From teaching design principle, the article has discussed the key element of the process of hygiene examination teaching designs, and has brought forward three kinds of rational science design process patterns.

With the extension of university development and large-scale expansion,the connotation building is related to the sustainable development and stamina of colleges and universities,so colleges and universities begin to shift the focus on the "scale effect"gradually to "content-building". This paper analyzes the important role the connotation building plays in the development of universities from the need of university history,disciplines,professional develop-ment and personnel training.

Combined with the situation of preventive medicine teaching in adults this article discusses the teaching mode of preventive medicine in adult-upgraded education from talent training plan revision,teaching contents and teaching system reform,and practicing teaching mode construction,to meet the demand of development of modern public health.

The fragmentation pathways of two taxanes drugs have been studied in positive ion mode by Q-TOF with the advantages of high mass accuracy and high resolution analysis. The [M+H] + ions were observed by ESI-MS, from which the molecular weights were obtained. Using the protonated pseudo-molecular ions [M+H]+ as internal reference compounds, the accurate mass and element composition of the fragment ions were determined. The collision induced dissociation (CID) data of the [M+H] ions provided fragmentation pathways of related compounds. Results showed that the major cleavage pathways of paclitaxel and docetaxel were the same that the cleavage of C-O bond between the side chain and taxol skeleton easily occurred, then stripping of the functional groups on the parent ring. Some common fragments were formed, such as m/z 105.033 7, 291.137 3, 309.148 5, 327.159 7, 387.181 2 and 509.217 4, which would provide a basis for future qualitative and quantitative analysis of taxanes in vitro and in vivo.
Paclitaxel ; chemistry ; Peptide Fragments ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Taxoids ; chemistry

With the new talents demand model set,quality education has inevitably become the central content of higher education.Based on an experience of 21 years in talents majoring in preventive medicine cultivation,we put forward exploratory thoughts to improve the comprehensive quality and social adaptability of college students,which included the construction of teaching staff, the practice of tutorial system for medical undergraduate students,the innovation in teaching practice and the establishment of stimulating mechanism in talent cultivation.

Objective To evaluate the expression of human vascular endothelial cell growth factors 165 (hVEGF165) gene transfected into fibroblasts by recombinant adenovirus and study the repairing effect of this cells on radiated skin ulcer in rats.Methods The recombinant adenovirus with hVEGF165 was established and transfected to rat primary fibroblasts, and its expression of hVEGF165 in fibroblasts was identified with real-time PCR, immunocytochemistry and Western blot.Twenty four clean grade SD rats of were irradiated locally with 50 Gy γ rays to generate an animal model of radiation skin injury.The hVEGF165-transfected cells were injected to the irradiated site under rat skin 7 d post-irradiation.The therapeutic effects on the irradiated skin wound were evaluated through general observation as well as histological staining of HE.The expression of hVEGF in the irradiated skin tissue with fibroblasts injection was analyzed by Real-time PCR.Results The hVEGF165 gene was overexpressed in the transfected cells and approached to 88 373-fold bigger compared to controls transfected with blank vectors, and an extensive expression of VEGF in the cytoplasm of transfected cells was observed by immunohistochemistry.VEGF protein with the relative molecular mass of 23 000 was also detected in cell lysate by Western blot.The local skin ulcers in rats occurred about two weeks after irradiation.In the hVEGF165-transfected group, the average area of radiation-injured skin was 40.2 mm2, about 57％ less than that of the control group transfected with blank vectors so that the healing time was shorten by 6 days.The relative concentration of hVEGF mRNA in the skin tissue of rats injected with hVEGF165-transfected cells were 5.15-fold and 4.15-fold bigger compared to that of controls (t =3.385,3.220, P ＜ 0.05) at 3 and 7 d after administration.Conclusions The primary fibroblasts transfected with hVEGF165 gene could efficiently release VEGF to the irradiated skin tissue and promote the recovery of irradiation skin ulcers by shortening the healing time and thus enhanced the therapeutic effect on skin wounds.

Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) in subarachnoid space on chronic neuropathic pain in rats. Methods One hundred and forty female SD rats weighing 180-200 g were used in this study. Chronic neuropathic pain (NP) was induced by ligation and separation of tibial and common fibular nerves (SNI). Two weeks after the surgery the animals were randomly divided into4 groups (n=35 each):group Ⅰ NP; group Ⅱ NP+ MSC (MSCs); group Ⅲ NP+ phosphate buffer saline (PBS) and group Ⅳ NP+ bone marrow monocyte (BNMCs). MSCs 10 μl, PBS 10 μl and BNMCs 10 μl were injected into subaraclmoid space at 2 weeks after surgery in group Ⅱ , Ⅲ and Ⅳ respectively. Paw withdrawal threshold to yon Frey filament stimulation (PWT) was measured before surgery (T0, baseline), at 2 weeks after surgery (T1) and 1, 2, 3, 4 and 5 weeks after subarachnoid injection (T2-6). Five animals were killed at T1-6 in each group and their lumbar enlargements were removed for determination of BDNF mRNA expression. Results PWT was significantly decreased by SNI at T1 in all 4 groups, and at T2-6 in group Ⅰ , Ⅲ and Ⅳ as compared with the baseline at T0 (P ＜ 0.05). Subarachnoid MSC transplantation significantly reduced mechanical hyperalgesia at T2-6 and up-regulated BDNF mRNA expression at T2-4 as compared with that at T1 (P ＜0.05). There was no significant change in PWT and BDNF mRNA expression after subarachnoid PBS and BNMCs injection in group Ⅲ and Ⅳ (P ＞ 0.05). Compared with group Ⅰ , PWT was significantly increased and BDNF mRNA expression was up-regulated in group Ⅱ (P ＜ 0.05), but no significant change in PBS and BNMCs was found in group Ⅲ (P ＞ 0.05) .Conclusion Up-regulation of BDNF mRNA expression in the spinal cord may be involved in the amelioration of chronic neuropathic pain by subarachnoid bone marrow mesenchymal stem cell transplantation.

With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.
Enzymes ; chemistry ; Humans ; Inactivation, Metabolic ; Mass Spectrometry ; Membrane Transport Proteins ; chemistry ; Pharmacokinetics ; Proteomics

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

As an emerging discipline,the development of medicine verification with sanitary verification direction promotes the convergence of China's public health cause and the international’ s powerfully. Associated with our experience in teaching practice,we grope after the best way to foster students’innovative capability and promote this discipline’s development in the aspects of course content,curriculum system,experimental tuition,practical capability etc.