Objective To investigate the proliferation of neural stem cells by subcutaneous injection of basic fibroblast growth factor(bFGF),epidermal growth factor (EGF)or combined bFGF and EGF respectively after transient focal cerebral ischemia in rats.Methods The right middle cerebral arteries(MCA) of rats were occluded for 2 h to establish transient focal cerebral ischemia model. bFGF, EGF,combined bFGF and EGF or vehicle were administered subcutaneously at 24, 48 and 72 h after MCA occlusion(MCAO).Thereafter,one administration every 3 d. Proliferating neural stem cells were labeled with bromodeoxyuridine (Brdu) immunohistochemically. Brdu-labeled cells were calculated to characterize neural stem cells in SVZ and the dentate gyrus of the hippocampus at 7 d,14 d,21 d after MCAO.Results Brdu-labeled cells were detected in the bilateral SVZ and the dentate gyrus of the hippocampus after MCAO in each group, the number of positive cells decreased progressively. Compared with the control group, the number of Brdu-labeled cells in treatment group increased significantly after treatment( P

Objective To explore the effects of human umbilical cord blood cells(HUCBCs) transplantation to treat experimental autoimmune encephalomyelitis(EAE) rats and the status of transplanted HUCBCs in the brain and spinal cord tissue of EAE rats.Methods The mononuclear cells abstracted from cord blood of infants were cultured in vitro and marked with 5-bromodeoxyuridine(Brdu) for 48 h.EAE rat models were made and the HUCBCs(3?106) were transplanted into the tail vein(transplanted group) 14 d later.The score of neurological function dificit and the number of the demyelinated foci in brain and spinal cord were undertaken at different time point after transplantation.The statue of survival,differentiation and migration of HUCBCs in vivo were determined by immunohistochemical technique,and compared with control group.Results The scores of neurological function dificit at 21 d,28 d post transplantation in transplanted group were much lower than those in the control group(all P

Objective To explore the effect of human neural stem cells(hNSCs) transplantation to treat cerebral ischemic rats and the status of transplanted hNSCs in the ischemic brain tissue of these rats.Methods Human neural stem cells were separated from 10~13 weeks brains of human embryo and were cultured and induced to differentiate. The middle cerebral artery occlusion rat models were made and the human neural stem cells were transplanted through tail vein 1 day later. Neurological Severity Scores (NSS) tests were undertaken in two groups after transplantation. Immunohistochemistry was used to check the differentiation and migration of human neural stem cells in vitro and vivo.Results Neural stem cells from human embryonic brains had been successfully cultured. These cells formed typical neurospheres in suspension, and the majorities expressed nestin. Three weeks later, the neurological function of rats that received transplantation recovered much better than the rats without transplantation, as evidenced by NSS ( P

Objective To explore the effect of human umbilical cord blood cells (HUCBCs) transplantation to treat cerebral ischemia of rats and the status of transplanted HUCBCs in the ischemic brain tissue of these rats. Methods The mononuclearcells abstracted from 60～100 ml of cord blood of full-term babies were cultured in vitro and marked with 5-bromodeoxyuridine (Brdu)(5 ?mol/L) for 2 days. The middle cerebral artery occlusion rat models were made and the HUCBCs (3?106) were transplanted into the lateral ventricular 1 day later. Neurological severity scores (NSS) tests were undertaken at different time point after transplantation, and iimmunohistochemistry method was used to check the migration and differentiation of HUCBCs. Results The HUCBCs had the capacity of proliferation in vitro and were induced to differentiate into astrocytes, oligodendrocytes and neurons in vivo. 3 weeks later, the neurological function of rats that received transplantation recovered much better than the rats without transplantation, as evidenced by NSS (all P

[Objective]To explore the effect of tacrolimus on expression of hepatocyte growth factor(HGF)in rat spinal cord following peripheral nerve injury.[Method]Fifty male rats were randomly divided into normal group,injury group and treatment group.Models of peripheral nerve injury were established by transection of bilateral sciatic nerve at 0.5 cm distal to piriform muscle.Then treatment group received subcutaneous injection of tacrolimus(1 mg/kg)at the back of the neck,while injury group received 0.9 % saline.The L4~6 spinal cords were harvested at various time points after surgery.Western blotting and immunofluorescence staining were used to detect the level and position of HGF in the spinal cord.[Result]HGF-positive neurons were located in anterior horn,intermediate zone and posterior horn of gray matter in normal spinal cord.There was no significant difference in the expressions of HGF between injury group and normal group,while the expression of HGF was significantly higher in treatment group than that in injury group at 7 and 14 days after surgery.[Conclusion]Peripheral nerve injury does not up-regulate the expression of HGF in spinal cord,while tacrolimus can induce high expression of endogenous HGF after injury to protect neurons and promote axonal outgrowth.

Objective To retrospectively evaluate the clinical effects of end-to-end suture in repairing acute closed Achilles tendon ruptures.Methods Twenty-eight consecutive patients(20 males and 8 females,aged from 19 to 48 years with a mean of 36.5) with acute closed Achilles tendon rupture admitted from Nov.2000 to Jun.2006 were involved in present study.All patients were produced to have complete Achilles tendon rupture by MRI,and they received surgical procedure within 4 days after injury.An incision of 1cm long was made medial to the tendon,sparing the lesser saphenous vein and sural nerve.The tendon stumps were minimally debrided and end-to-end sutured using the Kessler or Bunnell techniques.The patients were followed-up for 12 to 36 months(mean 20 months),the therapeutic effects were evaluated by Arner-Lindholm scoring system,and postoperative complications were documented during the follow-up period.Results Of the 28 patients,19 got excellent therapeutic effect and 9 got good as evaluated by Arner-Lindholm scoring system,the satisfactory rate was 100%.All the patients regained normal gait,and rehabilitative training was initiated 3 months after operation.The postoperative complications included superficial wound infection in one case and skin edge necrosis in one case,resulting in a 7.1% incidence of complication.No peripheral nerve injury or Achilles tendon re-rupture occurred during the follow-up period.Conclusions An excellent therapeutic effect and minimal postoperative complication may be expected with end-to-end suture for repairing acute closed Achilles tendon rupture.

To find a new source of seed cells for constructing tissue-engineered intervertebral disc, nucleus pulposus (NP) cells and mesenchymal stem cells (MSCs) were isolated from New Zealand white rabbits. The nucleus pulposus cells population was fluorescence-laelled and co-cultured with MSCs with or without direct contact. Morphological changes were observed every 12 h. Semi-quantitative reverse transcriptase-polymerase chain reaction was performed to assess the expression levels of Sox-9, aggreacan and type II collagen every 24 h after the co-culture. MSCs treated with direct contact rounded up and presented a ring-like appearance. The expression of marker genes was significantly increased when cells were co-cultured with direct contact for 24 h. No significant change was found after coculture without direct contact. Co-culture of NP cells and MSCs with direct contact is a reliable method for generating large amount of NP cells used for cell-based tissue engineering therapy.
Aggrecans/metabolism ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Collagen/metabolism ; Gene Expression ; Gene Expression Regulation ; High Mobility Group Proteins/metabolism ; Intervertebral Disk/*cytology ; Mesenchymal Stem Cells/*cytology ; Mesenchymal Stem Cells/metabolism ; Models, Biological ; Reverse Transcriptase Polymerase Chain Reaction ; SOX9 Transcription Factor ; Tissue Engineering/instrumentation ; Tissue Engineering/*methods ; Transcription Factors/metabolism

This study is to investigate the effect of FK506 on expression of hepatocyte growth factor (HGF) in rats' spinal cord following peripheral nerve injury and to elucidate the mechanisms for neuroprotective property of FK506. Fifty male rats were randomly divided into normal group, injury group and treatment group. Models of peripheral nerve injury were established by bilateral transection of sciatic nerve 0.5 cm distal to piriform muscle. Then the treatment group received subcutaneous injection of FK506 (1 mg/kg) at the back of neck, while the injury group was given 0.9% saline. The L(4-6) spinal cords were harvested at various time points after the surgery. Western blotting and immunofluorescent staining were used to detect the level and position of HGF in spinal cord. Immunofluorescent staining showed that HGF-positive neurons were located in anterior horn, intermediate zone and posterior horn of gray matter in normal spinal cord. Western blotting revealed that there was no significant difference in the expressions of HGF between the injury group and the normal group, while the expression of HGF was significantly higher in the treatment group than in the injury group 7 and 14 days after surgery. It is suggested that peripheral nerve injury does not result in up-regulation of the expression of HGF in spinal cord, while FK506 may induce high expression of endogenous HGF after injury thereby protecting neurons and promoting axonal outgrowth.
Cells, Cultured ; Gene Expression Regulation ; Hepatocyte Growth Factor/metabolism ; Immunosuppressive Agents/metabolism ; Immunosuppressive Agents/*pharmacology ; Microscopy, Fluorescence/methods ; Neurons/metabolism ; Peripheral Nervous System/*metabolism ; Sciatic Nerve/metabolism ; Spinal Cord/*cytology ; Spinal Cord/metabolism ; Spinal Cord Injuries/*drug therapy ; Tacrolimus/metabolism ; Tacrolimus/*pharmacology

Background and purpose:It was reported that erythropoietin may directly or indirectly induce the tumor cells to proliferate and result in diseases progression when recombinant human erythropoictin is used clinically in cancer-related anemia.Recently,the expression of erythropoietin-receptor(Epo-R)was detected in breast cancer.This study was done to detect the expression of Epo-R,estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor 2(Her-2)in breast cancer,and to investigate the relationships between these indexes and the clinical significance.Methods:Sixty breast cancer patients were analyzed,the expression of Epo-R and microvascular density(MVD)were detected by immunohistochernistry,in order to clarify the relationships between the expression level of Epo-R,MVD and ER,PR,Her-2.Results:The rate of Epo-R expression was 78.3%,the mean number of positive tumor cells was 39±24,while the tissue MVD was 25±9.The expression level of Epo-R was positively correlated with the MVD.Also,the level of MVD was higher in the group of Epo-R positive than the negative,which has significant difference(t=3.4252,P＜0.001).The expression level of ER,PR has no definite relationships with Epo-R,MVD.The expression level of Her-2 was both closely associated with Epo-R,MVD.The expression level of Epo-R has an obvious relationship with clinical stage,lymph node status and the size(P＜0.001).The value of MVD was positively correlated with lymph node status,while not with clinical stage and the size.Conclusion:The expression level of Epo-R was markedly higher in breast cancer,and has a positive correlation with the tissue MVD.The expression level of Epo-R and MVD were significantly associated with Her-2,but not with ER and PR.It may contribute to clarifying the clinicopathological characteristics and prognosis of breast cancer by detecting both the expression level of Epo-R and MVD.

Objective To determine the frequency of depression in patients with Parkinson' s disease(PD) and healthy controls over 50 years of age and to analyze the characteristics and influencing factors of PD with depression(PDD) in Nanjing.Methods One hundred and twenty-six PD patients were diagnosed and assessed using Self-rating Depression Scale (SDS) and Hamilton Depression Scale (HAMD).The frequency,characteristics and influencing factors of depression were statistically analyzed,and the factor analysis of HAMD was carried out.Also,one hundred and twenty-four healthy subjects over the age of 50 were selected as the control group.Ressults The incidence of depression in PD group was 48.4％ ( 61/126):15.1％ (19/126) for mild depression,27.8％ (35/126) for moderate depression,5.6％ (7/126) for severe depression.The incidence of depression in the control group was 9.7％ (12/124):5.7％ (7/124) for mild depression,2.4％ (3/124) for moderate depression,1.6％ (2/124) for severe depression.There was a significant difference between these two groups( x2 =45.36,P ＜ 0.01 ).Univariate and Logistic regression analysis revealed that a high frequency of depression occurred in patients with long PD duration,high H-Y stage and UPDRS Ⅲ.According to each factor analysis of HAMD,the scores of cognitive impairment,tardiness,anxiety and sleep disturbances of the PD patients with depressive syndromes were higher than that of the control group.Conclusion Depression is a relatively common complication of PD in Nanjing which is associated with long PD duration,severity of motor disturbances and increasing H-Y stage.