Objective To construct small RNAi plasmid vectors which can suppress the expression of TITIN in Wistar rat. Methods A recombinant plasmid vector involving shRNA, which matched the base pair of rat TITIN N2B region mRNA perfectly, was constructed and transfected into normal neonatal cardiomyocytes and mesenchymal stem cells (MSCs) induced by 5-aza cytidine respectively. The expression of TITIN Z band was investigated by immunofluorescence. Results The expression of TITIN was weakened after recombinant plasmid was transfected into neonatal cardiomyocytes and MSCs induced by 5-aza cytidine. Conclusion The recombinant plasmid was constructed successfully, and it actually can block the expression of TITIN.

Objective:A database was designed and developed for effective utilization monitoring data of radiation dose to patients and occupational operators in cardiovascular interventional procedures. Methods:To complete a command reception of the client program, data display and other functions, compiled database software using MS Visual Basic level programming language. Using MS Access database completed data storage, management and maintenance. Results:The database was designed according to technological indicator acknowledged;the relevant help file was set up. The database includes two sub-libraries professional database and patient database. Professionals and patients can be recorded separately input, record view and record query. Conclusion:The data platform for monitoring data of radiation dose in cardiovascular interventional procedures was provided, which can provide basic data for constituting normalization and technical standard of dose limited of radiation dose to patients and occupational operators in cardiovascular interventional procedures.

The social reputation-based incentive mechanisms in preventing the free riding behaviors were put for-ward according to the analysis of personal digital library and P2P technology. Since the reputation value of personal digital library was calculated according to the maximum flow algorithm, personal digital library must increase its reputation value by sharing the resources when it is going to obtain the necessary resources it wants.

Objective To explore the technique outcome of fixing the ventral stress pedicle screws into the injured vertebrae, as a method to enhance the posterior internal fixation. Methods From March 2002 to March 2005, 33 single thoracolumbar fractures were studied retrospectively. Among which, 16 cases were treated with the above method(group A), and the other 17 were treated with traditional two-level fixation(group B). Group A involved 11 males and 5 females, aged 48 years averagely(range, 32-74 years); and group B included 12 males and 5 females, aged from 21 to 61 years(mean, 40 years). All the patients underwent the operation within up to 3 weeks after fracture. For the injured vertebral bodies, their pedicles were intact on either unilateral or bilateral side, and their lower half and endplate were free from split. In group A, the pedicle screws in the injured vertebrae were used to achieve the ventral press vertical to the distraction for the stress neutralization, and also with the routine distraction and lordosis restoration, simultaneously. The mean follow-up period was 11 months with a range from 6 to 24 months. Results After operation, the optimal Cobb angle and anterior column restoration were achieved through the ventral reduction from the injured vertebral body, which was the contribution from the vertical stress pedicle screw. The degree in anterior movement of injured vertebrae pre- and postoperatively was much more in group A than group B, and the difference was of statistical significance(P

[Objective]To investigate the efficacy of treatment of C_(1～C) instability with 3D-fixation techniques.[Method]From July 2002 to July 2005,thirteen patients were treated with atlantal lateral mass screw associating with axoid pedicle screw;twelve patients were treated with axoid transarticular screw associating with posterior atlantal arch hook;sixteen patients were treated with Vertex and nine patients with Summit among them.[Result]There were 26 lateral mass screws,26 axoid transarticular screws,24 atlantoaxial transarticular screws and 24 posterior atlantal arch hooks were applied in the 25 patients.All patients were followed up.The follow up duration time was ranging from 10 to 36 months with the mean time of 16.7 months.No spinal injury and vertical artery injury was found. The postoperative JOA score was ranging from 13.6 to 15.9 with the mean score of 14.8 and the improvement rate of 89.5%.All fractures of odontoid process and all bone grafts had solid bone union.No failure and loosening of internal instrument was found.[Conclusion]The two 3D-fixation techniques are effective and reliable.

OBJECTIVE:To study the effects and mechanism of rapamycin on invasion and metastasis of cervical cancer HeLa cell. METHODS:HeLa cells were divided into control group and rapamycin low-dose,medium-dose and high-dose groups (10, 30,100 nmol/L). After treated for 48 h,cell viability was measured by MTT assay,and inhibitory rate was calculated;migration and invasion of cell was tested by Transwell assay. The expression of matrix metalloproteinase 2(MMP-2),MMP-9,Vimentin and E-cadherin,and phosphorylation of protein kinase B (Akt),mammalian target of rapamycin (mTOR) were detected by Western blot. RESULTS:Compared with control group,the inhibition rate of cell viability was increased in rapamycin groups(P<0.01);the number of invasion and metastasis cells decreased(P<0.01);the expression of MMP-2,MMP-9 and Vimentin were decreased (P<0.01 or P<0.05);the expression of E-cadherin was enhanced(P<0.01 or P<0.05);the phosphorylation of Akt and mTOR were reduced (P<0.01). CONCLUSIONS:Rapamycin could inhibit invasion and metastasis of HeLa cell via Akt/mTOR signal pathway.

Objective To investigate effect of folic acid combined with xin funing on CRP, HGF, IL-2,TNF-αof patients with cervical cancer caused by human papillomavirus.Methods 80 cases of cervical cancer patients were randomly divided into control group, 40 cases in the control group were given conventional chemotherapy, 40 cases in the experimental group were on the base of the control with folic acid combined with xin funing.CRP, HGF, TNF-α, IL-2 and T lymphocyte subsets were compared before and after the treatment.Results Compared with the control group, the serum CRP, HGF and TNF-αof the experiment group were lower(P<0.05), IL-2 levels was higher (P<0.05), CD4 +and CD4 +/CD8 +level were higher(P<0.05), level of CD8 +was lower(P<0.05) and the clinical effective rate were higher(P<0.05).Conclusion Folic acid combined with Xin Funing has important significance for the treatment of patients with cervical cancer.It is speculated that the mechanism may be to reduce the level of serum CRP and HGF in patients with cervical cancer, and to increase the level of IL-2, and to regulate immune cells.

BACKGROUND:There are many multimodal analgesia schemes in perioperative period of total knee arthroplasty, but there is no ideal scheme. OBJECTIVE:To explore the effects of multi-mode and preemptive analgesia on analgesic effect after total knee arthroplasty. METHODS:120 patients with severe osteoarthritis who underwent unilateral knee arthroplasty were enroled in this study. According to different analgesic effects, the patients could be divided into four groups (n=30). In the control group, no corresponding analgesic measures were found before and during replacement. In the preemptive analgesia group, celecoxib was oraly taken before replacement. In the cocktail analgesia group, cocktail was periarticularly injected during replacement. In the multimodal combined analgesia group, celecoxib was oraly taken before replacement + cocktail was periarticularly injected during replacement. After replacement, intravenous patient-controled analgesia pump was applied in each group. Active flexion range-of-motion, visual analogue scale score in the resting and active states and knee Keen Society Score were measured at various time points after total knee arthroplasty in four groups. Adverse reactions were recorded after replacement. RESULTS AND CONCLUSION:Active flexion range-of-motion, visual analogue scale score in the resting and active states and knee Keen Society Score were better in the preemptive analgesia, cocktail analgesia and multimodal combined analgesia groups than in the control group at various time points after replacement (alP < 0.05). Above indexes were better in the multimodal combined analgesia group than in the preemptive analgesia and cocktail analgesia groups (alP < 0.05). No significant difference in above indexes was detected between the preemptive analgesia and cocktail analgesia groups (alP > 0.05). No significant difference in the number of cases affecting nausea and vomiting was detected at 2 weeks after replacement in the four groups (P > 0.05). There was no deep venous thrombosis of double lower limbs or necrosis and infection of incision. These findings suggest that the effects of preemptive analgesia before total knee arthroplasty, local injection analgesia during replacement, and the combined analgesia of intravenous patient-controled analgesia pump after replacement were ideal. Adverse reactions did not increase, and the operation was safe. At present, multimodal combined analgesia has been accepted by us, but to achieve truly painless results after total knee arthroplasty stil needs more efforts.

Objective To construct the eukaryotic plasmid expression vector mediated short hairpin RNA(shRNA) interference targeting TLR4 gene,and transfect it into pancreatic adenocarcinoma cell line PANC1,then screen stably transfected clonal cell line.Methods Three shRNA interference expression plasmid vectors targeting the TLR4 gene were constructed,named TLR4-1,TLR4-2,TLR4-3.The shRNA plasmid with highest inhibitory efficiency was selected and transfected into PANC1 cells with liposome.The silencing efficiency and transfection efficiency of TLR4-shRNA was assayed with real-time quantitative PCR and flow cytometry analysis.Monoclonal cell with stable transfection of TLR4-shRNA were selected by geneticin 418 (C418) and limiting dilution analysis.Results Transient transfection efficiency of PANC1 was (46.72 ±5.06) ％.TLR4 mRNA expressions were 0.025 ± 0.004,0.027 ± 0.003,0.019 ± 0.006in cells transfected with TLR4-1,TLR4-2,TLR4-3,respectively,which were significantly lower than that in untransfected group (0.061 ±0.018) and negative control group (0.057 ±0.015,P ＜0.05).The transfection efficiency of TLR4-3 vector in stably transfected clones [(82.79 ±8.16)％] was significantly higher than that of transient transfection (P =0.001 ).The expression of TLR4 mRNA was decreased to 0.010 ± 0.002,which was significantly lower than that of transient transfection ( P =0.001 ).The expression of TLR4 protein was (0.54±0.32) ％,which was significantly lower than that of untransfected cells [( 87.42 ± 5.00 ) ％] and that of negative control [(82.9±5.00)％,P =0.000].Conclusions Stable transfection PANC1 cell lines with TLR4 gene silencing are successfully identified.

A high-performance liquid chromatography coupled with mass spectrometry (HPLC–MS) method was established for the separation and determination of acetyl-glutamine enantiomers (acetyl-L-glutamine and acetyl-D-glutamine) simultaneously. Baseline separation was achieved on Chiralpak AD-H column (250 mm × 4.6 mm, 5 μm). n-Hexane (containing 0.1% acetic acid) and ethanol (75:25, v/v) were used as mobile phase at a flow rate of 0.6 mL/min. The detection was operated in the negative ion mode with an ESI source. [M-H]? m/z 187.0540 for enantiomers and [M-H]? m/z 179.0240 for aspirin (IS) were selected as detecting ions. The linear range of the calibration curve for each enantiomer was 0.05–40 μg/mL. The precision of this method at concentrations of 0.5–20 μg/mL was within 7.23%, and the accuracy was 99.81%–107.81%. The precision at LOQ (0.05 μg/mL) was between 16.28% and 17.56%, which was poor than that at QC levels. The average extraction recovery was higher than 85% for both enantiomers at QC levels. The pharmacokinetics of enantiomers was found to be stereoselective. There was not chiral inversion in vivo or in vitro between enantiomers.