=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.

Objective To detect micronuclei (MN) induced by nickel chloride (NiCl2) using a flow cytometry-based method, so as to study the mutagenecity of this chemical. Methods The micronucleated reticulocytes (MN-RET) in peripheral blood of NIH mice exposed to 0.75 mg/kg colchicines (COL), 40.0 mg/kg of cyclophosphamide (CP) and 5.0, 10.0 and 20.0 mg/kg of NiCl2 were detected by a flow cytometry-based method, in which anti-CD71-FITC and PI were used to stain cells and malaria-infected erythrocytes were employed as biological standards of micronuclei to optimize cytometry. Results Although COL or CP induced significant increase in the frequency of MN-RET in peripheral blood, MN formation in peripheral reticulocytes was not significantly affected by exposing to different concentration of NiCl2. Conclusion The results of flow cytometric analysis in the present paper do not support that NiCl2 exposure may induce significant MN-RET increase in peripheral blood in NIH mice.

Both molecular weight 6000 and 1000 polyethylene glycol (PEG) have been used to induce thc fusion of hepatoma cell by two methods of centrifugation and stillstand. The results showed that 50% PEG of molecular weight 1000 and centrifugation increase the percentage of fused cells. The procedure of cell fusion has been examined. The cytoplasm has been fused when the cells start fusing in one hour and a half, and the neucleus start fusing in two horus and a half.

Objective: To study the protective effect of compound extract of soybean, wolfberry and hawthorn on chemical liver injury in mice respectively. Methods:The mice were randomly divided into 5 groups:control(A), liver injury control(B)and 0.06(C)?0.20(D)?0.60(E) g/(kg?bw) compound extract group. The liver injury models were induced by CCl 4 and ethanol in mice respectively. After 4 weeks, for the former, the serum ALT and AST content were measured and the liver pathological change was observed. For the latter, the liver MDA, GSH, and TG content were measured and fat droplets staining in the ice pathological slice were observed . Results: 1.CCl 4 liver injury models:the content of serum ALT of group D and the AST of group D and E were decreased. Compared with normal group, the hepatic pathological changes of the compound extract group was mainly fatty and ballon degeneration, and that of the CCl 4 group was mainly hepatic cytoclasis. 2.Ethanol liver injury models: the content of liver MDA of group C, E and the TG of group C, D, E were all decreased, and the fatty degeneration scores of group C, D were lower, both compared with ethanol group. Conclusion: Compound extract of soybean, wolfberry and hawthorn has the protective effect on liver injuries induced by both CCl 4 and ethanol in mice.

Objective To investigate the diagnostic value of microscopic observation drug susceptibility assay (MODS) in extrapulmonary tuberculosis. Methods MODS technology was constructed by using 24-well cell culture plate and liquid culture.Ziehl-Neelsen smear,Lowenstein-Jensen culture and MODS were used to detect Mycobacterium tuberculosis in 74 pleural fluid samples collected from patients with tuberculous pleurisy,63 cerebrospinal fluid samples collected from patients with tuberculous meningitis and 18 samples collected from non-tuberculosis suspects.The immunochromatography was used to distinguish Mycobacterium tuberculosis from nontuberculosis mycobacteria. The results of Ziehl-Neelsen smear, Lowenstein-Jensen culture and MODS were compared by x2 test.Results The positive rates of MODS,Lowenstein-Jensen culture and Ziehl-Neelsen smear were 58.1 ％ (43/74),18.9 ％ (14/74 ) and 6.8％ (5/74),respectively in tuberculous pleurisy patients; 54.0％(34/63),20.6％ (13/63) and 4.8％ (3/63),respectively in tuberculous meningitis patients.The positive rate of MODS technology was significantly higher than that of Lowenstein Jensen culture in tuberculous pleurisy patients (x2 =24.00,P＜0.01) and tuberculous meningitis patients (x2 =14.97,P ＜ 0.01). Each Mycobacterium obtained from MODS and Lowenstein-Jensen culture was identified as Mycobacterium tuberculosis by immunochromatography.All of the 18 pleural fluid and cerebrospinal fluid samples which collected from non-tuberculosis suspects were all negative detected by Ziehl-Neelsen smear,Lowenstein-Jensen culture and MODS.The median time to culture positive of MODS was 9 days in cerebrospinal fluid and 14 days in pleural fluid samples,which were both significantly shorter than that of Lowenstein-Jensen culture (31 days in both cerebrospinal fluid and pleural fluid samples). Conclusion Compared to conventional microbiological diagnosis methods,MODS is a rapid detection method of Mycobacterium tuberculosis with a higher positive detection rate,which is suitable for rapid diagnosis of extrapulmonary tuberculosis.

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.

Objective To investigate the safety and feasibility of fast track surgery (FTS)in patients with vaginal hysterectomy for non -prolapsed uterus.Methods 1 1 0 cases of vaginal hysterectomy for non -prolapsed ute-rus with benign uterine disease were randomly divided into two groups:fast track group (n =55)and conventional group (n =55).The operative time,intraoperative blood loss,postoperative nausea and vomiting,anal exhaust time, postoperative hospital stay,medical cost and perioperative complications were compared between the two groups. Results In FTS group,the incidence rate of postoperative nausea and vomiting,the postoperative anal exhaust time, hospital stay,cost of hospitalization were 1 6.4%,(1 3.73 ±2.41 )h and (4.38 ±1 .08 )d,(7 541 .00 ± 253.1 7)yuan,respectively,which in the control group were 36.4%,(1 8.56 ±1 .54 )h,(4.89 ±1 .26 )d, (8 1 55.1 5 ±495.89)yuan,the differences were statistically significant between the two groups (χ2 =5.67,t =-1 2.53,-2.28,-8.1 7,all P <0.05).There was no significant difference in the operative complications between the two groups (P >0.05).Conclusion FTS has good security.It can short the hospitalization time,reduce medical costs and improve the quality of life in patients who underwent vaginal hysterectomy for non -prolapsed uterus with benign uterine disease.

AIM: To investigate the effects of TGF-?_1 and signal protein Smad3 on rat cardiac myocyte hypertrophy.METHODS: The total protein was analyzed by flow cytometry and the ANF mRNA expression was measured by RT-PCR to judge the hypertrophy of cultured neonatal cardiac myocytes.Smad3 mRNA expression in cardiac myocytes was measured by RT-PCR,and the protein expression of Smad3 was analyzed by Western blotting.RESULTS: TGF-?_1 significantly increased the total protein in cardiac myocytes and promoted ANF mRNA expression,compared with control group.In cultured neonatal myocytes,AS-ODN of Smad3 inhibited myocyte hypertrophy induced by TGF-?_1.Smad3 mRNA and protein expression increased at 15 min after incubated with TGF-?_1,reached the peak at 1 h,and declined at 4 h.CONCLUSION: TGF-?_1 and signal protein Smad3 may participate in the progress of rat cardiac myocyte hypertrophy.

Objective:To explore changes of peripheral blood levels of C reactive protein (CRP)and matrix metallo-proteinase-9 (MMP-9)in patients with coronary heart diseases (CHD)and percutaneous coronary intervention (PCI),and analyze their relationship with patients'prognosis.Methods:A total of 278 CHD patients undergoing PCI in our hospital from Jul 2009 to Apr 2011 were regarded as PCI group;another 234 CHD patients not receiving PCI were enrolled as CHD control group.According to results of coronary angiography,PCI group was further di-vided into single-vessel (n=143),double-vessel (n=92)and triple-vessel disease group (n=43).Changes of CRP and MMP-9 levels were compared between two groups in different time,and the relationship among these two in-flammatory factors and coronary disease,its prognosis was analyzed.Results:Compared with on admission,there were significant rise in levels of CRP [(2.43±0.62)mg/L vs.(2.87±0.73)mg/L,(2.98±0.87)mg/L]and MMP-9 [(12.63±2.68)ng/ml vs.(14.62±3.49)ng/ml,(19.62±4.63)ng/ml]in PCI group on 24h and 48h after PCI,P <0.05~<0.01;there were no significant difference in CRP and MMP-9 levels between on admission and 14d after PCI,P >0.05;The more severe coronary lesion was,the higher CRP and MMP-9 levels were,CRP and MMP-9 levels of triple-vessel group [(2.51 ±0.64)mg/L,(14.67±2.97)ng/ml]were significantly higher than those of single-vessel group [(1.83±0.51)mg/L,(9.68±1.42)ng/ml]and double-vessel group [(2.17±0.59) mg/L,(11.62±2.19)ng/ml],P <0.05~<0.01;incidence rates of cardiovascular events in patients with CRP≥3 mg/L and MMP-9≥15 ng/ml (33.3%,29.1%)were significantly higher than those of patients with CRP <3 mg/L and MMP-9<15 ng/ml (16.1%,18.2%)respectively,P <0.05 both.Conclusion:Serum levels of CRP and MMP-9 significantly rise in CHD patients on 48h after PCI,and their increasing level is related with extent of coronary ar-tery lesion and prognosis.

Objective To construct a recombinant bacterial vaccine which can express specific 10-23 deoxyribozyme(DZ) in macrophage, identify the intracellular production of specific 10-23DZ and detect the activity of this recombinant bacterial vaccine on inhibiting the expression of TACO gene in macrophage.Methods The pSDE02 was obtained by inserting the replicon of Mycobacterium into pSDE01, a plasmid which can express 10-23DZ in eukaryotic cells. The expression sequence of DZ1, a 10-23DZ targeting the TACO mRNA of macrophage designed in our previous study was synthesized and inserted into pSDE02. The resulted plasmid was named pDZM01. pDZM01 was then transferred into Mycobacterium smegmatis by electroperation. The recombinant M. smegmatis, named rMs-DZ1 was screened on low-salt LB medium containing Zeocin and identified by Colony PCR. The targeted delivery property of recombinant M. smegmatis was observed by Ziehl-Heelson stain and GFP expression observation via fluorescence microscope. rMs-DZ1 was used to infect RAW264.7 cells and the expression of DZ1 in macrophage was identified by dot-blot assay. At 24 h and 48 h after infection, total RNA and proteins were extracted and the TACO mRNA and protein expression level was assayed by RT-PCR and western-blot respectively. Results Restrictive analysis and sequencing data showed that the Mycobacterium-eukaryotic cell shuttle plasmid pSDE02 and pDZM01 was successfully constructed. rMs-DZ1 was confirmed by colony PCR. When engulfed by macrophage, rMs-DZ1 would express DZ1 in RAW264.7 cells and inhibit the expression of taco gene. When compared to uninfected macrophage, rMs-DZ1 significantly reduced the taco mRNA by 67.90% and 57.14% and down-regulated the expression of TACO protein by 53.85% and 68.92% at 24 h and 48 h respectively. Conclusion A recombinant M. smegmatis vaccine was successfully constructed which could generate specific 10-23DZ in macrophage and inhibit the expression of target gene of interest. To our knowledge, this is the first bacterial vector which can express intracellularly 10-23DRz in targeted manner. This study may further prompt the feasibility of using 10-23 DNAzyme to achieve effective and targeted gene silence.