There were some limitations existed in current teaching mode of diagnostics,including simple evaluation methods,mechanical internship for body sign,tedious internship for laboratory diagnosis and unsustainable interest-stimulating.Interest teaching was proposed in the teaching reform and some concert measures were implemented including interesting inquiry,interesting skill examination,interesting teaching of laboratory diagnostics,simulated case discussion,interesting training of centesis as well as interesting clinical involvement and interesting medical community activities.The interesting teaching contributed to the improvement of students'comprehensive analysis ability,practical ability,adaptability,humanistic quality,English communication ability and doctor-patient communication,which is of grate importance.

BACKGROUND: Decreasing of absorption of zinc from small intestine can induce dermatitis, alopecia, growth and developmental disorder and so on. It is not very clear that how to keep homeostasis when there is low zinc concentration. The discoveries of zinc transporters and relative researches provide new study direction.OBJECTIVE: To study the effect of different zinc concentration on the expressions of divalent metal transporter 1 (DMT1) and human zinc-regulated transporter (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) 4mRNA, and analyze the potential pathway of absorption of zinc from small intestine in low zinc concentration.DESIGN: Blank-controlled experiment.SETTING: Department of Military Hygiene, Navy Faculty, Second Military Medical University of Chinese PLA.MATERIALS: The experiment was accomplished in Department of Military Hygiene, Navy Faculty of Second Military Medical University of Chinese PLA between October 2004 and May 2005. Materials were human colon adenocarcinoma cells Caco2 that were purchased from Shanghai Institute of Cell of Chinese Academy of Sciences.Time-dependent effect: The expression of DMT1 and ZIP4 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) at 0,dependent effect: The expression of DMT1 and ZIP4 mRNA were measured after the 0, 2.5, 5, 7.5 and 10 μmol/L TPEN exposure, respectively, by RT-PCR.MAIN OUTCOME MEASURES: Time-and dose-dependent effect of zinc on the expression of DMT1 and ZIP4 mRNA in Caco2 cells.RESULTS: ①Time-dependent effect: Compared with 0 hour, the expression of DMT1 mRNA obviously increased at 6, 8 and 10 hours (P ＜ 0.05 ), but there was no significant change at 2 and 4 hours. The expression of ZIP4 mRNA markedly increased in all timing, and ZIP4 mRNA level at the 6th hour reached the peak with the prolongation of low zinc duration,which was 2.1 times ofthat at 0 hour. ②Dose-dependent effect: The expression of DMT1 mRNA distinctly increased at the concentration of 7.5 and 10 μmol/L TPEN exposure as compared with that of the control group (P ＜ 0.05), but there was no significant change at 2.5 and 5 μmol/L. The ZIP4 mRNA expression increased with the increasing of the concentration of TPEN, the expression of ZIP4 mRNA was 2.7 times of that at 0 μmol/L. CONCLUSION: Zinc can regulate the expression of DMT1 and ZIP4 mRNA in Caco2 cells, and there is time-and dose-dependent effect. But the mRNA expression of ZIP4 is more sensitive and prompt than DMT1. Cells can upregulate the expression of DMT1 and ZIP4 mRNA to keep the homeostasis in low zinc condition.

Objective:To study the concentration response and time course of zinc transporter 1 (ZnT 1) and metallothionein (MT1/MT2) mRNA,as well as the cell viability to zinc exposure in primary cultured new born rats hippocampal neuron s. Methods: The cell viability were determined by trypan blue s taining at various concentrations of Zn 2+ (0, 50, 100, 125, 150, 175, 200 ?mol/L). The expression of ZnT 1, MT1 and MT2 mRNA to various concentrations of zinc exposure(0, 50, 75, 100, 125, 150 ?mol/L) and to 100 ?mol/L zinc exposure for different times (0, 2, 4, 6, 8 h) were determined b y real time fluorescent quantitative PCR. Results: The viabilit y of the neurons decreased significantly after 48 h once the concentration of Zn 2+ exceeded 125 ?mol/L,and the expression of ZnT 1 mRNA was in proportion to the increment of zinc.The expression of MT1/MT2 mRNA reached a plateau when the zinc concentration exceeding 75 ?mol/L. The expression of ZnT 1 mRNA peaked on about 2 h.However, the expression of MT1/MT2 mRNA reached its maximal around 6 h at the concentration of 100 ?mol/L. Conclusion: These results imply that although both MTs and ZnT 1 can attenuate the zinc toxicity, they may play different roles at different phases. ZnT 1 enhance the efflux of zinc prior to the sequesteration by MTs. Th e action of ZnT 1 may be durable, but the role of MTs may be satiable.

In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g^-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.

Objective To search a new,safe and effective method to obtain semen using rectal Drobe electroejaculation(RPE).Methods RPE procedures were performed in 8 men with psychological ejaculatory failure,used the electrical stimulate instrument(CGS model,made in Italy).The average age was 26 years.Seminal vesicle was normal by B Uhrasonography or CT examination.Except for 1 case of diabetes,the other 7 cases found no hormones,blood,biochemical and nervous system abnormalities.Three cases had nocturnal emission,but no sexual intercourse ejaculation.Results Adequate sperm was successfully taken out from the 8 patients.Mean seminal fluid volume was 2.3 ml,mean total sperm count was 67×106/ml and mean total motile sperm was 21.3%.Three patients felt abdominal distension during the RPE course.Conclusion RPE is a safe,reliable,non-in-vasive,repeatable method to obtain semen.

Objective The role of long non-coding RNA Linc00467 in human lung adenocarcinoma is not yet clear.This study was to investigate the expression of long non-coding RNA Linc00467 in human lung adenocarcinoma, its clinical significance, and the effects of Linc00467 on the functions of the tumor and endothelial cells in vitro.Methods Lung adenocarcinoma tissue and normal tissue surrounding the malignance were obtained from 60 patients with pathologically proved stage I-Ⅲa lung adenocarcinoma.Human umbilical vein endothelial cells (HUVECs) were transfected with the over-expressed plasmid pccl-Linc00467 (HUVEC experimental group) or the empty vector pccl (HUVEC control group), A549 cells with Linc00467-siRNA (A549 experimental group) or negative siRNA (A549 control group), and H1299 cells, too, with Linc00467-siRNA (H1299 experimental group) or negative siRNA (H1299 control group).The expression level of Linc00467 in the lung adenocarcinoma tissue was detected by qRT-PCR with an analysis of its correlation with the clinicopathological characteristics of the patients;the influence of Linc00467 on the proliferation of the A549, H1299 and HUVEC cells was assayed with CCK-8;and the role of Linc00467 in the angiogenesis of the HUVECs was assessed by fibrin bead sprouting assay.Results The expression of Linc00467 in the lung adenocarcinoma tissue was 2.72±1.31 times as high as that in the normal lung tissue (P<0.01), and those in the A549 and H1299 cells were 3.45±0.25 and 3.22±0.33 times as high as those in the human bronchial epithelial (HBE) cells (P<0.01).The expression level of Linc00467 was significantly correlated with the tumor size and vascular invasion (P<0.05).After transfection of Linc00467-siRNA, the expressions of Linc00467 in the A549 and H1299 experimental groups were down-regulated by 72% and 68% as compared with those in the A549 and H1299 control groups (P<0.01).The number of living cells was remarkably decreased in the A549 experimental group in comparison with the A549 control at 48 h (1.29±0.07 vs 1.51±0.09), 72 h (1.53±0.15 vs 2.13±0.11), and 96 h after culturing (1.98±0.18 vs 3.02±0.12), and so was it in the H1299 experimental versus the H1299 control group, but markedly increased in the HUVEC experimental versus the HUVEC control group (P<0.05).At 5 days, HUVEC experimental group, as compared with the HUVEC control, showed a significantly increased number of newly formed vascular branches (7.36 vs 4.25/superbead, P<0.01) and relative length of the blood vessels (3.12 vs 1, P<0.01).Conclusion Linc00467 promotes tumor cell proliferation and angiogenesis and is highly expressed in the lung adenocarcinoma tissue, which is correlated with the tumor size and vascular invasion and suggests that Linc00467 could be a potential biomarker and therapeutic target.

AIM: To determine whether the high mobility group protein I (HMGI) is able to bind to the upstream sequence of platelet-derived growth factor B-chain gene and to characterize the HMGI-binding AT-rich regions. METHODS: Recombinant human HMGI (rhHMGI) protein was prepared and electrophoresis mobility shift assay (EMSA) was used. RESULTS: The binding of rhHMGI to PDGF-B (-1 758 / +43 bp) was observed in vitro. Two major HMGI-binding fragments -1 392 / -1 180 bp and -188 / +43 bp were identified, which contained the same AT-rich sequence TTTATAAA (-1 333 / -1 326 bp, -1 314 / -1 307 bp and -30 / -23 bp). An oligonucleotide bound to the TTTATAAA and the GAGACC, the core sequence of the shear stress response element of the PDGF-B, could also bind to the HMGI. Furthermore, HMGI facilitated the binding of NF-?B to the GAGACC in the oligonucleotide. CONCLUSION: The HMGI could bind to the upstream sequence of the PDGF-B gene via the AT-rich sequence TTTATAAA, which may play a role in the transcriptional regulation of the PDGF-B gene.

Objective:To investigate the possibility of using radioiodine labeled framework region(FR)antisense oligonucleotides(ASONs)as an imaging agent or antisense therapeutic radiopharmaceu-tical in lymphoma.Methods:A 18-mer partial phosphorothioate oligonucleotide sequence was synthe-sized and grafted in 5'with a tyramine group which was further radioiodinated.Radioiodination of the tyra-mine derivatized oligonucleotides was performed using the chloramine T method.(1)Normal CD-1 micewere injected via a tail vein with 148 kBq (125)~I-FR-ASON(2-3?g).Animals were sacrificed at the endof 1,2,4 and 24h,and tissue samples were studied.(2)Liposome-mediated 3.33 MBq (131)I-FR-ASON(7-9?g)were injected intralumorally into tumor-bearing BALB/c mice(6 weeks after innculation of10~7 Namalwa cells)meanwhile liposome-mediated (131)~I labeled sense oligonucleotides served as controls.Biodistribution was monitored by sequential scintigraphy and organ radioactivity measurement 24h afterinjection.Percentage of the injected dose per gram of tumor and tumor/non-tumor tissue ratios(T/NT)were calculated tot each group of mice and the difference between two groups was assessed.Results:The5′tyramine group allowed specific and stable radinlabeling of the ASON with radioiodine.The radioactivi-ty reached its peak 1h after injection,and then decreased rapidly in normal mice after intravenous ad-ministration of (125)~I-FR-ASON.The liver,stomach and intestine played an important role in biodistributionand radioactivity counts were low in bone,brain and blood.When (131)I-FR-ASON was injected intratumor-ally into mice grafted with Namalwa cell line,images showed the tracer accumulated in the tumor,Imme-diately after intratumoral administration,only the tumor was visible.Scintiscans performed at the end of 1and 2h showed elimination of the tracer from the tumor to the abdomen and at the end of 24h the tumorwas clearly seen.Percentage of the injected dose per gram of tumor and T/NT ratios for the sense group(control)were significantly lower than those of the antisense group.Conclusion:Radiolabeled Ig FRASON showed high specificity in V1 family B-cell lymphoma,which should be further investigated for nu-clear medicine imaging application and radionuclide antisense therapy.

The human venous valve of the brachial,femoral and long saphenous veins wereexamined with light,transmission and scanning electron microscopy.The observationshows that the venous valve is composed of three functional layers covered withendothelium on both surfaces.A loosely structured layer is located underlying theendothelium.A network mainly containing randomly oriented elastic fibers was foundnear the side towards the lumen.To the side towards the venous wall,there is adense layer composed of eircumferentially and transversly oriented collagen bundles.Some smooth muscle cells extend from the wall of the vein to the base of thevenous valve.The elastic fibers and smooth muscle cells together with the collagenfibers contribute to the mechanical load-bearing performance of the valve and to thepassive closing and openning mechanism.In addition,the smooth muscle cells mightplay an active role in the normal functioning of the valve.The scanning and transmission electron microscopy of venous valve showdifferent arrangement of the endothelium.On the surface of the valve next to the wallof the vein,the endothelial cells are transversely arranged,while on the othersurface over which the current of blood flows,the cells are longitudinally arrangedin the direction of the current.These accord with the role of fluid mechanics.12 normal venous valves were tested by universal testing instrument (Instrontype 1122).The mean value of the maximum tension of the valve is 1 N.Theaverage value of the tensile ultimate strength is 10N/mm~2.

This paper introduces the structure, contents, function of the digital teaching re-courses database in Clinical Skills course of Central South University, analyses the essence and char-acter of the flipped classroom (FC), and emphasizes flexible environment, learning culture, inten-tional content, professional educator in its implementation process. Meanwhile, it introduces the appli-cation of flipped classroom based on digital resources database in the teaching of Clinical Skills course, mentions the importance of the four aspects:preparing teaching materials, learning before class, guidance and cooperation in class, evaluation and knowledge expansion after class. Lastly, it analyses the advantages and foreground of it.