BACKGROUND: In the tooth regeneration and dental tissue engineering study, finding suitable stem cells are the central issues.OBJECTIVE: To isolate and culture child exfoliated deciduous tooth pulp stromal cells, and compare difference in dentin sialophosphoproteln expression prior to and following mineralization.DESIGN, TIME AND SETTING: The cell observational experiment was performed at the Central Laboratory of Daqing Oil Field General Hospital from December 2006 to December 2007.PARTICIPANTS: Dental pulp were obtained from exfoliated deciduous molar tooth of eight children aged from eight to ten, four males, four females. The possibility of infectious diseases, endocrine disease, dental caries and periodontal disease was excluded.METHODS: Deciduous teeth pulp stromal cells were obtained by tissue block adhesion method, and then cultured in DMEM with 10% fetal bovine serum for 14 days. Following digestion and passage, cells were incubated in DMEM/F12 supplemented with 50 pmol/L laevo-ascorbic acid, 10 nmol/L vitamin D3. 10 mmol/L Na β-glycerophosphate and 10-8 mol/L dexamethasone for another 14 days. Cells cultured in complete DMEM served as controls.MAIN OUTCOME MEASURES: Cell morphology and growth rule were observed under a microscope. Difference in dentin sialophesphoproteln expression was determined prior to and following mineralization using immunocytochemical staining.RESULTS: Dental pulp stromal cells from human exfoliated deciduous tooth, adhered to the wall, were polygon-shaped and characterized by large cell size and a relatively large nucleus and plenty cytoplasm after mineralization for 14 days. Over 80% cells were positive for dentin sialophosphoprotein. Non-induced cells were spindle, and only 2% cells were positive for dentin sialophosphoprotein.CONCLUSION: Odontogenic inductive culture might improve odontoblastic differentiation. It seems that human exfoliatod deciduous tooth pulp represents an new reservoir of adult stem cells with odontogenic potential.

Objective To study the feasibility and clinical experience of off-pump multivessel coronary artery bypass grafting (OPCAB) for patients with acute myocardial infarction (AMI). Methods Sixty-one patients with AMI were treated surgically. The mean age of was (64.7?7.2) years. The last attack angina pectoris unreliable with use of Nitro-Glycerine, and the levels of CK-MB documented (3.5?1.8) times and TnI were (10.9?4.1) times higher than the normal. OPCAB was performed in sixty-one patients and in two patients, converted to cardiopulmonary bypass. All patients were followed up from twelve months to twenty-four months. Results The mean interval between the onset of AMI and the accomplishment of OPCAB was (115.8?15.1) hours ,and the number of distal anastomosis was (3.4?0.7)/pt. The mortality rate was 3.28%. During the period of follow-up, all patients were asymptomatic and the results of echocardiography showed the function of ventricles improved. Conclusion Off-pump technique applying to multivessel coronary artery disease with AMI yielded satisfied clinical outcome. The morbidity and mortality of OPCAB is substantially lower maybe due to avoiding the adverse effects of CPB.

Objective to measure the percentage of th22 cells and evaluate the levels of plasma interlukin-22(IL-22)in human peripheral blood, so as to determine their clinical significance in primary Sj?gren′s syndrome(pSS). Methods Patients with pSS were divided into three subgroups based on severity of labial gland involvement:mild,moderate and severe. Healthy people served as controls. the percentage of th22 cells and the lev-els of IL-22 in human peripheral blood from pSS patients and healthy controls were measured and compared. Results Compared to healthy con-trols,pSS patients had significantly higher percentage of th22 cells and higher plasma IL-22 levels(P < 0.05). Among pSS patients,the more seri-ous illness they had,the higher percentage of th22 cell and levels of IL-22 were observed. Severe patients had higher percentage of th22 cells and IL-22 levels than moderate patients(P < 0.05),and moderate patients had higher percentage of th22 cells and IL-22 levels than mild patients(P <0.05). Conclusion Increased peripheral IL-22-secreting th22 cells are detected in primary Sj?gren′s syndrome,which have a close association with disease severity. these data suggest that th22 and its released cytokine IL-22 may be considered as potential valuable biomarkers for severity of pSS,which may provide a novel therapeutic target for treatment.

Objective To explore the efficacy of multiple target therapy in treatment of patients with chronic moderate and severe IgA nephropathy with hyperuricemia.Methods Seventy-six patients with chronic progressive moderate and severe IgA nephropathy with hyperuricemia were enrolled the current study and randomly divided into observation group and control group.Patients in control group were treated with allopurinol,prednisone,benner pury and valsartan,while those in observation group were treated with urokinase,mycophenolate mofetil besides the basis of control group for 6 months.The blood uric acid (UA),24 h urine protein,mean arterial pressure (MAP) and creatinine clearance rate (Ccr) were determined and analyzed.Results The levels of UA,24 h urinary protein,MAP and Ccr in observation group and control group were same before treatment (P ＞ 0.05).After 6 months treatment,the levels of UA,24 h urine protein,MAP and Ccr in observation group were (413.7 ± 90.7) μmol/L,(1.15 ± 0.57) g/L,(87.7 ± 10.6) mmHg and (81.9 ± 3.7) ml/min respectively,significantly different from those of the control group ((369.6 ± 67.2) μ mol/L,(0.77 ±0.51) g/L,(81.6 ±12.3) mmHg and (86.4 ±6.8) ml/min;t =2.219,2.802,2.132,3.230;P ＜0.05).The rate of adverse reactions in two groups was not significantly differnent(9.7％ (3/31) vs 9.1％ (3/33) ; x2 =0.006,P =0.936).Conclusion Multiply target therapy is effective and safe in terms of treating chronic progressive moderate and severe IgA nephropathy with hyperuricemia.

Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

The effects of Panax notoginsen saponins ( PNGS ) on myocar- dial ischemia and reperfusion injury in conscious rabbit were studied with observation of changes in electrocardiogram (ECG), the activities of creatine phosphokinase ( CPK ) and lactate dehydrogen-ase ( LD ) and the size of ischemic area. 50 mg/kg and 100 mg/kg PNGS significantly decreased the activities of CPK and LD, and the abnormal changes of ECG during ischemic and reperfusion periods. Also, PNGS significantly reduced the size of myocardial ischemic area. These results suggest that PNGS have the protective effects on myocardial ischemia and reperfusion injury.

AIM: To prepare a soluble hemangiopoietin(HAPO) protein and to construct pET22b(+) expression vector, to obtain pure recombinant HAPO protein and to measure its bioactivity. METHODS: HAPO cDNA was amplified using RT-PCR method from a commercial human fetal liver cDNA library. The resulting product was cloned into pET22b(+) vector and transformed into E.coli BL21(DE3). The recombinant protein was isolated and purified by Ni~(2+)-NTA chelating resin and the chromatographies of SP Sepharose FF. The adhesion of human umbilical vein endothelial cells (HUVEC) were measured by adhesion assay. RESULTS: HAPO gene with a reading frame of 897 bp was successfully cloned from human fetal liver cDNA library, the expressed pET22b(+)-HAPO fused protein existed in a soluble form, with the yield above 10% total bacterial protein and its purity achieved above 80%. The activity assay showed that the treatment of HAPO enhanced total adherence of HUVEC in a concentration-dependent manner. CONCLUSION: HAPO protein can be expressed in a soluble form. HAPO may facilitate the homing of hematopoietic stem/progenitor cells in vitro.

Objective To investigate the diagnosis and therapy of primary maligmant tumors of the heart. Methods Clinical data of 21 patients with malignant cardiac tumors admitted to our department from June 1980 to May 2008 was analyzed and the references were reviewed. Results All patients received operations. Pathological classification of the tumors was made by histological examinations. Radical resections for 10 eases and partial resections for 5 eases were performed. The other 6 patients received only thoracotomy and cardiac exploration. Three eases were lost during follow up. Three survivors received radical resections are still alive now 2-15 months after the surgery, while all the other patients died within 4 years after the operation due to malignant tumor recurrence and (or) metastasis. Conclusion Echocardiography, CT, 3D-CT, MRI, coronary CT and angiocardiography are helpful for the diagnosis of the malignant cardiac tumors and the selection of operations. Histological examination is necessary for the final diagnosis. Early diagnosis, radical resection and post-operative radiotherapy and chemo therapy may provide a better result.

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .