Objective To evaluate the efficacy of plasma radiofrequency ablation at low temperature in disc combined with collagenase injection out of disc in patients with cervical intervertebral disc herniation.Methods Fifty-six patients suffering from cervical intervertebral disc herniation with headache,dizziness,and pain in the neck and in the shoulder were randomly divided into 2 groups ( n =28 each):collagenase injection out of disc group (group C) and plasma radiofrequency ablation at low temperature in disc combined with collagenase injection out of disc group (group R).All operations were carried out under CT guidance.Results At the sixth month of follow up after treatment,the remission rates of headache,dizziness and pain in the neck and in the shoulder were 86％,79％,and 93％ in group C and 96％,93％,and 100％ in group R,.respectively,with significant difference between the two groups ( P ＜ 0.05 ) Conclusion The efficacy of plasma radiofrequency ablation at low temperature in disc combined with collagenase injection out of disc is much better than collagenase single in patients with cervical intervertebral disc herniation.

BACKGROUND: The glial scar formation after spinal cord injury in mammals is the physical and chemical barriers for neural regeneration, and relieving or delaying glial scar formation can provide benefit conditions for the regeneration of injured spinal cord.OBJECTIVE: To design and screen short hairpin RNA (shRNA) interfere molecular targeting the gene coding region of glial fibrillary acidic protein (GFAP) in rat, and reconstruct the eukaryotic vector of shRNA.DESIGN: An observational animal experiment.SETTING: Department of Spine Surgery, Shenzhen Hospital affiliated to Southern Medical University.MATERIALS: Twenty-five Wistar rats of clean degree, either male or female, weighing 20-25 g, were used. DMEM/F12,lipofectamine2000, Trizol RNA separating kits); fetal bovine serum (Hyclone); BamH Ⅰ, HindⅢ, Pstl, Sail and T4 ligases;Plasmid mini preparation kit and DNA gel extraction kit.METHODS: The experiments were carried out in the Shenzhen Hospital affiliated to Southern Medical University from October 2005 to June 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of GFAP target sequence with 9 bp spacer were designed and synthesized, then they were inserted directionally into plasmid Psilencer 2.1 respectively to generate small interfering RNA (siRNA) eukaryotic expression vector. ShRNA molecules were transfected by liposome via ex vivo expression repressive model of GFAP of rat spinal astrocytes. The effects of expressive suppression were detected with real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, and then the optimal shRNA eukaryotic vector of repressive expression of GFAP was screened.MAIN OUTCOME MEASURES: ① Interfering sequence specific shRNA template synthesis; ② Constructing specific recombinant plasmid eukaryotic expression vector. ③ Culturing rat spinal astrocytes in vitro; ④ Effects of expressive suppression on GFAP in primary astrocytes after RNA interference detected with real-time fluorescence quantitative RT-PCR and Western blot.RESULTS: Sequence analysis showed that GFAP-shRNA recombinant plasmid eukaryotic expression vector was successfully constructed, and optimal GFAP-shRNA eukaryotic vector was screened using real-time fluorescence quantitative RT-PCR and Western blot. The GFAP expressions were reduced by 81%, 63% and 56% at the levels of mRNA and protein respectively.CONCLUSION: GFAP-shRNA eukaryotic expression vectors were successfully constructed and screened. The gene expression GFAP of primitive rat astrocyte can be suppressed significantly by the GFAP-shRNA eukaryotic expression recombinant optimized via ex vivo cellular expression suppression model, which should pave the way for the following multiple targets of RNAi genetic manipulation in the treatment of suppression of glial scar formation after spinal cord injury.

Objective To compare the severity of kidney injury following liver transplantation performed under sevoflurane versus propofol combined anesthesia in patients.Methods Fifty ASA Ⅱ or Ⅲ patients of both sexes,aged 40-64 yr,weighing 47-85 kg,scheduled for elective orthotopic liver transplantation,were randomly divided into 2 groups (n =25 each) ∶ propofol combined anesthesia group (group P) and sevoflurane combined anesthesia group (group Se).During maintenance of anesthesia,propofol was given by target-controlled infusion and the target plasma concentration was 2-4 μg/ml in group P,and sevoflurane was continuously inhaled and the endtidal concentration was maintained at 1.0％-3.0％ in group Se.Venous blood samples and urine specimens were taken immediately before skin incision (T0),at 30 min after occlusion of the inferior vena cava (T1),at 30 min of neohepatic phase (T2),at the end of surgery (T3),and at 1,3 and 5 days after operation (T4-6) for determination of serum concentrations of creatinine (Cr),blood urea nitrogen (BUN),β2-microglobulin (β2-MG) and urinary β2-MG concentrations.Results Compared with group P,the serum concentrations of Cr,BUN and β2-MG and urinary β2-MG concentrations were significantly lower at T3,4 in group Se (P ＜ 0.05 or 0.01).Conclusion The severity of kidney injury is reduced in patients undergoing liver transplantation performed under sevoflurane combined anesthesia compared with that under propofol combined anesthesia.

Objective To investigate the effect of creatine phosphate on perioperative myocardial injury caused by living donor liver transplantation(LDLT)in adult patients.Methods Forty ASA Ⅱ -Ⅳ patients(liver function Child-Pugh grade B or C)aged 45-62 yr weighing 47-91 kg undergoing LDLT were randomly divided into 2 groups(n = 20 each): control group(group C)and creatine phosphate group(group CP).In group CP,creatine phosphate 30 mg/kg was injected intravenously at skin incision followed by creatine phosphate infusion at 4 mg· kg- 1 · h- 1 until the end of surgery.In group C,equal volume of normal saline was infused instead of creatine phosphate.HR,MAP,CVP,PCWP,CO and SvO2 were recorded immediately before skin incision,at 5 and 30 min of anhepatic phase,at 5 and 30 min of neohepatic phase and at the end of operation.Blood samples were taken from central vein immediately before skin incision(baseline,T0),at 30 min of anhepatic phase(T1),at 30min of neohepatic phase(T2),at the end of operation(T3)and at 4 and 24 h after operation(T4,5)for determination of serum cardiac troponin I(cTnI)and creatine kinase MB(CK-MB)concentrations and lactate dehydrogenase(LDH)activity.Postoperative adverse events were recorded.Results The serum cTnI and CK-MB concentrations and LDH activity were significantly increased at T2-5 as compared with the baseline value at T0 in both groups(P ＜0.05 or 0.01).MAP and CO were significantly higher from 5 min of neohepatic phase to the end of operation,the serum cTnI and CK-MB concentrations and LDH activity were significantly lower at T2-5,and the incidence of ventricular arrhythmia was significantly lower in group CP than in group C(P ＜ 0.05 or 0.01).Conclusion Creatine phosphate can attenuate perioperative myocardial injury caused by LDLT in adult patients.

Objective To investigate the effect of interleukin-1 (IL-1) on contractile function of rat thoracic aorta.Methods Forty male Sprague-Dawley rats,weighing 250-300 g,were sacrificed to obtain the thoracic aortic rings.The experiment was performed in 2 parts.Part Ⅰ The thoracic aortic rings were divided into 2 segments and randomly divided into 2 groups (n =20 each):control group and IL-1 group.In IL-1 group,the thoracic aortic rings were incubated with Kreb solution containing 20 ng/ml IL-1 for 2 h,and contracted with cumulative concentrations of phenylephrine,ranging from 10-9 to 10-5mol/L.In control group,the thoracic aortic rings were incubated with Kreb solution for 2 h,and contracted with cumulative concentrations of phenylephrine,ranging from 10 9 to 10-5mol/L.Part Ⅱ The thoracic aortic rings were divided into 3 segments and randomly divided into 3 groups (n=20 each):IL-1 group,IL-1+ L-NAME (the NOS inhibitor) group and IL-1 +cyclooxygenase inhibitor indomethacin group (IL-1 +Ⅰgroup).The thoracic aortic rings were incubated with Kreb solution containing 20 ng/ml IL-1 for 1.5 h in the three groups.In addition,in IL-1 +L-NAME and IL-1 +Ⅰ groups,the thoracic aortic rings were incubated for 30 min with Kreb solution containing 100 μmol/L L-NAME and 2.5 mmol/L indomethacin,respectively.Contraction of the thoracic aorta was then induced with cumulative concentrations of phenylephrine,ranging from 10 9 to 10-5 mol/L.In group IL-1,the thoracic aortic rings were incubated with Kreb solution.The maximum contractile tension of the thoracic aortic rings was recorded at each concentration of phenylephrine,and the percentage of the maximum contractile tension at the concentration of 10-6 mol/L in group C was obtained.Results Part Ⅰ The percentage of contractile tension at phenylephrine 10-s,10-7,l0 6 and 10-5mol/L was significantly decreased in IL-1 group as compared with C group.Part Ⅱ The percentage of contractile tension at phenylephrine 10-7,10-6 and 10-5mol/L was significantly increased in IL-1+L-NAME and IL-1+I groups as compared with IL-1 group.Conclusion IL-1 can inhibit the contraction of rat thoracic aorta,and promoted production of NO and prostacyclin may be involved in the mechanism.

Objective To study the effect of the recombinant vaccinia virus expressing human interleukin 2 on MKN45 gastric cancer cells in vitro. Methods Recombinant vaccinia virus expressing hIL 2 (VMJ601hIL 2) was constructed by homologous recombination using molecular virology, and VMJ601hIL 2 was detected by DNA hybridization technique and the recombinant gene product was analyzed by SDS PAGE. In addition, MKN45 gastric cancer cells was infected by VMJ601hIL 2 and the effect of VMJ601hIL 2 on the gastric cancer cells was evaluated in vitro. Results hIL 2 protein with biological activity can be secreted by MKN45 gastric cancer cells after heavily infected by VMJ601hIL 2. Conclusions It is one of the crucial steps that VMJ601hIL 2 has been constructed and identified since it forms essential prerequisite for further in vivo gene therapy of gastric cancer.

Objective To investigate the blood conservation techniques during orthotopic liver transplantation(OLT). Methods Thirty-eight patients undergoing OLT without veno-venous bypass under general anesthesia were studied. During operation, the blood conservation measures such as controlled blood loss and blood coagulation, etc. were implemented. The input and output of liquid, blood component transfusion in preanhepatic, anhepatic and reperfusion phases was analyzed. The blood samples from central vein were collected, plasma creatinine, hemoglobin and albumin were determined after anesthesia, 1 h after preanhepatic phase, 20 min after anhepatic phase and 1 h after reperfusion phase, and the central venous pressure and Sonoclot data were measured.Results The total volume of whole blood transfusion was about ( 816? 86.3) ml and that of red blood cells ( 962? 55.3) ml during operation. Two patients had no blood transfusion. The normal urinary output was maintained during preanhepatic, anhepatic and reperfusion phases. The central venous pressure at three phases was significantly decreased compared to that after anesthesia (P 0.05). The plasma creatinine was significantly increased at reperfusion phase (P

Objective To make a passive personal sampler for the determination of the styrene in the air. Methods To make a passive personal sampler for the determination of the styrene in the air. Results Under the condition of wind velocity of 100-600 cm/s, relative humidity of 40%-80% and temperature of 10-40 ℃, the sampling flux of the passive sampler was 61.8 ml/min, the capacity was 5.95 mg, the shortest sampling time was 30 min, the self life was at least 30 days, the storage stability was at least 14 days, the RSDs were 8.07%-9.37%. Conclusion This new kind of passive personal sampler can be used as a new device for the determination of the styrene in the air.

Objective To explore the effect of drug injection in muscle motor points on children with spastic cerebral palsy. Methods 230 children with spastic cerebral palsy were divided into control group and observation group, with 115 cases in each group. The control group received comprehensive rehabilitation including functional training, physical therapy and medicine, while the observation group received injection of vitamin B1 and vitamin B12 in muscle motor points in addition. They were assessed with Gross Motor Function Measure (GMFM), modified Ashworth Scale (MAS), mental development index (MDI) and psychomoter developmental index (PDI) of CDCC Infant Me Ntal Development Scale. Results The gross motor function improved, MAS score decreased, the scores of MDI and PDI increased after treatment (P<0.01) in both groups, which were better in the observation group than in the control group (P<0.05). Conclusion Drug injection in muscle motor points can improve the gross motor function, relieve spasm, and improve intelligence development of children with spastic cerebral palsy.

ObjectiveTo investigate the effects of functional point injection on upper limbs disorder in cerebral palsy infants. Methods120 cases were divided into 2 groups and each with 60 cases. The treatment group was given rehabilitation and functional point injection (injecting VitB1>, VitB12 in the acu-points selected at the disorder muscles) and the control group was given rehabilitation alone. All the cases were assessed with Peabody Developmental Motor Scale Fine Motor (PDMS-FM) and Modified Ashworth Scale (MAS). ResultsThe scores of both PDMS-FM and MAS improved in the treatment group compared with the control group (P<0.05). ConclusionFunctional point injection can improve the recovery of upper limbs disorder in cerebral palsy infants.