1,25-二羟维生素D3对胃癌细胞增殖和TNF-α表达的影响

目的研究1,25-二羟维生素D3[1,25(OH)2D3]对胃癌SGC-7901细胞增殖和细胞周期的影响,以及对细胞因子TNF-αmRNA及蛋白表达的影响。方法用不同浓度1,25(OH)2D3处理胃癌SGC-7901细胞,采用MTT法检测细胞增殖情况;流式细胞仪检测细胞生长周期;RT-PCR和Western blotting分别检测TNF-α在mRNA和蛋白水平的表达情况。结果 1,25(OH)2D3对胃癌SGC-7901细胞增殖有抑制作用(P<0.05),并呈时间剂量依赖性。经1,25(OH)2D3处理72 h后细胞周期出现向G0/G1期移行的动力学改变(F=9.81,P<0.05)。TNF-αmRNA及蛋白的表达水平均呈剂量依赖性减少(P<0.05)。结论 1,25(OH)2D3可抑制胃癌细胞增殖,且与其抑制TNF-α的表达可能有相关性。     

人牙周膜细胞骨保护因子和破骨细胞分化因子蛋白的表达及1α,25(OH)2维生素D3的调节

目的:研究人牙周膜细胞骨保护因子(osteoprotegerin,OPG)和破骨细胞分化因子(receptor activator nuclear factor kappa B ligand,RANKL)蛋白的表达,以及骨吸收促进因子1α,25(OH)2维生素D3对OPG蛋白表达的影响.方法:用系列酶消化法体外培养人牙周膜细胞,用免疫细胞化学检测细胞上表达的RANKL蛋白,以酶联免疫吸附实验检测无外源性刺激以及1α,25(OH)2维生素D3诱导2、4、6 d时细胞分泌到培养基中的OPG蛋白.结果:人牙周膜细胞胞膜和胞浆上表达RANKL蛋白,分泌OPG蛋白到培养基中.1α,25(OH)2维生素D3降低OPG蛋白的分泌.结论:人牙周膜细胞表达OPG和RANKL,1α,25(OH)2维生素D3影响OPG的表达,推测其通过OPG/RANKL通路参与骨改建.这一结论为进一步研究牙周膜细胞参与骨改建的机制打下基础.     

1,25-二羟维生素D3对胃癌细胞增殖和TNF-α表达的影响

摘要：目的研究1,25-二羟维生素D3[1,25(OH)2D3]对胃癌SGC-7901 细胞增殖和细胞周期的影响,以及对细胞因子TNF-α
mRNA及蛋白表达的影响。方法用不同浓度1,25(OH)2D3处理胃癌SGC-7901细胞,采用MTT法检测细胞增殖情况;流式细胞
仪检测细胞生长周期;RT-PCR和Western blotting分别检测TNF-α在mRNA和蛋白水平的表达情况。结果1,25(OH)2D3对胃癌
SGC-7901细胞增殖有抑制作用（P<0.05）,并呈时间剂量依赖性。经1,25(OH)2D3处理72 h后细胞周期出现向G0/G1期移行的动
力学改变（F=9.81,P<0.05）。TNF-α mRNA及蛋白的表达水平均呈剂量依赖性减少（P<0.05）。结论1,25(OH)2D3可抑制胃癌
细胞增殖,且与其抑制TNF-α的表达可能有相关性。
     

3种非甾体抗炎药对大鼠骨关节炎关节软骨细胞IL-1β、TNF-α、IL-1α表达的影响

目的:探讨长期应用塞来昔布、布洛芬、吲哚美辛对大鼠骨关节炎(osteoarthritis,OA)关节软骨细胞白细胞介素-1β(in-terleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、细胞白细胞介素-1α(interleukin-1α,IL-1α)表达的影响。方法:采用左侧跟腱切除术建立Wistar大鼠同侧膝关节OA模型,分4组,每日给赛来昔布24 mg/kg(celecoxib,CE)组、布洛芬72 mg/kg(ibuprofen,IBP)组、吲哚美辛9 mg/kg(indomethacin,IN)组及生理盐水(normal saline,NS)组。用免疫组织化学法观察3、6、9个月后关节软骨细胞IL-1β、TNF-α、IL-1α表达的变化。结果:①连续用药3月,CE对软骨细胞IL-1β、TNF-α表达无明显影响,抑制IL-1α的表达;IBP促进软骨细胞IL-1β、TNF-α的表达,对IL-1α的表达无明显影响;IN轻度促进软骨细胞IL-1β表达,增强TNF-α的表达,对IL-1α的表达无明显影响。②连续用药6月,CE对软骨细胞IL-1β表达无明显影响,轻度抑制TNF-α、IL-1α表达;IBP轻度促进软骨细胞IL-1β的表达,促进TNF-α表达,轻度抑制IL-1α的表达;IN轻度抑制软骨细胞IL-1β,抑制TNF-α表达,轻度抑制IL-1α的表达。③连续用药9月,CE明显抑制软骨细胞IL-1β的表达,抑制TNF-α、IL-1α的表达;IBP抑制软骨细胞IL-1β、TNF-α的表达,轻度抑制IL-1α的表达。结论:CE抑制软骨细胞IL-1β、TNF-α、IL-1α的表达,可减轻OA关节软骨的损害,IBP、IN在不同时期对软骨细胞IL-1β、TNF-α、IL-1α表达的影响有所不同,其总体抑制效应不明显。     

脂联素抑制人髓核细胞分泌疼痛介质PGE2的初步机制研究

目的验证脂联素在椎间盘源性腰痛患者中可能发挥的抗炎作用,并初步研究其机制。方法首先检测髓核组织内脂联素受体表达水平;随后提取并培养人髓核细胞,检测炎症因子TNF-α对髓核细胞内脂联素受体调控;最后采用慢病毒基因沉默的手段检测脂联素抑制炎症因子诱导髓核细胞产生PGE-2 的作用及机制。结果椎间盘源性腰痛患者髓核组织内脂联素受体Adipo R1/2 表达水平低于正常髓核组织（p <0.05）,炎症介质TNF-α能抑制脂联素受体AdipoR1/2 的表达（p <0.05）。当脂联素处理髓核细胞后,TNF-α对PGE-2 分泌的诱导受抑并且呈浓度剂量依赖效应（p <0.05）;髓核细胞过表达Adipo R1/R2 后,均能抑制炎症因子TNF-α诱导髓核细胞产生PGE-2,其中以Adipo R2 为明显（p <0.05）。结论脂联素能够通过两个受体Adipo R1、Adipo R2 抑制抑制髓核细胞合成分泌疼痛介质PGE-2,提示脂联素可能为一个保护性因子在椎间盘源性腰痛环境中发挥抗炎作用。     

p53半剂量缺失纠正1，25（OH）2D3缺乏小鼠的骨质疏松

目的：探索p53半剂量缺失杂合子小鼠能否通过增强抗氧化能力纠正活性维生素D（1,25（OH）₂D₃）缺乏引起的骨质疏松。方法：取10周龄高钙高磷饮食喂养的同窝野生型（wild type,WT）小鼠、p53半剂量缺失杂合子（p53^+/-）小鼠、1α-羟化酶基因敲除［1α（OH）ase^-/-］小鼠及p53半剂量缺失的1α羟化酶基因敲除［1α（OH）ase^-/-p53^+/-］小鼠的长骨,利用X线、micro-CT、组织病理学和分子生物学等方法,比较各组小鼠血清学、长骨骨矿化、骨形成、骨吸收以及氧化应激等表达变化。结果：与WT小鼠相比,p53^+/-小鼠血清钙、磷、甲状旁腺素（parathyroid hormone,PTH）和1,25（OH）₂D₃水平差异无统计学意义,骨密度、总胶原 （total collagen,T-col）阳性面积、成骨细胞数量、碱性磷酸酶（alkaline phosphatase,ALP）和Ⅰ型胶原（collagen typeⅠ,Col-Ⅰ）阳性面积均有所增加,活性氧水平降低,抗氧化酶SOD1表达增加。与1α（OH）ase^-/-小鼠相比,1α（OH）ase^-/-p53^+/-小鼠血清钙、磷和PTH差异无统计学意义,血清中检测不到1,25（OH）₂D₃,骨密度、T-col阳性面积、成骨细胞数量、ALP和Col-Ⅰ阳性面积均明显增加,破骨细胞数量减少,活性氧水平降低,抗氧化酶SOD1表达增加。结论：p53半剂量缺失可通过增强抗氧化能力纠正 1,25（OH）₂D₃缺乏小鼠的骨质疏松。     

大骨节病、骨关节炎软骨细胞分泌IL-1β、TNF-α及透明质酸对其影响实验研究

目的:探讨在不同剂量透明质酸作用下,人大骨节病(KBD)软骨细胞和骨关节炎(OA)软骨细胞对IL-1β、TNF-α合成分泌的影响,为临床上透明质酸钠治疗OA及KBD提供实验依据。方法:将符合临床诊断标准的KBD及OA患者与从遭遇意外事故的病人作为正常对照,分离、体外培养三组患者关节软骨细胞,分别给予100μg/ml(H100组)、500μg/ml(H500组)不同剂量的透明质酸钠(HA)干预及阴性对照(0.00μg/ml,H0组),通过放免测定细胞培养上清液中IL-1β、TNF-α的含量变化。结果:与正常组比较,KBD和OA组软骨细胞IL-1β、TNF-α明显增高;HA干预随其剂量增高抑制IL-1β、TNF-α合成增强,对TNF-α的抑制有统计学意义。结论:骨关节炎和大骨节病患者软骨细胞合成分泌IL-1β、TNF-α明显增加;应用透明质酸后,可降低两种炎性因子的分泌。     

肿瘤坏死因子-α对软骨细胞蛋白聚糖代谢的影响及小白菊内酯的保护作用

目的 探讨小白菊内酯(PAR)对关节软骨细胞蛋白聚糖合成的调节作用.方法 软骨细胞取自骨关节炎(OA)患者进行膝关节置换术的股骨髁,随机分为4组:对照组、肿瘤坏死因子-α(TNF-α)组、PAR组和TNF-α+PAR组.用3种方法测定蛋白聚糖的代谢:(1)测定蛋白聚糖的代谢产物糖胺多糖(GAG)含量.(2)用针对蛋白聚糖单克隆抗体(Mab-383和5D4),采用ELISA法检测培养液中蛋白聚糖的代谢片段含量.(3)半定量RT-PCR检测软骨细胞蛋白聚糖、ADAMTS-4、ADAMTS-5、基质金属蛋白酶组织抑制因子-1(TIMP-1)和丝裂原活化蛋白激酶.1(MAPK-1)的mRNA 表达.结果 (1)TNF-α+PAR组培养液中GAG的百分含量明显低于TNF-α组(t=5.92,P=0.02),其降低程度与PAR呈剂量依赖性.(2)TNF-α组培养液中5D4片段的含量明显高于TNF-α+PAR组(t=7.93,P=0.016),而各组间培养液中383片段的含量无差别.(3)TNF-α组的蛋白聚糖mRNA表达明显低于其他3组(F=133.7,P=0.000);而其ADAMTS-5及MAPK-1的mRNA表达明显高于其他3组(F=209.7,117.1;P=0.000).结论 (1)TNF-α可促进人OA关节软骨细胞蛋白聚糖的降解而PAR可抑制其降解作用.(2)PAR可下调TNF-α导致的ADAMTS-5及MAPK-1的mRNA表达.     

阿法骨化醇对新诊断2型糖尿病患者胰岛β细胞第一时相分泌的影响

目的通过观察阿法骨化醇[1α-hydroxyvitamin D3,1,α-(OH)2D3]对初诊2型糖尿病患者胰岛β细胞第一时相分泌及血浆中25羟维生D[25-hydroxy vitamin D,25(OH)D]水平的影响,旨在初步探讨它们之间的相互关系。方法受试者分3组:初诊2型糖尿病组(Type 2 Diabetes mellitus,T2DM组)158例、初诊2型糖尿病+阿法骨化醇治疗组[T2DM+1,α-(OH)2D3组]155例及正常对照组(NGT组)152例,以葡萄糖刺激后3到5min急性胰岛素反应(3-5minutes acute insulin response,AIR3-5)来表示第一时相胰岛素分泌功能指数,测定胰岛β细胞第一时相胰岛素分泌功能指数、血浆中25(OH)D及血脂水平并进行比较分析。结果 T2DM组TC、TG、LDL-C、BMI均高于对照组,而AIR3-5、25(OH)D、HDL-C低于对照组,差异具有统计学意义(P<0.05);T2DM+1,α-(OH)2D3组AIR 3-5、25(OH)D、TG、HDL-C与正常对照组比较,差异无统计学意义(P>0.05),而TC、TG、LDL-C、BMI均高于对照组,差异具有统计学意义(P<0.05);T2DM+1,α-(OH)2D3组AIR 3-5、25(OH)D、HDL-C明显高于T2DM组,差异具有统计学意义(P<0.05),而TG明显低于T2DM组,差异具有统计学意义(P<0.05),TC、LDL-C、BMI与T2DM组比较,差异无统计学意义(P>0.05)。AIR 3-5与25(OH)D、HDL-C呈正相关,与TC、TG、LDL-C、BMI均呈负相关。结论初诊T2DM患者25(OH)D、AIR 3-5水平降低,而使用阿法骨化醇治疗组,血浆中25(OH)D水平升高,胰岛β细胞第一时相胰岛素分泌功能得以明显改善,因此,阿法骨化醇提高了血浆中25(OH)D水平,改善了胰岛β细胞第一时相胰岛素分泌功能,同时改善了脂质代谢,可能对防治糖尿病及改善脂质代谢具有重要意义。     

TNF-α诱导大鼠软骨细胞焦亡模型的建立与评价

目的 建立肿瘤坏死因子-α(TNF-α)诱导的大鼠软骨细胞焦亡模型并评价模型的特点。方法 两步酶消化法获取大鼠膝关节软骨细胞,倒置相差显微镜观察大鼠软骨细胞的形态结构,甲苯胺蓝染色、Ⅱ型胶原(ColⅡ)免疫组化染色鉴定大鼠软骨细胞,用不同质量浓度TNF-α(5、10、20、40 ng/ml)诱导建立大鼠软骨细胞的焦亡模型,以TNF-α(0 ng/ml)为对照组。CCK-8法检测大鼠软骨细胞活力,Western blot检测大鼠软骨细胞焦亡信号相关蛋白,确定最佳刺激浓度,ELISA法检测细胞培养上清液中IL-1β、IL-18的水平,免疫荧光检测大鼠软骨细胞gasdermin D(GSDMD)表达,扫描电镜观测大鼠软骨细胞焦亡的形态学改变。结果 与对照组相比,大鼠软骨细胞的活力随着TNF-α浓度升高而逐渐下降,并且核苷酸结合寡聚化结构域样受体蛋白3 (NLRP3)、半胱天冬酶1(caspase-1)、GSDMD、磷酸化核转录因子-κB p65(P-p65)蛋白的表达增加。TNF-α(20 ng/ml)可诱导大鼠软骨细胞基质金属蛋白酶-13(MMP-13)表达上调,ColⅡ表达减少,GSDM...     

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…     

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.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…