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1.
Article in Chinese | WPRIM | ID: wpr-935250

ABSTRACT

The aim of this study was to construct a simple, rapid and ultra-sensitive optical biosensing technique based on rolling circle amplification (RCA), and to apply it to multiple detection of drug-resistant genes of mycobacterium tuberculosis. The common mutation sites of isoniazid, rifampicin and streptomycin resistance genes are katG315 (AGC➝ACC), rpoB531 (CAC➝TAC) and rpsL43 (AAG➝AGG). For these three gene sites, from February 2020 to May 2021, in the Department of Laboratory Medicine of the First Affiliated Hospital of Army Military Medical University, the padlock probe (PLP), primers and capture probes were designed. And a solid-phase RCA constant temperature amplification reaction system based on magnetic beads was constructed and the experimental parameters were optimized. The RCA products were accurately captured by the multicolor fluorescent probes (Cy3/Cy5/ROX), and the single-tube multiple detection of three mutation genes was realized. The sensitivity, specificity and linear range of this method were further verified. The results showed that the response range of katG315 in the same reaction system ranged from 1.0 pmol/L to 0.1 nmol/L. The response range of rpoB531 and rpsL43 ranged from 1.0 pmol/L to 50.0 pmol/L and 1.0 pmol/L to 20.0 pmol/L, and the method had good specificity and sensitivity, and could accurately identify single base mutations in mixed targets, with the minimum detection limit as low as 1.0 pmol/L. The recoveries of simulated serum samples were 95.0%-105.2%. In conclusion, the constant temperature amplification multiple detection method constructed in this study can quickly realize the single-tube multiple detection of three drug resistance mutation sites. This technology is low-cost, simple and rapid, and does not rely on large equipment, providing a new analysis method for pathogen drug resistance gene detection.


Subject(s)
Drug Resistance , Fluorescent Dyes , Humans , Mycobacterium tuberculosis/genetics , Nucleic Acid Amplification Techniques
2.
Article in Chinese | WPRIM | ID: wpr-942365

ABSTRACT

Objective To evaluate the molluscicidal effect of 50% wettable powder of niclosamide ethanolamine salt (WPNES) against Oncomelania hupensis on the soil surface and inside the soil layer by immersion method in winter. Methods O. hupensis snails were placed on the soil surface and 2, 5 cm and 10 cm under the soil layer outdoors in winter, and then immersed in 50% WPNES at concentrations of 1 mg/L and 2 mg/L for 1, 3 d and 7 d, while dechlorinated water served as controls. Snail mortality was observed following immersion with 50% WPNES on the soil surface and inside the soil layer. Results Following immersion with 50% WPNES at concentrations of 2 mg/L and 1 mg/L outdoors in winter, the 3-day corrected snail mortality rates were 98.0% and 76.0% on the soil surface, and the 7-day corrected snail mortality rate was both 100.0%. Following immersion with 50% WPNES at concentrations of 2 mg/L and 1 mg/L outdoors in winter, the 7-day corrected snail mortality rates were 95.5% and 85.6% 2 cm below the soil layer, 66.0% and 6.4% 5 cm below the soil layer. However, the 7-day snail mortality rate swere comparable between the 50% WPNES treatment group (at 2 mg/L and 1 mg/L) and controls 10 cm below the soil layer (both P > 0.05). Conclusion Immersion of 50% WPNES at a concentration of 2 mg/L for 7 days presents a high molluscicidal efficacy against O. hupensis on the soil surface and 5 cm within the soil layers in winter.

3.
Article in English | WPRIM | ID: wpr-880686

ABSTRACT

OBJECTIVES@#To determine the association between glycosylated hemoglobin (HbA1c) and ambulatory blood pressure or heart rate in hypertensive patients.@*METHODS@#A total of 585 patients, who performed ambulatory blood pressure monitoring (ABPM) from September 2018 to April 2019 in Xiangya Hospital, Central South University, were enrolled and assigned into 2 groups (470 in a hypertensive group and 115 in a normal group). HbA1c levels were compared. According to the HbA1c level, the hypertensive group was divided into 2 subgroups: A high HbA1c group (HbA1c≥6.1%, @*RESULTS@#The hypertensive group had higher HbA1c level than the normal group [(6.1±1.3)% vs (5.1±1.7)%, @*CONCLUSIONS@#In hypertensive patients, HbA1c is positively correlated with ambulate blood pressure, blood pressure load, and heart rate, and it has no correlation with blood pressure variability, heart rate variability, or morning blood pressure.


Subject(s)
Blood Pressure , Blood Pressure Monitoring, Ambulatory , Glycated Hemoglobin A/analysis , Heart Rate , Humans , Hypertension
4.
Acta Pharmaceutica Sinica ; (12): 574-581, 2019.
Article in Chinese | WPRIM | ID: wpr-780135

ABSTRACT

Tropinone reductase I (TRI) is a key branch point enzyme in the midstream of tropane alkaloids (TAs) biosynthesis pathway and represents an important target for TAs metabolic engineering, which can lead to metabolic flux of substrate tropinone to TAs. A novel TRI gene was isolated from Datura arborea, a woody resource plant, and designated as DaTRI2 (GenBank accession number is MH705164). The full-length cDNA of DaTRI2 with 1 135 bp exhibits a high sequence homology (96.8%) with DaTRI, and is predicted to encode a protein of 347 amino acids. Deduced DaTRI2 protein contain a conserved TGXXXGXG motif involved in NADPH binding, the catalytic N-S-Y-K tetrad motif and eleven amino acid residues important for binding to its substrate tropinone. The phylogenetic analysis revealed that DaTRI2 and other TRIs from Solanaceous plants belong to the same cluster and DaTRI2 exhibited closest phylogenetic proximity to TRIs from Datura. DaTRI2 was expressed in E. coli and the purified recombinant protein can catalyze both tropinone reduction and tropine oxidation with an optimum pH value of 8.0 and 9.6, respectively. When tropinone was used as the substrate, the Km and Vmax values of DaTRI2 at pH 6.4 were 210.05 μmol·L-1 and 69.6 nkat·mg-1 protein respectively, while the Km and Vmax values for tropine as the substrate were 188.03 μmol·L-1 and 114 nkat·mg-1 protein respectively, at pH 9.6. DaTRI2 transcript was most abundant in the young leaf, followed by the root. Cloning of DaTRI2 gene and biochemical analysis of recombinant DaTRI2 facilitate further research on the molecular mechanism on TAs biosynthesis in woody plants and provide a more potent candidate for TAs metabolic engineering.

5.
Article in Chinese | WPRIM | ID: wpr-777916

ABSTRACT

Objective To understand the current epidemic and immunity status of hepatitis B virus in Ma’anshan City, and to compare the prevention and control effect after the adjustment of hepatitis B vaccine immunization strategy. Methods Multi-stage random sampling method was used to select 10 investigation points in the whole city, a random sample of 3 460 people under 60 years old was included according to urban and rural stratification. questionnaires and blood were collected from the subjects, and domestic enzyme linked immunosorbent assay (ELISA) was used for hepatitis B immunoglobulins detection. Results The total positive rate of hepatitis B virus surface antigen (HBsAg), hepatitis B virus surface antibody (HBsAb) and Hepatitis B virus (HBV) infection was 3.32%, 51.21% and 29.16% respectively. The positive rate of HBsAb in urban area was higher than that in rural area ( 2=28.204, P<0.001). The positive rate of HBsAb was significantly different between the medical and nursing staff and other occupational groups ( 2=22.772, P<0.001). The difference of HBsAb positive rate before and after HepB vaccine content adjustment was statistically significant ( 2=90.331, P<0.001). The rate of HepB decreased with age ( 2trend=1 984.342, P<0.001). Conclusions Since HepB was incorporated into the immunization program, hepatitis B prevention and control in school-age children has achieved remarkable results. More attention should be paid on the low positive rate of HBsAb in students and the low immunization rate of HepB in adults.

6.
Article in Chinese | WPRIM | ID: wpr-665941

ABSTRACT

Objective To observe the effects of constant light exposure on the obesity in high fat diet rats. Methods Thirty-two male SD rats were randomly divided into four groups:rats on a normal chow exposed to standard light-dark cycle ( group A) , rats on a normal chow exposed to constant light ( group B) , rats on a high fat diet exposed to standard light-dark cycle ( group C) , and rats on a high fat diet exposed to constant light ( group D) . Body weights and food intakes were recorded weekly throughout the 12-week study. Body weight, fat mass, visceral adipose tissue weight, intraperitoneal glucose tolerance test ( IPGTT) results, insulin resistance parameters, serum lipids and levels of interleukin 6 (IL-6), tumor necrosis factor-α(TNF-α) were compared among groups. Epididymal adipose tissues mRNA expression of circadian clock genes, i. e. clock, bmal1, rorα, rev-erbα, cry1, per1, and per2 were analyzed by realtime PCR. Results From the 9th week, body weights of rats in group D were significantly higher than those in group C (all P<0. 05). At the 12th week, area under curve of IPGTT (AUC-IPGTT) in groups B, C, and D were significantly higher than that in group A. AUC-IPGTT in group D was significantly higher than that in group C (all P<0.05). Compared with group C,asignificant increase in fat mass,visceral adipose tissue weight,homeostasis model assessment for insulin resistance, serum cholesterol, TNF-α levels were observed in group D ( all P<0. 05). And a significant decrease in quantitative insulin sensitivity check index ( QUICKI) and high density lipoprotein-cholesterol were observed in group D in comparison with group C (both P<0. 05). Circadian clock genes (clock, rorα, rev-erbα, cry1, per1) mRNA expressions in group B and D were significantly different from those in group A (all P<0. 05) . Expression of cry1 in group D was significantly higher than that in group C. In group C, rev-erbαmRNA expression was significantly down-regulated in comparison with group A (P<0. 05). Conclusion Constant light exposure exaggerates obesity, glycolipid metabolism abnormality, inflammation, and insulin resistance in high fat diet rats.

7.
Acta Pharmaceutica Sinica ; (12): 1913-2016.
Article in Chinese | WPRIM | ID: wpr-779351

ABSTRACT

Atropa belladonna L. is the commercial plant material for production of tropane alkaloids, including hyoscyamine and scopolamine. The wild-type Atropa belladonna is characterized by the hyoscyaminerich chemotype, in which the hyoscyamine content is much higher than the scopolamine content. It is the common goal for the pharmaceutical industry to increase the content of scopolamine in A. belladonna. Based on the T0 progeny of transgenic A. belladonna with NtPMT and HnH6H overexpression, T1 progeny of transgenic A. belladonna were obtained through self-pollination and used in a field trial. The 461 bp fragment of NtPMT and the 1 077 bp HnH6H were simultaneously expressed from T1 progeny of transgenic A. belladonna, but were not obtained from the wild-type A. belladonna. At the transcription level, the expression of NtPMT and HnH6H were detected in T1 progeny of transgenic A. belladonna, but were not detected in the wild-type plants. Further, the alkaloids were analyzed by HPLC. In the stems and leaves of T1 progeny of transgenic A. belladonna, hyoscyamine was not detected and scopolamine was detected at very high levels; in the stems and leaves of wild-type A. belladonna, hyoscyamine was detected at much higher levels. In the leaves of T1 progeny of transgenic A. belladonna, the content of scopolamine was 15-36 folds higher than that of wildtype leaves; in the stems of T1 progeny of transgenic A. belladonna, the scopolamine content was 37-108 folds higher than that of wild-type stems. In conclusion, overexpression of NtPMT and HnH6H greatly enhanced conversion of hyoscyamine into high-value scopolamine and improved the commercial value of A. belladonna.

8.
Article in Chinese | WPRIM | ID: wpr-814997

ABSTRACT

OBJECTIVE@#To explore the effect of ROCK inhibitor Y-27632 on the matrix metalloproteinase 2 and 9 (MMP2 and MMP9) gene expression and activity in tumor necrosis factor α (TNF-α)-treated human umbilical vein endothelial cell (HUVEC).
@*METHODS@#HHUVEC was divided into 3 groups, a control group, a TNF-α group, and a TNF-α plus Y-27632 group. The expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), MMP2 and MMP9 were examined by real-time PCR. The MMP2/9 activity was measured by gelatin zymography.
@*RESULTS@#Compared to the control group, the mRNA expressions of ICAM-1, VCAM-1, MMP2 and MMP9 were increased TNF-α-treated cells, which were suppressed by ROCK inhibitor (P<0.01). The MMP2/9 activity was elevated in TNF-α-treated cells, which was reversed by ROCK inhibitor (P<0.05).
@*CONCLUSION@#ROCK inhibitor can suppress TNF-α-induced inflammation in endothelial cells through down-regulation of MMP2/9.


Subject(s)
Amides , Cells, Cultured , Down-Regulation , Endothelial Cells , Humans , Intercellular Adhesion Molecule-1 , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Pyridines , Signal Transduction , Tumor Necrosis Factor-alpha , Umbilical Veins , Vascular Cell Adhesion Molecule-1 , rho-Associated Kinases
9.
Acta Pharmaceutica Sinica ; (12): 1346-55, 2015.
Article in Chinese | WPRIM | ID: wpr-505057

ABSTRACT

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

10.
Acta Pharmaceutica Sinica ; (12): 1346-1355, 2015.
Article in Chinese | WPRIM | ID: wpr-320078

ABSTRACT

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.


Subject(s)
Cloning, Molecular , DNA, Complementary , Datura , Genetics , Escherichia coli , Hyoscyamine , Chemistry , Mixed Function Oxygenases , Genetics , Plant Leaves , Plant Roots , Recombinant Proteins , Genetics , Scopolamine , Chemistry
11.
Article in Chinese | WPRIM | ID: wpr-815505

ABSTRACT

OBJECTIVE@#To examine the serum levels of S100 calcium-binding protein A8/A9 complex (S100A8/ A9) in patients with acute coronary syndrome (ACS) and to explore the relation between the serum levels of S100A8/A9 and the degree of coronary lesion.@*METHODS@#A total of 126 patients with coronary heart disease were enrolled from Xiangya Hospital of Central South University between September 2010 and January 2011, which included 51 patients with unstable angina pectoris (UAP group, n=51), 50 patients with acute myocardial infarction (AMI group, n=50), and 25 patients with stable angina pectoris (SAP group, n=25). Twenty-five healthy volunteers were served as a normal control group (NC group, n=25). According to the coronary artery lesion area, ACS patients were also divided into a single-branch group, a double-branch group and a triple-branch group. Serum levels of S100A8/A9 were measured by enzyme-linked immunosorbent assay on the day when the patients admitted to the hospital and on the day after one-week treatment (UAP group + AMI group). The serum levels were compared among the various branch groups. The short-term prognosis in patients with ACS was investigated by phone follow-up after 3 months.@*RESULTS@#1) The S100A8/A9 level in the SAP group was significantly higher than that in the normal control group (P0.05); 2) After one-week standard treatment, the serum levels of S100A8/A9 in patients with ACS were significantly reduced compared with that at the admission (P 0.05).@*CONCLUSION@#The serum level of S100A8/A9 is significantly elevated in patients with ACS, which might be positively correlated with the number of the coronary lesion branches.


Subject(s)
Acute Coronary Syndrome , Blood , Pathology , Angina Pectoris , Angina, Unstable , Coronary Artery Disease , Enzyme-Linked Immunosorbent Assay , Humans , Leukocyte L1 Antigen Complex , Blood , Prognosis
12.
Article in Chinese | WPRIM | ID: wpr-319655

ABSTRACT

Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.


Subject(s)
Alkaloids , Genetics , Metabolism , Atropa belladonna , Genetics , Metabolism , Gene Expression Regulation, Plant , Genetics , Hyoscyamine , Genetics , Metabolism , Plants, Medicinal , Genetics , Metabolism , Scopolamine , Metabolism , Tropanes , Metabolism
13.
Article in Chinese | WPRIM | ID: wpr-318624

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the biological activity of polysaccharide of Cipangopaludina chinensis (PCC) against HBV in vitro.</p><p><b>METHOD</b>HepG2 2. 2. 15 cells were taken as the in vitro experimental model. The cell toxicity was observed by MTT. PCC of different safe concentrations and positive control medicine 3TC were added into the cells. Cell control without medicine was set at the same time. Cultural supernatants were collected at 9 d. HBsAg and HBeAg in cultural supernatants were tested by ELISA. The content of HBV-DNA was detected by TaqMan probe fluorescence quantitative PCR.</p><p><b>RESULT</b>TC0 and TC50 of PCC in HepG2 2. 2. 15 cell culture were 1 g . L-1 and >10 g . L-1, respectively, suggesting low toxicity in cells. IC50 of PCC in HepG2 2. 2. 15 cells HBsAg and HBeAg were 0. 501, 0. 401 g. L-1, with SI being >19.96 and >24. 94, respectively. PCC could effectively inhibit the secretion of HBsAg and HBeAg, and have a better effect on HBeAg than on HBsAg. PCC had a significant inhibitory effect on HBV-DNA in HepG2 2. 2. 15 cells at concentrations of 0. 1, 1 g . L-1 P <0.05).</p><p><b>CONCLUSION</b>PCC has the effect against HBV activity in vitro to some extent, with low toxicity, thereby having a good prospect for application.</p>


Subject(s)
Animals , Antiviral Agents , Pharmacology , DNA, Viral , Metabolism , Hep G2 Cells , Hepatitis B Surface Antigens , Metabolism , Hepatitis B e Antigens , Metabolism , Hepatitis B virus , Metabolism , Humans , Polysaccharides , Pharmacology , Snails , Chemistry
14.
Article in Chinese | WPRIM | ID: wpr-814718

ABSTRACT

OBJECTIVE@#To evaluate the change of plasma level of retinol binding protein 4 (RBP4) in patients with coronary heart disease, and to explore the effect of hyperinsulinemia.@*METHODS@#This study was carried out at Xiangya Hospital of Central South University, China, from September 2009 to May 2010. Thirty patients with coronary artery disease (the CAD group) were confirmed by coronary angiography, 29 patients with CAD plus hyperinsulinemia (the CAD+HIns group), and 30 healthy subjects were enrolled as controls (the control group). The peripheral blood sample from the anticubital vein was collected aseptically in all the subjects to measure the RBP4 by enzyme linked immunosorbent-assay (ELISA). The height, weight, body mass index (BMI) the waist-to-hip ratio (WHR), the blood pressure, the fasting plasma glucose (FPG), the fasting insulin (Fins), the 2-hour postprandial inslulin (2hPIns), and the homeostasis model assessment-insulin resistance index (HOMA-IR) was measured. The lipids, high sensitivity C reactive protein (hsCRP), uric acid(UA), free fatty acids (FFA) were all examined.@*RESULTS@#The level of plasma RBP4 in the CAD+HIns group was higher than that in the CAD group and the control group (both P0.05). Correlation analysis showed that the plasma RBP4 level was significantly correlated with BMI, FPG, FIns, 2hPIns, HOMA-IR, TG, HDL-C, UA, and hsCRP (r=0.259, 0.331, 0.582, 0.452, 0.600, 0.236, -0.290, 0.243, 0.231, respectively; all P>0.05). Multiple regression analysis showed that BMI, 2hPIns, and HOMA-IR were the independent factors related to RBP4.@*CONCLUSION@#The plasma level of RBP4 does not increase in the CAD group, but it is high in the CAD +HIns group. RBP4 level is related to BMI, lipids, UA, and other cardiovascular risk factors. BMI, 2hPIns, and HOMA-IR are the independent factors associated with RBP4.


Subject(s)
Adult , Aged , Case-Control Studies , Coronary Disease , Blood , Female , Humans , Hyperinsulinism , Blood , Insulin Resistance , Physiology , Lipids , Blood , Male , Middle Aged , Retinol-Binding Proteins, Plasma , Metabolism , Uric Acid , Blood
15.
Article in Chinese | WPRIM | ID: wpr-814700

ABSTRACT

OBJECTIVE@#To examine the plasma adiponectin concentration in coronary heart disease (CHD) patients combined with abnormal glucose metabolism, and to explore the clinical significance of adiponectin.@*METHODS@#Eighty-seven hospitalized CHD patients confirmed by coronary angiography from August 2009 to April 2010 at Xiangya Hospital were enrolled and divided into 3 groups according to their glucose metabolic state: 31 patients were selected as a simple CHD group, 28 were selected as a CHD combined with impaired glucose tolerance group (CHD+IGT group), and the other 28 as a CHD combined with diabetes mellitus group (CHD+DM group). The 31 healthy subjects who got health checkup at the same time were enrolled as a normal control group (NC group). Plasma adiponectin was measured by enzyme linked immunosorbent assay. The height, weight,waistline and blood pressure of all the subjects were checked, and the fasting blood glucose (FBG), insulin, lipids, high-sensitivity C-reactive protein (hs-CRP), free fatty acids (FFA), the liver function and the renal function were checked as well. The body mass index and the homeostasis model were assessed for insulin resistance.@*RESULTS@#1) Plasma adiponectin in the CHD group, the CHD+IGT group, and the CHD+DM group was all lower than that in the NC group (P<0.05); 2) Compared with the CHD group, the plasma adiponectin in the CHD+DM group was the lowest, followed by the CHD+IGT group, and there was significant difference in the 3 groups (P<0.05); 3) Plasma adiponectin level was positively related with the high density lipoprotein cholesterol-C (HDL-C) (r=0.483, P<0.01), while it was negatively related with the hs-CRP and Gensini score (r=-0.489, P<0.05;r=-0.252, P<0.05).@*CONCLUSION@#Plasma adiponectin concentration is reduced in the CHD patients, and significantly reduced in CHD patients combined with abnormal glucose metabolism. Plasma adiponectin concentration decreases significantly with the severity of abnormal glucose metabolism. CHD and the abnormal glucose metabolism are important influence factors for plasma adiponectin. That plasma adiponectin level significantly decreases may be the superimposed results of CHD and abnormal glucose metabolism. Plasma adiponectin combined with HDL-C, hs-CRP and Gensini score may provide the reference in the judgement of the severity of CHD patients with abnormal glucose metabolism.


Subject(s)
Adiponectin , Blood , Aged , Coronary Disease , Blood , Metabolism , Diabetes Mellitus, Type 2 , Blood , Female , Humans , Insulin Resistance , Physiology , Male , Middle Aged
16.
Article in Chinese | WPRIM | ID: wpr-281666

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the inhibition of Streptococcus oligofermentans (So) on Streptococcus mutans (Sm) and the producibility of hydrogen peroxide by So under the influence of glucose concentration environment.</p><p><b>METHODS</b>The inhibition between So and Sm was observed by plating method under the different glucose concentration environment. The initial synthesis rates and production of hydrogen peroxide by So were determined under the different glucose concentration environment by 4-aminoantipyine-horseradish peroxidase method at A(510).</p><p><b>RESULTS</b>Under 0, 10 and 50 mmol/L glucose environment, the inhibition of So on Sm was evident. When both Sm and So were inoculated at the same time, the ratio of inhibition area by bacterial membrane area was 0.202 ± 0.005, 0.467 ± 0.025, 0.468 ± 0.028 under 0, 10, 50 mmol/L glucose environment. When So was cultivated first and then Sm applied, the ratio was 0.394 ± 0.004, 0.811 ± 0.075 and 0.816 ± 0.007 under 0, 10 and 50 mmol/L glucose environment respectively. The inhibition under 10 and 50 mmol/L glucose environment were more significant than that under non-glucose environment. There was no significant difference between these two glucose concentrations (P > 0.05). The initial synthesis rates of H2O2 by So under the 10 mmol/L [(23.573 ± 0.263) µmo×L(-1)×min(-1)] and 50 mmol/L [(23.337 ± 0.473) µmol×L(-1)×min(-1)] glucose were higher than without glucose[(10.513 ± 0.516) µmol×L(-1)×min(-1)], P < 0.05. H2O2 was not detected in 1000 mmol/L glucose. However, the production of H2O2 by So under 0 mmol/L glucose was higher than other glucose concentrations (P < 0.05).</p><p><b>CONCLUSIONS</b>The capability of the inhibition of So on Sm was affected by glucose environment and was much stronger under certain glucose concentrations (10, 50 mmol/L).</p>


Subject(s)
Antibiosis , Dose-Response Relationship, Drug , Glucose , Metabolism , Hydrogen Peroxide , Metabolism , Streptococcus , Metabolism , Physiology , Streptococcus mutans , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-246168

ABSTRACT

<p><b>OBJECTIVE</b>To investigate human enterovirus 71 (EV71) resistance to type I interferon induced antiviral effect.</p><p><b>METHODS</b>After type I interferons (alpha, beta) were incubated with HeLa cells, recombinant type I herpes simple virus (HSV-1) with green fluorescent protein (GFP) was inoculated onto the HeLa cells. HSV-1 proliferation was observed by GFP expression and PCR. After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon, proliferation of EV71 were detected by RT-PCR of 2A gene.</p><p><b>RESULTS</b>Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type I interferon (alpha, beta). However, EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR.</p><p><b>CONCLUSION</b>HeLa cell irritated by type I interferon (alpha, beta) produced antiviral substance that inhibits HSV-1 proliferation. EV71 resisted the antiviral substance induced by type I interferon and could significantly replicate in the HeLa cells.</p>


Subject(s)
Antiviral Agents , Pharmacology , Drug Resistance, Viral , Enterovirus A, Human , HeLa Cells , Humans , Interferon Type I , Pharmacology , Virus Replication
18.
Chinese Journal of Virology ; (6): 57-62, 2012.
Article in Chinese | WPRIM | ID: wpr-354771

ABSTRACT

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.


Subject(s)
Arboviruses , Genetics , Encephalitis Virus, Japanese , Genetics , Encephalitis Viruses , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sensitivity and Specificity
19.
Article in English | IMSEAR | ID: sea-136411

ABSTRACT

Background:Interleukin-17A (IL-17A) is a key inflammatory cytokine in many disorders, while the significance of IL-17A in nasal polyposis (NP) is still obscure. This study aimed to investigate the expression of IL-17A in nasal polyps from both atopic and nonatopic patients and its associations with clinical and histological features. Methods: In all, 30 patients with NP were included, and were grouped into atopic and nonatopic patients according to skin prick test (SPT). Disease severity was evaluated by symptom score, endoscopy score and CT score. Histological characteristics were assessed by eosinophilic infiltration, basement membrane (BM) thickness, epithelial damage, squamous metaplasia, and goblet cell hyperplasia. IL-17A expression in polyps was detected by ELISA and immunohistochemistry. Results: Endoscopy score and CT score were significantly higher in atopic NP patients than in nonatopic NP patients (P<0.05). IL-17A levels were significantly upregulated in both atopic (P<0.01) and nonatopic (P<0.05) patients versus controls. Furthermore, IL-17A levels were significantly higher in the atopic group versus nonatopic group. Significantly positive correlations were found between IL-17A levels and CT scores, eosinophilic infiltration and BM thicknesses. Conclusions: These results indicated that expression of IL-17A was significantly upregulated in NP patients and was more severe in atopic NP patients, suggesting that IL-17A may play an important role in the pathology of NP and atopy may contribute to NP by stimulating the production of IL-17A.

20.
Article in Chinese | WPRIM | ID: wpr-298597

ABSTRACT

Allergic rhinitis (AR),with an increasing uptrend of the prevalence in many developed and developing countries,is a global health problem that affects people of all ages and ethnic groups.However,data on the prevalence of self-reported AR in western China are rare.This study investigated the epidemiological features of self-reported AR in western China.In the cross-sectional,population-based study,a validated questionnaire survey on self-reported AR was carried out in 4 major cities in western China by multistage,stratified and cluster sampling,from January to December 2008.The total prevalence rate was 34.3%,with 32.3% (Chongqing),34.3% (Chengdu),37.9% (Urumqi),30.3% (Nanning),respectively.The prevalence presented to increase with age before 30 years old while decrease with age after 30 years old,and the highest prevalence was in 19-30 years group in Chongqing,Chengdu and Nanning which significantly showed “persistent and moderate-severe” type (P<0.0001); In Urumqi,there wasn't a significant increasing or decreasing trend of prevalence rate with age but with an “intermittent and mild”predominance (P<0.0001).There were no distinct sexual differences in prevalence rates in the 4 cities.The morbidity was positively related to monthly average temperature and sunshine (r=0.76645,P=0.0036; r=0.67303,P=0.0165),but negatively associated with relative humidity (r=-0.64391,P=0.0238) in Urumqi.Interestingly,the monthly morbidity was negatively associate with average temperature,sunshine and precipitation in Nanning (r=-0.81997,P=0.0011; r=-0.60787,P=0.0360; r=-0.59443,P=0.0415).Self-reported AR is becoming common in western China with a rapid development in recent years,affecting about three persons out of ten.The climatic factors may have an indirect impact on the prevalence rate through the effects on the local allergens.

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