Objective:To analyze the prognosis about periodontal ligament healing of replanted perma-nent teeth in children and to examine the associated factors.Methods: The sample consisted of 49 children with 61 avulsed permanent teeth, whose injuries had been managed in the period from 2000 to 2012.The clinical data of replanted teeth were collected, and the follow-up period was no less than 12 months .The factors were analyzed in relation to postoperative outcomes, classified as functional periodon-tal healing ( FH ) , infection-related ( inflammatory ) resorption ( IRR ) and replacement resorption ( RR) .Results:The functional healing rate was 23.0%, while replacement resorption rate was 72.1%. The replacement resorption ( ankylosis ) was usually observed earlier by clinical examination than by radiographic examination.86.0% (40/47) resorptive processes were diagnosed within the first year. Physiological storages, such as milk, saline and saliva were significantly better to periodontal ligament healing than nonphysiological storages, such as tap water and sterilizing solutions ( chloramine and alco-hol) .Functional healing was found significantly more frequent in canines and premolars.Conclusion:The factor significantly affecting periodontal ligament healing is storage medium.Replacement resorption is the most common type of root resorption.The replacement resorption diagnosis must combine the radio-graphic examination with the clinical examination.It is better to follow up more than 1 year after tooth re-plantation.

OBJECTIVETo investigate the functions of human periodontal myofibroblast (MFB) in vitro.

METHODSHuman periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.

RESULTSThe experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).

CONCLUSIONHuman periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.

Actins ; Epithelial Cells ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Jaw ; metabolism ; Myofibroblasts ; Transforming Growth Factor beta1

Objective:To analyze the pulpal prognosis of replanted permanent teeth in children and to examine the associated factors .Methods:The samples consisted of 67 children with 81 avulsed perma-nent teeth treated in Peking University Hospital of Stomatology from 2000 to 2012 .Their clinical data of the replanted teeth were collected , and the follow-up period was no less than 12 months.The pulpal prognosis was classified as pulp necrosis and pulp non-necrosis .Results: The replantation within 30 minutes after avulsion led to the most significant increase in pulpal healing (P<0.05).The dental pulp of 80% ( 4/5 ) young permanent teeth replanted within 30 minutes remained vital , while all the teeth replanted after 30 minutes developed pulp necrosis within 60 days after replantation .Conclusion: The extra-alveolar period significantly affects the prognosis of pulp in immature permanent teeth after replanta-tion.When the extra-alveolar period is more than 30 minutes, the chance of pulp revascularization after replantation is very low , therefore pulp extirpation should be performed .

Objective To study the expression of human giant larvae-1 (Hugl-1) in ovarian carcinoma and its clinical significance.Methods Hugl-1 mRNA expression in 31 ovarian cancer and the corresponding adjacent tissues was examined by real time fluorescence quantitative RT-PCR and in situ hybridization.Moreover,analysis was done while taking into consideration of the clinicopathologic parameters of ovarian cancer.Results The expression of Hugl-1 mRNA in ovarian cancer tissue was significantly higher than that in the corresponding adjacent tissues ([0.051 ? 0.029]vs [0.026 ? 0.043],P

This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

Objective To observe the effect of different needling retaining times in clinical curative effect of acupuncture on the treatment of periarthritis humeroscapularis.Methods A total of 60 patients, who met the inclusion criteara, were randomly divided into 2 groups according to random number table method, 30 patients in each group. Both groups were treated with the same acupuncture therapy for 20 times, but the needling retaining times were different as 20 minutes and 40 minutes. The curative effect rates, Visual Analog Scale (VAS) and Constant-Murley Score (CMS) were evaluated.Results After the treatment, the VAS scoresin 40mins group was significantly lower than that in the 20 mins group (2.67 ± 1.03vs.3.60 ± 1.48,t=-3.251, P<0.01); the CMSin 40 mins group was significantly higher than the 20 mins group (73.20 ± 10.88vs.66.47 ± 12.62,t=-2.199,P<0.05). The curative effect rates of the 40 mins group was 26.7% (8/30), which was significantly higher than 3.3% (1/30) in the 20 mins group (χ2=4.706,P=0.030). The total effective rates in the 40 mins group was 90.0% (27/30), which was significantly higher than 96.7% (29/30) in the 20 mins group (χ2=0.268,P=0.605).Conclusions Acupuncture treatment for the patients with periarthritis humeroscapularis showed that the 40 minutes of needling retaining times had better symptom improvement and restore function effects than 20 minutes, however the total effective rate was no significant difference.

Objective To construct eukaryotic expression plasmid of hCD137 Fc gene and express hCD137 Fc fusion protein with high biological activity. Methods PCR technique was employed to clone the human CD137 cDNA from a normal human activated T cells cDNA library, and then clone its extramembrane encoding region. The extramembrane sequence together with human IgG1 Fc cDNA were inserted into the eukaryotic expression plasmid pcDNA3. CD137 Fc gene was expressed transiently in 293T cells, and then CD137 Fc protein was purified by recombinated protein A affinity chromatography column. At last, the MW, purity and antigenicity of CD137 Fc were identified by sandwich ELISA, SDS PAGE and Western blotting, respectively. Results The ORF of CD137 Fc gene was coincident with what we expected. ELISA and SDS PAGE confirmed protein expression in 293T cells. Western blotting proved the antigenicity of the purified CD137 Fc protein. Conclusion We obtained a purified recombinated CD137 Fc protein with biological activity and the expected MW. This lays the foundation for further studies of CD137 such as its role in immune homeostasis and other biological functions.

Objective To clone the full length cDNA of human LIGHT and to construct the recombinant eukaryotic expression plasmid pCI neo LIGHT for the stable expression on 293T cells. Methods Human LIGHT cDNA was cloned from a normal human activated T cell cDNA library phAD.CAD by PCR. After sequencing, LIGHT cDNA was inserted into plasmid pCI neo for the construction of the eukaryotic expression. The recombinant was transfected into 293T cells by electroporation. The expression of LIGHT on the surface of 293T cells was detected by flow cytometry after screening with G418. Results Sequencing confirmed that ORF of LIGHT gene was intact and right. Restrictive enzyme digestion proved that LIGHT gene was inserted into the recombinant plasmid of LIGHT pCI neo correctly. FACS analysis revealed that about 78.69% 293T cells expressed LIGHT protein on the cell surfaces at 3 months after screening with G418. Conclusion LIGHT gene has been cloned successfully, and a 293T cell line expressing LIGHT protein on its membrane surface has been obtained.

CD59 antigen is a widely expressed cell surface glycosylphosphatidyl-inositol (GPI) anchored glycoprotein.It acts as an inhibitor to the assembly of the membrane attack complex of homologous complement,binds to CD2,and also transduces activation signals with T cells.In this report,a 396bp DNA fragment was amplified by RT-PCR method from the total RNA of Jurkat cells.The fragment was cloned into pUC18 and pUC19 plas-mids,and further sequenced by Sanger′s-dideory-mediated chain termination.The results showed that this cDNA fragment included 384bp open reading fragment and its sequence was identical to the published sequence encoding human CD59 antigen.Furthermore,the cDNA of CD59 was subcloned into retroviral vector pLXSN and transfec-ted into packaging cell line PA317 to generate stable virus-producing cell lines.Then,mouse thymotase cell line EL-4 and fibroblasts cell line NIH3T3 were infected with the virus resulting in stable expression of CD59 on the cell surface.The transfected cells were tested for their susceptibility to human complement-mediated cytolysis.It was found that the transfected cells expressing CD59 antigen were far less susceptible than the controls,indicating that the gene for CD59 can be expressed in xenotypic cells stably to confer protection against human serum complement.

Objective: To investigate the MRI manifestations of primary testicular lymphoma (PTL). Methods: Clinical and MRI data of 8 patients with pathologically confirmed PTL were retrospectively analyzed. Results: Plain MRI showed morphological abnormalities of affected testis in all 8 cases, and the volume increased. The lesions appeared as masses with clear borders, present low signals on T1WI in all 8 cases and spot-like high signals in 3 cases, uneven iso-low signals on T2WI showed. Epididymis and spermatic cord involvement were found in 3 cases, the boundary of testis and epididymis were not clear, and signals were heterogeneous, spermatic cord with multiple curved strips of high signal on T2WI. Hydrocele occurred in 3 cases, including 1 case involved epididymis and spermatic cord. Obviously heterogeneous enhancement of lesions was found during delayed enhancement scanning, and unenhanced areas were noticed in 3 cases. The time-intensity curves (TIC) of lesions presented outflow type (type III) in all 8 cases. On DWI, the solid parts of lesions showed high signal, which showed low signal on ADC map in all 8 cases. Conclusion: MRI manifestations of PTL had certain characteristics. Combination of plain and dynamic enhancement MR were helpful to correct diagnosis of PTL.