The authors adopted an auto control experimental research on 18 male adult dogs with 36 sides of femoral heads and necks which were divided into two groups. They were separately made models of old fracture of femoral neck and avascular necrosis of famoral head. Two wecks later, The free fibula with microvascular an astomosis was tranplanted. All experimental dogs were examined with X-ray, pathohitology, electron microscopy, 99~mTc-MDP scanning and tetracycling labeling after operation.The experiment showed that the free fibular graft with mierovascular anastomosis could provided a new system of vascula supply to the injured femoral head. The hard free fibula and the metallic internal fixation were inserted in femoral head and neck. After the Operation, in the fracture area the fibula formed a bone-bridge which not only diminished the stress force but also prevented rotation displacement of the bone fracture The periosteum of the transplanted fibula also took part in the repair course of avascular nerosis of femoral head.

118 cases with uper and median partialfibulectomy were reported in this paper. Of them50 cases were followed up 1～8 years. The clinicalfunction, foot arch and X-ray of bilaleral ankleswere examined. Ankle joint function was mea-sured with the instrument made by ourself. Afteranalysis and valuation of the results, our conclu-sion is that uper and median partial fibulectormydoes not affect the ankle joint function. Thereforethe fibula being considered as a donor bone is ac-ceptable.

Objective To observe the effects of estrogen on behavior and 5-HT in periaqueductal gray (PAG)in migraine model rats. Methods Tewnty-four ovariectomized Wistar rats were divided randomly into four groups:control group (Group A),migraine group(Group B),low dose estradiol-treated ovariectomized group(Group C),high dose etradiol-treated ovariectomized group(Group D).After 1 week,the rats in Group B,C and D were injected with nitroglycerine 10 mg?kg-1 subcutaneously to make migraine rat models,the rats in Group A were given peanut oil alike,and the behavior changes were observed.2 h after injection,the rats were killed and the midbrains were separated and then 5-HT immunohistochemical staining was performed.Results Behavior: compared with Group B,the degrees of red-calws,red-ears and red-tail rats in Group D relieved obviously,the times of climbing hutch and scratching head were much fewer,while the rats in group C showed no significant difference;Immunohistochemical staining:compared with Group A,the 5-HT-positive neurons expression in PAG of Group B and C were more obviously(P

Objective:To study the effect of 3-n-butylphthalide (NBP) on the expression of brain-derived neurotrophic factor (BDNF) in the hippocampal CA1 region of the vascular dementia (VD) rats,and to explore its protective effect on VD.Methods:Eighty healthy Wistar rats were equally and randomly assigned to sham operation group,NBP control group (sham operation + NBP injection),VD group (VD models),NBP treatment group (VD models + NBP injection) (n=20).Each group was divided into four subgroups (n =5):1,2,4,and 8 weeks after operation groups.The VD rat models were established by using permanent bilateral common carotid artery ligation.After consciousness,the rats in NBP treatment group and NBP control group were intraperitoneally injected with 5 mg · kg-1 · d-1 NBP for consecutive 7 d.The rats in VD and sham operation groups were intraperitoneally injected with 0.2 mL · d-1 saline for consecutive 7 d.At 1,2,4,and 8 weeks after operation,the rats in each group were decapitated.The brains were obtained,and then the hippoeampus tissues were isolated.The BDNF expression levels in the hippocampal CA1 region were determined using real-time quantitative PCR and immunohistochemistry methods.Results:At 2,4,and 8 weeks after s operation,the expression levels of BDNF mRNA in the hippocampus of the rats in VD group were significantly higher those in sham operation group (P＜ 0.05);at 4 and 8 weeks after operation,the expression levels of BDNF mRNA in the hippocampus of the rats in NBP treatment groups were significantly higher than that in VD group (P＜ 0.05).The immunohistochemistry results showed that the expression level of BDNF protein in the hippocampal CA1 region of the rats in VD group was higher than that in sham operation group at 4 weeks after operation (P＜ 0.05);the expression level of BDNF protein in the hippocampal CA1 region of the rats in NBP treatment group was higher than that in VD group at 8 weeks after operation (P＜0.05).Conclusion:The BDNF expression is increased in the hippocampal CA1 region of the rats with VD after the neurons were injured by ischemia.NBP can increase the BDNF expression level in the hippocampal CA1 region of the VD rats and protect the nerves.

OBJECTIVETo study the inhibitory effects of insulin on nuclear factor-kappa B (NF-kappaB) nuclear translocation of vascular endothelial cells induced by burn serum and its correlative mechanism.

METHODSHuman umbilical vein endothelial cells (HUVECs) were cultured in vitro and divided into 5 groups: blank control group (BC, ordinary culture without any stimulation), normal serum control group (NS, cultured with nutrient solution containing 20% healthy human serum), burn serum stimulation group (BS, cultured with nutrient solution containing 20% burn human serum), burn serum+insulin treatment group (BI, cultured with nutrient solution containing 20% burn human serum and 1x10(-7) mol/L insulin), inhibitor pretreatment group [IP, pretreated with 50 micromol/L protein kinase B (Akt) specific inhibitor LY-294002, then cultured with the same medium as used in BI group 30 minutes later] according to the random number table. Six hours later, the injury and apoptosis of HUVECs was respectively observed by the scanning electron microscope and determined by the flow cytometry. Meanwhile, the phosphorylation of inhibitor kappa B-alpha (p-IkappaB-alpha) and Akt (p-Akt) in cytoplasm, and the content of NF-kappaB-p65 in nucleus were determined with Western blot.

RESULTS(1) Compared with those in BC group, HUVECs in BS group shrank obviously with irregular nuclear structure, and intercellular links jagged or vanished. Slight change was observed in HUVECs structure in NS and BI groups, with the cell ductility and nuclear structure much better than those in BS group. (2) The apoptosis rates of HUVECs in BS group [(28.5+/-2.3)%], BI group [(22.3+/-1.8)%], and IP group [(29.7+/-2.4)%] were all obviously higher than that in BC group [(15.7+/-2.2)%, F=14.288, P<0.05 or P<0.01]. There was no significant statistical difference between NS group [(17.0+/-2.5)%] and BC group in apoptosis rate (F=14.288, P>0.05). The apoptosis rate of HUVECs in BI group was obviously lower than that in BS group (F=14.288, P<0.05). (3) Compared with those in BC group, the protein expressions of p-IkappaB-alpha in cytoplasm and NF-kappaB-p65 in nucleus were up-regulated, and the protein expression of p-Akt in cytoplasm was down-regulated in BS and IP groups. The expression levels of the three proteins in NS and BI groups were close to those in BC group.

CONCLUSIONSInsulin could inhibit the IkappaB phosphorylation, and then restrict NF-kappaB nuclear translocation and improve the vascular endothelial cells function accordingly through regulating phosphatidylinositol 3 kinase/Akt pathway.

Apoptosis ; Burns ; blood ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Humans ; I-kappa B Proteins ; metabolism ; Insulin ; pharmacology ; NF-kappa B ; metabolism ; Phosphorylation ; Serum ; metabolism ; Umbilical Veins ; cytology

The objective of this study was to explore the clinical significance of measuring serum free light chains (sFLC) and to compare with serum total light chains (free and binded) in multiple myeloma (MM). sFLC in 20 newly diagnosed MM patients and 20 cases of healthy people as control were measured by immuno-nephelometric assays; the serum light chains and kappa/lambda ratio were measured in all patients, while immunofixation electrophoresis (IFE) tests were carried out at the same time in 18 out of 20 patients. The results showed that the abnormality of serum free light chains and kappa/lambda ratio were found in all of the 20 newly diagnosed MM patients (p < 0.01). The measurement of sFLC showed higher sensitivity than the total serum chains (p < 0.01). It is concluded that the method testing sFLC by immuno-nephelometric assay combined with kappa/lambda ratio is valuable for MM diagnosis, and the measurement of sFLC can be used as one of indicators for MM diagnosis.
Adult ; Aged ; Biomarkers, Tumor ; blood ; Female ; Humans ; Immunoglobulin kappa-Chains ; blood ; Immunoglobulin lambda-Chains ; blood ; Male ; Middle Aged ; Multiple Myeloma ; blood

OBJECTIVETo observe the effect of vacuum sealing drainage (VSD) on the proliferation of Pseudomonas aeruginosa (PA) in infected wound, and to explore its possible mechanism.

METHODSFull-thickness skin wounds each with area of 1 cm x 1 cm were produced on the back of 40 C57 BL/6 mice, and then they were contaminated with wild type PA strains PAO1 marked with target gene of bacterial luciferase luxCDABE (PAO1-lux), they were dressed for 24 hours to reproduce PA infection model. Then mice were divided into experiment [E, with treatment of VSD (pressure value at -16.625 kPa)] and control (C, with treatment of conventional dressing change) groups according to the random number table, with 20 mice in each group. The fluorescence intensity of PAO1-lux and blood flow in wound was respectively measured by in vivo optical imaging system and laser Doppler perfusion imager before treatment and at post treatment hour (PTH) 24. The expression levels of IL-1beta and vascular endothelial growth factor (VEGF) mRNA in wound edge were determined by real-time fluorescence quantitative RT-PCR before treatment and at PTH 24. The specimens of wound edge tissue were collected for observation of pathological change at PTH 24. Data were processed with t test.

RESULTSThere were no obvious difference in fluorescence intensity of PAO1-lux and blood flow in wound between E and C groups before treatment (with t value respectively 0.03, 0.50, P values all above 0.05). The fluorescence intensity of PAOl-lux and blood flow in wound in E group at PTH 24 [(2.69 +/- 0.75) photons x s(-1) x cm(-2) x sr(-1) and (96 +/- 9) PU] was respectively lower and higher than that inC group [(5.18 +/- 0.96) photons x s(-1) cm x (-2) x sr(-1) and (70 +/- 11) PU, with t value respectively 3.54, 3.13, P values all below 0.05]. The expression levels of IL-1beta and VEGF mRNA in both groups before treatment were similar (with t value respectively 0.19, 0.07, P values all above 0.05). The expression levels of IL-1beta and VEGF mRNA in E group at PTH 24 was respectively 4.72 +/- 0.37, 2.68 +/- 0.39, all markedly higher than those in C group (2.24 +/- 0.50, 1.22 +/- 0.13, with t value respectively 6.90, 6.12, P values all equal to 0.00). The number of inflammatory cell infiltrating the wound edge in E group at PTH 24 was increased by nearly 77% as compared with that in C group.

CONCLUSIONSCompared with conventional dressing change, VSD can reduce the amount of Pseudomonas aeruginosa in full-thickness skin defect wound at the early stage, it may be related with an increase in blood flow and number of inflammatory cells in wound tissue, promoting expression of IL-1beta and VEGF mRNA.

Animals ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Negative-Pressure Wound Therapy ; Pseudomonas Infections ; therapy ; Pseudomonas aeruginosa ; Wound Healing ; Wound Infection ; therapy

OBJECTIVETo compare the difference of protein expression in the supernatant of heat injured keratinocytes (KC) and normal KC.

METHODSA model of heat injured KC was produced in vitro. The supernatant of normal KC and heat injured KC was collected after culture for 12 hours, and was ultrafiltered and lyophilized to get the protein. The protein sample was separated by immobilized pH gradient based two dimensional gel electrophoresis (2-DE). The gel was stained and the different expression of protein was analyzed using ImageMaster 2D analysis software.

RESULTS(1) Average protein spots were 1,898 +/- 113, 1,877 +/- 97 in the supernatant of normal and heat injured KC and 1,118 protein spots could be used for statistical analysis. (2) Statistical result showed that 26 protein spots were significantly different between the two groups. 16 protein spots were higher in the supernatant of normal KC and then 10 protein spots were lower in the normal group. (3) 16 protein spots, which included 10 kinds of proteins, were identified successfully as different spots. Lower expression proteins were alpha-enolase, actin cytoplasmic 2, peroxiredoxin-4, phosphoglycerate mutase 1, G protein-regulated inducer of neurite outgrowth l in the supernatant of heat injured KC. Higher expression proteins in heat KC were purine nucleoside phosphorylase, tumor necrosis factor ligand superfamily member 10, proteasome subunit alpha type-7, UDP-glucose 6-dehydrogenase in the supernatant of heat injured KC.

CONCLUSIONSThe result indicated that there are some significant different expression proteins in the supernatant of normal KC and heat injured KC. These findings provide new data for screening major molecules of tissue repair and finding the mechanism of wound repair.

Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; Heat-Shock Response ; Hot Temperature ; Humans ; Keratinocytes ; metabolism ; Proteome ; metabolism

OBJECTIVETo investigate the feasibility of transplanting endothelial progenitor cells (EPCs) transfected with VEGF165 gene to free transplanted fat tissue for increasing neovascularization and the survival.

METHODSEPCs isolated from human cord blood were cultured in vitro and identified by immunocytochemistry. After transfection by VEGF165 gene, the expression of VEGF was assessed using ELISA. Then EPCs with (VEGF gene transfection group) and without VEGF165 gene transfection (EPCs group) were transplanted to free transplanted fat tissue at 18 nude mice's back, and nine nude mice transplanted with free fat tissue were injected with M199 (control group). CM-DiI was used to trace the transplanted cells. The capillary density of transplanted fat tissue was detected by CD34 immunohistochemistry.

RESULTSEPCs expressed cell markers CD34, KDR and CD133. After transfection, the expression of VEGF was positive. Transplanted EPCs survived and proliferated, and transplanted EPCs were incorporated into the capillary networks in the transplanted fat tissue. The percent of survival volume of transplanted fat tissue of VEGF gene transfection group was (96.2 +/- 8.6)%, significantly higher than that of the EPCs group [(75.3 +/- 6.8)%, P < 0.05) and M199 group [(40.2 +/- 2.5)%, P < 0.05). The capillary density of transplanted fat tissue of VEGF gene transfection group was significantly higher than those of the EPCs group and M199 group (P < 0.05).

CONCLUSIONSEPCs from human cord blood can increase free transplanted fat tissue neovascularization and the survival volume, and the ability of promoting neovascularization of EPCs transfected with VEGF165 gene is more potent than EPCs alone.

Adipose Tissue ; transplantation ; Animals ; Cord Blood Stem Cell Transplantation ; Endothelial Cells ; cytology ; physiology ; Female ; Fetal Blood ; cytology ; Graft Survival ; Humans ; Mice ; Mice, Nude ; Stem Cells ; cytology ; physiology ; Transfection ; Transplantation, Heterologous ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; physiology

OBJECTIVETo study the protective effect of intensive insulin treatment on the myocardium of severely scalded rats, and to primarily explore its mechanism.

METHODSEighteen SD rats were divided into three groups, with 6 rats in each group. The rats in burn and intensive insulin group were inflicted with 30% TBSA full-thickness injury on the back. Isotonic saline containing 0.12 U/ml insulin solution, and 100 g/L glucose solution were infused into the rats in the intensive insulin group to keep plasma glucose at the level of 4.0 - 6.6 mmol/L (the total fluid amount was 2 ml x kg(-1) x 8h(-1)). In sham burn group,fluid was given according to physiological demand. The same amount of isotonic saline was infused into the rats in burn group. The venous blood was obtained for the detection of plasma glucose contents, and the left ventricular systolic pressure (LVSP) and left ventricular end-diastolic pressure (LVEDP) were recorded via aortic ventricle cannula before scald and at 1, 2, 3, 4, 5, 6 post-scald hours (PSH). The tissue of the left ventricle was harvested at 6 PSH for the detection of troponin T expression in myocardiocytes.

RESULTSPlasma glucose level was increased to (7.6 +/- 1.7) mmol/L - (8.4 +/- 4.7) mmol/L in burn group during 1-6 PSH, which was significantly higher than that in intensive insulin group (4.5 +/- 0.9) mmol/L - (5.2 +/- 1.3) mmol/L, P < 0.01). Compared with the intensive insulin group, LVSP was markedly decreased in the burn group (60 +/- 11 mm Hg vs 72 +/- 8 mm Hg, P < 0.05) at 1 PSH,whereas LVEDP was increased significantly (21.3 +/- 11.3 mmHg vs 11.7 +/- 5.2 mmHg, P < 0.05). Intensive insulin treatment could significantly inhibit the loss of troponin T protein in myofilaments of myocardium.

CONCLUSIONIntensive insulin treatment possesses a protective effect on myocardia function after severe burns, and it may be related to its preventive effect on the loss of contractile protein in cardiocytes.

Animals ; Blood Glucose ; metabolism ; Burns ; drug therapy ; metabolism ; Insulin ; administration & dosage ; therapeutic use ; Male ; Myocardial Contraction ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Troponin T ; metabolism