Bioinformatics analysis and key gene verification of sepsis myocardial macrophage microarray data based on GEO database / 中华心血管病杂志

Objective: Bioinformatics analysis was used to screen differentially expressed genes (DEGs) in macrophages of sepsis myocardial injury and to verify key genes. Methods: Experiment 1 (gene chip and bioinformatics analysis): The gene chip data GSE104342 of cardiac macrophages in septic mice was downloaded from Gene Expression Omnibus database. DEGs were obtained by R language analysis. DAVID online database was used to obtain gene ontology and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of DEGs. STRING online database was used for protein-protein interaction network analysis of DEGs, and then key genes were screened by using Cytoscape software and molecular complex detection (MCODE) plug-ins. Experiment 2 (sepsis model construction and related protein verification): Ten male C57BL/6 mice, aged 8-14 weeks. Five mice were randomly selected as control group, and 5 mice were selected as the sepsis group by building a mice sepsis model in vivo. Echocardiography was used to detect the cardiac function. Hematoxylin-eosin staining was used to assess the cardiac morphology. TUNEL staining was used to evaluate cardiomyocyte apoptosis. Immunofluorescence staining was used to detect the expression of differentiation antigen cluster 206 (CD206),inducible nitric oxide synthases (iNOS),F4/80,suppressor of cytokine signaling 3 (Socs3) ,interleukin 1 receptor antagonist (Il1rn) and chemokine C-C motif ligand 7 (Ccl7) protein. RAW264.7 macrophages were cultured in vitro and divided into 2 groups: LPS groupstimulated by lipopolysaccharide (LPS, 1 mg/L) and blank control group treated with equal-volume phosphate buffer solution. Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the expression of Socs3, Il1rn and Ccl7 in vitro. Results: Experiment 1: 24 647 genes were screened in GSE104342 dataset and 177 genes (0.72%) were differential expression, including 120 up-regulated genes and 57 down-regulated genes. Gene ontology enrichment analysis showed that DEGs were mainly involved in inflammatory response, immune response, apoptosis regulation and antigen processing and presentation. KEGG signaling pathway analysis showed that DEGs in cardiac macrophages of septic mice were mainly enriched in cytokine-cytokine receptor interaction, tumor necrosis factor signaling pathway, NOD like receptor signaling pathway. Three hub genes were obtained by STRING and Cytoscape analysis, including Socs3, Il1rn and Ccl7. Experiment 2: In vivo, it was found that compared with the control group, the cardiac function of the sepsis mice decreased significantly, the myocardial cells were significantly edema, inflammatory cell infiltration, myocardial fiber rupture, some myocardial nuclei dissolved and disappeared, and the cardiomyocyte apoptosis increased, suggesting that the sepsis myocardial injury model of mice was successfully constructed. Compared with the control group, the expression of CD206 in the myocardium of septic mice was down-regulated, the expression of iNOS, F4/80, Socs3, Il1rn and Ccl7 were up-regulated. In addition, there was co-localization between Socs3, Il1rn, Ccl7 and F4/80 protein. Compared with the blank control group, the expression of Socs3, Il1rn and Ccl7 significantly upregulated after LPS intervention in vitro by RT-PCR. Conclusions: The selected key genes Socs3, Il1rn and Ccl7 were up-regulated in myocardial macrophages of septic mice. Socs3, Il1rn and Ccl7 are expected to become new targets for the diagnosis and treatment of sepsis cardiac injury.

Independent and joint association of physical activity and sedentary behavior on all-cause mortality / 中华医学杂志(英文版)

BACKGROUNDS@#Physical activity (PA) and sedentary behavior (SB) have been associated with mortality, while the joint association with mortality is rarely reported among Chinese population. We aimed to examine the independent and joint association of PA and SB with all-cause mortality in southern China.@*METHODS@#A cohort of 12,608 China Hypertension Survey participants aged ≥35 years were enrolled in 2013 to 2014, with a follow-up period of 5.4 years. Baseline self-reported PA and SB were collected via the questionnaire. Kaplan-Meier curves (log-rank test) and Cox proportional hazards regression were performed to evaluate the associations of PA and SB on all-cause mortality.@*RESULTS@#A total of 11,744 eligible participants were included in the analysis. Over an average of 5.4 years of follow-up, 796 deaths occurred. The risk of all-cause mortality was lower among participants with high PA than those with low to moderate level (5.2% vs. 8.9%; hazards ratio [HR]: 0.75, 95% confidence interval [CI]: 0.61-0.87). Participants with SB ≥ 6 h had a higher risk of all-cause mortality than those with SB <6 h (7.8% vs. 6.0%; HR: 1.37, 95% CI: 1.17-1.61). Participants with prolonged SB (≥6 h) and inadequate PA (low to moderate) had a higher risk of all-cause mortality compared to those with SB < 6 h and high PA (11.2% vs. 4.9%; HR: 1.67, 95% CI: 1.35-2.06). Even in the participants with high PA, prolonged SB (≥6 h) was still associated with the higher risk of all-cause mortality compared with SB < 6 h (7.0% vs. 4.9%; HR: 1.33, 95% CI: 1.12-1.56).@*CONCLUSIONS@#Among Chinese population, PA and SB have a joint association with the risk of all-cause mortality. Participants with inadequate PA and prolonged SB had the highest risk of all-cause mortality compared with others.

Quantitative detection of the KRAS gene mutation in circulating tumor DNA for patients with colorectal cancer / 基础医学与临床

Objective To establish a liquid biopsy technique of KRAS gene G12D mutation and to assess its diagnostic value. Methods KRAS G12D mutation was analyzed by ddPCR in plasma DNA from 52 colorectal cancer patients and compared that of to 80 healthy subjects. KRAS gene sequencing in cancerous tissue of colorectal cancer patient being set as a golden standard, we evaluated the accuracy of ddPCR and analyzed the correlation between G12D mutation rate, plasma concentration;and their clinical manifestations in CRC. Results ddPCR indicated that KRAS G12D mutation rate and concentration(26.92％, 81.5 copies/mL) in the plasma samples of colorectal cancer patients were significantly higher than that of healthy subjects (8.75％, 16 copies/mL). Colorectal cancer patients with highly differentiated adenocarcinoma showed a significantly higher number of mutant copies than medium and low differentiated adenocarcinoma(P＜0.05);M2 patients had a significantly higher number of mutant copies than N1 and NO patients (P＜0.05);The concordance rate of KRAS gene mutation between cancerous tissue and plasma ctDNA was 87.50％ in CRC.Conclusions ddPCR is a fast, noninvasive and accurate method for plasma testing of ctDNA, and the test results could be used to monitor the course of the disease and as clinical guidelines.

Age-specific detection rates of colorectal neoplasms by colonoscopic screening in high-incidence rural area / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare the age-specific detection rates of colorectal neoplasms by colonoscopic screening among high-risk population in rural area and to provide evidence for determining the initial age in the colorectal cancer screening.</p><p><b>METHODS</b>The age-specific detection rates of colorectal cancers and adenomas, and the proportion of detected cases in each age group after screening by the optimized sequential colorectal cancer screening program in Jiashan County, Zhejiang Province, were analyzed.</p><p><b>RESULTS</b>Mass screening with the optimized sequential colorectal cancer screening program in Jiashan was conducted and 8867 colonoscopic examinations were performed. A total of 1811 individuals with at least one colorectal neoplastic lesion were found. Among them, there were 92 (1.04%) colorectal cancer patients, 1164 (13.13%) patients with at least one adenoma and 377 (4.25%) patients with at least one advanced adenoma. The detection rate of colorectal neoplastic lesions for the age group 40 - 44 was considerably lower than that in the age group 45 - 49. Hypothetically, to increase the initial age to 45-years for the optimized sequential colorectal cancer screening program would yield a 7.84% increase in the detection rate of total colorectal neoplastic lesions, 13.46% increase in the detection rate of colorectal cancer, 8.76% increase in the detection rate of adenoma, 12.24% increase in the detection rate of advanced adenoma and 19.64% lower in the cost of initial screening, 13.30% lower in the cost of colonoscopic screening.</p><p><b>CONCLUSIONS</b>Among the high-risk population aged 40 to 74, the detection rates of colorectal cancer and adenoma are increasing with the increase of age, the detection rates of colorectal cancer and adenoma as well as proportion of detected cases in the 40 - 44 age group are considerably lower. To increase the initial age to 45 for colorectal cancer screening in rural area of China is reasonable.</p>

Association of HLA-G 14bp gene polymorphisms and plasma sHLA-G level with susceptibility to EV71 infection in children / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Explore the relationship between the HLA-G 14bp insertion/deletion polymorphism and the infection of Enterovirus 71 (EV71) for children.</p><p><b>METHODS</b>We genotyped HLA-G 14bp insertion/deletion polymorphism of 125 severe HFMD children infected with EV71 and 133 normal controls by PCR-PAGE;detected the plasma sHLA-G level of 66 heavy type and 15 critical type and 133 normal controls by ELISA.</p><p><b>RESULTS</b>Frequencies of the genotype 14 bp - / - ,14 bp + / - and 14 bp + / + were 49.6% , 42.4% and 8.0% for the severe HFMD children infected with EV71, and 34.6%, 48.9% and 16.5% for the normal controls, respectively. A significant difference was observed for the frequencies of the HLA-G 14bp genotype between the two groups(chi2 = 7.850, P = 0.020). And for the allele frequencies. The plasma sHLA-G levels in heavy type were dramatically higher than that in normal controls (Z = -9.692, P = 0.000). The plasma sHLA-G levels in children with critical HFMD were dramatically higher than that with heavy type (Z = -2.420, P = 0.016).</p><p><b>CONCLUSION</b>There was a relationship between the HLA-G 14 bp insertion/deletion polymorphism and the susceptibility to the severe HFMD children infected with EV71 and the plasma sHLA-G might be considered as a index for auxiliary diagnosis the severe HFMD infected with EV71.</p>

Optimization of sequential screening scheme in prevention of colorectal neoplasm / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To improve early diagnosis rate and reduce the incidence rate of colorectal cancer, through the application of optimized sequential screening scheme for colorectal neoplasm in general population.</p><p><b>METHODS</b>Quantitative risk assessment by questionnaires survey and fecal occult blood test (FOBT) were used to proceed preliminary screening among people aged 40 to 74. Electronic colonoscopy was applied to examine the whole colon and rectum among identified high-risk subjects. The detected cases received treatment for colorectal cancer, adenomatous polyps or non-adenomatous polyps. The early diagnosis rate and incidence rate of colorectal cancer were evaluated and compared with those before screening.</p><p><b>RESULT</b>With application of optimized sequential screening schemes, various types of colorectal lesions were detected in 1 117 subjects, including 69 cases of colorectal cancer, 701 cases of colorectal adenoma and 211 cases of advanced adenoma. The early diagnosis rate of colorectal cancer was increased by 58.19%, and its incidence rate also decreased significantly.</p><p><b>CONCLUSION</b>The optimized sequential screening scheme is simple, economical, efficient in colorectal cancer screening of general population.</p>

Determination of two saponins in Tupistra chinensis rhizomes by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develog an HPLC method determination of two new saponins in the dried rhizomes of Tupistra chinensis.</p><p><b>METHOD</b>The analysis was performed on YMC-Pack ODS-AQ (4.6 mm x 250 mm, 5 microm) eluted with of acetonitrile and waterin gradient mode. The conentration of acetonitrile in the mobile phase changes from 10% to 100% within 85 minutes. The detection wavelength was set at 203 nm. The flow rate was 1.00 mL x min(-1) and column temperature was set at 30 degrees C.</p><p><b>RESULT</b>The linear relationships of two authentic saponins were determined within the range from 7.55 microg to 60.40 microg (r=0.9996) and 8.00 microg to 48.00 microg (r=0.9996), respectively. Using the method above, the contents of two saponins were determined as 0.261% and 0.242%, with the recoveries as 98.6% and 98.3% with RSD 1.7% and 1.7%, respectively.</p><p><b>CONCLUSION</b>A convenient and reliable method was developed to determine the content of two saponins in the dried rhizomes of Tupistra chinensis.</p>

Effects of sodium hyaluronate on growth and adhesion of two human colorectal cancer cell lines in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of sodium hyaluronate on the growth and adhesion of colorectal cancer cells.</p><p><b>METHODS</b>Human colorectal cancer cell lines SW620 and Colo205 were treated with sodium hyaluronate (25 -2,500 microg/ml), and cancer cell proliferation was measured by MTT assay in vitro. Flow-cytometric analysis was applied to detect expression of CD44 on SW620 and Colo205 cells.</p><p><b>RESULT</b>In vitro sodium hyaluronate enhanced proliferation of Colo205 cells, but it had no appreciable effect on SW620 growth under the same doses, Meantime, CD44 expression on cancer cells decreased compared with controls.</p><p><b>CONCLUSION</b>In vitro sodium hyaluronate has different effects on growth of different colorectal cancer cell lines, but can inhibit CD44 expression of colorectal cancer cells and influence their ability of adhesion.</p>