Objective To investigate the disease assessment and prognosis value of serum copeptin level in patients with acute cerebral infarction (ACI). Methods One hundred first diagnosed ACI patients were selected as ACI group. According to the National Institutes of Health stroke score (NIHSS), the ACI patients were divided into mild (NIHSS<7 scores), moderate (NIHSS 7-15 scores) and severe (NIHSS>15 scores). Sixty cases of healthy subjects were selected as control group. The serum copeptin level was measured by double antibody sandwich enzyme linked immunosorbent assay method in control group and ACI group (onset within 24 h). The NIHSS, Alberta stroke program early CT score (ASPECTS) and modified Rankin score (mRS) onset within 24 h and 14 d were evaluated in patients with ACI, and the mRS 90 d and 180 d after ACI were evaluated. The neurological impairment was assessed by mRS 180 d after ACI, mRS ≤ 2 scores was good prognosis, ≥ 3 scores was poor prognosis. The correlation was analyzed. Results Among the 100 patients with ACI, mild was in 52 cases, moderate in 34 cases, and severe in 14 cases; good prognosis was in 79 cases and poor prognosis in 21 cases. The serum copeptin levels within 24 h of ACI in mild, moderate and severe patients of ACI group were significantly higher than that in control group:(4.82 ± 1.25), (6.39 ± 2.21) and (9.28 ± 3.82) pmol/L vs. (1.95 ± 0.28) pmol/L. The serum copeptin level within 24 h of ACI in moderate patients was significantly higher than that in mild patients, in severe patients was significantly higher than that in moderate patients, and there were statistical differences (P<0.05). Within 24 h of ACI , the ASPECTS in moderate and severe patients were significantly lower than that in mild patients:(10.02 ± 2.10) and (6.24 ± 3.05) scores vs. (12.16 ± 0.84) scores, in severe patients was significantly lower than that in moderate patients, and there were statistical differences (P<0.05). The NIHSS in moderate and severe patients were significantly higher than that in mild patients:(10.68 ± 3.14) and (16.20 ± 4.26) scores vs. (4.35 ± 1.52) scores, in severe patients was significantly higher than that in moderate patients, and there were statistical differences (P<0.05). The serum copeptin levels within 24 h of ACI and NIHSS in each time point in good prognosis patients were significantly lower than those in poor prognosis patients:(3.52 ± 1.26) pmol/L vs. (8.68 ± 3.06) pmol/L and (5.68 ± 2.11) scores vs. (15.36 ± 3.25) scores, (4.85 ± 1.86) scores vs. (12.60 ± 3.89) scores, (3.68 ± 1.21) scores vs. (6.35 ± 2.96) scores, (2.16 ± 0.75) scores vs. (5.21 ±1.96) scores, and the ASPECTS within 24 h of ACI was significantly higher than that in poor prognosis patients:(11.38 ± 2.21) scores vs. (7.86 ± 2.49) scores, and there were statistical differences (P<0.05). The single factor Logistic regression analysis results showed that the age, ASPECTS, NIHSS and serum copeptin level were the influencing factors of severity of illness in patients with ACI (OR = 1.21, 5.36, 5.61 and 6.62;95%CI 0.99-1.39, 3.34-9.21, 2.86-7.52 and 1.38-12.64;P=0.04, 0.01, 0.01 and 0.00), and the influencing factors of poor prognosis (OR=1.32, 5.21, 4.86 and 6.82;95%CI 0.84-1.43, 3.52-8.39, 2.62-5.35 and 2.67-11.85;P=0.04, 0.01, 0.01 and 0.00). ROC analysis results showed that the area under curve of NIHSS, serum copeptin level and ASPECTS in predicting poor prognosis in patients with ACI were 0.926, 0.863 and 0.624. In the mild, moderate and severe patients, the serum copeptin level was negative correlated with ASPECTS ( r=-0.682,-0.594 and-0.572;P<0.01), and the serum copeptin level was positively correlated with NIHSS ( r = 0.652, 0.614 and 0.586; P<0.01). Conclusions The serum copeptin level in patients with ACI is significantly elevated. The serum copeptin level is positively correlated with neurologic impairment severity and prognosis in patients with ACI, and it has important significance in evaluating pathogenetic condition and prognosis.

AIM:To examine the latent membrane protein 1(LMP1)-DNA sequence in nasopharyngeal carcinoma(NPC) and detect mRNA expression of LMP1, EBNA1, EBNA2, and to explore the relationship between EBV infectious status, expression products and NPC carcinogenesis.METHODS: LMP1 DNA was detected in NPC by PCR. Direct sequence was applied to analyze the difference between NPC-LMP1-DNA and B95-8-LMP1-DNA. mRNA expressions of LMP1, EBNA1, EBNA2 in NPC were detected by nested RT-PCR.RESULTS: LMP1 DNA existed in all 47 NPC tissues. Several single nucleotide variations were found between NPC-LMP1-DNA and B95-8-LMP1-DNA. The notable variation was the lost of XhoⅠrestriction site in NPC. Direct sequence showed 30 bp deletion in NPC. The mRNA expressions of LMP1, EBNA1 and EBNA2 in NPC were 76.6%, 80.0% and 74.5% respectively by nested RT-PCR. The expression of EBNA1 in NPC was promoted by Q promoter while the expression of EBNA1 in B95-8 was promoted by C promoter.CONCLUSION: The way of EBV involved in NPC is complex. Latent genes such as LMP1, EBNA1 and EBNA2 as well as early lytic gene BARF1 may all play certain roles in NPC carcinogenesis.

Objective To investigate the dynamic changes of serum interleukin(IL)-18,IL-1β and copeptin in acute cerebral infarction(ACI),and analyze the relationship with the serious degree of ACI.Methods The levels of serum IL-18,IL-1β and copeptin were measured by Double-antibody sanduicb enzyme-linked immunosorbent assay(ELISA)in 83 patients with ACI who were admitted to the hospital within 24 h,and neurological impairment were evaluated by European stroke scale(ESS)at the 1st,3rd and 7th day respectively after hospitalization.At the same time,the patients with ACI were compared with 32 normal adults.Results The levels of serum IL-18,IL-1β and copeptin at the 1st,3rd and 7th day after hospitalization were(131.30±31.62),(168.30±28.12),(141.26±24.23)ng/L,(0.35±0.04),(0.82±0.10),(0.52±0.21)μ g/L,and(3.64±0.26),(4.18±0.53),(3.26±2.41)μ g/L respectively.There were significant differences among different times respectively(P＜0.05),furthermore they were higher than those in normal adults[(119.12±27.42)ng/L,(0.21±0.08)μ g/L,(2.63±0.23)μ g/L](P＜0.05).Univariate Logistic regression analysis showed that age,copeptin,IL-18,IL-1β and ESS score was the influencing factor in the serious degree of ACI.There was no significant difference in assessing the serious degree of ACI by receiver operating characteristic(ROC)curve between IL-18 and age,copeptin,IL-1 β,ESS score(P＞0.05).Conclusions The serum levels of IL-18,IL-1β and copeptin are increasing after ACI,indicating that the inflammatory and immune factors may be involved in the development process of ACI.The serum levels of IL-18,IL-1β and copeptin can reflect the serious degree of ACI.

AIM: Our study focused on investigation of tissue specific regulatory activity of the newly cloned promoter: PLUNC-p with driving enhanced green fluorescent protein ( EGFP ). METHODS: Transgenic Xenopus Laevis system was applied.RESULTS: The green fluorescence protein directed under PLUNC-p was expressed strictly in branchial arches and epidermis of Xenopus Laevis embryos while CMV promoter showed ubiquitous regulation characteristic.CONCLUSION: PLUNC-p is able to direct epithelia specific expression of EGFP . This property of PLUNC-p might raise the possibility that lead target genes to express tissue-specifically.

Malignant tumors continue to pose a significant threat to human life and safety and their development is primarily due to the activation of proto-oncogenes and the inactivation of suppressor genes.Among these,the activation of proto-oncogenes possesses greater potential to drive the malignant transformation of cells.Targeting oncogenes involved in the malignant transformation of tumor cells has provided a novel approach for the development of current antitumor drugs.Several preclinical and clinical studies have revealed that the development pathway of B cells,and the malignant transformation of mature B cells into tumors have been regulated by oncogenes and their metabolites.Therefore,summarizing the key oncogenes involved in the process of malignant transformation of mature B cells and elucidating the mechanisms of action in tumor development hold significant importance for the clinical treatment of malignant tumors.

Objective: To understand the influence of experiential health education on diet control of college students with pre diabetes mellitus, and to provide reference for healthy eating habits promotion among college students. Methods: According to the method of random number table, 78 pre diabetic college students screened from Changzhi Medical College from September 2020 to June 2021 were randomly assigned to observation group and control group (39 students in each group). The control group received routine health education for 10 months, once a week for 1 hour each time; On the basis of the above, the observation group received experiential health education once a week for 1 hour, including diet experience, exercise experience, blood sugar test experience and chronic complications experience. Blood glucose and lipids level, body mass index (BMI), dietary control as well as the stages of change for dietary control behavior were compared between the two groups. Results: There was significant difference between the observation group and the control group in the stages of change for dietary control behavior 10 months after intervention ( χ 2=8.92, P <0.05). The compliance score of the observation group was significantly higher than that of the control group in the same period 10 months after the intervention ( t =3.74, P <0.01), the score of the knowledge of diet control in the observation group 10 months after intervention was significantly higher than that in the control group ( t =11.51, P <0.05). The levels of BMI, TG and TC in the observation group were significantly lower than those in the control group at 5 and 10 months after intervention, and the differences were statistically significant ( P <0.05). Conclusion Experiential health education helps to promote awareness of diabete related knowledge, enhance self management behavior and good diet control habits, and is effective for blood glucose control.

Objective To study the distributions of three single nucleotide polymorphisms (SNPs) in β-arrestin2 (ARRB2) which including rs3786047,rs1045280 and rs2036657 and to elucidate the relationship between these SNPs and response to methadone maintenance treatment (MMT) among heroin-dependent patients of Han ethnicity population in Hunan.Methods Han MMT patients were recruited in four random-chosen MMT clinics from Hunan province.Demographics,history of drug-use and MMT were recorded.ARRB2 SNPs were genotyped to determine the association between SNPs and response to MMT.Results Distributions of the three SNPs were in Hardy-Weinberg equilibrium in both groups (responders vs.non-responders).There was no statistical significance in the distribution frequency of genotype on rs3786047 (x2=0.486 2,P=0.784),rs1045280 (x2=1.591 9,P=0.451) and rs2036657 (x2=1.061 5,P=0.588) in ARRB2 among the responders or the non-responders.Conclusion Associations between the ARRB2 genotypes,rs3786047,rs1045280 and rs2036657,and MMT response in Han MMT patients in Hunan province did not appear.

OBJECTIVE To study the intervention effects and mechanism of Compound yu ’e nasal drops on ovalbumin induced allergic rhinitis in rats . METHODS The allergic rhinitis model of rat was induced with ovalbumin . Model rats were randomly divided into model group ，triamcinolone acetonide group （positive control ，0.026 mg/kg），Compound yu ’e nasal drops high-dose，medium-dose and low -dose groups （134.4、67.2、33.6 mg/kg），12 rats in each group . Another blank control group was set. Except for blank control group ，the corresponding drugs were given by nasal drip twice a day for 14 days. One hour after last administration，the nasal symptom scores of rats were recorded ；the levels of serum immunoglobulin E （IgE），interleukin-2（IL- 2），IL-13 and tumor necrosis factor -α（TNF-α）were measured by enzyme -linked immunosorbent assay . The changes of nasal mucosa in rat were observed by HE staining . The expressions of TNF -α，IL-2 and IL -13 in nasal mucosa were detected by Western blot. RESULTS Compared with blank control group ，nasal symptom score and the levels of serum IgE ，IL-2，IL-13，TNF-α in model group were increased significantly （P＜0.01）；obvious pathological injury was found in nasal mucosa ，and the expressions of TNF -α，IL-2 and IL -13 protein were increased significantly （P＜0.01）. Compared with model group ，Compound yu ’e nasal drops significantly reduced the nasal symptom score ，the levels of serum IgE ，IL-2，IL-13，TNF-α to different extents ，improved pathological injury of nasal mucosa and significantly inhibited the expressions of TNF -α，IL-2 and IL -13 protein（P＜0.05 or P＜ 0.01）. CONCLUSIONS Compound yu ’e nasal drops play significant effects against allergic rhinitis in rats by regulating the balance of t ype 1 helper T cells/type 2 helper T cells ，balancing and inhibiting the secretion of inflammatory cytokines .

OBJECTIVES: Liver cancer is the sixth most common malignant tumor in the world. Hepatocellular carcinoma (HCC) accounts for 85%-90% of all patients with liver cancer. It possesses the characteristics of insidious onset, rapid progression, early recurrence, easy drug resistance, and poor prognosis. NIMA related kinase 2 (NEK2) is a cell cycle regulating kinases, which regulates cell cycle in mitosis. Cellular senescence is a complex heterogeneous process, and is a stable form of cell cycle arrest that limits the proliferative potential of cells. This study aims to investigate the relationship between the expression level of NEK2 and the senescence in hepatoma cells, and to explore the effect of NEK2 expression on hepatoma cell senescence and the underlying molecular mechanism. METHODS: A total of 581 senescence-relevant genes were obtained from the GenAge website. The gene expression data of tumor tissues of 370 HCC patients were downloaded from the Cancer Genome Atlas database. The co-expression of NEK2 and aging-related genes was analyzed by R-package. KEGG was used to analyze the significant gene enrichment pathway of differentially expressed genes in NEK2 overexpression HEK293. The stable transfected cell lines with overexpression and knockdown of NEK2 were constructed in hepatoma cell line SMMC-7721 and HepG2, and senescence-associated β-galactosidase (SA-β-gal) staining was used to detect senescence, the cell proliferation was detected by CCK-8 method and clone formation experiment, the cell cycle was analyzed by flow cytometry, and the expression of proteins related to p53/p21, p16/Rb, and phosphatase and tensin homolog deleted on chromosome ten (PTEN)/Akt signal transduction pathway was detected by Western blotting. RESULTS: There were 320 senescence related genes co-expressed with NEK2. KEGG analysis showed that the senescence signaling pathway was significantly enriched in HEK293 cells with overexpression of NEK2.Compared with SMMC-7721 or HepG2 without knockdown of NEK2, the senescent cells of SMMC-7721 and HepG2 with knockdown of NEK2 were increased, cell proliferation and clone formation were decreased significantly, the percentage of cells in G₀/G₁ phase was increased, the expression levels of phospho-Akt (p-Akt) and phospho-Rb (p-Rb) protein were decreased significantly, and the expression level of p16 protein was increased significantly (all P<0.05). Compared with SMMC-7721 or HepG2 transfected with blank plasmid, the senescent cells of SMMC-7721 and HepG2 overexpressing NEK2 were decreased, the cell proliferation and clone formation were increased significantly, the percentage of cells in G₀/G₁ phase were decreased, the expression levels of p-Akt and p-Rb protein were increased significantly, and the expression level of p16 protein was decreased significantly (all P<0.05). CONCLUSIONS NEK2 may mediate the anti-aging effect of hepatoma cells through p16/Rb and PTEN/Akt signal transduction pathways, which provides a new theoretical basis for NEK2 to promote the progress of liver cancer and a new idea for the targeting treatment for liver cancer.
Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Proliferation/physiology* ; Cellular Senescence/genetics* ; HEK293 Cells ; Humans ; Liver Neoplasms/pathology* ; NIMA-Related Kinases/genetics* ; Proto-Oncogene Proteins c-akt/metabolism*

OBJECTIVES: Multiple myeloma (MM) is a plasma cell malignancy occurring in middle and old age. MM is still an incurable disease due to its frequent recurrence and drug resistance. However, its pathogenesis is still unclear. Abnormal amino acid metabolism is one of the important characteristics of MM, and the important metabolic pathway of amino acids participates in protein synthesis as basic raw materials. Aminoacyl transfer ribonucleic acid synthetase (ARS) gene is a key regulatory gene in protein synthesis. This study aims to explore the molecular mechanism for ARS, a key factor of amino acid metabolism, in regulating amino acid metabolism in MM and affecting MM growth. METHODS: The corresponding gene number was combined with the gene expression profile GSE5900 dataset and GSE2658 dataset in Gene Expression Omnibus (GEO) database to standardize the gene expression data of ARS. GSEA_4.2.0 software was used to analyze the difference of gene enrichment between healthy donors (HD) and MM patients in GEO database. GraphPad Prism 7 was used to draw heat maps and perform data analysis. Kaplan-Meier and Cox regression model were used to analyze the expression of ARS gene and the prognosis of MM patients, respectively. Bone marrow samples from 7 newly diagnosed MM patients were collected, CD138⁺ and CD138^- cells were obtained by using CD138 antibody magnetic beads, and the expression of ARS in MM clinical samples was analyzed by real-time RT-PCR. Human B lymphocyte GM12878 cells and human MM cell lines ARP1, NCI-H929, OCI-MY5, U266, RPMI 8266, OPM-2, JJN-3, KMS11, MM1.s cells were selected as the study objects. The expression of ARS in MM cell lines was analyzed by real-time RT-PCR and Western blotting. Short hairpin RNA (shRNA) lentiviruses were used to construct gene knock-out plasmids (VARS-sh group). No-load plasmids (scramble group) and gene knock-out plasmids (VARS-sh group) were transfected into HEK 293T cells with for virus packaging, respectively. Stable expression cell lines were established by infecting ARP1 and OCI-MY5 cells, and the effects of knockout valyl-tRNA synthetase (VARS) gene on proliferation and apoptosis of MM cells were detected by cell counting and flow cytometry, respectively. GEO data were divided into a high expression group and a low expression group according to the expression of VARS. Bioinformatics analysis was performed to explore the downstream pathways affected by VARS. Gas chromatography time-of-flight mass spectrometry (GC-TOF/MS) and high performance liquid chromatography (HPLC) were used to detect the valine content in CD138⁺ cells and ARP1, OCI-MY5 cells and supernatant of knockdown VARS gene in bone marrow samples from patients, respectively. RESULTS: Gene enrichment analysis showed that tRNA processing related genes were significantly enriched in MM compared with HD (P<0.0001). Further screening of tRNA processing-pathway related subsets revealed that cytoplasmic aminoacyl tRNA synthetase family genes were significantly enriched in MM (P<0.0001). The results of gene expression heat map showed that the ARS family genes except alanyl-tRNA synthetase (AARS), arginyl-tRNA synthetase (RARS), seryl-tRNA synthetase (SARS) in GEO data were highly expressed in MM (all P<0.01). With the development of monoclonal gammopathy of undetermined significance (MGUS) to MM, the gene expression level was increased gradually. Kaplan-Meier univariate analysis of survival results showed that there were significant differences in the prognosis of MM patients in methionyl-tRNA synthetase (MARS), asparaginyl-tRNA synthetase (NARS) and VARS between the high expression group and the low expression group (all P<0.05). Cox regression model multivariate analysis showed that the high expression of VARS was associated with abnormal overall survival time of MM (HR=1.83, 95% CI 1.10 to 3.06, P=0.021). The high expression of NARS (HR=0.90, 95% CI 0.34 to 2.38) and MARS (HR=1.59, 95% CI 0.73 to 3.50) had no effect on the overall survival time of MM patients (both P>0.05). Real-time RT-PCR and Western blotting showed that VARS, MARS and NARS were highly expressed in CD138⁺ MM cells and MM cell lines of clinical patients (all P<0.05). Cell counting and flow cytometry results showed that the proliferation of MM cells by knockout VARS was significantly inhibited (P<0.01), the proportion of apoptosis was significantly increased (P<0.05). Bioinformatics analysis showed that in addition to several pathways including the cell cycle regulated by VARS, the valine, leucine and isoleucine catabolic pathways were upregulated. Non-targeted metabolomics data showed reduced valine content in CD138⁺ tumor cells in MM patients compared to HD (P<0.05). HPLC results showed that compared with the scramble group, the intracellular and medium supernatant content of ARP1 cells and the medium supernatant of OCI-MY5 in the VARS-shRNA group was increased (all P<0.05). CONCLUSIONS MM patients with abnormal high expression of VARS have a poor prognosis. VARS promotes the malignant growth of MM cells by affecting the regulation of valine metabolism.
Humans ; Valine-tRNA Ligase ; Multiple Myeloma/genetics* ; Metabolomics ; Amino Acids ; RNA, Transfer