A new method to revel RFLPs is presented. The human genomic DNAwas purified by saturatedNaCl solution and the pAW101 probe labelled with digoxigenin-dUTP. The relationships of RFLPsand genetic patterns of PGM1 (phosphoglucomutase),EsD (esterase D),GLO1 (glyoxalase)and ACP(acid phosphatase ) between the fillal generation and parental generation were detected in 15 families(among them 11 cases were aborted fetuses). The probability of paternity (w)was caculated accor-ding to Essen - Moller's formula, each w vlua was over 99. 73 %, reached the standard of incladingpaternity. An effective,rapid, and non-toxic RFLPs technique was established, which is easy to man-age in common lab oratories.

AIM: To construct human GPI-B7-1 fusion protein and investigate the therapeutic potentials in the treatment of tumors. METHODS: The chimeric GPI anchored-B7-1 gene was obtained by overlap PCR and inserted into expressing vector pcDNA3.1, named pc3.1/GPI-B7-1. pc3.1/GPI-B7-1was transfected into CHO cells by lipofectamine ~2 000 reagent. The CHO cells, expressing GPI-B7-1 on membranes, were obtained after selecting by G418. That was confirmed by flow cytometry, SDS-PAGE and Western blot. RESULTS: Recombinant vector pc3.1/GPI-B7-1 was successfully constructed and sequence result indicated that it was identical with reference sequence. The protein on transfected CHO cell membrane selected by G418 was confirmed to be GPI-anchored protein by flow cytometry, and GPI-B7-1 approximately 60 kD was conformed by SDS-PAGE and Western blot. CONCLUSION: A large amount of GPI-B7-1 fusion protein was obtained and will be further studied in the treatment of tumors.2? [

AIM: Centrifuge training can improve forward acceleration (+Gz) endurance. This study was to analyzed the gene expression of rat heart affected by centrifuge, and to research the molecular mechanism of improving +Gz endurance by centrifuge training. METHODS: Differential expressed genes between high+Gz endurance (+16Gz) rats, of test group after trained 12 d and control were screened using suppression subtractive hybridization (SSH) and dot blot hybridization. The obtained expressed sequence tags (ESTs)were used as probes to perform RNA slot hybridization with heart total RNA isolated from each gruop of centrifuge training and high+Gz endurance and low+Gz endurance (+12Gz) rats, respectively. The positive ESTs were sequenced and analyzed using BLAST(nr) at NCBI.RESULTS: Three down-regulated ESTs were obtained from heart samples, all of them are new, and their expression levels were decreasing during centrifuge training. CONCLUSION: Centrifuge training can significantly affect the special gene expressions of rat heart, and the expression changes of these genes may be ralated to the mechainism that +Gz endurance can be improved by centrifuge training.

Objective Whether Schwann cells could promote the survival and differentiation of neural stem cells was explored in vitro. Methods Neural stem cells were dissociated and cloned from the hippocampal tissue of newborn rats.Schwann cells were also dissociated and purified simultaneously from the sciatic nerves and brachial plexus nerves of newborn rats.Then Schwann cells and neural stem cells were Co cultured.The expression of nestin of the neural stem cells and the expression of neurofilament(NF) or glial fibrillary acidic protein(GFAP)of the differentiated cells were detected by immunohistochemistry.The morphological changes of neural stem cells were examined with scanning electronic microscope. Results Compared with control group,the number of surviving neural stem cells and differentiated neuron like cells was significantly increased in Co cultured (Schwann cells add neural stem cells)groups.The primary processes of neuron like cells in Co cultured groups were obviously longer than that in control groups.The irregular convex or concave of the body of neural stem cells became plain and smooth in Co cultured group.Co cultured Schwann cells and neural stem cells have grown to touch together in following manner:1.Body touch body;2.Body touch process;3.Process touch process.Conclusion\ Schwann cells can promote the survival and differentiation of co cultured neural stem cells in vitro.

Aim To explore antitumor mechanism of a recombinant vaccinia virus containing the human CEA-cDNA (rV-CEA). Methods C57/BL mice were immunized three times with rV-CEA. Six weeks later, the macrophages(MΦ s)and splenocytes from rV-CEA-immunized donors were transferred to CEA＋ -HePa tnmor-bearing recipients,Meanwhile, the antitumor effects of these donor's MΦ s and splenocytes and that of the recipient's splenocytes were detected in vitro. Results The MΦ s and splenocytes from rV-CEA-immunized donors possessed strong antitumor activity in CEA-positive tumor-bearing recipients. The in vitro antitumor effect of splenocytes from mice inoculated with MΦ s from rV-CEA-immunized donors were markedly stronger than those from W-VV-immunized donors. However,the in vitro antitumor effect of the MΦ s from rV-CEA-immunized donors was the same as those from W-VV-immunized donors. Conclusion It is demonstrated that antitumor activity induced with rV-CEA may be mediated mainly by antigen present cells (the MΦ s), which activated tumor-specific T cells to kill tumor cells.

ObjectiveTo investigate the role of platelet-derived growth factor(PDGF) and its receptor in the development of hypertrophic scar. MethodsThe expression of PDGF and its receptor were detected in biopsy specimens of 9 pieces of normal skin, 7 pieces of granulation tissue of burn wound and 34 pieces of hypertrophic scar by immunohistochemical staining using specific polyclonal antibodies.ResultsPDGF and its receptor markedly increased in granulation tissue and hypertrophic scars, reaching the peak in the hypertrophic scars within 6 months and then decreased after the peak, whereas PDGF and its receptor expressed weakly in only a few normal skin specimens, and the differences were significant(P<0.05).ConclusionsThe increasing expression of PDGF and its receptor may be related to the development of hypertrophic scar.

Damage to organelles plays a significant role in myocardial ischemia/reperfusion injury,which results in the dysfunction of mitochondria and other related organelles.The communication between mitochondria and other organ-elles can also affect the development of myocardial ischemia/reperfusion injury.For instance,the mitochondria-associated endoplasmic reticulum membrane provides a"seamless connection"and regulates the exchange of organelles and metabolites(such as ions,lipids and proteins)between the mitochondria and the endoplasmic reticulum,which subse-quently affects myocardial ischemia/reperfusion injury.However,there is a lack of studies regarding the interaction be-tween mitochondria and related organelles,which is a critical component in triggering myocardial ischemia/reperfusion inju-ry.Therefore,this article describes the role of mitochondrial crosstalk with endoplasmic reticulum,lysosomes and nuclei in myocardial ischemia/reperfusion injury,and aims to provide a theoretical basis for targeting mitochondrial crosstalk with other organelles in the treatment of myocardial ischemia/reperfusion injury.