By introducing an electro-withdrawing antipyrine group, N-(p-toluenesulfonyl)-N-(4-antipyrine)-10-methylacridinium-9-carboxamide triflate was prepared. The UV, FL and CL properties of the target compound and of its precursor were investigated by comparing with those of the model compound N-(p-toluenesulfonyl)-N-phenyl-10-methylacridinium-9-carboxamide triflate and the corresponding precursor respectively. The results show that acridine sulfonamide with a heterocyclic antipyrine group exhibits blue shift of both UV absorption and of maximum excitation wavelength(λex) and emission wavelength(λem) in FL spectra, comparing with the corresponding model compound. The λex of the final target and its precursor are 268 and 274 nm, respectively; and the λem are 321 and 327 nm, respectively, while λex of the model compound and its unmethylated precursor are 365 and 359 nm, respectively; and the λem are 504 and 440 nm, respectively. Moreover, the chemiluminescence of the final target compound triggered by H2O2 could finish within 1.1 s; and the quantum yield is similar to that of the model compound, being 5.6 times high as that of luminol.

Molecularly imprinted polymers for dimethoate recognition were synthesized by the precipitation polymerization technique using methyl methacrylate (MMA) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker. The morphology, adsorption and recognition properties were investigated by scanning electron microscopy (SEM), static adsorption test, and competitive adsorption test. To obtain the best selectivity and binding performance, the synthesis and adsorption conditions of MIPs were optimized through single factor experiments. Under the optimized conditions, the resultant polymers exhibited uniform size, satisfactory binding capacity and significant selectivity. Furthermore, the imprinted polymers were successfully applied as a specific solid-phase extractants combined with high performance liquid chromatography (HPLC) for determination of dimethoate residues in the cucumber samples. The average recoveries of three spiked samples ranged from 78.5% to 87.9% with the relative standard deviations (RSDs) less than 4.4% and the limit of detection (LOD) obtained for dimethoate as low as 2.3μg/mL.

Objective To clone silent information regulator 1 (SIRT1) gene full-length cDNA,construct recombinant eukaryotic expression vector containing SIRT1 gene and its mutant T200I,E420K,so as to lay the foundation for further research of SIRT1 gene function.Methods RT-PCR amplified SIRT1 gene full-length cDNA.PCR products were cloned into the eukaryotic expression vector pcDNA3.1 (+) through double digestion and pcDNA3.1(+)-SIRT1 recombinant plasmid was obtained.Meanwhile,site-directed mutagenesis was applied to build its mutant pcDNA3.1 (+)-T200I and pcDNA3.1 (+)-E420K expression vector.Recombinant plasmid was identified by enzyme digestion and DNA sequencing and the recombinant eukaryotic expression vector of success was screened out.Results SIRT1 gene full-length cDNA was successfully cloned,and pcDNA3.1 (+)-SIRT1 eukaryotic expression vector and its mutant were also successfully constructed.Positive recombinant plasmid sequencing was compared after enzyme digestion,and it was completely consistent with the expected sequence.Transfected 293T cell line was established and HIS tagged SIRT1 protein was expressed.Conclusions We successfully constructed pcDNA3.1 (+)-SIRT1 and its mutant pcDNA3.1 (+)-T200I,pcDNA3.1 (+)-E420K eukaryotic expression vector,which may provide genetic material for biological function study of SIRT1 gene and its mutant T200I,E420K.

Objective To amplify the recombinant adenovirus vector carrying rat angiotensin Ⅱ type 2 receptor (AT2R) gene using human embryonic kidney (HEK) 293A cell lines and to construct a pancreatic islet βcell model overexpressing AT2R by transfecting the adenovirus vector into rat insulinoma (INS-1) cell lines.Methods Recombinant adenovirus vector Ad-G-AT2R-EGFP and control vector Ad-CMV-EGFP were amplified with HEK 293A cells and the titer of the adenovirus was detected .After both adenovirus vectors were transfected into INS-1 cells,AT2R and angiotensin Ⅱtype 1 receptor(AT1R) gene expressions were tested using real-time PCR, Western blotting, immunofluorescence staining and confocal laser-scanning microscopy .Results The titer of amplified Ad-G-AT2R-EGFP and Ad-CMV-EGFP was re-spectively 9 ×109 pfu/ml and 8 ×109 pfu/ml.Transfection of Ad-G-AT2R-EGFP into INS-1 cells induced an increase in AT2R mRNA expression in a dose-dependent manner , and significantly increased AT2R mRNA and protein expression compared with Ad-CMV-EGFP-or mock-transfection.Conclusion The recombinant adenoviral vector carrying AT2R gene is successfully amplified and an INS-1 cell model overexpressing AT2R is constructed by transient transfection , which can contribute to further study of the role of AT2R in pancreatic islet βcells.

OBJECTIVE:To improve the dispersion stability in water of Tussilago farfara powder,and to improve compliance of Xiao’er feike granules. METHODS:The effects of 4 kinds of dispersion stabilizer (sodium hexametaphosphate, dextrin, PEG4000 and lecithin) on dispersion stability of suspension in water were investigated during the grinding of T. farfara using rate of absorbance change(β)and Zeta potential as index;IR spectrum of samples were characterized. Using original formulation with-out dispersion stabilizer as control,the dispersion stability of new formulation granules in water were analyzed comparatively after adding dispersion stabilizer. RESULTS:Among 4 kinds of dispersion stabilizer,β of sample prepared by sodium hexametaphos-phate was the lowest,while Zeta potential of it was the highest;compared with original T. farfara,β of T. farfara grinded with 2.5% sodium hexametaphosphate decreased by 16.8%,and Zeta potential absolute value increased by 29.4%;no new peak was found in IR spectrum. Compared with control granules,granules suspension prepared by new formulation had lower β and higher Zeta potential absolute value (P<0.01);particle size was 30 μm and no large particle aggregation was found;β was less than 5.0% within 20 s sedimentation. CONCLUSIONS:During the preparation of Xiao’er feike granules,the application of sodium hexametaphosphate in the grinding of T. farfara powder can improve the dispersion stability of granules in water and the compliance of the preparation.

AIM: To investigate the effects of angiotensin-Ⅱ (Ang-Ⅱ) on the proliferation and collagen synthesis in rat cardiac fibroblasts and the expression of Ang-Ⅱ receptor on fibroblasts. METHODS: [3H]-TdR and [3H]-prolin at different concentrations were incubated with the confluent fibroblasts of newborn rats stimulated by Ang-Ⅱ. Receptor of Ang-Ⅱ on fibroblasts and intracellular free calcium ions were examined by autoradiograms and fluorescence technique. RESULTS: In the presence of Ang-Ⅱstimulation, the increase in incorporation of [3H]-TdR and [3H]-prolin was much higher than that of control in a dose-dependent manner. Autoradiograms showed that Ang-Ⅱ was uniformly distributed over the membrane of the fibroblasts. The silver grains on fibroblasts were obviously decreased after adding unlabeled Ang-Ⅱ by the autoradiograms. The concentration of free calcium ions was increased in fibroblast. CONCLUSION: These findings suggest that Ang-Ⅱ promotes fibroblast proliferation and the syntheses of collagen protein, in which Ang-Ⅱ binds to the membrane receptor of fibroblasts to activate the signal system in cells, such as intracellular free calcium ions. [

Objective To investigate short-term effect of percutaneous discectomy combined with radiofrequency ablation with Disc-FXon contained lumbar disc herniation. Methods 36 patients were reviewed and followed up with Japanese Orthopaedic Association score(JOA score), the Visual Analogue Score (VAS) and Oswestry score for 12 months. Results The scores of JOA score, VAS and Oswestry improvedsignificantly (P<0.01) after operation. Conclusion Percutaneous discectomy combined with radiofrequency ablation with Disc-FXis effective on contained lumbar disc herniation.

Objective To investigate the relationship between CYP 2C19 gene polymorphisms and clopidogrel efficacy in coronary heart disease patients after percutaneous coronary intervention (PCI).Methods From January 2016 to December 2017,62 patients with acute coronary syndromes and treated with PCI in Guizhou Aerospace Hospital were recruited, CYP2C19 genotype, ADP －induced platelet aggregation rate and myocardial enzymes and other indicators were detected before operation .The myocardial enzymes were measured 24 hours after PCI.According to different metabolic types,the patients were grouped,the above indicators were compared.Results The CYP2C19*1/*1 was 37.10%,CYP2C19*1/*2 was 35.48%,CYP2C19*1/*3 was 11.29%,CYP2C19*2/*2 was 12.90%,CYP2C19*2/*3 was 3.23% and CYP2C19*3/*3 was 0.00%.The LDH,AST,CK,CK－MB and α－HBDH in the PCI patients after operation were significantly higher than those before operation (t＝0.019,0.040, 0.044,0.022,0.014,all P＜0.05).But ADP induced platelet aggregation rate and myocardial enzymes and other indicators among fast metabolism group,intermediate metabolic group and slow metabolic group had no statistically significant differences (all P ＞0.05).Conclusion CYP2C19 mutation frequency in the Chinese population is relatively large,the sample size of this study is less ,the relationship between clopidogrel resistance and the specific genotype can not be obtained ,it need to increase the sample size and comprehensive multi －factor consideration .

Objective To comparatively analyze the clinical characteristics of thyrotropin-secreting pituitary adenomas (TSH-omas).Methods The clinical features, laboratory variables, imaging and pathological Results were retrospectively compared and analyzed of 26 cases with TSH-omas admitted in Chinese PLA General Hospital from Feb. 2006 to Oct. 2016 and 20 cases with TSH-omas admitted in Shanghai Huashan Hospital from Apr. 2006 to Apr. 2013.Results The female ratio was slightly higher in patients of Chinese PLA General Hospital than in Huashan Hospital [(57.7%(15/26)vs. 45.0%(9/20)], while the mean age was similar [39.5±14.1(18-67 years)vs. 40.0±14.5(17-74 years)]. The most common chief complaint was thyrotoxicosis [73.1%(19/26)vs. 55.0%(11/20)], and mild-to-moderate goiter was the most common symptom. The mean serum TSH levels in Chinese PLA General Hospital and in Huashan Hospital were 5.06(2.97-6.27)mU/L and 6.16(3.76-10.91)mU/L respectively, and patients with normal serum TSH levels were more common in Chinese PLA General Hospital than in Huashan Hospital [57.7%(15/26)vs. 40.0%(8/20)]. Microadenoma was more common in Chinese PLA General Hospital than in Huashan Hospital [34.62%(9/26)vs. 20.0%(4/20)], while macroadenoma was more common in Huashan Hospital than in Chinese PLA General Hospital [20.0%(4/20)vs. 7.7%(2/26)]. Microadenoma was more common in female patients of the both groups [66.7%(6/9)vs. 75.0%(3/4)], while macroadenoma was all found in male patients. Tumor invasion of surrounding tissue and structure was often found in macroadenoma. In terms of octreotide inhibition test, the range of 24h TSH inhibition rate was roughly the same in the two groups (37.4%-91.8% and 46.5%-94.1%, respectively). Mixed adenoma was rare among all the pathologically confirmed cases. In Chinese PLA General Hospitals, TSH immunoreactive negative neoplastic cells were found in 3 of 8 cases, and octreotide scanning showed negative in 2 of 12 cases.Conclusions Patients with TSH-omas in the two hospitals show similarities but also some significant differences in the clinical features. Overall, the domestic patients with TSH-omas are diagnosed without gender difference according to the reports in China. The mean age at diagnosis is significantly younger than that in foreign data. Microadenoma is more common in females, while macroadenoma is more common in males. Serum TSH levels can be normal in patients with TSH-omas. Immunostaining and/or octreotide scanning for TSH can be negative.

Objective: To investigate the infectious status, gene type transition and epidemiological features of rotavirus A isolated from infants and children (<59 months-of-age) in sentinel hospitals from 2008 to 2015 in Henan province, China. Methods: In total, 2 541 stool samples (each 3- 5 ml) were collected from infants and children aged less than five years in two sentinel hospitals and group A rotavirus was detected by a double antibody sandwich ELISA. Viral RNA was extracted from positive samples and G/P gene typing was performed using a two-step nested multiplex RT-PCR. Epidemiological information (including demographic information such as age, sex and clinical symptoms) was also collected from the patients and analyzed. Results: Group A rotavirus was detected in 30.9% (785/2 541) of diarrhea samples from children. The detection rate was higher in October (54.8%, 345/629) and lower in July (5%, 5/101) each year from 2008 to 2015. The group A rotavirus infection rate was higher in boys (30.6%, 451/1 476) than in girls (31.4%, 334/1 065) (χ²=0.18, P=0.664). Infection mainly occurred in 4-12 months old patients (61.3%, 481/785) (χ²=196.69, P<0.001), and the infection rate was lower in cities (26%, 258/992) compared with rural areas (34.0%, 527/1 549) (χ²=18.19, P<0.001). G typing of 785 strains of group A rotavirus revealed the following types: G1 (13.5%, 106 strains), G2 (11.1%, 87 strains), G3 (29.7%, 233 strains), and G9 (57.5%, 451 strains); P typing revealed the predominance of P[4] (11.3%, 89 strains) and P[8] (84.7%, 665 strains); gene type combinations comprised mainly G9P[8], G2P[4], G3P[8], G1P[8], respectively accounted for 52.9% (415), 9.7% (76), 17.3% (136), 11.3% (89). Gene type combinations G1 [8] and G3P[8] have been decreasing in prevalence since 2008 and G9P[8] has become the dominant gene type of group A rotavirus in Henan province. Among the group A rotavirus infection samples, the male:female infection ratio was 1.4∶1 (451/334), with no significant difference in the infection rate (χ²=0.18, P=0.664); the infection rate was higher in 4- 12 months old patients (61.3%, 481/785), with a significant difference detected between age groups (χ²=196.69, P<0.001). The rate of detection was lower in cities (26.0%, 258/992) than in rural areas (34.0%, 527/1 549) (χ²=18.19, P<0.001). Clinical analysis revealed a body temperature of below 37 degrees in 75.7% of positive cases (594 patients), 37.0- 37.5 degrees in 17.2% of cases (135 patients), 37.6-38.0 degrees in 2.0% of cases (16 patients), and above 38 degrees in 5.1% of cases (40 patients), with most cases showing no fever or a mild fever. The frequency of episodes of diarrhea among the patients was 0- 3 times (21.1%, 166 cases), 4- 6 times (65.6%, 515 cases), 7- 9 times (8.0%, 63 cases), or 10- 15 times (5.2%, 41 cases), mainly showing mild and moderate diarrhea. Vomiting also varied in frequency among the patients from no vomiting (86.9%, 682 cases), 1-2 times (11.8%, 92 cases), 3 times (6.0%, 47 cases), and more than 3 times (0.4%, 3 cases). The occurrence of dehydration varied from no dehydration (86.9%, 682 cases), mild dehydration of 1%- 5% (12.1%, 95 cases), to severe dehydration of ≥5% (1.0%, 8 cases). Conclusion A higher infection rate of group A rotavirus was detected in children younger than five years of age with acute diarrhea in sentinel hospitals in Henan province, including part-mixed infection cases. A predominance of cases was detected in the autumn, and secondly the spring of each year. Gene type G9P[8] was most frequently isolated. The majority of patients displayed no fever, vomiting or dehydration. The cases with clinical symptoms of fever, diarrhea, vomiting and dehydration often showed mild disease.