控的靶基因ICAM-1、VCAM-1及MMP-9蛋白表达,可能是其延缓2型糖尿病大鼠大血管病变发展的重要机制之一。     

蝎毒提取物对Lewis细胞及DU-145细胞转移的抑制作用

目的 观察蝎毒提取物对Lewis肺癌细胞肺转移和前列腺癌细胞株DU-145骨转移的抑制作用。方法 C57BL/6小鼠尾iv Lewis肺癌细胞（实验分模型组和给药组）,裸鼠尾iv CM-DiI标记DU-145细胞（实验分模型组、对照组和给药组）,给药组均每天ig给予蝎毒提取物300 mg/kg 1次,C57BL/6小鼠给药12 d,裸鼠给药24 d,模型组和对照组给予等量生理盐水,肉眼及镜检观察肺转移灶Lewis肺癌细胞肺转移情况,活体成像系统观察DU-145细胞骨转移的情况。在体外实验,免疫组化法检测Lewis细胞基质金属蛋白酶-9（MMP-9）的表达,免疫组化法和Western blotting检测DU-145细胞MMP-9、NF-κB、基质金属蛋白酶抑制因子1（TIMP-1）、IκB-α蛋白表达。结果 接种Lewis肺癌细胞后,蝎毒提取物组平均癌细胞肺转移灶数为3.2±1.2,而模型组为13.5±5.3,差异显著（P＜0.01）。活体成像检测显示,接种DU-145细胞30 d后,对照组未见荧光团,模型组8只裸鼠中有6只肩胛骨、骨盆、肋骨及胫骨显示有大量荧光团存在,蝎毒提取物组仅有3只裸鼠出现明显荧光团。免疫组化法和Western blotting检测显示,与对照组相比,蝎毒提取物给药后DU-145细胞MMP-9、NF-κB蛋白表达下调（P＜0.01）,TIMP-1表达上调（P＜0.01）,免疫组化检测亦显示IκB-α蛋白表达上调（P＜0.05）。结论 蝎毒提取物抑制Lewis肺癌细胞肺转移和DU-145细胞骨转移,其作用是通过NF-κB和TIMP-1途径抑制MMP-9表达实现的。     

MicroRNA-146a对急性胰腺炎胰腺腺泡细胞增殖及凋亡的影响及其机制研究

目的 探讨microRNA-146a（miR-146a）对急性胰腺炎胰腺腺泡细胞增殖及凋亡的影响，并分析相关机制。方法 体外诱导并培养急性胰腺炎MPC-83细胞，将急性胰腺炎MPC-83细胞分为阴性对照组（mimics-NC组）、过表达miR-146a组（miR-146a-mimics组），另以未经诱导处理的MPC-83细胞为空白对照组（NG组）。采用实时荧光定量聚合酶链反应检测各组MPC-83细胞miR-146a，MTT法检测细胞增殖情况，流式细胞仪检测细胞凋亡情况，酶联免疫吸附试验检测肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6），Western blotting检测增殖细胞核抗原（PCNA）、Bcl-2相关X蛋白（Bax）、B淋巴细胞瘤2（Bcl-2）、核转录因子-κB p65（NF-кB p65）蛋白的表达。结果 急性胰腺炎细胞模型建立成功，成功转染后，与NG组、mimics-NC组比较，miR-146a-mimics组细胞凋亡率，TNF-α、IL-6、Bax和NF-κB p65蛋白相对表达量降低（P <0.05），细胞存活率、PCNA和Bcl-2蛋白相对表达量升高（P <0.05）。结论 过表达miRNA-146a可抑制急性胰腺炎MPC-83细胞凋亡并促进细胞增殖，其作用可能通过抑制NF-кB通路活化来实现。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.