For the purpose of expanding the analysis scope of medicines,a RPHPLC assay procedure has been established for quantitative analysis of indomethacin in human plasma.Analytical column was YWG C18(?4 6 mm?200mm).Mobile phase was methanol water acetic acid solution(67∶33∶0 1)(v/v) and wavelength of detector was 254nm.The linear relationship,precision,method of extraction and recovery were comparatively investigated by standard blank human plasma spiked with indomethacin.Indomethacin leved in the blood of the healthy volunteers was detected by using this method.The linear range of the method was 0 1～5 0?g?ml -1 .The calibration curve was linear (?=0 9995). The detection limit was 0 02?g?ml -1 (S/N≥3) and the recovery of indomethacin in human blood was between 97 5%～104 2%. Intra and inter day prccision of the mothod were(1 1?0 2)%(n=4) and(2 7?0 6)%(n=4)respectively.The CV% were no more than 3 0% (n=4).The method shows a high sensitivity,precision,fast and excellent selectivity.Thus it is suitable for investigation of the indomethacin in human blood and its toxicological analysis as well as the pharmacokinetic study.

Altitude ; Animals ; Astrocytes ; cytology ; Brain ; cytology ; Hypoxia ; Male ; Microglia ; cytology ; Rats ; Rats, Wistar

The CrdS protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to explore the regulatory mechanism in the acidic adaptation of Helicobacter pylori (H. pylori). The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS protein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm-T-hp1364 and the prokaryotic expression plasmid pQE30-hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XL1 blue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.
Bacterial Proteins ; biosynthesis ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Helicobacter pylori ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

Drug-Eluting Stents ; adverse effects ; Humans ; Male ; Middle Aged ; Prosthesis Failure ; Thrombosis ; etiology

The aim of this paper is to report the synthesis of the mPEG-PCL-g-PEI copolymers as small interfering RNA (siRNA) delivery vector, and exploration of the siRNA delivery potential of mPEG-PCL-g-PEI in vitro. The diblock copolymers mPEG-PCL-OH was prepared through the ring-opening polymerization. Then, the hydroxyl terminal (-OH) of mPEG-PCL-OH was chemically converted into the carboxy (-COOH) and N-hydroxysuccinimide (NHS) in turn to prepare mPEG-PCL-NHS. The branched PEI was reacted with mPEG-PCL-NHS to synthesize the ternary copolymers mPEG-PCL-g-PEI. The structure of mPEG-PCL-g-PEI copolymers was characterized with Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and gel permeation chromatography (GPC). The mPEG-PCL-g-PEI/siRNA nanoparticles were prepared by complex coacervation, and the nanoparticles size and zeta potential were determined, separately. The cytotoxicities of mPEG-PCL-g-PEI/siRNA nanoparticles and PEI/siRNA nanoparticles were compared through cells MTT assays in vitro. The inhibition efficiencies of firefly luciferase gene expression by mPEG-PCL-g-PEI/ siRNA nanoparticle at various N/P ratios were investigated through cell transfection in vitro. The experimental results suggested that the ternary (mPEG5k-PCL(1.2k))1.4-g-PEI(10k) copolymers were successfully synthesized. (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k) could condense siRNA into nanoparticles (50-200 nm) with positive zeta potential. MTT assay results showed that the cytotoxicity of (mPEG(5k)-PCL(1.2k))1.4-g-PEI(10k)/siRNA nanoparticles was significantly lower than that of PEI(10k)/siRNA nanoparticles (P < 0.05). The expression of firefly luciferase gene could be significantly down-regulated at a range of N/P ratio from 50 to 150 (P < 0.01), and maximally inhibited at the N/P ratio of 125. The mPEG-PCL-g-PEI polymers could delivery siRNA into cells to inhibit the expression of target gene with very low cytotoxicity, which suggested that mPEG-PCL-g-PEI could serve as a new type of siRNA delivery vector.

Objective:To evaluate the correlation between the polysomnographic findings and the degree of obstruction caused by adenoid and tonsillar hypertrophy in children with clinical history of apnea. Method: Retrospectively studied the children who were diagnozed clinically of, obstructive sleep apnea hypopnea syndrome(OSAHS) and underwented polysomnograph and endoscopy. Patients were divided nto OSAHS and non-OSAHS group according to polysomnographic findings. Result: Ninty-four children were involved in the study population, and 63 children of them were male. The mean age of the children at the time of inclusion in the study was 6.7 years. 36 children(38.3%) diagnosed OSAHS clinically had normal polysomnographic findings. No differences were found between children with PSG-documented OSAHS and others. Tonsillar and/or adenoid hypertrophy were not correlated to more severe apnea among enrolled children. Conclusion-There was no significant correlation between polysomnographic and clinical findings in children with OSAHS.

Objective To explore the relationship of red blood cell distribution width (RDW) with severity and prognosis in children with sepsis. Methods The RDW, clinical features and prognosis of 494 sepsis children were retrospectively ana-lyzed. Results The RDW was increased in 305 sepsis children (61.74%) among whom 49.30%in sepsis group, 60.27%in sever sepsis group and 83.46%in septic shock group. The difference was signiifcant (P<0.001). The mortality of RDW increased children among three groups was 1.89%, 20.45%, 67.57%, and it was signiifcantly different (P<0.001). The rates of increased RDW was 56.15%in 374 survived cases and 79.17%in 120 died cases, and the difference was signiifcant (P<0.001). Pediatric critical illness score (PCIS) was negatively correlated with RDW (P<0.001), while mortality was positively correlated with RDW (P<0.001). Conclusions The rate of increased RDW is high in children with severe sepsis and septic shock and the level of RDW is closely related to the prognosis of patients.

Objective To study ischemic effects on reentrant activities and cardiac arrhythmias using a computational approach. Method The Noble 98 mathematical model of ventricular cell was used in the study. The operator splitting and adaptive time step methods were utilized to integrate the partial differential equations in cardiac conduction models. The ischemic cells were simulated by decreasing the intracellular ATP concentration, reducing the Na+ conductance, and increasing the extracellular K+ concentration in a two-dimensional tissue. Spiral waves were initiated by the cross field technique. Result The results showed that spiral waves in local severe ischemia displayed three different morphologies,whereas in moderate ischemia only two kinds of wave forms exhibited. When the degree of ischemia reached a critical value, the reentrant wave could break. But for larger areas of ischemia spiral wave formed a typical functional reentry around the obstacle. Conclusion The study demonstrates that size and level of ischemia have effects on VTs and VFs. Large ischemic area is beneficial for maintenance of spiral wave and can provide a high probability in the genesis of VTs. Spiral waves can easily break up and degenerate into VFs under critical area or level of ischemia.

Objective:Researching physical-chemic and biologic speciality of VacA-HpaA IgY,which provide a solid experimental basis for the preparation of vaccine against H.pylori infection.Methods:The purified antigen of fusion protein VacA-HpaA was used to immunize hens and the VacA-HpaA IgY was extracted from the yolk by water dilution methods.(1)After the VacA-HpaA IgY was purified by deposition technique with ammonium sulfate,the purity of IgY was analyzed by SDS-PAGE,and protein content of IgY was analyzed by Bradford method.(2)The indirect ELISA was used to detect its heat-stability,acid-stability and pepsin-stability.(3)The specificity of VacA-HpaA IgY to the antigens was identified by Western blot.(4)MTT was applied to assay the neutralization of VacA-HpaA IgY to The cytotoxity of H.pylori.(5)The neutralization of IgY to AGS cells was observed with scanning electron microscopy(SEM).Results:(1)The purity of VacA-HpaA IgY was about 60%,and the protein content of IgY was 22g/L;(2)The IgY showed a good heat-stability,a favorable acid-stability,and a well ability of anti-pepsin digest;(3)Western blot exhibited the protein bands with molecular weight of 27 000 and 30 000.The titer of VacA-HpaA IgY to VacA and HpaA were 1∶3 200 and 1∶6 400;(4)A dose and time dependent correlation of the IgY to counteract the cytotoxic activity of VacA;(5)VacA-HpaA IgY could interrupt the adhesive attraction of H.pylori.Conclusion:The VacA-HpaA IgY has good stabilities and well biologic speciality,which imply that the IgY could be used to prepare treatmental antibody against H.pylori infection.

Objective To explore the mucosal healing quality of hydrotalcite combined with esomeprazole in gastric ulcer.Methods Forty-two patients visiting from June 2014 to December 2015 and diagnosed with gastric ulcer were selected and divided into combination therapy group and single therapy group with 21 patients in each group.The patients of combination therapy group received esomeprazole combined with hydrotalcite,and the patients of single therapy group received esomeprazole alone.The total therapeutic course was eight weeks.At the same period,21 health check-up participants were enrolled as normal control group.The healing of gastric ulcer was observed under white light endoscopy.The morphological changes of gastric pits and microvessel of mucosal at peripheral mucosa around ulcer and normal gastric mucosal were observed under narrow band imaging magnifying endoscopy.The gastric mucosa tissues of the two groups before and after treatment,and normal gastric mucosa of healthy control group were taken.The amount of deposition and composition of collagen fibers,the expression level of factor Ⅷ,the level of transforming growth factor (TGF)-beta1 and the content of hydroxyproline were analyzed by Masson,immunofluorescent and immunohistochemistry staining as well as enzyme linked immunosorbent assay.Chi square test,one-way analysis of variance (ANOVA),least significant difference (LSD) method,Dunnett's T3 and Kruskal-Wallis test and other method were used for comparison.Results After treatment,18 patients of combination therapy group (21 patients) had regular microvessel nets (85.7％),which was significantly more than those of single therapy group (12 cases,57.1％) and healthy control group (10 cases,47.6 ％),and the differences were statistically significant (x2 =4.200,P=0.040).In the comparison of maturity of regenerative mucosa between combination therapy group and single therapy group after treatment,the ration between collagen type Ⅰ and Ⅱ,deposition of collagen fibers,the number of factor Ⅷ positive cells,the level of TGF-beta1 and the content of hydroxyproline were 36.05 and 23.14;269 375.63.± 171 608.63 and 137 693.14±98 330.93;34.91±8.40 and 28.24±6.93;104 498.71±40 487.96 and 70 757.11±19 323.95;(1 897.80±879.35) and (1 230.57±536.05) μg/L,respectively;while in healthy control group,the above parameters were 36.81,245 696.90 ± 224 687.00,23.10 ± 8.40,94 048.04 ±41 306.55 and 1 681.20 ± 423.61 μg/L,and the differences were statitically significant among these three groups (H=7.375,F=3.465,11.680,5.190,5.160;all P＜0.05).Those parameters of combination therapy group were significantly higher than those of single therapy group (H=2.416,LSD method;all P＜0.05).Conclusion Hydrotalcite combined with esomeprazole in the treatment of gastric ulcer could significantly improve microvessel morphology,maturity degree of regenerative mucosal structure and function,and the mucosal healing quality was also superior to single esomeprazole group.