Objective To investigate the expression of miR-6768-5p in lung cancer tissue and its effect on the proliferation and invasion of lung cancer cells through targeted regulation of carboxypeptidase A4(CPA4).Methods The expression of miR-6768-5p in lung cancer tissues and adjacent tissues was analyzed u-sing the TCGA database.Quantitative real-time PCR(qPCR)was used to detect the expression of miR-6768-5p in human lung cancer cell lines(HCC1588,H1650,H1299,A549,HCC827)and normal alveolar epithelial cells(HPAEpiC cells).Lung cancer cells were transfected with NC mimics and miR-6768-5p mimics,respec-tively,and divided into NC group and miR-6768-5p group.The MTS assay and Matrigel invasion assay were used to detect the cell proliferation and invasion ability of each group,respectively.The putative binding sites of miR-6768-5p and CPA4 were verified using RNAhybrid software and dual-luciferase reporter gene experi-ment.The expression of CPA4 mRNA in each group of cells was detected by qPCR.The expression of AKT/c-MYC signaling pathway proteins in the cells of each group was analyzed by Western blot.Results Com-pared with the adjacent tissues,the relative expression level of miR-6768-5p in lung cancer tissues was signifi-cantly decreased,and the difference was statistically significant(P＜0.05).Compared with HPAEpiC cells,the relative expression level of miR-6768-5p was significantly decreased in lung cancer cell lines,and the differ-ence was statistically significant(P＜0.05).Compared with the NC group,the cell proliferation rate of miR-6768-5p group was significantly decreased(P＜0.05).The number of invasive cells in NC group and miR-6768-5p group was(131.30±12.55)and(37.45±7.77),respectively,and the number of invasive cells in miR-6768-5p group was significantly lower than that in NC group(P＜0.05).The relative expression level of CPA4 mRNA in H1299 cells of miR-6768-5p group was significantly lower than that in NC group(t=4.93,P＜0.05).Compared with the NC group,the expressions of AKT/c-myC signaling pathway proteins p-AKT,p-mTORC1,XIAP,MDM2 and C-myC proteins in miR-6768-5p group were significantly decreased.Conclusion The expression of miR-6768-5p is decreased in lung cancer tissues,and miR-6768-5p may inhibit the activation of AKT/c-MYC signaling pathway by targeting CPA4,and reduce the proliferation and invasion ability of lung cancer H1299 cells.

Objective:To study the biological traits and mutations of the influenza A (H3N2) virus in order to produce a vaccine and offer references for controlling and preventing influenza epidemics.Methods:Four strains of the influenza A(H3N2) virus were chosen from the Huai′an surveillance network laboratory. Nucleic acid extraction, library building, and sequencing (CridION x5 MKI Nanopore) were used to produce the whole-genome sequences. Using homologous alignments of whole-genome sequences, phylogenetic tree construction, and amino acid variant screening, bioinformatics analysis was carried out.Results:The nucleotide identity between 8 gene segments ranged from 97.1% to 100.0%. The gene that differed the most from the reference sequences was HA (97.1%-99.9%), and the gene that differed the least was MP (98.6%-99.9%). The HA gene (3.06%) and MP gene (1.43%) were the regions with the greatest and lowest frequencies of nucleotide site change, respectively. The rates of nucleotide change varied significantly between the genes ( χ2=14.293, P=0.046). Four influenza A(H3N2) virus strains′ whole-genome phylogenies from each of the eight gene segments maintained a roughly consistent topological structure. One strain was linked to the 3C.2a1b.1b clade, which was lost at the 142NWT, 149NGT(HA1), and 436NLS(NA). Three strains were linked to the 3C.2a1b.2a.1a clade lineage. Amantadine and NA inhibitors were effective against all Huai′an strains. Conclusions:The antigenicity of one strain of Huai'an strain changed and its matching with the vaccine strain of that year was low. It is suggested that the genetic surveillance of H3N2 influenza virus should be continuously strengthened to provide scientific basis for influenza prevention and control and influenza vaccine screening.

Objective: To determine the current prevalence of health risk behaviors among college students in Henan Province, and to conduct an in depth analysis of the impact of cumulative ecological risks on health risk behaviors, so as to provide scientific basis for promoting healthy development of adolescents. Methods: Using a multi stage stratified cluster sampling method, 9 743 college students from six universities in Henan Province were included as the research subjects from April to June 2023. A questionnaire survey was conducted using the College Student Cumulative Ecological Risk Scale and the China Urban Adolescent Health Related Behavior Survey Questionnaire (University Version). Data were analyzed by descriptive statistical analysis, Chi square test and binary Logistic regression. Results: The reporting rates of unhealthy eating behavior, unhealthy weight loss behaviors, lack of physical activity, daily risk behaviors, negative emotions, current smoking behavior current drinking behaviors, Internet addiction emotions and dangerous sexual behaviors among college students in Henan Province were 40.2%, 39.5%, 76.0%, 13.7%, 28.1%, 11.3%, 12.7%, 5.9% and 2.2%, respectively. The reporting rates of negative emotions, current smoking behaviors, current drinking behaviors, dangerous sexual behaviors and daily risk behaviors of college students were higher in boys than in girls ( χ 2=44.00, 995.20, 902.49, 121.95, 103.09, P <0.05). In terms of reporting rates of unhealthy diet, unhealthy weight loss and lack of exercise behavior, girls were higher than boys ( χ 2=107.59, 13.01, 145.83, P <0.05). Cumulative ecological risk was positively correlated with overall health risk behaviors. For every unit increase in the cumulative ecological risk index, the risk of health risk behaviors among college students increased by 48%. Conclusions The prevalence of health risk behaviors among college students is relatively common. It should adrocate for a healthy lifestyle, reduce the cumulative ecological risk and the occurrence of health risk behaviors to promote the healthy development of adolescents.

Objective:To investigate the effect of lncRNA AC132217.4 on the proliferation and invasion of liver cancer MHCC97-H cells and its molecular mechanism.Methods:The TCGA database was used to analyze the differential expression of AC132217.4 in liver cancer tissue and adjacent tissue, and to analyze the relationship between the expression level of AC132217.4 and the overall survival of liver cancer patients. Transfection of pcDNA-AC132217.4 plasmid into MHCC97-H cells was defined as AC132217.4 group, transfection of pcDNA plasmid into MHCC97-H cells was defined as negative control (NC) group, respectively. The proliferation and invasion ability of MHCC97-H cells were detected by MTT method and Matrigel invasion assay. The binding site between AC132217.4 and miR-18a-5p was analyzed by starBase v2.0 software and dual luciferase reporter gene assay. Real-time quantitative PCR (RT-qPCR) detected the differential expression of miR-18a-5p in the two groups of MHCC97-H cells. The expression of epithelial-mesenchymal transition protein was detected by Western-blotting. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between the two groups. Results:Compared with adjacent tissues, the expression of AC132217.4 was down-regulated in liver cancer tissues ( P<0.01). Compared with liver cancer patients with low expression of AC132217.4, the overall survival of liver cancer patients with high AC132217.4 expression was longer ( P<0.05). The pcDNA-AC132217.4 plasmid significantly inhibited the proliferation of MHCC97-H cells ( P<0.05). The number of invasive cells in the NC group and AC132217.4 group were (131.30±12.55) and (37.45±7.77), respectively. The pcDNA-AC132217.4 plasmid significantly inhibited the invasive ability of MHCC97-H cells ( t=6.36, P<0.01). AC132217.4 directly complemented miR-18a-5p ( P<0.01). The expression of miR-18a-5p in MHCC97-H cells in AC132217.4 group (1.04±0.30) was significantly lower than that in NC group (6.13±0.75) ( t=6.27, P<0.01). Compared with the NC group, the expressions of epithelial phenotype proteins Cytokeratin and Claudin-1 in MHCC97-H cells in AC132217.4 group were up-regulated, while the expressions of mesenchymal phenotype proteins Vimentin, Slug and Snail were down-regulated. Conclusions:The expression of AC132217.4 is low in liver cancer tissue, and it is related to the overall survival of liver cancer patients. AC132217.4 might inhibit the proliferation and invasion of liver cancer MHCC97-H cells by sponge miR-18a-5p.

Objective To analyze the chemical compounds of Shenqi Dihuang decoction by the ultraperformance liquid chromatography coupled with linear quadrupole ion trap-orbitrap mass spectrometry (UPLC-LTQ-Orbitrap-MS). Methods Warters ACQUITY UPLC HSS T3 (2.1 mm ×100 mm, 1.8 μm) was used as chromatographic column with mobile phase: 0.1% formic acid water (A)-0.1% formic acid acetonitrile (B) with gradient elution, and flow rate was 0.3 ml/min. Electrospray ion source (ESI) and an electrostatic field orbital ion trap mass analyzer were adopted, which was used to collect mass spectrometry fragment information with positive and negative ion modes, by comparing with the relative retention time of the reference substance. In addition, the fragment information of the mass spectrum was used to identify the compounds. The accurate identification of the chemical components in Shenqi Dihuang decoction was confirmed with literature. Results The study found that UPLC-LTQ-Orbitrap-MS technology could be used to identify 62 chemical components, including 13 aromatic acids, 9 flavonoids, 8 saponins, and 5 aromatic amines, 3 keto acids, 2 phenols, 1 aromatic quinone and other ingredients in Shenqi Dihuang decoction. Conclusion The identification analysis method in this study was efficient and accurate, which could be applied to the identification and analysis of chemical components in Shenqi Dihuang decoction and provided the important experimental data for the research on the material basis and mechanism.

Objective:To explore the feasibility of using multivariate statistical analysis to evaluate the quality of Astmgali radix from different habitats. Methods:The contents of Astragaloside saponinⅠ, Astragalus saponin Ⅱ, Astragaloside Ⅲ and Astragaloside A were determined by UPLC-ELSD. The components of astragalus saponins from different habitats were analyzed by TOPSIS and cluster thermogram.Results:TOPSIS analysis showed that the comprehensive quality of Astmgali radix samples from Shanxi, Gansu and Inner Mongolia was related(were 0.297 3, 0.346 0, 0.322 5), and the whole quality of S13 and S14 from Shanxi and N5 from Inner Mongolia were higher than others. Cluster thermogram showed that Astmgali radix was grouped into three groups according to region, and the quality difference of Astmgali radix was mainly reflected on Astragaloside saponinⅠand Astragalus saponin Ⅱ. Conclusions:The theory of multivariate statistical analysis is perfect, objective and reliable. It can be used as a reference for comprehensive quality evaluation of Traditional Chinese Medicine (TCM), selection of excellent germplasm resources and traceability of origin of TCM.

Grape (Vitis vinifera L.) in production is frequently exposed to inadequate light, which significantly affects its agronomic traits via inhibiting their physiological, metabolic and developmental processes. To explore the mechanism how the grape plants respond to the weak light stress, we used 'Yinhong' grape and examined their physiology-biochemistry characteristics and transcriptional profile under different levels of weak light stress. The results showed that grape seedlings upon low intensity shading treatments were not significantly affected. As the shading stress intensity was strengthened, the epidermis cells, palisade tissue, and spongy tissue in the leaves were thinner, the intercellular space between the palisade tissue and spongy tissue was larger compared with that of the control, and the activities of superoxide dismutase, catalase and peroxidase were decreased gradually. Additionally, the soluble protein content increased and the free proline content decreased gradually. Compared with the control, significant changes in plant photosynthetic characteristics and physiology-biochemistry characteristics were observed under high intensity of shading (80%). RNA-seq data showed that the differentially expressed genes between CK and T2, CK and T4, T2 and T4 were 13 913, 13 293 and 14 943, respectively. Most of the enrichment pathways were closely related with the plant's response to stress. Several signaling pathways in response to stress-resistance, e.g. JA/MYC2 pathway and MAPK signal pathway, were activated under weak light stress. The expression level of a variety of genes related to antioxidation (such as polyphenol oxidase and thioredoxin), photosynthesis (such as phytochrome) was altered under weak light stress, indicating that 'Yinhong' grape may activate the antioxidation related pathways to cope with reactive oxygen species (ROS). In addition, it may activate the expression of photosynthetic pigment and light reaction structural protein to maintain the photosynthesis activity. This research may help better understand the relevant physiological response mechanism and facilitate cultivation of grape seedlings under weak light.
Vitis/metabolism* ; Gene Expression Regulation, Plant ; Photosynthesis/genetics* ; Plant Leaves ; Light ; Seedlings/metabolism*

Objective To explore the predictive value of the expression of CD44v6 and EGFR on the efficacy of neoadjuvant chemotherapy (NACT) in stageⅡ-Ⅲ cervical cancer. Methods A total of 53 patients with stageⅡ-Ⅲ cervical cancer diagnosed by pathology were selected. All patients received two cycles of paclitaxel+platinum NACT. The pathological tissue samples of cervical tumors before NACT treatment were collected. The expression of CD44v6 and EGFR were detected by the immunohistochemical SP method, and we analyzed their predictive value of NACT in stageⅡ-Ⅲ cervical cancer. Results Among the 53 patients, 38 were in the NACT effective group (CR+PR), and 15 were in the NACT ineffective group (SD+PD). The expression of CD44v6 in the ineffective group was significantly higher than that in the effective group (P < 0.05). The expression of CD44v6 was significantly different in patients with CR, PR, and SD (P < 0.05). The AUC of CD44v6 to NACT effect on stage Ⅱ-Ⅲ cervical cancer was 0.74 (P < 0.05). The patients in the high expression group of CD44v6 had worse efficacy in NACT than those in the low expression group of CD44v6 (P < 0.05). Pearson test showed that CD44v6 and EGFR expression were correlated (R=0.34, P < 0.05). Conclusion High expression of CD44v6 may reduce the efficacy of NACT in stageⅡ-Ⅲ cervical cancer, suggesting that the expression of CD44v6 has a certain predictive value and clinical significance in the efficacy of paclitaxel+platinum NACT on cervical cancer. Moreover, CD44v6 is positively correlated with EGFR expression.

Objective:To analyze the clinical characteristics and causes of death of 80 dead cases with confirmed coronavirus disease 2019 (COVID-19).Methods:The clinical data of 80 dead patients with COVID-19 who were admitted to Renmin Hospital of Wuhan University from January 11 to February 11, 2020 were retrospectively analyzed.The laboratory examination indexes (including white blood cells, lymphocytes, procalcitonin (PCT), lactic acid, D-dimmer, fibrinogen degradation products, N-terminal pro-brain natriuretic peptide (N-proBNP), ultra sensitive-troponin I, lactate dehydrogenase (LDH) and CD4 + T lymphocyte) of the patients at the time of admission were compared with the indexes at the last time before death. Statistical analysis was conducted by using paired t test or Wilcoxon′s signed rank test. Results:The median age was 72 years old of the 80 patients, and 78.75%(63/80) of them were older than 60 years. Thirty-six cases (45.00%) were severe and 44(55.00%) were critical at admission. Fifty-eight cases (72.50%) had underlying diseases. The common underlying diseases were hypertension, diabetes mellitus, coronary atherosclerotic heart disease, and chronic obstructive pulmonary disease. Comparing the patients′ first laboratory tests at admission with those before death, white blood cells increased (8.01(4.86, 12.29)×10 9/L vs 12.55(8.25, 17.66)×10 9/L), lymphocytes decreased (0.70(0.46, 0.88)×10 9/L vs 0.54(0.39, 0.75)×10 9/L), PCT increased (0.20(0.11, 0.74) μg/L vs 1.00(0.20, 1.99) μg/L), lactic acid increased (2.10(1.40, 3.10) mmol/L vs 3.10(2.60, 4.10) mmol/L), D-dimmer increased (4.33(0.97, 18.98) mg/L vs 15.29(5.17, 53.44) mg/L), fibrinogen degradation products increased (15.90(3.58, 76.60) mg/L vs 63.14(21.23, 110.67) mg/L), N-proBNP increased (1 078.00(347.35, 2 996.50) ng/L vs 3 439.50(1 576.00, 9 281.50) ng/L), ultra-sensitive troponin I increased (0.08(0.03, 0.17) μg/L vs 0.33(0.14, 2.47) μg/L), LDH increased (397.00(327.00, 523.50) U/L vs 624.00(481.00, 854.00) U/L) and CD4 + T lymphocyte decreased (137.00(104.00, 168.00)/μL vs 97.00(67.00, 128.00)/μL). The differences between the two groups were all statistically significant ( W=238.00, 1 053.50, 150.00, 152.00, 192.00, 190.00, 108.00, 57.00, 53.00 and 40.00, respectively, all P<0.05). All patients received antiviral and respiratory-support therapy and the main cause of death was respiratory failure caused by intractable hypoxemia and multiple organ failure. Among them, seven cases died in one day hospitalization, and 66 cases died in seven days hospitalization. Conclusions:Elderly patients with a variety of chronic underlying diseases have poor prognosis. It′s essential to pay more attention and deal with the above clinical characteristics at an early stage to improve the outcome of the COVID-19 patients.

Objective To collect the serum samples of patients with nonalcoholic fatty liver disease (NAFLD), and to investigate the changes in serum metabolic biomarkers before and after lifestyle intervention. Methods A total of 23 patients who were diagnosed with NAFLD in Department of Gastroenterology and Inpatient Department, Putuo District Central Hospital of Shanghai, from January 2019 to January 2020 were enrolled, and all patients received the intervention with aerobic exercise and equicaloric low-carbohydrate high-protein diet. A total of 13 healthy volunteers who underwent physical examination in Physical Examination Center were enrolled as control group. For the patients with NAFLD, basic information was collected before and after intervention, blood samples were collected twice to measure liver function, blood glucose, and blood lipids, and part of serum was used for serum metabolomics analysis. The serum samples were analyzed by ultra-performance liquid chromatography/tandem high-resolution mass spectrometry. The data collected were processed in Compound Discover, and then principal component analysis (PCA) and orthogonal partial least squares discriminant analysis were used to establish the profile of differentially expressed blood metabolites between patients and healthy people and perform the enrichment analysis of differentially expressed metabolic pathways. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon non-parametric test was used for comparison of non-normally distributed continuous data between two groups. Results After lifestyle intervention, the patients had significant reductions in body mass index ( P < 0.01), body weight ( P < 0.01), and serum biochemical parameters alkaline phosphatase, albumin, gamma-glutamyl transpeptidase, and alanine aminotransferase (all P < 0.05), as well as a significant reduction in total protein ( P < 0.01), while there were no significant improvements in cholinesterase, aspartate aminotransferase, and glucose. As for the four items for blood lipids, there was a significant reduction in triglyceride ( P < 0.01), while there were no significant improvements in high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and total cholesterol. The metabolomics analysis showed that 33 serum metabolites changed significantly after lifestyle intervention. In addition, PCA results showed that after intervention, the level of metabolites in patients tended to be normal. The signaling pathway analysis showed that exercise and diet mainly affected the pathways of bile acid, unsaturated fatty acid synthesis, and phenylalanine metabolism. Conclusion Lifestyle intervention can achieve varying degrees of reduction in the body weight of patients with NAFLD, improve serum biochemical parameters, and regulate the abnormal metabolic pathway in patients with NAFLD, which has important clinical value and significance for guiding clinicians to formulate reasonable diet and exercise strategies for patients with NAFLD and prevent the progression of NAFLD.