Tryptophan metabolism plays an important role in immunometabolism in sepsis,and the expression of indoleamine 2,3-dioxygenase 1(IDO1),the key enzyme of tryptophan metabolism,is up-regulated during sepsis.This study was designed to construct IDO1 gene knockout THP-1 cell line using CRISPR/Cas9,and to provide a cell model for studying the role of IDO1 in macrophage function.Three gRNAs were designed for the IDO1 gene,and inserted into YKO-Lentiviral gRNA plasmid respectively to construct IDO1-gRNA recombinant plasmid,which then confirmed by restriction enzyme digestion and sequencing,and packaged as Lenti-IDO1-gRNA lentivirus.THP-1 cells were infected with Cas9 lentivirus to construct a THP-1 Cas9 cell line stably expressing Cas9 protein.Then THP-1 Cas9 cells were infected with the Lenti-IDO1-gRNA lentivirus for IDO1 gene knockout,and the monoclonal cell lines were screened by limited dilution method.The efficiency of IDO1 knockout was identified by T7E1 digestion test,PCR product sequencing and Western blotting.The proliferative activity of THP-1 cells was detected by CCK8 assay;the expression of CD11b,CD68 and CD 14 in THP-1 macrophages were detected by flow cytometry;and the phagocytic function of THP-1 macrophages was detected by neutral phagocytosis test.T7E1 restriction enzyme digestion results showed that all the three gRNAs could effectively edit IDO1 gene,and gRNA2 had the highest editing efficiency.Sequencing of PCR products showed that IDO1 gene was mutated in three THP-1 IDO 1-KO monoclonal cell lines,and Western blot result showed that IDO1 protein was not expressed,suggesting THP-1 IDO1-KO cells were successfully constructed.CCK-8 proliferation assay showed that IDO1 gene knockout could inhibit the proliferation of THP-1 cells.Flow cytometry showed that IDO1 gene knockout down-regulated the expression of CD1 1b and CD68 in THP-1 macrophages,while up-regulated the expression of CD 14.The results of neutral red phagocytosis test showed that IDO1 gene knockout promoted the phagocytosis of macrophages.In conclusion,the IDO1 gene knockout THP-1 cell line has successfully constructed by CRISPR-Cas9,and IDO1 plays an important role in regulating the proliferation activity and differentiation of THP-1 cells,as well as the phagocytic function of THP-1 macrophages.

It has been proposed by Basic Questions On Proper Therapies for Different Diseases Geographically （《素问·异法方宜论篇》） that "wei （痿） diseases should be treated by Daoyin （导引）". Furthermore， it is clarified that the indications of Daoyin are those conditions related to spleen and dampness caused by dampness pathogen， excessive food intake and less exercise， and mainly manifested as heavy limbs， fatigue and flaccidity， which is similar to the metabolic imbalance in the early stage of glucose or lipid metabolism disorder in modern medicine. Based on modern clinical and basic research evidence， Daoyin can inhibit the response of inflammation， alleviate oxidative stress， regulate intestinal microbiota， and modulate gene expression to improve metabolic abnormalities， and this will provide ideas for researches on the indications of Daoyin.

ObjectiveTo investigate the current status and development needs of evidence-based medicine （EBM） capability in ethnic minority medicine， and explore effective strategies to enhance EBM capability in this field. MethodsThe questionnaire survey was conducted in various ethnic minority medical institutions and research organisations. The questionnaire covered three dimensions， firstly， perceptions and attitudes towards evidence-based medicine； secondly， advantages and challenges in the development of ethnic minority medicine； thirdly， demands and recommendations for enhancing evidence-based medicine capability in ethnic minority medicine. ResultsA total of 501 valid questionnaires were collected， of which 103 questionnaires were collected by re-sending to minority medicine regions with insufficient participation. The questionnaires included 354 responses （70.66%） from practitioners of minority medicine， including Tibetan medicine， Mongolian medicine， Uyghur medicine， Zhuang medicine， and Korean medicine. Among the 501 questionnaires， 146 respondents （29.14%） indicated that they knew about EBM， 355 respondents （70.86%） had either a "general understanding" or had "not heard about" EBM before， and 469 respondents （93.61%） believed that introducing ECM could promote the development of ethnic minority medicine. The primary challenge in promoting EBM in the field of ethnic minority medicine is the lack of professionals in EBM and a lack of understanding of how to apply it into clinical practice （442 respondents， 88.22%）. In the 9-point importance rating for enhancing evidence-based abilities， high scores were achieved in standardization of clinical practice guidelines （7.50±1.90） and methods for sample sizes in clinical research （7.45±1.90）. Regarding the demand for improving clinical research literacy， expert academic lectures， and experience sharing （404 respondents， 80.64%） and evidence-based methodology monographs on ethnic minority medicine （401 respondents， 80.04%） were emphasized. ConclusionsPractitioners in ethnic minority medicine hold a positive attitude towards integrating EBM. However， there remains substantial room for the education and dissemination of EBM. Enhancing evidence-based capabilities can be achieved through specific measures such as cultivating or recruiting talents in EBM， establishing evidence-based support platforms for clinical research， organizing regular academic lectures and exchanges， and strengthening the construction of theoretical frameworks and evaluation systems tailored to ethnic minority medicine， thereby following a path of evidence-based practices aligned with the unique characteristics of ethnic minority medicine.

This article reported a case of neurodevelopmental disorder accompanied by spastic diplegia and visual impairment with the manifestation of small fetal head circumference. Prenatal ultrasonography performed at 33 +5 weeks of pregnancy revealed small fetal head circumference (-2.61SD) and oligohydramnios. Whole-exome sequencing identified a heterozygous frameshift variation of c.1623_1624insA (p.R542Tfs*30) in the CTNNB1 gene (NM_001904.4) of the fetus. No phenotypic abnormalities or corresponding gene variations were detected in the parents, suggesting it was a de novo variation. Based on the clinical manifestations, the fetus was diagnosed with a neurodevelopmental disorder accompanied by spastic diplegia and visual defects. Following genetic counseling, the pregnant woman chose to terminate the pregnancy.

Diabetic neuropathic pain (DNP) is the most common disabling complication of diabetes. Emerging evidence has linked the pathogenesis of DNP to the aberrant sprouting of sensory axons into the epidermal area; however, the underlying molecular events remain poorly understood. Here we found that an axon guidance molecule, Netrin-3 (Ntn-3), was expressed in the sensory neurons of mouse dorsal root ganglia (DRGs), and downregulation of Ntn-3 expression was highly correlated with the severity of DNP in a diabetic mouse model. Genetic ablation of Ntn-3 increased the intra-epidermal sprouting of sensory axons and worsened the DNP in diabetic mice. In contrast, the elevation of Ntn-3 levels in DRGs significantly inhibited the intra-epidermal axon sprouting and alleviated DNP in diabetic mice. In conclusion, our studies identified Ntn-3 as an important regulator of DNP pathogenesis by gating the aberrant sprouting of sensory axons, indicating that Ntn-3 is a potential druggable target for DNP treatment.
Mice ; Animals ; Diabetes Mellitus, Experimental/metabolism* ; Axons/physiology* ; Diabetic Neuropathies ; Sensory Receptor Cells/metabolism* ; Neuralgia/metabolism*

Objective To discuss the application effects of emergency care path on rescue of patients with acute myocardial infarction.Methods A total of 84 patients with acute myocardial infarction were selected by purposeful sampling method as the study subjects from January 2015 to December 2016 in Emergency Department in the Second Affiliated Hospital of Qiqihar Medical University, and were divided into control group and observation group by random number table, in which 40 cases were treated with traditional nursing model in the control group, and 44 cases were treated with emergency nursing path model in the observation group. We compared Seattle angina questionnaire, nursing satisfaction scale for patients' evaluation.Results After interventions, the patients' scores of observation group were (18.03±1.55),(17.82±2.15),(18.27±1.69), (18.92±0.76),(18.95±0.57),higher than (15.46±1.76),(14.76±2.01),(15.72±2.17),(14.87±1.56), (14.82±1.27)in the control group in angina pain, restriction degree of body activity, occurrence of angina, disease recognition degree, pain control satisfaction (t=-5.663, -5.791, -5.691, -6.702, -6.772;P<0.05). After intervention, the patient's satisfaction in the observation group was 97.73%, higher than 77.50% in the control group (χ2=8.174,P<0.01).Conclusions Emergency care pathways can rescure patients with acute myocardial infarction, effectively improve the patient's symptoms, and contribute nursing satisfaction.

Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.

BACKGROUND: At present, there are considerable investigations and reports concerning the vascular remodeling cum pathology, molecular biology, immunohistochemistry, drug effect to the disease of the heart and great vessels. But varicocele is seldom reported. OBJECTIVE: To observe the change of the qualitation and quantitation of actin and collagen in varicocele vessel wall. DESIGN, TIME AND SETTING: A synchronized non-randomized control observation was carried out in the Department of Urinary Surgery, the First Affiliated Hospital of Chongqing University of Medical Sciences from March 2006 to June 2007. PARTICIPANTS: Thirty varicocele patients treated in Department of Urinary Surgery, the First Affiliated Hospital of Chongqing University of Medical Sciences from March 2006 to June 2007, were recruited for experimental group in this study. Their age was (26.29?9.12) years, and the course of disease was (2.76?1.83) years. Meanwhile, 24 normal spermatic veins from the donator of kidney transplantation, aged (39.56?8.32) years, were served as control group. All the patients and the donators' relatives signed the informed consent to the treatment. METHODS: Changes of collagen fibers and actin in 24 normal spermatic veins and 30 varicocele veins were studied quantitatively and qualitatively by the methods of V-G staining, immunohistochemistry, and scanning electron microscopy. The image analysis was conducted. MAIN OUTCOME MEASURES: Changes of smooth muscle cells (SMC), collagen fibers, type Ⅰ collagen, type Ⅲ collagen and ?-smooth muscle actin, as well as optical density and area density. RESULTS: ①Tunica intima hyperplasia and medial thickening were the main histological appearances of varicocele veins, SMC transformed from the contractile phenotype to the synthetic phenotype.②The optical density and area density of ?-actin, type Ⅰ collagen and type Ⅲ collagen were increased significantly compared with the normal veins (P

Objective: To investigate jugular bulb venous oxyg en partial pressure(PjO2)， hemoglobin saturation (SjO2) and the arterial t o jugular bulb venous oxygen content difference(AjDO2) during anesthesia with desflurane and isoflurane in patients with brain tumor. Methods: Fifty-six patients with brain tumor were randomized into desflur ane or isoflurane for maintaining anesthesia. PjO2, SjO2 and AjDO2 in pati ents were measured during normoventilation, hyperventilation and hypoventilation . Results: During normoventilation， SjO2 and PjO2 in desflu rane group was significantly higer than those in isoflurane group（P<0.05 or P<0.01）, and AjDO2 in desflurane group was significantly lower than that in isoflurane group（P<0.05）.Except that PjO2 in desflurane group was si gnificantly higer than that in isoflurane group during hyperventilation （P< 0.01）, there were no differences in SjO2, PjO2 or AjDO2 between the 2 g roups during hyperventilation or hypoventilation. While anesthesia with desflura ne and isoflurane, there was a positive correlation between PaCO2 and SjO2. Conclusion: At the same anesthetic effect concentration, desflur ane can significantly increase SjO2 and PjO2 in comparison to isoflurane un der normoventilation, suggesting that desflurane may have stronger effect of rel axing cerebral vessel than isoflurane.

Objective To explore the protective effect of uridine 5'-triphosphate (UTP) on cerebral ischemia reperfusion injury in rats.Methods One hundred adult male SD rats were randomly divided into five groups (20 each):sham operation group,saline control group,G_ 10 group,G_ 30 group and G_ 90 group.Ischemia was induced by intraluminal suture embolism of mesocephalic artery (MAO) of the rats in all the groups except sham operation group.UTP solution was delivered through an indwelling tail venous catheter via microinfusion pump 30min after the occlusion of mesocephalic artery at a rate of 5ml/(kg?min),and rats in G_ 10,G_ 30 and G_ 90 group received 10,30 and 90?g/kg UTP,respectively.Rats in sham group and saline control group received normal saline.The total fluid volume was the same,1ml/kg,in all groups.Neurological deficit score (NDS) was determined 24h after reperfusion.Eight SD rats were then randomly selected from each group,and water content of cerebral hemispheres was measured.Two rats from each of sham operation group,saline control group and G_ 90 group were selected for studying the ultrastructure of brain tissue by electron microscopy.Infarct volume was determined by 2,3,5-triphenyl-tetrazolium chloride (TTC) staining with another 10 rats in each group.Results The protective effect of UTP was determined on cerebral ischemia-reperfusion injury in rats,and in G_ 90 group showed the most significant protective effect.For the rats in saline group and G_ 90 group,NDS score was 0.7 and 1.8 (P