Objective:To investigate the effects of the treatment of early cracked teeth with vital pulp preservation.Methods:63 early cracked teeth with vital pulp in 56 patients were treated by light cured composite resin filling with or without MTA rebasing,and restored by precious metal crown.The patients were followed up for 2 years.Results:After 2 years follow-up,57 teeth were successfully treated and failure was found in 6 teeth,the effective rate was 90.4％.Conclusion:Light cured resin filling directly or with MTA rebasing for early cracked teeth with vital pulp,and restoration by precious metal crown are effective in the treatment of early cracked teeth with vital pulp.

Objective To study the changes of microarchitecture, bone mineral density (BMD) , and bone mineral content (BMC) in apolipoprtein E knockout( ApoE-/-) mice. Methods Male ApoE-/- mice at 15, 28, and 40-week of age and sex-age-matched wild-type (WT) mice were involved in the study. The trabecular and cortical bone microarchitecture were assessed by micro-CT( μCT) in the right distal femur. The total body BMD of the left femur was determined by dual-energy X-ray absorptiometry ( DXA). The relationships among BMD, microarchitecture, and BMC were analyzed. Results Compared with WT mice,the advancing age ApoE-/- mice showed an increased volumetric BMD (vBMD), tissue BMD (tBMD) , BMC, bone volume fraction (BV/TV), trabecular number (Tb. N ) , trabecular thickness (Tb. Th) with an decreased bone surface fraction ( BS/BV), trabecular separation (Tb. SP) , and the structure mode index (P <0. 05 ) in the cancellous bone of femur. The cortical bone microarchitecture parameters as inner perimeter, outer perimeter, cortical area, marrow area, total area and moment of inertia were also increased, but cortical BMD, cortical bone mineral content (C. BMC) and cortical thickness retained constant. At the age of 28 weeks,the total body BMD in ApoEE-/- mice revealed higher than WT mice (P<0. 05) and there was no changes in 15 and 40-week-old mice compared with the sex-age-matched controls. vBMD was positively correlated with BMC, BV/TV,Tb. Th, BS/BV, and C. BMC, with the correlation coefficients 0.955,0.944,0. 834,0.923, and 0.903 .respectively, and there was no correlation between vBMD and the other parameters. Conclusions ApoEE-/- mice display an increased bone mass, suggesting that ApoE has an important role in bone remodeling.

OBJECTIVE:To observe clinical efficacy and safety of individual antiviral therapy of tenofovir combined with inter-feron α1b for chronic hepatitis B (CHB). METHODS:96 CHB patients were randomly divided into control group,observation group A and observation group B,with 32 cases in each group. Control group was given entecavir orally,0.5 mg,qd;observation group A was given tenofovir orally,1 piece,qd;observation group B was additionally given interferon α1b,50 μg,3 times a week,on the basis of observation group A. The treatment course lasted for 48 weeks in 3 groups. Clinical efficacy of 3 groups was compared,and the changes of serum liver function indexes,HBV-DNA negative conversion rate and the occurrence of ADR were compared before and after treatment. RESULTS:The total effective rate of observation group B(84.38%)was significantly higher than that of observation group A(62.60%)and control group(37.50%),and that of observation group A was significantly higher than control group,with statistical significance (P<0.05). Compared with before treatment,the serum levels of AST,ALT and TBIL were significantly decreased after treatment in 3 groups;the observation group B were significantly lower than those of obser-vation group A and control group,with statistical significance(P<0.05);there was no statistical significance between observation group A and control group(P>0.05). The negative rate of HBV-DNA in observation group B were significantly higher than those in control group and observation group A after 12 and 24 weeks of treatment,with statistical significance(P<0.05);there was no statistical significance between observation group A and control group (P>0.05). No obvious ADR was found in 3 groups. CON-CLUSIONS:Tenofovir combined with interferon α1b shows significant clinical efficacy for CHB,and is significantly better than that of entecavir and tenofovir alone.

Traditional Chinese medicine (TCM) cultural tourism, with the trait of health and unique features, is based on rich resources of medicine and profound TCM culture. TCM cultural tourism focuses on making tourists get TCM knowledge and enjoy TCM tourism activities. Mawangdui Han Dynasty Tomb, a key scenic spot to develop TCM cultural tourism in Changsha, has advantages of high TCM cultural value with a good geographical position, and superior geographic and tourist viewing conditions. It has a great value in TCM cultural tourism.

Dendrobii Officinalis Caulis is considered as one kind of rare and precious Chinese medicinal herb with the efficacy of nourishing yin fluid, enhancing physique, benefiting and tonifying spleen and stomach, protecting liver and cholagogue, strengthening gluten, lowering lipid and blood sugar, inhibiting tumor, brightening eyes, nourishing skin, and prolonging life. This article used SWOT method to analyze the strength, weakness, opportunity, and threat of Dendrobii Officinalis Caulis respectively so as to analyze its marketing prospects in Hunan Province comprehensively. On the basis of the analysis, the early stage and late stage strategies of development of Dendrobii Officinalis Caulis in Hunan Province was carried out, which played a role in its medicinal value, economic value, and social value.

OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells),and to learn more about the nuclear responses to mitochondrial dysfunction.METHODS The nuclear proteomes of Rho and Rho + cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies,the differentially expressed protein- spots were identified by MALDI-TOF mass spectrum (MS),the nucleophosmin and P53 expression were detected by Western blotting assay,and the mitochondrial memhrane potential (MMP) was measured by the laser scanning confocal microscope.RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei.SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei.One down-regulated protein-spot was identified as elF-6,and 4 up-regulated proteinspots were identified as nucleophosmin,SFRS1,SFRS3 and hnRNP G,respectively.The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting.Based on the clues from proteomic analysis,P53 expression in Rho0 cells was higher than in Rho + cells,and MMP was consistent in Rho + and Rho0 cells.CONCLUSION mtDNA-depletion induces nuclear proteome alteration.Rho0 cells can be used as a model to study the crosstalk between mitochondrion and nucleus.

AIM:Toinvestigatetheeffectsofcalcitoningene-relatedpeptide(CGRP)onmyocardinexpres-sion and phenotypic switch in vascular smooth muscle cells ( VSMCs) .METHODS:VSMCs were obtained by aortic tissue adherent culture and treated with angiotensin Ⅱ( AngⅡ) , AngⅡ+CGRP or AngⅡ+CGRP +CGRP8-37 .The protein expression of myocardin and the phenotypic proteins of the VSMCs was detected by Western blot .RESULTS:The expres-sion of myocardin in cultured VSMCs showed downregulation along with time expansion .The protein level of myocardin was higher at 48 h and 72 h than that at baseline in the cultured VSMCs (P<0.05).However, the myocardin was lower at 48 h and 72 h than that at baseline after treatment with CGRP in cultured VSMCs (P<0.05).Furthermore, at 48 h in cul-tured VSMCs, the myocardin decreased along with α-smooth muscle actin (α-SMA) (P<0.05), and osteopontin (OPN) increased (P<0.05) in AngⅡ group compared with control group .After treatment with CGRP, the levels of myocardin andα-SMA become higher ( P <0.05 ) but OPN was lower ( P <0.05 ) in CGRP group than those in AngⅡ group. CGRP8-37 abrogated CGRP-induced increase in myocardin and α-SMA and decrease in OPN in CGRP 8-37 group compared with CGRP group .CONCLUSION: CGRP may regulate the phenotypic switch of the VSMCs and maintain the cells in contractile phenotype through the upregulation of myocardin protein , which may be accomplished by the combination of CGRP and its receptor .

Objective To explore the effect of transplantation of mesenchymal stem cells (MSCs) transfected with the human receptor activity modifying protein 1 (hRAMP1) gene on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) after carotid balloon angioplasty was performed in rabbits with carotid atherosclerosis.Methods Density gradient centrifugation and adherent culture were carried out to obtain MSCs,which were then transinfected with an adenovirus vector carrying the hRAMP1 gene or an empty adenovirus vector.A rabbit model of atherosclerotic stenosis and balloon angioplasty was successfully established.Results were randomly divided into three groups:the hRAMP1-MSCs group,theadipose-derived MSCs (Ad-MSCs) group and the control group.MSCs were transinfected with Ad-EGFP-hRAMP1,Ad-EGFP or PBS by transplantation into the injured carotid arteries.Homing and differentiation were assessed with MSCs harvested at 7 d.With MSCs collected at 28 d,Western blotting was used to measure the expression of the hRAMP1 target gene in the carotid artery; the neointima and media area in the injured carotid arteries were estimated; carotid artery morphology was examined with H&E staining; and the proliferation and apoptosis of VSMCs were determined by immunohistochemistry and TUNEL.Results The expression of CD31 and EGFP was found in proliferating neointima lesions at 7d in the hRAMP1-MSCs group and the Ad-MSCs group.At 28d of MSC transplantation,the level of RAMP1 significantly increased in the hRAMP1-MSCs group,compared with the Ad-MSCs and control groups [(63.0±4.9) vs.(28.3±2.5) and (27.2±7.2),all P＜0.05],but there was no differencein the RAMP1 level between the Ad MSCs group and the control group (P＞0.05).Positive expression of the α-smooth muscle antibody (α-SMA) was found in all three groups at 28 d of MSC transplantation.The thickness of the hyperplastic neointima significantly decreased in the hRAMP1-MSCs group,compared with the other two groups (P＜0.05),and was lower in the Ad-MSCs group than in the control group (P＜0.05).The expression of proliferating cell nuclear antigen (PCNA) was lower in the hRAMP1-MSCs group than in the Ad-MSCs and control groups at 28d of MSC transplantation (P ＜0.05),while the PCNA level was lower in the Ad-MSCs group than in the control group (P＜ 0.05).The VSMC apoptosis rate significantly increased in the hRAMP1-MSCs group,compared with the Ad MSCs and control groups (P＜0.05),and was the lowest in the control group (P＜0.05).Conclusions Gene-modified stem cell therapy can effectively inhibit vascular intimal hyperplasia,thereby reducing restenosis after angioplasty.

Objective To observe the effect of adenovirus- receptor activity modifying protein-1 (RAMP1) on nuclear factor(NF κB) and myocardial fibrosis in heart of rabbit with myocardial infarction. Methods Myocardial infarction (MI) models were developed in 54 rabbits and they were randomly divided into RAMP1 group,EGFP group and control group according to whether pAd2-EGFP-RAMP1,pAd2-EGFP or PBS was injected into infarction region and its border.At 7 d,14 d and 28 d after injection,Left ventricular function indices such as LVEF,LVSd,LVDd and LVFS were estimated by echocardiogram,the expression of NF-κB in myocardium was detected by Western blot,the plasma level of TNF-α was measured by ELISA,and fibrosis and collagen content was measured by Masson stain. Results At 7 d after adenovirus injection,the expression of RAMP1was significantly increased in RAMP1 group (67.33 ± 3.97)％ as compared to EGFP group(20.59 ±3.26) ％ and PBS group ( 23.80 ± 5.08) ％ ( P ＜ 0.05 ).The expression of NF κB was decreased in RAMP1 group ( 26.54 ± 5.13 ) ％ versus EGFP group (62.60 ± 6.18) ％ and PBS group (62.95 ±5.17)％ (P＜0.05).The plasma level of TNF-α was lower in RAMP1 group than in EGFP group and in PBS group at different time[7 d:( 136.74 ± 5.42) μg/L vs.( 196.97 ± 14.17) μg/L,(203.67 ±13.90)μg/L; 14 d:( 154.51 ± 13.61 )]μg/L vs.( 112.22±6.74 )μg/L,(160.46± 14.27)μg/L ;28 d;(51.10± 5.62)μg/L vs.( 95.55 ± 9.94 )μg/L,( 98.96 ± 12.68) μg/L,all P＜ 0.05].The collagen content was reduced in RAMP1 group as compared with EGFP group and PBS group at 14 d and 28 d [14 d:(7.10±0.98)％ vs.(19.52±2.32)％,(17.91±0.96)％ ;28 d:(17.04±2.44)vs,(34.10±5.59) ％,(33.98±4.33)％,all P＜0.01].At 28 d after infarction,the infarct size was decreased in RAMP1 group (26.54 ± 5.13) ％ compared with EGFP group (32.20 ± 3.73) ％ and control group (35.58±2.65) ％ (P＜0.01),and better heart function appeared in RAMP1 group. Conclusions The high-expression of RAMP1 could decrease the collagen deposition and fibrosis in the border of infarction and improve heart function through lower expression of NF-κB and decreasing the plasma level of TNF-α.

Objective: To explored the effect and related mechanism of miRNA-21 on hydrogen peroxide-induced C-kit⁺ cardiac stem cells apoptosis. Methods: C-kit⁺ cardiac stem cells were isolated from SD rats by the methods of enzyme digestion and magnetic bead. Cells were divided into the following experimental groups: (1) negative control mimics (NCM)group: cells were transfected with negative control miRNA-21 mimics for 48 hours; (2)mimics group: cells were transfected with miRNA-21 mimics for 48 hours; (3) NCM+ H₂O₂ group: negative control miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H₂O₂ for 2 hours; (4)mimics+ H₂O₂ group: miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H₂O₂ for 2 hours. mRNA of miRNA-21 was detected by RT-PCR. The apoptosis rate of C-kit⁺ cardiac stem cells was determined using the annexin V-FITC/PI staining assay. Western blot was employed to measure the expression of apoptosis related proteins(Caspase-3, Bax, and Bcl-2). Results: (1) Compared with the NCM group, the mRNA expression level of miRNA-21 was significantly up-regulated in mimics group, while obviously down-regulated in NCM+ H₂O₂ group(all P<0.05). Compared with the mimics group, the mRNA expression levels of miRNA-21 in mimics+ H₂O₂ group was significantly downregulated (P<0.05), but remarkably upregulated compared with the NCM+ H₂O₂ group(P<0.05). (2) Flow cytometry results indicated that the early apoptosis rates were similar between the NCM group and mimics group ((4.57±3.45)% vs. (5.13±3.21)%, P>0.05). Compared with the NCM group, the early apoptosis rates were remarkably increased ((79.07±5.75)% vs.(4.57±3.45)%, P<0.05) in NCM+ H₂O₂ group. Compared with the mimics group, the early apoptosis was significantly up-regulated in the mimics+ H₂O₂ group ((30.27±1.36)% vs.(5.13±3.21)%, P<0.05), which were further down-regulated in mimics+ H₂O₂ group compared with the NCM+ H₂O₂ group ((30.27±1.36)% vs.(79.07±5.75)%, P<0.05). (3) Western blot results showed similar protein expression of Caspase-3, Bax and Bcl-2 between NCM group and mimics group(all P>0.05). Compared with the NCM group, the Caspase-3 and Bax protein expression was significantly increased in NCM+ H₂O₂ group (all P<0.05), but the protein expression level of Bcl-2 was similar between the 2 groups(P>0.05). The Caspase-3 and Bax protein expression was markedly decreased, while Bcl-2 apparently increased in the mimics+ H₂O₂ group compared with the NCM+ H₂O₂ group(all P<0.05). Conclusion Overexpression of miRNA-21 protects the C-kit⁺ cardiac stem cells from apoptosis caused by oxidative stress through downregulating proapoptotic and upregulating the antiapoptotic proteins.