Objective To establish an analytical method to investigate the protective effect of reversed lipid-based nanoparti-cle(RLBN)for topotecan(TPT)in artificial intestinal fluid. Methods Reversed lipid-based nanoparticle of TPT(RLBN-TPT)was prepared by the two-step methods of lyophilization and dissolution. An analytical method was established to determine the concentra-tions of both forms of TPT by high performance liquid chromatography(HPLC). Release curve of RLBN-TPT in simulated intestinal flu-id(SIF)was investigated to study the stability of TPT in gastrointestinal(GI)fluid. Results Both lactone and carboxylate forms of TPT were well separated and determined precisely by the optimized HPLC method. The calibration curves were linear within the range of 0.25-5μg/ml for both forms of TPT. Compared with free TPT,RLBN-TPT significantly improved the stability of TPT in SIF as the per-centage of carboxylate form was remarkably lower than the free TPT(P<0.05). Conclusion RLBN can significantly protect the TPT from hydrolysis in GI,which may lay the foundation for the deuelopment of oral chemotherapeutic drug with higher bioavailability.

Objective: To study the antiemetic effect of the ethyl acetate extract with ethanol extraction of Galangal (hereinafter referred to as galangal extract) in allotriophagic Kaolin model rats.Methods: The rats were administered respectively with chemical drug cisplatin and apomorphine, and received the rotation stimulation as well.Three types of allotriophagic Kaolin models were established, and apomorphine, ondansetron and metoclopramide were used as the control drugs.The experimental rats were divided into 3 groups at low, medium and high dose of Galangal extract with prophylactic effect.The antiemetic effect was observed after the treatment.Results: Chemical drug cisplatin (3.0 mg·kg-1, intraperitoneal injection) and apomorphine (3.2 mg·kg-1,subcutaneous injection) and the rotation stimulation (centrifugal force of 3.4×g,60min) could induce allotriophagic vomiting in rats.The galangal extract at low (1 500 mg·kg-1), medium (4 500 mg·kg-1) and high (9 000 mg·kg-1) dose all fail to effectively inhibit the allotriophagic Kaolin behavior of rats caused by cisplatin, apomorphine and rotation(P>0.05).Conclusion: Galangal extract can not effectively inhibit the vomiting of allotriophagic Kaolin model rats.Probably, there is no correlation between the pharmacological effects of galangal extract and substance P, 5-hydroxytryptamine, dopamine as well as receptive zone of chemical vomiting inhibition.

AAV-ITR single strand DNA mini vector (AAV-ITR ssDNA mini vector) is a novel gene expression vector based on AAV-ITR. We have shown efficient gene expression of AAV-ITR ssDNA mini vector in HEK 293T. Here, we studied the efficacy of gene expression of AAV-ITR ssDNA mini vector in vivo. We injected the skeletal muscle of ICR mice separately with equal molars of AAV-ITR ssDNA mini vector, ITR mutated AAV-ITR single strand DNA mini vector (AAV-ITRmm ssDNA mutant vector), AAV-ITR dsDNA and pUC57-minivector-GFP, combined with TurboFect. Florescence microscope analysis of skeletal muscle section shows that AAV-ITR ssDNA mini vector had higher expression efficiency and longer expression period. We extracted DNA from the muscle three months after injection and quantified three vectors by Real-time PCR. RT-PCR analysis shows that there were highest copy numbers of AAV-ITR ssDNA mini vector existing in muscle. Stable existing of AAV- TR ssDNA mini vector in muscle could be the molecular basis of long term gene expression of the vector. The results suggest that AAV-ITR ssDNA mini vector might be a promising vector for gene therapy.
Animals ; DNA, Single-Stranded ; genetics ; Dependovirus ; genetics ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Mice ; Mice, Inbred ICR ; Muscle, Skeletal ; metabolism ; Real-Time Polymerase Chain Reaction

Objective Human lung adenocarcinoma SPC-A1/DTX cells have a higher radioresistance than SPC -A1cells. This study was to investigate the role of Aurora-An/uclear factor κB ( NF-κB) in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells and its possible molecular mechanisms . Metho ds We collected human lung adenocarcinoma SPC-A1 and SPC A1/DTX cells and divided them into four groups:sh-Aurora-A ( Aurora-A plasmid interference ) , sh-NC, NF-κB inhibition ( SPC-A1/DTX +NF-κB inhibitor ) , and DMSO control .We measured the in vitro radio-sensitivity of the cells by MTT assay , determined their proliferation ability by cloning assay , and detected the mRNA and protein expressions of the target genes by real -time quantitative RT-PCR and Western blot , respectively . Results The 50% effective doses ( ED50 ) of the SPC-A1 and SPC-A1/DTX cells on radiotherapy were (6.5 ±0.3) and (12.8 ±0.6) Gy, respectively, with statisti-cally significant difference between the two groups ( P <0.01 ) .In the radiation doses of 0, 2, 4, and 6 Gy, the numbers of the cloned SPC-A1 cells were 345 ±20 , 252 ±22 , 170 ±15 , and 81 ±10 , sig-nificantly lower than those of the cloned SPC -A1/DTX cells (402 ±21, 370 ±18, 301 ±16, and 252 ±15) (P<0.05).The protein and mRNA expressions of Aurora-A were remarkably higher in the SPC-A1/DTX than in the SPC-A1 cells (1.00 ±0.08 and 1.00 ±0. 06 vs 0.49 ±0.03 and 0.22 ±0.02, P<0.05).MTT assay showed a higher ED50 in the sh-NC than in the sh-Aurora-A cells ([11. 8 ±0.5] vs [7.1 ±0.3] Gy, P<0.01) as well as in the control than in the NF-κB inhibition group ([11.7 ±0.5] vs [6.1 ±0.3] Gy, P<0.01).Inhibition of Aurora-A increased the expression of IκBa by 2.18 ±0.32 times (P<0.01) and that of NF-κB by 0.24 ±0.03 times (P<0.01).The expressions of IκBa (1.00 ±0.05) and NF-κB (1.00 ±0.04) were significantly lower in the parent strains of SPC-A1 than 0.65 ±0.04 and 2.18 ±0.15 in the drug-resistant strains of SPC-A1/DTX (P<0.01). Conclusi on Auro-ra-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells.

Cardiovascular diseases(CVDs) are a major cause of morbidity and mortality in the world.So far,there has been substantial progress toward understanding the pathophysiology and treatment of CVDs.There are multiple cell signaling cascades,some of which are beneficial or compensatory and others deleterious.The balance between these pathways determines the outcome as a diseased or non-diseased state.Protein phosphorylation,which is mediated by enzymes,called protein kinases,is a major mechanism for transducing external stimuli into intracellular signals.Electively targeting of signaling pathways using protein kinase inhibitors would have a potential advantage over receptor blockers.By now,there are types of protein kinase inhibitiors available for treating several diseases.The success of kinase inhibitors in cancer treatment has strongly supported application in the treatment of CVDs.Here,we will review several kinds of protein kinases as potential targets for CVDs and some difficulty in identifying a protein kinase as a putative therapeutic target for CVDs.

Cardiovascular diseases (CVDs) are a major cause of morbidity and mortality in the world. So far, there has been substantial progress toward understanding the pathophysiology and treatment of CVDs. There are multiple cell signaling cascades, some of which are beneficial or compensatory and others deleterious. The balance between these pathways determines the outcome as a diseased or non-diseased state. Protein phosphorylation, which is mediated by enzymes, called protein kinases, is a major mechanism for transducing external stimuli into intracellular signals. Electively targeting of signaling pathways using protein kinase inhibitors would have a potential advantage over receptor blockers. By now, there are types of protein kinase inhibitiors available for treating several diseases. The success of kinase inhibitors in cancer treatment has strongly supported application in the treatment of CVDs. Here, we will review several kinds of protein kinases as potential targets for CVDs and some difficulty in identifying a protein kinase as a putative therapeutic target for CVDs.

Objective To assess the clinical feature of older-onset systemic lupus erythematosus.Methods A retrospective study was carried out.Clinical and laboratory findings were collected from 48 patients with older-onset (＞ or =50 years) SLE as well as 424 patients with younger-onset (＜ 50 years) SLE hospitalized in the First Affiliated Hospital of Zhengzhou University from January 2008 to January 2011.Results Among the 472 hospitalized patients with SLE,the ratio of male to female was 1 ∶ 8.08,and older-onset SLE accounted for 10.17％.No significant difference was observed in the incidence of fever,arthritis,decrease in hemoglobin level,increase in erythrocyte sedimentation rate and reduction in complement C4 level,or the positivity rate of anti-ribonucleoprotein (RNP) or anti-Sm autoantibodies (all P ＞ 0.05) between the older-onset and younger-onset SLE patients.The patients with older-onset SLE showed a statistically higher incidence of myalgia,myasthenia,serositis,heart damage and lung damage,decrease in white blood cell count and platelet count,but a significantly lower incidence of butterfly rash,alopecia,renal damage,Raynaud's phenomenon,photosensitivity,decrease in complement C3 level,as well as the positivity rate of anti-dsDNA and anti-nucleosome antibodies in comparison with those with youngeronset SLE (all P ＜ 0.05).Conclusions It is likely that older-onset SLE has mild and atypical clinical manifestations,with a relatively low positivity rate of specific immunological indices.Hence,clinicians should pay more attention to older-onset SLE so as to avoid the misdiagnosis or missed diagnosis of it.

Objective To study the amount of rhizospheric microorganisms and soil enzyme activity influenced by Paridis Rhizoma in different locations and of different strains. Methods The amount of rhizospheric microorganisms, soil enzyme activity and their correlation were researched through field survey and collection of rhizospheric soil in Paridis Rhizoma cultivated in Three Gorges Reservoir Region and by microbial dilution plate culture method. Results The amount of rhizospheric microorganisms in Paridis Rhizoma from different habitats showed significant differences. The dominant species in soil microflora was bacteria; the second one was actinomycetes; the fewest one was fungus. The variation trend of the amount of rhizospheric microorganisms was not consistent with the variation trend of rhizospheric microorganisms diversity index. The activity of soil phosphatase, invertase and pepsin in Paridis Rhizoma from different habitats varied. The correlation analysis showed that the correlation between the soil enzyme activity and the amount of rhizospheric microorganisms existed. Conclusion Choosing the suitable strains and habitats of Paridis Rhizoma is beneficial to enhancing the amount of rhizospheric microorganisms and soil enzyme activity, which can create good micro-ecological environment for growth and cultivation of Paridis Rhizoma.

This study aims to develop a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of ivabradine and N-demethylivabradine in human plasma, and investigate effects of stable isotope labeled (SIL) internal standard (IS) on ivabradine. The analytes and IS were extracted from plasma by protein precipitation with acetonitrile, and chromatographied on a Capcell PAK C18 (100 mm x 4.6 mm, 5 μm) column using a mobile phase of methanol and 5 mmol x L(-1) ammonium acetate. Multiple reaction monitoring with electrospray ionization (ESI) was used in the positive mode for mass spectrometric detection. The effect of ivabradine isotope peak [M+H+3] + on IS and the effect of SIL IS purity on ivabradine were evaluated. An appropriate concentration of SIL IS was chosen to permit method selectivity and linearity of the assay over the required range. The standard curves were demonstrated to be linear in the range of 0.100 to 60.0 ng x mL(-1) for ivabradine, and 0.050 0 to 20.0 ng x mL(-1) for N-demethylivabradine. The intra and inter day precision and accuracy were within the acceptable limits for all concentrations. Besides, the interaction between IS and ivabradine did not impact the determination of analytes. This method was successfully applied to a pharmacokinetic study of hydrogen sulfate ivabradine sustained release tablets on Chinese healthy volunteers.

Objective:To explore the application effect of the problem-originated clinical medical curriculum (PCMC) and problem-based learning (PBL) on teaching interns in the department of emergency.Methods:A total of 71 interns received in the department of emergency from January 2018 to January 2020 were selected and divided into an observation group ( n=38) and a control group ( n=33) according to different teaching methods. Among them, the control group received traditional teaching, the observation group took PCMC combined with PBL teaching method. The comprehensive ability scores and teaching quality of the two groups of interns were observed, and their satisfaction with teaching and the patient's evaluation on them were recorded. SPSS was used for t test. Results:There was no significant difference in the theoretical test scores, case analysis ability, and operational skills scores between the two groups before the teaching. After teaching, they were all improved, and the improvement effect of the observation group was significantly better than that of the control group [(92.30±4.37) vs. (83.75±3.98); (90.24±5.31) vs. (85.35±4.57); (91.33±5.28) vs. (86.49±4.42), all P<0.05]; the scores of basic job mastery, work attitude, adaptability and doctor-patient communication of the two groups were all improved, and the observation group was higher than the control group ( P<0.05); the satisfaction of the interns and the evaluation results of patients in the observation group were better than those of the control group ( P<0.05). Conclusion:PCMC combined with PBL used in the teaching of interns in the department of emergency can not only stimulate the interns' interest in learning, increase their theoretical knowledge, improve their learning ability and practical ability, but also improve the quality of teaching and improve their clinical resilience.