AIM: To construct lentiviral vector carrying the angiopoietin-1(Ang-1) gene,and make it express Ang-1 in the rat mesenchymal stem cells(rMSCs).METHODS: The cDNA encoding the CDS of Ang-1 gene was obtained from the placenta of the adult Fisher 344 rats with RT-PCR.After digestion with restrication endonuclease,the Ang-1 gene was recombined to construct the transfer plasmid PNL-Ang1-IRES2-EGFP.The three-plasmid system of lentiviral vector was consisted of PNL-Ang1-IRES2-EGFP,the packaging plasmid HELPER,and the envelope plasmid VSVG,which were co-transfected to 293T cells mediated by lipofectamin2000 to produce lentiviral particles.The rMSCs were infected by obtained lentiviral particles.The insertion of Ang-1 gene was detected by PCR,the mRNA expression of Ang-1 in rMSCs was detected with RT-PCR,the protein expression of Ang-1 was observed with immunocytochemistry and Western blotting methods.RESULTS: The result of sequencing showed that the cloned Ang-1 gene was consistent with the sequence reported in GenBank.After digestion with restrication endonuclease,the 1 512 bp fragment of Ang-1 gene and the 10.5 kb vector fragment of PNL-IRES2-EGFP were observed with gel electrophoresis.The insertion of Ang-1 gene in viral genome was confirmed.The EGFP expression was observed with the fluorescent microscope.In infected rMSCs,the mRNA and protein expressions of Ang-1 were confirmed.CONCLUSION: Lentiviral vector carrying Ang-1 gene has been successfully constructed.The infected rMSCs are able to express the Ang-1 mRNA and Ang-1 abundantly.This will facilitate the following exploratory development of Ang-1 gene-modified rMSCs.

Poly(ADP-ribose) polymerase-1 (PARP-1) has emerged as a promising anticancer drug target due to its key role in the DNA repair process. It can polymerize ADP-ribose units on its substrate proteins which are involved in the regulation of DNA repair. In this work, a novel series of para-substituted 1-benzyl-quinazoline-2, 4 (1H, 3H)-diones was designed and synthesized, and the inhibitory activities against PARP-1 of compounds 7a-7e, 8a-8f, 9a-9c and 10a-10c were evaluated. Of all the tested compounds, nine compounds displayed inhibitory activities with IC50 values ranging from 4.6 to 39.2 micromol x L(-1). In order to predict the binding modes of the potent molecules, molecular docking was performed using CDOCKER algorithm, and that will facilitate to further develop more potent PARP-1 inhibitors with a quinazolinedione scaffold.

PARP [poly(ADP-ribose)polymerase] represents a novel potential target in cancer therapy. It is involved in a DNA repair process by catalyzing the transfer of ADP-ribose units from NAD to a number of its substrate proteins. In this work, a series of novel azaindole derivatives was designed and synthesized. Moreover, 16 target molecules were screened and 8 compounds displayed inhibitory activity against PARP-1. It has been demonstrated that these azaindoles bearing cycloamine substituents at 2-position were active to both PARP-1 and PARP-2.

Poly(ADP-ribose)polymerase-1 (PARP-1) plays a significant role in the DNA repair process by catalyzing the transfer of ADP-ribose from NAD+ to its receptors. It is a promising anticancer drug target and many PARP-1 inhibitors have been developed and used in the clinical trial. In this work, a series of 3-(2-oxo-2-substituted acetamido)benzamides have been synthesized and their inhibitory activities against PARP-1 were evaluated. Of all the tested compounds, six compounds displayed inhibitory activities with IC50 values ranging from 0.23 to 5.78 µmol.L-1 . The binding pose of compound 5a was predicted using molecular docking to facilitate further structural modification.

ObjectiveTo compare the clinical effect of posterior circumferential fusion with versus without intertransverse process fusion on lumbar instability in aged patients. MethodsThe elderly with lumbar instability were treated with the posterior instrumented circumferential fusion technique in 80 cases. Among 58 patients followed up for at least 2 years, 28 cases (group A)underwent posterior circumferential fusion with intertransverse process, while 30 cases (group B)without intertransverse process. X-ray examination was used before and 1, 3, 6, 12 and 24 months after operation to evaluate the fusion condition of the bone graft, and visul analogue scale (VAS) and Oswestry disability index (ODI) questionnaire were applied to assess the pain of center back and leg,and the conventional activity.ResultsThe operation time and blood loss were more in group A[(185.3±56.6) min and (376.3±92.9) m1] than in group B [(146.4±46.3) min and (234.3±64.5)ml (t=12.37, 37.49, P＜0.01)]. All the 58 cases were followed for at least 2 years. The 27 cases (96.4％) of group A and 28 cases (93.3％) of group B got bone fusion (x2 =0.004, P＞0.05).There were marked differences in the VAS of center back at 1 and 3 months after operation between the 2 groups (t=3.178, 2.158, both P＜0.05), while no difference at the other 3 time point. VAS about the leg pain and the ODI showed no differences between group A and B after operation (all P＞0.05). ConclusionsFor the lumbar instability in the elderly, the posterior circumferential fusion with or without intertransverse process fusion can achieve a similar high rate of fusion and satisfactory clinical results,andtheposteriorcircumferentialfusionwithoutintertranaverseprocess is recommended for less trauma.

【Objective】To study the effect of light weight-bearing activity on postmenopausal osteoporosis.【Methods】36 female rats were randomly divided into 3 groups:① Sham,② Ovx (ovarietomized),③ Ovx+Im (ovarietomized and immobilized).All the group's maintained daily activity.And because of being immobilized,the right hind limbs of the third group lacked weight-bearing activity.12 weeks after ovarietomy,the BMD (bone mineral density),histomorphometry and biomechanics of the right femurs of rats were measured and analyzed.【Results】Comparing with the Sham group,the Ovx group's right femurs were manifested with the decrease of BMD,TBV (trabecular bone volume),MTT(mean trabecular thickness) and MCT(mean cortex thickness),while the increase of RS(resorption surface) and OS(osteotoid surface).Meanwhile their biomechanic nature declined.But statistically the BMD,MCT and the criteria of mechanical strength were not significant decrease.Otherwise,the Ovx+Im group's right femurs showed more apparent decrease of BMD,TBV,MTT and MCT.And the biomechanic nature was worse.Comparing with the Sham group,the BMD,MCT and the criteria of mechanical strength of the Ovx+Im group were statistically significant decreased.【Conclusion】If maintaining light weight-bearing activity,the ovarietomized rats were able to maintain relatively better bone quality.A lack of light weight-bearing activity wouldcause thedecline of bone quality.Thusthestudy suggested light weight-bearing activity was significantly effective on the prevention and treatment of postmenopausal osteoporosis.

0.05).Conclusions The local cell-mediated immunity of PTC is in the status of obviously suppressed.But the local impaired immune response against carcinoma is not associated with lymph node metastasis of PTC.

BACKGROUND: Epidural block(EB) is one of the effective and widely used remedies for lumbar disc herniation(LDH) . Animal experiments have been conducted to study the changes in the local tissue structure in the spinal canel after several treatments with EB.OBJECTIVE: To study the effect of EB for LDH on the local tissue structures in the spinal canel.DESIGN: A non-randomized and controlled clinical observation.PARTICIPANTS: Department of Pathology and Laboratory of Electron Microscope of an Affiliated Hospital of a University.SUBJECTS: Forty patients with LDP, who failed to respond to EB treatment and subsequently received discectomy in the Department of Orthopaedics,First Affiliated Hospital of Sun Yat-sen University from May 1992 to December 1997, were enrolled in this study, with 20 patients receiving discectomy but not EB treatment serving as the control group.METHODS: From the 60 cases, totally 120 specimens of yellow ligament and the disc tissues were obtained during discectomy for optical microscopical examination. A portion of the yellow ligament and the lumbar disc tissues were chosen from EB group(12 cases) and non-EB group(10 cases), respectively, for electron microscope examination(involving altogether 44specimens) . The changes in histology and ultrastructnres in the specimens were compared between the two groups.MAIN OUTCOME MEASURES: ① Main results: the reactive proliferation of the tunica adventitia and inflammatory reaction of the intervertebral disc and yellow ligament; ② Secondary results: changes in the ultrastructure of the anulus fibrosus, nucleus pulposus and yellow ligament.RESULTS: No significant difference was found between the two groups either in light microscopy or electron microscopy in the degree of smoothness and the inflammatory reaction of the cavosurface of the yellow ligament and the disc tissues, nor in the cellular ultrastructures and fibrous arrangement.CONCLUSION: There is no direct relationship between lumbar EB and epidural scar adhesion formation. EB does not affect the ultrastructures of the local tissues in the spinal canal, but care must be taken of the sterilization in the surgical procedure, as chronic inflammatory reaction in the yellow ligament and disc tissues can be present in some patients.

Aim To observe the infarct size, apoptosis of the neurocytes,the expressions of Bcl-2 and Bax pro-teins after focal cerebral ischemia-reperfusion injury (CIRI) in the brain tissue of rats and the protective effects of rofecoxib.Methods The model of local CIRI was induced by reversible middle cerebral artery occlusion (MCAO) with inserting a thread through internal carotid artery,2 h occlusion followed by 24 h reperfusion.Rofecoxib was administrated (ig) at the reperfusion initiation. Using TTC staining technique to measure the infarct size of brain,TUNEL technique to examine apoptosis of the neurocytes,immunohistochemical method to examine the expression level of Bcl-2 and Bax proteins in brain tissue.Results After focal CIRI,the infarct focus of brain was showed,both apoptosis rate of the neurocytes and Bax protein expression were significantly increased and the ratio of Bcl-2 to Bax was significantly reduced.With the use of both 1.12 and 2.24 mg?kg -1 doses of rofecoxib,the brain infarct size was dramatically reduced. With the use of 2.24 mg?kg -1 dose of rofecoxib,both apoptosis rate of the neurocytes and Bax protein expression were significantly decreased,both Bcl-2 protein expression and the ratio of Bcl-2 to Bax were significantly higher than that in the group Model.Conclusion The highly selective COX-2 inhibitor rofecoxib may increase Bcl-2 protein expression and decrease Bax protein expression then increase the ratio of Bcl-2 to Bax and so reduce the neurocytes apoptosis in brain tissue thus significantly improve the brain injury after CIRI.

Objective To investigate the potential association between SEPS1 variants and Hashimoto's thyroiditis (HT)in the Chinese Han population and evaulate the effects of SEPS1 haplotypes on the susceptibility to immunity-mediated diseases.Methods We analyzed 7 SNPs with minor allele frequency (MAF)≥0.05 in a cohort of 1013 HT patients and 2998 healthy controls from genetically independent Han Chinese individuals.Single SNP,haplotype and gender-specific associations were performed. Results We found that rs28665122 was significantly associated with HT,and SNP was associated with HT in females.Further haplotypic analysis indicated that a haplotype block (rs2009895-rs28665122)spanning the promoter of SEPS1 gene was highly associated with HT. Additionally,the haplotype CA in the block had an almost 1.5-fold increase in these cases,which remained significantly associated with HT in females.Conclusion SEPS1 plays a role in the pathogenesis and etiology of HT.