Using eccentric wheel and spring, an instrument was developed in this study. Relative information of the instrument were clearly explained, including its structure and running principle. The instrument can be widely used in fatigue measurement of fixed prostheses and capability of resisting break, and provide the experimental gist of the using of prostheses.

Objective To investigate effects of parecoxib sodium preemptive analgesia on postoperative analgesia and cellu‐lar immune function in patients with uterine fibroids.Methods Totally ,116 cases of patients receiving uterine fibroids surgery were selected from March 2011 to April 2014 in our hospital.All cases were randomly divided into advanced group(n=58)and control group(n=58). The cases in the advanced group were intravenously injected with parecoxib sodium(40 mg)30 min before anesthesia ,while the control group received 5 mL saline intravenously.Postoperative pain conditions of the two groups were e‐valuated by using visual analog scale(VAS method). The PCIA pump first trigger time ,effective compression number and the total amount of fentanyl were recorded. Venous blood was collected before operation ,6 h(T1 ) ,24 h(T2 ) ,48 h(T3 )and 72 h(T4 ) after operation ,respectively. T lymphocyte subsets of CD3+ ,CD4+ and CD8+ and NK cells were detected by using FACS Cali‐bur flow cytometer.The ratio of CD4+ /CD8+ was calculated.Results After 4 ,8 ,12 ,24 and 48 h ,the pain scores of patients were significantly lower in the advanced group than in the control group(all P<0.05). The PCIA pump first trigger time was longer ,effective compression number and the total amount of fentanyl of patients were less in the advanced group than in the control group ,the differences were statistically significant(all P<0.05). At T1 ,T2 and T3 ,CD3+ and CD4+ were significantly higher in the advanced group than in the control group(P<0.05).At T1 and T2 ,CD4+ /CD8+ was significantly higher in the advanced group than in the control group(both P<0.05).At T2 and T3 ,the NK was significantly higher in the advanced group than in the control group(both P<0.05).Conclusion Parecoxib sodium preemptive analgesia used in patients with uterine fi‐broids can reduce postoperative pain ,reduce the amount of analgesics ,reduce the immune suppression ,and improve immune function in patients after surgery.

objective: To generate 3 D model of teeth for finite element method(FEM) analysis by computer. Methods: On the basis of 3 D model created by 3D Max, the 3 D FEM model of teeth was generated by using of CAE software. Boundary condition was set in the part of teeth root with elastic constraint. The elastic modulus was chosen as the modulus of periodontium. Results: Generated right inciser model comprised of 25 386 units and 5 111 nodal points. Conclusion: 3 D FEM model of tooth can be generated by CAE software.

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.

Bioactive peptides in the marine organisms such as sponges,ascidian,fishes,shellfish,etc. and their bioactivities of antineoplastic, antimicrobial, antihypertension and antioxidation were reviewed in this paper.

Objective To evaluate the clinical value of SCTPA and pulmonary V/Q scan in the diagnosis of pulmonary thromboembolism(PTE).Methods Ninety-two patients suspected of having PTE received SCTPA,pulmonary V/Q scan,and other related examinations.Results Thirty-five patients were diagnosed as having PTE in 92 patients investigated,30 cases were revealed by SCTPA,and 20 cases revealed by pulmonary V/Q scan.The area under ROC curve of SCTPA by V/Q scan and combination examination was 0.922,0.824,and 0.933,respectively(P

BACKGROUND: Agiotensin Ⅰ -converting enzyme (ACE) inhibitory peptides which are separated from red tilapia skin collagen should be researched further.OBJECTIVE: To obtain a peptide of high ACE inhibitory activity through enzymic hydrolysates of red tilapia skin collagen.DESIGN: Enzymic hydrolysates were done with orthogonal experimental method; decompression peptide was separated with ultrafiltration, gel chromatography and hypertensive liquid chromatography.SETTING: Laboratory of Aquatic Products, Chinese Marine University.MATERIALS: The red tilapia weighing (500+50)g was donated by Branch Factory of Jimo Redian Factory. ACE was isolated from hog lung.METHODS: The experiment was carried out in the Laboratory of Aquatic Products, Chinese Marine University from May Bergman.①The collagen from red tilapia skin was extracted using the method described by Grossman and Bergman.The collagen extraction was hydrolyzed with compound enzymes in the order of bromelain and Alcalase 2.4 L.The red tilapia collagen hydrolysates (RTCH) were subsequently boiled to inactivate the enzyme. The resultant RTCH was fractionated through three different UF membrane bioreactor system having a range of molecular weight cut-offs (MWCO) of Mr 6 000, 4 000 and 1 000. Three portions were obtained: RTCH-I (M, 6000-4000), RTCH- Ⅱ (Mr 4000-1000)and RTCH-Ⅲ(Mr＜1000).②The ACE inhibitory 50%of ACE activity was defined as the IC50 value.③RTCH-Ⅲ(1.5 Ml) with the highest activity among RTCHs was loaded onto a Sephadex G-25 column (1.6×100 cm), and the absorbance of theeluent was monitored at 220 nm. Four samples of primary hydrolysates, RTCH- Ⅰ, RTCH- Ⅱ and RTCHⅢ were separated to collect active fractions which were pooled and lyophilized, immediately. The lyophilized fraction was separated with HPLC ODS-C18 analysis column to obtain component of high activity. Meanwhile, the same method was used to measure ACE inhibitory rate.④Each sample was hydrolyzed with 6 mol/L hydrochloric acid containing 1g/L thioglycolic acid at 110 ℃ for 24 hours in vacuum. The amino acid compositions of the resultant hydrolysates were determined on an amino acid auto analyzer, and molecular weight was measured with HPLC technique.MAIN OUTCOME MEASURES：①ACE inhibitory activity of fractionated RTCHS;②Purification of ACE inhibitory peptide;③Amino acid analysis of ACE inhibitory peptides.RESULTS:①ACE inhibitory activityof fractionated RTCHs:IC50values of RTCH-Ⅰ,RTCH-Ⅱand RTCH-Ⅲ were 3.30,2.23 and 0.31 g/L,and inhibitory of RTCH-Ⅲ was the highest.②Purification of ACE inhibitory peptide: The IC50 value of the four peak were 3.5, 2.12, 1.56 and 0.65 g/L, respectively. Results in Figures 2, 3, 4 and 5 showed that the high active fractions were: 6K4, 4K2, 1K2 and ACF2, and the inhibitory ratio were 11.1%, 89.9%, 65.0% and 49.7%, respectively.Among of these fractions,the 4K2 exhibited the highest inhibitory rate.③Amino acid analysis of ACE inhibitory peptides: Separated peptide products had more aromatic and hydrophobic amino acids.CONCLUSTON: Enzymic hydrolysates of red tilapia skin collagen have high ACE inhibitory peptide which is separated with ultrafiltration, gel chromatography and hypertensive liquid chromatography.

BACKGROUND: It is demonstrated that bone marrow stem cells (BMSCs) can generate neurosphere structures, which is similar to cloning sphere of neuron-specific enolase (NSE), in a specially induced system in vitro; therefore, BMSCs draw more and more attention as seed cells to repair central nerve injury.OBJECTIVE: To investigate the differentiation from BMSCs into neuron-like calls induced by fetal spinal cord tissue.DESIGN: Observational study.SETTING: Department of Spine Surgery, Second People's Hospital of Shenzhen.MATERIALS: SD rats (16 pregnant days old and 2 months old) were provided by Animal Center, Tongji Medical College,Huazhong University of Science and Technology. Single antibody of NSE, multi-antibody of glial fibrillary acidic protein (GFAP) and single antibody of neurofilament (NF200) were provided by Wuhan Boster Company.METHODS: The experiment was carried out in Immune Opening Laboratory and Central Laboratory, Basic Medical Department, Tongji Medical College, Huazhong University of Science and Technology in September 2006. Bones of lower extremities of rats were collected to centrifuge BMSCs. Fetal spinal cord tissue homogenate was extracted from 16 pregnant days old rats to make inducing solution and induce differentiation of BMSCs. Otherwise, embryo muscle tissue was used to make muscle tissue homogenate as the same way so as to regard as the controls.MAIN OUTCOME MEASURES: BMSCs underwent morphological observation after induction; in addition, anti-NSE, NF200 and anti-GFAP were used to label neurons and astrocytes, respectively. Ten non-overlapping sights were randomly selected from positive reactive-induced cells after immunohistochemical staining under optic microscope to calculate ratio of positive cells of NSE and NF200 counting for total numbers of cells.RESULTS: ① Morphological changes of BMSCs after induction: During early induction, optic microscope indicated that soma of partial cells rebounded; whose apophysis was long and thin; apophysis of differentiated cells grew gradually and intercrossed each other. It was similar to nerve cells and some branches were similar to dendrite branches. However,morphological changes of cells in the control group were not obvious. ② Expression of relevant antigens differentiated from BMSCs into neuron-like cells at one week after induction: Most cells in spinal cord homogenate group expressed as positive NSE and NF200; a few of cells expressed as GFAP. While, positive staining of nerve cell antibody was not observed in the control group; meanwhile, positive reaction of nerve cell antigen was not observed in the control group,too. Immunohistochemistry examination demonstrated that positive rates of NSE and NF200 expressions were (68±1.7)%and (76.2±2.9)%, respectively.CONCLUSION: Fetal spinal cord tissue homogenate can induce differentiation from BMSCs into neuron-like cells.

Objective To investigate the effect of small direct‐current electrical stimulation on migration and invasion related MMPs/TIMPs expression of trophoblast cells .Methods The trophoblast cells were exposed to the direct current electrical field at 150 mV/mm for 5 and 10 hours .Cell images were recorded with continuous photographing and analyzed by image analyzer .The ex‐pression levels of MMP2 ,MMP9 ,TIMP1 and TIMP2 were measured using quantitative RT‐PCR and Western blot .Results In non‐electrical field culture trophoblast cells migrated slowly with random directions .Trophoblast cells cultured in media containing 10% calf serum with the application of 150 mV/mm direct current electrical stimulation ,showed marked cathodal migration (P<0 .01) ,the cell body stretched ,perpendicular to the direction of the electric field .Compared with the non‐electrical field stimulation controls ,trophoblasts under the electrical field stimulation had the increased MMP2 mRNA and protein expression (P< 0 .05) , while MMP9 ,TIMP1 and TIMP2 had no obvious changes of mRNA or protein expressions .Conclusion Physiological direct‐cur‐rent electrical fields might induce directed migration and perpendicular orientation of trophoblast cells .The enhanced MMP2 expres‐sion may play an important role in the migration and invasive activity of trophoblast cells in small electrical field .