In order to improve the reproductive capacity of adenosine A2A receptor gene knockout mice and strengthen their breed conservation working, the reproductive capacity of adenosine A2A receptor gene knockout mice was analyzed based on their reproductive record. The results showed that while no significant differences of average litter size were observed in the generations of Fback cros , F1 and F2 (P>0.05), the average litter size of Fback cros , F1 and F2 were all significantly higher than that of F3 (P<0.01). It was indicated that the reproductive capacity of adenosine A2A receptor gene knockout mice decreased with the increase of generations, but their reproductive capacity can be improved by back cross.

目的 :探讨以激光微孔化脱细胞真皮基质（LPADM）为载体,骨髓间充质干细胞（MSCs）接种复合自体刃厚皮移植重建皮肤功能的可行性。方法:在已制备的LPADM表皮面接种MSCs,构成含有特殊种子细胞的真皮基质材料（MSCs-LPADM）。取健康雄性裸鼠18只,随机分为三组（每组6只）,麻醉后背部深筋膜层以上制作成2.0 cm×2.0 cm的全层皮肤缺损创面,分别移植三种不同真皮基质（MSCs-LPADM、LPADM、MSCs-PADM）,同期复合移植自体刃厚皮片。A组（MSCs-LPADM+刃厚皮片组）:予MSCs微孔化脱细胞真皮基质复合自体刃厚皮移植;B组（LPADM+刃厚皮片组）:予LPADM复合自体刃厚皮移植;C组（MSCs-PADM+刃厚皮片组）:予MSCs未打孔脱细胞真皮基质复合自体刃厚皮移植。观察皮片大体愈合情况,并于术后3、7、14 d切取移植区标本,行大体、组织学及电镜观察。结果:术后3 d组织学观察A、B两组在移植后真皮基质微孔径结构中均可见空腔样血管结构形成,C组真皮基质中未见血管结构形成。术后7 d,A、B两组移植皮片泛红成活,且两组真皮基质微孔径结构中血管结构更加明显,C组移植皮片出现苍白或变暗坏死。术后14 d,组织学观察示A、B两组真皮基质微孔径结构中可见丰富血管形成,复合皮血管化充分且成活良好,C组胶原结构中仅见少量细胞迁入,未见血管形成,皮片完全坏死。透射电镜显示A组新生的无髓神经样结构,在真皮基质的浅层,可见单个皮脂腺样细胞,深层见单个汗腺样细胞,B组、C组均未见神经样结构的增生及类汗腺细胞、类皮脂腺结构。结论:LPADM复合移植给提前种植的外源性MSCs提供较为理想的增殖分化微环境。对重建移植后皮肤功能提供了一种方法或思路。

主要论述了实验动物学在生物医学发展中的重要性,分析了在温州医学院《实验动物学》课程教学中存在的问题,就如何通过提高教师素质、调整教学内容、更新教学手段和完善考核方式等教学改革方案来提高《实验动物学》的教学质量进行了探索,并对该校《实验动物学》课程的教学改革提出了新的要求。

OBJECTIVE:To evaluate the immune protective effect of Sophora japonical water extract on the spleen cells of ~(60)Co-γ ray irradiated injured mice.METHODS:Mice were randomly divided into 5 groups:normal group(constant volume normal saline),irradiation control group(constant volume normal saline),S.japonical water extract high-dose,medium-dose and low-dose groups(0.8,0.4,and 0.2 mL·kg-1).All the mice were irradiated with ~(60)Co-γ ray except normal group,and then S.japonical water extract was given once a day for 10 days.And then nitric oxide(NO)level and spleen index was detected in spleen cell of mice.RESULTS:NO level of S.japonical water extract medium-dose group was significantly lower than irradiation control group(P0.01),and the spleen index of S.japonical water extract medium-doce group and low-dose groups was significantly higher than irradiation control group(P0.01 or P0.05).CONCLUSIONS:S.japonical water extract can reduce the damage of the irradiated spleen cells,and have the immune protective function.

OBJECTIVE:To determine the expression of vascular endothelial growth factor(VEGF),granulocyte colony-stimulating factor(G-CSF) and ultra-structure of bone marrow in mice with acute radiation injury,in order to evaluate the effect of Sphora japonica on the hematopoietic microenvironment and hematopoietic cells of bone marrow.METHODS:40 mice were randomly divided into 5 groups:normal group,irradiation control group,S.japonica high-dose,medium-dose and low-dose groups(0.8 mL·kg-1,0.4 mL·kg-1 and 0.2 mL·kg-1).The expressions of VEGF and G-CSF in bone marrow tissue were determined by immunocytochemistry SABC staining.Transmission electron microscope was used to observe the change of ultra-structure of bone marrow cells.RESULTS:The expression of VEGF in bone marrow tissue in S.japonica high-dose,medium-dose and low-dose groups was significantly higher than in irradiation control group(P0.05).The expression of G-CSF in S.japonica medium-dose group was significantly higher than in irradiation control group(P0.01),but there was no significant difference in S.japonica high-dose and low-dose groups(P0.05).There was no significant different in the expression of VEGF and G-CSF between S.japonica medium-dose group and normal group(P0.05),but the expression of VEGF and G-CSF in S.japonica high-dose and low-dose groups was lower than normal group(P0.05).The ultra-structure of bone marrow showed bone marrow cells arranged regularly and closely,chromatin was uniform,pyknotic nuclear was not found,rough endoplasmic reticulum was abundant but expanded mildly.CONCLUSIONS:S.japonica can improve the hematopoietic microenvironment in mice with acute radiation injury and promote the growth of hematopoietic cells.

Purpose To study the effect of Sophora japonical on the irradiated mice′s level of malondialdehyde(MDA)and vascular endothelial growth factor(VEGF).Methods Forty Kunming mice were randomly divided into normal control group,Sophora japonical group which including three different concentrationgs groups(0.2,0.4,and 0.8 g/kg),and irradiation control group.All the mice were irradiated with 60Coγ ray besides the normal one,then Sophora japonical group was given for 10 days.After that,MDA level of mice′s serum and the expression of VEGF of bone marrow were detected.Results Compared with the normal group,the MDA level of every Sophora japonical group was increased(P0.05),and the level of MDA was significantly decreased(P0.01) compared to irradiation group.The expression of VEGF of every Sophora japonical group was significantly higher than that of the irradiation group(P0.01).There was no significant difference in the expression of VEGF between Sophora japonical 0.4 g/kg group and normal group(P0.05),but the expression of VEGF of 0.2 and 0.8 g/kg group were both lower than that of the normal one(P0.05).Conclusion Sophora japonical can remove free radical internal oxygen and strengthen anti-oxidization and promote hematopoietic cells renewing.

Purpose To study the effect of Sophora japonical on the irradiated mice′s level of malondialdehyde(MDA)and vascular endothelial growth factor(VEGF).Methods Forty Kunming mice were randomly divided into normal control group,Sophora japonical group which including three different concentrationgs groups(0.2,0.4,and 0.8 g/kg),and irradiation control group.All the mice were irradiated with 60Coγ ray besides the normal one,then Sophora japonical group was given for 10 days.After that,MDA level of mice′s serum and the expression of VEGF of bone marrow were detected.Results Compared with the normal group,the MDA level of every Sophora japonical group was increased(P<0.05),and the level of MDA was significantly decreased(P<0.01) compared to irradiation group.The expression of VEGF of every Sophora japonical group was significantly higher than that of the irradiation group(P<0.01).There was no significant difference in the expression of VEGF between Sophora japonical 0.4 g/kg group and normal group(P>0.05),but the expression of VEGF of 0.2 and 0.8 g/kg group were both lower than that of the normal one(P<0.05).Conclusion Sophora japonical can remove free radical internal oxygen and strengthen anti-oxidization and promote hematopoietic cells renewing.

Thirty six SD male rats weighted 80~100 g were randolnly divided into three grougs,and fed on diets as follows:control(Cu 13.15 mg/kg),copper toxic I(Cu 650 mg/kg)and copper toxic Ⅱ(Cu 850 mg/kg)for studying effects of copper toxicity on liver in rats by the methods of experimental pathology for nine weeks for exploring the effect of high diet copper on some liver parameters.The result showed that lesions were appeared in high copper groupsⅠ and Ⅱ when compared those with control group.There were increased the liver of rats coefficien in high copper groupsⅠ and Ⅱ(P0.05).Also,LDH,GOT and liver copper contents in high copper groupsⅠand Ⅱ were much higher in the control group(P0.01).The control group SOD activities were higher in high copper groupⅠand Ⅱ the dietary copper in range of 650~850 mg/kg caused pathological injures in liver of SD rats.

Objective To breed and identify PAX-8 gene knock-out mice. Methods Heterozygote mice were bred and reproduced.Wild type, heterozygote genotype,and homozygote genotype would appear in the offspring of parents. Genome DNA extracted from the mice's tails was subjected to PCR test for genotype identification. Male heterozygote mice were selected to mate with the female heterozygote mice to acquire homozygote baby mice according to Mendel law. Results The breeding and reproducing were successful and more heterozygote genotype mice were reproduced. Conclusions Appropriate methods of breeding, reproducing, and identifying are effective ways to gain PAX-8 gene knock-out mice from heterozygote mice.

目的:探讨槐白皮对急性放射损伤小鼠免疫调节及骨髓造血细胞的修复作用。方法:取50只昆明小鼠,随机分为槐白皮组、照射对照组和正常组。采用MTT法检测小鼠脾淋巴细胞IL-2水平及免疫细胞化学SABC染色法检测骨髓组织中VEGF的表达水平。结果:IL-2水平检测槐白皮0.2 mg.mL-1、0.4 mg.mL-1、0.8 mg.mL-1浓度组与照射对照组比较有显著性差异（P＜0.05）,与正常组比较无显著性差异（P＞0.05）;且槐白皮浓度0.4 mg.mL-1组最强,槐白皮浓度的0.2 mg.mL-1为最弱。槐白皮各浓度组的VEGF表达水平与照射对照组比较有显著性差异（P＜0.05）,槐白皮浓度为0.4 mg.mL-1浓度组与正常组比较无显著性差异（P＞0.05）。结论:槐白皮可调节免疫功能,增强抗辐射能力,对急性放射损伤小鼠骨髓造血细胞的恢复具有促进作用。

Objective:To study the correlation between bone density and mRNA expressions of TGF-β1 and TNF-α in the alveolar bones of experimental diabetic rats.Methods:Twenty SD rats were randomly divided into normal group and diabetic group(diabetic model).After 8 weeks of study,mRNA expressions of TGF-β1 and TNF-α in the alveolar bone were measured by quantitative reverse transcription polymerase chain reaction(RT-PCR) respectively.Bone density and pathology of the alveolar bones were also examined.Results:In diabetic rats group,the alveolar bone density was decreased,and mRNA expressions of TGF-β1 and TNF-α in the alveolar bone were increased greatly.Conclusion:It is suggested that the alveolar bone with diabetes mellitus have a tendency to develop osteoporosis.It may be related with increased expression of TGF-β1 and TNF-α.

内皮素(ET)和一氧化氮(NO)与糖尿病慢性并发症的发生、发展密切相关.我们以往的研究表明银杏叶提取物(extract of Gingko biloba,Egb)可通过抗脂质过氧化作用和降低NO水平而对糖尿病心肌产生保护作用[1].为进一步探讨其作用机制,本文观察了Egb对链脲佐菌素诱导的糖尿病大鼠血浆、心肌组织NO和ET水平的影响.

OBJECTIVETo study the effects of soybean isoflavone (SI) on born metabolism and morphology in animal model of osteoporosis rats.METHODSAll 70 female Sprague-Dawley (SD) rats were randomly divided into 7 groups according to the levels of total cholesterol (TC) in serum: hyper-lipoid group, estrogen group, low-dose SI group, middle-dose SI group, high-dose SI group, sham group and normal control groups. Bilateral ovaries were extirpated except sham and normal control groups. Except the rats in normal control group, the other rats were fed with high fat diet. Body weight was weighted ad unam vice per week. The estrogen, different dose of SI or deionized water were fed with intragastric administration for 12 weeks. Vena caudalis serum were collected after being ovariectomized, administered for 4 w, 8 w and killed. Serum alkaline phosphatase (AKP) activity and bone density were measured etc.RESULTSTo interfere of estrogen and SI might recover AKP enzyme activity after its being ovariectomized. There almost sowed no differences between high dose SI intervention and estrogen on bone density and microstructure. Bone loss due to being ovariectomized was relieved after SI intervention. SI might protect cardiocyte myofilament and mitochondrial ultramicrostructure. There was mirror image in estrogen, high dose SI group resembling the normal control group, and there was obvious damage in hyper-lipoids group.CONCLUSIONThere should be effects of high dose SI on bone metabolism and morphology in animal model of osteoporosis rats. Serum AKP enzyme activity and bone density should have significantly recovered, the serum level of calcium and phosphorus were maintained after high dose intervened but no significant effects for low dose of SI.

Objective:To study the effects of soybean isoflavone (SI) on metabolism of lipids and lipoprotein (a) [Lp(a)] in ovariectomized rats . Methods:Senventy female Sprague- Dawley (SD) rats were randomly divided into 7 groups according to the levels of total cholesterol (TC) in serum: high-lipid (HF) group, estrogen (EG) group, low-dose SI (L-SI) group, middle-dose SI (M-SI) group, high-dose SI (H-SI) group, sham group and normal control (NC) group. One week after bilateral ovaries were extirpated, except sham and NC group, estrogen, different doses of SI or deionized water were fed i.g. for 12 w. Except NC group, the other groups were fed high fat diet. Body weights were weighed every week and blood and heart were collected at the end of experiment. The serum TC, triglyceride(TG), high or low density lipoprotein-cholesterol (HDL-C, LDL-C), Lp (a), and antioxidative enzymes activities were assayed. Results:After SI intervention, the levels of LDL-C in SI groups were significantly lower than in HF group but higher than in NC group. The levels of Lp (a) were also changed, but there was almost no effect on HDL-C. Persistent intervention with SI can reduce TC, TG , and protect cadiocyte’s actin filament andmitochondrial ultramicrostructure from damage as shown in HF group. The ultramicroscopic pictures in EG and H-SI group almost resembled NC group. Conclusion:Persistent intervention of high-dose SI can reduce the levels of LDL-C, TC, TG and protect the myocardiac damages due to high fat diet in ovariectomized rats.

Objective To study the effects of soybean isoflavone(SI) on lipid peroxidation and liver ultrastructure in ovariectomized rats. Methods Seventy female Sprague-Dawley(SD) rats were randomly divided into 7 groups according to the levels of total cholesterol(TC) in serum: hyper-lipoid group,estrogen group,low-dose SI group,middle-dose SI group,high-dose SI group,sham group and normal control group.After bilateral ovaries were extirpated except sham and normal control groups for a week,the estrogen,different doses of SI or deionized water were fed with intragastric administration for 12 weeks.The rat body weight was weighted once per week and blood samples were collected at the time of ovariectomization,the 4th and 8th weeks after administeration at the time of being killed.The serum TC,triglyceride(TG),high or low density lipoprotein-cholesterol(HDL-C,LDL-C),lipoprotein(a) and antioxygen enzyme were assayed. Results LDL-C levels in SI intervention groups were significantly lower than that in hyper-lipoid groups but higher than that in normal control group;the levels of lipoprotein(a) were changed;there was almost no effect on HDL-C in serum.SI could retain integrity and ultrastructure of hepatocyte;there was similar structure in high dose SI group and normal control group.An obvious damage was detected in hyper-lipoid group.Conclusion SI can improve lipoprotein(a) and lipid peroxidation in ovariectomized rats.A continuous intervention with high-SI can reduce LDL-C to normal lever and retain integrity and ultrastructure of hepatocyte.

蝉拟青霉 Paecilomyces cicadidae(Miquel.)Samson即我国传统中药蝉花[1,2],与冬虫夏草的无性型--中国拟青霉为同属真菌[2],两者的化学成分十分相似.

自从1969年日本学者千原首次报道香菇多糖具有抗肿瘤活性以后,真菌多糖引起越来越多的关注,成为一个非常活跃的研究领域.人们研究发现真菌多糖有很多重要生物学活性,而其中以免疫学活性最为重要,并且常作为一种免疫调节剂.

Objective:To study the influence of soybean isoflavone in lipid metabolism and antioxidation for ovariectomized rats.Methods:The 63 female SD rats after ovariectomized 1 week were randomly divided in to hypsi-lipoids(HF),estrogen,low,middle,high dose,sham operation,normal control groups according to the cholesterol level and intragastric administration with different dose of soybean isoflavone for 3 months.Weight rats ad unam vicem per week and measured about cholesterol(TC),triglyceride(TG),high,low density lipoprotein(HDL,LDL-C),lipoprotein a(Lp(a)),ossature,antioxidation each time after 4,8 and 12 weeks of the female SD rats were ovariectomized.Results:The level of serum TC,HDL-C,LDL-C were higher(P<0.01) and the Lp(a) was lower(P<0.01,in H-SI group) after the rates intervented by soybean isoflavone,the level of serum LDL-C were lower in the HF groups than that in normal control groups,but it was higher than Sham groups.The activity of erythrocuprein(SOD)and glutathion peroxidase(GSH-PX)had highed in intervention groups.Conclusion:Quantity sufficient soybean isoflavone could be effect antagonistic action hyperlipoidemia and improve lipid peroxidation state of ovariectomized rats,except in the stress phase.

Objective To study the effects of soybean isoflavone(SI) on antioxidation and bone morphology in ovariectomized rats. Methods Seventy female Sprague-Dawley(SD) rats were randomly divided into 7 groups,according to the levels of total cholesterol in serum: highlipoids group,estrogen group,low dose SI group,middle dose SI group,high dose SI group,sham group and normal control group.A week after bilateral ovaries were excised except in the sham and normal control groups estrogen,different doses of SI or deionized water were fed with intragastric administration for 12 weeks.Body weights were measured every week and vena caudalis serum,innards tissue,femoral bone were collected immediately after the rats were ovariectomized or killed.The activity of alkaline phosphatase(AKP),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px) in serum and bone density were measured respectively,with 63 SD rats being used for the experiment. Results In the intervention groups femoral bone weights were significantly higher than those in both the high-lipoids and the control group.The activity of AKP、GSH-Px was increased after intervention,There was a same efficacy between high dose intervention and estrogen treatment on bone density.SI invention mitigated ovariectomy-induced bone loss.Conclusion The SI has some efficacy in ovariectomized rats antioxidation,activity of bone metabolism enzyme and can reverse bone loss.The decrease in bone density is interrupted under sufficient intervention.In view of phytoestrogen's weak capability of binding with animals' estrogen receptor(ER),the dosage and the long-term effect of SI applied in clinical intervention need further study.

AIM To investigate effect of losartan on angiotensin and nitric oxide system in diabetic rats kidney,and to elucidate the relation between them.METHODS Rats were randomly divided into 3 groups: control group,diabetic model group and losartan(30 mg·kg~(-1)·d~(-1)×8 weeks,ig) group.mRNA expressions of type 2 angiotensinⅡ receptor (AT_2),collagen Ⅳ and inducible nitric oxide synthase((iNOS)) in renal cortex were measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) respectively.In addition,nitric oxide synthase(NOS) activity,the level of angiotensin Ⅱ and NO in renal cortex were also determined.RESULTS In diabetic model group,urine protein excretion,renal angiotensinⅡ level,collagen Ⅳ mRNA and iNOS mRNA expression were increased obviously as compared to control group.Activity of NOS in diabetic model group was increased,too.However,NO level and AT_2 mRNA expression showed a significant decrease as compared to control group.Administration of losartan markedly ameliorated urine protein excretion and collagen Ⅳ mRNA expressions,but increased NOS activity,NO level,AT_2 mRNA expression and iNOS mRNA expression in renal cortex obviously as compared to diabetes group.CONCLUSION Activation of AT_2 is related to the renal protective roles of losartan,and may involve in the up-regulation of iNOS(mRNA) expression in kidney.