Objective To evaluate graft isometry after anatomic single bundle reconstruction of anterior cruciate ligament (ACL) using dynamic computed tomography (CT).Methods A retrospective study was conducted of the 14 patients who had undergone single bundle ACL reconstruction from June to August 2015.They were all men,with an average age of 28.6 years (range,from 18 to 39 years).At 6 months after operation,they received dynamic CT scanning during a cycle of knee extension to flexion.3D bone models representing the knee at different flexion positions (0°,30°,60°,90°,and 120°) in each patient were reconstructed from the CT images.The grid method was used to locate the positions of the central footprints of the tibial and femoral tunnels.The lengths between the entries of the femoral and tibial tunnels were measured from each tunnel entry to reflect the graft length change.Furthermore,we measured the isometry at the over-the-top position of the femur and at the anatomic tibial position.Results All the tunnel entries were located at the central area of the ACL anatomic attachment.The reconstructed ACL was the longest when the knee was in full extension.The length was gradually shortened between the femoral and tibial tunnels during flexion of the knee from 0° to 90°.The anatomic position showed an average of 4.82 mm shortening and the over-the-top position an average of 3.28 mm shortening.The length excursion increased in early flexion from 0° to 30° (2.91 ±0.91 mm on average) and reduced in later flexion from 90° to 120° (2.98 ± 1.41 mm on average).Conclusions None of the reconstructed ACL was isometric.A graft length may be the longest when the knee is in full extension and decrease gradually during the flexion from 0° to 90° and increase gradually during the flexion of 90° to 120°.The graft should be fixed when the knee is in the flexion of 30°.

Objective To investigate the change of the content of suppressor of cytokine signaling (SOCS-1) in the liver of septic mice and its working mechanism.Methods Adopted Cecalligation and puncture (CLP) to create models of sepsis and divided randomly adult male BALB/c mice into 8 groups,including normal controlled group,sham-operated group,and the killed groups 2 hours,6 hours,12 hours,24 hours and 48 hours after operation.After extracting the RNA and protein from the liver tissue of the mouse groups,reverse transcription polymerase chain reaction (RT-PCR) was adopted to determine the relative content of SOCS-1 mRNA in the tissue,Western blot was adopted to determine the relative content of protein and the SPSS statistics software was adopted to calculate the correlation.Then observed the pathological change of liver tissues,and detected SOCS-1 protein expression by immunohistochemistry.Results After CLP suergery,the expression of SOCS-1 on gene degree in the liver and the expression of SOCS1 on protein degree in the liver increased rapidly at the 6th hour ( P ＜ 0.05 ),with the former reaching peak ( P ＜ 0.05 ) at the 24th hour and the latter remaining high all the time.There were pathological changes such as fatty degeneration and necrosis in the septic liver tissue,hepatic SOCS-1 protein expression could be detected by immunohistochemistry.Conclusions CLP induced sepsis could lead to the increase of the expression of SOCS1 in the liver.

Objective:To investigate the role of androgen receptor (AR) in CK5 +CK8 + cells isolated from prostate cancer LNCaP cells and its regulating mechanism. Methods:CK5 +CK8 + cells were isolated from LNCaP cells by using flow cytometry. Lentivirus vector carrying AR gene was transferred in CK5 +CK8 + cells. The experiments were divided into AR CK5 +CK8 + group transfering AR and V CK5 +CK8 + group transfering blank load. The expressions of AR, p-AKT and bcl-2 were tested by using Western blot assay under different concentrations of androgen (1 nmol/L and 10 nmol/L dihydrotestosterone). Methyl thiazolyl tetrazolium (MTT) assay, cell migration assay and soft agarose gel clone formation assay was used to detect the effect of AR on the biological property of CK5 +CK8 + cells. The effect of activated inhibitors such as LY 294002 (LY), γ-tocotrienol (γ-TT) and/or 5-fluorocytosine inducing AR expression (5-AZA) through AKT signal pathways on CK5 +CK8 + cells proliferation was detected by using MTT assay. Results:After AR gene was transferred into CK5 +CK8 + cells, the expression of AR was increased, while the expression of p-AKT and bcl-2 was decreased. After the treatment of 1 nmol /L dihydrotestosterone and 10 nmol/L dihydrotestosterone for 2, 4 and 6 d, the cell proliferation inhibited degree of AR CK5 +CK8 + cells was higher compared with that of V CK5 +CK8 + cells, and the difference was statistically significant (all P < 0.05). After the treatment of 1 nmol/L dihydrotestosterone and 10 nmol /L dihydrotestosterone for 3 d, the migration ability of AR CK5 +CK8 + cells was decreased compared with that of V CK5 +CK8 + cells (the number of cell migration: 54±9 vs. 113±21, 13±3 vs. 34±6), and the differences were statistically significant ( t=4.450, P<0.01; t=5.157, P<0.01).After the treatment of 1 nmol /L dihydrotestosterone and 10 nmol /L dihydrotestosterone for 3 weeks, the tumorigenic ability of AR CK5 +CK8 + cells was reduced compared with that of V CK5 +CK8 + cells (the number of clone: 39±7 vs. 105±16, 41±6 vs. 86±6), and the differences were statistically significant ( t=6.631, P<0.01; t=8.662, P<0.01). And 5 nmol /L LY + 10 nmol/L 5-AZA, 5 nmol /L LY + 5 nmol/L γ-TT, 10 nmol/L 5-AZA + 5 nmol/L γ-TT, 2.5 nmol/L LY + 5 nmol/L 5-AZA + 2.5 nmol/L γ-TT combined with 1 nmol/L dihydrotestosterone or 10 nmol/L dihydrotestosterone after the treatment of 2, 4, 6 d inhibited the proliferation of CK5 +CK8 + cells (all P < 0.05). Conclusion:AR plays an inhibitory role in CK5 +CK8 + cells isolated from prostate cancer cell line LNCaP and reduces the cell migration and tumorigenic ability through inhibiting activation of AKT-bcl-2 signal pathway.

This paper focused on factors which affected on different color of northern and southern region vinegar-processed frankincense.Meanwhile,contents of six main boswellic acids were also determined to elaborate the influence of heat in chemical components.Vinegar-processed frankincense from northern and southern region was collected.And different temperature and time were used in the processing of frankincense to receive the vinegar-processed frankincense samples.The color difference meter was utilized combining with the PCA statistic analysis method.The Zorbax ExtendC18 chromatographic column (4.6 mm × 50 mm,1.8 μm) was used with acetonitrile-0.1％ phosphoric acid as the mobile phase and gradient elution.The velocity of flow was 1 mL· min-1.The detection wavelength was 210 nm and 250 nm.The column temperature was 30℃.The results showed that the color of northern region processed frankincense was yellow or pale brown.And the southern region processed frankincense was pale brown or dark brown.It showed the difference on processed degree.The L* value of the northern processed frankincense was 75.327 to 80.746 and the L* value of southern processed was 44.321 to 49.527.The a* value of the northern processed frankincense was 5.378 to 6.502 and the a* value of southern processed was 9.423 to 9.978.There was no significant difference on b*.There were certain differences on L* and a* among vinegar-processed frankincense with the same surface color.The color parameter results of self-made vinegar-processed frankincense indicated that along with changes of processing temperature and time,the color,L* and a* change.Even frankincense processed for 30 min with mild fire,it will not achieve the color parameter value of the southern region vinegar-processed frankincense.However,after 11 min processing with medium fire,the color can be achieved.The content determination results showed that four contents,including α-boswellic acid,β-boswellic acid,3-acetyl-α-boswellic acid and 3-acetyl-β-boswellic acid were increased.Contents of 11-carbonyl-3-boswellic acid and 3-acetyl-11-carbonyl-β-boswellic were decreased after being processed.The range of increasing or decreasing by medium fire was higher than mild fire.At the same temperature,as the increasing of processing time,the content has an increasing or decreasing tendency.It was concluded that temperature was the main factor influencing the color of vinegar-processed frankincense from northern and southern regions.Different processing degrees also make influence on the contents of chemical compounds.The color parameter value can be used to evaluate the color of processed frankincense.

AIM: To investigate the affected proteins by tumor necrosis factor (TNF)-? in endothelial cells, and further explore the potential molecular mechanism of TNF-? on endothelial cells. METHODS: Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by a NO assay kit. Proteomic alterations were analyzed using two-dimensional electrophoresis, and peptide mass fingerprinting with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. RESULTS: NO production in HUVECs decreased significantly after TNF-? treatement. Proteomics analysis showed 21 protein spots were changed including 9 spots that were increased and 11 spots that were decreased after TNF-? stimulation, and 1 spot was only detected in TNF-? activated cell gels. CONCLUSIONS: The decreased expression of ecNOS by TNF-? might result in decrease in NO production. Up-regulated MAP/ERK kinase 3 expression might imply that TNF-? activates the expression of adhesion molecules. Cytoskeletal protein actin is also involved in TNF-? injuried HUVECs. Proteomic analysis can find some clues for identifying new potential target of TNF-?. [

OBJECTIVE: To establish a feasible evaluation index and method to identify composition of remaining metal particles on ferrochrome kitchen knife. METHODS: The small samples of remaining metal particles were rubbed from the knives using filter paper. The composition of remaining metal particles was detected by scanning electron microscopy and energy dispersive X-ray spectrometer (SEM-EDX) and GSR particle analysis function, using mathematical methods to calculate the ratio (relative amount) of Fe and Cr in remaining metal particles. RESULTS: The ratio (relative amount) of Fe and Cr of remaining metal particles had significant differences among most ferrochrome kitchen knives (P < 0.05). CONCLUSION Using GSR particle analysis function to quantitatively detect the ratio (relative amount) of Fe and Cr of remaining metal particles on ferrochrome kitchen knife, which can establish the feasible evaluation method to estimate such injury tool.
China ; Chromium/isolation & purification* ; Forensic Medicine/methods* ; Iron/isolation & purification* ; Metals/chemistry* ; Microscopy, Electron, Scanning/methods* ; Spectrometry, X-Ray Emission/methods* ; Wounds, Nonpenetrating/pathology*

OBJECTIVETo study the effects of Si-Wu-Tang on serum protein of blood deficient mice b y proteomicstechnique and study the enriching and regulating blood mechanism of Si-Wu-Tang on mocular level.

METHODThe blood deficient mice was induced by using a single dose of 3.5 Gy radiation from a 60Cogamma source, and high resolution two-dimensional polyacryamide gel electrophoresis (2-DE), computer-assisted image analysis, and mass spectrometry were used to detect regulated protein by Si-Wu-Tang.

RESULT12 lower and 4 higher protein in sera could be recovered by Si-Wu-Tang, 4 protein might be DNA-dependent protein kinase catalytic subunit, Dystrophin, KIF13A, dystonin. They play a part in DNA double-stranded break repair, recombination and modulation of transcription, transportation of mannose-6-phosphate receptor, etc.

CONCLUSIONSi-Wu-Tang can regulate serum protein in blood deficient mice, resulting in improving hematopoiesis and lessening irradiated injury.

Animals ; Autoantigens ; blood ; Carrier Proteins ; Cytoskeletal Proteins ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Dystonin ; Dystrophin ; blood ; Female ; Kinesin ; blood ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; Non-Fibrillar Collagens ; blood ; Plants, Medicinal ; chemistry ; Radiation Injuries, Experimental ; blood ; Radiation-Protective Agents ; pharmacology ; Whole-Body Irradiation

OBJECTIVETo observe the effects of siwu tang on protein expression of bone marrow of blood deficiency mice and provide the theoretical and experimental basis for understanding the molecular mechanism of blood enriching function of siwu tang.

METHODBlood deficiency mice were established by using 3.5 Gy 60Co gamma-ray. With proteomic technologies including two-dimensional electrophoresis, image analysis, in-gel digestion, peptide mass fingerprinting and bioinformatics the proteins of bone marrow of blood deficiency mice were isolated, analyzed, and identified.

RESULTSiwu tang could reverse 10 up-regulated and 4 down-regulated proteins of blood deficiency mice bone marrow. Seven of the proteins were likely to be lymphocyte specific protein 1, proteasome 26S ATPase subunit 4, hematopoietic cell protein-tyrosine phosphatase, glyceraldehyde-3-phosphate dehydrogenase, growth factor receptor binding protein 14 and lgals12 respectively.

CONCLUSIONSiwu tang can regulate the protein expression of bone marrow of blood deficiency mice and thus promote the growth and differentiation of hematopoietic cells and then exert its effects on blood enriching function.

Adaptor Proteins, Signal Transducing ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Phosphoproteins ; blood ; Proteins ; metabolism ; Proteome ; metabolism ; Proteomics ; methods ; Radiation Injuries, Experimental ; blood ; Whole-Body Irradiation

Arresten as a endogenous inhibitor of angiogenesis originated from the carboxyl-terminal 223 amino acids fragment of the non-collagen domain in alpha1 chain of human collagen IV. In order to get the soluble arresten with biological activity, the cDNA of arresten was cloned and expressed in Pichia pastoris. The produced arresten cDNA was amplified by PCR using primer P1:5'-AGGCCCCGATGGGTTGC-3', primer P2:5'-CTATAAG GCACTTTACGGTTTC-3'. The PCR products was cloned into pGEM-T vector, and sequenced. The arresten cDNA from pGEM-T vector was recombined with vector pPIC9 as pPIC9-arresten, used to transform E. coli DH5alpha, and the inserted arresten cDNA confirmed by agarose electrophoresis and sequencing. pPIC9-arresten was linearized by Sac I. Pichia pastoris GS115 was treated with PEG1000 (followed Invitrogen' s specification); transformed with linear recombined pPIC9-arresten. Pichia pastoris GS115 was culured on MD mediun, single clone was selected and the DNA from the single clone was extracted, used as template, characterized by PCR using the second pair primers P3:5'-CGCTCGAGAAAAGATCTGTTGATC-3', P4:5'-GCCCCGG ATCCTTATGTTCTFCTCATACAG-3'. The polynucleotides CTCGAGAAAAGA used as marker sequence was inserted into primer P3 for signal peptidase to cleave off the signal sequence correctively. The recombined Pichia pastoris GS115 was selected according to the results of PCR, cultured on MM and MD media and then in the BMGY media using methanol as inducer. Expressed arresten was analysed by SDS-PAGE. The soluble arresten expressed by Pichia pastoris gave apparent molecular weight in SDS-PAGE consistent with that calculated, and in matrigel gel it showed inhibitary activity on the tubulation of endothelial cell ECV-304 induced by tumor cell MDA-MB-435S. These results showesd arresten with biological activity is expressed successfully in Pichia pastoris GS115.
Arrestin ; genetics ; metabolism ; pharmacology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cells, Cultured ; Electrophoresis, Polyacrylamide Gel ; Endothelial Cells ; cytology ; drug effects ; Humans ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; metabolism ; pharmacology

OBJECTIVETo explore the role of inflammatory reaction mediated by p38-mitogen activated protein kinase (p38-MAPK) signal path on prevention and treatment of Parkinson disease (PD) model rats by electroacupuncture (EA).

METHODSThirty-two healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, eight rats in each one. The PD model was established in the model group and EA group by subcutaneous injection of rotenone in skin-back area (2 mg/kg, dissolved in sunflower oil, 2 mg/mL in density), while the injection of sunflower oil emulsion without rotenone at the same point and quantity as the model group was applied in the sham operation group. The normal group was not given any intervention. The EA treatment (continuous wave, 2 Hz in frequency, 1 mA in intensity, 20 min) was applied at "Fengfu" (GV 16) and "Taichong" (LR 3) in the EA group, once a day for continuously 14 days. No treatment was given in the other groups. The expression of tyrosine hydroxylase (TH), phosphorylated p38-MAPK, cyclooxygenase-2 (COX-2) in the substantia nigra were detected with immunohistochemical method.

RESULTSThere was typical PD ethology change in the model group. Compared with the normal group and sham operation group, the expression of TH positive neuron in the substantia nigra in the model group was significantly decreased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly increased (all P < 0.01). Compared with the model group, the expression of TH positive neuron in the EA group was apparently increased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly decreased (all P < 0.01).

CONCLUSIONThe EA therapy could obviously reduce the expression of inflammation mediator COX-2, inhibit the phosphorylation of p38-MAPK, reduce the damage of dopaminergic neurons in the rats with PD, and this effect may be related with the impact of p38-MAPK signal path

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Electroacupuncture ; Humans ; Male ; Parkinson Disease ; enzymology ; genetics ; therapy ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; enzymology ; Tyrosine 3-Monooxygenase ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism