Post-amputation pain causes great suffering to amputees, but still no effective drugs are available due to its elusive mechanisms. Our previous clinical studies found that surgical removal or radiofrequency treatment of the neuroma at the axotomized nerve stump effectively relieves the phantom pain afflicting patients after amputation. This indicated an essential role of the residual nerve stump in the formation of chronic post-amputation pain (CPAP). However, the molecular mechanism by which the residual nerve stump or neuroma is involved and regulates CPAP is still a mystery. In this study, we found that nociceptors expressed the mechanosensitive ion channel TMEM63A and macrophages infiltrated into the dorsal root ganglion (DRG) neurons worked synergistically to promote CPAP. Histology and qRT-PCR showed that TMEM63A was mainly expressed in mechanical pain-producing non-peptidergic nociceptors in the DRG, and the expression of TMEM63A increased significantly both in the neuroma from amputated patients and the DRG in a mouse model of tibial nerve transfer (TNT). Behavioral tests showed that the mechanical, heat, and cold sensitivity were not affected in the Tmem63a^-/- mice in the naïve state, suggesting the basal pain was not affected. In the inflammatory and post-amputation state, the mechanical allodynia but not the heat hyperalgesia or cold allodynia was significantly decreased in Tmem63a^-/- mice. Further study showed that there was severe neuronal injury and macrophage infiltration in the DRG, tibial nerve, residual stump, and the neuroma-like structure of the TNT mouse model, Consistent with this, expression of the pro-inflammatory cytokines TNF-α, IL-6, and IL-1β all increased dramatically in the DRG. Interestingly, the deletion of Tmem63a significantly reduced the macrophage infiltration in the DRG but not in the tibial nerve stump. Furthermore, the ablation of macrophages significantly reduced both the expression of Tmem63a and the mechanical allodynia in the TNT mouse model, indicating an interaction between nociceptors and macrophages, and that these two factors gang up together to regulate the formation of CPAP. This provides a new insight into the mechanisms underlying CPAP and potential drug targets its treatment.
Animals ; Mice ; Amputation, Surgical ; Chronic Pain/pathology* ; Disease Models, Animal ; Ganglia, Spinal/pathology* ; Hyperalgesia/etiology* ; Ion Channels/metabolism* ; Macrophages ; Neuroma/pathology*

ObjectiveTo investigate the association between serum 25-hydroxy vitamin D [25(OH)D] and the occurrence and outcome of stroke-associated pneumonia (SAP) in patients with acute ischemic stroke (AIS) in emergency ward. MethodsThe clinical data of 256 patients with AIS from January, 2019 to December, 2021 were collected in the emergency department of Beijing Bo'ai Hospital. Blood routine, biochemical indicators and serum concentration of 25(OH)D were detected within 24 hours after enrollment; meanwhile, National Institute of Health Stroke Scale (NIHSS) and A2DS2 score were evaluated. The patients were divided into non-SAP group (n = 164) and SAP group (n = 92) according to whether pneumonia occurred during hospitalization. Multivariable logistic regression model was used to analyze the influencing factors of SAP. The predictive ability of serum 25(OH)D and A2DS2 for SAP were evaluated by receiver operating characteristic (ROC) curves. The 28-day survival of patients with SAP was followed up. Multivariable Cox proportional hazard regression model was used to investigate the association between vitamin D nutritional status and 28-day all-cause mortality. ResultsSerum 25(OH)D was significantly lower in the SAP group than that in the non-SAP group (Z = 6.896, P < 0.001). After adjusting age, sex, infarct volume, A2DS2 score and other factors, lower serum 25(OH)D level (OR = 0.934, 95%CI 0.884 to 0.986, P = 0.014) was an independent risk factor for SAP. The areas under curve (95%CI) of serum 25(OH)D, A2DS2 score and their combined model for predicting SAP were 0.774 (0.718 to 0.824), 0.832 (0.781 to 0.876) and 0.851 (0.802 to 0.893) (P < 0.001), respectively; and the optimum cut-off values were 25(OH)D < 10.2 ng/mL, A2DS2 score > 5 points, combined prediction > 0.207, and the Youden index were 0.493, 0.662 and 0.616, respectively. A2DS2 score could improve the prediction efficiency of serum 25(OH)D (Z = 2.106, P = 0.035). After adjusting age, sex, infarct volume and NIHSS score, vitamin D deficiency was an independent risk factor for all-cause mortality after 28 days of SAP (HR = 2.871, 95%CI 1.004 to 8.208, P = 0.049) . ConclusionSerum 25(OH)D is independently associated with the occurrence and outcome of SAP in patients with AIS in emergency ward, which could serve as an independent predictor for SAP.

Postnatal heart maturation is the basis of normal cardiac function and provides critical insights into heart repair and regenerative medicine. While static snapshots of the maturing heart have provided much insight into its molecular signatures, few key events during postnatal cardiomyocyte maturation have been uncovered. Here, we report that cardiomyocytes (CMs) experience epigenetic and transcriptional decline of cardiac gene expression immediately after birth, leading to a transition state of CMs at postnatal day 7 (P7) that was essential for CM subtype specification during heart maturation. Large-scale single-cell analysis and genetic lineage tracing confirm the presence of transition state CMs at P7 bridging immature state and mature states. Silencing of key transcription factor JUN in P1-hearts significantly repressed CM transition, resulting in perturbed CM subtype proportions and reduced cardiac function in mature hearts. In addition, transplantation of P7-CMs into infarcted hearts exhibited cardiac repair potential superior to P1-CMs. Collectively, our data uncover CM state transition as a key event in postnatal heart maturation, which not only provides insights into molecular foundations of heart maturation, but also opens an avenue for manipulation of cardiomyocyte fate in disease and regenerative medicine.
Gene Expression Regulation ; Heart ; Myocytes, Cardiac/metabolism* ; Single-Cell Analysis ; Transcription Factors/metabolism*

The dysregulation of transcription factors is widely associated with tumorigenesis. As the most well-defined transcription factor in multiple types of cancer, c-Myc can transform cells by transactivating various downstream genes. Given that there is no effective way to directly inhibit c-Myc, c-Myc targeting strategies hold great potential for cancer therapy. In this study, we found that WSB1, which has a highly positive correlation with c-Myc in 10 cancer cell lines and clinical samples, is a direct target gene of c-Myc, and can positively regulate c-Myc expression, which forms a feedforward circuit promoting cancer development. RNA sequencing results from Bel-7402 cells confirmed that WSB1 promoted c-Myc expression through the β-catenin pathway. Mechanistically, WSB1 affected β-catenin destruction complex-PPP2CA assembly and E3 ubiquitin ligase adaptor β-TRCP recruitment, which inhibited the ubiquitination of β-catenin and transactivated c-Myc. Of interest, the effect of WSB1 on c-Myc was independent of its E3 ligase activity. Moreover, overexpressing WSB1 in the Bel-7402 xenograft model could further strengthen the tumor-driven effect of c-Myc overexpression. Thus, our findings revealed a novel mechanism involved in tumorigenesis in which the WSB1/c-Myc feedforward circuit played an essential role, highlighting a potential c-Myc intervention strategy in cancer treatment.

To construct an intelligent appointment register platform for clinical examination in large-scale hospitals can improve the utilization efficiency of medical resources and shorten the waiting period of patients for examination. In this study, a knowledge base of medical examination information was built through the investigation of general hospital, and all examination systems and items were integrated into the appointment register platform, so that all examinations were appointed based on the principle of completing all tests in the shortest time, thus building a unified intelligent appointment register platform for the hospital. From that, the average appointment register procedure of examinations has been shorten after launching the intelligent platform, labor costs of appointment register center and inpatient legwork have been saved, all examinations of inpatients can be finished in 48 hours, and patient satisfaction has been improved obviously.

Objective:To systematically review the methodology in clinical trial-based health economics study with cost-effectiveness ratio for nutritional drug.Methods:The literature on health economics study for nutritional drug was retrieved from PubMed and Wanfang Medical Network by October 2019. The literature was selected according to inclusion and exclusion criteria, and was assessed using the Cochrane Risk Bias Assessment Tool and Newcastle-Ottawa Scale. Its methodology such as participants and grouping, confounding factors, research perspective, cost accounting, health outcomes and health economics analysis methods, sensitivity analysis, etc, was systematically reviewed as well.Results:Four target literatures were included in this study. The participants were from gastroenterology, gastrointestinal surgery, etc. Random grouping, regression, propensity score matching method, etc, were used to control confounding factors. The research perspective needed to be clear according to the principle of health economics study. The present literatures focused on "direct medical costs" , and calculated cost-effectiveness ratio or incremental cost-effectiveness ratio to evaluate the economics of medical interventions.Conclusion:The evidence of high-quality health economics research in parenteral and enteral nutrition area in China needs to be promoted, especially in the control of confounding factors, the choice of research perspective and sensitivity analysis, which are supposed to be explored by multidisciplinary research teams in practice.

To establish the experts consensus on the management of delirium in critically ill patients.A special committee was set up by 15 experts from the Chinese Critical Hypothermia-Sedation Therapy Study Group.Each statement was assessed based on the GRADE (Grading of Recommendations Assessment,Development,and Evaluation) principle.Then the Delphi method was adopted by 36 experts to reassess all the statements.(1) Delirium is not only a mental change,but also a clinical syndrome with multiple pathophysiological changes.(2) Delirium is a form of disturbance of consciousness and a manifestation of abnormal brain function.(3) Pain is a common cause of delirium in critically ill patients.Analgesia can reduce the occurrence and development of delirium.(4) Anxiety or depression are important factors for delirium in critically ill patients.(5) The correlation between sedative and analgesic drugs and delirium is uncertain.(6) Pay attention to the relationship between delirium and withdrawal reactions.(7) Pay attention to the relationship between delirium and drug dependence/ withdrawal reactions.(8) Sleep disruption can induce delirium.(9) We should be vigilant against potential risk factors for persistent or recurrent delirium.(10) Critically illness related delirium can affect the diagnosis and treatment of primary diseases,and can also be alleviated with the improvement of primary diseases.(11) Acute change of consciousness and attention deficit are necessary for delirium diagnosis.(12) The combined assessment of confusion assessment method for the intensive care unit and intensive care delirium screening checklist can improve the sensitivity of delirium,especially subclinical delirium.(13) Early identification and intervention of subclinical delirium can reduce its risk of clinical delirium.(14) Daily assessment is helpful for early detection of delirium.(15) Hopoactive delirium and mixed delirium are common and should be emphasized.(16) Delirium may be accompanied by changes in electroencephalogram.Bedside electroencephalogram monitoring should be used in the ICU if conditions warrant.(17) Pay attention to differential diagnosis of delirium and dementia/depression.(18) Pay attention to the role of rapid delirium screening method in delirium management.(19) Assessment of the severity of delirium is an essential part of the diagnosis of delirium.(20) The key to the management of delirium is etiological treatment.(21) Improving environmental factors and making patient comfort can help reduce delirium.(22) Early exercise can reduce the incidence of delirium and shorten the duration of delirium.(23) Communication with patients should be emphasized and strengthened.Family members participation can help reduce the incidence of delirium and promote the recovery of delirium.(24) Pay attention to the role of sleep management in the prevention and treatment of delirium.(25) Dexmedetomidine can shorten the duration of hyperactive delirium or prevent delirium.(26) When using antipsychotics to treat delirium,we should be alert to its effect on the heart rhythm.(27) Delirium management should pay attention to brain functional exercise.(28) Compared with non-critically illness related delirium,the relief of critically illness related delirium will not accomplished at one stroke.(29) Multiple management strategies such as ABCDEF,eCASH and ESCAPE are helpful to prevent and treat delirium and improve the prognosis of critically ill patients.(30) Shortening the duration of delirium can reduce the occurrence of long-term cognitive impairment.(31) Multidisciplinary cooperation and continuous quality improvement can improve delirium management.Consensus can promote delirium management in critically ill patients,optimize analgesia and sedation therapy,and even affect prognosis.

To explore the effect of adipose-derived mesenchymal stem cells (ADMSCs) on ovarian damage induced by cyclophosphamide (CTX) and its mechanism.
 Methods: ADMSCs isolated from adipose tissue of female SD rats were cultured and divided into a blank group and a CTX group (n=15 in each group). CTX (75 mg/kg) was injected intraperitoneally to establish a model of ovarian damage in rats. A total of 45 female SD rats were also divided into 3 groups: Group A (15 rats, only injected intraperitoneally with 75 mg/kg CTX diluted with 1 mL 0.9% saline), Group B [15 rats, injected intraperitoneally with 75 mg/kg CTX diluted with 1 mL 0.9% saline, after 4 estrus cycles, injected 0.6 mL ADMSCs (6×105 cells) by the tail vein], and Group C [15 rats, injected intraperitoneally with 75 mg/kg CTX diluted with 1 mL 0.9% saline, after 4 estrus cycles, injected 40 mL ADMSCs (20 mL per side, 2×104 cells) in situ ovarian]. After 4 estrus cycles, the changes of quality of life, ponderal growth were recorded, the sex hormone levels [estradiol (E2), follicle-stimulating hormone (FSH)] were tested by ELISA, and the morphology of ovarian tissue and follicle count were observed by HE staining. The expression of BMP-15, Bcl-2 and Bax in ovarian tissues were tested by immunohistochemistry, real-time PCR or Western blotting. The apoptosis rate of follicular cells was detected by TdT-mediated dUTP nick end labeling (TUNEL) assay.
 Results: After transplantation of ADMSCs, compared with the Group A, their quality of life of rats in the Group B and C was improved, and the ponderal growth was increased (both P<0.01). Compared with the Group A, the serum E2 levels in the Group B and the Group C were increased (P<0.01, P<0.05), and the FSH levels in the Group B and C were decreased (both P<0.01). The granular cell layer, the number of corpus lutein and the count of various grade follicles were significantly increased, and many new follicles and mature oocytes were observed in the Group B and C. Compared with Group A, the count of primitive follicles, sinusoidal follicles, pre-ovulation follicles and total follicles, and pre-sinusoidal follicles were dramatically increased in the Group B. The follicle at all levels count was increased in the Group C than that in the Group A (all P<0.01). Comparing with the Group A, the expressions of BMP-15 and Bcl-2 were increased (all P<0.01), the expressions of Bax was decreased (both P<0.01), and the apoptosis rates of follicular cells were decreased in the Group B and C (both P<0.01). However, there was no difference between the Group B and the Group C in the above indexes (all P>0.05).
 Conclusion: ADMSCs transplantation can effectively repair ovarian damage induced by CTX in rats, which may be achieved by inhibiting mitochondrial apoptosis of granulosa cells.
Animals ; Cyclophosphamide ; Female ; Mesenchymal Stem Cells ; Ovary ; Quality of Life ; Rats ; Rats, Sprague-Dawley

DNA methylation is a significant epigenetic modification mode, which plays an important role in embryo reprogramming, stem cell differentiation and tumor occurrence. The ten-eleven translocation (TET) enzyme is a crucial demethylation enzyme, which can catalyze 5-methylcytosine(5mC) to 5-hydroxymethylcytosine(5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine(5caC). These bases represent the epigenetic modifications of DNA and regulate the process of DNA methylation. Understanding the role of TET enzyme in regulating the DNA methylation modification and gene expression can help us to gain the knowledge for the normal growth development and epigenetic regulation in human diseases.
5-Methylcytosine ; metabolism ; Cell Differentiation ; DNA ; DNA Methylation ; DNA-Binding Proteins ; Epigenesis, Genetic ; Humans

Objective To analyze the prevalence characteristics of fatty liver in young and middle-aged people,and to explore the risk factors of the disease,so as to provide clinical evidence for the prevention and treatment of fatty liver.Methods In 756 young and middle-aged healthy subjects with age≤50 years old,there were 197 fatty liver cases were diagnosed by ultrasonic test during 2015 year.The prevalence of different characteristics in young and middle-aged fatty liver was analyzed,the difference of blood biochemical index between fatty liver and non fatty liver group was compared,and risk factors of fatty liver was explored by binary logistic regression model.Results The total prevalence rate of fatty liver in young and middle-aged people was 26.1%(197/756),among which 33.1% (119/359) were male and 19.6%(78/397) were female,the prevalence rate of male was significantly higher than that of female(χ2=17.833,P<0.05).The prevalence rate of fatty liver increased with age(χ2=33.296,P<0.05),which in 40-50 years old was 37.1%(111/299) and significantly higher than that in 20 years-(15.0%)(24/160) and 30 years-(20.9%)(62/297).Logistic regression analysis on influencing factors of fatty liver prevalence showed that age,sex,body mass index(BMI),drinking,diabetes and fasting blood glucose(FPG),triglyceride(TG),total cholesterol(TC) were closely related to fatty liver(P<0.05),overweight,obesity,drinking,diabetes increased the risk of fatty liver disease.Biochemical indicators(FPG,TG,TC) in fatty liver group were higher than those in non fatty liver group((7.09±1.47) mmol/L vs.(5.14±1.71) mmol/L,(5.98±1.23) mmol/L vs.(4.95±1.42) mmol/L,(2.03±0.45) mmol/L vs.(1.23±0.67) mmol/L,t=271.905,98.866,278.255;P<0.05).Conclusion The prevalence rate of fatty liver in young and middle-aged people is high,and controlling body weight,give up drinking,active treating diabetes,reducing blood glucose and blood lipids can effectively decrease the prevalence of fatty liver in young and middle-aged people.