Basic fibroblast growth factor (bFGF) is a multi-potential growth factor whose biological activities depend on its receptor's intrinsic tyrosine kinase activity and second messengers such as the mitogen activated protein kinases. Heparin sulfate proteoglycans (HSPGs) have been demonstrated to enhance or inhibit bFGF activity. The response elicited by HSPG is related to the relative concentrations and binding kinetics for bFGF of the various pools of HSPG. The type of cellular response might depend on the specific HSPG and FGF receptor expressed on the cell surface. The specific core protein of HSPG, and tissue specific differences in heparin sulfate modification result in altered bFGF regulation.

Cigarette smoke extract (CSE) contains abundant oxidants and free radicals. Oxidative stress caused by cigarette smoking results in the destruction of the alveolar cell walls and emphysema. However, there exists discrepancy about how CSE works in the process. In the present study, we observed the effect of CSE on the cell growth of type II alveolar epithelial cell-derived A549 cell line, and provided molecular understanding of this effect. The MTT assay results showed that CSE decreased the cell viability of A549 cells in a dose- and time-dependent manner, and cell cycle was arrested in G(1)/S phase. Furthermore, CSE-induced apoptosis of A549 cells was verified by Hoechst 33258 staining, electron microscopy in morphology, and the appearance of DNA fragmentation and annexin V-FITC/propidium iodide (PI) staining assay at molecular level. It was found that CSE treatment resulted in the upregulation of Fas/APO-1 receptor and activation of caspase-3. CSE also initiated accumulation of intracellular reactive oxygen species, which was detected by laser confocal microscopy. Taken together, CSE could inhibit the cell growth and induce apoptosis of A549 cells through Fas receptor pathway. Oxidative stress caused by CSE may be the radical factor leading to apoptosis as well as cell growth inhibition in alveolar epithelial cells.
Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Epithelial Cells ; drug effects ; Humans ; Lung Neoplasms ; pathology ; Pulmonary Alveoli ; cytology ; drug effects ; pathology ; Smoke ; adverse effects ; Tobacco ; toxicity

The technique conditions of decolonization of fermentation broth were successively optimized using single factor assay and orthogonal layout method in Cordyceps jiangxiensis. The optimal condition of decolorization was investigated to be 3g/100mL active carbon, 5 min absorption time, pH5 of fermented broth and 25℃absorption temperature. Under the optimal condition, the maximum decolorization rate of fermented broth reached 89. 6% , simultaneously 10. 7% consuming rate of exopolysaccahride was minimum. Subsequently, the extract condition of exopolysaccharide of C. jiangxiensis was further optimized by orthogonal layout design. The maximum exopolysaccharide production was 0. 38 g/L under the optimal condition, i. e. firstly fermented filtrate decolorized and deproteined was concentrated to 1/7 of its total volume, secondly concentration broth was mixed with four times its volume of absolute ethanol and stirred vigorously, lastly precipitation of exopolysaccharide proceeded at 4℃for 16 hrs and the exopolysaccharide collected by centrifugal ion and dryness.

OBJECTIVETo construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells.

METHODSThe designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method.

RESULTSThe proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation.

CONCLUSIONSThese new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types.

Antigens, Surface ; analysis ; Base Sequence ; Biological Assay ; Cells, Cultured ; DNA ; analysis ; genetics ; Gene Expression Profiling ; methods ; Humans ; Lipopolysaccharide Receptors ; analysis ; Lymphocyte Antigen 96 ; Membrane Glycoproteins ; analysis ; Molecular Probe Techniques ; Molecular Sequence Data ; Monocytes ; metabolism ; RNA Probes ; analysis ; genetics ; Receptors, Cell Surface ; analysis ; Receptors, Immunologic ; analysis ; Ribonucleases ; Toll-Like Receptor 4 ; Toll-Like Receptors

OBJECTIVETo investigate the effects of intrathecal ouabain and tizanidine injection for treatment of neuropathic pain in rats.

METHODSMale SD rats weighing 250-300 g were randomly divided into 5 groups (n = 6), namely the control group, ouabain group, tizanidine group, combined ouabain and tizanidine injection group, and the antagonist group. Intrathecal catheter was implanted 7 days before spinal nerve ligation to establish the neuropathic pain model. Mechanical withdrawal threshold (MWT) before and after intrathecal administration of the agents was recorded in the rats. Isobolographic analysis was performed to evaluate the interactions between the agents.

RESULTSIntrathecal injection of ouabain (0.25-5 microg) or tizanidine (0.5-5 microg) alone produced dose-dependent analgesic effect against the neuropathic pain (P < 0.05). Isobolographic analysis revealed a synergistic interaction between ouabain and tizanidine. Intrathecal pretreatment with atropine (5 microg) or yohimbine (20 microg) antagonized the effects of ouabain and tizanidine administered alone or in combination (P < 0.05).

CONCLUSIONIntathecal injection of ouabain or tizanidine produces dose-dependent analgesic effects against neuropathic pain, and their synergistic effect after combined injection probably involves the cholinergic transmission and alpha2 receptor.

Analgesics ; administration & dosage ; Animals ; Clonidine ; administration & dosage ; analogs & derivatives ; Injections, Spinal ; Ouabain ; administration & dosage ; Pain ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Nerves ; injuries

OBJECTIVETo select the optimal pregnancy time window of embryonic pig skin precursor tissue for xenotransplantation and study its ability in wound repair.

METHODSSkin precursor tissues were obtained from pig fetus of fetal age of 35, 42, 56, 70 days, and were minced into microskin and transplanted to dorsal wounds of BALB/c nude mice, then they were covered with residual skin after plastic surgery of patients or adult pig skin (white). The characteristics of growth and development were observed after transplantation. Pathological examination was performed on 6 and 12 post operation weeks respectively to observe the tissue structure and tumorigenicity.

RESULTSSkin precursor tissues from fetal pig survived and developed after transplantation, and the microskin fused. New tissue area from skin precursor tissues with fetal age of 42 days was (47 +/- 6) mm2, which was higher than that of 35 days (18 +/- 8 mm2), 56 days (31 +/- 12 mm2), 70 days (20 +/- 8 mm2, P < 0.05). The skin precursor developed into "intact skin" with hair, sebaceous glands and sweat glands, and melanocytes were also detected in epidermis. The newly-grown skin tissue included epidermal and dermal layer, and obvious dermal papillae. Teratoma was not found after transplantation in skin precursor tissue with fetal age of 56, 70 days.

CONCLUSIONFetal pig skin precursor tissue with fetal age of 56 days can be used to repair wound as xenotransplantation.

Animals ; Fetal Tissue Transplantation ; Fetus ; Gestational Age ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Skin Transplantation ; Swine ; Transplantation, Heterologous ; Wound Healing

BACKGROUNDThe traditional lamina osteotomy replantation method is prone to nerve root injury and low back pain recurrence. Our team has proposed a modified approach that improves the osteotomy site and its fixation procedure. The aim of this study was to evaluate the clinical efficacy of traditional and modified lamina replantation methods in treating unstable lumbar disc herniation.

METHODSFrom March 2008 to August 2011, 124 patients with unstable lumbar disc herniation were enrolled and randomly divided into the following two groups according to random digital table: group A (traditional group) consisting of 61 patients who underwent traditional laminectomy replantation, and group B (modified group) consisting of 63 patients who underwent modified lamina replantation. Both surgeries were performed by the same surgeons. The two groups had no significant difference in gender, age, symptoms, time of onset and the prominent segment. Visual analogue scale (VAS), Oswertry disability index (ODI), and Japanese Orthopaedic Association (JOA) scores, operative time, blood loss, complication rate, radiographic healing rates, and low back pain recurrence rates were compared between the two groups.

RESULTSThere were 121 patients followed up for more than one year, and the follow-up rate was 97.6%. Nerve injury occurred in two patients (3.3%) in the modified group and 12 patients (20.0%) in the traditional group. Dural injury occurred in one patient (1.6%) in the modified group and seven patients (11.7%) in the traditional group. Pseudarthrosis occurred in two patients in the modified group and in 18 patients in the traditional group with 1-year fusion rates of 96.7% and 70.0%, respectively. Recurrence of lower back pain after one year was noted in three patients (4.9%) in the modified group, and in 15 (25.0%) in the traditional group. Leg pain recurrence was noted in one patient (1.6%) in the modified group and in three cases (5.0%) in the traditional group. The one-year healing rates of nerve injury, dural injury, replantation lamina and low back pain recurrence rates after one year were significantly different (P < 0.05) between the two groups. At two weeks, three months, six months and one year postoperatively, both groups had significant improvement in VAS, ODI, and JOA scores from their preoperative values (P < 0.05). No significant difference was detected between the short term postoperative scores between groups A and B (P > 0.05). However, a significant difference was found one year later (P < 0.05).

CONCLUSIONSCompared to the traditional approach, the modified technique for lamina replantation showed lower rates of dural and nerve damage, a higher lamina healing rate, a lower back pain recurrence rate, and better clinical scores. It is a safe and effective operation for lumbar spine surgery.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; surgery ; Laminectomy ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Osteotomy ; methods ; Replantation ; methods

Objective To identify the optimal mono-energetic enhanced spectral CT for renal cortex in cortical phase based on the iodine concentration. Methods Fifty patients with normal renal function received the abdominal enhanced spectral CT examination.The iodine concentration and CT values of the multiple mono-energetic spectral images were measured on renal cortex in cortical phase,and the correlation between the iodine concentration and the CT values and the coefficient of variation(CV)were analyzed. Results The correlation analysis demonstrated that the correlation coefficient was 0.994,0.994,0.993,0.987,0.976,0.960,and 0.938 between mono-energetic spectral CT images(40-100 keV with interval 10 keV,respectively)and iodine concentration(all
Contrast Media ; Humans ; Iodine ; Kidney Cortex/diagnostic imaging* ; Radiographic Image Interpretation, Computer-Assisted ; Tomography, X-Ray Computed

Objective To investigate the intra-and inter-observer reproducibility of iodine concentrations of abdominal parenchymal organs based on spectral CT.Methods The water-free iodine images of the venous phase were retrospectively obtained from 50 patients with abdominal dynamic spectral CT scans.The iodine concentrations were measured in the left,right and caudate lobes of liver,spleen,pancreas and bilateral kidneys.Intraclass correlation coefficient(ICC)and Bland-Altman plot were employed to analyze the intra-and inter-observer reproducibility.Results The intra-observer ICCs of the left,right and caudate lobes of liver,spleen,pancreas,and left and right kidneys were 0.938(0.894,0.965),0.932(0.884,0.961),0.939(0.895,0.965),0.947(0.909,0.970),0.912(0.851,0.949),0.946(0.906,0.969)and 0.907(0.842,0.946),which indicated good intra-observer reproducibility.The inter-observer ICCs of the left,right and caudate lobes of liver,spleen,pancreas,and left and right kidneys were 0.947(0.909,0.970),0.927(0.875,0.958),0.943(0.902,0.968),0.956(0.924,0.975),0.934(0.887,0.962),0.927(0.875,0.958)and 0.892(0.818,0.937),which indicated good inter-observer reproducibility.Bland-Altman plots presented that more than 95% points of the intra-observer differences located within 95% CI of limits of agreement for the caudate lobe of liver,spleen,pancreas and bilateral kidneys,which was same as inter-observer differences of the caudate lobe of liver,spleen and right kidney.Conclusion The iodine concentration measurement based on the spectral CT presented good intra-and inter-observer reproducibility for the caudate lobe of liver and spleen.
Humans ; Iodine ; Observer Variation ; Reproducibility of Results ; Retrospective Studies ; Tomography, X-Ray Computed

Objective To evaluate the optic nerve impairment using MRI histogram texture analysis in the patients with optic neuritis.Methods The study included 60 patients with optic neuritis and 20 normal controls. The coronal T2 weighted imaging (T2WI) with fat saturation and enhanced T1 weighted imaging (T1WI) were performed to evaluate the optic nerve. MRI histogram texture features of the involved optic nerve were measured on the corresponding coronal T2WI images. The normal optic nerve (NON) was measured in the posterior 1/3 parts of the optic nerve. Kruskal-Wallis one-way ANOVA was used to compare the difference of texture features and receiver operating characteristic (ROC) curve were performed to evaluate the diagnostic value of texture features for the optic nerve impairment among the affected optic nerve with enhancement (ONwEN), affected optic nerve without enhancement (ONwoEN), contralateral normal appearing optic nerve (NAON) and NON.Results The histogram texture Energy and Entropy presented significant differences for ONwEN vs. ONwoEN (both P=0.000), ONwEN vs. NON (both P=0.000) and NAON vs. NON (both P<0.05). ROC analysis demonstrated that the area under the curve (AUC) of histogram texture Energy were 0.758, 0.795 and 0.701 for ONwEN vs. ONwoEN, ONwEN vs. NON and NAON vs. NON, AUC of Entropy were 0.758, 0.795 and 0.707 for ONwEN vs. ONwoEN, ONwEN vs. NON and NAON vs. NON.Conclusions The altered MRI histogram texture Energy and Entropy could be considered as a surrogate for MRI enhancement to evaluate the involved optic nerve and normal-appearing optic nerve in optic neuritis.
Adolescent ; Adult ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Optic Nerve ; diagnostic imaging ; Optic Neuritis ; diagnostic imaging