1.Rapid detection of Salmonella by enzymatic recombinase amplification combined with lateral flow chromatography
NIE Yanni ; YAN Meiying ; SONG Yanyan
Journal of Preventive Medicine 2023;35(12):1102-1104
Objective:
To establish a rapid detection method for Salmonella based on the combination of enzymatic recombinase amplification (ERA) and lateral flow chromatography (LF), so as to provide technical support for the on-site detection of Salmonella.
Methods:
Specific ERA primers and probes were designed based on the highly conserved flagella gene fimY in Salmonella. The primers were screened using capillary electrophoresis, and the probes were designed according to the amplification range of the screened primers. The amplification temperature and time were optimized to establish the amplification method, and the product was detected using LF strips. A standard strain of Salmonella was used to verify the sensitivity, 10 other gut bacteria were used to to verify the specificity and sensitivity, and the nucleic acid of the actual Salmonella strains was amplified to verify the detectability.
Results:
After screening for Salmonella-specific primers using capillary electrophoresis, the minimum detection concentration was 5 copies/μL under the amplification temperature of 37 ℃ and reaction time of 20 minutes. This method had a positive amplification result for Salmonella nucleic acid, and the amplification results of 10 other gut bacteria were all negative, with good specificity.
Conclusion
This method provides a possibility for on-site point of care testing of Salmonella infection.
2.Dynamic Changes of Four Main Active Components in Wine-processed Cuscuta Chinensis with Different Processing Methods
Qiuli LI ; Yanfang LIU ; Mengxiao YAN ; Yanni SONG
China Pharmacist 2016;19(10):1882-1884
Objective:To study the dynamic changes of the main active components in wine-processed Cuscuta chinensis with dif-ferent rocessing methods. Methods: An HPLC method was adopted for the content determination of chlorogenic acids, hyperoside, quercetin and kaempferol in C. chinensis and wine-processed C. chinensis with different baking temperature, baking time and amount of yellow wine. The chromatographic conditions were as follows: the detection wavelength was 360 nm, and methanol-0. 1% phosphoric acid was used as the mobile phase with gradient elution. Results:The wine processing system could increase the content of quercetin and kaempferol, while decrease the content of chlorogenic acids and hyperin. Conclusion: Different processing methods have certain effects on the main active components, which provide basis for the further study on the processing mechanism and quality control of wine-processed C. chinensis.
3.Synthesis and structure-activity relationship of 13-hexylberberine analogues as CD36 antagonists.
Yinghong LI ; Li WANG ; Bin HONG ; Yanni XU ; Shuyi SI ; Jiandong JIANG ; Danqing SONG
Acta Pharmaceutica Sinica 2010;45(9):1128-33
Scavenger receptor CD36 could bind and endocytose oxLDL into macrophages which were then differentiated into foam cells that constitute the atherosclerotic lesion core, and was considered to be a potential target to treat atherosclerosis. In the establishment of the compound library of berberine (BBR, 1) analogues, we discovered that 13-hexylberberine (2) showed an antagonistic activity against CD36. Taking 2 as the lead compound, 21 derivatives were synthesized and their antagonistic activities were evaluated via an ELISA-like high-throughput screening (HTS) model. The primary structure-activity relationships were studied. It was indicated that the introduction of suitable groups at the 2- and 3-position of the aromatic ring A or at the 9-position of the aromatic ring D could enhance the activity. Among the 21 studied compounds, 7g bearing a benzyloxyl group at the 9-position provided a highest CD36 antagonistic activity with the IC50 value of 7.7 micromol L(-1). Besides, its antagonistic activity was further verified with Sf9 insect cell HTS model. So berberine analogues are a new family of CD36 receptor antagonists and worthy to be studied further.
4.Establishment and application of multiplex touchdown PCR for detection of extended-spectrum β-lactamases-producing Enterobacteriaceae and me-thicillin-resistant Staphylococcus aureus in bloodstream infection
Yanni BI ; Zhengeng SUI ; Yu SONG ; Yemin QU ; Shuqing MA ; Mei SUN ; Haizhu LIU
Chinese Journal of Infection Control 2015;(10):663-667
Objective To develop a multiplex touchdown PCR for simultaneous detection of extended-spectrum β-lactamases (ESBLs )-producing Enterobacteriaceae and methicillin-resistant Staphylococcus aureus (MRSA ). Methods Blood culture positive specimens from 102 hospitalized patients were collected between March 2013 and September 2014,four pairs of specific primers were designed based on SHV,TEM,and OXA genes of ESBLs-pro-ducing Enterobacteriaceae and MecA gene of MRSA,drug-resistant genes were amplified with single touchdown PCR and multiplex touchdown PCR, the results were compared with Kirby-Bauer disk diffusion method. Results Each single PCR amplified a specific band,four drug-resistant genes were also detected by multiplex touchdown PCR;the lower detection limits of multiplex touchdown PCR for DNA of MecA,SHV,TEM,and OXA were 4.37 ng,2.19 ng,4.53 ng,and 3.59 ng,respectively.Compared with Kirby-Bauer disk diffusion method, the overall sensitivity and specificity of multiplex touchdown PCR were 100.00% and 88.24% respectively,for ES-BLs were 100.00% and 87.23% respectively,for MRSA were both 100.00%.Conclusion A higher sensitivity and specificity multiple touchdown PCR assay has been developed,and it can be used in the rapid diagnosis and epidemi-ology investigation of bloodstream infection caused by ESBLs-producing Enterobacteriaceae and MRSA,and is help-ful for guiding antimicrobial use in clinic.
5.Related research of male breast cancer and CAG repeat polymorphism of AR gene
Jialin CUI ; Rui HUANG ; Yongdong JIANG ; Jiguang HAN ; Ming NIU ; Wei WEI ; Wei ZHENG ; Yanni SONG
Practical Oncology Journal 2015;(6):492-496
Objectiv e To investigate the correlation between ( CAG) n repeat polymorphism of androgen receptor(AR)geneandmalebreastcancer.Methods 40casesofmalebreastcancerand40controlswerecol-lected.DNA was extracted from peripheral blood and the AR gene CAG coding exon sequences for PCR amplifica -tion,sequencing and calculated the number of CAG repeats frquency .χ2 test and Logistic regression analysis were used assess the AR gene CAG repeat length frequency affect the number of male breast cancer risk .Results There was statistically significant difference in male breast cancer cases and controls the number of CAG repeat length frequency.Man for whom the(CAG)n≥22 repeat sequence had 3.52 times risk of male breast compared (CAG)n≤22(OR=3.52,P=0.036).Conclusion AR gene CAG repeat length is a predictor of the frequency of male breast cancer risk .Longer CAG repeats can increase the risk of male breast cancer .
6.Negative expression of RECK indicates unfavorable clinical outcome for breast cancer
Shaoqiang CHENG ; Yang LIU ; Xiaoshuan ILANG ; Da PANG ; Guoqiang ZHANG ; Jinsong WANG ; Yanni SONG
Practical Oncology Journal 2014;(1):12-18
Objecive To explore the significance of RECK expression in breast cancer .Methods Im-munohistochemical staining was used to analyze RECK expression levels in patients with breast cancer .We com-pared these data with the clinicopathological features of these patients .Rseults Breast cancer patients with nega-tive RECK expression had significantly lower DFS and 5-year survival rates than patients with positive RECK expression.In addition,for node-negative breast cancer ,negative RECK expression indicated markedly unfavor -able survival rate than positive arm .Multivariate analysis further confirmed that RECK expression was an inde -pendent prognostic factor for patients with breast cancer .Conclusion The loss of RECK expression indicates un-favorable survival rate for patients with breast cancer .RECK expression is a new ,important risk factor for recur-rence in breast cancer .
7.The Effects of Taurine Postconditioning in Pancreatic Injury Following Limb Ischemia Reperfusion in Rats
Yanni ZHENG ; Jing WU ; Yangyang HOU ; Jing ZHAO ; Xudong SONG ; Xiuli MEN
Tianjin Medical Journal 2013;(12):1188-1190
Objective To observe changes of the pancreas after limb ischemia reperfusion, and the protective effect of taurine (Tau) postconditioning. Methods Twenty-four male SD rats were randomly divided into control group (Control), limb ischemia reperfusion (LIR) group, and taurine postconditioning (Tau) group (n=8 for each group). Plasma contents of xanthine oxidase (XOD), malondialdehyde (MDA), reactive oxygen species (ROS), superoxide dismutase (SOD), pancreatic amylase (AMS), pancreatic lipase (LPS) and trypsin were detected in three groups. Morphological changes of the pancreas were observed using optical microscopy. The expressions of anti-C/enhancer-binding protein homologous protein (CHOP) in pancreas were analyzed by immunohistochemistry assay. TUNEL staining was performed to estimate the apoptosis of pancre-atic cells. Results Compared with the control group and Tau group, plasma contents of MDA, XOD, ROS, AMS, LPS, and trypsin were significantly increased in LIR group, but the level of SOD was significantly lower in LIR group ( P<0.05). HE staining showed that acinar structure was disrupted , pancreatic lobule gap widened, stromal vascular dilatation and conges-tion were observed in LIR group. The perivascular infiltration of inflammatory cells appeared, islet cells were damaged with irregular islet. The immunohistochemical analysis showed the increased expression of CHOP in pancreas in LIR group than that of Tau group. And the pancreatic apoptosis was enhanced in LIR group detected by TUNEL staining. Conclusion Tau-rine postconditioning can ameliorate pancreatic injury following limb ischemia reperfusion, which may be related to the inhi-bition of oxidative stress, down-regulation of CHOP expression, thereby reducing endoplasmic reticulum stress-induced apoptosis.
8.Optimization of Water Extraction Process for Glycyrrhiza by Central Composite Design/Response Surface Methodology
Chuan GU ; Jing WU ; Yanni LIANG ; Zhongxing SONG ; Hongbo LIU ; Shijun LIU
China Pharmacist 2015;(8):1253-1255
To optimize the water extraction process for glycyrrhiza. Methods: HPLC was used to determine the con-tents of glycyrrhizic acid and liquiritin. The comprehensive index included the contents of glycyrrhizic acid and liquiritin and the yield of dry extract. The water amount and the extraction time were selected as the independent variables, and the comprehensive index was set as the dependent variable. Design-expert 8. 06 software was used to fit multivariate linear or quadratic multinomial models for the experimental values. Response surface methodology was used to optimize the water extraction process. The prediction was carried out through comparing the observed and predicted values. Results:The regression coefficient of binomial fitting complex model was as high as 0. 979 7. The optimum conditions of extraction process were as follows:12-fold amount of water, extracting 3 times with 90 min for each time. The deviation between the observed and predicted values was -1. 72%. Conclusion: Central composite design-response surface methodology is convenient and highly predictive in optimizing the water extraction process for glycyrrhiza, which can be applied in the further membrane separation and purification.
9.Application value of immunoglobulin G4 detection in unexplained liver injuries
Yue ZHANG ; Lu ZHOU ; Guohui JIAO ; Yanni LI ; Yujie ZHANG ; Wenjing SONG ; Jie ZHANG ; Bangmao WANG
Chinese Journal of Digestion 2017;37(8):530-536
Objective To investigate the expression level of immunoglobulin G (IgG)4 cells in patients previously diagnosed as unexplained liver injuries,and to analyze whether these patients could be diagnosed as immunoglobulin G4-related disease (IgG4-RD).The clinical characteristics of patients with IgG4 positive cells infiltration in liver tissues who did not met IgG4-RD diagnosis criteria were also analyzed.Methods From August 2010 to July 2016,33 patients with unexplained liver injuries even after liver biopsy were enrolled.Immunostaining of IgG and IgG4 in liver tissue slices was performed.Whether the patients met the IgG4-RD diagnosis criteria was analyzed.Also,the relation between IgG4 positive cell infiltration in liver tissues and serum parameters (alanine aminotransferase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),γ-glutamyl transpeptadase (GGT),IgG),the classification of liver inflammation and stage of liver fibrosis were analyzed.Chi-square,Fisher exact probability method,t test and Wilcoxon rank and inspection were used for statistical analysis.Results Among 33 patients with unexplained liver injuries,average age of onset of the disease was (50.48 ± 14.52) years old.Ten patients (30.3 %) presented with different degree of IgG4 positive cells infiltration in liver.In one patient,the IgG4 positive cell number in liver tissue was over 10/high-powered field (HPF),IgG4+ / IgG+ ratio was over 40%,the serum IgG4 increased and low density nodules in the left hepatic lobe level,which met the diagnostic criteria of IgG4-RD.Serum parameters of liver function of this patient were as follows:ALT 948 U/L,AST 658 U/L,ALP 176 U/L,GGT 197 U/L,TBil 21.5 μmol/L,DBil 10.0 μmol/L.According to whether there were IgG4 positive cells infiltration in liver tissues,33 patients were divided into IgG4 positive cells infiltration group (IgG4 positive group,10 cases) and no IgG4 positive cells infiltration group (IgG4 negative group,23 cases).The medium levels of ALT,AST,ALP and GGT in the IgG4 positive group and the IgG4 negative group were 26.0 U/L vs 31.0 U/L,28.5 U/L vs 32.0 U/L,114.0 U/L vs 77.0 U/L,65.0 U/L vs 36.0 U/L,respectively and there were no statistically significant differences (Z=-0.098,-0.216,-0.862,-0.705;all P>0.05).The serum IgG level of the IgG4 positive group was higher than that of the IgG4 negative group (17.15 g/L vs 13.65 g/L),and the difference was statistically significant (Z=-1.776,P =0.035).Conclusions IgG4-RD may be the reason of liver injury.There are no significant differences in clinical charateristics,serum parameters and inflammation degree in liver tissue between patients with or without IgG4 positive cells infiltration.
10.Clinical effect of phase Ⅰ cardiac rehabilitation exercise on patients with acute myocardial infarction after percutaneous transluminal coronary intervention
Kaili FAN ; Huiyun YANG ; Yanni WU ; Yafan SONG ; Tian ZHOU ; Rong WANG
Chinese Journal of Practical Nursing 2021;37(18):1394-1401
Objective:To explore the clinical effect of phase Ⅰ cardiac rehabilitation exercise on patients with acute myocardial infarction after percutaneous transluminal coronary intervention (PCI).Methods:Convenient sampling method was adopted, totally 84 acute myocardial infarction patients after PCI was randomized into an observation group and control group. Both groups accepted general nursing care. The observation group also accepted the phase Ⅰ cardiac rehabilitation exercise. Cardiac ultrasonic, the 6-minute walk test were used to evaluate the patients′ cardiac function and exercise tolerance, the SF-12 were used to evaluate the quality of life.Results:After repeated measurement ANOVA, before the intervention, there was no significant difference in cardiac function and quality of life between the two groups ( P>0.05); before discharge, the 6-minute walk distance of the observation group was longer than that of the control group, and the difference was significant( F value was 5.279, P=0.024). At 1 month after discharge, there were significant differences in the LVEF( F value was 8.119, P=0.006) and 6-minute walking distance( F value was 9.829, P=0.002) between the two groups ( P<0.05), analysis of the six items of SF-12 including general health( F value was 6.905, P=0.010), physical functioning( F value was 10.595, P=0.002), role physical( F value was 11.168, P=0.001), bodily pain( F value was 12.548, P=0.001), mental health( F value was 7.362, P=0.008) and vitality( F value was 13.692, P<0.001) having shown significant differences between the two groups. At 3 months after discharge, there were significant differences in the LVEF( F value was 11.156, P=0.001), 6-minute walk distance( F value was 16.554, P<0.001)and quality of life in all dimensions between the two groups ( P<0.05). Conclusion:Phase Ⅰ cardiac rehabilitation exercise can improve cardiac function and the quality of life in patients with acute myocardial infarction undergoing PCI, and enhance the exercise tolerance.