Objective To investigate the variation of the chromosomal karyotype of patients with congenital pseudarthrosis of the tibia(CPT) and its relation with the neurofibromatosis. Methods Ten patients with complete follow up records in 28 cases of CPT treated between 1982 and 1999 were included in this study. There were 7 males and 3 females. The age of the patients at the surgery ranged 4 to 17 years. Seven patients had skin caf?-au-lait spots. Peripheral venous blood (1-2 ml) of 10 patients was cultured in 1640 culture medium with 10%(v/v) fetal calf serum and phytahematoagglutinin(PHA) for 70-72 hours, and then colchicines was added (10 ?g/ml) in culture medium 4 hours before finishing the culture. The specimens were harvested and the chromosomal karyotype was analysed. Results The karyotype of the chromosomes were normal(46XY or 46XX) in all of the speciments, there were no chromosome aberration, chromosome loss and polyploid. Conclusion Neurofibromatosis has no relation with CPT, the genic location of the CPT may have some relation with the neurofibromatosis.

Objective To investigate the efficacy of injectable osteoinductive material with fibrin sealant(FS) as a carrier compounded with bovine bone morphogenetic protein(bBMP) and bovine fibroblast growth factor(bFGF) for radial defect in dogs.Methods A total of 12 dogs were used in this study.The animals were randomly divided into treatment and control groups with 6 in each.We created a 20-mm bone defect at the upper radius of each dog,and then sutured the subcutaneous tissues and skin around the lesion.After the operation,FS(control) and FS+bFGF+bBMP were given to the two groups respectively by percutaneous injection.To compare the efficacy of the injections,we examined the animals by radiography in 4,8,16,and 24 weeks.The dogs were sacrificed in 24 weeks to obtain the specimens of the bone defect for histological examination and bone mineral density(BMD) determination.Results Radiography showed callus formation in 4 weeks and then osteoneogenesis in 24 weeks in the treatment group;whereas,in the control group,no callus was found around the defect in 24 weeks.In the treatment group,the mean BMD of the diseased radius was significantly higher than that of the healthy leg and that in the control group [(456.33?13.74) mg/cm2 vs(433.33?6.77) mg/cm2(t=2.57,P=0.00) and 0 mg/cm2].By histological examination,the new-formed bone in the treatment group was confirmed integral and dense with intact cortex and continuous marrow cavities in 24 weeks,while the bone defect of the controls were repaired with connective tissues without remodeling of the bone.Conclusion It is effective to repair bone defect in dogs by using injectable osteoinductive material with FS as a carrier compounded with bBMP and bFGF.

Objective To observe the influence and safety of rosuvastatin on high sensitive C-reactive protein (hs-CRP),Endothelin-1 (ET-1),N-terminal pro-brain natriuretic peptide (NT-proBNP),pulmonary artery systolic pressure(PASP) and cardiac function in patients with chronic pulmonary heart disease.Methods Eighty patients with chronic pulmonary heart disease were enrolled and divided into the statin group(n =40) and the control group (n =40).All patients were given conventional therapy,while the statin group received additionally rosuvastatin 10 mg/d for 6 months.The control group did not receive any lipid-lowering drugs.The plasma levels of hs-CRP,ET-1,NT-proBNP,liver and kidney functions and creatine kinase (CK),echocardiographic indicators of PASP and right ventricular ejection fraction (RVEF) were measured and compared before and after 6-month treatment.Results The levels of hs-CRP,ET-1,NT-proBNP and PASP were significantly lower after 6-month treatment than before treatment in the two groups (Statin group:hs-CRP:(7.45 ± 1.96) mg/L vs.(20.67 ± 5.12) mg/L,t =9.57,P ＜ 0.01 ; ET-1:(45.72 ± 6.85) ng/L vs.(56.39 ±7.34) ng/L,t =3.78,P ＜ 0.01 ; NT-proBNP:(136.54 ± 20.67) ng/L vs.(182.83 ± 23.27) ng/L,t =4.15,P ＜0.01 ;PASP:(42.6 ± 6.3)mm Hg vs.(52.3 ± 8.4) mm Hg,t =3.54,P ＜ 0.01 ; Control group:hs-CRP:(12.73 ±3.14) mg/L vs.(20.58 ±4.98)mg/L;t =4.96,P ＜0.01 ;ET-1:(51.66 ± 6.42)ng/L vs.(56.43 ±7.81) ng/L,t =3.43,P ＜ 0.01 ; NT-proBNP:(162.74 ± 21.59) ng/L vs.(181.56 ± 22.78) ng/L; t =3.60,P ＜0.01 ;PASP:(45.7 ±6.5) mm Hg vs.(51.8 ± 8.2) mm Hg,t =3.62,P ＜ 0.01),but the statin group reduced even more significantly (t =2.36,2.21,2.25 and 2.09 respectively,P ＜ 0.05).The level of RVEF was significantly higher after 6-month treatment than before treatment in the two groups (Statin group:(50.8 ±7.9) ％ vs.(41.5 ±6.7)％,t =3.69,P ＜0.01 ;Control group:(46.6 ±7.8)％ vs.(42.0 ±6.2)％,t =3.58,P ＜ 0.01),but the statin group increased even more significantly(t =2.18,P ＜ 0.05).Statistical differences of liver and kidney function and serum CK were not found in the two groups before and after treatment(P ＞ 0.05).The adverse reaction in the statin group was few.Conclusion Rosuvastatin can reduce the levels of hs-CRP,ET-1,NT-proBNP and PASP,improve RVEF and cardiac function in patients with chronic pulmonary heart disease,and its security is fine.

Objective To study the effect of injectable osteoinductive materials with fibrin sealant as the vehicle on the reconstruction of segmental radius defect in dog, and to provide an experimental foundation for its clinical application in the future. Methods 20mm bone defects were created in the radius 18 dogs, and in the experiment group b [FS+bFGF+bBMP (bovine BMP)] composite, and in experiment group r (FS+bFGF+rhBMP-2) were injected percutaneously into the defects. A control group (FS) with bone defect only was also included. The reconstruction effect on the segmental dog radius defects was evaluated by radiography, histology, bone mineral density (BMD) at 4, 8, 12, 16, 20 and 24 weeks after operation. Results In the experimental groups with bovine BMP as osteoinductive agent, higher repairing capability and osteogenesis rate were observed in experiment group b (FS+bFGF+bBMP) compared with that of control group (P

Objective To investigate the effect of the injectable osteoinductive material with fibrin glue(FG)as carrier compounded with rhBMP-2 and bFGF on the proliferation and differentiation of marrow matrix cells(MMCs)of rabbit,in order to lay an experimental foundation for clinical application in the future.Method MMCs were isolated and cultured from bone marrow of a 3-day-old rabbit.The effects of the material were investigated in experiment group(group A:FG+bFGF+rhBMP-2),with control groups consisting of control group 1(group B:FG),control group 2(group C:FG+bFGF),control group 3(group D:FG+rhBMP-2)and single control group(group E:free of material).The observation objects included proliferation rate,adhesive rates,expression of type I collagen and alkaline phosphatase,cell growth condition in the material and the ultrastructure of rabbit MMCs with the aid of electron microscopy,histochemistry and cell culturing.The concentration of rhBMP-2 and bFGF in all groups was uniform(1?g/ml).Results The proliferation rates and the adhesive rates of rabbit MMCs in group A was significantly higher than that in group B,group D and group E,but was lower than in group C(P

BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.

OBJECTIVE: To study the effects of Wenyang Chubi Decoction (WYCBD), a compound traditional Chinese herbal medicine, on connective tissue growth factor (CTGF) and collagen-I (COL-I) in a mouse model of scleroderma. METHODS: Scleroderma was induced in BALB/c mice by daily local injection of bleomycin for three weeks and the mice were randomly divided into untreated, WYCBD-treated and normal saline (NS) treated groups, with another group of BALB/c mice as normal control. WYCBD and NS were given orally for one month respectively. Histopathology in the skin and lungs of the mice were examined. The CTGF and COL-I expressions in the skin or skin lesions were detected by immunohistochemical Elivision assay. RESULTS: The expression levels of CTGF and COL-I in the untreated group were significantly higher than those in the normal control group (P<0.05). Compared with the NS-treated group, the WYCBD-treated group had significant improvement in the skin and lung histopathology and remarkably decreased expression levels of CTGF and COL-I (P<0.05). CONCLUSION: Scleroderma mice showed high expressions of CTGF and COL-I in the skin. WYCBD had the effects of decreasing the CTGF and COL-I expressions and improving the skin fibrosis.

Objective We reported previously that single nucleotide polymorphisms SNP) + 11G ＞ A in intron 3 of the human urate transporter 1 (hURAT1) gene are associated with hyperuncaemia in Han Chinese.The aim of the present study was to evaluate the effect of the variants on hURAT1 function.Methods The wild-type,mutant-type hURAT1 and exon 5-null hURAT1 were constructed,and respectively microinjected into the zebrafish embryo yolks.The subcellular localization of different genotypes of hURAT1 was detected by confocal laser scanning microscope.Results Compared with wild type,the mutant recombinant plasmid transcribed two types of mRNA spliceosome,the wild type and the exon 5-null type.The hURAT1 wild type protein was prominent localized on cell membrane,while the mutant type and exon 5-null hURAT1 proteins were distributed uniform in the cytoplasm but not on the cell membrane.Conclusion The hURAT1 variant + 11 G ＞ A resulted in an alternative splicing of hURAT1 mRNA-exon 5-null type.Its protein product exhibited a different subcellular localization compared with that of wild type.

Objective To explore the influence of injury severity on transplantation of embryonic neural stem cells (NSCs) after traumatic brain injury (TBI).Methods The NSCs were isolated from the hippocampus of fetal rats aged at from 12-14 days.The cells were cultured and proliferated in the serum-free medium and identified in vitro.The animals received transplants in the bilateral hippocampal areas at day 3 following mild or moderate TBI separately.Conventional histology,TUNEL and immunohistology were examined to detect BrdU,NSE,GFAP,GalC,NGF and BDNF at day 14 post-implantation.Results BrdU-labeled positive cells in the bilateral hippocampus in the mild TBI group were more than those in the moderate TBI group at day 14 post-implantation.Significant differentiation of the astrocytes recognized as GFAP positive cells in the bilateral hippocampus was found at day 14 post-implantation.The expression of NGF and BDNF proteins was increased following TBI,the most evident in the mild TBI group.Conclusion The influence of injury severity on transplantation may be associated with the change of the microenvironment after TBI.

BACKGROUND:Different methods to remove immunogenicity have different effects on the spatial microstructure of antigen-extracted heterologous bone.
OBJECTIVE:To compare the microstructure of the antigen-extracted heterologous bone via different methods to provide experimental data for tissue engineering industrialization.
METHODS:Fresh cancellous bones extracted from adult sheep vertebrae were prepared into cylinders. Their long axis direction was the same with orientation of the trabeculae. After vibration washing and different-frequency ultrasound rinsing, the cylinder samples were randomly divided into three groups:in physical calcined group, the samples were defatted, decellularized and deproteinized sequential y using methanol/chloroform and hydrogen peroxide, then bathed in sodium pyrophosphate and directly calcined at 1 000 ℃ for 3 hours;in chemical group, the samples were defatted, decellularized and deproteinized sequential y using methanol/chloroform and hydrogen peroxide;in control group, the samples were dried natural y at room temperature. Microstructure of the samples in each group was analyzed and compared through determination of porosity, scanning electron microscopy observation, X-ray diffraction analysis, X-ray atomic spectroscopy elemental analysis microscopic spatial structure.
RESULTS AND CONCLUSION:The physical calcined and chemical groups maintained natural network pore system to different extents. The size of the large pore was 50-600μm and that of the smal one was about 2μm. The porosity was 55%to 70%. Hydroxyapatite was the main component of the physical calcined group which was determined by X-ray diffraction, and a smal amount of theβ-Tricalcium phosphate was also determined. In the chemical group, the main component was only hydroxyapatite. The three-dimensional spatial structures of the deproteinized cancellous bones were not damaged greatly, and they had a natural pore network system. Antigen component of xenogeneic cancellous bone can be more thoroughly removed by physical calcination step. The scaffold material made by antigen-extracted heterologous bone may satisfy the demands for bone tissue-engineering scaffolds.