Objective To investigate the effects of a highly selective protein kinase A (PKA) inhibitor, H89, injected intrathecally (IT) on hyperalgesia and phosphorylation of cAMP element binding protein(pCREB) in the dorsal horn neurons of the spinal cord induced by chronic constriction injury (CCI) to the sciatic nerve.Methods Fifty-eight adult female SD rats weighing 230-270 g were used in this study. CCI was produced by 4 loose ligatures place on the sciatic nerve of right hind leg at 1 mm interspace with 3-0 silk suture. The experiment was carried out in two parts. In part Ⅰ 28 animals were randomized to receive H89 1 (group H1), 2 (group H2 ) or 4 nmol (group H4) or 10 ?l of DMSO (the solvent) 10mmol?L-1 (control group) intrathecally (IT) (n = 7 each) 7 days after surgery. The paw-withdrawal latency following mechanical (MWL) and thermal stimulation (TWL) were recorded before (baseline) and 15, 30 and 60 min after drug administration. In part Ⅱ 24 rats were randomized to receive H89 1, 2 or 4 nmol or 10 ?l of DMSO 10 mmol?L-1 IT as in part Ⅰ (group H1, H2, H4 and control group, n = 6 each) . Another 6 animals received 10 ?l of DMSO 10 mmol?L IT 7 days after sham operation. The animals were killed 30 min after drug administration and lumbar (L4.5) segment of spinal cord was removed for determination of pCREB expression in the dorsal horn neurons of spinal cord using immuno-histochemical technique. Results MWL and TWL were significantly increased after drug administration in group H2(at 15min) and group H4(at 15 and 30 min) as compared to the baseline values(P

Objective To determine whether chronic constriction injury (CCI) to sciatic nerve is associated with changes in the phosphorylation of CREB in dorsal root ganglia ( DRG) and superficial dorsal horn neurons of the spinal cord.Methods Thirty-two adult female SD rats weighing 230-270 g were randomly divided into 4 groups (n = 8 each): Ⅰ blank control;Ⅱ sham operation; Ⅲ CCI 2w and Ⅳ CCI 4w. The animals were anesthetized with intraperitoneal pentobarbital 40 mg?kg-1. Right sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm interspace with 3-0 silk suture. Paw withdrawal threshold to mechanical stimulation (von Frey filament) applied to plantar surface ( MWT) and paw withdrawal latency to thermal stimulation (radiant heat) (TWL) were measured before operation (baseline) and 14 days (group Ⅰ,Ⅱ and Ⅲ) or 28 days (group Ⅳ) after nerve ligation. The animals were killed the next day and the L4,5 segment of the spinal cord and L5 dorsal root ganglion were removed for determination of expression of phosphorylated-CREB-immuno-reaction(pCREB-IR) using immuno-histochemistry. The pCREB-IR cells both in DRG and superficial dorsal horn neurons were quantified and analyzed. Results The animals developed mechanical and thermal hyperalgesia on the 14th day after CCI (in group CCI 2w) . The hyperalgesia was greatly attenuated on the 28th day after CCI. Interestingly enough the animals in sham operation group (Ⅱ) also developed mechanical hyperalgesia to some extent on the 14th day after operation. The number of pCREB-IR cells was significantly increased in the ipsilateral L5 DRG and superficial dorsal horn in group Ⅲ(CCI 2w) as compared to sham operation group ( P

Objective To analyze the clinical effect of minimally invasive transforaminal lumbar interbody fusion (MI-TLIF) in combination with unilateral pedicle fixation on lumbar degenerative diseases in the elderly.Methods From June 2011 and June 2013,51 elderly patients with lumbar degenerative diseases who underwent MI-TLIF in combination with unilateral pedicle fixation,including 39 males and 12 females,aged from 60 to 72 years (average 65 years old),were included in this study.All the 51 cases had mono segmental lesion,including 11 cases with the lesion in L3.4,22 cases with the lesion in L4.5,and 18 cases with the lesion in L5-S1.The operation time,blood loss,postoperative drainage and complications were recorded.Postoperative outpatient follow-ups were conducted once every 2-3 months for all cases,and JOA scoring criteria (29 points) of lower back pain were adopted to evaluate the clinical effects.Thin-layer helical CT scanning reconstruction was applied to evaluate the fusion rate of bone grafting.X-ray film of lumbar vertebra AP & LAT was employed to evaluate the intervertebral space height,the internal fixation and the position of cage.Results The operation time ranged between 50-90 min with a mean value of 65 min; blood loss ranged between 80-180 ml (average 110 ml); and postoperative drainage volume ranged between 20-70 ml (average 40 ml).Healing phase I was achieved in 49 cases,while post-operative local cutaneous necrosis around the incision site occurred in 2 case and the wound healed well after wound dressing.Dural laceration without nerve root injury occurred in 2 cases.The follow up in 51 patients ranged between 18 24 months (average 20 months).In terms of JOA scoring,the preoperative score was (9.7± 1.6),the post operative score was (21.4 ± 1.4) 7 d after operation and the score was (25.4 ± 1.2) in the final follow up.During the final follow-up,35 cases were evaluated as excellent,11 cases as good and 5 cases as moderate,the excellent and good rate was 90.2％,and the fusion rate of bone grafting was 88.2％.The intervertebral space height ranged from (9.2±1.5) mm before operation to (11.2±1.3) rnm 7 d after operation,to (11.0±1.2) mm in the final follow-up.There were significant difference in intervertebral space height before and after operation (both P＜0.05),and no difference was found between 7d after operation and the final follow-up (P＞0.05).Looseness,fracture or cage displacement were not observed.Conclusions The clinical effect of MI-TLIF in combination with unilateral pedicle fixation on lumbar degenerative diseases in the elderly is satisfied.It has the advantages of high fusion rate,less trauma,quicker recoverer,higher cost effectiveness and convenience.However,the indications for operation should be restricted and the long-term curative effect remains to be studied in further clinical studies.

Objective To evaluate the effect of electric vagal stimulation on postoperative cognitive dysfunction in aged rats.Methods Thirty healthy Sprague-Dawley rats of both sexes,aged 18-20 months,weighing 390-550 g,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),surgery group (group S),and electric vagal stimulation group (group V).Incision of the right carotid sheath was performed,and the vagal nerve was exposed after anesthesia in S and V groups.The right vagus nerve was stimulated for 30 min with continuous electric rectangular pulses (1 ms,10 Hz,1-2 V) after surgery in group V.The Morris water maze place navigation test was performed at 4,3,2 and 1 days before surgery and 2 days after surgery.The cognitive function was assessed using spatial probe and open field tests conducted at 2 days after surgery.The escape latency,the number of crossing the platform,the number of crossing the grid,the number of standing on the back legs,and the time animals spent in the central square were recorded.After the end of behavioral testing,venous blood samples were collected from the jugular vein for determination of concentrations of serum tumor necrosis factor-alpha (TNF-α),interleukin (IL)-1β and IL-6 using enzyme-linked immunosorbent assay.Results Compared with group C,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly decreased,the escape latency and the time animals spent in the central square were significantly prolonged,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly increased at 2 days after surgery in group S (P＜0.05).Compared with group S,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly increased,the escape latency and the time animals spent in the central square were significantly shortened,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly decreased at 2 days after surgery in group V (P＜0.05).Conclusion Electric vagal stimulation can improve the postoperative cognitive dysfunction in aged rats.

Objective To evaluate the effect of transcatheter hepatic artery embolization with lipiodol on perfused radiofrequency ablation (PRFA) on mini-porcine.Methods Ten Chinese mini-porcine were randomly divided into embolization group and control group (each n=5) .A standard PRFA with infusing cooling procedure was done in control group,while transcatheter hepatic artery embolization with lipiodol was done before PRFA in embolization group.CT and MR scan were performed after PRFA.Then a comparative analysis of the shape and volume of the lesions in the liver were performed.Resuits In both two groups,distinct spherical margin of the PRFA lesions was obtained.The minimal diameter was (31.76±3.43) mm in embolization group and (27.23±3.49) mm in control group (t=2.675,P＜0.05) .The average volume of the embolization group was (54.47±9.98) cm~3,while in the control group was (20.90±5.68) cm~3 (t=10.424,P＜0.05) .Conclusion Transcatheter hepatic artery embolization with lipiodol before PRFA can enlarge the ablation size of mini-porcine's liver.

Objective To investigate the efficacy and safety of radiofrequency ablation (RFA) combined with transarterial chemoembolization (TACE) for treating of hepatic metastasis. Methods From Mar. 2005 to Oct. 2010, 22 males and 14 females with hepatic metastasis were enrolled in this study. Mean age of the patients was 63±12 (42-82) years. Tumor size was (4.5±2.4) cm (min.1.5 cm, max. 12.0 cm). Totally 47 lesions were treated with single metastasis in 29 cases and multiple ones in 7 cases. All cases were failed to chemotherapy or could not stand for the side effect of chemotherapy. Contrast enhanced CT scan was given to all patients before RFA+TACE. For lesions with rich blood supply, TACE was given and then RFA. For those with poor blood supply, RFA was given first and then TACE. For multiple lesions, RFA+TACE was given one by one for each lesion. As for follow up, ultrasound and blood check was given monthly. Enhanced CT scan was given every 3 month. For residual lesions or recurrent lesions, RFA+TACE were given repeatedly. The whole patients was divided into two groups according to the image follow up including complete ablation group and partial ablation group. For complete ablation group, no further treatment was given. For partial ablation group, if it was not suitable for further RFA, repeated TACE was given there after. The end point of follow up was death event. Survival of the whole group and the two subgroups was analyzed statistically by Kaplan-Meier method. Results All RFA procedures was given under intravenous anesthesia and local anesthesia, no severe complication was noted. Lesions in 16 patients were completely ablated after single or multiple sections of RFA+TACE. Twenty patients were in the partially ablated group. Follow up time was 25±10 (10-40) months. Twenty-three patients died and 13 kept alive during the follow up time. The estimated median survival time was 27 month (95%CI: 24-32 months). Survival ration at 1, 2, 3 years for the whole group was 91.7%(33/36),55.5%(20/36),36.1%(13/36) for the whole group. The 3 years survival for complete and partial ablation group was 75.0%(12/16),5.0%(1/20),there was a significant difference between the two groups(P<0.01). Conclusion For patients with hepatic metastasis, RFA+TACE can effectively control the local lesion. Complete ablation is the key point for a better survival.

Objective To investigate the feasibility and efficiency of unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion to treat low lumbar vertebra diseases. Methods Thirty patients with low lumbar vertebra diseases were entered into the study, including 8 males and 22 females with an average age of 53.7 years. All patients underwent discectomy, spinal canal decompression, cage implantation and lumbar fixation by unilateral pedicle screw combined with contralateral translaminar facet screw under gunsight guiding by percutaneous. Clinical outcomes were assed by JOA questionnaires before and after operation. Operative time, blood loss, and postoperative draiming were recorded. Radiological examination was obtained to assess position of translaminar facet screw.Results Mean operation time was 89 min with a blood loss of 285 ml. Position of translaminar facet screw grade Ⅰ were 24 cases, and grade 11 were 6. Mean follow-up was 22.5 months. 29 cases got bony fusion, and the fusion rate was 96.7%. There were no instability and evidence instrument failure during follow-up. The JOA grades improved from 13.0 preoperation to 25.2 at final follow-up, with the excellent and good rate of 72.5 %. Conclusion Unilateral pedicle screw combined with contralateral translaminar facet screw fixation by percutaneous and interbody fusion provide simple procedure, little trauma, forceful fixation, high fusion rate, and less complication, etc. Therefore, the surgical maneuver is a good choice for partial low lumbar vertebra diseases.

UL48 plays essential role in replication of MDV genome and interacts with UL36 as well as other MDV tegument proteins.To investigate the interaction between UL48 and UL36 during MDV oncogenisis,antibody against UL48 was prepared and characterized in current study.UL48 gene was amplified from MDV-Ⅰ genome and then subcloned into pTYB1 and pGEX-4T3 vectors for UL48 expression with induction of IPTG in BL21(DE3) E..coli cells.Chitin-sepharose and Glutathion-sepharose were,respectively,used to purify fusion protein intein-UL48 and GST-UL48.Four subcutaneous injections of intein-UL48 fusion protein were done on the lower back and the thigh of rabbit and then other three injections with an interval 10 days.The titer of antibody was measured by the sandwich ELISA with UL48 protein isolated from GST-UL48 after cleavage of thrombin.Western blot was carried out for specificity analysis of antibody against UL48 protein.The results suggested that UL48 antibody was succesfully prepared,and its titer was 1 ∶ 512 000.

Objective To investigate the feasibility and effectiveness of bilateral pedicle screw fixation combined with in-terbody fusion for the treatment of lumbar spondylolisthesis (degree Ⅱ or Ⅲ) through muscle-splitting approach. Methods There were 52 patients (16 males and 36 females) analyzed in our study from January 2012 to December 2015. The average age was 54.9 ± 7.21 years (from 35 to 70 years). The history of disease was from 10 months to 15 years, with an average of 66.6 ± 32.71 months. Diagnosis included:degenerative spondylolisthesis in 2 cases, and isthmic spondylolysis in 50 cases. The sites included L4,5 in 18 cases and L5S1 in 34 cases. The degree was Ⅱ in 35 cases and Ⅲ in 17 cases. 27 cases were treated by bilateral pedicle screw fixation combined with interbody fusion through median incision approach (open group) and 25 cases through muscle-split-ting approach (minimally invasive group). The clinical and imaging results were observed and compared between the two groups, including operation time, intraoperative blood loss, postoperative incision drainage, VAS score of lumbar incision pain at 72h post-operative, intraoperative and postoperative complications, interbody fusion rate, multifidus muscle areas and grades at 1 year post-operative, and the improvement of ODI index. Results All the cases were operated successfully. The operation time of open group was shorter than minimally invasive group. However, the intraoperative blood loss and postoperative incision drainage of minimally invasive group was statistically less than open group. The VAS scores of postoperative 72 h were 3.25 (2 to 6) in open group and 1.62 (1 to 3) in minimally invasive group. And the difference was statistically significant. The misplacement rate of pedi-cle screws of minimally invasive group was lower than open group. The average follow-up time was 25.5 months (ranged from 12 to 36 months). The multifidus muscle areas and grades of minimally invasive group were superior to open group at 1 year postopera-tively. Compared to preoperative, the slipping angle, slipping rate, sacral inclination angle and vertebral space height were signifi-cant improved after operation in two groups. The ODI index was improved postoperatively in both two groups, and the difference was statistically significant at the final follow-up. The interbody fusion rate was 92.6％in open group and 96％in minimally inva-sive group, and there was no significant difference between the two groups. No cerebrospinal fluid leak, never injury, pedicle frac-ture or wound infection was found after the operation in two groups. There was no pedicle screw loosening, cage shifting, or degen-eration of the adjacent segment observed during follow-up. Conclusion Bilateral pedicle screw fixation combined with inter-body fusion through muscle-splitting approach has advantages of small incision, less trauma, less bleeding, quick recovery and well clinical results, which can be a better choice in treating lumbar spondylolisthesis (degree Ⅱ or Ⅲ).

Objective To evaluate the effectiveness and radiographic outcomes of percutaneous kyphoplasty (PKP)/percutaneous vertebroplasty (PKP) in the treatment of stage Ⅲ Kümmell's disease without nerve injury.Methods From September 2012 to September 2016,there were 28 patients with stage Ⅲ Kümmell's disease recruited to study,of which 16 patients were treated with PKP and the other 12 cases with PVP.with a mean age of 78,ranging from 67 to 95,consisted of 3 males and 29 females.All patients had single vertebral body lesions without symptom of spinal cord injury.Visual analogue scores (VAS) and oswestry dysfunction index (ODI) were recorded before surgery,on the 7 d after surgery and at the last follow-up.Meanwhile,Cobb angles,anterior and posterior heights of the injured vertebral body were measured by X-ray lateral radiography at these three time points.Furthermore,the condition of bone cement in the injured vertebral body was observed with the postoperative CT examination.Results All patients completed the operation safely.Twenty-eight patients were followed up for an average of 18 months (ranging from 12 to 36 months),and 4 patients who failed to be followed up were not included in the statistic analysis.There was no vessel or nerve injury in the operation.Thirteen out of 28 individuals who suffered from bone cement leakage showed no clinical symptom,and recovered walking ability between 12 and 24h after operation.Other types of vertebral body fractures due to compression occurred in 3 cases between 6 and 12 months after surgery,and the symptoms were relieved after another PKP.The rates of improvement at the last follow-up in VAS and ODI were 71.4％ (20/28) and 67.9％ (19/28),respectively.There was statistical difference in VAS between before surgery and 7d after surgery (t=16.68,P＜0.01).However,no statistical difference was found in the comparison of VAS between the last follow-up and 7d after surgery (t=0.598,P ＞ 0.05).The difference of ODI between before surgery and 7d after surgery (t=36.830,P＜0.01) and that of ODI between the last follow-up and 7d after surgery (t=7.375,P＜0.01) showed statistical significance.There were statistical differences in both Cobb angle and anterior heights of the diseased vertebral body between before surgery and 7d after surgery (t=2.52,2.29,both P＜0.05).Besides,significant differences were also observed in them between the last follow-up and 7d after surgery (t=2.58,2.24,both P＜0.05).However,there was no statistical difference of the posterior height of the diseased vertebral body before surgery,7d after surgery and at the last follow-up (t=0.935,0.795,P＞0.05).Conclusions PKP/PVP may relieve the clinical symptoms safely and effectively of stage Ⅲ Kümmell's disease without nerve injury,and may correct heights of kyphosis and vertebral.Nevertheless,it may lead to loss of correction and high rate of bone cement leakage.Further observation is needed for the long-term effectiveness.