Twenty-eight adult male rats were used for this study. The experimentar were injected intramuscular with 25 mg/d cortisone acetate and were killed in various intervals separately. The neurohypophysis were fixed in mixed solution of 4% paraformaldehyde and 2% glutaraldehyde in buffered cacodylate. Frozens ections were incubated in the medium of sodium glycero-phosphate and lead nitrate, postfixed in caulfield solution, dehydrated, embedded, ultrathin sectioned, stained With uranium acetate without lead citrate, and observed under EM, In the neurohypophysis of control animals, the active areas of acid phosphatase were few and were limited in lysosome and Golgi apparatus of pituicytes and endothelia of capillaries. After injection of cortisone, the lysosome of pituicytes were increased and the activity of acid phosphatase were strengthened. Besides pituicytes and endothelia cells, there were so many positive thin granules in the processes of dark pituicytes, some areas of positive reaction of AcP appeared in neurosecretion fibers. After increasing the drug dosage, positive reaction of AcP areas showed a tendency of expansion. In this study, we discussed the effect of cortisone on the AcP of neurohypophysis and the functional significance of pituicytes in neurosecretion procedure.

To observe the growth, expansion and differentiation of the cultured bone marrow stroma cell (BMSC) from Macaca Lrus, BMSC isolated from adult Macaca Lrus were cultured with the culture medium confected by ourselves and were induced with some cytokines such as LIF and bFGF. The results showed that the BMSC could proliferate and generate Nestin positive clones when they were cultured in vitro. After subculture, these cells could grow rapidly and differentiated into neuron like cells and astrocyte like cells further, which expressed GFAP or NSE antigen respectively. Therefore, these BMSC possess renewal and differentiation abilities. On the other hand, the culture method we used in this experiment is suitable for culture of BMSC in vitro. The BMSC might be used as the seed cells of the neural stem cells.

BACKGROUND: Researches indicated that bone marrow stem cells (BMSCs) could differentiated into neural stem cells in vitro, but what was the role of neural stem cells(NSCs) in the recovery of cortical injury,whether the NSC is capable of growing and migration in injured still remained unknown.OBJECTIVE: To explore the growing state of autograft NSC derived from crab-eating macaque BMSC transplanted in brain.DESIGN: Prospective case control study based on experimental animals.SETTING: Department of neurosurgery in a hospital of a military medical university.MATERIALS: This study was carried out at Center Laboratory of Neurological Research Institute, Zhujiang Hospital affiliated to the First Military Medical University of Chinese PLA. Six healthy adult crab-eating macaques were purchased from the South China Primate Animal Center.INTERVENTIONS: BMSCs harvested from six crab-eating macaques were cultured in vitro and induced to differentiate into neural stem cells, which then labeled by bromodeoxyuridine(BrdU) and autografted into brains.MAIN OUTCOME MEASURES: Brain tissues underwent hematoxylin and eosin(HE) staining and immunohistochemical staining before observed under optical microscope.RESULTS: The results of HE staining showed that the cell number in injured brain vas obviously higher in both instant and delayed transplanting groups than sham-transplanting group; moreover cells were proved reacting to BrdU by immunohistochemical staining in cortical injuries of both groups at 1-6 months following stem cells autograft, as well as at neighboring white matters at half year later, but no BrdU positive cells could be found in traumatic controls, sham-transplanting group and normal brains.CONCLUSION: NSCs derived from in vitro cultured BMSCs were proved capable of surviving, proliferating, differentiating and migrating in cortex after autograft, so that BMSCs is considered as replacing cells or the source of NSCs; moreover autograft stem cells could survive, proliferate and migrate in old cortical traumatic focus.

Objective:To investigate the regulation mechanisms of the bone marrow mesenchymal stem cells on lymphocyte pro -liferation of type I diabetic rats .Methods:The rat bone marrow mesenchymal stem cells were isolated , cultured and identified and the effect on lymphocyte proliferation of type Ⅰdiabetic rat was observed by MTT assay , and analyze the CD 4 +CD25 +regulatory T cell ra-tio, cell cycle and apoptosis of type I diabetes rat by flow cytometric .Results:B and C groups was significantly lower than the absor-bance values of group A,the differences between the data were statistically significant (P<0.05), C group was significantly lower than group B absorbance values, the difference was significant (P<0.05);the CD4 +CD25 +regulatory T cells of B and C groups were sig-nificantly higher than group A, the differences of the data were statistically significant (P<0.05), the CD4 +CD25 +regulatory T cell ratio of C group significantly higher than that group B , the differences were statistically significant (P<0.05);the apoptosis levels of B and C groups were significantly higher than group A , the differences were statistically significant (P<0.05), the apoptosis levels of C group were significantly higher in group B , the differences were statistically significant (P<0.05).Conclusion:Bone marrow mesen-chymal stem cells can significantly inhibit lymphocyte proliferation of type Ⅰdiabetic rats, and it may regulate CD4 +CD25 +regulatory T cells, promote apoptosis, thereby affecting the immune function of T lymphocytes , and play its rejection.

Objective To explore the effect of fibroblast growth factor receptor(FGFR)inhibitor BGJ398 on the vasculogenic mimicry(VM)formation of glioma cells. Methods The phosphor-FGFR(pFGFR)was de-tected by Western blot,the expressions of MMP2 and MMP14 were detected by Western blot and immunocytochem-istry;the VM formation of U87MG and U251MG was tested by tube formation assay;subcutaneously implanted tu-mor model in nude mouse was established and tumor sections were CD34/PAS double-stained to detect the forma-tion of VM in vivo. Results Western blot showed that pFGFR in the experimental groups decreased significantly (P < 0.05);western blot and immunohistochemical staining showed that the expression of MMP2 and MMP14 in the experimental groups decreased significantly compared to the control group. In the tube formation assay ,the tube formation of U87MG and U251MG cells were restrained. In the subcutaneously implanted tumor model ,the VM number of the experimental group(13.85 ± 3.96)was significantly lower than that in the control group(26.40 ± 5.06,P < 0.05). Conclusions In vivo and in vitro experiments confirmed that BGJ398 can inhibit the activa-tion of FGFR,and inhibit the VM formation of glioma cells. These indicate FGFR signaling pathway is involved in the formation of VM.

Objective To investigate the distribution of the CTX-M- extended spectrum beta-lactamase (ESBLs) producing Esche- richia coli(ECO) and the molecular mechanism of dissemination. Methods To analyze the drug resistance of the 43 isolates, Kirby-Bauer susceptibility method was used. Multiple polymeraso chain reaction (PCR) was used to amplify the gene of ESBLs, AmpC, full length of blaCTX-M-like gene, insertion sequence (IS) ISEcp1B, IS903 , IS26 and integron I. NEST-PCR was used to detect if the beta-lactamase gene lo- cated in the integron I. The product of full length of bla-CTX-M like gone amplified by PCR was sequenced. Results Susceptibility test showed the resistance from high to low in turn was Ampicillin (97.68%), Coftriaxone (67.44 % ), piperacillin(65.12 % ), Cefotaxime (62.79 % ) ,Coftasidime(58.14% ), Cofasolin(55.81% ), Cofepime (53.49%), Cefexitin(51.16%), ciprofloxacin (44. 19% ), Aztreo- nam(41.86% ), Cefoperasone/Sulbactam ( 20.93% ), Amikacin (0% ), Imipenem (0% ), respectively. ECO was susceptive to Imipenem. CTX-M-G1 was found in 25 strains of ECO , TEM, SHV, CTX-M-G1, ISEcp1B, and integron I were found in the nine isolates. IS903 were found in ECO 3 and 5, and IS26 was found in ECO 3. In ECO 3 and 5, blaCTX-M-like was flanked upstream by ISEcp1B element that provided -35 and -10 promoter sequences and a right inverted repeat (IRR) recognized by transposase, downstream by IS903 provided an inverted re- peat, ISEcp1 B and IS903 composed the complex transpeson. Conclusion ISEcplB may drive the expression and dissemination of blaCTX-M-like gene at a high level.

Objective To investigate the clinical effects of surgical treatment of lower lumbar fracture with mini-incision via retroperitoneal anterior approach. Methods The data of 21 cases with serious lower lumbar burst fracture were analyzed retrospectively. There were 15 males and 6 females with an average age of 35.9 years (ranged, 19-65 years). The lesion was located in L3 in 10 cases, L4 in 8 and L5 in 3cases. According to Magerl classification, there was type A (burst in 12 cases, type B (distraction) in 2 and type C (retortion) in 7 cases. All the 21 cases were treated with anterior decompression, bone graft fusion and internal fixation with mini-incision via retroperitoneal anterior approach. The changes of radiograph and neurologic status were recorded respectively. Results All the cases had been followed up for an average of 41.9 months (12-86 months). The radiograph showed obvious improvement on the injured body height (from 42.62％ preoperatively to 94.33％ postoperatively, P＜0.01) and the canal encroachment index (from 2.67 preoperatively to 0.14 postoperatively, P＜0.01). Significant improvement in neurological function were achieved in all patients with the improvement of one grade except for 1 case with L3, T11 fracture and complete paraplegia. No failure of implants was found during the follow-up period. Conclusion Anterior decompression and internal fixation with mini-incision via retroperitoneal anterior approach are successful in treating serious lower lumbar burst fractures.

BACKGROUND: Bone marrow mesenchymal cells, multiple-potential non-hematopoiefic stem cells adhering to the wall in vitro culture, can be induced to proliferate and differentiate towards neurons and glia cells.OBJECTIVE: To investigate the growth state of cat bone marrow mesenchymal cells in vitro culture, as well as the capability to differentiate towards neural stem cells.DESIGN: A randomized sampling study.SETTING: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: This study was carried out at the Central Laboratory of the General Military Neurological Research Institute, Zhujiang Hospital, Southern Medical University between January and December 2002. Twenty healthy home-raised cats, aged 1.0 - 2.0 years and weighing 2. 5 - 4.0 kg, male and female in half, were provided by the Animal Center of the First Military Medical University of Chinese PLA.INTERVENTIONS: Bone marrows were randomly aspirated from the left or right hindlimbs in order to separate bone marrow mesenchymal cells, then the bone marrow mesenchymal cells single cell suspension was co-cultured with neural stem cell culture media in vitro so as to induce differentiation to neural stem cells with tretinoin. CK2 type inverted optical microscope(Olympus,Japan) was used to observe the growth of bone marrow mesenchymal cells in vitro culture, as well as 4, 12, 24, 48 hours of induction upon eliminating or not eliminating the wall-adhering cells. Bone marrow mesenchymal cells in stem cell stage were identified under Olympus optical microscope with modified immunohistochemical staining.MAIN OUTCOME MEASURES: The growth state and the immunocytochemical staining of living bone marrow mesenchymal cells exposed to experimental intervention were observed under the Olympus inverted optical microscope.RESULTS: Data from the 20 cats were analyzed without loss. Reversed microscopic observation revealed that cat bone marrow mesenchymal cells becrame larger when cultured in vitro, which were rich in plasmic granules with prominence projecting, adhering to the wall and forming cell clones. These cells were then successively cultured, and imnunohistochemical staining analysis suggested that the passaged bone marrow mesenchymal cells could express neural stem cells-specific antigen Nestin and differentiate towards glia-like cells and neuron-like cells.CONCLUSION: Cat bone marrow mesenchymal cells possess the characteristics of stem cells; they can be amplified into cell clones and induced to express the property of neural glia cells and neuron-like cells under proper condition.

Viral hepatitis is a major liver disease caused by virus infection .Viral hepatitis is popular in China , mainly caused by hepatitis B and hepatitis C viruses .Experimental animal model is a necessary platform for the research on mechanism of viral infection and pathogenicity , for treatment and vaccine development .Up to date, a great progress in the development of viral hepatitis animal models has been achieved in spite of the most of findings are limited to hepatitis B and C.Here, we summarize the recent findings of viral hepatitis animal models , focusing on the tree shrew animal model and its modeling strategy .

Objective To evaluate endoscopically insertion of feeding tube via nose(EIFTN). Method Jejunal feeding tube was placed endoscopically via the nose in all 136 coma patients. Results This procedure was successful in all patients. The procedure took an average time of 5 minutes. In patients with deep coma,the procedure had no influence on HR,MAP, ECG and SaO2; In semicoma patients, HR and R increased during the procedure (t=3.902, P