OBJECTlVE To investigate the correIation between baicaIin metaI(Ni2+,Co2+,Cu2+) compIexes(BmC)with their anti-tumor activity and the abiIity of BmC to bind to hepatoma ceII DNA. METHODS The cheIating Iigand method was used to synthesize BmC,and the composition and struc-ture of BmC were characterized. mTT,PI staining method and AnnexinⅤ-FITC doubIe staining method were used to anaIyze the effect of BmC on SmmC-7721 ceII proIiferation,cycIe and apoptosis,and to expIore their cytotoxic effect on SmmC-7721 ceIIs in combination with morphoIogy. With DNA extracted from hepatoma ceIIs as a target,cycIic voItammetry and AC impedance were used to study the interaction of BmC with DNA. The interaction mechanism between BmC and DNA was expIored. RESULTS Three new types of BmS were successfuIIy prepared. The moIecuIar formuIas of compIexes were Na2 Ni(C21 H16 O11 )2·10H2 O,Na2 Co(C21 H16 O11 )2·8H2 O,and Na2 Cu(C21 H16 O11 )2·8H2 O,respec-tiveIy. CeII proIiferation and morphoIogy detection reveaIed that BmC 6.25-100 mg·L-1 treatment for 24, 48 and 72 h couId inhibit SmmC-7721 ceII survivaI. BmC cytotoxicity was Iisted as foIIows:baicaIin-cop-per( BC-Cu)﹥ baicaIin-cobaIt( BC-Co)﹥ baicaIin-nickeI( BC-Ni)﹥ baicaIin( BC),in a concentration-dependent manner(P﹤0.01)and time-dependent manner(P﹤0.01). According to the resuIts of ceII cycIe and apoptosis detection,BmC retarded the growth of ceIIs from G0 / G1 phase into S phase or G2 / m phase whiIe inducing apoptosis of SmmC-7721 ceIIs. The resuIts of eIectrochemicaI anaIysis showed that BmC and hepatoma SmmC-7721 ceII DNA formed a non-eIectroactive supermoIecuIar compound through the mixed-mode of eIectrostatic interaction and insertion effect. The binding parameters were obtained:the binding number m = 2,the binding constant βBC = 2.77 ×106 L·moI-1 ,βBC-Ni = 5.46 ×106 L·moI-1 ,βBC-Co =7.74×106 L·moI-1 ,and βBC-Cu =1.21×107 L·moI-1 . The abiIity of BmC to bind to DNA was signifi-cantIy enhanced by BC compIexes with metaI ions,and their abiIity was Iisted as foIIows:BC-Cu﹥BC-Co﹥BC-Ni﹥BC. CONCLUSlON BmC shows cytotoxicity by bIocking the ceII cycIe,inhibiting ceII proIiferation and motivaing apoptosis. The abiIity of BmC to bind to DNA is consistent with its cytotoxicity. BmC,after binding to ceII DNA,wiII bIock DNA repIication,inhibit ceII proIiferation,Iead to ceII apoptosis,and exhibit anti-tumor activity.

The interaction mechanism between rhaponticin (RT) and human serum albumin (HSA) has been studied by fluorescence spectroscopy and absorbance spectra. The mediation effect that the metal ions took part in the interaction has also been discussed in this paper. Based on different theoretical models of fluorescence quenching, the binding constant (K) and binding sites (n) of the interaction were determined and analyzed comparatively. The quenching mechanism of the binding reaction has also been discussed. The binding distance (r) and energy-transfer efficiency (E) between RT/RT-Co(II)/RT-Ni(II) and HSA were also obtained by virtue of the Förster theory of non-radiation energy transfer. The effect of RT acting on the HSA's conformation was analyzed by synchronous fluorescence spectroscopy. The result showed that the result calculated by different theoretical models is generally equivalent and RT bound HSA strongly by forming stable complex, which indicates that HSA under physiological conditions can act as a carrier for RT to be transported to exert effects. The microconformation of HSA changed significantly due to hydrophobicity change in the chemical environment of some fluorescence chromophores in the subdomain IIA and IIB of HSA. Metal ions Co(II) and Ni(II) can mediate RT-HSA interaction, making the binding of the drug to protein stronger, which indicates that Co(II) and Ni(II) can enhance rhaponticin's medical efficacy under physiological conditions.

Aged ; Amyloidosis ; therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Moxibustion ; methods ; Skin Diseases ; therapy

Chelating ligand method has been used to synthesize baicalin-metal (Ni2+, Co2+, Cu2+) complexes (BMC). The composition and structure of BMC were characterized by the element analysis, ultraviolet spectrum (UV), infrared spectrum (IR), mass (MS) and thermal gravitational analysis (TGA). MTT was used to analyze the effects of BMC on SMMC-7721 cell proliferation. PI staining method and Annexin-V/FITC double staining method were used to analyze the effects of BMC on the cell cycle and apoptosis of SMMC-7721 cell. Fluorescence quantitative RT-PCR was used to analyze the expression of BMC on Bcl-2 gene and Bax mRNA, flow cytometry was used to analyze BMC on the expression of Bcl-2 protein and Bax protein. The antineoplastic activity and mechanism of action of BMC was explored comprehensively. The results showed that three new kinds of BMC (molar ratio of 2 : 1) were successfully prepared, the complexes molecular formula are: Na2Ni(C21H16O11)2 x 10H2O, Na2Co(C21H16O11)2 x 8H2O and Na2Cu(C21H16O11)2 x 8H2O. According to the results of cell cycle and apoptosis detection, BMC stopped cells at G0/G1 phase to S phase and G2/M phase. Gene and protein detection showed that under the given concentration and time, BMC can downregulate the expression of Bcl-2 gene in SMMC-7721 cells, and significantly decrease the expression of Bcl-2 protein, at the same time, with the increase of expression of Bax gene, the Bax protein's expression increased significantly. Which indicates that BMC restrain cell proliferation and cell apoptosis by stopping cell cycle, reducing the expression of Bcl-2 and increasing that of Bax; The anti-tumor activities of three kinds of complexes were: baicalin-copper (BC-Cu) > baicalin-cobalt (BC-Co) > baicalin-nickel (BC-Ni) > baicalin (BC), showing the dose-response relationship.

Objective To evaluate the treament of central retinal artery occlusion by thrombolysis infusion via super selective ophthalmic artery catheterization.Design Retrospective,observational case series.Participants 21 eyes of 21 patients with CRAO. Methods 21 patients with CRAO were diagnosed by stereoscopic color fundus photography and flouorescein fundus angiography, and were treated by urokinase infusion via super select ophthalmic artery catheterization seldinger technique.Main Outcome Measures Visual acuity and the postoperative complications.Results In the 21 patients,10 had showed the occlusion of ophthalmic arterial trunk by super selective internal carotid artery angiography,the others can be found the appearance of ophthalmic arterial trunk and all patients had undergone thrombolysis therapy successfully.Imaging times of central retinal artery before and after thrombolysis infusion treatment are 38.18?10.86 seconds,12.65?3.30 seconds(t=-11.89,P=0.000).Mean foflow-up time is 3.23?1.26 months.After the treatment,the visual acuity was more than 0.25 in 4 patients,improved to different extent in 9 and remained unchanged in 8. Conclusions Super selective arterial catheterization with thrombolysis for CRAO can improve the visual acuity of the patients,a speedy execution of all internal,neurological,and ophthalmology diagnostic measures;and a prompt therapy are necessary.

Objective To value the clinical application of diffusion tensor imaging (DTI) in cerebral diseases. Methods Six volunteers and 6 patients (including 3 patients with ischemic stroke and 3 patients with glioma) were examined by DTI and T1weighted, T2weighted MR scan. All data were processed with DtiStudio software to show the white matter fiber tracts. The fractional anisotropy (FA) of the diffusion tensor were measured between the affected and the unaffected side. Results The white matter fiber tract could be observed clearly on the FA map. The pyramidal tract with different degree disruption could be showed in 3 patients with ischemic stroke. Compression, displacement, infiltration or destruction of pyramidal tract, corpus callosum or internal capsule and external capsule could be seen in 3 patients with glioma, and FA was significantly reduced on the affected side as compared to the unaffected side. Conclusion Diffusion tensor imaging is useful in observing the damage and displacement of the white matter fiber tracts in vivo, beneficial to the surgical plan for patients and prognosing recovery of function.

Animals ; Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Hydrogen-Ion Concentration ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Oxygen ; metabolism ; Rats

Objective To investigate the roles of adrenomedullin(ADM) and endothelin-1(ET-1) in left-to-right shunt congenital heart disease(CHD) complicated with congestive heart failure(CHF).Methods Plasma ADM and ET-1 concentrations were measured by radioimmunoassay in heart failure stage and recovery period of 18 infants with left-to-right shunt CHD.Also 20 healthy infants acted as control group were measured.Results Plasma concentrations of ADM and EF-1 were significantly higher in heart failure stage of left-to-right CHD infants than those in their recovery period,and both in their heart failure stage and in their recovery period were higher than that in control group(P

Objective To explore the effects of iPS cells-derived chimeric thymus transplantation on T cells reconstitution and graft versus host disease of murine after allo-BMT. Methods iPS cells-derived chimeric thymus was grafted under the renal capsules of mice after allogeneic IBM-BMT. The mice were divided into three groups:IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group. Four weeks after BMT, T lymphocyte subsets in the peripheral blood were analyzed by flow cytometry, the degree and pathological examination of GVHD were observed, respectively. Results Percentage of CD8+T cells in IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group was(5.52 ± 0.83)%,(11.10 ± 1.49)%and(8.49 ± 0.82)%respectively, there was signifi-cant difference between pairwise comparisons(P<0.05), and percentage of CD4 + T cells of the peripheral blood in IBM-BMT+TT group(9.60 ± 0.69)%was significantly higher than IBM-BMT group(6.42 ± 1.40)%and IBM-BMT+DLI group(8.07 ± 0.65)%(P<0.05) . IBM-BMT group and IBM-BMT+TT group showed less clinical and histopathological scoring of GVHD than IBM-BMT + DLI group. Conclusion iPS cells-derived chimeric thymus transplantation could effectively accelerate T cells reconstitution and prevent GVHD after allo-BMT.

Objective To examine an in vivo method for the differentiation of induced pluripotent stem cells (iPSCs) into thymic epithelial cells (TECs) in mice. Methods Green fluorescent protein-expressing iPS cells, derived from C57BL/6 mice, were injected into blastocysts from ICR mice. Chimeric blastocysts were then transferred into uteri of E2.5 pseudopregnant mice. Chimeric mouse could be identified by coat color 10 days after birth. The chimeric thymus was transplanted under the renal capsule of BALB/c nude mice. The spleen was cut out from the thymus-transplanted nude mice and the cells were dispersed and analyzed by a flow cytometer 4 weeks after transplantation. Results Chimeras were born 17 days after embryo transfer and 13 live-born chimeras were obtained. The contribution of iPSC-derived cells in the chimeras ranged from 5% to at most 90%. Typical thymic epithelium structure consisted of green fluorescent protein-expressing cells in chimera. The iPSCs-derived thymic epithelial cells could support the generation of new T cells. Conclusion The results indicate that mouse iPS cells can differentiate in vivo towards normally functioning TECs.