In order to validate and estimate the capability of BMP9 to induce osteogenic differentiation of multipotent stem cells, three multipotent stem cells(C3H10, MEFs and BMSC) were used as target cells, and BMP9 was introduced into these cells by using recombinant adenoviruses assay, the effect of BMP9 on osteogenic differentiation of multipotent stem cells was demonstrated by using luciferase reporter assay, alkaline phosphatase(ALP) quantitative assay, calcium deposition assay, real time PCR, animal experiment and histological staining assay.The results demonstrated that BMP9 can induce ALP expression of C3H10, MEFs and BMSC by a dose dependent manner.BMP9 can also stimulate calcium deposition of C3H10 and MEFs in vitro, the osteogenic markers(ALP, Runx2, osteopontin, osteocalcin) were increased after stimulated by BMP9.BMP9 can activate canonical TGF?-Smad pathway, and promote the expression of osteogenic master gene Runx2.The animal experiment and histological staining assay show that BMP9 can induce ectopic bone formation in naked mice.To sum up, BMP9 is a more powerful cell factor to induce osteogenic differentiation of multipotent stem cells.

Objective To probe into the mechanisms of bone morphogenetic protein-2 (BMP-2) in osteosarcoma and to provide basis for gene therapy. Methods A 1.0 kb cDNA fragment of human BMP-2 gene was inserted reversely into PDOR and Human antisense BMP-2 retrovirus expression vector was constructed. The recombinant retroviral vector was transfected into human osteosarcoma cells OS-9901 with liposome AMINE that expressed abundant BMP. Positive cell clones were selected with G 418. The expression of BMP and proliferating cell nuclear antigen (PCNA) was determined by immunohistochemical ABC methods. The osteosarcoma cell cycle was analysed by flowcytometry, and the changes were observed by electronmicroscope. Results The expression of cellular BMP and PCNA in the transfected osteosarcoma cells decreased obviously (t=24.01， 26.09, respectively, P

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; epidemiology ; Prognosis ; Young Adult

OBJECTIVETo explore the expression patterns of the chemokine CXCL12 and its receptor CXCR4 in human sperm.

METHODSWe collected semen samples from 10 fertile men, performed density gradient centrifugation, and then determined the expressions of both CXCL12 and CXCR4 in the sperm by RT-PCR and immunofluorescence staining.

RESULTSRT-PCR revealed the mRNA expressions of CXCL12 (0.641 +/- 0.180) and CXCR4 (0.464 +/- 0.100) in the sperm. However, only CXCR4 rather than CXCL12 was expressed at the protein level, and the positive staining for CXCR4 was observed mainly in the posterior part of the acrosome.

CONCLUSIONCXCL12 and CXCR4 are involved as important molecules in regulating the function of human sperm.

Acrosome ; metabolism ; Centrifugation, Density Gradient ; Chemokine CXCL12 ; metabolism ; Humans ; Male ; Receptors, CXCR4 ; metabolism ; Signal Transduction ; Spermatozoa ; metabolism

Objective To evaluate the clinical effect of bandage contact lens on patients after the pterygium excision surgery.Methods 200 patients with pterygium excision were used retrospectively analysis and were divided into control group and observation group by random digital table method.Patients in observation group wore bandage contact lens while those in control group did not (traditional method).The degree of pain in both groups was evaluated at 8 hours,24 hours and 48 hours after surgery.In addition,patients were surveyed for the satisfaction of our treatment when discharge.Results The ocular foreign body sensation,eye burning,tears and eye open difficult were obviously decreased in observation group (t=5.643,10.315,P ＜ 0.01).The overall comfort level of the patients in observation group was obviously improved.pain feeling of the eye after surgery in 8h,24h and 48h markedly decreased in observation group (t=6.196-12.495,P ＜ 0.01).The degree of satisfaction scores of preoperative guidancein,postoperative pain control,emotion,mental nursing,self-care ability,comfort in observation group were (1.59-0.62) points,(1.83±0.70) points,(1.42±0.68) points,(1.42±0.68) points,(1.48±0.67) points,(1.29±0.77) points,(1.43-0.64)points which were obviously increased than (0.76±0.55) points,(0.83±0.64points,(0.82±0.56) points,(0.80±0.47) points,(0.79±0.61) points,(0.66±0.54) points (t=4.311-9.354,all P＜ 0.01).Conclusions Compared with traditional treatment,bandage contact lens can significantly reduce the discomforts after the pterygium excision surgery,and increase the postoperative comforts of patients.It proves that bandage contact lens have extensive clinical value.

Objective To make up an exercise prescription based on traditional Chinese medical training (EP-TCMT) for patients with stable chronic obstructive pulmonary disease (COPD).Methods Eighty-five pa- tients with stable COPD were randomly divided into a control group (CG group),a traditional Chinese medicine group ( TC group) and an exercise prescription group ( EP group).The patients in the TC and EP groups were giv- en intensive training for 8 weeks.Their 6 rain walk distance (6MWD) and Borg scale scores were assessed before and after the treatment.Results The 6MWD in the TC group increased from 337.68?59.18 m to 386.14?76.71 m,while those in the EP group improved from 348.00?55.94 m to 425.17?53.22 m.The Borg scale scores in the TC group decreased from 3.14?1.94 to 2.32?1.25,while those in the EP group declined from 3.45?1.84 to 1.72?0.70.Conclusion Making up EP-TCMTs is feasible.Additional treatment was found to improve exercise tolerance and decrease dyspnea in COPD patients.Exercise therapy based on traditional Chinese methods is easy and safe.

Objective To further define the molecular mechanism involved in the metastasis process in lung cancer and screen out the genes expressed differentially in the lung cancer.Methods mRNA differential display (DD-PCR) was employed to search the specific genes related to metastasis.The highly expressed fragments were cloned and sequenced.Compared with the data in GenBank,the homologous genes were found.The anchor primers were designed to validate the candidates from DD-PCR by real-time PCR.The structure of the gene was prognosticated by software.Results Nine differentially expressed genes were found.One of the nine genes,which named C3orf1,showed high different expression within the six tested cancer cells.The gene was 858bp long and encoded 285 amino acids.The molecular weight was about 32.2 kD.Analyzed by the bio-software,it was found that the gene was consisted with seven EGF-like domains (EGF_1),three 2Fe-2S ferredoxin/iron-sulfur binding regions (2FE2S_FER_1),two VWFC domains (VWFC_1),two thiolase active sites (thiolase_3),and so on.Conclusions The gene C3orf1 is over-expressed in the lung cancer 95D cells.It may encode a familiar secretary growth factor protein and play important role in stimulating growth of the cells.

Adult ; Anti-Inflammatory Agents ; therapeutic use ; Churg-Strauss Syndrome ; drug therapy ; pathology ; Follow-Up Studies ; Humans ; Male ; Methylprednisolone ; therapeutic use

[Summary] The genomic DNA was extracted from peripheral blood leukocyte of the patient with thyroid hormone resistance syndrome and 14 members of his family.The exons 1-10 of thyroid hormone receptor β (TRβ) gene were amplified by PCR.The products of PCR were sequenced directly to detect the gene mutation.The results showed that 3 members of this family were confirmed to have the C→T transition mutation at nucleotide 1 303 site within exon 10 of TRβ gene,and the missense mutation results in the substitution of histidine to tyrosine (H435Y).The heterozygous mutation may lead to the occurrence of thyroid hormone resistance syndrome.

Hepatitis C virus (HCV) genome is of high variation,which results in persistent infection of HCV and increases the incidence of liver cirrhosis and hepatocellular carcinoma.Following the successful paradigm established for HIV protease inhibitors,HCV NS3-4A serine protease has been selected as the main target for the development of small molecule antiviral agents.In this article,we review recent progress in the discovery and development of HCV NS3-4A protease inhibitors,and discuss their antiviral activities,pharmacokinetic properties,side effects and resistance profiles.