Objective To construct and identify the specific interference vector for second mitochondria -derived activator of caspase (SMAC)gene in mice and perform lentiviral packaging.Methods According to the SMAC gene sequences in mice,three small interfering RNAs (siRNAs)(siRNA1,siRNA2,and siRNA3)and one negative control sequence (siRNAn)were designed and synthesized,and mouse hepatocytes were transfected with the above siRNAs using Lipofectamine 2000.The inhibitory effects of these siRNAs on SMAC mR-NA expression were evaluated by real-time PCR,and the optimal siRNA was screened out accordingly.Oligo DNA was synthesized based on the optimal siRNA and then connected to GV1 15 vector to obtain recombinant interference plasmid.The candidate clones were identified by PCR and sequenced.The recombinant interference plasmid,as well as pHelper 1.0 and pHelper 2.0,was used to transfect 293T cells for lentivirus packaging.Then,cell supernatants were collected and concentrated,and the lentiviral titer was determined by gradient dilution. Comparison between groups was made by analysis of variance,and multiple comparisons were made by SNK-q test.Results The three siRNAs had different inhibitory effects on SMAC mRNA expression in hepatocytes;siRNA1 showed the strongest inhibitory effect,with an inhibition efficiency of 70.3%.Based on siRNA1 ,the oligo DNA was successfully synthesized and correctly connected to the lentiviral vec-tor,as proved by sequencing.The lentiviral titer was 6 ×108 TU/ml,as determined by gradient dilution.Conclusion The recombinant lentivirus that inhibits SMAC expression in mice has been constructed successfully,laying the foundation for further studies on the role of SMAC gene in liver failure.

BACKGROUND: Advanced oxidation protein products (AOPPs) are a crucial pathogenic link to such long-term uremic complications in hemodialysis patients as immune system dysregulation, accelerated atherosclerosis, dialysis-related amyloldosis and so on. However, basic studies on AOPPs are relatively few, and one of the main reasons is the fact that it is difficult to obtain AOPPs with high pudty and biological activity.OBJECTIVE: To prepare, pudfy and indentify AOPPs, with the hope of searching for a method of preparing AOPPs with high purity and biological activity.DESIGN, TIME AND SETTING: A single sample observation was completed in the Clinical Biochemistry Section of Ecsomatics Department, Third Military Medical University of Chinese PLA from September to November in 2008. MATERIALS: Human serum albumin (HSA) was provided by Chengdu Rongsheng Company Ltd. Hitrap 26/60 sephacryl S-300 was purchased from GE Healthcare.METHODS: Hypochloric acid was used in the oxidation of purified HSA to prepare in vitro the AOPPs-modified HSA (AOPPs-HSA), which was then isolated by Hitrap 26/60 sephacryl S-300. Relative molecular mass was determined by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE), native polyacrylamide gel electrophoresis (PAGE) and molecular weight standards. Structural features and biological activities were identified in the experiment of tumor necrosis factor α (TNF-α) release from monocytes.MAIN OUTCOME MEASURES: ①The purification and gel electrophoresis results of HSA. ②The purification and gel electrophoresis results of AOPPs. ③The dose-effect relationship between AOPPs-HSA and TNF-α release from monocytes. RESULTS: The relative molecular mass of AOPPs-HSA was 670 000 according to SDS-PAGE, native PAGE and molecular weight standards. Moreover, AOPPs-HSA could encourage the release of TNF-α from monocytes. The time effects showed that TNF-α release volume significantly increased after 6 hours of stimulation by AOPPs-HSA (1 g/L) and reached a peak at hour 12. CONCLUSION: Highly purified and bioactive AOPPs can be successfully prepared by the above-mentioned method, which builds a basis for further study on AOPPs.

Objective To determine the effect of treadmill exercise on expression of matrix metalloproteinase-2 (MMP-2) in rats follow-ing traumatic brain injury (TBI). Methods Fifty-five male adult Sprague-Dawley rats were randomly assigned to sham (n=15), control (n=20) and exercise (n=20) groups, the later two groups subjected to unilateral cortical contusion injury (CCI). All the rats were assessed with foot-fault test 6, 12, 18 and 24 days after CCI. Evans blue perfusion was used to evaluate the integrity of the blood-brain-barrier (BBB) 48 hours after CCI. Protein expression of MMP-2 was determined with Western blotting one week after CCI. Results The score of foot-fault test improved more in the exercise group than in the control group 12, 18 and 24 days after CCI (F>4.793, P<0.05). Evans blue extravasa-tion was less in the exercise group than in the control group (t=-8.091, P<0.001), as well as the expression of MMP-2 (t=-13.12, P<0.001). Conclusion Early treadmill exercise can improve the motor function in rats with TBI, which may associate with inhibition of MMP-2 ex-pression to protect BBB integrity.

Objective To evaluate different surgical repairments for iatrogenic bile duct injury Methods Clinical data of 145 patients with iatrogenic bile duct injury surgically treated in our department from Mar 1990 to Sep 2002 were retrospectively analyzed Results Bile duct end to end anastomosis、 or bile duct repair using the patient′s own tissues and medical glue, or Roux en Y bile duct jejunostomy were used in these 145 cases The average follow up time was 3 6 years (3 months to 11 years) in 130 cases The eutherapeutic rate was 94 6% Conclusion With good surgical expertise iatrogenic bile duct injury could be repaired by a variety of methods with a satisfactory results

Exercise quantity is one of the important indexes used to evaluate physical consumption during training, so it is necessary to design a kind of instrument to accurately measure physical consuption. It must be practical, portable and with a low power consumption. This article introduces the mechanism of measuring exercise quantity. The methods of traditional and acceleration compared, the latter is selected. The design is described from the two aspects of general design and component choosing.

Objective To evaluate the feasibility of chest digital tomosynthesis(DTS)for lung lesion screening by comparing the effective dose of chest DTS with chest digital radiography(DR),low-dose MSCT and MSCT examinations.Methods The Fluke lung/chest phantom underwent posterior-anterior (PA),left lateral(LAT)chest DR and DTS with automatic exposure control technique.Using RTI DoseGuard and WinODS,the dose area product(DAP)and effective dose of PA,LAT and total DTS were calculated.CareDose technique was used for MSCT and low-dose MSCT scans,the dose length products (DLP)was acquired.According to the DLP to E(k)conversion coefficient in ICRP 103,the effective dose of low-dose MSCT and MSCT were calculated.Paired t test was used for comparison of the mean effective dose of DTS,DR and low-dose MSCT.Results The mean effective dose was 0.1 3 mSv for chest DR and 0.11 mSv for DTS examination.The mean effective dose of low-dose MSCT and MSCT scans were 1.13 mSv and 6.38 mSv.The effective dose of chest DTS was comparable to that of chest DR,and was approximately 1/10 and 1/60 times lower than that of low-dose MSCT and MSCT scans.There was no statistical difference between chest DTS and DR(t =3.514,P ＞0.01),and there was a significant difference between chest DTS and low-dose MSCT(t =178.769,P ＜0.01).Conclusion DTS is a new X-ray tomography which has the advantage of low radiation dosage in chest examination for lung lesion screening comparing with low-dose MSCT.

The object of this study is to preparate the berberine hydrochloride (BBH) resin compound with taste masking effect. We took the BBH as the model drug and Amberlite IRP69 as the drug carriers, uncovered the curve of solubility of BBH in different cosolvent with a certain range of temperature, and then used it to calculate the parameters during the preparation of the complex such as adding quantity of BBH and the reaction temperature. Afterwards, the characteristic and in vitro release experiments were studied to verify the formation and predict the in vivo release behavior of the complex. The results showed that in the condition of using 60% ethanol as a cosolvent and stirring at 50 ℃ for 1 h, the drug loading and drug availability of the complex are at about 35% and 64%, respectively, and has a better taste-masking effect. In this study, a method was provided for preparing a taste-masking preparation of BBH.

Objective To compare the stability effect of the fixation segment using the new test system based robotics to simulate the complex human movement at natural state about the three-level fixa-tion by using four, five or six pedicle screws in treatment of thoracolumbar burst fractures. Methods Six human cadaveric spines were dissected from T_(11)-L_3. The inferior half part of L_1 vertebral bodies and L_1-L_2 dises were resected to mimie an unstable L_1 burst fracture with loss of anterior column support. Specimens were tested in accordance with the following order:intact, 4,5 and 6 pedicle screws fixation at the three-level fixation. The range of motion (ROM) of the fixation segment (T_(11)-L_3) was measured with the six-freedom degree robotics system controlled by mixed force and displacement during flexion, exten-sion, lateral bending and axial torsion, when the stiffness was calculated. One-way statistieal analysis was used for analyzing the collected data. Results With increased number of screws, the ROM of the fixa-tion segment (T_(11)-L_3) was gradually decreased and the stiffness gradually increased. The ROM under ax-ial rotation of six and five screws group became smaller than four screws group (P < 0.05). The stiffness under axial rotation of six screws group was higher than four screws group (P < 0.05). There was no sta-tistical difference between five screws group and four screws group in regard of the stiffness in axial rota-tion (P > 0.05). There were no significant differences in ROM and stiffness under six different loading directions between six screws group and five screws group (P > 0.05). No statistical difference was observed for three fixation modes in aspects of ROMs and stiffness under flexion-extension or lateral ben-ding (P > 0.05). Conclusions Three-level fixation of burst fractures with five or six screws offers im-proved biomechanical stability compared with traditional four screws fixation. But the difference is insig-nificant between six and five screws fixations.

OBJECTIVETo observe the effect of nourishing Yin, strengthening Qi and activating blood decoction on Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression.

METHODThirty-two NOD mice were randomly divided into the model group, the traditional Chinese medicine group (TCM group, orally given 0.4 mL nourishing Yin, strengthening Qi and activating blood decoction as per 100 g x kg(-1) everyday), the hydroxychloroquine group (given 0.4 mL hydroxychloroquine as per 60 mg x kg(-1) everyday), the traditional Chinese medicine and western medicine group (TCM WM group, given nourishing Yin, Strengthening Qi and activating blood decoction 50 g x kg(-1) and hydroxychloroquine 60 mg x kg(-1), 0.4 mL everyday), with eight mice in each group. Eight Balb/C mice were selected as the normal control group (normal group). All of mice were killed after eight weeks, and their submaxillary glands were dissected. The expression levels of Fas/FasL were examined by immunohistochemical method, and the FasL mRNA was detected by RT-PCR.

RESULTThe expression levels of Fas/FasL in salivary glands of the model group were higher than that of other groups (P < 0.05). The expression level of FasL of the normal group was much lower than that in the hydroxychloroquine group (P < 0.05). The relative expression level of Fas mRNA in salivary glands of the model group was higher than that in other groups, but the control group was notably lower than other groups (P < 0.05). The expression level of FasL mRNA in salivary glands of the model group was higher than that in TCM and TCM WM groups (P < 0.05). But the expression level in TCM WM group was notably lower than the hydroxychloroquine group (P < 0.05).

CONCLUSIONThe nourishing Yin, strengthening Qi and activating blood decoction could down-regulate the expression level of Fas/FasL in salivary glands of NOD mice with Sjogren's syndrome and their mRNA expression, and had a better efficacy after being combined with hydroxychloroquine. The nourishing Yin, strengthening Qi and activating blood decoction might treat the Sjogren's Syndrome by reducing apoptosis which is regulated by Fas/FasL

Animals ; Fas Ligand Protein ; genetics ; Female ; Gene Expression Regulation ; Medicine, Chinese Traditional ; methods ; Mice ; Mice, Inbred NOD ; Qi ; RNA, Messenger ; genetics ; metabolism ; Salivary Glands ; metabolism ; Sjogren's Syndrome ; blood ; genetics ; therapy ; Yin-Yang ; fas Receptor ; genetics

Abstract: At present, nucleic acid detection technology based on the PCR principle is commonly used to detect malaria parasites, the existing Plasmodium detection methods mainly include microscopy, antigen immunoassay, and nucleic acid detection,but due to the long detection time, high personnel and equipment requirements, and other shortcomings, its popularization, and application at the grassroots level are limited. What challenges previous Plasmodium detection methods are the lack of experienced professionals and advanced equipment at the grassroots as well as the requirement of rapid detection of large samples under extreme conditions. The isothermal amplification technology developed in recent years has potential application prospects due to its simplicity, rapidity, high sensitivity, and high specificity. This article attempts to review the principles, characteristics, and prospects of various isothermal amplification technologies, and on this basis, focuses on the introduction of recombinase polymerase amplification (RPA) and recombinase⁃aided isothermal amplification (RAA) assay technologies and proposes the use of such recombinant enzyme amplification technologies to achieve rapid and accurate diagnosis of common Plasmodium species possibility and imagination.