BACKGROUND: Presently used biological factors for inducing bone marrow mesenchymal stem cells (BMSCs) do not obtain mature chondrocytes. Cartilage tissue engineering using BMSCs as seeds does not collect tissue-engineered cartilage, which has value in the clinic. The obtained tissue is cartilage-like tissue. OBJECTIVE: To screen differentially expressed genes between rat BMSCs and chondrocytes by microarray. DESIGN, TIME AND SETTING: The cytology gene study was conducted at the Central Laboratory, First Affiliated Hospital, Dalian Medical University in July 2007. MATERIALS: A total of 8 healthy Sprague Dawley rats aged 2 months were obtained from Animal Experimental Center, Dalian Medical University. 27K Rat Genome Array chip was supplied by Bo’ao, Beijing, China. METHODS: Rat BMSCs and chondrocytes in the aural region were sterilely isolated and cultured in vitro. Total RNA was extracted and purified using Trizol one-step method, and transformed into double chain cDNA probe following reverse transcription. Cy5-dCTP and Cy3-dCTP were used to label BMSCs and chondrocytes, which were hybridized and washed. The fluorescent signals were scanned by a scanner. The values were analyzed and calculated by GenePix Pro 4.0 software. MAIN OUTCOME MEASURES: Gene expression spectrum chip hybridization results. RESULTS: Among the differentially expressed genes (2 times difference), BMSCs as controls, upregulated and downregulated genes were 1 226 and 888, respectively. There were many differential expression gene of BMSCs and chondrocytes. Cy5/Cy3 20 genes are defined as significant differential expression gene. Thus, two important cytokines were found: chondromodulin and connective tissue growth factor. CONCLUSION: The gene chip technique provides an ideal method for screening cytokines during study of tissue-engineered cartilage. Cartilage regulin and connective tissue growth factor highly express in chondrocytes, which indicated that the two have closely association with the differentiation of BMSCs into cartilage.

10.3969/j.issn.2095-4344.2013.23.007

BACKGROUND:Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰmaybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately.
OBJECTIVE:To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells.
METHODS:Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in celllines.
RESULTS AND CONCLUSION:The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successful y constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells.

Objective To explore the adverse effects of formaldehyde(FA)on learning and memory ability of mice and the antagonistic effect of N-acetyl-cysteine(NAC),an antioxidant.Methods Thirty-four ICR mice were randomly divided into four groups,the control(NS,n=8),treated with FA(15 mg/kg,n=9),treated with NAC(100 mg/kg,n=8),treated with FA(15 mg/kg) plus NAC(100 mg/kg,n=9),the treatment was conducted by intraperitoneal injection once a day for seven consecutive days.On the eighth day,the learning and memory ability were tested by using water labyrinth task for seven consecutive days.Results The mice in FA group behaved excited,restless and then turned to repose,moveless and clustering,but this phenomena was not seen in the other groups.There was no significant difference in the body weight of mice among groups.As for learning,latent period in the FA group [(27.15?2.66)s] was significantly longer than that in the control group [(15.83?2.82)s] and the FA+ NAC group[(14.98?2.66)s],and revealed statistical significance(P

The tools used for the literature quality evaluation are introduced. The common evaluation tools that are publicly and extensively used for the evaluation of clinical trial literature quality in the world are analyzed, including Jadad scale, Consolidated Standards of Reporting Trials (CONSORT) statement and Grades of Recommendations Assessment, Development and Evaluation (GRADE) system and the others. Additionally, the present development, updates and applications of these tools are involved in analysis.
Biomedical Research ; standards ; Evaluation Studies as Topic ; Humans ; Publications ; standards ; Quality Control

Objective To compare the effect of transthoracic contrast echocardiography (cTTE) with transesophageal contrast echocardiography (cTEE) for detecting right to left shunt (RLS) in patients with patent foramen ovale (PFO). Methods The prospective study was conducted in 29 consecutive patients with PFO who suffered from cryptogenic stroke and/or migraine. Contrast echocardiography was performed in all 29 patients. The cTTE was performed using transducer with second harmonic imaging modality (transmitting frequency 1.7 MHz, receiving frequency 3.4 MHz). The cTEE was performed using transducer with frequency 7 MHz. Ten milliliter saline solution of contrast were rapidly administrated through an antecubital vein. According to whether microbubble (MB) appearing in left atrium after complete opaciifcation of right atrium within the ifrst 3 circles, the results were classiifed by a four-level semi-quantitative categorization:Level 1 (no PFO-RSL), no MB in left atrium; Level 2 (small PFO-RSL) 1-10 MBs; Level 3 (medium PFO-RSL) 10-30 MBs;Level 4 (large PFO-RSL)>30 MBs. Results The total detection rate of PFO-RSL was signiifcant different between cTTE and cTEE (86.2%vs 55.2%,χ2=5.711, P=0.017). In cTTE there were 4 cases at level 1, 1 case at level 2, 5 cases at level 3 and 19 cases at level 4. In cTEE there were 13 cases at leverl 1, 2 cases at level 2, 6 cases at level 3 and 7 cases at level 4. The comparison of semi-quantitative grading derived from cTTE and cTEE was also signiifcant different (Wilcoxon signed ranks test showed Z=-3.789, P=0.000). Conclusions The efifciency in detection of PFO-RLS with cTTE was better than with cTEE. Compared with cTEE, cTTE was easier in practice and brought less discomfort and complications to patients.

Objective To discuss the feasibility of constructing the system of traditional Chinese medicine nursing diagnosis.Methods Based on the theoretical analysis and status quo analysis,the feasibility of constructing the system of TCM nursing diagnosis was discussed,and the achievements and problems waiting for settlement were also narrated.Resuts It has the foundation of constructing the system of TCM nursing diagnosis,but some problems still need to be solved.Conclusions It is feasible for building TCM nursing diagnosis system,and the TCM nursing diagnosis system does not conflict with NANDA-I.

Female ; Humans ; Liver Neoplasms ; Middle Aged ; Paraganglioma

Objective To establish a closed-chest pulmonary embolism-reperfusion animal model by Swan-Ganz catheter and to explore the mechanisms of pulmonary embolism(PE)-reperfusion injury(RI). Methods Experiments were made on 14 mongrel dogs,ranging in weight from 15 to 18 kg,anesthetized with 3% pentobarbital sodium.The dogs were intubated with I.D.7 endotracheal tubes.Under sterile conditions,a 7 F Swan-Ganz catheter via the external jugular vein was positioned in the unilateral pulmonary diaphragmatic lobe(DL)artery.Occlusion/reperfusion of the DL artery was controlled with 1.2 ml diluted contrast agent filled into/drawn from the balloon.After the 24 h PE,the balloon was deflated to result in 4 h reperfusion of the DL.Measurements of blood gases and tumor necrosis factor-?(TNF-?)were made at normal condition,at 24 h PE and at 4 h reperfusion.Thin-section CT scans were performed at normal condition,24 h PE,30 min,1,2,3 and 4 h reperfusion,respectively.At the end of each experiment, tissue specimens of bilateral diaphragmatic lobes were obtained for both wet/dry(W/D)weight ratio and for pathological study.Results Reperfusion pulmonary edema(RPE)was an acute,mixed,noncardiogenic edema that was observed in all 14 dogs who had been successfully established as PE/RI animal models.RPE demonstrated heterogeneous ground-glass opacifications that predominated in the areas distal to the recanalized vessels.It manifested pathologically as an edematous lung infiltrated by inflammatory cells.The mean ofPaO_2 and TNF-? of 4 h reperfusion was(81?4)mm Hg(l mm Hg=0.133 kPa)and(16.0? 2.5)pg/ml,which were significantly different(P

Objective To provide a basis for specific intervention by dynamic observation of the households'related health life behavior and the children fluorosis change trend caused by the coal-burning-borne endemic fluorosis in Pu'an County.Methods Three villages were chosen using simple random sampling method in 2011-2013 for continuous monitoring.Every monitoring year in each monitoring village,according to the principle of every household survey,10 households were selected to survey related health life behavior;,and in the monitoring village all the children aged 8-12 years old were selected for Diagnosis of Dental Fluorosis (WS/T 208-2001),and in each age group,10 urine samples (half male half female) were collected,the fluoride content was determined by the ion selective electrode method (WS/T 89-1996).Results In 2011-2013,the households stoves qualified rate,stoves correct usage rate and the corn or chili correct drying rate were all 100.00％ (30/30).The dental fluorosis incidences of 8-12 years old children were 63.60％ (332/522),25.44％ (173/680) and 14.29％ (80/560),respectively,decreased year by year (x2 =324.67,P ＜ 0.05).In 2011-2013,children urinary fluoride levels were 0.83,0.84 and 0.68 mg/L,respectively.The difference between years was statistically significant (x2 =26.89,P ＜ 0.05),and the level in 2013 was significantly lower than those of 2011 and 2012 (all P ＜ 0.01).Conclusion In 2011-2013,the residents in Pu'an have good health life behavior,and the condition of children's dental fluorosis is decreased year by year.