Objective:To explore the role of corticotrophin releasing factor receptor 2 (CRFR2) in regulating pain sensitization and anxiety and its mechanism in chronic migraine mice.Methods:Forty-eight C57BL/6J mice were randomly divided into control group, model group, NBI35965 group and K41498 group ( n=12); chronic migraine models in the later 3 groups were established by intraperitoneally administrating 10 mg/kg nitroglycerin on the 1 st, 3 rd, 5 th, 7 th and 9 th d; mice in the NBI35965 group and K41498 group were injected with 100 nL NBI35965 or K41498 solution into the bilateral trigeminal nucleus caudalis on the 2 nd, 4 th, 6 th and 8 th d, and mice in the control group were injected with same volume of normal saline. Von frey fiber was used to detect the orbitofrontal mechanical pain threshold 2 h after intraperitoneal injection on the 1 st, 3 rd, 5 th, 7 th and 9 th d, and at 11 a.m. on the 10 th d. Elevated plus maze was used to detect the anxiety-like behaviors at 11 a.m. on the 11 th d. Western blotting was performed to detect the protein expressions of corticotrophin releasing factor (CRF), corticotrophin releasing factor receptor 1 (CRFR1), CRFR2 in the trigeminal nucleus caudalis. Real-time quantitative PCR (RT-qPCR) was used to detect the CRFR1 and CRFR2 mRNA expressions in the trigeminal nucleus caudalis. Immunofluorescent staining was used to detect the protein expressions of calcitonin gene-related peptide (CGRP), immediate-early gene c-fos, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba-1) in the trigeminal nucleus caudalis. Results:Compared with the control group, the model group, NBI35965 group and K41498 group had significantly decreased orbitofrontal mechanical pain thresholds 3, 5, 7, 9, and 10 d after intraperitoneal injection ( P<0.05); compared with model group, the K41498 group had significantly increased orbitofrontal mechanical pain thresholds 7, 9, and 10 d after intraperitoneal injection ( P<0.05). Compared with control group, the model group, NBI35965 group and K41498 group had significantly decreased entries and shorter time in opened arms ( P<0.05); compared with the model group, the K41498 group had significantly increased entries and shorter time in opened arms ( P<0.05). Compared with the control group, the model group, NBI35965 group and K41498 group had significantly higher CRF and CRFR2 protein expressions in the trigeminal nucleus caudalis ( P<0.05); compared with the model group, the K41498 group had statistically lower CRF protein expression in the trigeminal nucleus caudalis ( P<0.05). Compared with the control group, the model group, NBI35965 group and K41498 group had significantly higher CRFR2 mRNA expression in the trigeminal nucleus caudalis ( P<0.05). Compard with the control group, the model group, NBI35965 group and K41498 group had significantly increased CGRP, c-fos, Iba-1 and GFAP protein expressions in the trigeminal nucleus caudalis ( P<0.05); compared with the model group, the K41498 group had significantly decreased CGRP and c-fos protein expressions in the trigeminal nucleus caudalis ( P<0.05). Conclusion:CRFR2 can alter the orbitofrontal pain sensitization and anxiety-like behaviors in chronic migraine mice by regulating neuronal activation and CGRP release in the trigeminal nucleus caudalis.

Objective:To document the impact of information-based, stepwise, intensive rehabilitation therapy on patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods:Eighty such patients in an intensive care unit (ICU) were randomly divided into a control group and an observation group, each of 40. The control group received routine ICU rehabilitation, while the observation group underwent information-based, step-wise ICU rehabilitation. Upon admission to and discharge from the ICU, the forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), FEV1/FVC, and diaphragm functioning were compared between the two groups. The duration of mechanical ventilation, the incidence of ventilator-associated pneumonia (VAP), length of stay in the ICU, incidence of delirium, and the incidence of deep vein thrombosis (DVT) were also recorded. The number of patients readmitted to the ICU after discharge, and the 28-day hospital mortality rate were recorded as well.Results:Significant improvement was observed in both groups in terms of their lung and diaphragm functioning, as well as in the rehabilitation- and hospital-related indicators. At discharge, significantly greater improvements were observed in the observation group compared with the control group in terms of their average FEV1, FVC, FEV1/FVC, inspiratory and expiratory diaphragm thickness, and diaphragm thickening rate. The average duration of mechanical ventilation and of rehabilitation interruptions was significantly less in the observation group. And incidents of accidental extubation, VAP, delirium and DVT were significantly fewer in the observation group as well. Their ICU stays tended to be significantly shorter without any significant difference between the two groups in the 28-day hospital mortality rate. The control group spent significantly less time in their daily rehabilitation sessions, with the result that significantly fewer of them achieved a grading of 2 or better on the mMRC respiratory questionnaire.Conclusion:Information-based stepwise intensive rehabilitation treatment can effectively improve the pulmonary and diaphragmatic function of AECOPD patients admitted to an ICU, shorten their mechanical ventilation time and the length of their ICU stay, and lower their incidence of VAP and DVT during hospitalization.

Berberine (BBR) as one of the most effective natural products has been increasingly used to treat various chronic diseases due to its immunosuppressive/tolerogenic activities. However, it is unknown if BBR can be applied without abrogating the efforts of vaccination. Here we show that priming of CD8⁺ T cells in the presence of BBR lead to improved central memory formation (Tcm) with substantially reduced effector proliferation, primarily orchestrated through activation of AMPK and Stat5. Tcm derived from vaccinated mice fed with BBR were able to adoptively transfer protective immunity to naïve recipients. Vaccination of BBR-fed mice conferred better memory protection against infection without losing immediate effector efficacy, suggesting appreciable benefits from using BBR in vaccination. Thus, our study may help to lay the groundwork for mechanistic understanding of the immunomodulatory effects of natural products and their potential use as adjuvant that allows the design of novel vaccines with more desirable properties.

Objective:To evaluate the effect of controlled low central venous pressure with milrinone on laparoscopic hepatectomy in the patients.Methods:Fifty American Society of Anesthesiologists physical statusⅠ-Ⅲ patients of both sexes, aged 18-64 yr, with body mass index of 18-30 kg/m 2, of Child-Pugh grade A or B, undergoing elective laparoscopic hepatectomy, were divided into 2 groups ( n=25 each) using a random number table method: milrinone group (group M) and nitroglycerin group (group NG). After the start of surgery, milrinone 0.5 μg·kg -1·min -1 was continuously infused in group M, and nitroglycerin was continuously infused with the initial dose of 0.5 μg·kg -1·min -1 to maintain central venous pressure (CVP)≤5 mmHg in group NG.Mean arterial pressure and heart rate were recorded on admission to the operation room (T 0), at skin incision (T 1), at the beginning of liver resection (T 2), at completion of liver resection (T 3), at the end of operation (T 4), and CVP, cardiac index and stroke volume variation were recorded at T 1-4.Internal jugular vein blood samples were collected to determine the concentrations of hemogloblin, blood lactate at T 1 and T 4, and serum alanine aminotransferase, aspartate aminotransferase and creatinine concentrations at 1, 3 and 7 days after surgery.The score of blood oozing in hepatic surgical field, amount of norepinephrine used, blood loss, postoperative recovery and occurrence of complications within 7 days after operation were recorded. Results:Compared with group NG, cardiac index was significantly increased at T 2, 3, the CVP was decreased at T 2, the blood oozing score, blood loss, consumption of norepinephrine, and concentrations of blood lactate were decreased, and the postoperative drainage indwelling time was shortened in group M ( P<0.05). There was no significant difference in the serum alanine aminotransferase, aspartate aminotransferase and creatinine concentrations and incidence of postoperative complications at 1, 3 and 7 days after operation between the two groups ( P>0.05). Conclusions:Milrinone is better than nitroglycerin in decreasing central venous pressure, reducing blood loss, maintaining stable circulatory function and tissue perfusion in laparoscopic hepatectomy.

Objective:To explore any effect of repeated transcranial magnetic stimulation (rTMS) on the recovery of neurological functioning and the expression of NOD-like receptor family pyrin domain containing 3 (NLRP3) and inflammatory factors after ischemic stroke.Methods:Sixty-four C57BL/6J mice were randomly divided into a normal control group, a model group, a sham stimulation group and an observation group, each of 16. All mice except those of the normal control group received middle cerebral artery occlusion using the suture method to model an ischemic stroke. After the modeling the observation group was given 1Hz rTMS daily for 7 consecutive days, while the sham stimulation group was given sham rTMS. After the intervention, Zea-Longa scores were used for all of the groups, and the size of the cerebral infarct was measured using triphenyltetrazolium chloride staining. The expression of NLRP3 around the cerebral infarction was detected using immunofluorescence, while that in the brain tissue was measured using Western blotting. The expression of interleukin-1β and IL-18 in the brain tissue was detected using enzyme-linked immunosorbent assays.Results:Compared with the normal control group, a significant increase was observed in the other groups′ average neurological function impairment scores. Expression of NLRP3, IL-1β and IL-18 in the model and sham stimulation groups also increased, with large cerebral infarcts in the cortex and hippocampus. Compared with the sham stimulation and model groups, there was a significant decrease in the average neurological dysfunction scores, the area of cerebral infarction in the cortex and hippocampus, as well as the expression of NLRP3, IL-1β and IL-18 in the observation group.Conclusions:Low-frequency rTMS can promote the recovery of damaged nerve function after an ischemic stroke, at least in mice. It can reduce the size of cerebral infarction, and inhibit neuronal pyroptosis, which is closely related to the down-regulation of NLRP3, IL-1β and IL-18 expression.

Objective:To investigate the prognoses of pulmonary adenocarcinoma patients with leptomeningeal metastases (LM) and explore their influencing factors.Methods:A retrospective analysis was performed. The clinical data, imaging features and treatment plans of pulmonary adenocarcinoma patients with LM admitted to our hospital from January 2010 to June 2021 were collected. Overall survival (OS) was used as the prognostic evaluation criterion and patients were divided into good prognosis group (OS≥6 months) and poor prognosis group (OS<6 months) accordingly. Logistic regression analysis was used to evaluate the influencing factors for prognoses of pulmonary adenocarcinoma patients with LM. These patients were grouped according to different Karnofsky performance status (KPS) scores and different treatment methods, and survival curves were drawn to compare their OS.Results:A total of 173 pulmonary adenocarcinoma patients with LM were enrolled in the study, including 75 with good prognosis and 87 with poor prognosis. There were significant differences in the KPS scores, pulmonary adenocarcinoma lesion controlled status, giving third generation tyrosine kinase inhibitor (TKI) therapy or not, giving systemic chemotherapy and/or whole brain radiotherapy or not between the two groups ( P<0.05). Multivariate Logistic regression analysis showed that KPS scores and pulmonary adenocarcinoma lesion controlled status were independent influencing factors for prognoses ( OR=4.186, 95%CI: 1.583-11.070, P=0.004; OR=4.198, 95%CI: 1.499-11.760, P=0.006). Survival curves showed median OS of 8.2 months for all patients ( 95%CI: 6.5-9.8). The OS in patients with low-risk(KPS scores≥60) was significantly higher than that in patients with high-risk(KPS scores<60), that in patients accepted TKI treatment was significantly higher than that in patients not accepted TKI treatment, and that in patients accepted TKI and systemic chemotherapy was significantly higher than that in patients accepted TKI alone ( P<0.05). Conclusion:Patients with high KPS scores and controlled pulmonary adenocarcinoma can have relatively good prognosis; TKI treatment and combination therapy may prolong OS of these patients.

Objective To investigate the expression of Scinderin(SCIN myoprotein) in breast cancer tissues and adjacent tissues,and to explore the relationship between the expression of Scinderin and different molecular subtypes of breast cancer as well as the clinical factors.Methods Immunohistochemical staining method was used to detect the expression of SCIN in 120 cases of breast carcinoma and 30 adjacent tissues.The relation between SCIN expression in breast cancer tissue and molecular subtypes,pathologic stage,age,tumor size,lymph node metastasis was analyzed.Results SCIN expression level in breast cancer tissue was lower than in the tissues adjacent to carcinoma (6.06±3.32 vs 7.77±3.32,P＜0.05).SCIN expression was associated with breast cancer molecular subtypes (P＜0.05),and it was irrelevant with age,tumor size,histological grade,lymph node metastasis,or clinical stage (P＞0.05).Conclusions The expression of SCIN in breast cancer tissues was lower than in the adjacent tissues.It is associated with breast cancer molecular subtypes.SCIN could become the protein markers of breast cancer molecular targeted therapy.

Objective:To explore the role of Pim-1 in the pathology of inflammatory bowel disease and the potential effect of Pim-1 inhibitor on treating such disease.Methods:Forty-five BALB/c mice were randomly divided into 5 groups (n=9):A normal control group,a inflammatory bowel disease group,two different dose of Pim-1 inhibitor treatment groups,and steroidhormone treatment group.The model of inflammatory bowel disease was induced by intracolonic administration of 2,4,6-trinitrobenzenestdfonic acid (TNBS) and ethanol mixture.Mice were treated with Pim-1 inhibitor [intraperitoneal inject,5 or 10 mg/(kg.d)] for 5 days and prednisone (intragastric administration,0.1 mg/d) for 5 days.The DAI,colon length,gross score and pathological grade were evaluated.The expressions ofT cell master transcription factors T-box expressed in T cells (T-bet),GATA binding protein 3 (GATA-3),RA orphan receptorγ (RORyt)and forkhead box P3 (Foxp3) were measured by Real-time PCR and Western blot,respectively.Results:Pim-1 inhibitor and prednisone showed therapeutic effect on acute TNBS colitis in vivo.GATA3 and RORγt were significantly up-regulated in acute TNBS colitis (P＜0.05).In contrast,the expression of Foxp3 was suppressed in the inflammatory bowel disease group,whereas it did not cause any significant change in T-bet expression (P＞0.05).Administration of Pim-1 inhibitor and prednisone resulted in suppression of GATA3,RORγt expression,and the increase of Foxp3 expression (P＜0.05).Administration of Pim-1 inhibitor and prednisone resulted in inhibition of T-bet mRNA expression (P＜0.05),but only prednisone could inhibit T-bet protein expression (P＞0.05).Conclusion:Pim-1 inhibitor significantly suppresses Th2-and Th17-type immune responses.Furthermore,Pim-1 inhibitor could induce T-cell differentiation towards a Treg phenotype.Pim-1 inhibitor has therapeutic effect on acute TNBS colitis.

Objective To explore the application of nanodisc in functional and drug discovery research of G protein-coupled receptor (GPCR).Methods The purified recombinant 5-Hydroxytryptamine 2B receptor (5-HT2BR) was reconstituted into nanodisc complex.Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclution chromatography were performed to evaluate the reconstitution reaction,followed by the use of surface plasmon resonance to validate the ligand-binding activity of 5-HT2BR after reconstitution.Results 5-HT2B R was effectively self-assembled into nanodisc while maintained its binding activity toward the antagonist SB204741.Conclusions The presented study provided potential application of 5-HT2B R-nanodisc for the development of subtype-selective drugs against 5-HT2B R and the fundamental of utilizing nanodisc for GPCR structural and functional studies as well as drug discovery.

Objective To investigate the risk factors of thrombopenia(TP)in septic patients complicated with acute kidney injury (AKI).Methods Two hundred and sixty five septic patients complicated with AKI admitted in Intensive Care Unit ICU of Zhejiang Provincial People''s Hospital during January 2012 and December 2016 were enrolled in the study.The clinical data, results of laboratory tests, Acute Physiology and Chronic Health Evaluation (APACHEII) scores, Sequential Organ Failure Assessment (SOFA) scores, therapeutic intervention, and 28-day mortality were documented.Among 265 patients, TP occurred within 7 days in 112 cases (TP group) and did not occur in 153 cases (non-TP group).Multivariable Logistic regression analysis was performed to analyze the risk factors of TP.Results The 28-day mortality rate in TP group was higher in TP group than that in non-TP group (47.3% vs.33.3%, χ2=5.307,P<0.05).Univariate analysis showed that age, C-reactive protein (CRP), procalcitonin (PCT) and APACHEII score, SOFA score, continuous renal replacement therapy (CRRT), heparin anticoagulation, shock, usage of linezolid and bloodstream infections were associated with TP in septic patients with AKI(all P<0.05).Multivariable Logistic regression analysis showed that age≥65 (OR=4.53, 95%CI 1.23-9.24,P<0.05), CRRT(OR=5.24,95%CI 2.14-14.56,P<0.01), heparin anticoagulation(OR=4.56,95%CI 2.13-8.46,P<0.01), usage of linezolid(OR=2.35,95%CI 1.25-5.24,P<0.01), shock(OR=2.15,95%CI 1.03-4.96,P<0.01)and bloodstream infections(OR=4.26,95%CI 1.36-12.48,P<0.01)were independent risk factors for septic patients with TP.Conclusion For septic patients with AKI having these risk factors, the platelet counts should be closely monitored, and intervention measures should be given to reduce the occurrence of TP.