We have carried out experiment design and comment,and students write the reports of experiment design in patho- physilolgy teaching from the aspects of basic process of experiment research and basic factors,principle and meaning of experi- ment design.By way of the teaching reform,the major position of students in studying is established,students' ability to study in- dependently and acquire knowledge actively are well cultivated,their comprehensive quality are enhanced and the teachers con- struction is also promoted.

To study the toxicokinetics of bakuchiol, hepatic and renal toxicity in rats after single oral administration of Psoraleae Fructus and combined with Glycyrrhizae Radix et Rhizoma, in order to provide scientific evidences for clinical safe medication use. A total of 35 SD rats were randomly divided into seven groups: vehicle (distilled water) control group, Glycyrrhizae Radix et Rhizoma group, positive control (aristolochic acid A) group, Psoraleae Fructus (40 g x kg(-1)) group( both male and female rats), Psoraleae Fructus and Glycyrrhizae Radix et Rhizoma (40 +20) g x kg(-1) group (both male and female rats). HPLC-UV method was used to determine the concentration of bakuchiol in rat plasma at different time points after single oral administration. Plasma alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), plasma creatinine (Cr), N-acetyl-β-D-glucosaminidase (NAG) and kidney injury molecule 1 (Kim-1) were measured after administration for 24 h. The main toxicokinetics parameters of bakuchiol in rats exert significantly gender difference. When Psoraleae Fructus combination with Glycyrrhizae Radix et Rhizoma, the total area under the plasma concentration-time curve( AUC), C(max), and plasma clearance (CL) of bakuchiol were increased, respectively; CL, half-life (t½) were decreased, and T(max) were prolonged. The biochemical indicators (including ALT, AST, BUN, Cr and KIM-1 level) in different dose of Psoraleae Fructus groups, were found no statistically significant difference when compared with vehicle control group. The level of NAG in both Psoraleae Fructus and compatibility with Glycyrrhizae Radix et Rhizoma groups were significant increased (P < 0.05). There are obvious effects on toxicokinetics of bakuchiol in rats when Psoraleae Fructus combined with Glycyrrhizae Radix et Rhizoma. Renal toxicity induced by Psoraleae Fructus at high dose was observed after single oral administration and no liver damage in rats was found.
Administration, Oral ; Animals ; Female ; Glycyrrhiza ; toxicity ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Phenols ; pharmacokinetics ; toxicity ; Psoralea ; toxicity ; Rats ; Rats, Sprague-Dawley ; Rhizome ; toxicity ; Toxicokinetics

Objective:To analyze the activities of human NKG2D ligand MICA/B promoter induced by 5-Fu.Methods:The 5'-end flanking regions of MICA /B promoter and their different truncated fragments were amplified from A549 genome by PCR. The resulting amplicons were cloned into pGL3-Basic vector to generate the MICA/B luciferase reporter plasmids. All the constructs were transiently transfected A549 cells. The promoter region activities were determined by dual-luciferase reporter assays. The effect of 5-Fu on the promoter activities of MICA/B was also tested.Results and Conclusion:The 5'-end flanking regions of MICA /B promoter and five of their different truncated fragments were successfully obtained. The normalized luciferase reporter gene activities driven by the above promoters and fragments were 3.61,2.26,1.63,0.313,0.711 and 0.663 for MICA and 17.49,10.11,7.398,0.822,0.997 and 0.49 for MICB,respectively. Promoter activities in transiently transfected A549 cells treated by 20,40,80,160 and 320 μg/m of 5-Fu increased 1.69,1.48,1.62,1.55 and 1.78 fold for MICA and 1.44,1.87,1.38,1.19 and 1.25 fold for MICB. Our results suggest that 5-FU can significantly up-regulate the promoter activity of both MICA and MICB.

Objective To study the personality characteristics of the stagnation of liver-qi syndrome cases with Minnesota Multiphasic Personality Inventory (MMPI).Methods 20 liver-qi stagnation syndrome cases and 20 normal persons whose paired by sex and age were tested by MMPI. The scores of 13 clinical scales and the section plane of MMPI were acquired and analyzed. Results There was a significant difference in 9 clinical scales (F, Pa, D, Pt, Si, Sc, Hs, Hy, Pd) between the liver-qi stagnation syndrome cases and normal persons ( P<0.05～0.001). The scores of Hs, D, Hy, Pd in the liver-qi stagnation syndrome group were higher than the Chinese normal model. The MMPI section plane of liver-qi stagnation group presented the type of 3/1. Conclusion The patients with liver-qi stagnation have some special personality characteristics such as depression, anxiety, loneliness, indifferent and attention.

Objective To study on the effects of Huaweishu granule on cisplatin-induced Beagle dog vomiting models and its mechanism.Methods Cisplatin-induced Beagle dog vomiting models were adopted as research object.The changes of vomiting frequency,latency,serum motilin content in the medulla oblongata and ileum,5-HT and substance P content of these models after using Hua weishu Granule were observed.Results ① Latent period of vomiting of the model group was (60.8±37.1)min; while this period was (137.3± 53.4)min,(122.8 ± 50.7) min,(l16.8±44.6)min,in the Huawei-shu low,medium and high dose group respectively,all showing a statistic difference than the model group (P＜0.05).②Frequency of vomiting in the model group was (270.3±51.8),while this frequence was (111.5±45.0) and (149.5±26.8) in Huawei-shu low and medium dose group respectively; ③ serum motilin in the model group was (0.354±0.098)ng/ml,while serum motilin in Huawei-shu low,medium and high dose group was (0.230±0.074) ng/ml,(0.235± 0.071) ng/ml,and (0.245± 0.062)ng/ml respectively,all lower than the model group (P＜ 0.05).④ Medulla oblongata 5-HT in the model group was (2.028 ±0.198)ng/ml,while medulla oblongata 5-HT in Huawei-shu low,medium and high dose group was (1.620±0.329)ng/ml,(1.194±0.386)ng/ml,and (1.269 ± 0.251) ng/ml respectively,all were lower than the model group (P＜0.05); ileum 5-HT in the model group was (1.634± 0.221)ng/ml,while ileum 5-HT in Huawei-shu low,medium and high dose group was (1.108±0.291)ng/ml,(1.194±0.386)ng/ml,and (1.269 ± 0.251) ng/ml respectively,all were lower than the model group(P＜0.05) ;⑤ ileal substance P content of the model group was (0.356±0.063)ng/ml,while ildeal P content in Huawei-shu low dose group was (0.274±0.064)ng/ml,lower than the model group than(P＜0.05).The medullary substance P content was (0.432±0.021)ng/ml in the model group,while medullary P content in Huawei-shu low,medium and high dose group was (0.370±0.040) ng/ml,(0.385±0.029) ng/ml,and (0.386± 0.041)ng/ml respectively,all were lower than the model group(P＜0.05).Conclusion Huawei-shu granule can prevent cisplatin-induced Beagle vomiting.

OBJECTIVETo observe the effect of Astragalus-Angelica Mixture (AAM) on osteopontin (OPN) expression in rats with chronic nephrosclerosis.

METHODSChronic nephrosclerosis model rats induced by repeated intraperitoneal injection of puromycin were randomly divided into the model group, AAM group and Irbesartan (an antagonist of angiotensin) group. The experimental course lasted 12 weeks. Blood and urine samples were examined by biochemical method. Kidney tissue was taken for pathological stain and immunohistochemical method and was applied to examine OPN expression, mononuclear macrophage, laminin in extracellular matrix and decorin expressions.

RESULTSAAM showed the effects of decreasing urinary protein and improving renal function similar to that of Irbesartan. It also could alleviate the pathological damage of kidney tissue, especially in decreasing renal tubular mesenchymal damage index. The accumulation of decorin and laminin in the mesenchymal extracellular matrix significantly decreased. Renal tubular OPN expression and mesenchymal infiltration of mononuclear macrophage decreased significantly and in a positive correlated manner (r = 0.885, P < 0.01).

CONCLUSIONAAM has similar renal protective action to that of Irbesartan, this action may be related to the inhibition of up-regulated OPN expression.

Animals ; Astragalus Plant ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Nephrosclerosis ; chemically induced ; metabolism ; Osteopontin ; Phytotherapy ; Puromycin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; biosynthesis

Animals ; Blood Pressure ; Lymphatic Vessels ; physiopathology ; Male ; Norepinephrine ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; physiopathology

OBJECTIVETo construct two recombinant lentiviral vectors carrying mouse NMNAT1 gene and RNAi targeting NMNAT1.

METHODSAccording to GenBank, the full-length cDNA sequence of mouse NMNAT1, an interfering sequence targeting NMNAT1 and a negative sequence were designed, synthesized and inserted into plasmid pLenti6 lentiviral vector. The viral stock was prepared by cotransfection of plasmids and the packaging plasmid mix to 293T cells. The virus titer was tested by qPCR methods. After infection of Hela cells with these lentiviruses, the expression of NMNAT1 was detected by qPCR and Western blot.

RESULTSAll the recombinant plasmids were confirmed by sequencing. The titer of virus was over 2 X10(8) TU/mL. Hela cells infected with lentiviral vector carrying full length NMNAT1 gene successfully expressed high-level NMNAT1. The expression of NMNAT1 reduced to less than 30% after delivery of lentiviral vector carrying RNAi sequence.

CONCLUSIONThe lentiviral vectors carrying full length NMNAT1 gene and RNAi sequence targeting NMNAT1 have been successfully constructed.

Animals ; Gene Expression ; Genetic Vectors ; HeLa Cells ; Humans ; Lentivirus ; genetics ; Mice ; Nicotinamide-Nucleotide Adenylyltransferase ; genetics ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

Objective To observe the effects of five flavonoids include rutin(RU),dihydromyricetin(DMY),hesperetin(HP),daidzein(DA)and hydroxysaffor yellow A(HYSA)on myocardiocyte apoptosis induced by H2O2 and to explore their relationships with Keshan disease and the possible mechanism.Methods Primary cultured cardiocytes of neonatal rats were randomly divided into control group,model group,and flavonoids preincubation group.The cardiocyte apoptosis was examined by fluorescent staining,the rates of apoptosis were detected by flow cytometry,the expression of Bcl-2 family proteins associated with apoptosis were observed:by Western blot.Results Compared with model group[(24.33±6.51)%],RU[(13.95±3.80)%],DA[(11.82±3.50)%],HYSA[(12.33±3.78)%]could decreased the rate of apoptosis(P＜0.05).The five flavonoids could up-regulate Bcl-2 expression,down-regulate Bax expression,and increase the Bcl-2/Bax ratio[RU(0.989±0.094),DMY(0.931±0.280),HP(0.980±0.095),DA(1.049±0.092),HYSA(1.031±0.039),vs model(0.490±0.046),the difference had statistical significances(P＜0.05)],but the Bcl-xl did not significantly changed(P＞0.05).Conclusions RU,DMY,HP,DA and HYSA have antiapoptotic effects on cardiomyocyte via regulating Bcl-2 and Bax,which gives us a hint in prevention and treatment of Keshan disease.

This study was purpose to investigate the effects of CD147 on the invasiveness of leukemia cells U937. The experiments were divided into 4 groups: control group, LPS group, CD147mAb group and LPS+CD147 mAb group. Cells were treated by lipopolysaccharide (LPS) or anti-CD147 monoclonal antibody, and the expression of CD147 and MMP-2, -9, the invasive potential of the cells in vitro and ex vivo, as well as the invasion of the implanted tumors in SCID mice were analysed by RT-PCR, FCM, gel zymography and invasion test in vitro respectively. The results showed that the expression of CD147 was elevated by the induction of LPS, and the enhanced expression of CD147 on U937 cells increased the production and secretion of MMP-2 and MMP-9 as measured by reverse transcription-PCR and gel zymography. An increased number of LPS-induced cells invading through a reconstituted basement membrane were observed by invasion assays. These responses were down-regulated after blocking CD147 with anti-CD147 antibody. At 30 days after intravenous injection of LPS pretreated U937 cells to SCID mice human U937 cells were found in the bone marrow and lung of the mice, indicating the invasion of the tumor cells. And overexpressions of CD147, MMP-2 and MMP-9 were found in the lung tissue of the mice injected with LPS-treated but not anti-CD147 antibody treated tumor cells. It is concluded that overexpression of CD147 on U937 cells may increase the secretion and activation of MMP-2 and MMP-9 and thus promote the invasiveness of the tumor cells.
Animals ; Antibodies, Monoclonal ; pharmacology ; Basigin ; genetics ; immunology ; metabolism ; Female ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mice ; Mice, Nude ; Mice, SCID ; Neoplasm Invasiveness ; RNA, Messenger ; genetics ; metabolism ; U937 Cells