目的:建立一种降低HPLC-ELSD国产流动相试剂(甲醇、乙腈、二氯甲烷等)噪音的方法.方法:将流动相试剂置蒸馏装置中进行常压蒸馏、水浴加热,水温控制在高出被蒸馏试剂沸点5-20℃,再将馏出液经有机滤膜过滤、超声波脱气.结果:经本法处理后的国产流动相试剂ELSD噪音得到明显降低,达到正常检测的要求.结论:该方法简单、有效、可明显降低HPLC-ELSD国产流动相试剂的噪音.

背景:前期研究结果显示,透骨消痛胶囊能降低软骨细胞炎症因子表达,减少软骨细胞凋亡,促进软骨细胞增生,延缓软骨、软骨下骨退变,从而缓解骨关节炎疾病进程,减轻炎症反应,但该方对滑膜病变的作用机制尚不明确.目的:从早期骨关节炎滑膜炎症水肿与相关水通道蛋白调节的角度,进一步探讨透骨消痛胶囊抗炎消肿的作用机制.方法:24只SD大鼠随机分成空白组、模型组、中药组3组,后两组应用木瓜蛋白酶法制备早期骨关节炎模型1周后,分别以透骨消痛胶囊(中药组)或生理盐水(空白组和模型组)灌胃4周.小动物MRI观察膝关节形态结构,放射免疫法测定血清和关节液白细胞介素1β和肿瘤坏死因子α水平,石蜡切片观察滑膜组织显微结构,透射电镜观察滑膜细胞超微结构,免疫组化/荧光染色定量、定位水通道蛋白1和水通道蛋白3的表达.实验方案经福建中医药大学动物实验伦理委员会批准(批准号为[2019]福中医伦理审字第024号).结果与结论:①与模型组相比,中药组滑膜水肿减轻、滑液分泌减少;血清和关节液白细胞介素1β和肿瘤坏死因子α水平明显下降;滑膜细胞增生减少,组织纤维化和血管增生明显减轻;滑膜细胞粗面内质网、高尔基体、囊泡和溶酶体等减少;水通道蛋白1和水通道蛋白3表达显著下调,散在分布于细胞膜和细胞质,无聚集现象;②结果说明,透骨消痛胶囊可能通过减少滑膜水通道蛋白1和水通道蛋白3含量,减少滑液分泌,改善关节水肿,这可能是该药物抗炎消肿,延缓骨关节炎进展的作用机制之一.

Doxorubicin (Dox), an effective antineoplastic drug, was limited use for cardiotoxicity. Xinshuitong Capsule (XST), a patented herbal formula, showed desirable beneficial effects in the treatment of chronic heart failure (CHF) patients. However, the drug on Dox-induced cardiotoxicity remains unclear. Ninety male Sprague-Dawley rats were randomized into two groups: 15 rats were selected as the normal group and 75 rats were injected intraperitoneally with Dox to establish CHF rat models, the success ones were randomly divided into five groups: low XST (LXST), medium XST (MXST) or high XST (HXST) (4.9, 9.8, or 19.6 g/kg d) administrated intragastrically twice a day for 4 weeks, with the captopril-treated group and the model group as comparison. The model group showed the cardiac functions generally impaired, and CHF mortality rate higher (47%) than those in the XST-treated groups (averaged 24%, P < 0.05). Compared with XST-treated groups, myocardial remodeling, inflammation and desarcomerization, and higher water content more severe in the cardiac tissue in the model group (P < 0.05), which was associated with higher expressions of mRNA or protein levels of AQP1, 4 and 7. Dox-impaired cardiac functions, cardiac remodeling and myocardial edema could be dose-dependently reverted by XST treatment. XST could inhibit AQP1, 4 and 7 at mRNA levels or at protein levels, which was associated with the attenuation of myocardial edema and cardiac remodeling, decreasing the ventricular stiffness and improving the cardiac functions and rats' survival. AQPs is involved in cardiac edema composed one of the mechanisms of Dox-induced cardiotoxicity, XSTvia inhibition of AQPs relieved the Dox-induced side effects.

目的 采用Box-Behnken响应面法优选牛至中百里香酚与香芹酚的提取工艺.方法 在单因素实验的基础上,以百里香酚与香芹酚提取率之和为评价指标,采用Box-Behnken设计法考察乙醇浓度、液固比和药材粒度对提取率的影响.结果 最佳的提取工艺参数为:乙醇浓度体积分数80％,液固比13∶1(mL/g),药材粒度为过40目筛,最高提取率为694.80μg·g-1,与预测值的偏差较小.结论 所优选的提取方法操作简便、成本低、预测性良好,可为牛至中百里香酚与香芹酚的规模化大生产提供参考.

The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg-1·d-1)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.

水通道蛋白(aquaporins,AQPs)是顺渗透压差或浓度梯度进行跨膜水转运的小分子蛋白家族,目前在哺乳动物中发现了13种亚型(AQP0～AQP12).心肌水肿,是多种心脏疾病病理生理过程中常见的病理现象,研究发现心脏AQPs参与其中.应激状态下,心脏AQPs蛋白表达的改变,通过一系列途径影响心肌水液代谢,进一步影响心功能.目前,以肾脏和大脑组织中AQPs研究较多,关于心脏中AQPs的研究较少;心脏中AQP1、AQP4、AQP7较多.因此,本文拟从心脏AQP1、AQP4、AQP7在心肌水肿中的病理生理研究进展进行综述.

The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg-1·d-1)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.

Objective: This study aimed to explore the neuroprotective effect of Baihe Dihuang Tang (BDT), a traditional Chinese herbal decoction used for nervous-mental system diseases, on serum-deprived PC12 cell. Methods: BDT treatment time and concentration were determined through 4,5-dimethylthiazol-2-y1,2,5-diphenyl tetrazolium (MU) assay. PC12 cells were randomly divided into three groups: control (cells cultured in serum-containing medium), model (cells cultured in serum-deprived medium), and BDT (cells cultured in serum-deprived medium and treated with 3 mg/mL BDT for 24 h) groups. Cell morphology was observed under an inverted phase contrast microscope. The ultrastructure of cells was studied under a transmission electron microscopy. Membrane potential was determined using a laser scanning confocal microscope. Total protein and ATP levels were analyzed by the bicinchoninic acid method and firefly luciferase method, respectively. Results: Compared with the model group, PC12 cells in the BDT group exhibited extensive morphology and adhered to the culture plate. The ultrastructure was modified and possessed a smooth membrane and nuclear surface, increased microvilli-like membrane protrusions, homogeneously distributed organelles, rare pinocytosis and phagocytosis, abundant ribosomes, rare vacuoles, and normal-shaped nucleus. BDT also enhanced the total cell protein content, ATP level, and membrane hyperpolarization. Conclusion: BDT could reduce cell death and neural excitability and regulate energy metabolism caused by serum deprivation. We suggest that BDT could modify the hypometabolic state and neural overactivity in chronic fatigue syndrome (CFS). This work may provide new insights into the possibility of using BDT as a therapeutic agent for CFS.

目的 探讨心水通胶囊治疗慢性心力衰竭(CHF)的作用机制. 方法 将48只雄性Wistar大鼠随机分为正常组、模型组、卡托普利组和心水通胶囊(XST)中药水提液低、中、高剂量组,每组8只.除正常组外,其余各组采用阿霉素腹腔注射建立CHF大鼠模型,超声心动图参数左心室射血分数(LVEF) ＜50％则造模成功.卡托普利组按2.625 mg/(kg· d)予卡托普利灌胃;XST低、中、高剂量组分别按4.95、9.9、19.8 g/(kg· d)予XST中药水提液灌胃;正常组和模型组给予等体积蒸馏水.各组均每日灌胃1次,连续干预4周.各组每周称取体质量2次,末次给药后禁食不禁水收集24 h尿液.ELISA法测定各组血清中肾素(Renin)、血管紧张素Ⅱ(AngⅡ)、醛固酮(ADS)水平;全自动生化分析仪检测血清电解质、尿素氮(Cr)、肌酐(BUN)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)水平以及尿液电解质、总蛋白(TP)水平. 结果 模型组血清中Renin、AngⅡ、ADS水平比正常组明显增加(P＜0.01);XST低、中、高剂量组的血清Renin、AngⅡ、ADS水平呈剂量依赖性降低,各组与模型组比较,均有显著差异(P＜0.01).XST各组的Cr、BUN、CK和CK-MB均有不同程度降低,且呈剂量依赖性降低,各组与模型组对照具有显著差异(P＜0.01).同时发现,XST高剂量组和中剂量组的血电解质水平与正常对照组无差异(P＞0.05),XST治疗组大鼠体重比模型组均有不同程度降低,其中,XST高剂量组和中剂量组具有统计学意义,但三个治疗组的利尿效果未见差异. 结论 XST中药水提物通过作用于RAAS,改善心肌代谢及肾功能,维持水电解质平衡,从而起到防治慢性心力衰竭的作用.

BACKGROUND:Inflammatory cytokines enhanced the progress of the pathogenesis of osteoarthritis, however the mechanisms remain unclear. The objective is to determine aquaporins (AQPs) in the pathogenesis of osteoarthritis.METHODS AND FINDINGS:Primary rat articular chondrocytes were treated with IL-1β to mimic the early stage of osteoarthritis in vitro. Early osteoarthritis animal model was established by intra-articular injection of 4% papain. Micro- or ultra-structure histopathologic changes, cell viability, apoptosis cells and cell membrane permeability, locations and expressions of AQP1 and AQP3 and matrix were detected in the cartilage or in the chondrocytes of knee. IL-1β could reduce the chondrocytes viability, increase the apoptosis cells, and also impair the cell membrane and organelles. IL-1β significantly induced the up-regulation of AQP1 and AQP3 in the chondrocytes. In the chondrocytes, AQPs were mainly clustered in both membrane and perinuclear region of cytoplasm, while higher AQPs were detected in the superficial and middle layers of the cartilage. With the up-regulation of AQPs, the cartilage matrix was considerably decreased in both the chondrocytes and in the osteoarthritis cartilage. In the early osteoarthritis rat model, serum and synovial fluid confirmed that higher IL-1β could increase the expressions of AQPs, and decrease the cartilage matrix in both the chondrocytes and the cartilage.CONCLUSIONS:Inflammatory cytokine IL-1β via up-regulation of AQPs caused the abnormal metabolism of water transport and loss of the cartilage matrix in the chondrocytes, and ultimately exacerbated the pathogenesis of early osteoarthritis. Therefore, AQPs may be a candidate therapeutic target for prevention and treatment of osteoarthritis.

目的 研究心水通胶囊水提物(XST)对慢性心衰大鼠N-端脑利钠肽前体(NT-proBNP)、心肌肌钙蛋白(cTNⅠ)、心功能及凝血指标的影响.方法 阿霉素构建慢性心衰大鼠模型,并用不同浓度心水通胶囊水提物干预心衰大鼠.在给药前、给药15 d、给药30 d 3个阶段分别检测每组大鼠心功能.给药30 d后,检测血清N-端脑利钠肽前体、心肌肌钙蛋白、凝血指标浓度,苏木精-伊红(HE)染色观察心肌细胞病理变化.结果 与模型组比较,心水通胶囊水提物中、高剂量组左室收缩末期内径(LVIDs)和左室舒张末期内径(LVIDd)明显降低,射血分数(LVEF)明显升高(P<0.01),血清N-端脑利钠肽前体、心肌肌钙蛋白和纤维蛋白原(FIB)浓度显著降低(P<0.01),凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)和凝血酶时间(TT)水平显著升高(P<0.01),且高剂量组给药30 d后左室收缩末期内径和左室舒张末期内径降低最显著,射血分数升高最显著(P<0.05);同时心水通胶囊水提物能改善心衰大鼠心肌细胞的病理性肥大、变性和坏死.结论 心水通胶囊水提物能剂量依赖性降低心衰大鼠血清N-端脑利钠肽前体、心肌肌钙蛋白浓度,改善高凝血和心功能,提高心衰大鼠的存活率.

The collapse of mitochondrial membrane potential (ΔΨm) resulted in the cell apoptosis and heart failure. Xinshuitong Capsule (XST) could ameliorate left ventricular ejection fraction (LVEF), New York Heart Association (NYHA) classes and the quality of life in patients with chronic heart failure in our clinical study, however, its cardioprotective mechanisms remain unclear.

Survivin is essential to angiogenesis and revascularization, but its role in coronary collateral formation remains unclear. The role of survivin in peripheral blood mononuclear cells (PBMCs) of coronary chronic total occlusion (CTO) patients was investigated. Coronary CTO patients (n=46; mean age 60.1±8.5, male 54.3%) (CTO group) and normal control patients (n=18; mean age 58.0±10.0, male 55.6%) underwent angiographic collateral vessel grading by Rentrop classification (C0 - C3) and provided peripheral blood between June 2006 and February 2007. Rat hind limb ischemia models were constructed using four equal groups of Sprague-Dawley rats (n=36): normal control, sham operation, operation and granulocyte macrophage colony-stimulating factor (GM-CSF). PBMC numbers and characteristics, collateral vessels, survivin, CD4, CD8, CD44, vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1) expression were determined using RT-PCR, flow cytometry, immunocytochemistry and western blot analysis. PBMC survivin mRNA and protein expression levels were higher in patients with good collateral circulation (C2 + C3) than in patients with no collateral flow (C0) (all P<0.05). Survivin single-positive and survivin and CD8, VEGF and ICAM-1 double-positive percentages were elevated in patients with good collateral circulation compared to those with normal and no collateral flow (all P<0.05), consistent with the rat model results, wherein higher survivin levels produced significantly larger and more visible collateral vessels. In conclusion, elevated survivin expression in PBMCs, particularly survivin and CD8, VEGF, and ICAM-1 double-positive PBMCs, may be crucial for good collateral formation in patients with coronary CTO, as confirmed by assessment of a rat model.

OBJECTIVE:To study the purification technology of total flavonoids from Nelumbinis receptaculum by macroporous resin. METHODS:Using adsorption rate and desorption rate of total flavonoids from Nelumbinis receptaculum as index,the type of macroporous resin was selected by static adsorption-desorption tests;With adsorption rate of total flavonoids as index,single factor test was used to investigate the effects of the concentration of total flavonoids,adsorption time,adsorption speed,drug-loading amount,water amount,volume fraction and amount of eluant and other factors on the purification technology. The optimal technology was validated. RESULTS:Among 10 kinds of resin,HPD-400 macroporous resin was found to have the best adsorption and desorption effects. The optimal purification conditions was as follows as the concentration of total flavonoids 7.00 mg/ml,adsorption time of 3 h,flow rate for sampling of 3 column volume(BV)/h,drug-loading amount of 8 BV,water amount of 6BV,50% ethanol elution amount of 4 BV. In validation test,mass fraction of total flavonoids from purified Nelumbinis receptaculum were 63.88%,62.50% and 63.44%(RSD=1.11%,n=3). CONCLUSIONS:HPD-400 macroporous resin could purify total flavonoids from purified Nelumbinis receptaculum,and established purification technology is stable and practical.

Ginseng preparations contain high concentrations of germanium (Ge), which was reported to contribute to diuretic resistance or renal failure. However, Ge content in ginseng and the influence on renal functions remain unclear. Forty rats were randomly divided into control group, low, moderate, and high Ge ginseng-treated group and observed for 25 days. Daily urine, renal functions, and serum and urine electrolytics were measured. Ge retention in the organs and renal histological changes were also evaluated. Ge content ranged from 0.007 to 0.450 µg/g in various ginseng samples. Four groups showed no difference in the daily urine output, glomerular filtration rate, urinary electrolytes excretions, 24 h-urine protein, as well as plasma and urine urea nitrogen, creatinine, osmotic pressure, and pH values. Ge did not cause any renal pathological effects in this study. No Na and water retention was detected in the ginseng-treated groups. Ge retention in various organs was found highest in spleen, followed by the kidney, liver, lung, stomach, heart, and pancreas. The total Ge contents in various ginsengs were low, and ginseng treatment did not affect renal functions or cause renal histological changes.

Aquaporins ( AQPs ) act as an important channel for body water crossing the cell membrane, and are closely related to the balance of water metabolism and water reserves. Recent studies have found that there are signiifcant differences in the expressions of AQPs in arthritis joints and normal joints. The expressions of AQP1, AQP3 and AQP9 in the patients with rheumatoid arthritis and osteoarthritis are all signiifcantly increased, which are enhanced with the occurrence of lesions. The expressions of AQP4 are higher in the patients with gouty arthritis and osteoarthritis than in the patients with normal joints, which can reflect the severity of articular diseases. After medication, the expressions of AQPs are remarkably reduced, and the arthritis symptoms are improved. It points out that a special intimate relationship exists between the upregulation of the expressions of AQPs in arthritis joints and the occurrence and development of pathological changes. With AQPs as therapeutic targets, the cell membrane permeability to water can be reduced through regulation, so as to improve inlfammation. It offers a new research idea for the prevention and control of arthritis. In this paper, the studies on the relationship between AQPs and arthritis in recent years are reviewed.

目的 优化莲房总黄酮的回流提取工艺. 方法 用正交试验进行回流提取莲房总黄酮,以紫外分光光度法测定莲房总黄酮的吸光度,标准曲线法计算莲房总黄酮的提取率. 结果 回流提取莲房总黄酮的影响因素为:提取次数＞乙醇浓度＞料液比＞提取时间;最优提取工艺条件是:提取次数3次,乙醇浓度70％,料液比1∶16,提取时间1.5 h.在此条件下莲房总黄酮的平均提取率为14.33％,RSD为4.03％. 结论 优化后的回流提取工艺提取莲房总黄酮,具有简便、高效、稳定可行的特点.

Objective:To investigate the anti -atherosclerosis mechanism of Glabridin ( GD) whether via the pathway of autophagy.Methods:The cultured human aorta vascular endothelial cells (HAVECs)were pretreated with or without dif-ferent concentrations of GD , then treated with different concentrations of ox -LDL to induce cell injury or autophagy . The detection of cell viability by MTT assay , autophagic bodies by transmission electron microscopy , and autophagy -re-lated protein Beclin1 and LC3 by Western blotting were conducted in different groups .Results:Dose-correlated signifi-cant decrease in cell viability was observed in ox -LDL treated HAVECs than that in blank control group , while the au-tophagic body and the expressions of Beclin 1 and LC3 significantly increased compared with the control group ( P<0.05 ) .By contrast , the HAVEC pretreated by GD exhibited considerable higher in cell viability than the ox -LDL counterparts, while the expressions of autophagic bodies and Beclin 1 and LC3 were significantly lower (P<0.05). Conclusion:The concentration-dependent ox-LDL can induce autophagic death of HAVEC ,and the protective actions of vascular cells or anti -AS effect of GD may be partially via the inhibition of autophagy pathway .