Objective To detect the level of granulation tissue and expression of matrix metalloproteinase-1,3(MMP-1,3) in the tissues originated from both herniated and non-herniated intervertebral discs. Methods Forty specimens of herniated lumbar disc and twelve specimens of non-herniated one were stained with hematoxylin-eosin to determine the amount of granulation tissue, and immunohistochemically stained to detect MMP-1,3 expression. Results In specimens of sequestration and transligamentous extrusion, the amount of granulation tissue including macrophages,fibroblasts and capillary was more than that in the specimens of subligamentous extrusion and protrusion as well as non-herniated disc. Most of cells in granulation tissue and chondrocytes had MMP-1,3 expression. The expression level of MMP1,3 in sequestration and transligamentous extrusion specimens was significantly higher than that in subligamentous extrusion and protrusion, and in herniated specimens was also significantly higher than that in non-herniated specimens.There was no significant difference in the MMP1,3 expression level between the sequestration and transligamentous extrusion, between subligamentous extrusion and protrusion. Conclusion The expression of MMP1,3 was associated with the degradation and herniation of intervertebral disc. MMP1,3 may be involved in the pathogenesis of intervertebral disc degeneration and herniation.

Objective To investigate the expression and significance of vascular endothelial growth factor(VEGF) in the various types of degenerated lumbar discs and the normal control.Methods Forty-one cases of degenerated intervertebral disc tissue(the contained group with 17 cases and uncontained group with 24 cases) from surgical intervention for lumbar disc herniation and 13 cases of normal disc tissue from anterior approach for scoliosis and lumbar fracture were stained with hematoxylin and eosin.The expression of VEGF was tested by immunohistochemistry method.Results There was no capillary infiltration and VEGF positive expression in the control group.The VEGF positive rate was 41.2%(7/17) in the contained group,and 79.2%(19/24) in the uncontained group.In the contained group and uncontained group,the VEGF positive rate was higher than that in the control group(P

Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.

Objective To evaluate the therapeutic effects of autolngous bone marrow mesenchymal stem cells transplantation in the early phase of acute renal ischemia-reperfusion(I/R)injury in rabbits and their impact on tubular epithelial cell apoptosis.Method Bone marrow mesenchymal stem cells(MSCs)were isolated and cultured in vitro.Thirty rabbits were divided randomly into transplanted group,control group and sham-operated group,10 in each group.Rabbits of transplanted group were induced acute renal I/R injury by clamping bilateral renal arteries for 90 minutes,then transplanted MSCs labeled with BrdU immediately after the resumption of circulation in kidney.The rabbits of control group were induced acute renal I/R injury and then infused with saline instead.The sharn operated rabbits were transplanted autologous MSCs labeled with BrdU after sham operation.Fortyeight hours after transplantation,all rabbits were sacrificed.Renal functional and structural damage were evaluated.MSCs derived BrdU positive cells were determined by immunostalning.Tubular epithelial cell apoptosis was arialyzed by usign TUNEL.The SPSS version 11.3 Software was used for statistical analysis.ANOVA were used to ana lyze the data of renal function and tubular epithelial cell apoptesis.Data of renal structure darnage were analyzed by using the nonparametric Kruskal-Wallis test.Results Compared with control group,the renal functional and struc tural darnage were significantly ameliorated(24 h,BUN:F=7.483,Ser:F:15.091;48 h,BUN:F:17.741.Scr:F=61.865;P<0.05),tubular epithelial cell apoptods Wsa reduced significantly in transplanted group(F=135.495,P<0.01).MSCs derived BrdU positive cell were detected in the renal tissues of transplanted rabbits.Conelusiu Autologous bone marrow mesenchymal stern cells transplantation is curative for acute renal I/R injury in rabbits.Bone marrow MSCs might ameliorate renal I/R injury by reducing tubular epithelial cell apoptosis.

Objective To observe the release of inflammation-related factors after angiotensin Ⅱ (Ang Ⅱ ) stimulation in rat tubular epithelial cells (NRK-52E), to analyze whether these effects were mediated by TLR4-MyD88 pathway, and to reveal the novel mechanism of injury by Ang Ⅱ on NRK-52E cells. Methods After synchronization, cells incubated with AngⅡ (10-7 mmol/L) were used as the stimulation group, cells without stimulation were as normal control. To determine the role of TLR4 and the adaptor MyD88, equal number of NRK-52E cells was added with 10-5 mmol/L candesartan or 20 mg/L TLR4 blocking peptide for 1 h and then incubated with Ang Ⅱ (10-7 mmol/L) respectively. RT-PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression. Immunofluorescence and confocal microscopy were used to observe TLR4 protein expression. ELISA was used to detect the concentration of tumor necrosis factor-alpha (TNF-α) and heat shock protein 47(HSP47) in cell supernatant respectively. Results TLR4 and MyD88 were highly expressed in Ang Ⅱ-induced NRK-52E cells (P＜0.01), and the TNF-α and HSP47 levels were also increased markedly compared with control group (P＜0.01). In NRK-52E cells that were pre-incubated with candesartan, TLR4 and MyD88 expression were obviously inhibited,subsequently, HSP47 and TNF-α production decreased remarkably compared with Ang Ⅱ group (P＜0.01). TLR4 blocking peptide had the similar effect in a dose-dependent manner, in which its effect was dependent on inhibiting TLR4-MyD88 expression. Conclusion The mechanism of Ang Ⅱ -induced injury effect on NRK-52E cells is related to the increase of TLR4-MyD88 activity,which is followed by the enhance of TNF-α and HSP47 expression. This process is inhibited by candesartan via modulation of innate immune pathway.

BACKGROUND:Spine infection models are rarely reported in the literature. There are fewer reports on the imaging and histological observations of the animal model of spinal infection.
OBJECTIVE:To develop a canine model of pyogenic spinal infection.
METHODS:Twelve Chinese dogs were adopted in the study. The dogs were in the right arm recumbent. The surgery was conducted by retroperitoneal approach to expose T12/L1 intervertebral space. The intervertebral discs that exposed the intervertebral space were partial y removed, and the end plate of the adjacent two vertebrae was deliberately curetted out. Suspension with different concentrations of Staphylococcus aureus and sodium morrhuate were injected into the dog intervertebral space. The aim of removal of partial intervertebral discs and destruction of endplate was to make the vertebral body bone better in contact with the bacteria. The same operation was conducted in L5/6 clearance of the animal spine. At 14 days after surgery, animals were sacrificed. The infected intervertebral disc and adjacent vertebral bodies were taken out. Bacterial contents of specimens were quantitatively determined. During sacrificing, partial liver tissues and blood samples were col ected for bacterial culture. The occurrence of systemic infection was observed.
RESULTS AND CONCLUSION:(1) Spondylodiscitis of the lumbar spinal column was consistently produced in 90%(9/10) the sites chal enged with 102 colony-forming units (CFU) Staphylococcus aureus. Liver biopsy and blood culture did not show any signs of systematic infections. (2) Two out of four animals implanted the suspension with a concentration higher than 103 CFUs died within 3 days post-implantation. (3) These results suggested that a new canine model of pyogenic spinal infection was developed. This model could be used to test the efficacy of different anti-infection strategies.

Objective To summarize the surgical procedures of unroofed coronary sinus syndrome with endocardial cushion defect and persistent left superior vena cava.Methods From 2000 to 2008,753 cases of endocardial cushion defect underwent surgical repair.Among them 15 cages(2.0%)were associated with persistent left superior vena cava and unroofed coronary sinus syndrome.The drainage of the persistent left superior vena cava and coronary vein were corrected simultaneously during the operation by reestablishing the roof of the coronary sinus.The persistent left superior vena cava were drained to right atrium by intra-atrial tunnel with pericardial patch or left atrium folding in 14 cases and it was ligated in the remain one.Most of the coronary veins were drained into right atrium after the correction of cardiac defect.Results One patient died of severe pulmonary infection early after operation.Echocardiogram indicated that the velocity of left superior vena cava increased in the site of the roof of left atrium in one patient after operation.No obstruction of vena cava and pulmonary vein and remnant shunt between the atriums were found in the other 14 cases.There were no death and other complications during3 months to 6 years of follow-up.Conclusion The surgical procedures of unroofed coronary sinus syndrome with endocardial cushion defect and persistent left superior vena cava is difficult and good result can be expected by reestablishing the roof of the coronary sinus.

OBJECTIVETo develop a convenient, practical low-cost and efficient Ganoderma spore collector.

METHODThe spore collector was made from common materials such as white cardboard and oil-lustrous paper, temperature and humidity were used as indexes to study the effect of the collector on the growth environment of Ganoderma and spore collection.

RESULTThe spore collector developed could effectively separate Ganoderma fruit bodies from the outside to form an independent closed space and stop free flow of spores. The use of the collector had few effects on temperature and humidity that influenced the growth of G. spp. and development of the fruit bodies. In addition, the fluctuation of the relative humidity inside the collector tended to be small.

CONCLUSIONThis collector could efficiently collect quality spores and the yield of spores accounted for 38.3% of the total yield of spores and fruit bodies when this collector was applied on a large scale.

Agriculture ; methods ; Equipment Design ; Equipment and Supplies ; Fruiting Bodies, Fungal ; growth & development ; Ganoderma ; growth & development ; Plants, Medicinal ; growth & development ; Spores, Fungal ; growth & development

Objective This study is to compare flap-viability-related complications, coverage reach, recon-struction outcomes and donor-mobidities between distally-based peroneal artery perforator-plus fasciocutaneous (DPAPF)flap and distally-based posterior tibial artery perforator-plus fasciocutaneous(DPTAPF)flap for recon-struction of soft-tissue defects over the distal lower leg, ankle and foot, and thus provide evidence for selection of the flaps. Methods Between April, 2002 and February, 2012, 216 and 59 patients underwent the reconstructions with DPAPF flaps(peroneal group)and DPTAPF flaps(posterior tibial group)respectively. We subdivided the distal lower leg, ankle and foot into 12 subregions. In all the patients, flap-viability-related complications and its potential risk factors(including age,sex,etiology,location of top edge,location of pivot point,length and width of both the skin is-land and adipofascial pedicle, length-width ratio, and total length), coverage reach(the subregion in which the most distal part of the reconstructed defect lies),duration of flap elevation and hospital stay were compared between the two groups. In patients with at least 3 months postoperative follow-up, comparative study of reconstruction outcomes, pa-tient's satisfaction with flap appearance and donor-site morbidities were performed between the groups. Results Partial necrosis rate in the peroneal of the posterior tibial group were 12.0 percent versus 20.3 percent,respectively(P> 0.05). Marginal necrosis and overall complication (including partial and marginal necrosis)rates in the peroneal group(1.9 percent and 13.9 percent, respectively)were significantly lower than those in the posterior tibial group (8.5 percent and 28.8 percent,respectively)(P<0.05).Incidence of partial necrosis of the flaps for the defects over subregions 7 to 10 in the peroneal group(7 of 41)was significantly lower than that in the posterior tibial group(2 of 2).There was no difference in reconstruction outcomes and patient's satisfaction with flap appearance in both groups(P >0.05).Incidences of hypertrophic scar,itching and pigmentation at the donor site were significantly lower in the peroneal group(P<0.05). Conclusion DPAPF flap is superior to DPTAPF flap in reliability,safe coverage reach and less donor-site morbidities.The former is recommended as the first choice when local pedicle flaps are considered to recon-struct soft-tissue defects over the distal lower leg,ankle and foot.

Objective To compare differences of SRS-22 scores between male and female AIS patients.Methods From October 2007 to April 2009,298 AIS patients from 7 spine centers completed simplified Chinese edition of SRS-22 questionnaire,including 62 males and 236 females with an average age of 15.7 years old,and the average Cobb angle was 52.8°.Of 62 male patients,34 were from urban areas and 28 rural areas.Among 236 female patients,132 were from urban areas and 104 rural areas.A comparison was conducted in terms of age,Cobb angle,the ratio of urban population to rural population,and the scores of all domains and the subtotal scores of SRS-22 questionnaire between male and female patients.Results There was no difference of age,Cobb angle and the ratio of urban population to rural population between male and female patients(P＞0.05).The scores of four domains and subtotal scores of SRS-22 questionnaire were significantly higher in male patients when compared with female patients(P＜0.05).Conclusion Gender difference should be taken into the consideration when estimating the quality of life in AIS patients by SRS-22 questionnaire.The quality of life in male patients with AIS was better than that of female patients.