BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy.
OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro.
METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal
interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours.
RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (P<0.05), and the inhibition effect was more significant at 72
hours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ,
feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after
trasfection (P<0.05), and the inhibition effect was more significant at 72 hours after transfection when compared with that at 48 hours after transfection, which was same with the effect on mRNA level. RNA interference in vitro can
significantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.

By the bioactivity technology of Low-Temperature Preparation of Anatase and Rutile Layers on porous Titanium Substrates, CPTi(Commercially pure titanium Phi4.0 mm x 20 mm) was processed and treated as experiment specimens while CPTi was treated as control specimens. Experiment specimens and control specimens were implanted into the holes (4.0 mm in diameter) in rabbit's right and left tibias respectively. After implantation for predetermined periods of 4,8,12, 16 weeks, the specimens were taken out with bone tissues, and were examined by a press-out tester to evaluate the shearing force between the implant and the bone tissue. It is found that the shearing force between the experiment specimen and the bone is more significantly higher than that between the control specimen and the bone, and the bonding time is shorter, the stabilization time is faster. This study has laid down a good foundation for the titanium metal's innovative applications in clinical orthopaedics.
Animals ; Bone Substitutes ; chemistry ; Bone and Bones ; Cold Temperature ; Crystallization ; Materials Testing ; Orthopedic Fixation Devices ; Prostheses and Implants ; Rabbits ; Tibia ; Titanium ; chemistry

OBJECTIVETo optimize the adsorption condition of cation-exchange chromatographic media Streamline SP for separation and purification of anti-HBsAg Fab fragment from E. coli.

METHODSThe adsorption of the target protein for separation and purification by the cation-exchange chromatographic media Streamline SP was tested using test tube method in balanced buffer solution with different pH values and ion concentrations. The adsorption effect was then verified by cation-exchange chromatography using 1-ml Streamline SP prepacked column and 28-ml Streamline SP self-assembly column.

RESULTSAccording to the experiment results of test tube method, the loading buffer with pH of 4.4 and ionic concentration of 100 to 600 mmol/L could achieve optimal target protein adsorption effect by cation-exchange chromatographic media Streamline SP, as verified by cation-exchange chromatography with 1-ml SP prepacked column and 28-ml Streamline SP self-assembly column.

CONCLUSIONThe optimal condition of cation-exchange chromatography selected by test tube method can be applied for separation and purification of anti-HBsAg Fab fragment from E. coli.

Adsorption ; Cation Exchange Resins ; Chromatography, Ion Exchange ; methods ; Escherichia coli ; genetics ; metabolism ; Hepatitis B Antibodies ; isolation & purification ; metabolism ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin Fab Fragments ; isolation & purification ; metabolism

OBJECTIVETo explore the effects of bevacizumab with or without cisplatin (DDP) on the growth of lung adenocarcinoma A549/DDP cell xenografts in mice.

METHODSHuman lung cancer A549/DDP cells was subcutaneously transplanted in to 25 nude mice, which were randomly divided into control group (group A), bevacizumab group (group B), DDP group (group C), combined treatment group (group D) and half-dose combined treatment group (group E). After corresponding treatments for 4 consecutive weeks, the tumor inhibition rate was evaluated, tumor microvessel density (MVD) measured with immunohistochemistry, and the mRNA expression of apoptosis-associated gene (bcl-2) and multidrug resistance genes (LRP and GST-pi) assessed by RT-PCR.

RESULTSThe tumor growth inhibition rates in groups B, D, and E with bevacizumab treatment were 20.96%, 51.67% and 50.95%, respectively, and the two combined treatment groups showed better effects. MVD in these 3 groups were 18.6-/+1.14, 13.6-/+1.14, and 14.4-/+0.55, respectively, and no significant difference was found in MVD between DDP group and the control group. Compared with the control group, the 3 bevacizumab-treated groups showed decreased expression of bcl-2 genes in A549/DDP tumors at a comparable amplitude, and LRP and GST-pi mRNA expression showed no significant differences between the 5 groups.

CONCLUSIONBevacizumab has synergetic inhibitory effect with conventional chemotherapy against lung adenocarcinoma A549/DDP cell xenografts in mice by inhibiting angiogenesis of the tumor, and may enhance the sensitivity of A549/DDP cells to DDP by inducing cell apoptosis.

Adenocarcinoma ; genetics ; pathology ; Animals ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Bevacizumab ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; Cisplatin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Mice ; Mice, Nude ; Xenograft Model Antitumor Assays

OBJECTIVETo explore the discrepancy proteins in gastric cancer by proteome analysis.

METHODSTotal proteins of gastric cancer tissues and matched normal gastric epithelial tissues were separated respectively by two-dimensional polyacrylamide gel electrophoresis (2-DE). Mass spectrometry was used to test the differentially expressed proteins.

RESULTSOne thousand one hundred and forty-seven protein spots from gastric cancer tissue and 1079 spots from the normal tissue were gained. Out of 164 different protein spots, 41 were only expressed in gastric cancer tissue, 27 were unique in normal tissue, 39 were up-regulated and 57 were down-regulated in gastric cancer. Seven proteins, which were highly expressed in gastric cancer tissue, were identified.

CONCLUSIONDifferent protein spots between gastric cancer tissues and normal gastric epithelial tissue were gained by proteomics. The 7 discrepancy proteins were further identified. It establishes the foundation of finding specific gastric cancer proteins, which act as biomarkers for the diagnosis and prognosis of gastric cancer.

Case-Control Studies ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Mass Spectrometry ; Peptide Mapping ; Proteomics ; methods ; Stomach Neoplasms ; metabolism

OBJECTIVESTo evaluate the relationship between Modic change and disc height together with lumbar hyperosteogeny and study the role of Modic change in lumbar degeneration.

METHODSThe imaging data of 150 elderly patients with chronic low back pain were analysed retrospectively. All patients underwent MRI and lumbar lateral X-ray examination. The lumbar disc from L1-L2 to L5-S1 were selected for this study, including 750 discs, vertebral and endplate close to disc in 150 patients. The incidence rate of lumbar endplate Modic change, disc height and the degree of vertebral bone hyperplasia were recorded. The ratio of disc height/lumbar intervertebral disc height < 50% was defined as disc collapse. The patients were divided into 4 groups in the basis of imaging changes. Group A1:disc collapse without severe lumbar hyperosteogeny; Group A2: disc collapse with severe lumbar hyperosteogeny; Group B1: Neither disc collapse nor severe lumbar hyperosteogeny; Group B2: severe lumbar hyperosteogeny without disc collapse. The incidence rates of Modic change were compared between the 4 groups by χ(2) test. Finally, the influence of disc height and vertebral bone hyperplasia on the incidence rate of Modic change was analysed.

RESULTSFour groups of patients observed a total of 750 discs. The number of intervertebral discs in the group A1 was 208, the incidence rate was 54.3%. The number of intervertebral discs in the group A2 was 135, the incidence rate of group A2 was 34.8%. The number of intervertebral discs in the B1 group was 225, the incidence rate of group B1 was 16.9%. The number of intervertebral discs in the B2 group was 182, the incidence rate of group B2 was 29.7%. There was significant difference of lumbar endplate Modic change incidence rate among the 4 groups(χ(2) = 69.565, P < 0.05). The results of post hoc test showed that the incidence rate of Modic change in group A1 was higher than group A2, B1 and B2 (χ(2) = 12.524, 66.701 and 24.102, P < 0.00714). There was significant difference of Modic change incidence rate between group A2 and B1(χ(2) = 15.032, P < 0.00714), but there was no significant difference of Modic change incidence rate between group A2 and B2 (χ(2) = 0.945, P > 0.00714) . There was significant difference of Modic change incidence rate between group B2 and group B1 (χ(2) = 9.395, P < 0.00714).

CONCLUSIONSThe incidence rate of Modic change with disc collapse but without severe lumbar hyperosteogeny is high in elderly patients with chronic low back pain. There is no significant difference of Modic change incidence between patients with both disc collapse and severe lumbar hyperosteogeny and patients with severe lumbar hyperosteogeny but without disc collapse.

Aged ; Aged, 80 and over ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Degeneration ; pathology ; Low Back Pain ; pathology ; Lumbar Vertebrae ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies

A reasonable method for the quality control of tablets of Ginkgo biloba leaves was established in this paper. The total flavonol glycosides and terpene lactones of G. biloba tablets were quantified by HPLC. Totally, 16 batches of the commercially available tablets of G. biloba leaves were determined. Among of them, 2 batches were unqualified in the content of total flavonol glycosides, and 3 batches were unqualified in the content of terpene lactones. A validated HPLC fingerprint method was established to evaluate the commercially available tablets of G. biloba leaves with the assistance of LC-MS. Sixteen batches showed the similarity of 0.763-0.989. There were 31 fingerprint chromatogram peaks were identified as flavonoids compositions by LC-MS. This provides a research idea for the quality control of tablets of G. biloba leaves.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Ginkgo biloba ; chemistry ; Mass Spectrometry ; methods ; Plant Leaves ; chemistry ; Quality Control ; Tablets ; chemistry

OBJECTIVETo investigate the pollution level and development trend of lead in the preserved egg in our country.

METHODSBy the national food contamination monitoring system and under the strict analysis quality control, the content of lead in the preserved eggs was analyzed according to the national standard method (GB/T 5009.12-2003) in fourteen provinces from 2000 to 2006.

RESULTSAll 1358 data on contents of lead in the preserved eggs were obtained during seven years, the total average was 1.782 mg/kg, the maximum was 334.0 mg/kg, P90 was 3.50 mg/kg, P95 was 7.397 mg/kg and P97.5 was 12.01 mg/kg, all exceeded 2 mg/kg of the national limit standard, and the rate of violated samples exceeded 10.0%. Analyzing from time, contents of lead in the preserved eggs were depressive from 2.994 mg/kg to 1.138 mg/kg year after year.

CONCLUSIONThe lead contamination in preserved eggs was serious in whole country. It shows that the continuous work of monitoring and forewarning should be carried out to make the contamination of lead in preserved eggs to reduce year by year.

China ; Eggs ; analysis ; Food Contamination ; analysis ; Food Handling ; methods ; Food Inspection ; methods ; Lead ; analysis

OBJECTIVETo test whether the tumor vessel density (TVD) from the enhanced spiral CT can preoperatively predict nodal status and distant metastasis of colorectal cancer.

METHODSForty cases of colorectal cancer patients who received surgical treatment were included in this study. The three dimensional tumor vessels were reconstructed by an enhanced CT 64-slice spiral CT and its AW4.4 image processing platform. The TVD was measured by the 1000 high-resolution color graphics pathological analysis system. The TVD level was compared between different tumor size, classification, and TNM stage. The postoperative pathological staging was taken as golden standard.

RESULTSThe sensitivity, specificity and accuracy for direct prediction of lymph node metastasis by the enhanced CT 64-slice spiral CT was 74.1%(20/27), 53.8%(7/13) and 67.5%(27/40) respectively. The TVD from the reconstructed three dimensional tumor vessels in the group with lymph node metastasis was significantly higher than that without metastasis(0.070±0.046 vs. 0.037±0.013, P<0.05). The TVD in the distant metastasis group was significantly higher than that without distant metastasis (0.130±0.032 vs. 0.049±0.030, P<0.01). No difference of TVD was found between different tumor size, invasion depth, and differentiation type.

CONCLUSIONTVD level from the reconstructed three dimensional tumor vessels can indicate lymph node and distant metastasis of colorectal cancer.

Colorectal Neoplasms ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Neoplasm Staging ; Tomography, Spiral Computed

OBJECTIVETo study the etiology and clinical significance of axial symptoms after posterior operative procedures for ossification of the posterior longitudinal ligament (OPLL).

METHODSFrom February 2005 to February 2010, 76 patients with OPLL treated were retrospectively experienced. There were 34 male and 42 female with average of 52.1 years (range from 37 to 74 years), the average duration of the disease was 32.1 months (range from 11 to 56 months). Nineteen patients underwent traditional laminectomy in group A, 33 patients received open-door laminoplasty in group B and 24 patients underwent lateral mass screw fixation in group C. All patients underwent X-ray examination pre- and post operative, computed tomography were used for diagnosis of OPLL, the recovery rate was calculated using pre- and postoperative Japanese Orthopedic Association (JOA) scores for each patient. Pre- and postoperative cervical curvature index and axial symptoms were measured and compared. χ(2) test and SNK test were used as statistical methods.

RESULTSAll patients were followed up for 14 - 35 months, average (21 ± 5) months. Loss of cervical curvature index was 4.2% ± 1.7% in group A, 2.9% ± 2.2% in group B and 2.3% ± 1.9% in group C. The difference was significant in loss of cervical curvature indice between group A and B (q = 2.94, P < 0.01), group A and C (q = 4.23, P < 0.01). The average JOA recovery rate was 58.3% for group A, 64.3% for group B and 66.7% for group C. There was no significant difference in JOA recovery rate among the three groups (P > 0.05). The rate of early evident axial symptoms was 7/19 in group A, 30.3% in group B and 33.3% in group C and the difference was not statistically significant (P > 0.05). The incidence of late evident axial symptoms was 5/19 in group A, 12.1% in group B and 8.3% in group C, the difference was not significant between group B and C (χ(2) = 13.762, P < 0.01), but of statistical difference between group A and B(χ(2) = 6.368, P < 0.01), group A and C (χ(2) = 11.481, P < 0.01). No kyphotic deformity in the group A, no "Close Door" phenomenon in group B and no internal failure in group C.

CONCLUSIONThe incidence of early axial symptoms are of no significant difference among the three groups, but late axial symptoms are higher in the laminectomy than other groups, which may be associated with loss of cervical lordosis.

Adult ; Decompression, Surgical ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Laminectomy ; adverse effects ; methods ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Postoperative Complications ; Retrospective Studies