In vitro inhibitory effect of CD8+ cells from patients with aplastic anemia on normal CFU-Mk growth is blocked by cimetidine

AIM: To investigate the effect of CD8+ cells from aplastic anemia (AA) patients and its histamine type II (H2) receptors on the growth of normal CFU-Mk. METHODS: The effects of CD8+ cells and/or cimetidine, on normal human CFU-Mk growth were studied by using CFU-Mk assay. RESULTS: The CD8+ cells from the perpheral blood of AA patients significantly suppressed the growth of normal allogeneic CFU-Mk. This inhibitory effect was blocked by cimetidine at concentration of 1.0×10-5 mol/L. 1.0×10-5 mol/L cimitidine alone didn't inhibit the growth of normal CFU-Mk. CONCLUSION: H2 receptor antagonist cimitidine abolishes the suppressive effect of AA patients CD8+ cells on the growth of normal CFU-Mk.     

Establishment and biological characteristics of a new human endometrial carcinoma

AIM: To establish a new human endometial carcinoma cell line for basic and clinical study. METHODS: The minced tumor tissues were inoculated subcutaneously into the back of nude mice. After forming solid tumor, the mice were killed and the transplant was taken out for primery culture. RESULTS: The established cell line was maintained by serial passages, named EAC. EAC cell line grew rapidly,steadily, and it had similar microscopic morphology to adenocarcinoma cell. Chromosome analysis reveal EAC chromosome counts ranging in hyperdiploid. The expression of ER, PR, P53, MDM2 were negative by SP immunohistochemistry. Tumorigenicity of EAC in nude mice was 100%. CONCLUSION: A new human endometrial carcinoma, EAC, is established successfully, which is helpful for the basic and clinical study of endometrial carcinoma.     

Characters of the fibroblast-like cells cultured from the mobilized peripheral blood cells

AIM: To explore the characters of fibroblast-like (F-L) cells cultured from granunocyte clony stimunating factor (G-CSF)-mobilized peripheral blood cell (PBC) harvests. METHODS: The adherent cells in the PBC harvests were cultured for 2 week in the mediums of RPMI-1640/L-DMEM/G-CSF or interleukin-3 (IL-3) plus RPMI-1640, the cultured F-L cells were analyzed by flow cytometry (FC). RESULTS: The adherent non-confluent F-L cells obtained from the four groups were similar in their phenotypes: CD33+, CD11c+, CD64+, CD14+, CD45+, HLA-DR+, CD86+, CD34-, CD38-, CD3-, CD19-, CD56-, CD29-, CD44-, CD105-. The F-L cells are similar to monocytes except CD38- and were distinct from dendritic cells (DC) or mesenchymal stem cells (MSC). CONCLUSION: The cultured F-L cells are macrophages rather than DC or MSC. G-CSF, rhIL-3 enhances their numbers.     

Meconium induces formation of nitrotyrosine and expression of inducible nitric oxide synthase in the rat lung

AIM: To evaluate the contribution of inducible nitric oxide synthase (iNOS) and nitrotyrosine to acute lung injury (ALI) in rats with meconium aspiration. METHODS: 16 health male Sprage-Dawley rats were randomized to control group and meconium group, followed by intratracheally administration of 1 mL/kg saline or 1 mL/kg 20% human newborn meconium suspension. The animals were killed after 24 h of treatment. The measurements included bronchoalveolar lavage fluid (BALF) cell count, pulmonary myoloperoxidase (MPO) activity and nitric oxide (NO) level. Western bloting was used to determine the expression of pulmonary nitrotyrosine-a specific “footprint” of peroxynitrite and iNOS. RESULTS: Compared to control group, the rats in the meconium group had increased BALF cell counts ［(4.04±1.01)×109cells/L vs （0.53±0.19)×109cells/L］, pulmonary MPO activity ［(1.49±0.22）U/g wet lung tissue vs (0.62±0.16) U/g wet lung tissue］, NO level ［(12.77±5.00） mmol/g protein vs （4.89±1.32) mmol/g protein］, increased expression of nitrotyrosine and iNOS (0.46±0.19 and 1.49±0.60 vs 0.15±0.04 and 0.09±0.04, respectively), all P<0.01. CONCLUSIONS: Meconium results in an increase in expression of pulmonary iNOS, leading to over production of NO and nitrotyrosine, which may be of pathogenic importance in the ALI with meconium aspiration.     

Effect of rhSOD on pulmonary nuclear factor-kappa B and MIP-1α expression in meconium-induced acute lung injury

AIM: To evaluate the role and mechanisms of recombinant human superoxide dismutase (rhSOD) in meconium-induced acute lung injury (ALI) by evaluating pulmonary MIP-1α and NF-κB expression. METHODS: 24 health male Sprage-Dawley rats were randomized to 3 groups (8, each group), followed by intratracheal (IT) administration with (1) saline at 1 mL/kg (control group); (2) 20% human newborn meconium suspension at 1 mL/kg, followed by saline at 1 mL/kg (Mec/saline group); (3) 20% human newborn meconium suspension at 1mL/kg, followed by rhSOD at 20 mg/kg (Mec/rhSOD group). The animal was killed 24 h after treatment. The measurements included the bronchoalveolar lavage (BAL) cell count, RT-PCR analysis of pulmonary MIP-1α mRNA expression, Western blotting analysis of pulmonary NF-κB expression. RESULTS: Meconium-induced ALI was characterized by increased BAL cell count, increased expressions of pulmonary MIP-1α mRNA and NF-κB protein ［(4.68±1.40)×109 cells/L vs (0.53±0.19)×109 cells/L, 3.60±0.75 vs 1.56±0.33, 0.72±0.31 vs 0.23±0.12, respectively in control rats, all P<0.01］. IT administration of rhSOD early in the ALI rat significantly decreased meconium-induced BAL cell count ［(3.13±0.77)×109 cells/L vs (4.68±1.40)×109 cells/L in Mec/saline rats, P<0.01］, inhibited the expression of pulmonary MIP-1α mRNA (2.20±0.39 vs 3.60±0.75, in Mec/saline rats, P<0.01) and NF-κB protein (0.44±0.21 vs 0.72±0.31 in Mec/saline rats, P<0.05). CONCLUSION: The early IT administration of rhSOD in ALI rat following meconium aspiration protects lung from inflammatory injury through inhibiting meconium-induced pulmonary MIP-1α mRNA and NF-κB protein expression.     

An experimental study of hypercholesterolemia-induced disorder of Oddi's sphincter

AIM:To investigate the functional changes and histopathological basis of Oddi’s sphincter under hypercholesterolemia. METHODS:Twenty - four New Zealand rabbits were equally divided into be groups ran- domly. Experimental groups Ⅰ and Ⅱ fed with cholesterol - added forage for 4 and 8 weeks respectively before sacri- ficed. Images, functions and histopathological characteristics of Oddi’s sphincters of experimental groups were studied and compared with control group by cholangiography, catheter manometry, and quantitative analysis of nitric oxide synthase. RESULTS: The hasal pressures of proximal low - pressure segment of Oddi’s sphincter of hath experimen- tal groups Ⅰand Ⅱ (20.9±6.1 mmHg, 25 .6±9.1 mmHg, respectively) were higher than those of control group sig- nificantly (11.7±2.8 mmHg, P < 0.01, P < 0.01 ). The hasal pressures of distal high - pressure "valve" segment of both experimental pops (138 .4±45. 5 mmHg and 144. 5 ±40 .4 mmHg, respectively) were significantly higher than those of coned group (69.3±9.8 mmHg, P < 0.01 and P<0.01). The gallbladder volUmes of experimental group Ⅰ and Ⅱ (3.4±1.0 mL and 3.9±1.9 mL) were significantly larger than those of control group (2.0 ±1.9 mL, P < 0.05 and P < 0.01 ). Between groups there were no significant differences in nitric oxide synthase con- tents. Enlargement of the cell volume, wavy distortion of the cell membrane, loosening and disarrangement of myofila- ment, uneven in size and distribution of dense body, angulation of longitudinal axis between dense body and myofila- ment, swelling, congregated at one end of nucleoulus and disappearance of cristae of plasmosomes and concertration of chromosome at nucleolus margin were observed in experimental groups under electron-microscope. CONCLUSION: The diastole function of Oddi’s sphincter could be damaged by hyperecholesterolemia, inducing a functional disorder of Oddi’s sphincter with increased resistance in bile excretion. The result is retertion of bile in the gallbladder.     

口蹄疫病毒整联蛋白受体的研究进展

从组织分布、各毒株利用率、配体结合区介导感染的能力及β亚基胞质区作用等方面论述了4种整联蛋白αvβ1、αvβ3、αvβ6和αvβ8的研究进展，旨在阐明各整联蛋白决定口蹄疫病毒宿主组织嗜性的作用，并为口蹄疫病毒致病机制的研究提供理论依据。     

正交试验法优化普抗合剂制备工艺

以干浸膏得率和黄芩苷提取率为考核指标,采用正交试验法对普抗合剂水提取醇沉淀制备工艺进行考察.普抗合剂最佳制备工艺方案为加水量10倍,提取2次,每次1 h,55%的乙醇沉淀杂质 .该工艺科学合理,适合于大规模工业生产.     

Influence of APP17 peptide on the learning,memory in SAMP10 mi ce and oxidative stress in mouse mitochondria in hippocampal neurons

AIM:To observe the influence of APP17-mer peptide on the learning and memory,and oxidative stress in mitochondria in hipp ocampal neurons of senescence accelerated mouse/prone 10 (SAMP10).METHODS:The 4-month-old SAMP 10s were randomly divided into mod el group and APP17-mer peptide group (the model group+APP17-mer peptide).The se nescence accelerated mouse/resistance1 (SAMR1) were used for normal control.The APP17-mer peptide group was injected with APP17-mer peptide subcutaneously in a dose of 0.34 μg/per mouse,three times a week;while the model group and the control group were injected with saline.After 28 weeks,water maze test was use d to evaluate the learning and memory of these mice.Then hydroxy lamine reactio n and thiobarbituric acid reaction were used to assay the activity of superoxid e dismutase (SOD) and contents of malondialdehyde (MDA) in mitochondria in hippo campal neurons,respectively.RESULTS:(1) The water maze test showed that the full-course swi mming time and numbers of error committed in model group were significantly incr eased as compared with the normal control (P<0.05).The results in APP17-me r peptide treated group were the same as that in normal control group.(2) Compa red with the normal control,the activity of SOD decreased in model group (P <0.01),and the contents of MDA increased (P<0.01).APP17-mer partially re versed the increase in MDA and the decrease in SOD.The content of MDA was lower and the activity of SOD was higher in APP17-mer peptide group than those in mod el group (P<0.05).CONCLUSION:The learning and memory deficit in SAMP10 are observ ed,and the balance of free radical generation/free radical scavenger is impai red.APP17-mer peptide has an antioxidant character and corrects the abnormal ch ange of SOD and MDA,which may be related to its protective effect of learning and memory.     

Effect of NOS on podocytes in hyperlipidemia rats and the protection of simvastatin in podocyte damage

AIM: To investigate the possible effect of hyperlipidemia on golmerular podocytes,the nitric oxide synthase (NOS) availability and the synthesis of NOS in podocyte damage by hyperlipidemia,further to study the protective effect of simvastatin on podocytes.METHODS: 4 groups of Wistar rats were fed high fat diet for 18 weeks.Serum lipid,urinary protein excretion and renal pathological changes were detected.Immunohistochemistry was used to determine the expression of desmin.The expression of nNOS was detected by Western blotting.RESULTS: The level of serum lipid was increased significantly in hyperlipidemic group and treated group after 4 weeks (P<0.01) and was decreased significantly in simvastatin treated group compared with hyperlipidemic group (P<0.01).Podocyte injury was detected under electronic microscopy in hyperlipidemic group after 4 weeks,and the injury became more serious during the lasting time.The expression of desmin was increased in hyperlipidemic group after 4 weeks,and the level was significantly decreased in treated groups (P<0.01).The urinary protein excretion was increased significantly after 6 weeks (P<0.01),and the level was significantly lower in treated groups (P<0.01).The expression of nNOS was significantly decreased in hyperlipidemic and treated groups (P<0.01),and the level significantly decreased in simvastatin group (P<0.01).CONCLUSION: Hyperlipidemia induces podocyte injury.The injury seems to be associated with NO deficiency and decreased renal NOS activity.The injury can be relieved by simvastatin.