ABSTRACT
Objective To evaluate a new method for the isolation of rat pancreatic stellate cells (PSCs) and to investigate the influence of adipose derived stem cells (ADSCs) on PSCs in vitro.Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation.The coculture system of ADSCs and PSCs was set up by transwell insert.The proliferation of PSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of PSCs were tested by Western blot.The apoptosis of HSCs was tested by flow cytometer.The cytokines in the culture solution were tested by ELISA kit.Results The quantity of PSCs was above 5 × 106 cells per rat.The purity and the viability of the cells were about 90-97 percent.After coculture for 72 h,the proliferation and activation of PSCs was inhibited by ADSCs (F =223.27,P ＜ 0.05 ; F =52.97,P ＜ 0.05) and the apoptosis of PSCs was promoted by ADSCs (F =43.62,P ＜ 0.05).more NGF and less TGF-β1 was secreted by ADSCs than by PSCs (NGF:14.68 ± 0.94 vs.8.31 ±0.86,t =4.67,P ＜0.05;TGF-β1:10.65 ±0.46 vs.70.47 ±0.99,t =21.72,P＜0.01).Conclusions ADSCs inhibit the proliferation and activation of PSCs by ADSCs secreting cytokines.
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Objective To evaluate tubal pateney by hysterosalpingography after treatment of tubal pregnancy.Methods 80 patients with tubal pregnancy underwent hysteresalpingography after clinical treatment.Of them,30 were treated with methotrexate of 50mg/m2 intramuscularly( study group,n=30) and 50 were followed up expeetandy (control group,n=50).Results Ipsilateral and contralateral tubal pateneies in the study group were 84% and 97% ,respectively,whereas in the control group were 78% and 92% ,respeetively.There were no statistically significant differences between the two groups.Conclusion Appropriate options for unruptured tubal pregnancy includes either expectant management or methotrexate treatment,both of which result in similar tubal patency rate.
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Objective To prepare polyclonal antibodies of anti Nogo-66, the extracellular region of one central nervous system neurite regeneration inhibitor Nogo, which could be used to further identification and functional study of Nogo molecule.Methods Preparing rabbit anti rat Nogo-66 polyclonal antibodies with a purified Nogo-66 fusion protein expressed in E.coli system. Studying its specificity by Western-blot and immuno-histochemical techniques and identifying its biological activity in PC12 cells.Results The high titer (1∶[KG-*2]10 000) anti rat Nogo-66 polyclonal antibodies were obtained.This antibody could specifically recognize the Nogo protein expressed in E.coli system.Immuno-histochemical staining indicated that the Nogo was widely expressed in rat spinal cord neurons and oligodendrocytes.It could effectively block the neurite extensioninhibition of Nogo protein in PC12.Conclusion Successful preparation of anti rat Nogo polyclonal antibodies provides a useful tool in identification or further functional study of Nogo molecule.
ABSTRACT
The second exons of HLA-DRB genes of Chinese homozygous cell lines were amplified with polymerase chain reaction (PCR), followed by digestion of the amplified DNA segments with the allele-specific restriction endonucleases Cfo 1 and Hinf I. The resulted patterns of restriction fragment length polymorphism (PCR-RF LP) in polyacrylamide gel electrophoresis were used for HLA-DR genotyping. With advantages such as short time-consuming, accuracy and no usage of radioisotope to label oligonucleotide probes for hybridization, the technique has been proved to be capable of subtyping five Chinese HLA-DR 5 cell lines to DRw11 and DRw12 when compared with the PCR-RFLP patterns of reference cell lines. In addition, three Chinese DRw12 cell lines Showed their differences with the reference DRw12 cell BM16 in the genotyping, suggesting that the Chinese DRw12 cell lines might be categorized as new DR5 subtypes or variants related to DRw12.