BACKGROUND:neurons are post-mitotic cels that are difficult to survive. Isolation, purification and culture of spinal motor neurons are technical difficulties of cel culture technology. OBJECTIVE:To establish the culture system of spinal motor neurons from neonatal rats and to identify and determine the purity of spinal cord neurons. METHODS: Ventral spinal cord tissues from neonatal rats were made into cel suspension that was subjected to density gradient centrifugation and differential adherence folowed by purification culture. Then, the cels on cover plates were identified and classified using immunocytochemical double staining method, and the content of cel components was calculated in combination with fluorescent Hoechst nuclear staining. RESULTS AND CONCLUSION:The cels adhered wel, and the neurons accounted for 85.8%, among which, motor neurons reached 71.6%, astrocytes accounted for 7.8%, cels negative for neurofilament 200 and glial fibrilary acidic protein were 6.4%. These findings indicate that the ventral spinal cord tissues from neonatal rats combined with density gradient centrifugation and differential adherence can develop high-purity spinal motor neurons.

[Objective]To discuss and analyze the effect of the treatment of high-energy Pilon fractures.[Method]Twenty-four patients with R?edi-Allgower type Ⅱ and Ⅲ Pilon fractures(Ⅱ=14,Ⅲ=10) were treated by open reduction and internal fixation.Ten cases were combined with external fixation.Sixteen cases were treated by bone transplantation(spongy bone=10,big pieces of ilium=6).[Result]From June 2003 to December 2007,the restoration of fractures with Burwell-Charnley system was:excellent=16,good=8;the function of ankle joint with Mazur system was evaluated:excellent=12,good=10,fair=2.[Conclusion]In the time of treating Ruedi-Allgower type Ⅱ and Ⅲ Pilon fractures,the bone transplantation with big pieces of ilium is effective to solve the serious bone loss,strengthen the internal fixation and exercise ankle joint earlier.

Objective To calculate the volume and to study the correlated risk factors of hidden blood loss after artificial hip or knee joint replacement. Methods From July to November in 2008, 38patients with age of (65.11±13.51) years old were treated with artificial hip or knee joint replacement, and were divided by age into four groups. Using Gross formula, total blood loss was calculated depending on height, weight and pre- and post-operation hematocrit, and the hidden blood loss was acquired by subtracting the visible blood loss from the total blood loss. The correlation between age,body mass index( BMI),gender and hidden blood blood loss was observed. Differences of hidden blood loss of artificial hip or knee joint replacement were compared. Results The total blood loss of all arthroplasties was ( 1563.52 ± 693.10) ml,and the hidden blood loss was (538.70 ± 529.77) ml,the percentage of hidden blood loss was 33%. The hidden blood loss was inversely related to age (P＜ 0.05 ). There was no correlation between BMI and hidden blood loss (P＞0.05). The percentage of hidden blood loss in total blood loss in woman was significantly higher than that in man. With age and gender paired, the percentage of hidden blood loss in total blood loss in artificial hip joint replacement was significantly higher than that in artificial knee joint replacement (P＜0.05 ). Conclusions Both of age and gender are risk factors influencing hidden blood loss after artificial hip or knee joint replacement. And compared with artificial knee joint replacement, the percentage of hidden blood loss in artificial hip joint replacement is higher.

ObjectiveTo study the characteristic of apoptosis and the expression of inducible nitric oxide synthase(iNOS),as well as relation between them after spinal cord injury (SCI) in rats.Methods84 adult SD rats were randomly divided into three groups,and made into mild,moderate and severe acute SCI models.The rats were killed at 4h,8h,1d,3d,7d,14d and 21d after surgery.The histopathology changes in spinal cord tissue were observed with HE staining microscopic examination.The expression of iNOS was determined by immunohistochemistry and the apoptosis was labeled by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).ResultsThe histopathology changes in spinal cord tissue deteriorated with increasing in injury degree.A little iNOS immunoreactivity (iNOS-IR) was detected in nerve cells,gliocytes,ependymocytes and blood vessel endotheliocytes of normal and vertebra shelf operated controls,increasing at 8 h, and in flood tide in 7 d. The expression of Bax and the rates of TUNEL positive cells were similar with that of iNOS after SCI. There was positive correlation between expression of iNOS and degree of primary SCI ,apoptosis index(r=0.414,P<0.01;r=0.854,P<0.01).Conclusions Expression of iNOS and Bax increase and a large number of TUNEL positive cells present after SCI in rats. There is positive correlation between expression of iNOS and degree of primary SCI,apoptosis index.

Objective To compare the effects of different doses of FTY720 on inhibiting expression of Caspase-3 and neural apoptosisin rats with acute spinal cord injury (SCI), and find out the suitable dose. Methods 200 female SD rats were randomly divided into 5 groupswith 40 in each group. Group A (laminectomy but not contusion) were administered 0.3 ml normal saline by gavage. SCI model was establishedby Allen's WD method at the T9 level of spinal cord in other groups. Group B were administered 0.3 ml normal saline after modeling.Groups C, D and E were administered 0.3 ml FTY720- saline solution of 1, 3 and 5 mg/kg respectively. All the groups were sacrificed at 6 h,12 h, 24 h, 48 h, 72 h (n=8, per each time-point). Caspase-3 expression was detected with streptavidin-peroxidase immunohistochemistry,and neural apoptosis was detected with the TUNEL method. Results Positive Caspase-3 expression and neural apoptosis were not observedin Group A at 6 h. In Groups B、C、D and E, the number of apoptotic cells increased with increased time of acute SCI, peaked at 24 h afterinjury, and then gradually reduced. Caspase-3 expression was at equal pace with neural apoptosis. The difference of the number of apoptoticand Caspase-3 expression cells among all groups were significant, with the order Group B>Group C>Group D>Group A (P<0.05). However,there was no significant difference between Group D and Group E (P>0.05). The number of apoptotic and Caspase-3 expression cells negativelycorrelated with the dose of FTY720 when the dose was less than 3 mg/kg (P<0.05), and there was no relationship when the dose wasmore than 3 mg/kg (P>0.05). Conclusion FTY720 significantly reduces Caspase-3 expression and neural apoptosis in rats with acute SCI.There is a dose-effect relationship between the dose of FTY720 and the Caspase-3 expression and neural apoptosis. It's indicated that 3 mg/kg is the most appropriate dosage.

ObjectiveTo evaluate the efficacy of mini-incisional double-tsuge suture method with 0-0 absorbable polydioxanone-cord (PDS-Ⅱ)in repair of acute achilles tendon rupture.MethodsA total of 34 patients were subjected to acute closed achilles tendon ruptures,including 25 males and 9 females at a mean age of 32 years ( range,20-45 years).Injury causes included sports injuries in 27 patients,falling injuries in six and heavy object impingement injury in one.The time from injury to operation was average 3 days (range,1-6 days).All patients underwent minimally invasive repair with double-tsuge suture method by using PDS-Ⅱ.The ankle joint was fixed with short leg plaster cast at 30° plantar flexion position and the cast was removed six weeks later to take functional exercise.The patients could walk with full weight-bearing 8-10 weeks later and could gradually return to activity 3-4 months later.Results There was one patient with poor incision healing and one patient with reflex sympathetic dystrophy postoperatively.The rest patients had stage Ⅰ incision healing without skin adhesions.No complications such as infection,lower extremity deep venous thrombosis or sural nerve injury occurred postoperatively.All the patients received follow-up of 12-24 months (average 15 months),which showed no complications like tendon rerupture occurred.According to clinical evaluation criterion of Termann,the average score was 92 points (range,76-96 points).The result was excellent in 28 patients,good in five and fair in one,with excellence rate of 97％.ConclusionsSmall incisional double-tsuge suture method achieves low rate of complications and good outcomes for repairing acute achilles tendon rupture and is an ideal tendon surgery approach.

Human has used maggot to treat diseases for thousands of years. In recent years, with abuse of antibiotic and the rising incidence of antibiotic resistance, maggot therapy, as a surgical alternative, is mainly applied to treat chronic infected wounds on account of its low cost, efficacy and safety. Its mechanisms are disinfection, bio-debridement and enhancement of tissue regeneration. Maggot therapy which serves as a kind of biological therapy is promising. However, living maggot therapy could result in inevitable complications, so that we should apply traditional Chinese medicine theory to investigate and develop new delivery method of maggot. The review summarizes the past and present of maggot therapy.
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