Objective To explore the effects of fluoride with different doses on the expressions of TGF-? and Smad2/3 in fibroblast and osteoblast at different periods.Methods Fibroblasts and osteoblasts were exposed to different concentrations of fluoride(0,0.0001,0.001,0.1,1,10 and 20 mg?L~(-1)).The levels of TGF-? protein and Smad2/3 at 2,4,24,48 and 72 h after treatment were measured by using ELISA method.The expression of TGF-? mRNA was tested with RT-PCR method.Results In fibroblasts,the contents of TGF-? protein were decreased in the groups of 0.001,0.1,1,10 and 20 mg?L~(-1) F~-at the time of 2 h and in the groups of 1.0001,0.001,0.1,10 and 20 mg?L~(-1) at the time of 4 h(P

The morphology and position of the AV node and AV bundle were observed in 13 human hearts with serial sections. 1.the AV node is a long sagittal flatt ened structure, its transverse section is triangular in shape with a right convex surface, sometimes the cross section is fusiform or half oval in shape. Its size is 3.5x3.3x1.1 mm in adult. In 5 cases the endocardium lying on the right surface of the AV node is elevated.2.The AV node is situated in the upper border of the atrioventricular septum (between the levels of the attachment lines of the mitral and tricuspid valves). The adult AV node is 1.8-5.8 mm anterior to the coronary sinus orifice, 0.3-0.7 mm from the endocardium of the right atrium, 3.3-7.5 mm above the upper border of the septal leaflet of the tricuspid valve. The left surface of the AV node contacts with the central fibrous body.3.The AV node can be divided in 2 parts: superficial and deep, the fibers of the super ficial part are longitudinal in sections and end in the lower border of the AV node. In one case, the deep part is subdivided in an upper part and a lower part. In the specimens in which the right atrial endocardium lying on the right surface of the AV node is elevated, the overlaying fibers end in the endocardium. At the upper border, right surface, and posterior margin of the AV node, there are atrial fibers ending to the AV node. 4. The adult AV bundle is 5.7-7.9 mm long, 1.1-1.5 mm in diameter. Its anterior part is on top of the muscular interventricular septum in 7 specimens, on its left surface in 3 specimens, and in the substance of the muscular interventricular septum in 2 specimens. In one case its course is very special, at first on the top of muscular interventricular septum, then at its left surface, finally in the substance of the right part of the muscular interventricular septum.

Objective To explore the therapeutic effect of phenylethanoid glycosides on cyclophosphamide (CTX)-induced dyszoospermia in mice and to preliminary elucidate the mechanisms involved in the process. Methods Phenylethanoid glycoside was extracted by ethanol extraction.Forty male BALB/C mice were randomly divided into control group,model group,low dose of phenylethanoid glycosides group (50 mg· kg-1 )and high dose of phenylethanoid glycosides group (100 mg·kg-1 ).Except control group,the dyszoospermia mouse model was established by peritoneal injection of CTX at the daily dose of 80 mg· kg-1 ,once daily for successive 5 d. After modeling, phenylethanoid glycosides were intragastrically administered at corresponding doses to each phenylethanoid glycosides group.Equal volume of normal saline was given to the mice in control group and model group by gastrogavage.All the medication was performed once daily for successive 30 d.The testis tissue was obtained 24 h after the last intragastric administration.The level of testosterone in the testis tissue homogenate was determined by enzyme linked immunosorbent assay.The sperm counts, the motility rates, and the teratospermia rates in various groups were compared.The morphological changes of the testis tissue were observed using HE staining.Results Compared with control group, the sperm count and the motility rate were decreased, the teratospermia rate was increased,and the testosterone level in the testis tissue homogenate was decreased in model group(P<0.01).Compared with model group,the sperm counts and the motility rates were increased,the teratospermia rates were decreased, and the testosterore levels in the testis tissue homogenate were increased in phenylethanoid glycosides groups (P<0.05 or P<0.01).The histological results showed atrophy and degeneration of seminiferous tuble,thicker seminiferous epithelium and azoospermic lumina in model group;the number of seminiferous epithelial layers was increased and the seminiferous cells orderly arranged, and many sperms were found in the tubules in phenylethanoid glycosides groups.Conclusion Phenylethanoid glycosides has obviously therapeutical effect on CTX-induced dyszoospermia in mice,and its mechanisms might be correlated with recovering the testosterone level.

Objective To observe the cellular morphologic features of coronary sinus orifice(CSO) and its peripheral myocardium and discuss its functional significance in radiofrequency ablation atrial reentrant arrhythmias and AVNRT. Methods A total of 7 out of 15 cases were observed by means of autopsy, and serial sections of 7 cases of adult hearts in sagittal plane were stained with HE and Mallory staining. The cellular features of different muscular fibers and their arrangement were observed under light microscope; one case was studied under electron microscope. Results CSO and the majority of coronary sinus were encompassed by atrial myocardium; CSO fibers toward posterior was connected with extension of terminal crest (CT); and supravalvular muscular ring of right atrium(RAMR) and that toward anterior was connected with AVN. P cells were seen in superior-anterior and inferior-anterior wall of CSO. A lot of T cells were discovered on the inferior wall of CSO. Purkinje cells were mainly found in the superior wall of CSO. The muscular fibers from CSO to AVN were composed of T cells and dissected into two parts: the first one was called right atrial nodular bundle and the second one posterior node extension which was identical with the ablation target of slow pathway. In addition, lots of nerve fibers were seen in myocardium of CSO adjacency particularly in the tissue of the posterior wall of CSO. Conclusion Myocardium of CSO adjacency may belong to slow pathway of DAVNP. The myocardium of CSO adjacency participates in the formation of circular pathway in atrial reentrant arrhythmias and AVNRT. It is possible that CSO is an important latent pacing maker.

Objective To observe the expression of vascular endothelial growth factor(VEGF) mRNA and protein in fluoride(F~-) treated fibroblast(FB) of mice in planar(2D) and FBs populated collagen lattice(3D) culture systems and to further explore the effects of VEGF on the osteogenic action of FB. Methods FB were divided into 0 (control group), 0.0001,0.0010,0.1000,1.0000,10.0000 and 20.0000 mg/L groups(F~-). The levels of VEGF mRNA and protein at 48 h were measured by using RT-PCR, ELISA and immunohistochemistry (IHC) methods. Results The expression of VEGF mRNA increased obviously in group of 0.1000 mg/L(1.08 ± 0.09) in 3D FB compared with the control group(0.93 ± 0.02, all P < 0.05). Fluoride increased the content of VEGF protein obviously in groups of 0.1000,1.0000,10.0000 mg/L(0.19 ± 0.02, 0.26 ± 0.01 and 0.32 ± 0.01 ), higher than that in 2D FB culture supematant in the control group(0.14 ± 0.01, all P < 0.05) ; and in groups of 0.1000, 1.0000 rag/L(0.59 ± 0.06 and 0.52 ± 0.03) it was higher than that in 3D FB culture supematant in the control group(0.37 ± 0.05, all P< 0.01 ). The IHC results showed that the VEGF positive staining cells increased significantly in group of 0.001 mg/L (0.45 ± 0.05) in 2D FB when it was compared with control group(0.36 ± 0.03, P< 0.05); and in groups of 0.0010, 0.1000, 1.0000 rag/L(0.62 ± 0.04,0.70 ± 0.06 and 0.65 ± 0.07) are it was higher than that in 3D FB control group (0.44 ± 0.04, P < 0.05 or < 0.01 ). Conclusions The higher expression of VEGF mRNA and protein in 2D and 3D FB induced by fluoride may play an important role in stimulating the osteogenesis ability in FB.

[Objective]To study the dental status,the development of jaw and the size of sella turcica of children with ectoder-mal dysplasia(ED).[Methods]Panaramic radiography and lateral cephalograms of six ED individuals(age range of 6~7 years,five males and one female)were obtained. The dental status was record. 17 measurements about the jaws and the sella turcica were mea-sured and compare them to Chinese children without ED syndrome.[Results]The mean number of missing teeth was 22.3 in perma-nent dentition and 16.2 in primary dentition;The teeth that most likely to absent were permanent lateral incisor ,maxillary first premo-lar,maxillary primary lateral incisor and mandibular primary central incisor,and all remaining teeth are in conical shape. Lateral cephalometric measurements showed that all ED subjects had lower ANS-Ptm,which suggested a short maxilla. Low Co-Po,ANB, NA-PA,N-Me,N-ANS and ANS-Me values that were found in all subjects,as well as low SNA,Y-axis,MP-FH,S-Co,and high SNB,NP-FH,NP-FH that were noted in some subjects showed counterclockwise rotation and protrusion of mandible with short-er length in ED subjects. Some subjects had low ANS-Me/N-Me × 100%and high N-ANS/N-Me × 100%,representing a short facial height. Five cases represented lower length and diameter of sella turcica;two cases showed lower depth of sella turcica ,indicating the abnormal development of sella turcica.[Conclusion]The results of this study suggest that the dentition ,jaws and sella turcica in ED children differs when compared to individuals without this syndrome.

Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P

AIM:To explore the effect of component II of broccoli polypeptide on the apoptosis in glioma cells . METHODS:Human glioma SHG-44 cells were cultured and divided into control group and 3, 10, 30 and 100 mg/L com-ponent II of broccoli polypeptide groups .Cell viability was detected by MTT assay .The apoptotic rates were examined by Annexin V/PI staining.The morphological changes of the cells were observed under inverted microscope .The protein ex-pression of Bax and Bcl-2 was detected by immunocytochemistry and Western blotting .The protein level of caspase-3 was also examined by Western blotting .RESULTS:Treatment with component II of broccoli polypeptide for 24 h, 48 h or 72 h induced significant inhibition of viability of SHG-44 cells in a time-and dose-dependent manner .The results of Annexin V/PI staining showed that the apoptotic rates were increased in treatment groups in a dose -dependent manner .The density of glioma cells was decreased after treated with increasing concentrations of the drug , and the apoptotic bodies were ob-served under inverted microscope at 72 h.The results of immunocytochemistry and Western blotting showed that the expres-sion of Bax protein was increased but Bcl-2 protein expression was decreased , and the ratio of Bax/Bcl-2 was increased sig-nificantly compared with control group (P<0.05 or P<0.01).The level of caspase-3 protein was increased in 30 and 100 mg/L component II of broccoli polypeptide groups compared with control group (P<0.01).CONCLUSION:The compo-nent II of broccoli polypeptide increases the ratio of Bax /Bcl-2 and activates caspase-3 protein, thus inducing the apoptosis of glioma cells.

AlM:To evaluate the clinical efficacy of intravitreal injection of ranibizumab combined with macular grid photocoagulation for diabetic macular edema ( DME) .METHODS:Totally 60 eyes ( 60 patients ) with DME were randomly divided into 2 groups: 30 eyes of simple injection group underwent intravitreal injection of ranibizumab, and 30 eyes of combined treatment group underwent intravitreal injection of ranibizumab and macular grid photocoagulation 1wk later. The best corrected visual acuity ( BCVA ) , central macular thickness ( CMT ) measured by optical coherence tomography ( OCT ) and postoperative complications were observed.
RESULTS:ln simple injection group, the BCVA after operation were separately 0. 390 ± 0. 075 (4wk), 0. 367 ± 0. 088 (8wk) and 0. 319 ± 0. 064 (12wk), the CMT after operation were separately 221. 63 ± 112. 34μm (4wk), 337. 73±99. 56μm (8wk) and 432. 92 ± 100. 46μm (12wk), which were much better than pre-operation. But during follow-up, the BCVA presented down trend and the CMT was on the rise slowly. ln combined treatment group, the BCVA after operation were separately 0. 385 ± 0. 036 (4wk), 0.382±0.079 (8wk) and 0.377±0.097 (12wk),the CMT after operation were separately 249. 77 ± 106. 55μm (4wk), 270. 40 ± 92. 88μm (8wk) and 275. 84 ± 97. 34μm (12wk ), which were satisfactory and steady during follow-up, better than simple injection group (P<0. 05).CONCLUSlON:lntravitreal injection of ranibizumab can effectively improve visual acuity and decrease central foveal thickness for patients with DME, combining with macular grid photocoagulation can ensure therapeutic effects steady and permanent.

Objective To detect the change of cerebral perfusion in pediatric patients with Leigh's syndrome (LS)by using MR perfusion technique.Methods Twelve patients with Leigh's syndrome and thirteen normal children were scanned with the sequence of flow-sensitive alternating inversion recovery exempting separate T_1 measurement (FAIREST).Their relative cerebral blood flow(rCBF)values were obtained in regions of bilateral basilar nuclei and bilateral thalami.Student t-test was used to compare them between the two groups and receiver operating characteristic(ROC)curve analysis was carried out.Results Statistical analysis revealed significant difference between two groups in the regions of bilateral basilar nuclei and right thalamus(t =3.26,P =0.002;t =2.25 ,P =0.018 ;t =2.88 ,P =0.004,respectively).The rCBF values for LS group and control group were 0.432?0.158 and 0.619?0.125 for right basilar nuclear, 0.478?0.186 and 0.621?0.123 for left basilar nuclear,0.630?0.189 and 0.833?0.160 for right thalamus,respectively.The areas under the ROC curves were 0.833 and 0.756 for the rCBF of right and left basilar nuclear,respectively.Conclusion Relative CBF maps may reveal changes of cerebral blood flow in some specific brain regions in patients with Leigh's syndrome.It can provide additional information to the clinicians in the evaluation of the disease.