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Aims: Hyperhomocysteinemia (HHcy) has been considered as a risk factor for cardiovascular disease (CVD), Alzheimer's disease (AD), nonalcoholic fatty liver (NAFL) and many other pathological condit...

Adiponectin is an adipocyte‐derived hormone, which is closely associated with the development of Alzheimer's disease (AD) and has potential preventive and therapeutic significance. In the present study, we explored the relationship between adiponectin and circadian rhythm disorder in AD, the effect of adiponectin on the abnormal expression of Bmal1 mRNA/protein induced by amyloid‐β protein 31‐35 (Aβ31‐35), and the underlying mechanism of action. We found that adiponectin‐knockout mice exhibited amyloid‐β deposition, circadian rhythm disorders and abnormal expression of Bmal1. Adiponectin ameliorated the abnormal expression of the Bmal1 mRNA/protein caused by Aβ31‐35 by inhibiting the activity of glycogen synthase kinase 3β (GSK3β). These results suggest that adiponectin deficiency could induce circadian rhythm disorders and abnormal expression of the Bmal1 mRNA/protein, whilst exogenous administration of adiponectin may improve Aβ31‐35‐induced abnormal expression of Bmal1 by inhibiting the activity of GSK3β, thus providing a novel idea for the treatment of AD.

Large conductance calcium-activated potassium channel (BK channel) is widely expressed in skeletal muscle, myocardium, smooth muscle and other muscle tissues. Mutation, abnormal expression and altered activity of BK channel are linked to muscle-related diseases such as dyskinesia, epilepsy and erectile dysfunction. In order to compare the effects of BK channel on different muscle tissues, we constructed BK channel gene knockout rats‌‌‌‌‍‍‍ (BK-/- rats). HE staining, open field and grip strength tests, ultrasound, blood pressure measurement and vascular tension test were utilized to explore the effects of BK channel deletion on the structure and function changes in skeletal muscle, myocardium, and vascular smooth muscle (VSM). It was found that compared with wild-type rats, the BK-/- rats showed decreased skeletal muscle fiber area, grip, movement distance and speed at 2 and 12 months of ages. At heart, the muscle fiber area, cardiac systolic/diastolic function and heart rate decreased in BK-/- rats. The wall of the left ventricle became thin. However, the vascular wall of BK-/- rats thickened, the pulse wave velocity was increased, and the VSM contraction was enhanced. Unexpectedly, both systolic and diastolic blood pressure were reduced in BK-/- rats, while pulse pressure difference was increased. These results suggest that BK channel may have different effects on different types of muscle tissue, and it should be noted that different parts of muscle tissue may have different effects when BK channel-related drugs are used.

To observe the therapeutic effect of mild moxibustion on irritable bowel syndrome (IBS) visceral hyperalgesia model rats and its regulatory effect on P2X3 receptors in the spinal cord, anterior cingutate cortex (ACC) and thalamic ventral posterolateral nucleus (VPL). Thirty 8-day-old newborn rats were randomly divided into a normal group (n=6) and a modeling group (n=24) according to the completely random number table method. Rats in the normal group were bred routinely, and those in the modeling group were subjected to preparing IBS chronic visceral hyperalgesia model using colorectal distention (CRD) in stimulation method. Rats successfully modelled were re-divided into a model group, a mild moxibustion group, a P2X3 receptor antagonist group, and a normal saline group according to the completely random number table method with 6 rats in each group. Rats in each group received corresponding interventions from the 37-day old, once a day for 7 consecutive days. Immunohistochemistry and Western blot assays were used to detect P2X3 protein expressions in the spinal cord, ACC and VPL of rats. Under different intensities of CRD stimulation, the abdominal withdrawal reflex (AWR) scores of the model group were significantly increased versus the normal group (all P

Cardiac dysfunction is involved in disorders of energy metabolism. High-titre autoantibodies against the β1 -adrenoceptor (β1 -AAs) have been reported to exist in patients with cardiac dysfunction; however, the mechanism by which β1 -AAs affect cardiac function is unknown. This study aimed to determine whether β1 -AAs disturb myocardium energy metabolism and cause cardiac dysfunction. β1 -AA monoclonal antibodies (β1 -AAmAbs) were successfully pre-synthesized by hybridoma clones and used in all experiments. β1 -AAmAbs impaired cardiac function and induced a myocardial metabolic disturbance, as evidenced by decreased left ventricular ejection fraction and fractional shortening. In addition, β1 -AAmAbs decreased the adenosine triphosphate level and increased cardiac energy consumption (rate-pressure product). We further showed that the effects of β1 -AAmAbs on heart tissue might involve the mitochondria and metabolic pathways via the β1 -adrenoceptor based on an immunoprecipitation and mass spectrometry. Additionally, we found that β1 -AAmAbs impaired myocardial mitochondrial structure, decreased the membrane potential, and induced insufficient mitophagy. In conclusion, β1 -AAmAb-induced cardiac dysfunction is partly due to a disturbance in myocardial energy metabolism.

Sp1-CSE-H2S pathway plays an important role in homocysteine-metabolism, whose disorder can result in hyperhomocysteinemia. H2S deficiency in hyperhomocysteinemia has been reported, while the underlying mechanism and whether it in turn affects the progress of hyperhomocysteinemia are unclear. This study focused on the post-translational modification of Sp1/CSE and revealed four major findings: (1) Homocysteine-accumulation augmented CSE's nitration, inhibited its bio-activity, thus caused H2S deficiency. (2) H2S deficiency inhibited the S-sulfhydration of Sp1, down-regulated CSE and decreased H2S further, which in turn weakened CSE's own S-sulfhydration. (3) CSE was S-sulfhydrated at Cys84, Cys109, Cys172, Cys229, Cys252, Cys307 and Cys310, among which the S-sulfhydration of Cys172 and Cys310 didn't affect its enzymatic activity, while the S-sulfhydration of Cys84, Cys109, Cys229, Cys252 and Cys307 was necessary for its bio-activity. (4) H2S deficiency trapped homocysteine-metabolism into a vicious cycle, which could be broken by either blocking nitration or restoring S-sulfhydration. This study detected a new mechanism that caused severe hyperhomocysteinemia, thereby provided new therapeutic strategies for hyperhomocysteinemia.

Exposure to adverse factors in utero may lead to adaptive changes in cardiac structure and metabolism, which increases the risk of chronic cardiovascular disease later in life. Studies showed that the angiotensin II type 1 receptor autoantibodies (AT1-AAs) are able to cross the placenta into the circulation of pregnant rodents' embryo, which adversely affects embryogenesis. However, the effects of AT1-AA exposure on the fetal heart in utero are still unknown. In this study, we investigated whether intrauterine AT1-AA exposure has adverse effects on fetal heart structure, function and metabolism. AT1-AA-positive pregnant mouse models were successfully established by passive immunity, evidenced by increased AT1-AA content. Morphological and ultrasonic results showed that the fetal mice on embryonic day 18 (E18) of AT1-AA group have loose and disordered myocardial structure, and decreased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), compared with control groups. The myocardium of AT1-AA group fetal mice on E18 exhibited increased expression of the key molecules in the glycolytic pathway, pyruvate and lactic acid content and ATP production, suggesting that the glycolysis rate was enhanced. Furthermore, the enhanced effect of glycolysis caused by AT1-AA is mainly through the PPARβ/δ pathway. These data confirmed that fetus exposure to AT1-AA in utero developed left ventricular dysfunction, myocardial structural arrangement disorders, and enhanced glycolysis on E18. Our results support AT1-AA being a potentially harmful factor for cardiovascular disease in fetal mice.

Background. Electroacupuncture (EA) has been confirmed effectiveness in the treatment of irritable bowel syndrome (IBS), and P2X3 receptors in the peripheral and central neurons participate in the acupuncture-mediated relief of the visceral pain in IBS. Objective. To reveal the neurobiological mechanism that P2X3 receptor of colonic primary sensory neurons in the dorsal root ganglia of the lumbosacral segment is involved in the alleviation of visceral hypersensitivity by EA in an IBS rat model. Methods. The IBS chronic visceral pain rat model was established according to the method of Al-Chaer et al. EA at the bilateral He-Mu points, including ST25 and ST37, was conducted for intervention. The behavioral studies, histopathology of colon, electrophysiology, immunofluorescence histochemistry, and real-time polymerase chain reaction assays were used to observe the role of P2X3 receptor in the colon and related DRG in relieving visceral hypersensitivity by EA. Results. EA significantly reduced the behavior scores of the IBS rats under different levels (20, 40, 60, 80 mmHg) of colorectal distention stimulation and downregulated the expression levels of P2X3 receptor protein and mRNA in colon and related DRG of the IBS rats. EA also regulated the electrical properties of the membranes, including the resting membrane potential, rheobase, and action potential of colon-associated DRG neurons in the IBS rats. Conclusion. EA can regulate the P2X3 receptor protein and mRNA expression levels in the colon and related DRG of IBS rats with visceral pain and then regulate the excitatory properties of DRG neurons.

Background: Recent studies have shown that the pathogenesis of ulcerative colitis (UC) is closely related to the gut microbiota. Moxibustion, a common treatment in traditional Chinese medicine, is the burning of the herb moxa over acupuncture points. Moxibustion has been used to improve the inflammation and gastrointestinal dysfunctions in gastrointestinal disorders such as UC. In this study, we investigated whether moxibustion could improve the gut microbial dysbiosis induced by dextran sulphate sodium (DSS).Methods: Twenty-five male rats were randomly assigned into five groups: normal (NG), UC model (UC), moxibustion (UC+MOX), mesalazine (UC+MES), and normal rats with moxibustion (NG+MOX). The UC rat model was established by administering DSS solution. The rats in the UC+MOX and NG+MOX groups were treated with moxibustion at Tianshu (bilateral, ST25) points once daily for 7 consecutive days, and the UC+MES group rats were treated with mesalazine once daily for 7 consecutive days. After intervention, gut microbiota profiling was conducted by metagenomic high throughput sequencing technology. The gut microbiota composition, diversity and function were analyzed and compared using metagenomics methodologies.Results: The most abundant phyla of five groups were Bacteroidetes, Firmicutes, Proteobacteria and Actinobacteria. Moxibustion treatment increased abundance levels of Bacteroidetes, Actinobacteria, Ascomycota, Synergistetes and decreased abundance of Firmicutes, Proteobacteria. At the genus level, the abundance of Bacteroides, Bacteroides_bacterium_M7, Prevotella, Bacteroidales_bacterium_H2, were increased and Bacteroides_bacterium_H3, Parabacteroides, Porphyromonas, Alistipes, Parasutterella were decreased in the UC group in comparsion with those in the NG group. Moxibustion increased the abundance of Bacteroides and Bacteroides_bacterium_H3 and decreased Bacteroides_bacterium_M7, Prevotella, Bacteroidales_bacterium_H2. In addition, compare with the NG group, genes involved in certain metabolic pathways, such as energy production and conversion, amino acid transport and metabolism, nucleotide transport and metabolism, carbohydrate transport and metabolism, replication, recombination and repair, were under-represented in the UC group, and these changes in the metabolic pathways could be reversed by moxibustion treatment and mesalazine treatment.Conclusion: Our findings suggest that moxibustion treatment may protect the host from mucosal inflammation by modulating the intestinal microbiota community.

Moxibustion therapy is a traditional Chinese medicine external treatment method, which involves crushing dried herb Artemisia argyi H. Lév.Vanio and rolling it into a long cigarette-like strip, igniting it and using its warmth to stimulate specific acupuncture points for a certain period of time. It is often used in Asia to treat various diseases, especially abdominal pain. Clinical reports suggest that acupuncture and moxibustion are the effective treatment for Irritable Bowel Syndrome with Diarrhea (IBS-D). However, there is no placebo-controlled study to prove its safety and efficacy.To evaluate the effects of mild moxibustion (MM) for the treatment of irritable bowel syndrome with diarrhea (IBS-D) through comparisons with those of placebo moxibustion.This was a single-site, randomized controlled trial was conducted at Shanghai Research Institute of Acupuncture and Meridian in China and enrolled 76 participants who met the Rome IV diagnostic criteria for IBS-D between May 2017 and December 2019. 76 participants were randomized to either mild moxibustion (MM) or placebo moxibustion group (PM) in a 1:1 ratio. 18 sessions of MM or PM were implemented over the course of 6 weeks (3 times per week). The primary outcome was adequate relief after 6 weeks of treatment.Of 76 patients with IBS-D who were randomized (38 in the MM group and 38 in the PM group) were included in the intention-to-treat (ITT) analysis set. After treatment at week 6, the response rate was significantly higher in the MM group than the PM group (81.58% vs. 36.84%) with an estimated difference of 44.74 (95% CI, 23.46 to 66.02, P 0.001). No participant reported severe adverse effects.The findings suggest that mild moxibustion may be more effective than placebo moxibustion for the treatment of IBS-D, with effects lasting up to 12 weeks.ChiCTR, ChiCTR2100046852. Registered 29 May 2021 - Retrospectively registered, URL: http://www.chictr.org.cn/showproj.aspx?proj=127000.

During the 2019 novel coronavirus (SARS-CoV-2) outbreak in China (from January 24 to March 11, 2020), our center performed 16 organ transplants (10 kidney, 4 liver, and 2 lung transplants) harvested from deceased donors. Regarding the strategies to prevent infections of SARS-CoV-2, we implemented specific measures for the donor and recipient management, as well as prevention of hospital-acquired infections. All 16 organ recipients had a favorable outcome without SARS-CoV-2 infection. Our approaches aiming to interrupt the spread of SARS-CoV-2 within the transplantation wards were successful, and allowed us to maintain the transplantation program for deceased liver, kidney, and lung organ recipients.

Angiotensin II type 1 receptor autoantibodies (AT1-AA) cause endothelial-dependent smooth muscle relaxation disorder. It is well understood that impairment of the NO–cGMP signaling pathway is one of the mechanisms of endothelial-dependent smooth muscle relaxation disorder. However, it is still unclear whether AT1-AA induces endothelial-dependent smooth muscle relaxation disorder via the impairment of the NO–cGMP signaling pathway. In addition, adiponectin exerts vascular endothelial protection through the NO–cGMP signaling pathway. Therefore, the purpose of this investigation was to assess the mechanism of vascular endothelial-dependent smooth muscle relaxation disorder induced by AT1-AA and the role of adiponectin in attenuating this dysregulation. Serum endothelin-1 (ET-1), adiponectin and AT1-AA were detected by enzyme-linked immunosorbent assay. In preeclamptic patients, there was an increased level of AT1-AA, which was positively correlated with ET-1 and negatively correlated with adiponectin, as elevated levels of ET-1 suggested endothelial injury. AT1-AA-positive animal models were actively immunized with the second extracellular loop of the angiotensin II type 1 receptor (AT1R-ECII) for eight weeks. In thoracic aortas of AT1-AA positive rats, ET-1 was elevated, endothelium-dependent vasodilation was decreased. Paradoxically, as the upstream element of the NO-cGMP signaling pathway, NO production was not decreased but increased, and the ratio of p-VASP/VASP, an established biochemical endpoint of NO-cGMP signaling pathway, was reduced. Moreover, the levels of nitrotyrosine and gp91phox which indicate the presence of peroxynitrite (ONOO-) and superoxide anion (O2·−) were increased. Pretreatment with the ONOO- scavenger FeTMPyP or O2·−scavenger Tempol normalized vasorelaxation. Key enzymes responsible for NO synthesis were also assessed. iNOS protein expression was increased, but p-eNOS(Ser1177)/eNOS was reduced. Preincubation with the iNOS inhibitor 1400 W or eNOS agonist nebivolol restored vasorelaxation. Further experiments showed levels of p-AMPKα (Thr172)/AMPKα, which controls iNOS expression and eNOS activity, to have been reduced. Furthermore, levels of the upstream component of AMPK, adiponectin, was reduced in sera of AT1-AA positive rats and supplementation of adiponectin significantly decreased ET-1 contents, improved endothelial-dependent vasodilation, reduced NO production, elevated p-VASP/VASP, inhibited protein expression of nitrotyrosine and gp91phox, reduced iNOS overexpression, and increased eNOS phosphorylation at Ser1177 in the thoracic aorta of AT1-AA positive rats. These results established that impairment NO–cGMP pathway may aggravate the endothelial-dependent smooth muscle relaxation disorder in AT1-AA positive rats and adiponectin improved endothelial-dependent smooth muscle relaxation disorder by enhancing NO–cGMP pathway. This discovery may shed a novel light on clinical treatment of vascular diseases associated with AT1-AA.

Numerous studies have reported significantly elevated titers of serum autoantibody against the second extracellular loop of β1-adrenoceptor (β1-AA), a catecholamine-like substance with β1-adrenergic activity, in patients with heart failure. Although evidence demonstrates that this autoantibody may alter T cell proliferation and secretion, the role of T lymphocytes in heart failure induced by β1-AA remains unclear. The current study was designed to determine whether T cell disorder contributes to heart failure induced by β1-AA. β1-AA monoclonal antibodies (β1-AAmAb) produced using the hybridoma technique were administered in wild-type mice or T lymphocyte deficiency nudes for 12 weeks. T lymphocytes from heart failure patients and neonatal cardiomyocytes were utilized in vitro. Mouse protein antibody array analysis was employed to detect the cytokines responsible for β1-AAmAb-induced heart failure. Compared to wild-type mice, T lymphocyte deficiency mice prevented cardiac function from getting worse, attenuated adverse remodeling, and ameliorated cardiomyocyte apoptosis and fibrosis. As shown by protein array, the serum level of interleukin (IL)-6 was significantly lower in the nude group as compared to wild-type after β1-AAmAb treatment. Mechanistic studies in vitro demonstrated that T lymphocyte culture supernatants stimulated by β1-AAmAb caused direct damage in the cardiomyocytes, and β1-AAmAb promoted proliferation of T lymphocytes isolated from patients with heart failure and increased IL-6 release. IL-6-specific siRNA virtually abolished cardiomyocyte apoptosis, suggesting that IL-6 may be a key cytokine released by T lymphocytes and responsible for β1-AAmAb-induced cardiac remodeling. Collectively, we demonstrate that β1-AAmAb-induced cardiac remodeling via mediating T lymphocyte disorder and releasing a variety of IL-6.

Autoantibody against β1-adrenoceptor (β1-AA) has been shown to be closely linked to the aggravation of heart failure. Removal of β1-AA remarkably attenuated patients' cardiac dysfunction. We found that β1-AA induced rat heart failure with increased CD4+ T cells. However, whether or not β1-AA interacts with T cells isolated from heart failure patients remains unknown. Twenty-one β1-AA-negative heart failure patients were divided into those taking β-adrenergic blocker and those not. The effects of β1-AA monoclonal antibodies (β1-AAmAb) on T cells proliferation were detected using the CCK-8 assay. IFN-γ and IL-4 production by human T cells were measured by after the administration of β1-AAmAb. The levels of cardiomyocyte apoptosis and hypertrophy were detected after co-cultured with the supernatant of T cells pre-stimulated by β1-AAmAb. It was found that β1-AAmAb promoted T cell proliferation via the β1-AR/cAMP/PKA pathway in patients who not take β-blocker. β1-AAmAb inhibited the characteristic cytokine secretion of Th1, IFN-γ, but had no significant effect on the Th2 cytokine IL-4. Supernatant resulted from the T cells pre-treated with β1-AAmAb induced cardiomyocytes remodeling, as evidenced by increased levels of cardiomyocytes apoptosis and hypertrophy. We propose that heart failure is likely to be an interference factor for Th-mediated immunity, and the presence of β1-AAmAb may aggravate this effect and deteriorate concomitant inflammatory injury in cardiomyocytes, partially via β1-AR/cAMP/PKA pathway.

Background and Aims. Intestinal fibrosis is one of the severe and common complications of Crohn’s disease (CD), but the etiology and pathogenesis remain uncertain. The study intended to examine whether the effect of herb-partitioned moxibustion on rats with CD-associated intestinal fibrosis is associated with the RhoA/ROCK1/MLC pathway. Methods. All experimental rats were randomly allocated into the normal control group (NC), model control group (MC), and herb-partitioned moxibustion group (HPM). Intestinal fibrosis was established in rats with CD by repeated rectal administrations of 2,4,6-trinitrobenzenesulfonic acid (TNBS). Herb-partitioned moxibustion was applied at the Qihai (CV6) and Tianshu (ST25) acupoints once daily for 10 days in the HPM group. In this study, histological changes were examined by hematoxylin and eosin (HE) staining; then, Masson’s trichrome staining was used to assess the degree of fibrosis in each group. Experimental methods of immunohistochemistry, western blotting, and real-time PCR were applied to detect the levels of α-SMA, collagen III, RhoA, ROCK1, and p-MLC. Moreover, the double immunofluorescent staining for the colocalization of both α-SMA and ROCK1 was performed. Results. Contrasted with the normal controls, the collagen deposition and fibrosis scores were increased in colonic tissue of model rats, and HPM decreased the collagen deposition and fibrosis scores. The protein of α-SMA and collagen III in the MC group exceeds that of the NC group; HPM decreased the expression of α-SMA and collagen III in rats with intestinal fibrosis. Similarly, the expression of RhoA, ROCK1, and p-MLC in model rats was obviously increased compared with normal controls; the expression of RhoA, ROCK1, and p-MLC was decreased after HPM. The coexpression of α-SMA and ROCK1 in rats with intestinal fibrosis was higher than normal rats. Conclusion. HPM improves CD-associated intestinal fibrosis by suppressing the RhoA/ROCK1/MLC pathway.

Purpose The cardiac disease is now as the leading cause of morbidity and mortality in DMD patients. The early detection of cardiac involvement is challenging and the frequency and presence of the cardiac involvement in the first decade is uncertain. The aim of the current study was to characterize the frequency, pattern and extent of LGE in the first decade in DMD patients, and identify a set of prognosis factors that portend the presence and extent of LGE in those patients. Methods There are 93 DMD patients (mean age: 8.31±2.30 years, rang: 3–13 years) with a biopsy showing absent dystrophin and/or DNA analysis demonstrating a characteristic dystrophin mutation enrolled. All patients underwent a complete cine imaging and LGE imaging. The left ventricular function and tissue characteristic were analyzed and statistic. Results There were 42% patients presented LGE-positive. Six years old was the youngest age onset myocardial fibrosis, and after 7 years old, the frequency of LGE positive rising fast. Moreover, all LGE positive patients showed complex patterns with non-ischaemic pattern, including septum, papillary muscle and right ventricular involvement. The free wall, inferior, apical and septum were the most common position. The papillary and right ventricular were rarely common. And all patients presented right ventricular involvement associated with septum involvement. Moreover, compared with younger patients ( Conclusion The myocardial fibrosis could onset in DMD patients in the first decade with a rapid rising and presented with complex patterns. The early CMR examination is necessary for the assessment of cardiac involvement and should consider to complete in younger age in DMD patients. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): The National Natural Science Foundation

Background Traditional Chinese medicine (TCM) has a long history of use in breast cancer, but lacking systematic evidence to support its clinical benefits. In this study, we evaluated the prophylactic and therapeutic effects of moxibustion combined with decoctions for treating chemotherapy-induced myelosuppression (CIM) in early-stage breast cancer patients. Methods This is a randomized controlled clinical trial single-blinded for TCM decoction but not moxibustion. Patients are equally divided into the control group without decoction and moxibustion treatment (control), the decoction+moxibustion group (MD), and the placebo+moxibustion group (MP), according to the following stratification factors: age (below 40s, 40s, 50s, and 60s or above), chemotherapy regimen (anthracyclines, taxanes, anthracyclines+taxane, and others), and chemotherapy strategy (adjuvant and neoadjuvant). The TCM decoction is Wenshen Shengbai Decoction. The anticipated sample size is 462 cases (154 cases in each group). All participants are expected to treat with chemotherapy and recombinant human granulocyte colony-stimulating factor (rhG-CSF). The primary outcomes include the proportion of patients with relief of leukopenia and/or neutropenia, the myelosuppression-associated serious adverse event including grade 3–4 leukopenia and/or neutropenia, and febrile neutropenia, and the dose of rhG-CSF. The secondary outcomes include chemotherapy adherence, stratified analysis, adverse reactions, quality of life by EORTC Breast-Cancer-Specific Quality of Life Questionnaire including EORTC QLQ-C30 (V3.0) and QLQ-BR23, TCM Constitution, and 3-year disease-free survival and overall survival. Baseline information including age, surgical approach, chemotherapy regimen and strategy, pathological stage, and molecular subtype will be recorded. Discussion This will be the first randomized controlled trial to evaluate the efficacy of moxibustion combined with TCM decoction in treating CIM in early-stage breast cancer patients, aiming to standardize the TCM decoction and moxibustion method, thus providing evidence for its clinical benefit. Trial registration chictr.org.cn ChiCTR-INR-16009557. Registered on 23 October 2016.

Background Voltage-gated K+ (Kv) channels in coronary artery smooth muscle cells (CSMCs), especially the major specific Kv1 subfamily, contribute to coronary artery vasodilation. Advanced glycation end products (AGEs) have been strongly implicated in diabetes-related cardiovascular complications. Our previous study showed AGEs can impair Kv channel-mediated coronary vasodilation by reducing Kv channel activity. However, its underlying mechanism remains unclear. Purpose Here, we used isolated rat small coronary arteries (RSCAs) and primary CSMCs to investigate the effect of AGEs on Kv channel-mediated coronary vasodilation and the possible involvement of peroxisome proliferators-activated receptor (PPAR)-γ pathway. Methods RSCAs and primary CSMCs were isolated, cultured and treated with bovine serum albumin (BSA), AGE-BSA, alagrebrium (ALA, AGE cross-linking breaker), pioglitazone (PIO) and/or GW9662, and then divided into the following groups: DMEM, BSA, AGE, AGE+ALA, AGE+PIO, and AGE+PIO+GW9662. Kv channel-mediated coronary vasodilation was analyzed using wire myograph. Histology and immunohistochemistry of RSCAs were performed. Western blot was used to detect the protein expression of RAGE, the major Kv1 channel subunits expressed in CSMCs (Kv1.2/1.5), PPAR-γ, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-2 (NOX-2). Results AGEs markedly reduced forskolin-induced Kv channel-mediated vasodilation of RSCAs by interacting with the receptor for AGEs (RAGE), and ALA or PIO significantly reversed this effect. In both RSCAs and primary CSMCs, AGEs decreased Kv1.2 and Kv1.5 channel protein expression, inhibited PPAR-γ expression, increased RAGE and NOX-2 expression. Treatment with ALA or PIO partially reversed the effects of AGEs on Kv1.2/Kv1.5 expression, accompanied by elevation of PPAR-γ level and diminished oxidative stress. Conclusion AGE/RAGE axis-induced inhibition of PPAR-γ pathway and enhancement of oxidative stress may contribute to AGEs-mediated Kv channel dysfunction and coronary vasodilation in RSCAs. Our results may provide new insights into developing therapeutic strategies to manage diabetic vasculature. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): National Natural Science Foundation of China; Natural Science Foundation of Beijing (7172059)

Objective. To explore whether the effect of electroacupuncture (EA) on visceral hypersensitivity (VH) in rats with irritable bowel syndrome (IBS) is related to the changes of astrocyte activation in the medial thalamus (MT) and anterior cingulate cortex (ACC).Method. Male Sprague-Dawley rats were randomly divided into the normal control (NC) group, model control (MC) group, electroacupuncture (EA) group, and fluorocitrate (FCA) group. A model of visceral hypersensitivity was established by neonatal colorectal irritation. In the EA group, needles were inserted into the skin at the Tianshu (ST25) and Shangjuxu (ST37) acupoints, once a day for 7 days. The FCA group received intrathecal injection of FCA on the 1st, 4th, and 7th days. Visceral hypersensitivity was evaluated by the abdominal withdrawal reflex (AWR), and glial fibrillary acidic protein (GFAP) mRNA and protein levels in the MT and ACC were detected by real-time PCR, immunohistochemistry, and western blots.Results. The AWR score in the MC group was significantly higher than in the NC group, and EA and FCA reduced the AWR score of VH rats. GFAP mRNA and protein levels in the MT and ACC of rats in the MC group were significantly increased compared with the NC group. After either electroacupuncture or fluorocitrate, GFAP mRNA and protein levels in the MT and ACC were both clearly reduced.Conclusion. Electroacupuncture alleviates IBS visceral hypersensitivity by inhibiting the activation of astrocytes in the MT and ACC.

Background: The autoimmune mechanism in coronary heart disease (CHD) has been concerned recently. It is reported that the second extracellular loop of angiotensin II type 1 receptor (AT1R-ECII) autoantibody (AT1-AA) presents in the serum of CHD patients, but its clinical significance is unclear. The purpose of this study was to identify the diverse functional subclasses of AT1-AA in the patients of CHD. Methods: The AT1-AA levels in sera of 306 CHD patients were detected by ELISA. Then AT1-AA was purified from each patient’s serum (n=127) and their effects on intracellular calcium ([Ca2+]i) of mouse arterial smooth muscle cells (MASMC) was detected to classify the subclasses of AT1-AA from patients. Correlation analysis and logistic regression were used to analyze the clinical significances of different subclasses of AT1-AA in CHD. To further confirm AT1-AA classification, rats were subcutaneously injected with AT1R-ECII peptide. Findings: There were 4 [Ca2+]i change curves indicating different functional subclasses of AT1-AA (H1-, H2-, H3-, and H4-AT1-AA). Correlation analysis showed H1- and H2-AT1-AA levels were positively associated with endogenous and exogenous coagulation, respectively; H3-AT1-AA had no correlation; H4-AT1-AA was negatively correlated with leukocyte number and bile acid. Logistic regression showed H2-AT1-AA had predictive value for severe CHD. Moreover, H1- and H2-AT1-AA exerted cytotoxic effects in vitro, while H4-AT1-AA increased cell viability. 4 similar AT1-AA subclasses were found in AT1-AA-positive rats. Interpretation: This study demonstrates that there are 4 kinds of functional AT1-AA with different clinical significance in CHD patients, suggesting the importance of AT1-AA subclasses distinguishment in cardiovascular disease. Funding Statement: This work is supported by the Beijing Natural Science Foundation Program and Scientific Research Key Program of Beijing Municipal Commission of Education (No. KZ201810025039); the National Natural Science Foundation of China (No. 31771267); the Open Fund from Key Laboratory of Cellular Physiology (Shanxi Medical University), Ministry of Education, China (No. KLMEC/SXMU-201902). Declaration of Interests: The authors have declared that no conflict of interest exists. Ethics Approval Statement: All human studies had been conducted according to Declaration of Helsinki principles and approved by the local Research Ethics Committee (Beijing Anzhen Hospital, Capital Medical University, Beijing, China) (Ethics No.: 2019023X). All patients were identified by number and signed informed consent. All the animal experiments were approved by the Institutional Animal Care and Use Committee and Ethics Committee of Capital Medical University.

Insulin resistance is highly associated with an adverse intrauterine environment. We previously reported that fetal rats exposed to angiotensin II type 1 receptor (AT1R) autoantibody (AT1-AA) displayed increased susceptibility to metabolic diseases during middle age. However, the timing of the onset of insulin resistance remains unknown. In this study, we examined the offspring of AT1-AA-positive rats, tracking the development of insulin resistance. Pregnant rats were intravenously injected with AT1-AA. Afterwards, we collected serum samples and liver tissues of the offspring at various stages, including gestation day 18, 3 weeks (weaning period), 18 weeks (young adulthood), and 48 weeks (middle age) after birth. Compared with saline control group, hepatic vacuolar degeneration was visible in AT1-AA offspring rats as early as 3 weeks; hyperinsulinemia and impaired glucose tolerance occurred at 18 weeks of age, however, insulin resistance was not observed until 48 weeks. At 18 weeks we detected suppressed protein levels of insulin receptor (IR) but increased levels of IR substrate 1 (IRS1) in the liver of AT1-AA group rats. Interestingly, both IR and IRS1/2 were significantly decreased at 48 weeks. Liver proteomic analysis indicated that the differences in protein expression between the AT1-AA and control rats became more pronounced with age, particularly in terms of mitochondrial energy metabolism. Rats exposed to AT1-AA in utero developed hyperinsulinemia from young adulthood which subsequently progressed to insulin resistance, and was linked with abnormal hepatic structure and impaired IR signaling. Additionally, dysregulation of energy metabolism may play a fundamental role in predisposing offspring to insulin resistance.

High titer of β1-adrenoreceptor autoantibodies (β1-AA) has been reported to appear in heart failure patients. It induces sustained β1-adrenergic receptor (β1-AR) activation which leads to heart failure (HF), but the mechanism is as yet unclear. In order to investigate the mechanisms causing β1-AR non-desensitization, we studied the beating frequency of the neonatal rat cardiomyocytes (NRCMs) under different conditions (an injection of isoprenaline (ISO) for one group and β1-AA for the other) and established three dynamic models in order to best describe the true relationships shown in medical experiments; one model used a control group of healthy rats; then in HF rats one focused on conformation changes in β1-AR; the other examined interaction between β1-AR and β2-adrenergic receptors (β2-AR). Comparing the experimental data and corresponding Akaike information criterion (AIC) values, we concluded that the interaction model was the most likely mechanism. We used mathematical methods to explore the mechanism for the development of heart failure and to find potential targets for prevention and treatment. The aim of the paper was to provide a strong theoretical basis for the clinical development of personalized treatment programs. We also carried out sensitivity analysis of the initial concentration β1-AA and found that they had a noticeable effect on the fitting results.

Aims β1-adrenergic receptor autoantibodies (β1-AAs) and β2-adrenergic receptor autoantibodies (β2-AAs) are present in patients with heart failure (HF); however, their interrelationship with cardiac structure and function remains unknown. This study explored the effects of the imbalance between β1-AAs and β2-AAs on cardiac structure and its underlying mechanisms in HF. Methods and results Patients with left systolic HF who suffered from coronary heart disease (65.9%) or dilated cardiomyopathy (34.1%) were divided into New York Heart Association Classes I-II (n = 51) and Classes III-IV (n = 37) and compared with healthy volunteers as controls (n = 41). Total immunoglobulin G from HF patient serum comprising β1-AAs and/or β2-AAs were determined and purified for in vitro studies from neonatal rat cardiomyocytes (NRCMs). In addition, HF was induced by doxorubicin in mice. We observed that the increased ratio of β1-AAs/β2-AAs was associated with worsening HF in patients. Moreover, β2-AAs from patients with HF suppressed the hyper-shrinking and apoptosis of NRCMS induced by β1-AAs from some patients. Finally, β2-AAs alleviated both myocardial damage and β1-AAs production induced by doxorubicin in mice. Conclusion β2-AAs were capable of antagonizing the effects imposed by β1-AAs both in vitro and in vivo. The imbalance of β1-AAs and β2-AAs in patients with HF is a mechanism underlying HF progression, and the increasing ratio of β1-AAs/β2-AAs should be considered a clinical assessment factor for the deterioration of cardiac function in patients with HF.

Preeclamptic women are reported to have inadequate plasma volume expansion coupled with a suppressed secretion of aldosterone; however, the specific mechanism of preeclampsia remains unclear. We demonstrated that the presence of long-term angiotensin II type 1 receptor autoantibody (AT1-AA) reduces aldosterone production by triggering a Ca2+ overload in H295R cells. AT1-AA was discovered in preeclamptic women and reported to activate AT1R, and consequently elevate intracellular Ca2+. We found that AT1-AA significantly prolonged the time of intracellular Ca2+ elevation. Besides promoting aldosterone production as a second messenger, Ca2+ overload shows a cytotoxic effect. Our data reveals that long-term presence of AT1-AA triggered a Ca2+ overload and consequent impairment of aldosterone production, which could be prevented by a PKC inhibitor, Go 6983, or a calcium channel inhibitor, nifedipine. These findings have clinical significance because AT1R blockers are not recommended for treatment of preeclampsia due to their potential harm to the fetus. Our findings also emphasize a potential clinical benefit of immunoadsorption or neutralization of AT1-AA in preeclamptic women.

// Chunhui Bao 1, * , Peng Liu 2, * , Yin Shi 3 , Luyi Wu 1 , Xiaoming Jin 4 , Xiaoqing Zeng 5 , Jianye Zhang 6 , Di Wang 1 , Huirong Liu 3 and Huangan Wu 1 1 Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai, China 2 Life Sciences Research Center, School of Life Sciences and Technology, Xidian University, Xi’an, China 3 Outpatient Department, Shanghai Research Institute of Acupuncture and Meridian, Shanghai University of Traditional Chinese Medicine, Shanghai, China 4 Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, USA 5 Department of Gastroenterology, Zhongshan Hospital, Fudan University, Shanghai, China 6 Department of Radiology, Shanghai Mental Health Center, Shanghai Jiaotong University School of Medicine, Shanghai, China * These authors contributed equally to this work Correspondence to: Huangan Wu, email: wuhuangan@126.com Huirong Liu, email: lhr_tcm@139.com Keywords: magnetic resonance imaging, Crohn’s disease, gray matter, brain, pain Received: June 16, 2017 Accepted: September 08, 2017 Published: September 22, 2017 ABSTRACT Increasing evidence indicates that abnormal pain processing is present in the central nervous system of patients with Crohn’s disease (CD). The purposes of this study were to assess changes in gray matter (GM) volumes in CD patients in remission and to correlate structural changes in the brain with abdominal pain. We used a 3.0 T magnetic resonance scanner to examine the GM structures in 21 CD patients with abdominal pain, 26 CD patients without abdominal pain, and 30 healthy control subjects (HCs). Voxel-based morphometric analyses were used to assess the brain GM volumes. Patients with abdominal pain exhibited higher CD activity index and lower inflammatory bowel disease questionnaire scores than those of the patients without abdominal pain. Compare to HCs and to patients without abdominal pain, patients with abdominal pain exhibited lower GM volumes in the insula and anterior cingulate cortex (ACC); whereas compare to HCs and to patients with abdominal pain, the patients without abdominal pain exhibited higher GM volumes in the hippocampal and parahippocampal cortex. The GM volumes in the insula and ACC were significantly negatively correlated with daily pain scores. These results suggest that differences exist in the brain GM volume between CD patients in remission with and without abdominal pain. The negative correlation between the GM volumes in the insula and ACC and the presence and severity of abdominal pain in CD suggests these structures are closely related to visceral pain processing.

To explore the beneficial regulatory effect of mild moxibustion from different distances at Zusanli (ST 36) of healthy population on the functions of temperature-related brain regions. In 20 recruited healthy subjects, the change of the temperature-related brain regions induced by mild moxibustion from different distances at Zusanli (ST 36) was observed by functional magnetic resonance imaging (fMRI). In comparison of the values in amplitude of low-frequency fluctuation (fALFF) during and before moxibustion, it has been found that in moxibustion of 2 cm distance, fALFF value increased in the brain regions of the left anterior cingulated cortex and lateral surrounding cerebral regions, and fALFF value decreased in the cerebral regions of the peripheral cortex of the calcarine fissure; in moxibustion of 3 cm distance, fALFF value increased in the brain regions of the right and medial side and paracingulated gyrus, and fALFF value decreased in the cerebral zone of the left middle temporal gyrus; in moxibustion of 4 cm distance, fALFF value increased in the brain regions of the right and medial and paracingulated gyrus; and in moxibustion of 5 cm distance, fALFF value increased in the brain regions of the left hippocampus. In comparison of the value of regional homogeneity (ReHo), it has been found that in moxibustion of 2 cm distance, ReHo value increased in the cerebral zone of the posterior lobe of the right cerebellum, and ReHo value decreased in the cerebral zone of the right occipital lobe; in moxibustion of 3 cm distance, ReHo value increased in the brain regions of the left cerebellar posterior lobe and left frontal lobe, and ReHo value decreased in the cerebral zone of the right inferior temporal gyrus; in moxibustion of 4 cm distance, ReHo value increased in the brain regions of the right superior frontal gyrus and ReHo value decreased in the brain regions of the right parietal lobe and angular gyrus; in moxibustion of 5 cm distance, ReHo value increased in the cerebral zone of the right frontal lobe and ReHo value decreased in the cerebral zone of the right brainstem. In moxibustion of 3 cm distance, the changes in the brain regions basically conform to the transmission route of body trunk temperature.

To explore the correlation between moxibustion sensation and distance of moxa stick and provide reference for clinical practice. A total of 16 healthy volunteers aged 18-35 years old in college were recruited and given mild moxibustion at Shousanli (LI 10), Zusanli (ST 36), Shenshu (BL 23) and Tianshu (ST 25) with moxa stick, and the occurrence and frequency of moxibustion sensation were recorded at distances of 5 cm, 4 cm, 3 cm and 2 cm. Mild moxibustion scale was used to count the score. Warm was the main moxibustion sensation, burning pain and soreness decreased with the rise of distance; for the same acupoint, score of mild moxibustion scale increased with the decrease of distance; score ranged between 5.5 and 6.5 at distance 3 cm, which was the most comfortable distance for volunteers. The distance of 3 cm is the most comfortable distance in mild moxibustion

Although the contributions of sitagliptin to endothelial function in diabetes mellitus were previously reported, the potential mechanisms still remain undefined. Our research was intended to explore the underlying mechanisms of protective effects of sitagliptin treatment on endothelial dysfunction in Zucker diabetic fatty (ZDF) rats. Male lean nondiabetic Zucker rats were used as control and male obese ZDF rats were randomly divided into ZDF and ZDF + sitagliptin groups. The significant decrease in endothelium-dependent relaxation induced by acetylcholine was observed in mesenteric arteries and thoracic aorta rings of ZDF rats. The administration of sitagliptin restored the vascular function effectively. The morphology study showed severe endothelial injuries in thoracic aortas of ZDF rats, and sitagliptin treatment attenuated these changes. The increased malondialdehyde levels and decreased superoxide dismutase activities in serum of ZDF rats were reversed by sitagliptin treatment. Sitagliptin also increased the expression of endothelial nitric oxide synthase and microtubule-associated protein 1 light chain 3 (LC3) and decreased the expression of inducible nitric oxide synthase, 3-nitrotyrosine, and p62 in ZDF rats. After giving Fe (III) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride porphyrin pentachloride (FeTMPyP, a peroxynitrite [ONOO−] scavenger) or sitagliptin to high-glucose (30 mmol/L, 48 hours) cultured human umbilical vein endothelial cells (HUVECs), the increased levels of Beclin-1 and lysosome-associated membrane protein type 2 were detected. Both FeTMPyP and sitagliptin also significantly increased the number of mRFP-GFP-LC3 dots per cell, suggesting that autophagic flux was increased in HUVECs. Our study indicated that sitagliptin treatment can improve the endothelium-dependent relaxation and attenuate the endothelial impairment of ZDF rats. The protective effects of sitagliptin are possibly related to antiperoxynitrite and promoting autophagy.

Sp1 (Specificity protein 1)-CSE (cystathionine-γ-lyase)-H2S (hydrogen sulfide) pathway plays an important role in homocysteine-metabolism, whose disorder can result in hyperhomocysteinemia. The deficiency of plasma H2S in patients and animal models with hyperhomocysteinemia has been reported but it is unclear whether this deficiency plays a role in the progress of hyperhomocysteinemia. Furthermore, it remains unknown whether the post-translational modification of Sp1 or CSE mediated by hyperhomocysteinemia itself can in turn affect the development of hyperhomocysteinemia. By both in vivo and in vitro studies, we conducted immunoprecipitation and maleimide assays to detect the post-translational modification of Sp1-CSE-H2S pathway and revealed four major findings: (1) the accumulation of homocysteine augmented the nitration of CSE, thus blunted its bio-activity and caused H2S deficiency. (2) H2S deficiency lowered the S-sulfhydration of Sp1 and inhibited its transcriptional activity, resulted in lower expression of CSE. CSE deficiency decreased the H2S level further, which in turn lowered the S-sulfhydration level of CSE. (3) CSE was S-sulfhydrated at Cys84, Cys109, Cys172, Cys229, Cys252, Cys307 and Cys310 under physiological conditions, mutation of Cys84, Cys109, Cys229, Cys252 and Cys307 decreased its S-sulfhydration level and bio-activity. (4) H2S deficiency could trap hyperhomocysteinemia into a progressive vicious circle and trigger a rapid increase of homocysteine, while blocking nitration or restoring S-sulfhydration could break this circle. In conclusion, this study reveals a novel mechanism involved in the disorder of homocysteine-metabolism, which may provide a candidate therapeutic strategy for hyperhomocysteinemia.

Background Fatigue is one of the primary symptoms in lung cancer, with a prevalence of 88.0% in survivors of cancer, and an even higher prevalence post resection surgery. Effective fatigue control after lung cancer surgery is important for patient recovery and quality of life. Some studies have shown that acupuncture might be effective in treating cancer-related fatigue; however, randomized controlled trials (RCTs) of suitable sample size are limited. Method/design This is a multi-center, patient-blinded RCT. A total of 320 eligible patients will be recruited in four centers and randomly assigned to either the acupuncture group or the sham acupuncture group in a 1:1 ratio. Treatment will be given twice per week for 12 sessions. Treatment will be given at acupoints GV20, GV29, CV12, CV6, CV4, and bilateral LI4, LR3, SP6, ST36. The primary outcome will be assessed using the Chinese version of The Brief Fatigue Inventory. The secondary outcomes will be measured using The European Organization for Research and The Treatment of Cancer Quality of Life Questionnaire, and the Hamilton Rating Scale for Depression. The primary outcome will be assessed at all main points (baseline, the 3rd week, the 6th week, and at follow up time points) and the secondary outcomes will be assessed at baseline and the 6th week. Intention-to-treat analysis will be used in this RCT. Discussion This trial protocol provides an example of the clinical application acupuncture treatment in the management of lung cancer-related fatigue. If the acupuncture treatment protocol confirms that acupuncture is an effective and safe option for lung cancer-related fatigue, it can be adopted as a standardized treatment. Trial registration Chinese Clinical Trial Registry, ChiCTR1900022831. Registered on 27 April 2019. URL: http://www.chictr.org.cn/showproj.aspx?proj=37823

Background Angiotensin II type 1 receptor ( AT 1 R) autoantibody ( AT 1‐ AA ) was first identified as a causative factor in preeclampsia. Unlike physiological ligand angiotensin II (Ang II ), AT 1‐ AA can induce vasoconstriction in a sustained manner, causing a series of adverse effects, such as vascular injury and poor placental perfusion. However, its underlying mechanisms remain unclear. Here, from the perspective of AT 1 R internalization, the present study investigated the molecular mechanism of sustained vasoconstriction induced by AT 1 R autoantibody. Methods and Results In the current study, we used the vascular‐ring technique to determine that AT 1‐ AA ‐positive IgG, which was obtained from the sera of preeclamptic patients, induced long‐term vasoconstriction in endothelium‐intact or endothelium‐denuded rat thoracic arteries. The effect was caused by prolonged activation of AT 1 R downstream signals in vascular smooth muscle cells, including Ca 2+ , protein kinase C, and extracellular signal‐regulated kinase 1 and 2. Then, using subcellular protein fractionation, cell surface protein biotinylation, and total internal reflection fluorescence, we found that AT 1‐ AA ‐positive IgG resulted in significantly less AT 1 R internalization than in the Ang II treatment group. Moreover, through use of fluorescent tracing and bioluminescence resonance energy transfer, we found that AT 1‐ AA ‐positive IgG cannot induce the recruitment of β‐arrestin1/2, which mediated receptor internalization. Then, the effect of sustained AT 1 R activation induced by AT 1‐ AA ‐positive IgG was rescued by overexpression of β‐arrestin2. Conclusions These data suggested that limited AT 1 R internalization resulting from the inhibition of β‐arrestin1/2 recruitment played an important role in sustained vasoconstriction induced by AT 1‐ AA ‐positive IgG, which may set the stage for avoiding AT 1 R overactivation in the management of preeclampsia.

Background Autoantibodies against the second extracellular loop of the β 1 ‐adrenoceptor (β 1 ‐ AA ) act similarly to agonist of β 1 ‐adrenergic receptor, which plays an important role in the pathophysiological characteristics of ventricular remodeling. Recently, considerable lines of evidence have suggested that CTRP9 (C1q tumor necrosis factor–related protein 9) is a potent cardioprotective cardiokine and protects the heart from ventricular remodeling. The aim of this study was to determine the role of CTRP 9 in ventricular remodeling induced by β 1 ‐ AA . Methods and Results Blood samples were collected from 131 patients with coronary heart disease and 131 healthy subjects. The serum levels of β 1 ‐ AA and CTRP 9 were detected using ELISA . The results revealed that CTRP 9 levels in β 1 ‐ AA –positive patients were lower than those in β 1 ‐ AA –negative patients, and serum CTRP 9 concentrations were inversely correlated with β 1 ‐ AA . β 1 ‐ AA monoclonal antibodies (β 1 ‐ AA mAbs) were administered in mice with and without rAAV 9‐ cTnT ‐Full Ctrp9‐ FLAG virus for 8 weeks. Reverse transcription–polymerase chain reaction/Western analysis showed that cardiomyocyte CTRP 9 expression was significantly reduced in β 1 ‐ AA mAb–treated mice. Moreover, compared with the β 1 ‐ AA mAb alone group, cardiac‐specific CTRP 9 overexpression improved cardiac function, attenuated adverse remodeling, and ameliorated cardiomyocyte apoptosis and fibrosis. Mechanistic studies demonstrated that CTRP 9 overexpression decreased the levels of G‐protein–coupled receptor kinase 2 and promoted the activation of AMP‐dependent kinase pathway. However, cardiac‐specific overexpression of CTRP 9 had no effect on the levels of cAMP and protein kinase A activity elevated by β 1 ‐AAmAb. Conclusions This study provides the first evidence that the long‐term existence of β 1 ‐ AA mAb suppresses cardiac CTRP 9 expression and exaggerates cardiac remodeling, suggesting that CTRP 9 may be a novel therapeutic target against pathologic remodeling in β 1 ‐ AA –positive patients with coronary heart disease.

NEW FINDINGS What is the central question of this study? High titres of autoantibodies against the second extracellular loop of the β1 -adrenergic receptor (β1 -AAs) can be detected in the sera of patients with ventricular arrhythmias, but a causal relationship between β1 -AAs and ventricular arrhythmias has not been established. What is the main finding and its importance? Monoclonal β1 -AAs (β1 -AR mAbs) were used in the experiments. We showed that β1 -AR mAbs increased susceptibility to ventricular arrhythmias and induced repolarization abnormalities. Antibody adsorption of β1 -AAs will be a potential new therapeutic strategy for ventricular arrhythmias in patients with high titres of β1 -AAs. High titres of autoantibodies against the second extracellular loop of the β1 -adrenergic receptor (β1 -AAs) can be detected in sera from patients with ventricular arrhythmias, but a causal relationship between β1 -AAs and ventricular arrhythmias has not been established. In this work, ECGs of guinea-pigs and isolated guinea-pig hearts were recorded. Ventricular tachycardia (VT) and ventricular fibrillation (VF) were evoked by programmed electrical stimulation of the left ventricular epicardium of isolated guinea-pig hearts. The monophasic action potential and effective refractory period of the left ventricle were recorded in paced isolated guinea-pig hearts. Furthermore, to increase the specificity, monoclonal autoantibodies against the second extracellular loop of the β1 -adrenergic receptor (β1 -AR mAbs) were used in all experiments. The results showed that β1 -AR mAbs induced premature ventricular contractions in guinea-pigs and isolated guinea-pig hearts. In addition, β1 -AR mAbs decreased the threshold of VT/VF and prolonged the duration of VT/VF. Furthermore, β1 -AR mAbs shortened the corrected QT interval and effective refractory period, and prolonged late-phase repolarization of the monophasic action potential (MAPD90-30 ). These changes in electrophysiological parameters might be attributed, at least in part, to the arrhythmogenicity of β1 -AR mAbs.

To observe the effect of moxibustion therapy on heme oxygenase-1 (HO-1) and monocyte chemoattractant protein-3 (MCP-3) protein expressions in the colonic mucosa of rats with Crohn’s disease (CD), and to explore the intestinal mucosal immune mechanism of moxibustion therapy in treating CD. The CD rat model was established using the internationally accepted Morris method. The rats were randomly divided into a model group, a herbal cake-partitioned moxibustion group, a mild moxibustion group, a cigarette moxibustion group and a hot compress group, which were compared with the normal group. Except the normal group and the model group, rats in the other groups accepted different moxibustion therapies on bilateral Tianshu (ST 25). Hematoxylin-eosin (HE) staining was conducted and the pathological changes of the colon were observed under light microscope; the expressions of HO-1 and MCP-3 protein in rat’s colonic mucosa were determined by immunohisto-chemistry. Compared with the normal group, rats in the model group showed mucosal defect, villus destruction or loss, submucosal congestion and edema, glandular destruction or disappearance, reduced goblet cells, ulcer formation, significantly increased positive target area and positive target integral optical density of HO-1 and MCP-3 protein expression (all P

Background Cytomegalovirus (CMV) infection is deemed to be a major cause of morbidity and mortality in patients after kidney transplantation. The purpose of this study was to analyze the incidence of CMV infection and risk factors for CMV infection in our center, to help in determination of its impact on the kidney function in this patient population, and to provide new ideas for the prevention and treatment of CMV infection. Methods A total of 319 kidney transplant recipients from our center were studied between January 2000 and December 2015. The CMV viral load in each kidney transplant patients was monitored with the use of CMV quantitative nucleic acid testing (CMV-QNAT). Laboratory data and other medical records were also collected. Results The incidence of CMV infection was 8.8% in our studied patients. The patients within 3 to 6 months and 5 to 10 years after transplantation had a higher risk of CMV infection. CMV infection was probably correlated with lower white blood cell counts but elevated hemoglobin, serum creatinine, blood urea nitrogen, potassium, and estimated glomerular filtration rate (eGFR). Anti-CMV immunoglobulin (Ig)G and history of allograft rejection were also associated with CMV infection. In multivariate regression analysis, white blood cells, eGFR, anti-CMV IgG, and history of allograft rejection were the independent risk factors associated with CMV infection in kidney transplantation patients. Conclusions CMV infection was an important complication after kidney transplantation, particularly in these patients with allograft impairment.

Objective To investigate the influence of moxibustion products on mitochondrial transmembrane potential (MTP) and mRNA expression of Bax/Bcl-2 in alveolar type II epithelial A549 cells, and to further explore influence of moxibustion products on the oxidative damage of A549 cells.

Objective To observe the effect of moxibustion on learning and memory abilities, corticosterone and glucocorticoid receptor (GR) in subacute aging rats.

Objective To observe the clinical effect and syndrome scores improvements of herbal cake-partitioned moxibustion (HPM) and ginger-partitioned moxibustion (GPM) in treating ulcerative colitis (UC).

To investigate the efficacy and mechanisms of moxibustion-based treatment of chronic gastritis (CG), and to provide an objective basis for treating CG using moxibustion. A total of 61 CG patients were divided into an herbal cake-partitioned moxibustion group and a mild-warm moxibustion group. In both treatment groups, bilateral Tianshu (ST 25), Zhongwan (CV 12) and Qihai (CV 6) were selected for moxibustion. Before and after treatment, all the enrolled patients’ gastrointestinal disease-related traditional Chinese medicine (TCM) syndrome scores and visual analog scale (VAS) scores were measured, and the changes in the serum levels of the brain-gut peptides ghrelin, somatostatin (SS) and motilin (MTL) were observed. There was no statistically significant difference between the two groups in the clinical efficacy rate (P>0.05). After treatment, the gastrointestinal disease-related TCM syndrome scores and VAS scores were reduced to varying extents in both groups, the intra-group differences were statistically significant (all P

Objective To assess the effects of direct moxibustion on 24-hour ambulatory blood pressure (ABP) and clinical symptoms of traditional Chinese medicine (TCM) in elderly patients with essential hypertension, and to explore the antihypertensive effect and influencing factors of moxibustion.

To observe the effect of electroacupuncture (EA) and herbal cake-partitioned moxibustion on anxiety and depression in patients with Crohn’s disease (CD) in remission. Sixty CD cases were randomly allocated into an EA group (n=30) and an herbal cake-partitioned moxibustion group (n=30) using the random number table by the ratio of 1:1. In addition, 30 healthy subjects were included in a control group. Bilateral Tianshu (ST 25), Qihai (CV 6) and Zhongwan (CV 12) were used in the EA and herbal cakepartitioned moxibustion groups. The treatment was done 3 times a week, for a total of 12 weeks. The efficacy was evaluated using self-rating anxiety scale (SAS), self-rating depression scale (SDS) and traditional Chinese medicine (TCM) symptom scores. Before treatment, the SAS and SDS scores in CD patients were remarkably higher than those in healthy subjects. After EA or herbal cake-partitioned moxibustion treatment, the SAS and SDS scores were significantly decreased in both groups, showing significant intra-group differences (P 0.05). Both EA and herbal cake-partitioned moxibustion can significantly decrease abnormally high SAS and SDS scores in CD patients as well as TCM symptom scores. The two therapies share similar effects in alleviating common symptoms and improving anxiety and depression.

Objective To investigate the effects of different directions of moxibustion therapy on hemorheology in rat models with blood stasis due to cold retention.

Background. Clinical studies suggest that acupuncture and moxibustion therapy in ulcerative colitis (UC) can regulate bowel inflammation, and these treatments have the advantages of low rates of adverse reactions and recurrence as well as good long-term efficacy. We reviewed the current status of clinical studies of the treatment. Methods. Randomized controlled trials (RCTs) using the therapy as the major intervention for treating UC were included from 1995 to 2015. The extracted data mainly included diagnostic standards, treatment methods, selection of acupoints, treatment times and courses, and efficacy determination criteria. Results. The use of diagnostic standards and efficacy criteria lacked unification and standardization. There were two main groups: acupuncture and moxibustion therapy combined with drug treatment and the use of all types of acupuncture and moxibustion therapy alone or in combination. The acupoint compositions included distal-proximal point combinations, back-shu point and front-mu point combinations, and acupuncture through meridians. The treatment courses in all the clinical trials had large variations. Conclusion. The treatment of UC in the examined articles was mainly based on the classical theory. However, many links of the clinical regimen design were still lacking, which affected the repeatability of the clinical studies and the accuracy of the clinical conclusions.

Blood glucose is of great importance to development and metabolic homeostasis in fetuses. Stimulation of harmful factors during gestation induces pathoglycemia. Angiotensin II type 1 receptor autoantibody (AT1-AA), a newly discovered gestational harmful factor, has been shown to induce intrauterine growth restriction in fetuses and glucose disorders in adults. However, whether and how AT1-AA influences the blood glucose level of fetuses during gestation is not yet clear. The purpose of the current study was to observe the fetal blood glucose level of AT1-AA-positive pregnant rats during late pregnancy and to determine the roles that hepatic glucose transporters play in this process. We established AT1-AA-positive pregnant rats by injecting AT1-AA into the caudal veins of rats in the 2nd trimester of gestation. Although the fetal blood glucose level in the 3rd trimester of gestation decreased, hepatic glucose uptake increased detected. Through separating membrane and cytosolic proteins, we demonstrated that both the expression and membrane transport ratio of glucose transporter 1 (GLUT1), which is responsible for glucose transport in fetal hepatocytes, were upregulated, accompanied by increased expression of N-glycosyltransferase STT3A, which contributes to the N-glycosylation of GLUT1. In vitro, we identified that AT1-AA increased glucose uptake, the expression and membrane transport ratio of GLUT1 and the expression of STT3A in HepG2 cell lines via separating membrane and cytosolic proteins and immunofluorescence, resulting in the decreased glucose content in the medium. The GLUT1 inhibitor WZB117 reversed the decreases in glucose content in the medium, the increases in glucose uptake, the increases in the expression and membrane transport ratio of GLUT1 caused by AT1-AA. The N-glycosyltransferase inhibitor NGI as well as si-STT3A reversed the AT1-AA-induced upregulation of the STT3A-GLUT1-glucose uptake effect. This study demonstrates that AT1-AA lowers the blood glucose level of fetuses via the STT3A-GLUT1-glucose uptake axis in liver.

Aims: The angiotensin II type 1 receptor autoantibody (AT1-AA) leads pathological sustainable vasoconstriction. Large-conductance Ca 2+ -and voltage-activated potassium (BK) channel, mainly consist by functional α subunits and modulatory β1 subunits in vascular smooth muscle cells (SMCs), plays a vital role on vascular relaxation. Recently, it has been found that AT1-AA inhibited BK channel’s function. However, the specific mechanisms remain unclear. In this study, we further investigated the open probability, Ca 2+ sensitivity and β1 subunits of BK channel after AT1-AA stimulation. Methods and Results: BKα subunit has several splicing variants. Immunofluorescence and PCR results showed that BK channels widely distributed among mesenteric artery (MA), most of them belong to zero-type. SMCs from MA were isolated freshly and used for patch clamp within 6 hours. AT1-AA downregulated the open probability of BK channel to 57.41±21.36% (the corresponding data of baseline was considered as 100%), specifically reduced the number of simultaneous opened channels (level 4 to level 2) instead of dwell time (T 1 =0.45 ms, T 2 = 3.39 ms to T 1 =0.58 ms, T 2 =4.99 ms) and amplitude (187.4 ±16.23 to 187.3 ±13.80 pS). This phenomenon could be reversed by BK channel agonist (NS1619) (enhanced to 124%). Noteworthy, different from Ang II, AT1-AA suppressed BK channel’s open probability consistently even after 15min (Ang II: recovered to 164.8%; AT1-AA, kept at 66.0%). The activated effect of Ca 2+ on BK channel nearly disappeared after bathing with AT1-AA in inside-out recording mode (vehicle: 100% to 471.6%; AT1-AA: 100% to 163.9%). Furthermore, to explore whether AT1-AA’s inhibitory effect on BK channel was independent from β1 subunit, only ZERO-BKα-GFP subunits were transfected into HEK293T cells. AT1-AA still diminished BK channels’ function to 57.35±5.4%. However, the inhibition of AT1-AA on BK channels was almost non-existent (to 92.1±12.81%) in AT1 receptor knock out rats. Conclusion: These results demonstrated that AT1-AA sustainably decreased open probability and Ca 2+ sensitivity of BK channels even if β1 subunit were absence. This effect was AT1 receptor dependent. It remains us that BK channel may become a novel target for improving AT1-AA related hypertension.

Introduction: Angiotensin II type 1 receptor autoantibody (AT1-AA) can continuously activate angiotensin II type 1 receptor (AT 1 R), which is related to cardiovascular disease, but the exact mechanism remains obscure. The activated AT 1 R signaling is adjusted or terminated effectively via the internalization, which is mediated by β-arrestin1/2 dependent-endocytic pathways. The hypothesis is that AT1-AA can attenuate β-arrestin1/2 recruitment, leading to the decreased AT 1 R internalization and the sustained AT 1 R activation. Methods and Results: Firstly, subcellular protein fractionation and Total Internal Reflection Fluorescence were used to examine the AT 1 R internalization induced by AT1-AA. The results confirmed that AT1-AA limited the AT 1 R internalization. Then, by co-expressing YFP-labeled AT 1 R and RFP-labeled β-arrestin1/2 in HEK293 cells with different stimulant for 10 min, the image showed that AT 1 R and β-arrestin1/2 were co-localized in the cytoplasm with Ang II. However, there were rare co-localization with AT1-AA. To further investigate the AT1-AA-induced recruiting of β-arrestin1/2, we recorded the relative change of BRET ratio of the interaction between YFP-labeled AT 1 R with Rluc-labeled β-arrestin1/2. The relative change of BRET ratio was significantly elevated with the time [(the BRET ratio with Ang II for 10 min were considered as 100%; 0 (β-arrestin1/2: 0.33 ±0.58/0.00 ±1.46), 2 (89.10 ±19.36/69.27 ±4.47), 5 (107.08 ±12.17/89.27 ±1.46), 10 (100.22 ±0.38/100.00 ±13.12), 20 (93.45 ±11.59/102.93 ±10.38) and 30 min (101.45 ±16.93/114.15±8.15)] and with the concentration (EC50 β-arrestin1/2 =6.69 ±4.09/6.34 ±2.29 nmol/L) after Ang II treatment. However, AT1-AA could not induce the recruitment of β-arrestin1/2. Furthermore, pre-incubation with an inhibitor of β-arrestin1/2 dependent-endocytic pathways prolonged the duration of AT 1 R downstream PKC, ERK1/2 phosphorylation, elevate intracellular Ca 2+ and vasoconstriction caused by Ang II, which simulated AT1-AA-caused persistent AT 1 R activation. Conclusions: Our data suggested that reduced AT 1 R internalization caused by AT1-AA was attributed to the inhibition of β-arrestin1/2 recruitment, which played a key role in how AT1-AA prolonged receptor activation.

Vascular aging predisposes the elderly to the progression of many aging-related vascular disorders and leads to deterioration of cardiovascular diseases (CVD). However, the underlying mechanisms have not been clearly elucidated. Agonistic autoantibodies against angiotensin II type 1 (AT1) receptor (AT1-AAs) have been demonstrated to be pro-inflammatory and contribute to the progression of atherosclerosis. However, the association between AT1-AAs and vascular aging has not been defined. Peripheral arterial disease (PAD) is an acknowledged vascular aging-related disease. In this study, AT1-AAs were detected in the sera of patients with PAD and the positive rate was 44.44% (n=63) vs. 17.46% in non-PAD volunteers (n=63). In addition, case-control analysis showed that AT1-AAs level was positively correlated with PAD. To reveal the causal relationship between AT1-AAs and vascular aging, an AT1-AAs-positive rat model was established by active immunization. The carotid pulse wave velocity was higher, and the aortic endothelium-dependent vasodilatation was attenuated significantly in the immunized rats. Morphological staining showed thickening of the aortic wall. Histological examination showed that levels of the senescent markers were increased in the aortic tissue, mostly located at the endothelium. In addition, purified AT1-AAs-IgGs from both the immunized rats and PAD patients induced premature senescence in cultured human umbilical vein endothelial cells. These effects were significantly blocked by the AT1 receptor blocker. Taken together, our study demonstrates that AT1-AAs contribute to the progression of vascular aging and induce EC senescence through AT1 receptor. AT1-AA is a novel biomarker of vascular aging and aging-related CVD that acts to accelerate EC senescence.