Relating surface roughness and magnetic domain structure to giant magneto-impedance of Co-rich melt-extracted microwires.

Understanding the relationship between the surface conditions and giant magneto-impedance (GMI) in Co-rich melt-extracted microwires is key to optimizing their magnetic responses for magnetic sensor applications. The surface magnetic domain structure (SMDS) parameters of ~45 µm diameter Co69.25Fe4.25Si13B13.5-xZrx (x = 0, 1, 2, 3) microwires, including the magnetic domain period (d) and surface roughness (Rq) as extracted from the magnetic force microscopy (MFM) images, have been correlated with GMI in the range 1-1000 MHz. It was found that substitution of B with 1 at. % Zr increased d of the base alloy from 729 to 740 nm while retaining Rq from ~1 nm to ~3 nm. A tremendous impact on the GMI ratio was found, increasing the ratio from ~360% to ~490% at an operating frequency of 40 MHz. Further substitution with Zr decreased the high frequency GMI ratio, which can be understood by the significant increase in surface roughness evident by force microscopy. This study demonstrates the application of the domain period and surface roughness found by force microscopy to the interpretation of the GMI in Co-rich microwires.

SHH-dependent knockout of HIF-1 alpha accelerates the degenerative process in mouse intervertebral disc.

Hypoxia-inducible factor-1alpha (HIF-1 alpha) has been reported to have an important role in the metabolism and synthesis of extracellular matrix of the nucleus pulposus cells (NPCs) and was assumed to be involved in the process of intervertebral disc degeneration. The objective of this study was to investigate the role of HIF-1alpha in disc degeneration in vivo using a conditional HIF-1alpha knockout (KO) mouse model. ShhCre transgenic mice were mated with HIF-1 alpha fl/fl mice to generate conditional HIF-1alpha KO mice (HIF-1alpha fl/fl-ShhCre+). Three mice of each genotype (Wide-type and HIF-1alpha KO) at the age of 3 days, 6, and 12 weeks were sacrificed after genotyping. Five lumbar disc samples were harvested from each mouse, with a total of 45 disc samples for each genotype. In situ hybridization and immunohistochemical analysis were used to check the efficacy of HIF-1alpha knockout. Histological grading of the disc degeneration was performed according to the classification system proposed by Boos et al. Picro-sirius red staining, Safranine O/fast green staining and immunohistochemical study were used to evaluate the expression of aggrecan, type-II collagen and vascular endothelial growth factor (VEGF). Histologic analysis revealed more NPC deaths and signs of degeneration in HIF-1alpha KO mice and the degeneration scores of HIF-1alpha KO mice were significantly higher than those of the Wide-type mice at the age of 6 weeks and 12 weeks. There were less expressions of aggrecan, type-II collagen and VEGF in the intervertebral discs of HIF1-alpha KO mice than in those of wild-type mice. Taken together, the results of our study indicated that HIF-1alpha is a pivotal contributor to NPC survival and the homeotasis of extracellular matrix through the HIF-1alpha/VEGF signaling pathway, and plays an important role in the development of disc degeneration.

Reduced expression of insulin-like growth factor 1 receptor leads to accelerated intervertebral disc degeneration in mice.

Insulin-like growth factor 1 (IGF-1) and its receptor (insulin-like growth factor 1 receptor, IGF1R) can regulate the extracellular matrix synthesis and play a crucial role in maintaining the normal functions of the intervertebral disc (IVD). The objective of this study was to investigate whether there would be accelerated IVD degeneration (IVDD) in IGF1R+/- mice. Three IGF1R+/- male mice and three wild-type male mice were sacrificed respectively at 6, 12, and 18 weeks after birth. Six lumbar disc samples were harvested from each mouse, with a total of 54 disc samples taken from each genotype. Histomorphological analysis for the IVD was performed to assess the degenerative extent according to the classification system proposed by Boos et al. Quantitative real-time PCR and semi-quantitative histologic scoring (HScore) for immunohistochemical staining were used to evaluate the expression level of type-II collagen, aggrecan and matrix metallopeptidase 13 (MMP-13). Histomorphological analysis for the discs revealed significantly less amounts of proteoglycan and type-II collagen, and significantly higher total degenerative score in IGF1R+/- mice than in wild-type mice. Real-time PCR showed that the mRNA expressions of type-II collagen and aggrecan in the discs were significantly lower, while MMP-13 was significantly higher in IGF1R+/- mice than in wild-type mice. The results of HScore analysis were similar to those obtained from the quantitative real-time PCR. Taken together, our study indicates that reduced expression of IGF1R would lead to accelerated degeneration of IVD. IGF1R+/- mice could be regarded as a good animal model to study IVD degeneration (IVDD), and studies on the IVD of IGF1R+/- mice could provide further insight into the pathogenesis of IVDD.

Differential responsiveness to 17ß-estradiol of mesenchymal stem cells from postmenopausal women between osteoporosis and osteoarthritis.

UNLABELLED: Differential osteogenic potential and responsiveness to 17ß-estradiol (E2) of mesenchymal stem cells (MSCs) were found between postmenopausal women with osteoporosis (OP) and osteoarthritis (OA). These results suggest differential biological mechanisms of estrogen deficiency in regulation of bone remodeling between OP and OA. INTRODUCTION: OP and OA are two common disorders in postmenopausal women. The inverse relationship has been suggested between OP and OA, but their mechanisms that relate to estrogen deficiency are not fully understood. The aim of this study was to compare the differential responsiveness to E2 of MSCs from osteoporotic versus osteoarthritic donors. METHODS: Twenty postmenopausal patients, ten with osteoporotic hip fractures and ten with hip osteoarthritis, were included into this study. MSCs were derived from cancellous bones of femoral heads from OA and OP donors and cultured in osteogenic and adipogenic medium with or without E2 added. The alkaline phosphatase (ALP) activity, calcium content, calcified nodules, lipid droplets, messenger RNA (mRNA) expression of ALP, osteocalcin (OC), collagen 1α (COL1α), peroxisome proliferators-activated receptor γ2 (PPARγ2) and lipoprotein lipase (LPL) were measured and compared between two groups with OP and OA. RESULTS: In osteogenic medium, ALP activity, calcium content and mRNA expression of OC and COL1α in MSCs from OA were significantly higher than those from OP group. In adipogenic condition, there was no significant difference in lipid droplets formation and mRNA expression of PPARγ2 and LPL between OP and OA groups. With E2 added in osteogenic medium, ALP activity, calcium content and OC mRNA were significantly higher in OP group than in OA group, whereas E2 had no significant effect on lipid droplet formation and mRNA expression of PPARγ2 and LPL. CONCLUSION: Differential osteogenic potential and responsiveness to E2 of MSCs were found between postmenopausal women with OP and OA. These results may provide information for clinical application of MSCs in the differential setting of estrogen deficiency.

Bone loss and impaired fracture healing in spinal cord injured mice.

UNLABELLED: Spinal cord injury (SCI) results in impaired fracture healing in mice while leading to significant bone loss. Poor fracture healing following SCI is consistent with significant bone loss. INTRODUCTION: SCI leads to significant bone loss in sublesional limbs, but there is few data concerning the relationship between fracture healing and bone loss following SCI. This study was undertaken to investigate the effect of SCI on fracture healing using a mouse femur fracture model. METHODS: One hundred twenty male C57BL/6J mice were randomly divided into SCI and control groups (n=60, respectively). A femoral shaft fracture was generated and fixed with intramedullary pins 3 weeks after SCI. Fracture healing was evaluated by micro-computed tomography (micro-CT) for callus formation and mineralization and neovascularization, and bone mineral density (BMD) was measured by DXA at 1, 2, and 4 weeks after fracture. Serum vascular endothelial growth factor (VEGF), osteocalcin, and alkaline phosphatase (ALP) were assessed using ELISA at each time point. Biomechanical testing was performed at 2 and 4 weeks. RESULTS: BMD in SCI mice was significantly lower compared to control mice at each time point, with callus volume and all vessel parameters reduced as measured by micro-CT. Ultimate stress of the femora was significantly lower in SCI mice than in control mice at 2 and 4 weeks after fracture, whereas Young's modulus between the SCI and control mice turned to be significantly different at 4 weeks. Serum VEGF was lower in SCI mice than in the control group at 2 and 4 weeks, whereas serum osteocalcin and ALP were lower in SCI mice than in control ones at each time point. CONCLUSION: Significant bone loss and fracture healing impairment was noted in SCI mice. Decreased angiogenesis is consistent with the changes of microarchitecture and biomechanical properties during fracture healing.

Micro-CT and mechanical evaluation of subchondral trabecular bone structure between postmenopausal women with osteoarthritis and osteoporosis.

SUMMARY: An inverse relationship between osteoarthritis and osteoporosis has been debated over years. The microstructure of the femoral heads from postmenopausal osteoarthritic and osteoporotic women was evaluated with micro-CT. Significant differences were observed in microstructural parameters between them. Different microstructure might relate to the opposite bone defects in osteoarthritis and osteoporosis. INTRODUCTION: This study was undertaken to verify the inverse relationship between osteoarthritis (OA) and osteoporosis (OP) by comparing the structural and mechanical indices. METHODS: Femoral head specimens were obtained from 17 postmenopausal women (OA, n = 8; OP, n = 9) during hip surgery. The microstructural parameters were measured with micro-CT. Mechanical test was performed after bone cube scanning. RESULTS: Significant difference in bone volume fraction (BV/TV) and trabecular thickness was noted between OA and OP groups. Structure model index decreased in OA, and increased in OP. The higher apparent density (AD) and lower material density (MD) were also shown in OA. Different from OP, positive correlation were noted between connectivity density and mechanical indices in OA. In OA group, BV/TV was associated with Young's modulus and AD, while trabecular number was the only parameter that correlated with MD. However, in OP group, only BV/TV correlated with yield strength, AD, and MD. CONCLUSIONS: We observed the difference in microstructure between postmenopausal women with OA and OP, which might relate to the opposite bone defects in OA and OP. BV/TV might play an important role in mechanical properties of the subchondral bone in either OA or OP.

Changes in bone mass, bone structure, bone biomechanical properties, and bone metabolism after spinal cord injury: a 6-month longitudinal study in growing rats.

Spinal cord injury (SCI) results in a great decline in bone mineral density (BMD) and deterioration of bone microarchitecture. The objective of this study was to investigate the time course of the changes in BMD, microarchitecture, biomechanical properties, and bone turnover in male growing rats following SCI. Sixty male growing Sprague-Dawley rats, 6 weeks of age, were randomly divided into SCI (lower thoracic cord transection) and sham-operated groups, and bone tissues and blood samples were examined at 3 weeks, 6 weeks, and 6 months after surgery. SCI rats had low bone weight (wet, dry, and ash weight) and BMD of the femora, tibiae, and third lumbar vertebrae at all time points compared to sham rats, while in forelimbs, there was a decrease of dry and ash weight compared to sham rats only at 3 weeks but not BMD. Bone microarchitecture and trabecular connectivity were deteriorated in SCI rats and remained so after. Bone formation rate and serum osteocalcin level in SCI rats were significantly increased 3 weeks after SCI surgery. However, eroded surface/bone surface and serum N-terminal telopeptide of type I collagen level remained elevated from 3 weeks to 6 months. In addition, SCI rats showed poor biomechanical properties in the proximal tibiae and femora but not in the humeri. In conclusion, SCI causes profound BMD loss, disturbances in bone microarchitecture, decreased mechanical strength in the lower extremity and lumbar spine, and high bone turnover. These findings will allow better understanding of osteoporosis following SCI.

Differences of bone mass and bone structure in osteopenic rat models caused by spinal cord injury and ovariectomy.

UNLABELLED: Both spinal cord injury and ovariectomy can result in ostepenia in rats. SCI induces more deterioration of cortical geometric structure and trabecular microstructure in the proximal tibial metaphysis than OVX. The proximal tibial metaphysis microstructure significantly correlates with its biomechanical properties. INTRODUCTION: The purpose of the present study was to compare the effects of spinal cord injury (SCI) and ovariectomy (OVX) on bone gain in young female rats. METHODS: Thirty young female Sprague-Dawley rats were randomized into three groups: age-matched intact control (CON), OVX and SCI. The tibiae were assessed for DXA and micro-CT analysis, biomechanical testing, the upper tibial epiphyseal plate height, and blood samples for biochemical analysis. RESULTS: SCI rats showed lower aBMD in the proximal tibiae as compared with OVX rats. Cortical geometric structural parameters of the tibial midshaft in SCI rats were significantly lower than OVX rats. SCI or OVX induced significant changes in all trabecular microstructural parameters in the proximal tibial metaphysis. The trabecular separation (Tb.Sp) and structure mode index (SMI) in SCI rats were significantly higher than in OVX rats. BV/TV explained 84% of the variation of ultimate load of the proximal tibial metaphysis. There was no difference of the upper tibial epiphyseal plate height between SCI and OVX rats. Serum NTX level in SCI rats was significantly higher than in OVX rats. CONCLUSIONS: SCI induces more deterioration of cortical bone geometric structure and trabecular microstructure in the proximal tibial metaphysis than OVX.

Effects of spinal cord injury on osteoblastogenesis, osteoclastogenesis and gene expression profiling in osteoblasts in young rats.

INTRODUCTION: Spinal cord injury (SCI) causes a significant amount of bone loss in the sublesional area in animals and humans, and this type of bone loss is different from other forms of osteoporosis such as disuse osteoporosis and postmenopausal osteoporosis. However, no data is available on the cellular and molecular changes of osteoblastogenesis and osteoclastogenesis during SCI-induced bone loss. METHODS: SCI and SHAM rats were used in this study to investigate osteoblastogenesis and osteoclastogenesis in bone-marrow culture. We also measured bone mass and bone histomorphometry, as well as the expression of alkaline phosphatase (ALP), core binding factor alpha1 (Cbfa-1), osterix, receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG) in osteoblast-like cells in bone-marrow culture obtained from SCI and SHAM rats. RESULTS: Bone mineral density (BMD) measurement showed serious bone loss in the tibial ephiphyses and metaphyses of SCI rats compared with SHAM rats. In addition, bone histomorphometry analysis of the tibial metaphyses of SCI rats demonstrated that bone microarchitecture in SCI rats deteriorated further than in SHAM rats, and increased eroded surfaces and bone formation rates were observed in SCI rats. The number of osteoclasts that developed from bone marrow of SCI rats at equal density was significantly increased compared with SHAM rats, and the area of the resorption pits formed in the bone marrow culture from SCI rats was significantly greater than SHAM rats, whereas the number of CFU-F and CFU-OB was similar in both groups. RANKL mRNA and protein levels in osteoblast-like cells in culture obtained from SCI rats were significantly higher than those from the SHAM rats, whereas OPG levels decreased slightly. The ratios of RANKL to OPG expression in SCI rats were significantly higher than those in SHAM rats. However, osteogenic gene profiling of Cbfa-1, ALP and osterix in SCI rats remained similar with SHAM rats. CONCLUSION: These changes favor increased osteoclast activity over osteoblast activity, and may explain, in part, the imbalance in bone formation and resorption following SCI.

Spinal cord injury causes more damage to bone mass, bone structure, biomechanical properties and bone metabolism than sciatic neurectomy in young rats.

INTRODUCTION: Although both spinal cord injury (SCI) and sciatic neurectomy (NX) can cause osteopaenia in young rats, the effects of these two injuries on cortical and cancellous bone may differ. The objective of this study was to compare the effects of SCI and NX on bone weight, bone material property, bone mass, bone geometry, trabecular microarchitecture, mechanical strength and bone turnover in young rats. MATERIALS AND METHODS: Thirty six-week-old male Sprague-Dawley rats were randomised into three groups (10 per group): SCI, bilateral sciatic NX and untreated control (CON). All rats were killed on day 21. Bone mineral density (BMD) was studied using dual-energy X-ray absorptiometry (DXA). At death, the right proximal tibial metaphysis and the fourth lumbar vertebra were examined for bone structural geometric analysis by micro-computed tomography (CT) and then processed for histomorphometry to assess bone cell activity. Serum N-terminal telopeptide of type I collagen (NTX) and osteocalcin (OC) levels were analysed by enzyme-linked immunosorbent assay (ELISA). Biomechanical strength properties of the femur and humerus were measured by three-point bending, and the third lumbar vertebra and the proximal end of tibia were tested by compression. RESULTS: BMD in the sublesional areas of SCI rats was significantly lower than that of NX rats (proximal tibia, 0.176+/-0.018 g/cm(2) vs. 0.224+/-0.015 g/cm(2), P<0.001). Bone volume (BV/TV), trabecular number (Tb.N) and thickness (Tb.Th) in the tibial second spongiosa of SCI rats were significantly less than those in NX rats (BV/TV: 7.15+/-1.18% vs. 12.32+/-1.83%, P<0.001; Tb.N: 1.23+/-0.22 vs. 2.38+/-0.45, P<0.001; Tb.Th: 33.73+/-5.15 microm vs. 42.80+/-7.44 microm, P<0.01) and trabecular separation (Tb.Sp: 1,053.37+/-164.24 microm vs. 748.32+/-129.36 microm, P<0.01) was significantly greater than in NX rats. Furthermore, poorer trabecular connectivity was found in SCI rats than in NX rats (number of nodes, N.Nd/TV: 1.04+/-0.09 vs. 3.29+/-0.53; number of terminus, N.Tm/TV: 28.53+/-3.17 vs. 21.64+/-2.31, P<0.01). The bone formation rate of the tibial second spongiosa in SCI rats was significantly higher than in NX rats (2.06+/-0.13 vs. 1.53+/-0.09, P<0.001) and, also, the eroded surface in SCI rats was significantly higher than in NX rats (13.42+/-1.24 vs. 10.36+/-1.07, P<0.001). In addition, biomechanical tests showed that SCI rats had poorer biomechanical properties of the femur, proximal tibia and fourth lumbar vertebra than in NX rats. There were significantly higher levels of OC in SCI rats compared with NX rats (30.19+/-1.17 vs. 21.15+/-1.76, P<0.001). Also, serum NTX levels were significantly higher than in NX rats (51.60+/-2.61 vs. 33.85+/-1.93, P<0.001). CONCLUSION: SCI caused more damage to bone mass, bone structure, biomechanical properties and bone metabolism than NX in young rats. This suggests that different mechanisms may underlie osteopaenia following SCI and NX.

The tumor suppressor adenomatous polyposis coli gene is associated with susceptibility to schizophrenia.

The etiology of schizophrenia is unclear, although family, twin, and linkage studies implicate genetic factors. Here, we identified adenomatous polyposis coli (APC), a tumor suppressor gene, as a risk factor for schizophrenia. We compared leukocytic gene expression patterns of six pairs of patients with schizophrenia and healthy controls by microarray. APC expression levels were significantly increased in all patients compared to healthy controls. To confirm the findings of microarray analysis, we measured expression levels of APC in the leukocytes from 30 relapse patients taking antipsychotic medication, 29 first-episode drug-naïve patients, and 30 healthy controls using real-time quantitative reverse transcription (RT)-polymerase chain reaction (PCR). APC expression levels were significantly increased in leukocytes of schizophrenics both taking and not taking antipsychotic medication and hence the increase of APC expression was not due to antipsychotic medication. APC is located at 5q21-22, which has been previously reported to be linked with schizophrenia. Further, we performed the transmission disequilibrium test (TDT) and TDT based on haplotypes to search for the association between schizophrenia and APC by examining 163 parent-offspring trios of Chinese descent. We analyzed three single-nucleotide polymorphisms (SNPs) (rs2229992, rs42427, rs465899) at the exon region of APC. TDT showed that the three SNPs are significantly associated with schizophrenia (TDT chi(2)=4.23, P<0.05; 4.15, P<0.05; 8.49 P<0.01, respectively; HHRRchi(2)=5.54, P<0.05; 4.40, P<0.05; 9.79, P<0.01, respectively). We found a significant association between the APC haplotypes from rs2229992-rs42427-rs465899 and schizophrenia (Global chi(2)=44.376,df=7, P<0.001). The C-A-T haplotype has a frequency of more than 57% and has a strong association with schizophrenia (chi(2)=15.04, P<0.001). These results indicate that the APC may be a candidate gene conferring susceptibility to schizophrenia and also may be associated with reduced vulnerability to cancer in schizophrenia.

Detection of infectious hepatitis A virus by integrated cell culture/strand-specific reverse transcriptase-polymerase chain reaction.

AIMS: A novel integrated cell culture/strand-specific reverse transcriptase-polymerase chain reaction (RT-PCR) assay was established for detection of infectious hepatitis A virus (HAV). METHODS AND RESULTS: The specificity of tagged RT-PCR was assessed using HAV genomic positive-strand RNA extracted from HAV virions as reference. Water samples artificially contaminated with infectious or formalin-inactivated HAV were subjected to integrated cell culture (ICC)/RT-PCR and ICC/strand-specific RT-PCR assays respectively. The tagged RT-PCR had high specificity for HAV negative-strand RNA. By demonstrating the formation of negative-strand RNA replicative intermediate, ICC/strand-specific RT-PCR can distinguish between infectious and non-infectious HAV. The described method detected infectious HAV at inoculation level of 10(0) TCID50 per flask within 4 days. CONCLUSIONS: The ICC/strand-specific RT-PCR is a novel, rapid, sensitive and reliable method for detection of infectious HAV. SIGNIFICANCE AND IMPACT OF THE STUDY: Coupled with a suitable virus concentration and purification system, ICC/strand-specific RT-PCR will provide a novel and rapid method for detection of infectious HAV in clinical, environmental and food samples. This assay may be used as an alternative method to test the effective inactivation of inactivated virus vaccines. It may also be adapted to assess the efficacy of disinfection of HAV and enteric viruses in foods and water.

A case of acute eosinophilic granulocytic leukemia with PML-RAR alpha fusion gene expression and response to all-trans retinoic acid.

A typical case of eosinophilic granulocytic leukemia with PML-RAR alpha fusion gene expression is reported. The patient achieved complete remission after oral administration of all-trans retinoic acid without any exposure to cytotoxic agents. The facts strongly suggest that the genetic event occurred at the level of pluripotent stem cells.

[Trial manufacture and application of soft uterine probe with reading scales (Type III) for the detection of IUD position].

PIP: The position of the IUD in the uterus is important to its contraceptive effectiveness. Pelvic x-ray examination is often imprecise and can not reveal whether the IUD is lodged in the muscles or outside of the uterus. Ultrasonic examination requires expensive equipment and is a complicated procedure. Examination of the position of an IUD inside the uterus with a flexible probe may solve the problem of uterine perforation which occurs with the use of a metal probe and may also reduce the incidence of unnecessary extraction and allow an appropriate correction of the IUD position. In August 1985, the soft silicone rubber uterine probe with a reading scale was developed and manufactured by the No. 3 Hospital of China's Medical University. The probe is inserted into the uterus together with the IUD. After the insertion procedure, clients walk about 500 meters, and a side-ways x-ray or radiography examination of the position of the IUD and the probe shows whether the IUD is properly placed or descended. A clinical study was conducted with 1000 IUD users, including 222 cases of IUD insertion after abortion and 162 cases of uterine bleeding it IUD in situ. A comparison of the post-abortion group, the irregular bleeding group and the normal group shows that a larger proportion of the former 2 groups had experienced an IUD descending. The results suggests that the large size of a post-abortion uterus makes it hard for an IUD to stay in place. Improper positioning of the IUD may be the cause of the irregular bleeding. The soft probe may also be used for uterine examinations. In cases of malformation of the uterus or of pelvic tumor, the use f the soft probe under ultrasonic examination may confirm the position of the uterus or tumor. The soft probe can also be used to clarify the association of irregular bleeding with the IUD.^ieng

Inactivation of poliovirus with beta-propiolactone.

The recovery of poliovirus D-antigen after virus inactivation was studied for two inactivating agents (beta-propiolactone and formalin) using the three poliovirus types (Sabin types 1, 2 and 3). With beta-propiolactone (BPL), D-antigen recoveries were high (88, 88 and 60%, respectively) but were significantly less when formalin was used (22, 15 and 25%). beta-Propiolactone inactivated virus was purified, combined with Freund's adjuvant and used to hyperimmunize rabbits. High titres (50 000-200 000) of specific neutralizing antibody were obtained.

A simple endpoint dilution method for evaluating serum used for cell culture.

A simple endpoint dilution method for evaluating foetal calf serum quality is described. The test uses a series of doubling dilutions of cells on microtitre trays with the test sera added to replicate dilution series. After five to six days of incubation the cells are stained with crystal violet and the end points read macroscopically. The cell growth-promoting property of serum may be expressed as a reciprocal of the cell dilution resulting in an approximately 50% coverage of cells.