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 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 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.

[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.

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

Objective To quantitatively study the distance between the left atrial posterior wall and the descending aorta(DLD)in the normal fetal four-chamber view using fetal echocardiography.Methods The distances between the left atrial posterior wall and the descending aorta in end-systole (DLDs) and in end-diastole(DLDd) were measured in a standard apical four-chamber view using fetal echocardiography in 300 normal fetuses.The gestation ages ranged from 20 to 40 weeks.Fetal non-cardiac biometric parameters included femoral length(FL),biparietal diameter(BPD) and gestation age (GA) based on menstrual age. The correlation between DLDs,DLDd and non-cardiac biometric parameters were analyzed.Results In normal fetuses,the DLDs was (0.32±0.08)cm with a range of 0.19 -0.48 cm,the DLDd was (0.43 ± 0.10)cm with a range of 0.25-0.62 cm.The DLDs and DLDd increased with the growth of GA,and were correlated with GA and BPD and FL significantly (r = 0.545,0.556,0.574,0.530,0.563 and 0.576, respectively,all P <0.001).The linear regression equations between DLDs,DLDd and GA,BPD,FL were:Y =0.009GA+0.041 ,Y =0.038BPD+0.028,Y =0.044 FL+0.073,Y =0.01 1 GA+0.083,Y =0.048 BPD+0.063,Y = 0.054 FL + 0.124,respectively.Conclusions In normal fetuses,the DLDs and DLDd increase with the growth of GA,and have good correlation with GA,BPD,and FL respectively.Normal reference ranges of DLDs and DLDd have been provided.These normative data may be a new tool for assessment of fetal heart,especially has potential applications in screening of total anomalous pulmonary venous connection(TPVAC),absent pulmonary valve syndrome(APVS)or other complex congenital heart defects.

Objective To evaluate the preliminary clinical efficacy of bevacizumab for cerebral radiation necrosis (CRN). Methods Nineteen patients with CRN for whom the treatment with steroids and mannitol failed were retrospectively analyzed with a total of 22 lesions. Except for 5 lesions confirmed by pathological evidence, all lesions were confirmed by the following imaging evidence:1. computed tomography (CT)?or magnetic resonance imaging (MRI)?enhanced lesions showed loss of tension and were accompanied by substantial edema;2. CT?or MRI?enhanced lesions had a low perfusion pressure;3. magnetic resonance spectroscopy indicated that the enhanced areas had a decreased choline peak; 4. positron emission tomography showed that the fluorodeoxyglucose uptake was substantially reduced in the enhanced areas. All patients were given 5 mg/ kg bevacizumab at an interval of 14 days for 2?6 cycles. MRI examination was performed in each cycle before treatment, and the enhanced lesions on T1?weighted images ( T1 WI) and edema on T2?weighted images (T2 WI) were compared before and after treatment. The clinical symptoms, Karnofsky Performance Status ( KPS), and adverse reactions in all patients were evaluated. Comparison before and after treatment was performed by paired t test. Results All 19 patients completed the treatment successfully and there were no severe adverse reactions. The clinical symptoms of patients were substantially improved after the second cycle of treatment, and the KPS score increased by 26?? 8 on average. The visible volume of enhanced lesions on MRI T1 WI was significantly reduced by 54?? 8% after treatment (P= 0?? 000), while the visible volume of edema on MRI T2 WI was reduced by 80?? 7% after treatment (P= 0?? 000). The follow?up time ranged from 3 to 12 months with a mean value of 5?? 6 months. Eleven patients kept clinical improvement in CRN, four patients had recurrence, and four patients died from tumor progression. Conclusions Bevacizumab is preliminarily confirmed to substantially improve the clinical symptoms and quality of life in patients with CRN.