Objective To explore the puncture technic and method of the transabdominal L5/S1 lumbar diskectomy (TALD). Methods Two cases of cadaver were dissected and L5/S1 intervertebral disks were explored and punctured from different angles in order to probe the proper puncture angle for the needling. The radiographs of the lumbar and sacral vertabral samples of 30 cases were scrutinized and analysed to find the pancture imaging marks of the bones. When performing barium meal examinations, the lumbar and sacral spine of 200 cases were inspected to figure out the opacification of the rate imaging marks. 68 cases TALD were reviewed. Results The most pivotal process of the TALD is the needle puncture should be in parallel direction with lumbosacral interspace and then be pushed into the intervertebral disk. In order to make sure of the puncture point two sticking points should be fixed, on the abdomen and the another at the front edges of L5/S1 intervertebral disks. The five imaging makes of the lumbar, sacral, spine together with the neighboring L5/S1 intervertebral disks are of important help to find the front edges. Conclusion The correct puncture method is very important for performing TALD.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a very effective way to make tissue engineer bone vascularization.However, because of expensive and short half-life, VEGF cannot maintain effective concentration in blood after injection. To resolve the problem effectively, gene transfection technique is used in this experiment to transfer human VEGF into seed cells-mesenchymal stem cells (MSCs) of tissue engineer bone and to make it secrete VEGF which could vascularize bone.OBJECTIVE: To explore the possibility of human vascular endothelial growth factor 165 (VEGF165) to transfect rabbit MSCs, and establish the experimental foundation of angiogenesis tissue engineering organization and the treatment of ischemic disorders.DESIGN: Observation control trail.SETTING: First Hospital of Jilin University and Institute of Frontier Medical Sciences of Jilin University.MATERIALS: The experiment was conducted in the Key Laboratory (BSL-2) of Frontier Medical Sciences of Jilin University between June 2003 and August 2004. Health New Zealand white rabbits, 4.0-5.0 months old, weighing 2.5-3.5 kg, half male and half female, were provided by Animal Center of Jilin University. The rabbits were handled under asepsis and anesthetized condition,corresponding to the animal ethical standard. Medicine and reagents: Ham F12 culture media (Gibco, U.S), MTT (Sigma, U.S)PLXSNKDRp-VEGF165 and pcDNA 3.0 vectors were prepared in the present laboratory. ELISA detection kit (Jingmei company,Shenzhen), DH5 α, restriction endonucleases Barn H I, Xhol Ⅰ, Hind Ⅲ, EcoR Ⅰ and standard DNA molecule (Promega,U.S) were also used in this study.METHODS: Rabbits' MSCs were separated and cultivated. The pcDNA 3.0-hVEGF165 expression vector was constructed and identified, pcDNA3.0-VEGF165 eukaryotic expression vector was constructed, the vector was used directly to transfect MSCs. The cultural supernatant then was collected and the soluble protein of human VEGF gene expression was analyzed with ELISA method.The proliferation capability of human umbilical vein endothelial cells (HUVEC) stimulated by the supernatant was measured with MTT methods, untreated MSCs and pcDNA3.0 transfected MSCs were used as control groups.MAIN OUTCOME MEASURES: ① Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid pcDNA3.0-VEGF165;② the secretion of human VEGF165 proteins of the transfected MSCs analyzed by ABC-ELISA; ③ MTT method was used to detect the effects of MSCs culture supematant transfected with VEGF165 on HUVEC cells proliferation ability.RESULTS: ①Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid: The constructed plasmid was digested with Hind Ⅲ and XHol Ⅰ, and then two pieces fragments were isolated with agarose gel electrophoresis, which was accordance with expected results. And sequencing results showed that PeDNA3.0-VEGF165 eukaryotic expression vector was successfully constructed. ② ABC-ELISA method: Compared with the control group, concentration of human VEGF protein in the supernatant of the cultured cells increased significantly after the MSCs were transfected with pcDNA3.0-VEGF165 for 24, 48, 72 hours (P＜0.05).③ MTT method was used to detect the effects of MSCs culture supernatant transfected with VEGF165 on HUVEC cells proliferation ability. The results showed MSCs supematant transfected with VEGF165 (2%, 4%,8%, 16%, and 32%) had statistical significance in promoting HUVEC cells proliferation rate compared with the normal control (P＜0.05).CONCLUSION: Human VEGF gene can be successfully transfected into MSCs and expressed effectively.

A device for promoting normal locomotor activity recuperation was made, which was composed by frame, elastic bolt and normal gait mark carpet. The device has the effects of weight relieving, abnormal gait preventing, and safeguards providing. The patient could do gait training by he/her self or assistant self training under the weight reducing and protecting of the device in order to improve the walking ability and the normal gait formation. It is effective for preventing drop foot and leg adduction, and also helpful for the recovery of normal gait ability and the prevention of abnormal gait formation. It is applicable for the patient who can not be trained with ordinary weight relief walking training, such as severe cerebral palsy and severe spasmodic lower extremity and foot drop after brain injury. The results demonstrated that the device is effective in protecting, correcting, preventing abnormal gait as well as forming normal gait.

Objective To develop one kind of device that can not only reduce the body weight,but also provide a good condition for training lower extremity potentialities.Methods The body weight bearing support and protection system,body weight reducing system and scaled footboard were made respectively.The lower extremity potentialities,nervous conduction speed and leg motion coordination ability training were done under protection and body weight reducing.Results The potential training device could exactly protect the patient and reduce body weight,and make the training step by step.Under the effects of protecting and body weight reducing of the potential training device,the training could be performed together with the Chinese traditional medicine rehabilitation method;that was physical and breathing exercises.The lower extremity potentialities were developed nicely.Conclusion The lower extremity potentialities training device is a useful training device which can bear body weight,have protective effect,develop lower extremity potentialities and improve coordination ability of lower extremities.

Objective To investigate the effect of the eye-acupuncture therapy on serum TNF-α levels in rats with cerebral ischemia-reperfusion injury and the related mechanism. Methods Healthy SD rats were randomly divided into four groups: normal group, sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group according to body weight. Sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group were divided into the 3h group, the 24h group and the 72h group, a total of 10 groups (n=8). To establish the rat model of cerebral ischemia-reperfusion by suture method in ischemia-reperfusion model group and eye-acupuncture group. Eye-acupuncture was separately started immediately after reperfusion and at 30 min before sampling in the 3h eye-acupuncture group, besides, eye-acupuncture was separately taken every 12h in the 24h eye-acupuncture group and in the 72h eye-acupuncture group. The rats in normal group were not treated, those in sham-operated control group were inserted the fishing thread 1cm, and the others were identical with those in ischemia-reperfusion model group. At 3h, 24h, 72h after reperfusion, the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks in ischemia-reperfusion model group and eye-acupuncture group. The method of ELISA was taken to detect the change of serum TNF-α levels in rats after the eye-acupuncture therapy. Results Compared with the ischemia-reperfusion model group, the neurologic impairment score of the eye-acupuncture group decreased obviously; The levels of serum TNF-αin ischemia-reperfusion model group at 3 h, 24 h, 72 h respectively were(76.803±18.325)pg/ml、(85.511±13.334)pg/ml、(86.831±9.232)pg/ml. The level of serum TNF-α in normal group was(24.304±6.511)pg/ml. The level of serum TNF-α in Sham-operated control group at 3 h, 24 h, 72 h respectively were(24.928±3.792)pg/ml,(27.533±5.362)pg/ml,(29.366±5.874)pg/ml. Ischemia-reperfusion model group compare with normal group and sham-operated control group, the difference were significant(P＜0.01). The levels of serum TNF-α in rats after the eye-acupuncture therapy at 3 h, 24 h, 72 h respectively were(40.185±3.335)pg/ml, (48.523±7.687)pg/ml, (51.611± 6.403)pg/ml. Compared with ischemia-reperfusion model group, the levels of serum TNF-α were strongly reduced(P＜0.01). Conclusion The eye-acupuncture therapy could play a role in improving cerebral ischemia-reperfusion injury evidently and the mechanism was related to its reducing serum TNF-α levels.

Objective To evaluate efficiency of brain metastases tumor using X-knife under farctionational stereotactie radiotherapy (FSRT) combine with whole brain radiotherapy (WBRT). Methods Retrospective comparing 51 patients treated by FSRT plus WBRT (FSRT + WBRT group) with 35 patients treated by WBRT alone (WBRT group) on the effecting rate and survival rate. Results The completeness response rate was 49 % and 26 % (P < 0.05) in FSRT + WBRT and WBRT groups, respectively. The effecting rate was 80 % and 71 % (P 0.05) in FSRT + WBRT and WBRT groups, respectively. The middle survival time was (11.0 ± 1.5) months and (6.5 ± 0.5) months (P < 0.05) in FSRT + WBRT and WBRT groups, respeetivley. The 0.5-, 1.0- and 1.5-years survival rate was 63 % and 41 % (P 0.05), 51 % and 23 % (P 0.05) and 24 % and 9 % (P < 0.05) in FSRT + WBRT and WBRT groups, respectively. Conclusions The method with FSRT plus WBRT in the treatment of brain metastases tumor is safe and relieved focal symptom of patients quickly with lesser injury on normal tissue and the survival time be prolonged, it has better therapeutic effects than WBRT alone for treating brain metastases tumor.

Objective To investigate the protective role of human serum albumin in treatment of monocyte-inducible C-type lectin (Mincle)-associated neuroinflammation in subarachnoid hemorrhage (SAH) rats and its underlying mechanisms.Methods Vascular perforation model was used to induce SAH.Ninety-two male SD rats were randomly assigned to sham (n =23),vehicle (n =23),low-dose albumin (0.63 g/kg,n =23) and high-dose albumin (1.25 g/kg,n =23) groups.Saline and albumin were intravenously injected into rats two hours after surgery.Modified Garcia scale was employed to assess neurological functions.Iba-1 and myeloperoxidase (MPO) staining was used to examine the activation of microglial cells and infiltration of neutrophils.Real-time PCR was applied to determine the changes of IL-1β,inducible nitric oxide synthase,CD11b,monocyte chemoattractant protein-1,cytokine-induced neutrophil chemoattractant-1 and CXC motif chemokine ligand-2 mRNA levels.Co-immunoprecipitation was conducted to assess the binding ability of albumin with Mincle.Immunoblotting was carried out to evaluate protein levels of Minlce,Syk and p-Syk.SAH severity measurement was performed before conductions of all the experiments.Results SAH severity scores were 11.4 ± 1.6,12.8 ±2.5 and 11.2 ±3.2 in the vehicle,low-dose albumin and high-dose albumin groups,respectively,without statistically significant difference among groups (F =0.694,P =0.516).Neurological score was 7.5 ± 2.9 in the vehicle group,while the low-dose albumin (14.6 ± 2.2) and high-dose albumin groups (13.6 ± 2.7) exhibited better neurological perfomance (P ＜ 0.01).Immunostaining showed that albumin significantly inhibited the activation of microglia,and reduced the percentage of MPO positive cells from 20.7％ ± 1.9％ in the vehicle group to 12.1％ ±2.1％ in the low-dose albumin group and 9.8％ ±0.9％ in the high-dose albunin group (F =32.216,P =0.001).mRNA levels of pro-inflammatory cytokines and chemotactic factors were also suppressed by albumin (P ＜ 0.05).The results of co-immunoprecipitation displayed that albumin could directly bind Mincle and disrupt the association between Mincle and SAP130.Immunoblotting demonstrated that albumin depressed the protein levels of Mincle,Syk and p-Syk.Conclusion Human serum albumin can inhibit Mincle/Syk-induced neuroinflammation via directly binding Mincle receptor in SAH rats.

Objective To study the expression and significance of cyclin A1 in the skin of wild-type mice at RNA level.Methods Thirty 6-12-week old wild-type Kunming mice(15 male and 15 female)were included in this study.In situ hybridization was used to detect the expression of cyclin A1 mRNA in the skin of head and neck of the mice.The skin treated without probe was regarded as negative control and the testis of male mice was taken as positive contr01.Results The expression of cyclin A1 mRNA was found in sebaceous glands of 25 mice and in epidermis of 12 mice.Strong positive staining of sebaceous glands was seen in 50% and positive staining in 33.3% of sections,whereas strong positive and positive staining of epidermis was seen in 13.3% and 20% of sections.respectively.The positive rate of sebaceous glands was 83.3%,much higher than that of epidermis(33.3%).Conclusions There is a quite high expression of eyelin A1 mRNA in the skin sebaceous gland and epidermis of head and neck of wild-type mice.Especially a strong expression is in the sebaceous glands.It indicates that cyclin A1 mRNA may play a certain role in the physiological function of sebaceous glands and epidermis of the skin.

Objective: To explore the relationship between expression of tumor necrosis factor-α( TNF-α) and electrophysiological heterogeneity in isolated heart tissues and isolated rat ventricular myocytes.The arrhythmogenic mechanisms of TNF-αwere further studied.Methods:Langendorff perfused heart tissues models were used to verify the arrhythmogenic effects of TNF-α.The monophasic action potentials( MAPs) of the endocardium and epicardium from the isolated heart tissues were recorded by elec-trophysiological experiments.The isolated rat ventricular myocytes were obtained by enzymatic dissociation.K+currents(Ito,IK1)were recorded by using whole cell patch clamp technique.Results: Compared to the control group, the difference in MAPD between endocardium and epicardium dramatically increased with TNF-α( P<0.05 ) .TNF-αcould cause MAP duration ( MAPD ) prolongation, and a single dose of TNF-αdifferentially affected the MAPs of endocardium and epicardium of isolated heart tissues.Compared to the control group,the K+currents(Ito,IK1)were dose-dependently decreased with TNF-αin rat ventricular myocytes(P<0.05).However, etanercept had no effects on the MAPD in the absence of TNF-α.Conclusion:TNF-α-induced heterogeneity of MAPD between the endo-cardium and epicardium may provide the substrate for the onset of ventricular arrhythmias during acute myocardial infarction.The effect might be associated with TNF-αcontribute to re-entrant ventricular arrhythmias which resulted from decreased K+currents(Ito,IK1).

Objective To investigate the risk factors for early neurological deterioration (END) in patients with acute middle cerebral artery infarction.Methods From January 2009 to December 2012,81 patients with acute middle cerebral artery infarction completed cerebral angiography admitted to the Department of Neurology,Nanjing General Hospital of Nanjing Military Command were enrolled retrospectively.END was defined as that the National Institutes of Health Stroke Scale (NIHSS) score increased ≥2 or the motor score increased ≥ 1 with in 72 h after admission compared with the baseline score on admission.All the patients were divided into either an END group (26 cases) or a non-END group (55 cases) according to whether the occurrence of END.Univariate factor analysis was used to analyze the differences of the clinical data between the two groups.The grade standard of collateral circulation was assessment with the collateral circulation assessment system of the American Society of Interventional and Therapeutic Neuroradiology/Sociey of Interventional Radiology.Multivariable Logistic regression analysis was used to analyze the risk factors for END after acute middle cerebral artery infarction.Results Compared with the patients in the non-END group,the proportions of age 60 years (65.4％ [17/26] vs.36.4％ [20/55];x2 =5.992,P =0.014),high-sensitivity C-reactive protein level ≥4.0 mg/L (76.9％ [20/26] vs.45.5％ [25/55];x2 =7.080,P =0.008) and diabetes (38.5％ [10/26] vs.16.4％ [9/55],x2 =4.802,P =0.028) in the END group were increased significantly,while the collateral circulation grade was decreased significantly (Z =-3.253,P ＜ 0.01).Multivariable Logistic regression analysis showed that the age ≥60 years (OR,3.412,95 ％ CI 1.075-10.824;P =0.037),high-sensitivity C-reactive protein level ≥ 4.0 mg/L (OR,3.812,95％ CI 1.141-12.740;P =0.030),and collateral circulation grade (OR,2.165,95％ CI 1.241-5.514;P =0.009) were the independent risk factor for END in acute middle cerebral artery infarction.Conclusion The decreased collateral circulation level,age ≥ 60 years and high-sensitivity C-reactive protein≥4.0 mg/L were the independent risk factors for occurring END in acute middle cerebral artery infarction.