Effects of applied DC electric field on ligament fibroblast migration and wound healing

Applied electric fields (static and pulsing) are widely used in orthopedic practices to treat nonunions and spine fusions and have been shown to improve ligament healing in vivo. Few studies, however, have addressed the effect of electric fields (EFs) on ligament fibroblast migration and biosynthesis. In the current study, we applied static and pulsing direct current (DC) EFs to calf anterior cruciate ligament (ACL) fibroblasts. ACL fibroblasts demonstrated enhanced migration speed and perpendicular alignment to the applied EFs. The motility of ligament fibroblasts was further modulated on type I collagen. In addition, type I collagen expression increased in ACL fibroblasts after exposure to pulsing EFs. In vitro wound-healing studies showed inhibitory effects of static EFs, which were alleviated with a pulsing EF. Our results demonstrate that applied EFs augment ACL fibroblast migration and biosynthesis and provide potential mechanisms by which EFs may be used for enhancing ligament healing and repair.     

Gating of the HypoPP-1 mutations: I. Mutant-specific effects and cooperativity

Hypokalemic periodic paralysis type 1 (HypoPP-1) is a hereditary muscular disorder caused by point mutations in the gene encoding the voltage-gated Ca²⁺ channel α subunit (Ca_v1.1). Despite extensive research, the results on HypoPP-1 mutations are minor and controversial, as it is difficult to analyse Ca²⁺ channel activation macroscopically due to an existence of two open states. In this study, we heterologously expressed the wild-type and HypoPP-1 mutations introduced into the rabbit cardiac Ca²⁺ channel (R650H, R1362H, R1362G) in HEK-293 cells. To examine the cooperative effects of the mutations on channel gating, we expressed two double mutants (R650H/R1362H, R650H/R1362G). We performed whole-cell patch-clamp and, to obtain more information, applied a global fitting procedure whereby several current traces elicited by different potentials were simultaneously fit to the kinetic model containing four closed, two open and two inactivated states. We found that all HypoPP-1 mutations have “loss-of-function” features: D4/S4 mutations shift the equilibrium to the closed states, which results in reduced open probability, shorter openings and, therefore, in smaller currents, and the D2/S4 mutant slows the activation. In addition, HypoPP-1 histidine mutants favored the second open state O₂ with a possibly lower channel selectivity. Cooperativity between the D2/S4 and D4/S4 HypoPP-1 mutations manifested in dominant effects of the D4/S4 mutations on kinetics of the double mutants, suggesting different roles of D2/S4 and D4/S4 voltage sensors in the gating of voltage-gated calcium channels. Alexey Kuzmenkin and Chao Hang contributed equally to this work.     

PVC/PE/PVDC固体药用复合硬片PVDC涂布量测定

目的 建立一种测定聚氯乙烯/聚乙烯/聚偏二氯乙烯（PVC/PE/PVDC）固体药用复合硬片PVDC涂布量的方法。方法 裁取10 cm×10 cm的样品5片,将样品先在丙酮中浸泡,分离得到PVC层和PE/PVDC复合层。将PE/PVDC复合层于80℃中干燥2 h后,于23℃放置4 h,精密称定。再将PE/PVDC复合层在四氢呋喃中浸泡,使PVDC层溶解在四氢呋喃溶液中,得到PE膜层,于80℃中干燥2 h后,于23℃放置4 h,精密称定,前后2次差值即为PVDV质量（g）。最后计算PVDC涂布量（以g·m^-2表示）。结果 5批样品PVDC涂布量测定结果与标示涂布量均一致,且偏差均在±5%之内。结论 该方法准确、灵敏,适用于PVC/PE/PVDC固体药用复合硬片PVDC涂布量的测定。     

<Emphasis Type="Italic">In Vitro</Emphasis>-<Emphasis Type="Italic">In Vivo</Emphasis> Dose Response of Ursolic Acid,Sulforaphane, PEITC,and Curcumin in Cancer Prevention

According to the National Center of Health Statistics, cancer was the culprit of nearly 600,000 deaths in 2016 in the USA. It is by far one of the most heterogeneous diseases to treat. Treatment for metastasized cancers remains a challenge despite modern diagnostics and treatment regimens. For this reason, alternative approaches are needed. Chemoprevention using dietary phytochemicals such as triterpenoids, isothiocyanates, and curcumin in the prevention of initiation and/or progression of cancer poses a promising alternative strategy. However, significant challenges exist in the extrapolation of in vitro cell culture data to in vivo efficacy in animal models and to humans. In this review, the dose at which these phytochemicals elicit a response in vitro and in vivo of a multitude of cellular signaling pathways will be reviewed highlighting Nrf2-mediated antioxidative stress, anti-inflammation, epigenetics, cytoprotection, differentiation, and growth inhibition. The in vitro-in vivo dose response of phytochemicals can vary due, in part, to the cell line/animal model used, the assay system of the biomarker used for the readout, chemical structure of the functional analog of the phytochemical, and the source of compounds used for the treatment study. While the dose response varies across different experimental designs, the chemopreventive efficacy appears to remain and demonstrate the therapeutic potential of triterpenoids, isothiocyanates, and curcumin in cancer prevention and in health in general.     

In vitro activities of various antimicrobials alone and in combination with tigecycline against carbapenem-intermediate or -resistant Acinetobacter baumannii

The activities of tigecycline alone and in combination with other antimicrobials are not well defined for carbapenem-intermediate or -resistant Acinetobacter baumannii (CIRA). Pharmacodynamic activity is even less well defined when clinically achievable serum concentrations are considered. Antimicrobial susceptibility testing of clinical CIRA isolates from 2001 to 2005 was performed by broth or agar dilution, as appropriate. Tigecycline concentrations were serially increased in time-kill studies with a representative of the most prevalent carbapenem-resistant clone (strain AA557; imipenem MIC, 64 mg/liter). The in vitro susceptibility of the strain was tested by time-kill studies in duplicate against the average free serum steady-state concentrations of tigecycline alone and in combination with various antimicrobials. Ninety-three CIRA isolates were tested and were found to have the following antimicrobial susceptibility profiles: tigecycline, MIC(50) of 1 mg/liter and MIC(90) of 2 mg/liter; minocycline, MIC(50) of 0.5 mg/liter and MIC(90) of 8 mg/liter; doxycycline, MIC(50) of 2 mg/liter and MIC(90) of > or =32 mg/liter; ampicillin-sulbactam, MIC(50) of 48 mg/liter and MIC(90) of 96 mg/liter; ciprofloxacin, MIC(50) of > or =16 mg/liter and MIC(90) of > or =16 mg/liter; rifampin, MIC(50) of 4 mg/liter and MIC(90) of 8 mg/liter; polymyxin B, MIC(50) of 1 mg/liter and MIC(90) of 1 mg/liter; amikacin, MIC(50) of 32 mg/liter and MIC(90) of > or =32 mg/liter; meropenem, MIC(50) of 16 mg/liter and MIC(90) of > or =128 mg/liter; and imipenem, MIC(50) of 4 mg/liter and MIC(90) of 64 mg/liter. Among the tetracyclines, the isolates were more susceptible to tigecycline than minocycline and doxycycline, according to FDA breakpoints (95%, 88%, and 71% of the isolates were susceptible to tigecycline, minocycline, and doxycycline, respectively). Concentration escalation studies with tigecycline revealed a maximal killing effect near the MIC, with no additional extent or rate of killing at concentrations 2x to 4x the MIC for tigecycline. Time-kill studies demonstrated indifference for tigecycline in combination with the antimicrobials tested. Polymyxin B, minocycline, and tigecycline are the most active antimicrobials in vitro against CIRA. Concentration escalation studies demonstrate that tigecycline may need to approach concentrations higher than those currently achieved in the bloodstream to adequately treat CIRA bloodstream infections. Future studies should evaluate these findings in vivo.     

维兰特罗/乌美溴铵联合罗氟司特对COPD患者肺功能和mMRC评分的影响

目的研究维兰特罗/乌美溴铵联合罗氟司特对慢性阻塞性肺病(COPD)患者肺功能和改良英国医学研究组理事会呼吸困难指数(mMRC) 评分的影响。方法COPD患者120例随机分为观察组以及对照组各60例,均采用维兰特罗/乌美溴铵进行治疗,观察组在此基础上联合采用罗氟司特进行治疗,比较两组治疗效果、肺功能、mMRC评分、免疫功能及不良反应。结果观察组治疗总有效率高于对照组(P＜0.05);治疗后两组用力呼气量(FEV)、第1秒用力呼气量(FEV1)、第1秒用力呼气量占用力肺活量百分率(FEV1%)升高,且观察组高于对照组(P＜0.05);观察组mMRC评分低于对照组(P＜0.05);治疗后两组CD3＋、CD4＋、CD8＋、CD4＋/CD8＋水平显著升高,且观察组高于对照组(P＜0.05)。结论维兰特罗/乌美溴铵联合罗氟司特治疗COPD患者其肺功能以及mMRC评分显著改善,建议临床推广。     

miR-155靶向调节PIK3R1对类风湿关节炎大鼠模型PI3K/Akt/mTOR信号通路的影响

目的 探讨miR-155靶向调节PIK3R1对类风湿关节炎（rheumatoid arthritis,RA）大鼠PI3K/Akt/mTOR信号通路的影响。方法 SD大鼠随机分为对照组、模型组、miR-155 agomir组、miR-155 antagomir组、miR-155阴性对照组,诱导RA模型,分组处理后,观察关节症状,检测关节炎指数及后足趾容积;HE染色观察大鼠关节组织病理形态;使用试剂盒检测大鼠关节组织炎性因子IL-6、IL-17及IL-18水平;免疫印迹检测大鼠关节组织PI3K/Akt/mTOR信号通路蛋白表达;qRT-PCR实验检测大鼠关节组织miR-155及PIK3R1 mRNA水平;双荧光素酶报告基因实验检测miR-155对PIK3R1的靶向调控作用。结果 与对照组比较,模型组大鼠关节炎症状明显,关节炎指数、后足趾容积、关节组织炎性因子IL-6、IL-17及IL-18水平、关节组织p-PI3K/PI3K、p-Akt/Akt及p-mTOR/mTOR水平、关节组织miR-155水平明显升高（P＜0.05）,PIK3R1 mRNA水平明显降低（P＜0.05）。与模型组、miR-155阴性对照组比较,miR-155 antagomir组大鼠上述指标得到明显改善（P＜0.05）;miR-155 agomir组与miR-155 antagomir组趋势相反（P＜0.05）。结论 miR-155可靶向下调PIK3R1的表达,激活PI3K/Akt/mTOR信号,加重RA大鼠关节炎症损伤,下调miR-155表达,可抑制PI3K/Akt/mTOR信号激活及炎症反应发生发展,改善关节炎症状。     

Understanding Racially Diverse Community Member Views of Obesity Stigma and Bariatric Surgery

Obesity Surgery - The obesity epidemic poses serious challenges to health equity. Despite bariatric surgery being one of the most effective obesity treatments, utilization remains low. In this...