理气中药组方对糖尿病大鼠胃排空延迟的作用

目的:观察理气中药经验组方对糖尿病大鼠胃排空延迟的干预作用。方法:雌雄各半成年SD大鼠110只,随机分为正常组(A组)、糖尿病中药组(B组)、糖尿病胃复安组(C组)、糖尿病组(D组)。A组、D组1次/d按10 mL/只予0.9%生理盐水灌胃,B组1次/d按8 mL/只予中药煎剂灌胃,C组1次/d按0.5 mg/只予胃复安片灌胃。喂养12周后,行13C胃排空实验及甲基橙水溶液胃排空实验,观察各组大鼠胃排空情况。结果:糖尿病大鼠胃排空较正常大鼠明显延迟(P0.01),糖尿病大鼠胃排空延迟模型制作成功。糖尿病中药组和糖尿病胃复安组大鼠胃排空较糖尿病组快(P0.01);糖尿病中药组大鼠胃排空较糖尿病胃复安组大鼠快(P0.05)。结论:常规喂养12周后糖尿病大鼠出现胃排空延迟。理气中药陈皮、枳实、木香、香附组方煎剂和胃复安均能促进糖尿病大鼠胃排空,理气中药组方煎剂的效果优于胃复安。     

前房注射卡波姆建立大鼠高眼压模型的观察

目的　前房注射卡波姆建立大鼠高眼压模型，观察卡波姆升眼压效果及对大鼠眼前节和视网膜的影响。方法　随机选取30只SD大鼠，注射前3 d早晚测量基线眼压。右眼定为实验眼，左眼定为对照眼，右眼放出房水后将30 μL的5 g·L^-1卡波姆混悬液注入前房，每日早10时、晚22时在大鼠清醒状态下测量眼压。每周进行双眼眼前节照相并对比。4周末处死26只大鼠（另4只持续观察眼压变化至注射后9周）并取双眼眼球行HE染色，观察实验眼与对照眼视网膜形态，对比视网膜厚度及房角形态。结果　注射前，实验眼白天和夜间眼压分别为（11.10±0.90）mmHg（1 kPa=7.5 mmHg）和（11.92±1.07）mmHg，对照眼分别为（11.22±1.07）mmHg和（11.76±1.08）mmHg；实验眼与对照眼相比，白天、夜间眼压差异均无统计学意义（均为 P＞0.05）；白天与夜间眼压相比，实验眼、对照眼差异均有统计学意义（均为P<0.05）。卡波姆在前房中呈现出弥散型和沉积型两种存在方式，弥散型和沉积型大鼠1周内眼压分别为（17.83±3.54）mmHg和（13.00±1.55）mmHg，两者相比差异具有统计学意义（P＜0.05）。注射后第1天至第19天，实验眼与对照眼白天眼压相比差异均具有统计学意义（均为P＜0.05）；注射后第1天至第27天，实验眼与对照眼夜间眼压相比差异均具有统计学意义（均为P＜0.05）。实验眼视网膜形态发生改变，注射后4周视网膜厚度为（254.70±21.80）μm，与对照眼的（346.73±24.63）μm相比，差异有统计学意义（P=0.00）。实验眼前房充满卡波姆及虹膜的混合成分，紧贴角膜内皮并延伸至房角，堵塞小梁网结构，正常虹膜形态消失；对照眼房角形态正常。结论　前房注射卡波姆建立大鼠高眼压模型，可维持高眼压4周以上，昼夜眼压差异较为明显，夜间眼压较白天更高，4周后视网膜出现高眼压损伤后的表现。     

Simultaneously recorded intracranial and scalp high frequency oscillations help identify patients with poor postsurgical seizure outcome

ObjectiveHigh frequency oscillations (HFO) between 80–500 Hz are markers of epileptic areas in intracranial and maybe also scalp EEG. We investigate simultaneous recordings of scalp and intracranial EEG and hypothesize that scalp HFOs provide important additional clinical information in the presurgical setting.MethodsSpikes and HFOs were visually identified in all intracranial scalp EEG channels. Analysis of correlation of event location between intracranial and scalp EEG as well as relationship between events and the SOZ and zone of surgical removal was performed.Results24 patients could be included, 23 showed spikes and 19 HFOs on scalp recordings. In 15/19 patients highest scalp HFO rate was located over the implantation side, with 13 patients having the highest scalp and intracranial HFO rate over the same region. 17 patients underwent surgery, 7 became seizure free. Patients with poor post-operative outcome showed significantly more regions with HFO than those with seizure free outcome.ConclusionsScalp HFOs are mostly located over the SOZ. Widespread scalp HFOs are indicative of a larger epileptic network and associated with poor postsurgical outcome.SignificanceAnalysis of scalp HFO add clinically important information about the extent of epileptic areas during presurgical simultaneous scalp and intracranial EEG recordings.     

Detection of recombinant and endogenous mouse melatonin receptors by monoclonal antibodies targeting the C‐terminal domain

Melatonin receptors play important roles in the regulation of circadian and seasonal rhythms, sleep, retinal functions, the immune system, depression, and type 2 diabetes development. Melatonin receptors are approved drug targets for insomnia, non‐24‐hour sleep‐wake disorders, and major depressive disorders. In mammals, two melatonin receptors (MTRs) exist, MT₁ and MT₂, belonging to the G protein‐coupled receptor (GPCR) superfamily. Similar to most other GPCRs, reliable antibodies recognizing melatonin receptors proved to be difficult to obtain. Here, we describe the development of the first monoclonal antibodies (mABs) for mouse MT₁ and MT₂. Purified antibodies were extensively characterized for specific reactivity with mouse, rat, and human MT₁ and MT₂ by Western blot, immunoprecipitation, immunofluorescence, and proximity ligation assay. Several mABs were specific for either mouse MT₁ or MT₂. None of the mABs cross‐reacted with rat MTRs, and some were able to react with human MTRs. The specificity of the selected mABs was validated by immunofluorescence microscopy in three established locations (retina, suprachiasmatic nuclei, pituitary gland) for MTR expression in mice using MTR‐KO mice as control. MT₂ expression was not detected in mouse insulinoma MIN6 cells or pancreatic beta‐cells. Collectively, we report the first monoclonal antibodies recognizing recombinant and native mouse melatonin receptors that will be valuable tools for future studies.     

NUDT15 is a key genetic factor for prediction of hematotoxicity in pediatric patients who received a standard low dosage regimen of 6-mercaptopurine

6-Mercaptopurine (6-MP) is commonly used for treatment of acute lymphoblastic leukemia (ALL). The incidence of hematotoxicity caused by this drug is quite high in Asians even using a standard low dosage regimen. The present study was aimed to elucidate the impact of thiopurine S-methyltransferase (TPMT), a nucleoside diphosphate-linked moiety X-type motif 15 (NUDT15), inosine triphosphatase (ITPA) and ATP Binding Cassette Subfamily C Member 4 (ABCC4) polymorphisms on hematotoxicity in pediatric patients who received a standard low starting dose of 6-MP. One hundred and sixty-nine pediatric patients were enrolled and their genotypes were determined. Patients who carried NUDT15¹3 and NUDT15¹2 genotypes were at a 10–15 fold higher risk of severe neutropenia than those of the wild-type during the early months of the maintenance phase. Risk of neutropenia was not significantly increased in patients with other NUDT15 variants as well as in patients with TPMT, ITPA or ABCC4 variants. These results suggest that NUDT15 polymorphisms particularly, NUDT15¹3 and NUDT15¹2, play major roles in 6-MP-induced severe hematotoxicity even when using a standard low dosage of 6-MP and genotyping of these variants is necessary in order to obtain precise tolerance doses and avoid severe hematotoxicity in pediatric patients.     

The oncogenic impact of RNF2 on cell proliferation,invasion and migration through EMT on mammary carcinoma

Mammary carcinoma (MC) is one of most common malignancy in women, and ring finger protein 2 (RNF2) possesses various roles in vast human tumors. In MC tissues as well as in cell lines RNF2 exhibited high expression, had significant association with tumor size, lymph node status, TNM stage, patients’ poor survival, and promoted cell proliferation, colony formation, cell migration and invasion of MC cell lines which was mediated by downregulation of E-cadherin protein. These data reveal that RNF2 protein plays a vital role in the development of MC and may be a potential therapy target of MC.     

Inter-ictal assay of peripheral circulating inflammatory mediators in migraine patients under adjunctive cervical non-invasive vagus nerve stimulation (nVNS): A proof-of-concept study

Objective

To assay peripheral inter-ictal cytokine serum levels and possible relations with non-invasive vagus nerve stimulation (nVNS) responsiveness in migraineurs.

Botulinum toxin blocks mast cells and prevents rosacea like inflammation

Background

Rosacea is a chronic inflammatory skin condition whose etiology has been linked to mast cells and the antimicrobial peptide cathelicidin LL-37. Individuals with refractory disease have demonstrated clinical benefit with periodic injections of onabotulinum toxin, but the mechanism of action is unknown.

A DNA vaccine expressing an optimized secreted FAPα induces enhanced anti-tumor activity by altering the tumor microenvironment in a murine model of breast cancer

Cancer-associated fibroblasts (CAFs), major components of the tumor microenvironment (TME), promote tumor growth and metastasis and inhibit the anti-tumor immune response. We previously constructed a DNA vaccine expressing human FAPα, which is highly expressed by CAFs, to target these cells in the TME, and observed limited anti-tumor effects in the 4T1 breast cancer model. When the treatment time was delayed until tumor nodes formed, the anti-tumor effect of the vaccine completely disappeared. In this study, to improve the safety and efficacy, we constructed a new FAPα-targeted vaccine containing only the extracellular domain of human FAPα with a tissue plasminogen activator signal sequence for enhanced antigen secretion and immunogenicity. The number of CAFs was more effectively reduced by CD8⁺ T cells induced by the new vaccine. This resulted in decreases in CCL2 and CXCL12 expression, leading to a significant decrease in the ratio of myeloid-derived suppressor cells in the TME. Moreover, when mice were treated after the establishment of tumors, the vaccine could still delay tumor growth. To facilitate the future application of the vaccine in clinical trials, we further optimized the gene codons and reduced the homology between the vaccine and the original sequence, which may be convenient for evaluating the vaccine distribution in the human body. These results indicated that the new FAPα-targeted vaccine expressing an optimized secreted human FAPα induced enhanced anti-tumor activity by reducing the number of FAPα⁺ CAFs and enhancing the recruitment of effector T cells in the 4T1 tumor model mice.