AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

Objective To discuss the short-term currative effect of horizontal osteotomy across injured vertebral forⅤdegree old frac-ture-dislocations of thoracolumbar spine.Methods The data of 4 patients who received horizontal osteotomy across injured vertebral internal fixation in our hospital from April 2010 to October 2012 were retrospectively analyzed,of which 1 case with T8-9 fracture dislocation,2 cases with T10-11 fracture dislocation,1 case with T11-12 fracture dislocation.The Franke1 classification of all patients were grade A.Results All 4 cases were achieved completely fracture reduction and followed-up for 2-14 months(an average of 7.4 months),who could get sitting posi-tion with the support of brace 1 week after operation.The postoperative follow-up indicated that all 4 cases got fusion,but there was no improve-ment for Franke1 classification.Conclusion The method of horizontal osteotomy across injured vertebral forⅤdegree old fracture-dislocations of thoracolumbar spine has good short-term clinical effect,with the advantages of shorter operation time,less bleeding and good reduction effect.

BACKGROUND:The viability of human umbilical cord-derived mesenchymal stem cel s is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cel ular transplantation. However, reasons for this are stil unknown. OBJECTIVE:To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cel s viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS:Human umbilical cord-derived mesenchymal stem cel s were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracel ular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracel ular oxidative stress levels after storage. Cel adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION:The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cel s were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were al reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cel s viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cel s is one of the reasons for reduced cel viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cel s.

Objective To study the effect of different rod curvature on the postoperative stability and stress of thoracolumbar junction fracture using the finite element simulation.Methods (1) Thoracolumbar finite element model of T11 to L1 from three-dimensional CT data of a 30-year-old healthy male volunteer was established,including the assignment of cortical bone,cancellous bone,disc,ligaments and facet joints.On this basis,the T12fracture model was also established.T11 to L1 bilateral pedicle screw fixation was loaded,and the rod connection was divided into straight rod group and pre-bended rod group (lordotic 15°-25°) according to angle of the rod.A 400 N stress was loaded on the top of the upper endplate of T11 to simulate the upper part body gravity,while applying a 10 N · m torque to generate flexion and extension,lateral flexion and rotation.Stress distribution of different methods of pre-bending for thoracolumbar fractures after reduction was compared.(2) A retrospective cohort analysis was made on 56 cases of thoracolumbar fractures surgically treated from July 2012 to July 2016,including 31 cases in straight rod group and 25 cases in pre-bended rod group.Two groups were compared in angle between adjacent level before operation,after operation and at final follow-up.Results (1) In flexion,extension,lateral bending and rotation,both rod bending methods effectively controlled the thoracolumbar junction displacement.The peak stress of connecting rod (151,315,369,377 MPa respectively) in pre-bended rod group was lower than that in straight rod group (110,239,281,189 MPa respectively) (P ＜ 0.05),and straight rod group appeared relatively obvious stress concentration.(2) Mean follow-up time was 21.4 months (range,4-33 months).Preoperative kyphosis angle was (21.7 ± 7.4)°in straight rod group and (20.3 ± 6.8)° in pre-bended rod group (P ＞ 0.05).Postoperative lordosis angle in straight rod group was (3.3 ± 1.2) °versus (8.3 ± 2.8) ° in pre-bended rod group (P ＜ 0.05).At the final follow-up,the lordosis angle in straight rod group was reduced by (8.7 ± 2.3) ° versus (3.9 ± 1.7)°in pre-bended rod group (P ＜0.05).Implant failure was seen in 3 cases in straight rod group,but there was no implant failure in pre-bended rod group.Conclusion Pre-bended (lordotic 15°-25°) and pre-loaded rods used in internal fixation of thoracolumbar fractures may reduce the stress of rods,decrease the incidence of implant failure and facilitate the recovery of spine stability.

Objective To compare the feasibility and efficacy of intervertebral wedge osteotomy and pedicle subtraction osteotomy (PSO),vertebral column resection (VCR),Smith-Petersen osteotomy (SPO) for the treatment of severe kyphosis and scoliosis.Methods The data of 38 cases of severe kyphosis and kyphoscoliosis were retrospectively analyzed from January 2010 to February 2016,including 22 males and 16 females.According to the osteotomy mode,PSO,SPO,VCR and intervertebral disc wedge osteotomy were used to collect the average number of fixed phases,volume of bleeding,length of stay,length of hospital stay,improvement of main cobb angle,improvement of ODI score,and Frankel classification to evaluate the efficacy.Results There were no significant differences in the overall operative time between the four groups.The average number of fixation in 18 patients with SPO was (9.4±3.9) segments,the blood loss was (3 000±410) ml,the average Cobb angle was improved by 55.3％± 9.5％,the average postoperative hospitalization was (14.6±4.9) days,the improvement rate of ODI was 42.1％±7.4％,all the patients were improved to Frankel E;The average number of fixation in 5 patients with PSO was (7.6± 1.5) segments,the blood loss was (4 360± 1 161) ml,the average Cobb angle was improved by 58.9％ ± 15.1％,the average postoperative hospitalization was (18.2±7.0) days,the improvement rate of ODI was 41.3％±9.6％.One Frankel C patient was improved to Frankel D,others remained to be Frankel E as the same as pre-operation;The average number of fixation in 4 patients with VCR was (6.2±2.6) segments,the blood loss was (3 750+ 1 848) ml,the average Cobb angle was improved by 83.9％± 10.9％,the average postoperative hospitalization was (21±7.2) days,the improvement rate of ODI was 39.6％± 18.1％.Three Frankel D patients were improved to Frankel E and one Frankel C patient was improved to Frankel D;The average number of fixation in 11 patients with IWO was (7.1 ± 2.7) segments,the blood loss was (2855±1046) ml,the average Cobb angle was improved by 59.6％±22.05％,the average postoperative hospitalization was (13.5±2.7) days,the improvement rate of ODI was 51.3％±8.3％.One Frankel C patient was improved to Frankel D,eight Frankel D patients were improved to Frankel E,other patients remained to be Frankel E;The mean follow-up time was 25.2 months in 11 patients underwent intervertebral wedge osteotomy.All the patients had successful spinal fusion and no failure of internal fixation.Conclusion Intervertebral wedge osteotomy for the treatment of scoliosis and kyphosis could reduce surgical injury to obtain good biomechanics and surgical result.

To study the protective effect of Danqi Piantan capsule ( DPC) and its antelope horn substitution (DPCAS) on the cerebral ischemia, in order to preliminary study the possibility of replacing antelope horn with artificial bezoar. In this study, the left middle cerebral artery occlusion (MCAO) was adopted. Totally 150 SD rats were randomly divided into 5 groups: the sham operation group, the model group, the Danqi Piantan capsule (DPC) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn (DPCRA) group (0.246 g x kg(-1) x d(-1)), the Danqi Piantan capsule without antelope horn and with double artificial bezoar (DPCDB) group (0.246 g x kg(-1) x d(-1)). The MCAO model was prepared 1 h later after the administration on the 5th day. At 24 h after the operation, the inner canthus blood was collected to determine the serum superoxide dismutase (SOD) activity and the endothelin (ET) content. At 72 h after the operation, the cerebral infarct size and the cerebral index were determined by TTC-staining. The fluorescent quantitative PCR method was used to detect brain Bcl-2, Caspase-3, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions. The mmunohistochemical method was used to detect ICAM-1, IL-1β, TNF-α, IL-6 expressions in ischemic penumbra. According to the results, compared with the model group, DPCDB and DPC groups showed almost consistent results, indicating both of the two group can significantly improved cerebral infarction index and cerebral index (P < 0.05), increase the serum SOD activity (P < 0.05), decrease the serum ET level and Caspase-3 expression, IL-1β, P-selectin, E-selectin, ICAM-1 mRNA expressions in brain tissues (P < 0.05) and expressions of ICAM-1, IL-1,6, TNF-α, IL-6 positive cells in ischemic penumbra (P < 0.05) and increase the Bcl-2 expression (P < 0.05). The DPCRA group showed much lower impacts on indexes than DPCDB and DPC groups. This suggests that DPCDB and DPC reveal similar efficacies and antelope horn in Danqi Piantan capsule can be substitutes by artificial bezoar.
Animals ; Antelopes ; Bile ; chemistry ; Biological Factors ; administration & dosage ; chemical synthesis ; chemistry ; Brain ; drug effects ; metabolism ; Caspase 3 ; genetics ; metabolism ; Drug Compounding ; Horns ; chemistry ; Humans ; Infarction, Middle Cerebral Artery ; drug therapy ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Male ; Medicine, Chinese Traditional ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; genetics ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Objective: To investigate the effects of acupuncture on constipation in rats. Method:Compound diphenoxylate was used to set up rat model of constipation. After needling Heguand Housanli, the time to first bowel movement, and stool grains quantity and weight in 12 rats were observed. Results: The time to first bowel movement was delayed, and the stool grains quantity and weight were reduced in the model rats. Acupuncture could significantly shorten the time to first bowel movement, and increase stool grains quantity and weight.Conclusion: Acupuncture can improve the defecating abilities in rats model of constipation.

The in vitro cell culture experiment was conducted to study the effect of Danqi Piantan capsule (DPC) and DPC dislodge the antelope horn with artificial bezoar double (DPCBD) on nerve regeneration and blood vessel regeneration and preliminarily investigate the possibility of substituting antelope horn in DPC with artificial bezoar. In this experiment, rats were randomly divided into 5 groups: the blank serum control group, the model group, DPC groups (0.306 g x kg(-1) x d(-1), the same below), DPC remove of antelope horn (DPCRA) groups and DPCBD groups. Brain microvascular endothelial cells cultured in vitro (BMEC), astrocytes and neural stem cells (NSC) were co-cultured to simulate neurovascular unit, label neurons with microtubule associated protein III (β-tubulin III) antibody and lable astrocytes with glial fibrillary acidicprotein (GFAP). ELISA was used for the detection of the content of BMEC lactate dehydrogenase instrument method (LDH), the inverted phase contrastmicroscope was adopted to observe the formation of BMEC tube like structure, the number of leukocytes and leukocytes adherent to BMEC were counted under the microscope, the expression levels of β-tubulin III and the ratio of GFAP positive cells was detected with inimmunofluorescence, and RT-PCR method was used to detect NGF, BDNF, VEGF and VEGFr-2 mRNA. According to the result, compared with the model group, both DPC and DPCBD can reduce LDH leakage, promote the formation of BMEC tube like structure, inhibit leukocytes and their adhesion to BMEC, increase the β-tubulin III positive cell differentiation proportion (P < 0. 01), reduce the proportion of GFAP positive cells (P < 0.01), increase the expressions of co-cultured NGF, VEGF, BDNF and VEGFr-2 mRNA to a certain extent, with the most significant difference on NGF and VEGF mRNA expressions (P < 0.05) and the same efficacy in both groups. DPCRA groups showed less impact on all indexes than that of DPCBD and DPC groups. The same efficacy of DPCBD and DPC on nerve regeneration and angiogenesis suggested that antelope horn in DPC can be substituted by artificial bezoar.
Animals ; Antelopes ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Cattle ; Cells, Cultured ; Drug Substitution ; Endothelial Cells ; drug effects ; metabolism ; Gallstones ; chemistry ; Horns ; chemistry ; Male ; Medicine, Chinese Traditional ; Nerve Growth Factor ; genetics ; metabolism ; Neural Stem Cells ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

Aged ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Hemangioendothelioma, Epithelioid ; pathology ; Humans

BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.