ABSTRACT:Objective To explore role of U0126,the specific inhibitor of ERK signaling pathway,in early brain injury (EBI)and the autophagy of nerve cells in hippocampus area in subarachnoid hemorrhage (SAH). Methods A total of 48 male adult SD rats were randomly divided into control group,SAH group,DMSO+SAH group,and U0126+SAH group,with 12 in each.We established SAH rat model by the puncture of internal carotid artery.The same amount of saline water,DMSO and U0126 solution of 0.5 mL per rat was injected respectively into the rats of different groups 30 min before modeling.The rats were killed at 24 h.To measure brain water content by Wet and dry method after 24 h,the morphological changes of hippocampus CA1 neural cells were observed by microscopy;the expression levels of ERK,Beclin-1 and LC3 were detected by using immunohistochemical method. Results Compared with that in sham group,brain water content increased obviously in SAH model group.The density of surviving neurons in SAH group was significantly lower than that in control group (P<0 .0 5 ).ERK signaling pathway was activated obviously,the expressions of Beclin 1 and LC3-Ⅱ were significantly higher than those in control group (P<0.05).Compared with SAH model group,in U0126 group brain water content increased obviously.Compared with those in SAH group,the density of surviving neurons was significantly lower (P<0.05), ERK signaling pathway was suppressed,the expressions of Beclin-1 and LC3-Ⅱ were significantly lower (P<0.05). Conclusion The U0126,the ERK signaling pathway inhibitor,can inhibit neuron autophagy and increase EBR of SAH.

Human immunodeficiency virus (HIV)-1 infection changes transcriptional profiles and regulates. the factors and machinery of the host that facilitate viral replication. Our previous study suggested that the serine/threonine kinase citron kinase (citK) promotes HIV-1 egress. To ascertain if HIV-1 infection affects citK expression in primary cells, peripheral blood mononuclear cells were infected with vesicular stomatitis virus G protein (VSV-G)-pseudotyped HIV-1 vector NL4-3-luc viruses, which resulted in remarkably increased expression of citK. citK overexpression led to a more than two-fold increase in HIV-1 production, whereas a significant decrease was observed when citK was depleted in CD4+ T cells. Infection with HIV-1 pseudoviruses induced increases in the mRNA and protein levels of citK by 2. 5- and 2. 7-fold in HEK293T cells, respectively. By cloning the 5-kb promoter of citK into a luciferase reporter system and transfecting the construct into HEK293T cells, enhanced luciferase activity was observed during HIV-1 infection. Taken together, these data demonstrate that HIV-1 infection upregulates citK expression at the transcriptional level, and thereby renders the host more susceptible to invasion by HIV-1.
CD4-Positive T-Lymphocytes ; virology ; Cloning, Molecular ; Gene Expression Regulation, Enzymologic ; HEK293 Cells ; HIV-1 ; physiology ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Up-Regulation ; Virus Replication

Hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs) is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytosis and the accumulation of internal vesicles in multivesicular bodies. Citron kinase is a Ser/The kinase that we previously reported to enhance human immunodeficiency virus type 1 (HIV-1) virion production. However, the relationship between Hrs and citron kinase in HIV-1 production remains elusive. Here, we report that Hrs interacts with citron kinase via its FYVE domain. Overexpression of Hrs or the FYVE domain resulted in a significant decrease in HIV-1 virion production. Depletion of Hrs by RNA interference in HEK293T cells increased HIV-1 virion production and enhanced the activity of citron kinase. These data suggest that Hrs inhibits HIV-1 production by inhibiting citron kinase-mediated exocytosis.
Down-Regulation ; Endosomal Sorting Complexes Required for Transport ; genetics ; metabolism ; Endosomes ; metabolism ; Exocytosis ; Gene Expression ; Gene Silencing ; drug effects ; HEK293 Cells ; HIV Infections ; genetics ; metabolism ; virology ; HIV-1 ; drug effects ; genetics ; growth & development ; Humans ; Immunoprecipitation ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Microscopy, Fluorescence ; Phosphoproteins ; genetics ; metabolism ; Plasmids ; Protein Binding ; drug effects ; genetics ; Protein Interaction Domains and Motifs ; Protein Structure, Tertiary ; Protein Transport ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; pharmacology ; Transfection ; Virion ; drug effects ; genetics ; growth & development ; Virus Release ; drug effects ; Virus Replication ; drug effects

Objective To explore the effect of extracellular regulated protein kinases (ERK) signaling pathway on early brain injury and autophagy of nerve cell in hippocampus area in rats with subarachnoid hemorrhage (SAH). Methods Forty-eight adult male Sprague-Daw-ley rats were randomly divided into sham group, SAH group, SAH+dimethyl sulfoxide (DMSO) group and SAH+U0126 group, with 12 rats in each group. The SAH model was established with puncture of internal carotid artery. The SAH+U0126 group was injected with U0126 0.05 mg/kg;the sham group and SAH group were injected with normal saline, and the SAH+DMSO group was injected with DMSO 30 min-utes before modeling. They were sacrificed 24 hours after modeling. The brain water content was measured with wet and dry method. The morphology changes of neural cells in hippocampus CA1 were observed by HE staining. The expression of phosphorylation ERK (p-ERK), Beclin-1 and LC3-Ⅱwere detected with immunohistochemical method and Western blotting. Results Compared with the sham group, the brain water content increased (P<0.05), the number of survival neurons decreased (P<0.05), the expression of p-ERK, Beclin-1 and LC3-Ⅱincreased in SAH group (P<0.05). Compared with SAH group, the brain water content increased, the number of survival neurons decreased (P<0.05), the expression of p-ERK, Beclin-1 and LC3-Ⅱ decreased in SAH+U0126 group (P<0.05); and no significant difference was found in SAH+DMSO group (P>0.05). Conclusion The activation of ERK signaling pathway may alleviate early brain injury after SAH by regulation of autophagy.

Objective To explore the effect of traditional Chinese massage on upper limbs spasticity in stroke patients who accepted Botulinum toxin type A (BTX-A) injection. Methods 74 stroke patients with upper limbs spasticity were divided into treatment group (n=36)and control group (n=38). The control group received BTX-A injection, and the treatment group received massage after injection. Both groups received rehabilitation training after injection and were assessed with modified Ashworth scale, Fugl-Meyer assessment and modified Barthel index before and 2 weeks, 4 weeks, 8 weeks and 12 weeks after treatment. Results All the scores of assessments improved in both groups after treatment (P<0.001), and there was no significant difference between them (P>0.05). Conclusion Addition of traditional Chinese massage doesn't work more on spasticity of upper limbs after stroke when accepted BTX-A injection.

Diabetes mellitus is a complex metabolic disease involving multiple organ systems in the body.In recent years,its global incidence rate has increased year by year.In China,the blood glucose control of patients with diabetes mellitus who receive oral hypogly-cemic agents or insulin treatment remains poor.In the early disease stages,exercise is important to control blood glucose levels.Recently,many studies have found that the occurrence of type 2 diabetes mellitus was related to declining levels of irisin,an exercise-related muscle factor.Furthermore,studies have found that irisin improved insulin resistance,promoted the production of pancreatic isletβcells,and affected the body's glucose and lipid metabolism.In addition,its levels were also implicated in the occurrence of various complications,such as diabetic nephropathy and diabetes-related cardiovascular diseases.This article summarizes and analyzes the role of irisin in the occurrence and development of diabetes mellitus and further describes its impact and mechanism on various diabetic complications.

As a metabolic disease,obesity increases the risk of many chronic metabolic diseases;in fact,it is an epidemic in the 21st cen-tury.Exercise is an important means of controlling and treating obesity.Irisin,a myocyte cytokine,was discovered and reported in 2012;it is regulated by exercise and involved in the browning of adipose tissue.A recent discovery is irisin's ability to improve obesity,which is embodied in its effect on glucose and lipid metabolism.Irisin can induce the browning of white fat;promote glucose homeostasis;resist the damage of fat cells caused by inflammation,oxidative stress,and other factors;inhibit insulin resistance;and improve glucose,lipid metabolism,and obesity.This review addresses the role of irisin in improving glucose levels,lipid metabolism,and obesity,as well its role in regulating exercise.Further,the prospect of future research is discussed.

AIM:To investigate the expression of CUE domain-containing 2 (CUEDC2) in hepatocellular car-cinoma ( HCC) and to analyze its clinical prognostic significance .METHODS:Total 186 formalin-fixed paraffin-embed-ded tissues obtained from surgical HCC with detailed clinicopathological and follow -up data were used .The expression of CUEDC2 was detected by immunohistochemistry .The relationships between the expression of CUEDC 2 and clinicopatholog-ical characteristics and prognosis were analyzed .RESULTS: The positive rate of CUEDC 2 in HCC was 85.5% ( 159/186), among which, the low expression was 52.2%(97/186) and the high expression was 47.8%(89/186).CUEDC2 expression was correlated with serum alpha-fetal protein (AFP) level, tumor size, tumor number, tumor differentiation and TNM stage (P<0.05).Kaplan-Meier survival curves showed that the patients with high expression of CUEDC 2 were asso-ciated with significantly shorter overall survival and recurrence-free survival than those with low CUEDC 2 expression ( P<0.05).Multivariate Cox regression analysis revealed 3 independent prognostic factors including CUEDC 2 expression, ser-um AFP and tumor number .CONCLUSION:CUEDC2 was expressed in most HCC tissues , which was relevant to tumor growth, tumor differentiation and prognosis .CUEDC2 could be a novel valuable molecular marker to predict the HCC prog-nosis.

Objective The objective of this study was to investigate the abnormal expression of Notch signaling pathway members in pancreatic cancer and its important effect on the development of pancreatic carcinoma. Methods Affymetrix gene expres-sion microarray was used to screen the differentially expressed members of Notch signal pathway in 10 cases of pancreatic carcinoma and its adjacent tissues,and verified by real-time quantitative PCR and Western blotting. The lentivirus expression vector carrying the siRNA fragment of Jagged 2(JAG2)gene was transfected into the pancreatic cancer primary cells to construct the JAG2 gene re-pression-expressing pancreatic cancer cell line. MTT,flow cytometry and Transwell assays were used to analyze cell proliferation, changes of cell cycle and invasive transfer capabilities. Results A total of 512 differentially expressed genes were detected by Affy-metrix gene expression microarray,including 419 up-regulated genes and 93 down-regulated genes. JAG2( up-regulated expres-sion 8. 20 times),NOTCH1(up-regulated expression 3. 74 times),HES1(up-regulated expression 3. 27 times),and NOTCH2(up-regulated expression 3. 16 times)were differentially expressed in Notch signaling pathway. The results of PCR and Western blotting were consistent with those of gene chip. The growth curves of JAG2 gene repressed pancreatic cancer cells and pancreatic cancer pri-mary cells were drawn by the standard OD490 value of d1-d5 by MTT assay. JAG2 gene repressor expression vector could signifi-cantly inhibit the proliferation of pancreatic cancer cells. The cell cycle analysis showed that the apoptosis and the arrested cell cycle at the S phase were significantly increased in pancreatic cancer cells with JAG2 gene repressor expression. The invasive ability was significantly reduced in JAG2 gene repressor expression pancreatic cancer cells(P<0. 05). Conclusion Some members of the Notch signaling pathway are significantly differentially expressed in pancreatic cancer tissues,and repression of this member can affect the growth,cell cycle,invasion and metastasis of pancreatic cancer cells.

Objective To explore the difference in gut microbiota composition between patients with obstructive sleep apnea(OSA)and healthy individuals and the role of gut microbiota in the pathogenesis of OSA.Methods Thirty-nine patients with OSA admitted to our hospital between May and December,2022 and 20 healthy individuals were enrolled in this study.Stool samples were collected from all the participants for analysis of microbiome composition using 16S rRNA high-throughput sequencing analysis.The alpha diversity,beta diversity,and species difference were determined between the two groups and marker species analysis and metabolic pathway function prediction analysis were performed.Results The species diversity(Shannon and Simpson)indexes,richness(observed species)and evenness(Pielou)of gut microbiota were significantly lower in OSA patients than in the healthy individuals(P<0.05).The OSA patients had also a significantly lowered community diversity(P<0.05)with different gut microbial communities from those of the healthy individuals shown by increased relative abundance of potentially pathogenic bacteria such as Pseudomonas and Monocytogenes(P<0.05).LEfSe analysis showed that the abundance of 23 species of gut microbiota differed significantly between the two groups and the OSA patients had significant increases in the abundance of Pseudomonas,Meganomonas,and Fusobacterium(P<0.05).The differential marker flora affected host homeostasis.Random Forest and ROC curve analyses confirmed that Pseudomonas could be used as important biomarkers for a differential diagnosis.Metabolic pathway function prediction analysis showed that biosynthesis function had the greatest contribution to maintaining gut microbiota homeostasis,and Pseudomonas affected the occurrence and progression of OSA by participating in aromatic bioamine degradation and ketogluconic acid metabolic pathway.Conclusion OSA patients have obvious gut microbiota disturbances,and Pseudomonas may affect the development of OSA by participating in substance metabolism to serve as the potential target gut bacteria for OSA treatment.