=3) in spinal fluid. The intra and inter-day relative standard deviation of analysis were less than 3.0 % (n = 5). The recovery of lidocaine was between 98.3 % - 102.7 % . Lidocaine assay was carried out in a medical case by using the method established. Conclusion Spinal fluid is suitable for assay lidocain in forensic toxicological analysis and other medical studies by using the HPLC method which is sensitive, rapid and accurate.

BACKGROUND: Study on bone tissue-engineered material is one of the most successful fields in tissue engineering, but the mechanism on synthesis of artificial bone has not been known in many aspects.OBJECTIVE: To explore the mechanism of collagen and calcium phosphate (CP) in artificial bone synthesis.DESIGN: Single sample experiment was designed.SETTING: Material Research Room of Honghe University.MATERIALS: The experiment was performed in Material Research Room of Honghe University from July to August 2003. The materials included collagen (10 g/L acetic acid solution), calcium chloride, sodium dihydrogen phosphate (SDP), sodium hydroxide (NaOH), Tris, hydrochloric acid and deionized water (DI water).METHODS: Liquid nitrogen freezing and freeze-drying were used to prepare collagen-CP complexes A and B and the samples at different times during mineralization. UV spectrophotometer was used to determine the biomineralized dynamic curve of collagen-CP. Based on law of curve, the different times of sample collection were determined in preparation of electronic microscopic samples. According to electronic microscopic pictures and spectral data, mechanism analysis was carried on.MAIN OUTCOME MEASURES: Morphology of collagen-CP complex and law of its structure with time changeRESULTS: ①Under agitation, collagen-CP complex A was sheaf-like or needle-like in structure manufactured with retarded neutralization. ②Under static state, with biomineralization, collagen-CP complex B was in layered structure at initial phase of mineralization, which was similar to the self-assembled structure of pure collagen and the molarratio of C, O, P and Ca was 7.26: 20: 0: 2. At the end of mineralization, the structure was strip-like in high density with a certain grains and very fine rills and the molar ratio of C, O, P and Ca was 11.02: 22.5:1.06: 2.CONCLUSION: At the early phase of biomineralization, collagen iscoordinated initially with calcium ion, calcium-carrier layered collagen template is formed with the self-assembling of collagen, and then phosphates is combined with calcium ion to manufacture calcium phosphate in the formed template. By controlling agitation and acting time, collagen complex material of reticular and spinal structure is obtained.

ObjectiveTo study the effect of anterior decompression and autograft fusion under video-assisted thoracoscopic to treat lower thoracic vertebrae bursting fracture complicated with intervertebral disc injury.MethodsEleven patients who suffered from lower thoracic vertebrae bursting fracture complicated with intervertebral disc injury were treated with anterior decompression and autograft fusion under video-assisted thoracoscopic from December 2005 to May 2008.The involved vertebrae included T12 in 5cases,T11 in 4 cases,T10 in 1 case and Ts in 1 case.According to the AO classification,4 patients were A2.2and 7 were A3.According to the Frankel classification,5 patients were rated as grade A,2 as grade C,2 as grade D and 2 as grade E.ResultsEvery patient underwent successful operation.The operation time was 3.5-8 h(average,5.2±1.6).The blood loss was 600-3800 ml(average,1195±576).One patient got intercostal neuralgia after operation,which disappeared after treating with analgesic drugs for 7 days.All patients were followed up for 36-65 months (average,49.5±5.9).All patients got bony fusion according to the CT scans 24month after operation.There were no neurological function deterioration and other instrument complications happened.At the last follow-up,5 patients were rated as grade A,2 as grade D and 4 as grade E,according to the Frankel classification.ConclusionAnterior decompression and autograft fusion under video-assisted thoracoscopic is an effective method to treat lower thoracic vertebrae bursting fracture complicated with intervertebral disc injury.However,this method needs higher technology and has a long study-curve.

The effects of different concentrations of vascular endothelial cell growth factor (VEGF) on the fibrovascular ingrowth into rabbits hydroxyapatite orbital implant were investigated. Twelve New Zealand white rabbits were divided into 3 groups and received hydroxyapatite orbital implant surgery in their right eyes. Before and after the operation, the implants were treated with 10 ng/ml VEGF, 100 ng/ml VEGF, or normal saline as control group. The animals received technetium bones scan at 2, 4, and 6 weeks postoperatively. The mean radioactivity counts within region of interest (ROI) of the surgery eye (R) and the non-surgery eye (L) in the same animal were tested, and the R/L ratios were calculated. The implants were harvested at 6th weeks and examined histopathologically. The results showed that at second week, there was no significant difference in mean R/L ratios between VEGF group and control group (F=2.83, P=0.111); At 4th week (F=7.728, P=0.011) and 6th week (F=7.831, P=0.011) postoperatively, the mean ratios in VEGF groups were significantly higher than that in control group. At 6th week postoperatively, the fibrovascularization rates in VEGF groups were higher than in control group significantly (F=8.711, P=0.008). It was suggested that VEGF could promote the fibrovascular ingrowth into hydroxyapatite orbital implant, thus might shorten the time required for complete vascularization of the HA orbital implant.
Eye, Artificial ; *Hydroxyapatites ; Neovascularization, Physiologic/*drug effects ; Orbit/blood supply ; *Orbital Implants ; Random Allocation ; Vascular Endothelial Growth Factor A/*pharmacology

Objective To construct eukaryotic expression recombinant plasmid containing human granulysin(GLS) and investigate the effect of GLS expression in macrophage RAW264.7 cells on the bactericidal activity against intracellular Mycobacterium tuberculosis.Methods GLS gene was amplified by nested-polymerase chain reaction(PCR) from human cytotoxicity T lymphocyte(CTL) activated by allogenic antigen,and inserted into pBudCE4.1 vector to construct recombinant plasmid.Subsequently,the plasmid was transfccted into RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The expression of GLS was detected by nested-PCR and immunocytochemistry method.The RAW264.7 cells were lysed after transfected for 96 h,then acidfast stained,cultivated and colony count were done to determine the intraeellular bactericidal activity of GLS.The data were analyzed by t or t' test.Results The pBudCE4.1/GLS eukaryotic expression recombinant plasmid was successfully constructed.The transcriptional and translational expressions of target gene GLS were detected in RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The bacterial load in macrophages of phorbol myristate acetate(PMA)+pBudCE4.1/GLS group,PMA+pBudCEA.1 group and non-activated group were 1.44±1.25,3.16±0.20 and 3.59±0.21,respectively.The differences between groups were all significant (t=2.403,t=2.854,both P＜0.05).Conclusion Eukaryotic expression recombinant plasmid carrying human GLS gene expressed in macrophages has strong bactericidal activity against intracellular mycobacteria,which provide information for the further study on therapeutic vaccine against Mycobacterium tuberculosis.

Objective To investigate the risk factors associated with gastroparesis syndrome after laparoscopic pancreatoduodenectomy which provide reference for clinical prevention.Methods Ninety cases of laparoscopic pancreatoduodenectomy admitted from August 2013 to December 2016 in Traditional Chinese Medicine Hospital of Guangdong Province were studied retrospectively,57 were male(63.3%),the average age was 54.6 years old.Twenty cases were diagnosed postoparative gastroparesis syndrome(22.2%).To screen out the risk factors,31 independent variables were analyzed by univariate analysis and logistic regression.Results Univariate analysis showed that malnutrition,hypoproteinemia,anemia,pylorus-preserving,extensive lymph nodes dissection,anxiety,high blood sugar before operation,delay of enteral nutrition,abdominal infection and postoperative high blood sugar were associated with postoperative gastroparesis(The value of OR were 3.143,3.587,2.852,2.889,3.231,7.071,2.889,5.359,6.000,6.263,P<0.05).Multivariate Logistic regression analysis showed that extensive lymph nodes dissection,anxiety,pylorus-preserving,abdominal infection,delay of enteral nutrition,hypoproteinemia,postoperative high blood sugar were risk factors of postoperative gastroparesis(The value of OR were 17.574,8.931,6.637,6.461,6.446,5.414,5.200;P<0.05).Conclusion Multiple risk factors can lead to gastroparesis after laparoscopic pancreatoduodenectomy,measures should be taken aimed at these risk factors during perioperative period.

Objective To observe the tumorigenicity of High Five insect cell line in Balb/c nude mice,and make sure the safety of the cells when used in vaccine production.Methods Balb/c nude mice were randomly divided into 5 groups:the basic cell bank of High Five group,the highest limited passages of High Five group,HEp-2 cell group as positive control,CEF cell group as negative control,and blank control.Except of the blank control,cell suspension was injected subcutaneously into the nude mice in the different groups,respectively.At 3 and 12 weeks after injection,anatomical observation and histopathologic examination were performed to detect the tumor formation.Results At 3 and 12 weeks after injection,the tumorigenicity study results showed that no tumor developed at the transplantation site in the blank control group,negative group,and High Five groups.Histopathological examinations also showed no abnormality in these groups.Otherwise,squamous cell carcinoma was developed in the positive group at 3 weeks after injection.Conclusions High Five cells of basic cell bank and highest limited passages are not tumorigenic,and can be used in vaccine production safely.

Objective To study the effect of hyperbaric oxygen(HBO) treatment on plasma endotoxin,sCD14 and plasma endotoxin inactivation capacity(EIC) in acute necrotizing pancreatitis(ANP) in rats and its possible mechanism.Methods SD rats were randomly divided into control group,sham operation group,ANP group and ANP+HBO group.Rat ANP models were made by retrograde injection of 3.5% sodium(taurocholate)(2.5mL/kg) into the pancreatic duct.Thirty minues after models had been made,ANP+HBO group was treated by hyperbaric oxygen for 2h.At 3h,6h,and 10h after establishment of rat models,the plasma endotoxin,sCD14,EIC,TXB_2 and 6-K-PGF_(1a) leves were determined in each group.Results At 3h and 6 h after rat models were established,the levels of endotoxin,sCD14,and TXB_2 in ANP+HBO group were significantly lower than those in ANP group(P

Dendritic cell (DC) is professional antigen presenting cell with specific functions. DC can specifically phagocytize antigen, and process and deliver to effector T cell, which play an important role in immune tolerance and immune response. Tolerance DC (tol-DC) is a group of DC with negative immune regulatory function, which can induce immune tolerance of organ transplantation through central and peripheral mechanism. In this article, the phenotype and functional characteristics of DC, mechanism of immune tolerance induced by DC, and the application of tol-DC in organ transplantation of DC were summarized, aiming to provide basic theoretical support for clinical translational medicine research.

Objective To investigate the treatment effects of Kansui root on severe acute pancreatitis (SAP) with abdominal compartment syndrome (ACS).Methods 16 cases of SAP were randomly divided into kansui root treatment group and control group according to random number table.Patients in control group received routine treatment including fasting, anti-shock, antibiotics and nutritional support.And the patients in kansui root group received routine treatment plus kansui root therapy.The clinical and laboratory parameters were determined and compared between the two groups.Results The relieving time of abdominal pain, bowel sound, the recovery time of hyperamylasemia, body temperature and leukocyte count in treatment group was (7.6±2.3)d, (6.1 ±3.1)d, (5.9±3.3)d, (5.2 ±3.2) d, (6.3 ±2.1)d, which were significantly shorter than those in control group [ ( 11.7 ± 2.1 ) d, ( 11.2 ± 2.3d, ( 10.2 ± 2.7) d, (9.2 ± 3.5 ) d, ( 11.1 ±3.3)d, P＜0.01 ) ].At the 3rd, 4th and 5th day, the intra-abdominal pressure in treatment group were also significantly lower than those in control group[ ( 19.8 ±3.1 )cmH2O vs(23.7 ±2.9) cmH2O, ( 12.3 ±2.7) cmH2O vs (21.3±1.5)cmH2O,(8.2±3.1)cmH2O vs (17.3 ±2.3)cmH2O,P＜0.05].Conclusions Severe acute pancreatitis has close relationship with Jiexiong syndrome in traditional chinese medicine.Kansui root is an effective therapy for alleviating high intra-abdominal pressure.