Objective To construct and identify the siRNA eukaryotic expression vector targeting gene CXC chemo-kine receptor-4 and explore its role in invasion process of breast cancer cells in vitro. Methods Two siRNAs were designed and synthesized according to the coding sequence of CXCR4 gene and cloned into eukaryotic expression plasmid pGE-1-U6/kna. The constructed CXCR4-siRNA expression vector was transfected into MDA-MB-231 cells by liposome. Western blot was used to evaluate the suppression of CXCR4 expression in different groups. The inva-sion and migration of MDA-MB-231 cells were evaluated by cell invasion assay in vitro. Results Enzyme digestion and DNA sequencing confirmed that the CXCR4-siRNA expression vector was successfully constructed. After trans-fection,the CXCR4-siRNA obviously suppressed the expression of CXCR4 compared with control groups and the ability of cell migration was decreased markedly. Conclusion CXCR4-siRNA expression vector can effectively sup-press CXCR4 expression in the breast cancer cells and decrease potential of cell invasion, which may provide a no-vel strategy for gene therapy of breast cancer metastasis.

Objective To investigate the protective effects of midazolam on noise-induced hearing loss in guinea pigs by testing reactive oxygen species (ROS) level in the cochlea and plasma SOD and MDA. Methods Totally forty male pigmented guinea pigs were randomly divided into four groups: control (C) , midazolam (M), normal saline (S) and noise-induced deafened (D) groups, with ten guinea pigs in each. Groups M, S and D were exposed to a continuous noise (4kHz , octave band, 100dB SPL) 3h every day for 3 consecutive days. Group M was treated with midazolam, which was administered intramuscularly (0.1mg/kg) 24h before noise exposure, and immediately upon and during noise exposure. Group S was exposed to noise and treated with the same volume of normal saline intramuscularly, the time of injection was the same as that of Group M. Group C was not exposed to noise, but was treated with midazolam intramuscularly, the time of injection and the dosage were the same as those of Group M. Group S was exposed to noise and treated with normal saline intramuscularly ,the time of injection was same with that of Group M.Group D was exposed to noise only. All animals received auditory brainstem response (ABR) threshold recording before and immediately after noise exposure. Blood was collected when the guinea pigs were killed after the last ABR threshold recording, and serum SOD activity and MDA content were detected. Both the cochleae were removed and prepared for ROS assay. Results After noise exposure, ABR threshold shift (1.6±1.5) and ROS content [(291.10±2.30)u/mL] in Group M were significantly lower than those in Groups S and D [41.7±3.3, 44.3±3.9; (348.52±3.60)u/mL, (315.56±6.70)u/mL, P<0.05]. Serum SOD activity and MDA content were significantly increased in Group M, but the amplitude was less than that in Groups S and D.Conclusion Midazolam can prevent noise-induced hearing loss by reducing the increased ROS level in the cochlea after noise exposure.

Objective To construct and identify the siRNA eukaryotic expression vector targeting gene CXC chemokine receptor-4 and explore its role in invasion process of breast cancer cells in vitro.Methods Two siRNAs were designed and synthesized according to the coding sequence of CXCR4 gene and cloned into eukaryotic expression plasmid pGE-1-U6/kna.The constructed CXCR4-siRNA expression vector was transfected into MDA-MB-231 cells by liposome.Western blot was used to evaluate the suppression of CXCR4 expression in different groups.The invasion and migration of MDA-MB-231 cells were evaluated by cell invasion assay in vitro.Results Enzyme digestion and DNA sequencing confirmed that the CXCR4-siRNA expression vector was successfully constructed.After transfection,the CXCR4-siRNA obviously suppressed the expression of CXCR4 compared with control groups and the ability of cell migration was decreased markedly.Conclusion CXCR4-siRNA expression vector can effectively suppress CXCR4 expression in the breast cancer cells and decrease potential of cell invasion,which may provide a novel strategy for gene therapy of breast cancer metastasis.

Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.

Objective To investigate the efficacy and safety of the combined therapy of cyclophosphamide and thalidomide in the treatment of refractory Crohn's disease (CD).Methods This study was a prospective and open study.A total of 15 patients with refractory CD were enrolled.All patients received intravenous cyclophosphamide 200 mg every other day for two weeks,then followed by intravenous 400 mg once a week until the cumulative dose reached 6 to 8 g.when the cyclophosphamide treatment started,at the same time thalidomide was taken 25 to 75 mg every night according to the tolerance of patients.Before the treatment,two weeks' after the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g,Crohn's disease activity index (CDAI),hemoglobin (Hb),white blood cell (WBC) count,erythrocyte sedimentation rate (ESR) and high-sensitivity C-reactive protein (hs-CRP) were recorded.Endoscopy examination was conducted before the treatment and at the time when the cumulative dose of cyclophosphamide reached 6 to 8 g.The condition of mucosa healing was observed and scored by simple endoscopic score for crohn's disease (SES-CD).Adverse effects of all patients were monitored.Paired t test was performed for statistical analysis.Results Before the treatment,the CDAI of 15 patients with refractory CD was 235.87±59.87,two weeks after the treatment the CDAI declined to 135.33 ± 29.23,and the difference was statistically significant (t=7.50,P＜0.01).Before the treatment,ESR and hs-CRP was (42.13±22.80) mm/1 h and (13.73± 2.18) mg/L.Two weeks after treatment they declined to (23.80±16.63) mm/1 h and (5.77±4.77) mg/L,and the differences were statistically significant (t=2.43 and 6.17,both P＜0.05).After two-week treatment,10 patients achieved clinical remission.After the cumulative dose of cyclophosphamide reached 6 to 8 g combined therapy,CDAI of patients was 108.14 ± 47.10,which decreased significantly compared with that before treatment (t=6.30,P＜0.01).ESR,hs-CRP and WBC count was (19.35± 19.18) mm/1 h,(6.16± 5.02) mg/L and (6.28 ± 3.42) × 109/L,respectively,which decreased compared with those before treatment,and the differences were statistically significant (t=5.90,5.40 and 3.71,all P＜0.01).Twelve patients achieved clinical remission.And the lesions of 12 patients improved under endoscope,furthermore,the mucosa of four patients healed.Before the treatment,SES-CD was 9.14 ± 5.39,which declined to 5.07 ± 4.58 after the treatment,and the difference was statistically significant (t =3.14,P ＜ 0.01).During the treatment,five patients had adverse effects.Alanine aminotransferases (ALT) increased in three patients,WBC count decreased in one patient and one patient got a severe urinary infection.Conclusions Patients with refractory CD could achieve clinical remission,mucosa healing under endoscopy and better efficacy with the combined therapy of cyclophosphamide and thalidomide.However,adverse effects should be monitored during the treatment.

Objective To explore the clinical significance of preoperative ultrasound-assisted drawings in location of thyroid micronodule resection .Methods A total of 88 patients ( 137 nodules ) who underwent thyroid micro-nodule resection were enrolled in the prospective randomized controlled study .All patients were randomly divided into two groups:46 patients (68 micronodules) in experimental group and 42 patients (69 micronodules) in control group.Inclusion criteria:the maximum diameter of nodule≤1.0 cm. Preoperative thyroid ultrasound was conducted .Patients in experimental group also underwent ultrasound-assisted location of thyroid micronodule .Results The diagnostic accuracy rate of US was 81.82%(69/88) in all patients .With the help of ultrasound-assisted location , all nodules in experimental group were found quickly and accurately.The resection rate of experimental group was 100%(46/46).Whereas 4 nodules (in 4 patients ) were missed in control group with a resection rate of 90.5% (38/42).The postoperative US examinations after 3 months showed that all nodules were completely removed in experimental group while 4 nodules retained in control group .Conclusions Preoperative ultrasound-assisted drawings in location of thyroid micronodule plays an important role in thyroid nodule resection .It is great value of clinical application .

Objective To evaluate the accuracy of renal scintigraphy for the estimation of glomerular filtration rates (dGFR) in patients with diabetic nephropathy as compared to the conventional dual-plasma sample clearance method (pscGFR). Methods Forty-six patients with diabetic nephropathy underwent both dynamic renal scintigraphy and dual-plasma sample measurement after 99Tcm-DTPA injection. Paired student t-test and correlation analysis were performed to compare dGFR and pscGFR (normalized to body surface area,1.73 m-2). Results The mean dGFR was higher than mean pscGFR ((51.08±26.78)ml·min-1vs (44.06±29.43)ml·min-1,t=4.209,P=0.000). The dGFR correlated with pscGFR ( r=0.923,P=0.000) linearly (regression equation:pscGFR=1.015×dGFR-7.773,F=254. 656,P=0.000).Conclusions dGFR correlated well with pscGFR. Although it could not absolutely replace the latter in patients with diabetic nephropathy,dGFR could reasonably evaluate the filtration function for these patients.

Objective Establishing an employee satisfaction indicator system for tertiary hospitals in Beijing.Methods Evaluation indicators are built on theory research,and questionnaires are designed for survey.Such methods as principal component factor analysis,and verification factor analysis are called into play to test the construct validity and establish in the end the employee satisfaction indicators system.Results Such indicators comprise five grade-1 dimension and 34 grade-Ⅱ dimension,such as hospital management satisfaction indicator.Conclusion This self-designed scale ideally mirrors the employee satisfaction of tertiary hospitals in Beijing,proving the indicators to be objective and reasonable.

Objective:To determine the effect of Chuju total flavonoids (CJTF) on the secreted frizzled-related protein 4 (SFRP4) expression in Wnt pathway in rheumatoid arthritis (AR) model rats. Methods:hTe role of CJTF in the treatment of AR model rats was evaluated by rat arthritis score and paw edema score. The expression regulation of the SFRP4,β-catenin and C-myc in Wnt pathway in AR model rats was detected by RT-PCR and Western blot atfer CJTF gavage treatment. Results:Atfer CJTF treatment, the rat arthritis score and paw edema score in AR model rats were signiifcantly decreased when the AR model rats were treated with CJTF, the SFRP4 expression was signiifcantly up-regulated, while theβ-catenin and C-myc gene expression were signiifcantly down-regulated in AR model rat synovial tissues. Conclusion:CJTF has significant therapeutic effect and inhibitory effect on Wnt pathway activation by targeting SFRP4 in AR model rat synovium.

BACKGROUND: Thoracolumbar burst fracture Dennis type B does not have significant damage in the intervertebral discs of the inferior vertebral body. To reduce fusion segment and remain normal intercalated disc, single discectomy (damaged vertebral body and supervisor vertebral body) is proposed, but because of the damage to the vertebral body, implanted screw is easy to loose following excising partial vertebral body, even cannot be fixed. However, it is still unknown whether implanted screw in the inferior vertebral body of the damaged vertebral body was stabilized using two segment discectomy with fixation.OBJECTIVE: To analyze the feasibility of single segmental spinal interbody fusion with bisegment fixation for thoracolumbar burst fracture type B.DESIGN, TIME AND SETTING: The randomized controlled in vitro study was conducted at the Laboratory of Biomechanics, Southern Medical University from June 2007 to June 2008.MATERIALS: A total of 20 fresh freezing 7-9 months pig samples (T_(13)?L_3 segment) were used, comprising 10 integral samples and 10 L_1 type B thoracolumbar burst fracture samples prepared by pre-injury and weight dropping technique.METHODS: Pig fresh thoracolumbar specimens from T_(13)?L_3 were collected to create models of type B thoracolumbar burst fracture. There were 4 groups in this study. Ten of them were selected as intact group (n=10) (fresh pig T_(13)?L_3 segment). T_1 vertebral endplate pre-injury and weight dropping technique and incremental trauma approach were used. Denis' type B burst fracture was produced, and ten of them were selected as unstable group (n=10). Firstly, unstable group was decompressed by discectomy and semivertebraectomy in upper half of the vertebral body, single level was fused with iliac and U-FRONT anterior thoracolumbar system were placed between T_(14) and L_2, as single discectomy with fixation group (n=10). Then lower disc of injury vertebra discectomy and vertebraectomy, fused with iliac U-FRONT anterior thoracolumbar system were placed between T_(14) and L_2, as two segment discectomy with fixation group (n=10). The bone graft was longer 1 mm than the bone graft region.MAIN OUTCOME MEASURES: The flexion, extension, right/left lateral bending, and right/left axial rotation range of motion (ROM) of T_(14)?L_2 were measured in each group on the spinal three-motional test machine at 10 N穖.RESULTS: The flexion, extension, right/left lateral bending, and right/left axial rotation were not stable in the unstable group.ROM was significantly increased in the unstable group compared with the intact group (P< 0.01). The primary stability was significantly elevated in the single discectomy with fixation and two segment discectomy with fixation groups. The flexion, extension, right/left lateral bending, and right/left axial rotation ROM were significantly reduced in the single discectomy with fixation and two segment discectomy with fixation groups compared with the unstable group (P < 0.05). The flexion, extension, right/left lateral bending ROM was significantly decreased in the two groups compared with the intact group, but axial rotation ROM was significantly increased (P< 0.05). Axial rotation ROM was smaller in the single discectomy with fixation group compared with the two segment discectomy with fixation group (P < 0.05).CONCLUSION: Single segmental spinal interbody fusion with bisegment fixation for thoracolumbar burst fracture type B had a good immediate stability in flexion, extension, lateral bending motion. Compared with traditional partial corpectomy L_1 between the caudal and cranial endplate of the adjacent vertebrae with bisegmental fixation, it had a better immediate stability in axial rotation.