Objective To observe the Leu72Met single nucleotide polymorphism (SNP)of ghrelin gene and the relationship with essential hypertension (EH).Methods Polymerase chain reaction -restriction fragment length polymorphism (PCR -RFLP)was used to detect the Leu72Met SNP of ghrelin gene in 210 EH patients and 220 healthy controls.The plasma ghrelin was detected by radioimmunoassay method collected from all subjects. Results There were three types of polymorphism of ghrelin gene at the base site Leu72Met.There were significant differences in the genotypes (CC,CA,AA)and alleles (C,A)between the EH patients and the controls (χ2 =6.054,P =0.048;χ2 =5.866,P =0.015).In EH group,the plasma ghrelin level in subjects who were homozygous CC without mutant was not only significantly lower than those who were heterozygous CA,but also lower than those who were nucleotide homozygous mutant AA (t =-8.738,P =0.000;t =-5.103,P =0.000).The patients with CC genotype had higher SBP (t =4.298,P =0.000;t =2.236,P =0.019)and lower HDL -C (t =-11.682,P =0.000;t =-7.872,P =0.000).The patients with A allele had lower plasma ghrelin (t =-16.264,P =0.000), HDL -C (t =-15.332,P =0.000)and higher SBP(t =3.800,P =0.000),DBP(t =11.895,P =0.000),and LDL -C (t =38.401,P =0.000).Conclusion The Leu72Met SNP of ghrelin gene is significantly related to the susceptibility of EH.Base mutation C to A reduced the incidence of EH.The Leu72Met polymorphism of ghrelin gene is related to the plasma ghrelin,blood pressure and blood lipid metabolism.Base mutation C to A elevated plasma ghrelin,and lowered blood pressure and blood lipid.

Objective To analyze the prevalence status and the genetic characterizations of influenza B viruses isolated in Hunan Province after pandemic influenza A (H1N1) 2009,and to explore possible reasons for the prevalence.MethodsThroat swabs were collected from outpatients with influenza-like illness in 23 sentinel hospitals of Hunan Province in 2010.Influenza viruses were isolated with Madin-Darby canine kidney (MDCK) cells and identified by haemagglutination inhibition test.The genomes of 10 selected influenza B viruses were sequenced and analyzed for phylogenetic and molecular characterization.ResultsWith the reduction of isolation of pandemic influenza A (H1N1)2009 viruses,influenza B virus became the predominant isolated strain in the first half of 2010.Epidemic viruses mainly belonged to the B/Victoria lineage,and both two lineages co-circulated.Seven out of 11 influenza outbreaks caused by type B.Ten strains were filled into 2 branches of BV and BY which were classified by their lineage types in polymerase (PB2,PB1,PA),hemagglutinin (HA),neuraminidase (NA),NB,membrane protein (M1),influenza B virus membrane protein M2 (BM2),and non-structural protein (NS1,NS2) phylogenetic trees except the NP phylogenetic tree in which 10 strains were all in the BY branch.Compared with World Health Organization (WHO) vaccine strains,the amino acid identity of 11 proteins of the 10 strains was high (97.2％-100.0％).However,some amino acid point mutations were found.No mutation was found in drug resistance mutation sites.Some mutations in NA,NB,PB1,PB2 and NS2 molecules were found in 2 strains isolated from outbreaks compared with strains from sentinel surveillance.Conclusions The point mutations,insertions and genetic reassortment indicate viruses sustaining evolution,which is probably the reason for predominant influenza B viruses after pandemic influenza A (H1N1) 2009 in Hunan Province.

Objective:To screen and identify autoantibody biomarker to diagnose patients with nasopharyngeal carcinoma (NPC).Methods:Candidate autoantibodies against tumor-associated antigens were identified from NPC CNE2 cells using serological proteome analysis. Levels of candidate autoantibody biomarkers were measured by enzyme-linked immunosorbent assay (ELISA) in 50 patients with NPC and 80 normal controls recruited from the Cancer Hospital of Shantou University Medical College between July 2014 and January 2015. Receiver operating characteristic curve (ROC) was employed to evaluate diagnostic efficacy.Results:Serological proteome analysis showed that sera from patients with NPC yielded a positive spot, of which was identified as enolase 1 (ENO1). ELISA results showed that the level of serum autoantibody against ENO1 in patients with NPC was significantly higher than that in normal controls [0.165 (0.088, 0.378) vs. 0.100 (0.054, 0.117), Z=4.077, P<0.001]. With the optimum diagnostic cutoff of 0.164, ROC curve showed the diagnostic sensitivity and specificity of autoantibodies against ENO1 were 52.0% and 90.0%, respectively. Measurement of autoantibody against ENO1 demonstrated a positive rate of 75.0% for early stage NPC. Conclusion:Autoantibody against ENO1 may be a potential diagnostic biomarker for NPC.

Objective:To systematically review the effect of preoperative pulmonary rehabilitation on postoperative pulmonary complications in patients with lung cancer.Methods:The randomized controlled trials of preoperative pulmonary rehabilitation for lung cancer published in the past 10 years were retrieved in China Biomedical Literature Database, China National Knowledge Infrastructure, VIP, WanFang Data, PubMed, Embase, Web of Science, Cochrane Library and Wiley Online Library. The search limit was from January 1, 2010 to January 1, 2021. The quality evaluation and data extraction of the included article were carried out, and the Meta-analysis of the outcome indicators of postoperative pulmonary complications was conducted.Results:A total of 12 articles were included. Meta-analysis showed that the incidence of postoperative pulmonary complications in the intervention group was lower than that in the control group, and the difference was statistically significant [ RR=0.42, 95% CI (0.32, 0.55) , P<0.01]. Subgroup analysis was performed according to the duration of intervention, and the results showed that when the intervention duration was one week, the difference between the intervention group and the control group was statistically significant [ RR=0.36, 95% CI (0.24, 0.55) , P<0.01], when the intervention duration was more than one week, the difference between the intervention group and the control group was statistically significant [ RR=0.48, 95% CI (0.33, 0.68) , P<0.01) ]. Subgroup analysis was conducted with different evaluation time after operation, and the results showed that when the evaluation time was one month, the difference between the intervention group and the control group was statistically significant [ RR=0.48, 95% CI (0.34, 0.68) , P<0.01], when the evaluation time was more than one month, there was no significant difference between the intervention group and the control group [ RR=0.39, 95% CI (0.08, 1.85) , P=0.24]. Except for pneumonia or pulmonary infection, atelectasis, and bronchopleural fistula, there was no significant difference in the incidence of postoperative pulmonary complications in patients with lung cancer ( P>0.05) . Conclusions:Exercise that includes breathing training for at least one week is a common intervention for preoperative pulmonary rehabilitation. Preoperative pulmonary rehabilitation is beneficial to reduce the overall and one month postoperative pulmonary complications after lung cancer surgery, but the impact on postoperative pulmonary complications evaluated for more than one month and specific complications requires further study.

IL-34 can promote osteoclast differentiation and activation, which may contribute to steroidinduced osteonecrosis of the femoral head (ONFH). Animal model was constructed in both BALB/c and IL-34 deficient mice to detect the relative expression of inflammation cytokines. Micro-CT was utilized to reveal the internal structure. In vitro differentiated osteoclast was induced by culturing bone marrow-derived macrophages with IL-34 conditioned medium or M-CSF. The relative expression of pro-inflammation cytokines, osteoclast marker genes, and relevant pathways molecules was detected with quantitative real-time RT-PCR, ELISA, and Western blot. Up-regulated IL-34 expression could be detected in the serum of ONFH patients and femoral heads of ONFH mice. IL-34 deficient mice showed the resistance to ONFH induction with the up-regulated trabecular number, trabecular thickness, bone value fraction, and down-regulated trabecular separation. On the other hand, inflammatory cytokines, such as TNF-α, IFN-γ, IL-6, IL-12, IL-2, and IL-17A, showed diminished expression in IL-34 deficient ONFH induced mice. IL-34 alone or works in coordination with M-CSF to promote osteoclastogenesis and activate ERK, STAT3, and non-canonical NF-κB pathways. These data demonstrate that IL-34 can promote the differentiation of osteoclast through ERK, STAT3, and non-canonical NF-κB pathways to aggravate steroid-induced ONFH, and IL-34 can be considered as a treatment target.

Cell metabolomics is an important branch of metabolomics, which could dynamically monitor cell response and metabolic changes after drugs acting on cells, and look for potential biomarkers. Cell metabolomics has been widely used in illustration of disease mechanism, evaluation of drug efficacy and development of new drug through elucidating the pathophysiological mechanism of the disease and the effect of drug treatment intervention. The researches process of cellular metabolomics and its application in central nervous system diseases were reviewed in order to provide theoretical basis for in-depth study of the pathogenesis and prevention and treatment of central nervous system diseases.

Objective To construct a cardiovascular chip model for evaluating the damage of vascular glycocalyx induced by four marine toxins: okadaic acid (OA), conotoxin (CTX), tetrodotoxin (TTX) and gymnodimine (GYM), and explore the protective effect of triptolide on toxin-induced injury. Methods Human umbilical vein endothelial cells（HUVEC) were inoculated into a three-channel microfluidic chip. CCK-8 method and immunofluorescence staining were used to analyze the damage of cell viability and glycocalyx tissue induced by low, middle and high concentrations of marine toxin, as well as the protective effect of triptolide on toxin-induced injury. Results The cells in the cardiovascular chip grew well and had structurally intact glycocalyx. Compared with the control group, the activity of HUVEC cells were inhibited in group of the medium and high concentration of OA and high concentration of GYM (P<0.05). The activity of cells had not been inhibited by CTX and TTX significantly , but all the four toxins caused serious damage to the glycocalyx tissue (P<0.01). After pre-protection with triptolide, the toxicity of the four toxins to HUVEC cells and the damage rate of glycocalyx decreased significantly. Conclusion The four marine biotoxins could damage the activity and glycocalyx of HUVEC cells in a dose-dependent manner, while triptolide has a protective effect on HUVEC cells injured by toxin.

Objective: To analyze the genome characteristics of an avian influenza A (H9N2) virus isolated from an 11-month-old infant, and to look for possible sources of infection. Methods: Throat swabs were collected from an infant with influenza-like illness in influenza sentinel surveillance hospitals and isolated for influenza viruses using cells. The isolates were identified for influenza virus types and subtypes by the method of hemagglutination assay, hemagglutination inhibition assay and fluorescence PCR. Whole genome sequencing of the isolated virus was carried out. The genome nucleic acid sequences and the deduced amino acid sequences were analyzed by comparing the phylogenetic trees which were constructed by bioinformatics software. Results: A seasonal un-typed influenza virus was isolated from the infant with influenza like illness. With fluorescent PCR method , it was identified as H9N2 subtype of avian influenza virus and the case was confirmed as a human infected with an avian influenza A(H9N2) virus. Epidemiological studies revealed that the case had no clear history of poultry contact and exposure. Blast analysis shows that eight segments of the viral genome are avian origin, and 97.5%-99.8% homology with that of viruses isolated from the live-poultry markets. The virus belongs to G57 genotype, deduced amino acid sequence analysis shows that the virus has typical low pathogenic avian influenza characteristics. Conclusions Although the case does not have a clear history of contact or exposure to poultry, molecular traceability suggests that possible sources of infection may be still from poultry.

Objective To identify the potential long non-coding RNA (lncRNA) expressed in rheumatoid arthritis (RA) synovium key to RA onset and investigate its association with immune cell infiltration. Methods RA synovium data were downloaded from the GEO database and normalized. The lncRNAs key to RA onset were identified using multiple machine learning methods. Infiltration of 22 immune cell populations in RA synovium was measured by cell-type identification by estimating relative subsets of RNA transcripts (CIBER-SORT). The relationship between the key lncRNA and infiltrating immune cells was analyzed. Finally, real-time quantitative PCR was applied to validate the expression of the key lncRNA in RA synovial cells. Results lncRNA human leukocyte antigen complex P5(HCP5) was identified as the key lncRNA associated with RA onset. Infiltration analysis revealed increased abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages while decreased abundance of M2 macrophages in RA synovial tissue. Correlation analysis demonstrated that the lncRNA HCP5 expression was positively associated with the infiltration abundance of CD8⁺ T cells, γδ T cells, and M1 macrophages in RA synovial tissue. Furthermore,the expression of lncRNA HCP5 in RA synovial cells was up-regulated. Conclusion lncRNA HCP5 expression is up-regulated in RA synovial tissue and potentially associated with immune cells infiltration.
Humans ; Arthritis, Rheumatoid ; CD8-Positive T-Lymphocytes ; HLA Antigens/metabolism* ; RNA, Long Noncoding/metabolism* ; Synovial Membrane/metabolism*

Objective To construct a glioma microfluidic chip model to simulate tumor microenvironment for evaluating the medicinal efficacy of anti-glioma traditional Chinese medicines. Methods Glioblastoma cells U251 were seeded into microfluidic chips with different culture modes, and the cell viability and tumour microenvironment within the constructed model were characterized. Fluorescence staining was used to evaluate the effects of the positive drugs temozolomide （TMZ） and docetaxel （DOC） on the cell activity and apoptosis within the model, which was applied to evaluate the medicinal efficacy of the extracts of the herb Scutellaria barbata on gliomas. Results The cells in the constructed U251 microfluidic chip model displayed high viability and were able to mimic the hypoxic microenvironment of tumor to a certain extent. The viability of the U251 cells in the microfluidic chips decreased with the increasing of the concentration of the positive drug, and the viability of the 3D cultured U251 cells was higher than that in the 2D condition （P<0.05）. The intracellular mitochondrial membrane potential decreased with the increasing of the concentration of the positive drug. And the 2 mg/ml Scutellaria barbata extract killed U251 cells to a certain extent and reduced the mitochondrial membrane potential of the cells in the model. Conclusion This study successfully constructed a microfluidic chip model of glioma that could effectively simulate the tumor microenvironment and rapidly evaluate the anti-tumor medicinal efficacy, which provided a new strategy for the medicinal efficacy evaluation and active components screening of anti-glioma traditional Chinese medicines.