OBJECTIVETo observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers, such as alkaline phosphatase, Cbf(alpha)l and type I collagen, and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head.

METHODSFifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs, then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood, cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group, model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro, and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week,the expressions of alkaline phosphatase, Cbf(alpha)l and type I collagen mRNA were detected.

RESULTSThe alkaline phosphatase activity was significantly decreased in BMSCs of the model group as compared with the blank group (P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly increased in BMSCs of each HSYA group as compared with the model group (P < 0.05 or P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also increased significantly (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chalcone ; analogs & derivatives ; chemistry ; pharmacology ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor alpha Subunits ; genetics ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rabbits

Purpose:An analysis of 27 patients who underwent surgery for their malignant fibrous histiocytoma in the soft tissue of the neck was done to show that compartment dissection of the neck should be the optimal choices of treatment. Methods:From 1 st Jan 1972 through 31 st Dec 1993,27 patients who were referred to the Cancer Hospital for surgical treat- ment were reviewed,including 19 males and 8 females in the age range from 15 to 67,with a mean of 44 years.Surgery was classified as wide resection and compartment dissection of the neck.The two groups were compared for local recurrence and 5 years survival rates.Results:Of the 27 operations,21 were wide resections and 6 compartment dissection of the necks.The average tumor diameter was6 millimeters in patients with compartment dissection of the neck.Local recurrence was encountered in 16 of 21 wide resections (76.2%),and in 1 of 6 compartment dissection of the necks (16.7%).The difference between the two groups was statistically significant (P= 0.015).The 5 years survival was 89% for patients without local recurrence and 52% for patients with local recurrence. The difference between them was significant statistically (Log Rank,P=0.0264).Conclusions:For malignant fibrous his- tiocytoma in the soft tissue of the neck,compartment dissection of the neck is recommended as the optimal operation for its ability to the local control of the disease locally.

Background Researches demonstrated that ciliary neurotrophic factor (CNTF) can enhance survival and promote differentiation of neutron.Meanwhile,CNTF also is thought to play an important role in the development of visual pathway.But,less studies are reported in the relationship of CNTF and form deprivation amblyopia.Objective This study was to investigate the expressions of CNTF in visual cortex area 17 in form deprivation amblyopia model.Methods Twelve 4-week-old cats were randomized into normal group and form deprivation amblyopia group.Monocular form deprivation amblyopic models were established in 6 cats by eyelids suture method.Pattern visual evoked potential(P-VEP) was recorded to evaluate the amblyopic models 16 weeks later following the eyelids suturing.Then,bilateral visual cortex tissue was incised at a vertical in sagittal axis fashion to prepare the section.Nissl staining was used to detect the morphologies of neurons.Expression of CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 was located and quantified by immunochemistry.The positive cell number and gray scale for CNTF were calculated and compared between two groups.The use of the animals complied with Regulations for the Administration of Affairs Coucerning Experimental Animals by State Science and Technology Commission.Results Compared with the normal group,P-VEP amplitude was significantly reduced (6.11 ±1.56 μV vs.11.42±t.92 μV) and latency was significantly prolonged(75.77±9.83 ms vs.58.56±7.17 ms) in the form deprivation amblyopia group (t=5.272,3.464,P＜0.05).Nissl staining showed that the number of neurons in the form deprivation amblyopia group was less than that in the normal group.In the form deprivation amblyopia group,neurons became shrinkage and turned round,cytoplasmic processes get shortened,and the nucleus became small.The number of Nissl bodies was decreased.lmmunochemistry showed the positive neutrons for CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 in hoth normal cats and model cats with the more positive cells in Ⅱ-Ⅳ layers.Compared with the normal group,the positive cell number for CNTF was significantly reduced in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group (Ⅱ layer:95.93±8.24 vs.116.25±6.52;I layer:102.65±7.45 vs.125.23±8.21;Ⅳ layer:l10.65±6.85 vs.139.54±4.26) (t=4.737,4.989,8.773,P＜0.05).In addition,the gray scale of CNTF positive cells was significantly lower in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group than that the normal group (Ⅱ layer:106.98 ± 8.86 vs.138.65 ± 6.38 ; Ⅲ layer:109.56 ± 8.69 vs.149.59 ±8.55;Ⅳ layer:l16.65 ±9.52 vs.155.76±9.87) (t=7.105,8.043,6.986,P＜0.05).Both CNTF positive cell number and gray scale in Ⅰ,Ⅴ,Ⅵ layers of visual cortex area 17 had no significant differences between two groups (P＞0.05).Conclusions Form deprivation in critical period of a new born animal may lead to distributing abnormality of CNTF in visual cortex,which maybe play a role in the development of form deprived amblyopia.

The ideal treatment and recovery of osteoarticular injury remain to be resolved. Small intestinal submucosa (SIS), a naturally-occurring decellularized extracellular matrix, has been recognized as an ideal scaffold for tissue engineering and widely used in repairing various tissues and organs. Nowadays its application has also been gradually increased in the field of orthopedics. We reviewed laboratorial studies and clinical trails about the application of SIS in bone and joint repair, aiming to evaluate its effects on the repair of bone, cartilage, meniscus, ligament and tendon. SIS has showed promising results in repairing bone, meniscus, ligament or tendon. However, additional studies will be required to further evaluate its effects on articular cartilage and tendon-bone healing. How to optimize SIS material,is also a focused problem concerned with making SIS a potential therapeutic option with high value for orthopedic tissue repair.
Animals ; Cell- and Tissue-Based Therapy ; Humans ; Intestinal Mucosa ; cytology ; Intestine, Small ; cytology ; Joint Diseases ; physiopathology ; surgery ; therapy ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds ; chemistry

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

Objective To investigate the genotype of metallo-?-lactamases (MBL) produced by carbapenem resistant Pseudomonas aeruginosa in pediatric patients.Methods 59 strains of resistance to imipenem or meropenem were collected from December 2003 to November 2005 in Beijing children's hospital.Isolates were further evaluated for MBL production by two screening methods.MBL Etest strips were used to screen the phenotype of MBL production.Molecular screening for blaVIM,blaIMP,blaSPM and blaGIM was carried out using primers targeting the conserved regions of the MBL genes.The PCR fragments obtained with integron primers were sequenced on both strands.The nucleotide sequences were compared with sequences available over the Internet.Results Of the 59 carbapenem resistant Pseudomonas aeruginosa included in this study,29 (49.2%)were MBL positive using Etest methods,and 39 (66.1%) of these tested positive for MBL genes by PCR.35 (89.7%) were positive for blaIMP genes and 4 (10.3%) were positive for blaVIM genes.All isolates were negative for SPM and GIM DNA sequencing revealed that the IMP-1 was detected in 35 IMP-producing isolates,and VIM-2 was detected in 4 VIM-producing isolates.Conclusions This study has demonstrated that MBL-producing strains in pediatric are more common than in adult.IMP-1-producing strains are the main in pediatric,and VIM-2-producing strains concurred.The production of MBL is one of the important reasons of carbapenem resistant Pseudomonas aeruginosa in pediatric.It is very important to monitor the production of MBL.

OBJECTIVETo investigate the pathogenesis of steroid-induced avascular necrosis of femoral head and the control mechanisms of Ground Beetle.

METHODSThe bone marrow-derived mesenchymal stem cells were cultured in vitro. The BMSCs in the 3rd generation were randomly divided into blank group, model group and Chinese medicine low, medium and high dose group. BMSCs of model group using high-dose steroid-induced in vitro were adipogenic differentiation to inhibit osteogenesis. Chinese medicine low, medium and high-dose groups at the same time were given the Ground Beetle intervention serum containing insects. The expression of osteocalcin, Alkaline phosphatase and Type I Collagen mRNA were detected and interfered 6 days later.

RESULTSSerum containing Ground Beetle could reverse content of steroid-induced alkaline phosphatase of BMSCs, the expression of ostercalcin and Type I Collagen mRNA decreased.

CONCLUSIONThe control mechanism of Ground Beetle on steroid-induced avascular necrosis is not only to improve the microcirculation, but also to inhibit steroid-induced osteogenic differentiation of BMSCs reduced.

Alkaline Phosphatase ; analysis ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Coleoptera ; Collagen Type I ; genetics ; Dexamethasone ; pharmacology ; Femur Head Necrosis ; chemically induced ; drug therapy ; Medicine, Chinese Traditional ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteocalcin ; analysis ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective:To observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers,such as alkaline phosphatase,Cbfαl and type I collagen,and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid induced ischemic necrosis of femoral head. Methods:Fifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs ,then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood,cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group ,model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro,and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week ,the expressions of alkaline phosphatase,Cbfαl and type I collagen mRNA were detected. Results:The alkaline phosphatase activity was signifi-cantly decreased in BMSCs of the model group as compared with the blank group (P<0.01),and the expression of Cbfαl and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly in-creased in BMSCs of each HSYA group as compared with the model group (P<0.05 or P<0.01),and the expression of Cbfαl and type I collagen mRNA were also increased significantly (P<0.05 or P<0.01). Conclusion:The mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

OBJECTIVETo study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneration and to explore the role of collagen types IX and X in disc degeneration.

METHODSFetal, adult and pathologic specimens were subjected to in situ hybridization with cDNA probes to investigate mRNA-expressions of types IX and X collagen gene.

RESULTSIn fetal intervertebral discs, positive mRNA hybridization signals of type IX collagen were concentrated in the nucleus pulposus and the inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus also showed positive type X collagen staining. Positive mRNA hybridization signals of types IX and X were not detected in the middle and outer layers of anulus fibrosus. In adult specimens, expression of type IX collagen mRNA was markedly decreased. No hybridization signals of type X collagen was observed. As for pathological specimens, there was no gene expression of type IX collagen. In severe degenerated discs from adults, there were focal positive expressions of type X collagen.

CONCLUSIONSObvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological discs. Results of type X collagen expression suggest that type X collagen is expressed only in older adult and senile discs (i.e., when disc degeneration has already reached a terminal stage), indicating the terminal stage of degeneration.

Adolescent ; Adult ; Collagen Type IX ; metabolism ; Collagen Type X ; metabolism ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Intervertebral Disc ; embryology ; metabolism ; Lumbar Vertebrae ; Male

Objective To investigate the effect of resveratrol on the expression of peroxisome proliferator activated re-ceptor γ coactivator-1α(PGC-1α)in skeletal muscle of rats with chronic obstructive pulmonary disease(COPD). Methods A total of 45 male Sprague Dawley rats were randomly divided into control group,model group and resveratrol group,15 rats in each group. The rats in the model group and resveratrol group were made COPD model through the intratracheal instillation of lipopolysaccharide and repeated smoke exposure,except the rats in the control group. From the 29th day of smoke exposure,the rats in the control group and model group were given 2 mL saline by gavage,once a day for 30 days;and the rats in the resvera-trol group were given 2 mL resveratrol solution by gavage(100 mg·kg - 1 ·d - 1 ),once a day for 30 days. After 30 days of con-tinuous gavage,the rats were sacrificed,then arterial blood and skeletal muscles were harvested. The level of tumor necrosis factor-α(TNF-α)in serum and skeletal muscle of rats was detected by enzyme linked immunosorbent assay. The expression of PGC-1α,nuclear respiratory factor 1(NRF1),mitochondrial transcription factor A(Tfam)and cytochrome C oxidase Ⅳ(COXⅣ)mRNA in skeletal muscle tissues of rats was determined by quantitative real-time polymerase chain reaction. The expres-sion of PGC-1α,NRF1,Tfam and COX Ⅳ protein was detected by Western blot. Results The level of TNF-α in serum and skeletal muscle tissues of rats in the model group and resveratrol group was significantly higher than that in the control group (P < 0. 01). The level of TNF-α in serum and skeletal muscle tissues of rats in the resveratrol group was significantly lower than that in the model group(P < 0. 01). The expression of PGC-1α,NRF1,Tfam,COX Ⅳ protein and mRNA in skeletal mus-cle tissues of rats in the resveratrol group and model group was significantly lower than that in the control group(P < 0. 01). The expression of PGC-1α,NRF1,Tfam,COX Ⅳ protein and mRNA in skeletal muscle tissues of rats in the resveratrol group was significantly higher than that in the model group(P < 0. 01,P < 0. 05). Conclusion Resveratrol can reduce the level of TNF-α in serum and skeletal muscle tissues of COPD rats,increase the expression of PGC-1α,thereby improving the mitochon-drial biosynthesis function.