A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

China ; epidemiology ; Communicable Diseases ; epidemiology ; Disease Notification ; methods ; Humans ; Mandatory Reporting ; Rural Health Services

This paper points out the experimental animal ethics education in pharmacology teaching and the im-portance of the experiment,it is good to raises the student good humanity accomplishment and scientific research quality, and promote medical research and life ethics of benign interaction.It also expounds the basic content of ex-perimental animal ethics education,including:animal welfare, the 3R principle, AAALAC accreditation and analy-sis of the experimental animal welfare legislation status, raises questions about animal ethics education problems and thinking in the pharmacology experiment teaching in ourschool.The experimental animal ethics education should become an important part of pharmacology experimental teaching, which enhanced the students′s ethical awareness to better understand and respect for life, and contribute to the sustainable development of medical and pharmaceutical research.

BACKGROUND: Conventional hemodialysis mainly for cleaning uremic micro molecule substance, such as urea nitrogen or creatinine; however, few hemodialyses can clean uremic middle molecule substances (MMS). With prolonged dialysis duration, MMS accumulates in vivo and induces a series of complications. OBJECTIVE: To compare the efficiency of adsorptive dialysis (hemoperfusion unites hemodialysis) and conventional hemodialysis in cleaning uremic MMS. METHODS: Totally 60 maintenance hemodialysis patients were averagely divided into the adsorptive dialysis group and conventional hemodialysis group. First of all, hemoperfusion apparatus and dialyser were connected in series to take the adsorptive dialysis in the adsorptive dialysis group (hemoperfusion apparatus were equipped before dialyser). 120 minutes later, the hemoperfusion apparatus was toke off and continues to hemodialysis for 120 minutes. Duration of conventional hemodialysis was 240 minutes. Changes in clinical symptoms and levels of liver function, kidney function, serum electrolytes, hemocytes and uremic MMS were observed prior to and after treatment. RESULTS AND CONCLUSION: Adsorptive dialysis could remove the MMS notably. Compared with the conventional hemodialysis group, a single 120 minutes treatment could decrease MMS significantly (P < 0.05). The platelet levels were obviously decreased in the adsorptive dialysis group after treatment (P < 0.05), which were significantly different from the conventional hemodialysis group (P < 0.05). There was no significant difference in liver function, kidney function or serum electrolytes concentration. But related symptoms, such as the skin itch, sleep disorders and myalgia, were relieved more or less.

A gene that could be potentially involved in spermatogenesis was identified and characterized by using suppression subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE) with total RNA from type A spermatogonia and pachytene spermatocytes of rat. This gene consists of 3 433 base pairs (bp) with a complete open reading frame (ORF) of 3 171 bp and encodes a putative protein containing 1057 amino acids. The nucleotide sequence displays a 93% identity to mouse ubiquitin-activating enzyme E1, Chr Y 1 (Ube1y1) and an 82% identity to human ubiquitin-activating enzyme E1 (UBE1). The putative protein of this gene contains an ubiquitin-activating enzyme signature 1 and an ubiquitin-activating enzyme active site, which are also existed in mouse ubiquitin-activating enzyme E1, human ubiquitin-activating enzyme E1 et al. So we named this gene as Rattus norvegicus ubiquitin-activating enzyme E1 (Ube1). The sequence of Ube1 was submitted to GenBank and the accession number is EF690356. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that Ube1 was specifically expressed in testis, while its expression was not detected in heart, brain, spleen, lung, liver, muscle, kidney and ovary. Comparison of the expression of Ube1 in different developmental stages of testis and Sertoli cells (real-time PCR) indicated that Ube1 was expressed more highly in spermatogonia than in spermatocytes, spermatids and Sertoli cells. In conclusion, Ube1 is a gene encoding rat ubiquitin-activating enzyme E1 and specifically expressed in testis, which might play a key role in ubiquitin system and influence spermatogenesis.
Animals ; DNA, Complementary ; genetics ; Male ; Rats ; Real-Time Polymerase Chain Reaction ; Spermatids ; metabolism ; Spermatocytes ; metabolism ; Spermatogenesis ; genetics ; Spermatogonia ; metabolism ; Testis ; metabolism ; Ubiquitin-Activating Enzymes ; genetics ; metabolism

Objective To investigate the children's body environmental Se and T-2 toxin level in their staple food in Kaschin-Beck disease(KBD)relative active regions in Aba state of Sichuan province in 2008.Methods We took X-ray photograph of the right hand on children aged 7-13 years in 48 villages from 11 counties in Aba state.The relative active regions of KBD were chosen according to the X-ray result and historical status of KBD.The children's urine and hair,drinking water and their staple food werr sampled.Selenium contents in urine,hair,water and food samples were determined by naphthalene fluorescence,and T-2 toxin in staple food samples were detected by ELISA kits.Results In 2145 X-ray films,66 films were positive,and the children's KBD positive rate was 3.08%(66/2145).The KBD positive rate was respectively 10.98%(29/264)and 8.52%(19/223)in Maerkang county,Jinchuan county and it was 0.75%(3/400)in Rangtang county,historically serious endemic area.The selenium content in urine of children aged 7-13 years in Maerkang county,Jinchuan county and Rangtang county was (10.41±4.67), (10.11±3.65), (8.42±2.68)μg/g Cr, respectively, there was no statistical difference among three counties(F=0.901, P>0.05). The selenium content in hair of children aged 7-13 years in Maerkang county[(0.18±0.04)mg/kg] was lower than that in Jinchuan county[(0.21±0.04)mg/kg, P<0.05].The selenium content in water in Jinchuan county [(0.225±0.124 )μg/L ] was lower than that in Maerkang county and Rangtang county[(0.320±0.092), (0.339±0.105)μg/L, all P<0.05]. The selenium content in staple food in Jinchuan county(0.0033 mg/kg) was lower than that in Maerkang county and Rangtang county(0.0258,0.0137mg/kg, Z=-6.146,-3.042, all P<0.017). The T-2 toxin level in flour in three counties was 19.60,17.95,26.25 ng/g,respectively,there was no statistical difference among three counties(X2=5.623, P>0.05).The T-2 toxin level in grain Maerkang county (10.72 ng/g) was higher than that in Jinchuan county and Rangtang county (3.74,3.30 ng/g, Z=-6.315,-4.407,all P<0.017). T-2 toxin contamination in flour was more severe than that in grain (Z=-6.690,-5.493,-3.676, all P<0.05). Conclusions In 3 relative active KBD regions of Aba state,the children's selenium nutritional status and the T-2 toxin contamination level in their staple food is consistent with the distribution of KBD.

The emerging nano-black phosphorus materials have created a new platform for biomedical research. Nano-black phosphorus has the following advantages: black phosphorus can produce singlet oxygen under near-infrared light irradiation, so it can be used as a photosensitizer for photodynamic therapy;black phosphorus has extensive light absorption in the long wavelength region, and this near-infrared photothermal property can be used in photothermal therapy. The high specific surface area and unique fold structure of the black phosphorus nanosheet make it have very high drug loading.This paper mainly reviews the applications of black phosphorus in biological imaging, photothermal therapy, photodynamic therapy, and as a drug carrier in recent years. Based on the photoelectric properties of black phosphorus nanomaterials combined with intelligent drug delivery platform, the synergistic effects of light/heat/chemistry, light/chemistry/gene, and light/chemistry/immunity can be produced, which has a broad application prospect.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.
Administration, Intravaginal ; Adult ; Antifungal Agents ; blood ; chemistry ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; methods ; Female ; Humans ; Imidazoles ; administration & dosage ; blood ; pharmacokinetics ; Middle Aged ; Molecular Structure ; Reproducibility of Results ; Tandem Mass Spectrometry ; methods ; Time Factors ; Young Adult

Aim To explore the therapeutic effects of main active compounds of panaxadiol ( PD ) in on Alzheimer’s disease ( AD) via network pharmacologi-cal analysis and Mmolecular docking. Methods A to-tal of 107 prescriptions for AD treatment were screened by using network pharmacology, screening for the high-est frequency of ginseng and its target for AD. Use mo-lecular docking technology was used to find components with the highest score for non-receptor tyrosine kinase ( FYN) docking. Then we successfully estimatedestab-lished AD cell model with overexpressinged APP pro-teins in vitro. Next,the cell viability was detected by MTT assay,the cell damage was detected by LDH as-say,the apoptosis and intracellular Ca2+concentration were detected by flow cytometry, and phosphorylated FYN protein expression was detected by Western blot detection of . phosphorylated FYN protein expression. Results Eighteen active components of Gginseng and 29 AD-related targets were screened by the method of network pharmacology. The results of molecular doc-king showed that PD had strong binding effects with FYN. The results showed that PD could increase the survival rate of cells,reduce the release of LDH,reduce apoptosis,and improve AD cells’ intracellular Ca2+o-verload and reduce the expression of FYN-Y416 pro-tein. Conclusion The experimental results of network pharmacology were are verified and the protective effect of PD on AD may be related to inhibition of FYN signa-ling pathway.

The purpose of this research is to study the effect of small molecule compound piceatannol (PIC) on host inflammation in adenine induced chronic kidney disease (CKD) mice, and then to explore its mechanism based on the regulation of gut microbiota. All procedures were approved by the Institutional Animal Care and Use Committee of the Nanjing University of Chinese Medicine. The level of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected by enzyme linked immunosorbent assay (ELISA); UPLC-TQ/MS technology was used to monitor the level of proinflammatory uremic toxin indoxyl sulfate (IS) and p-cresol sulfate (PCS); the expression of occludin was tested by Western blot; in vitro anaerobic culture of gut bacteria was used to produce indole; the abundance of gut microbiota was evaluated by 16S rDNA sequencing. The results showed that PIC had no effect on inflammatory infiltration in kidney tissue of CKD mice, but could decrease IL-6 level in blood and IL-6/TNF-α level in colon tissue. PIC did not improve intestinal occludin protein expression in CKD mice; while it could significantly reduce the levels of IS and PCS in blood and liver of CKD mice. Further mechanism studies showed that PIC could inhibit the synthesis of IS precursor indole in gut bacteria. Moreover, PIC could decrease the abundance of gut bacteria which producing uremic toxin, such as reducing the abundance of indole and p-cresol producing gut bacteria. In conclusion, PIC could regulate gut microbiota and inhibit the synthesis of uremic toxin precursor, thereafter reducing the accumulation of IS and PCS in vivo, ultimately relieving the inflammation of CKD mice.