低氧/低氧诱导因子-1α通路对成骨细胞与破骨细胞耦联的调控作用

目的 探讨低氧/低氧诱导因子(HIF) -1α通路对成骨细胞与破骨细胞耦联的调控作用.方法 取出生2～3d条件性基因敲除小鼠颅盖骨的成骨细胞与4～8周龄C57BL/6小鼠股骨破骨细胞的前体细胞,建立基因敲除小鼠成骨细胞与破骨细胞前体细胞共培养体系(野生型、HIF-1α-/-、Vhl-/-、HIF - 1α-/-/Vhl -/-共培养).采用RT-PCR技术检测成骨细胞中核因子κB受体活化因子配体(RANKL) mRNA和骨保护素(OPG) mRNA的表达以及破骨细胞中标志酶基因TRAP mRNA的表达,甲苯胺蓝染色观察破骨细胞溶骨形成的骨陷窝.结果 RT-PCR检测结果显示:与野生型共培养比较,HIF-1α-/-共培养的成骨细胞中RANKL mRNA表达上调、OPG mRNA表达下调(均P＜0.05),破骨细胞TRAP mRNA表达上调;Vhl-/-共培养和HIF-1α-/-/Vhl-/-共培养的成骨细胞中RANKL mRNA表达下调、OPG mRNA表达显著上调(均P＜0.05),破骨细胞TRAP mRNA表达下调;随着共培养时间的延长,成骨细胞中RANKL mRNA和OPG mRNA表达均逐渐减少,而破骨细胞TRAP mRNA表达均逐渐增加.甲苯胺蓝染色倒置显微镜观察显示:共培养第9天,破骨细胞骨吸收陷窝出现;随着共培养时间的延长,骨吸收陷窝面积和深度逐渐增加;与野生型共培养比较,HIF-1α-/-共培养的破骨细胞骨吸收陷窝较大且较深,Vhl-/-和HIF-1α-/-/Vhl-/-共培养的破骨细胞骨吸收陷窝较小且较浅.结论 低氧/HIF-1α通路激活后,成骨细胞抑制破骨细胞的分化功能;低氧/HIF-1α通路阻断后,成骨细胞促进破骨细胞的分化功能.     

骨灵片对骨质疏松大鼠血清OPG/RANKL、PICP和相关细胞因子的影响

目的观察骨灵片对骨质疏松症（OP）大鼠血清OPG、RANKL及PICP和相关细胞因子（IL-6、TNF-α和IL-4）表达的影响。方法随机将大鼠分为正常对照组、模型组、骨灵片高、中、低剂量组,采用去势法建立大鼠OP动物模型,术后1个月,骨灵片组给予骨灵片,给药后3个月,采用ELISA法检测血清OPG、RANKL及PICP和IL-6、TNF-α、IL-4。结果与模型组比较,骨灵片各组的OPG、PICP值显著升高（P〈0.05,P〈0.01）,而RANKL、IL-6和TNF-α值均显著下降（P〈0.01,P〈0.01,P〈0.05）,骨灵片高、中剂量组的IL-4值显著升高（P〈0.05）。结论骨灵片可能通过上调OPG,下调RANKL的表达,影响OPG系统,并可能通过调节IL-6、TNF-α、IL-4水平,影响破骨细胞功能,抑制骨吸收,同时可能通过增加PICP表达而使I型胶原的合成增加,进而影响骨的韧性和强度,达到防治OP的目的。     

共培养体系中重组结核杆菌热休克蛋白10对破骨细胞相关基因表达的影响

目的观察重组结核杆菌热休克蛋白10（CPN10）对成骨细胞（OB）-外周血单个核细胞（PBMs）共培养体系中破骨细胞生成及相关基因表达的影响。方法建立培养上清相通但二者互相不接触的成骨细胞一单个核细胞共育模型。实验分对照组和CPN10（10μg/ml）处理组。主要观察指标：①采用TRAP染色及扫描电镜检测破骨细胞生成及小牛骨磨片吸收陷窝,②应用Realtime PCR检测与破骨细胞生成相关基因NFATc1、c-Fos、RANKL、OPG的基因表达。结果两组细胞均有TRAP阳性多核破骨细胞生成,并在小牛骨磨片上形成吸收陷窝;但对照组所获TRAP阳性多核细胞数目、吸收陷窝数目及面积均显著小于CPN10组。Realtime PCR检测结果显示CPN10组与对照组相比NFATc1、c-Fos、RANKL、OPG 相对浓度分别为33.4798±177;2.0929、47.974±177;5.1628、47.0861±177;2.2033、7.4642±177;0.6791（P〈0.05）,对照组各基因表达均显著低于CPN10组。结论 CPN10在成骨细胞-单个核细胞（OB-PBMs）共培养体系中可促进OC的生成及骨吸收,CPN10通过对成骨细胞的作用,致其分泌的OPG/RANKL比例失调,并上调破骨细胞相关基因NFATc1、c-Fos、RANKL、OPG的基因表达。     

TNF-α对人牙周膜成纤维细胞RANKL/OPG mRNA表达的影响

目的 探讨肿瘤坏死因子-α（TNF-α）对人牙周膜成纤维细胞（HPDLFs）的核因子-κB受体活化因子配体（RANKL）及其饵受体骨保护素（OPG）mRNA表达的影响,了解TNF-α在破骨细胞性骨吸收调节中的作用机制。方法 将体外培养的人牙周膜成纤维细胞用不同浓度的TNF-α处理24h,通过半定量逆转录聚合酶链反应（RT-PCR）来观察人牙周膜成纤维细胞OPG、RANKL mRNA表达强度变化。结果 TNF-α上调人牙周膜成纤维细胞的RANKLmRNA表达,RANKL／OPG比率呈上升趋势。结论TNF-α通过调节人牙周膜成纤维细胞的RANKL／OPG的比值可间接地调节破骨细胞分化和活性,从而影响骨代谢。     

口腔鳞状细胞癌侵犯颌骨与破骨细胞及相关细胞因子的关系

目的:研究在口腔鳞状细胞癌侵犯颌骨的病例中,破骨细胞及与破骨细胞相关的两个重要[细胞因子细胞核因子κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)和骨保护素(osteoprotegerin,OPG)]的表达情况.方法:选用12例口腔鳞状细胞癌侵犯颌骨病例的手术标本,将新鲜软组织标本制作成冰冻切片,含骨组织标本经固定脱钙后制成石蜡切片.分别对两种切片进行抗酒石酸酸性磷酸酶(TRAP)染色,以及针对RANKL和OPG两种细胞因子的免疫组织化学染色.结果:TRAP阳性的多核细胞出现在肿瘤与颌骨的交界面附近.其下方的骨组织成锯齿样,TRAP阳性细胞位于骨表面的陷窝内.RANKL免疫组化染色阳性的部位主要有血管内皮和上皮基底膜,在与骨面相近的肿瘤组织中也可见位于血管周围的染色阳性细胞,而在靠近骨面的鳞状细胞癌周围的组织内未见OPG特异性染色阳性.结论:在口腔鳞状细胞癌引起颌骨侵犯的病例中,破骨细胞是受到鳞状细胞癌的影响而分化和活化,并最终引起骨组织的吸收破坏.肿瘤细胞是跟随在破骨细胞后面侵入骨组织,并不直接引起骨的变化.骨侵犯性鳞状细胞癌的细胞及周围的血管可以释放出较多的RANKL类细胞因子,促进破骨细胞的分化和活化,最终导致了破骨活动的发生和发展.     

肿瘤坏死因子-α诱导外周血单核细胞分化为破骨细胞

目的:验证肿瘤坏死因子-α(TNF-α)是否可以直接诱外周血单核细胞(PBMCs)分化为具有骨吸收作用的破骨细胞. 方法:小白鼠PBMCs体外培养中加入TNF-α和白细胞介素-1α(IL-1α)及巨噬细胞克隆集落刺激因子(M-CSF);同时,分别加入细胞核因子κB受体活化因子配体(RANKL)的拮抗剂RANK:Fc和抗TNF-α中和抗体. 对培养终末细胞作组织化学染色,检测破骨细胞特征性标志物抗酒石酸磷酸酶(TRAP)的表达;并以象牙磨片上虫蚀样骨吸收陷窝的形成为指标检测其生物学活性. 结果:PBMCs体外培养形成大量TRAP阳性的多核细胞(MNCs);象牙磨片上见到虫蚀样骨吸收陷窝,后者的面积对TNF-α呈剂量依赖性. RANK:Fc对此现象无抑制作用,而抗TNF-α中和抗体可完全阻断该现象的发生. 结论:TNF-α能够直接诱导PBMCs分化为具有骨吸收功能的破骨细胞.     

重组人骨形态发生蛋白2缓释体对铬磨损颗粒诱导的溶骨效应的影响

摘要：目的建立大鼠植骨气囊模型,观察在不同浓度重组人骨形态发生蛋白2（rhBMP-2）缓释体干预下气囊内组织的破骨细胞
分化因子（RANKL）、骨破坏素（OPG）表达情况。方法在大鼠背部注入空气形成气囊,取同源大鼠的颅骨植入气囊内。将已制
成的植骨气囊模型大鼠分成5组：空白组、铬颗粒组、50 μg/L rhBMP-2缓释体+铬颗粒组、100 μg/L rhBMP-2缓释体+铬颗粒组
和200 μg/L rhBMP-2缓释体+铬颗粒组。各组分别用药处理,2周后取出囊腔内组织进行RANKL、OPG的Wester-blot、Rt-PCR
检测,并对囊腔内骨片行TRAP染色,用计算机图像分析技术测定骨片破骨细胞染色面积,进行药效评价。结果RANKL、OPG
在各组囊腔组织中均有表达。骨片破骨细胞染色面积、RANKL、OPG的Wester-blot和RT-PCR结果及RANKL/OPG值,在铬颗
粒组中明显高于其他4 组(P<0.05);在3个rhBMP-2缓释体干预组内,随着rhBMP-2缓释体浓度增高呈递减趋势,组间差异均有
统计学意义(P<0.05);在空白组和200 μg/L rhBMP-2缓释体+铬颗粒组间未见明显差异（P>0.05）。结论大鼠植骨气囊模型成
功的模拟了人工关节无菌性松动的生物学和组织学特征。铬颗粒可以引起RANKL/OPG值升高,促进溶骨;rhBMP-2缓释体
通过降低RANKL/OPG值,从而促进成骨、抑制骨吸收效益。
     

成纤维母细胞样基质细胞促进假体周围骨溶解

目的 探讨松动假体周围的假膜组织中成纤维母细胞样基质细胞在假体周围骨溶解中的作用.方法 从人工假体周围的假膜标本中分离成纤维母细胞样基质细胞并作体外培养,传代后与外周血单核细胞共培养,并向各组内分别加入骨水泥颗粒和破骨细胞形成抑制因子(OPG)等,检测是否有破骨细胞形成及其活性;并采用半定量RT-PCR法检测成纤维母细胞样基质细胞中核因子κ B受体活化因子配基(RANKL)和OPG等破骨细胞调节因子mRNA表达量的变化.结果 细胞共培养结束时观察到抗酒石酸磷酸酶(TRAP)和玻璃蛋白酶受体(VNR)阳性的多核细胞及骨吸收陷窝形成,这一过程可被OPG彻底阻断;骨水泥颗粒组骨吸收陷窝的数量较对照组增加.同时基质细胞表达RANKL和OPG mRNA,骨水泥颗粒组的表达量较对照组分别增加,并且RANKL/OPG的比率增大.结论 人工假体周围假膜中成纤维母细胞样基质细胞表达膜蛋白型及可溶性RANKL,支持破骨细胞分化及其骨吸收活性,从而促进假体周围骨溶解.     

抗坏血酸抑制核因子κB受体活化子配体诱导RAW264.7细胞的分化和功能

目的研究抗坏血酸对核因子κB受体活化子配体(RANKL)诱导体外培养的破骨前体细胞RAW2647形成成熟多核破骨细胞的影响,探讨抗坏血酸在破骨前体细胞分化中的作用及机制。方法利用不同浓度的抗坏血酸与RANKL单独或共同处理RAW2647细胞,MTT法测定细胞增殖,抗酒石酸酸性磷酸酶(TRAP)染色法观察TRAP阳性多核细胞,逆转录聚合酶链反应(RTPCR)测定破骨细胞表型基因和功能基因的表达,Western印迹方法检测碳酐酶Ⅱ基因蛋白表达,破骨细胞的骨吸收功能用骨吸收陷窝面积计数法分析。结果抗坏血酸和RANKL都抑制RAW2647细胞的增殖(P<005),但它们之间无协同作用。抗坏血酸本身不能诱导RAW2647细胞形成破骨细胞,但可抑制RANKL诱导的TRAP阳性多核破骨细胞形成(P<005)。抗坏血酸下调RANKL诱导的碳酐酶Ⅱ和RANK基因mRNA表达及碳酐酶Ⅱ基因蛋白表达,抑制骨吸收功能(P<005)。结论抗坏血酸能直接抑制RANKL诱导的破骨细胞形成和功能。     

白芦藜醇对大鼠牙移植后牙周组织愈合及Sirt1/Nrf2信号通路的影响

目的：观察白芦藜醇（RSV）对大鼠牙移植后牙周组织愈合及Sirt1/Nrf2信号通路的影响。方法：48只雄性Lewis大鼠随机分为对照（Control）组、模型（Model）组、低剂量RSV(L-RSV)组和高剂量RSV(H-RSV)组。苏木精-伊红（HE）染色、抗酒石酸酸性磷酸酶（TRAP）染色观察牙周组织病理学变化;免疫组织化学染色检测核因子κB受体激活剂配体（RANKL）、骨保护素（OPG）、Runt相关转录因子2(RUNX2)和骨钙素（OCN）等成骨分化相关因子的表达;扫描电子显微镜分析牙根吸收情况,Western blot检测牙周组织中Sirt1、Nrf2蛋白表达水平。结果：与Control组比较,Model组牙周组织TRAP阳性细胞数量增多,RANKL表达和牙根吸收率升高,OPG、RUNX2和OCN表达以及Sirt1、Nrf2蛋白水平降低（P<0.05）。与Model组比较,L-RSV组和H-RSV组牙周组织TRAP阳性细胞数量减少;RANKL表达和牙根吸收率降低,OPG、RUNX2和OCN表达以及Sirt1、Nrf2蛋白水平升高（P<0.05）。结论：RSV可...     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.