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.

Objective To evaluate the safety and efficacy of thoracoscopic lobectomy for pulmonary benign diseases.Methods Between July 2002 and September 2007,35 cases with pulmonary benign diseases underwent thoracoscopic lobectomy in our hospital.Of the patients,video-assisted thoracoscopic lobectomy was performed on 12 cases,and totally endoscopic lobectomy was carried out in 23.Results The operation was completed in all of the cases except in one who was converted to open surgery because of massive hemorrhage.In this series,no peri-operational death occurred,however,complications occurred in 3 cases(morbidity rate: 8.6%),including 2 cases of persistent air leak and 1 case of pneumonia.The mean duration of chest tube drainage was 3.6 days(2 to 7 days),and average hospital stay after operation was 7.7 days(2 to 14 days).Postoperative pathological diagnosis included bronchiectasis in 15 patients,pulmonary inflammatory pseudo-tumor in 6,tuberculosis in 5,fungal infection in 5,pulmonary sequestration in 2,and bronchogenic cyst in 2.Conclusions Thoracoscopic lobectomy is safe and effective for pulmonary benign disease.

Endoplasmic reticulum(ER) is a locus where proteins are modified,folded and calcium stored.The accumulation of unfolded protein and disorder of calcium ion can lead to ER stress.Early ER stress will induce unfolded protein response(UPR)which may be protective to cells.On the contrary,long-time strong ER stress will induce cell apoptosis,even death.ER stress has been suggested to be involved in some human neurodegenerative diseases.However,the exact contributions of ER stress in the various disease processes are yet unknown.This review mainly summarizes recently reported discoveries concerning ER stress associated with neurodegenerative diseases and highlights current knowledge in this field that may reveal novel insight into disease mechanisms and help to design better therapies for these disorders.

Objective To investigate the possible mechanisms of enhancing angiogenesis and arteriogenesis through the oberservation of the effect of gene transfer of angiopoietin-1 (Ang1) on its receptor Tie2 in rat models of acute myocardial infarction. Methods Myocardial infarction was induced in rats by left anterior descending artery ligation. Naked plasmid DNA encoding human angiopoietin-1 (phAng1) was delivered into the ischemic area (group A) by intramyocardial injection. On day 3, 7, 14 and 28 after the injection, the mRNA expression of Tie2 and its changes with time were determined by RT-PCR. The number of vessels and arterioles was examined by immunohistochemistry. The collagen was evaluated by Masson staining. Results RT-PCR showed that mRNA expression levels of Tie2 in group A were significantly higher than those in the control group, reached the highest level on day 7 post-injection, and gradually declined to normal level 28 days later. On day 7, 14 and 28, the vessel count showed the number of blood vessels (angiogenesis and arteriogenesis) in group A was greater than that of the control group at the same timepoint and the infracted myocardium in group A was significantly less than that of the control group. Conclusion Gene transfer of phAng1 enhances angiogenesis and arteriogenesis in acute myocardial infarction and reduces the infraction area probably by upregulating the expressions of Tie2 receptor.

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 value of the use of the vacuum sealing drainage technology (VSD) in the dermatoplasty for large area of cutaneous defects.Methods By a prospective randomized study,64 patients with large area of cutaneous defects were randomly divided into traditional packaging group (dermatoplasty with the traditional dermatoplastic package method,n =32) and VSD group( dermatoplasty with the vacuum sealing drainage technology,n =32).Operating time,survival.rate of skin graft and wound infection were compared between these two groups.Results There was no significant difference in the operation time between the two groups( [77 ± 16]min vs [83 ± 12] min,P =0.06).The survival rate of skin graft and wound infection in VSD group were significantly better than those in traditional packaging group( survival rate of skin graft:(6±1)％ vs (20±7)％,P＜0.001;wound infection cases:1 vs6,P=0.039).Conclusion Treating large area of cutaneous defects by the dermatoplasty with VSD technology could significantly improve clinical efficacy.

Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.

Objective To investigate experience in nursing the patients with emergency traumatic injury combined with postoperative atelectasis. Method The clinical data of 11 patients with emergency traumatic injury combined with postoperative atelectasis were reviewed for summarizing the nursing experience. Result The clinical symptoms of all the 11 patients disappeared and the lungs reexpanded. Conclusion Careful observation of the disease conditions in order to prevent and treat atelectasis by airway humidification, sputum drainage and early exercises are effective for the cure of postoperative atelectasis.

Purpose To investigate the expression of RNF2 in breast disease tissues and cell lines,and to analyze the association between expression of RNF2 and clinicopathological characteristics in breast carcinoma.Methods Expression of RNF2 protein and mRNA levels was detected using immunohistochemistry of EnVision two-step and qRT-PCR in breast carcinoma and benign breast disease as well as in cell lines.Results RNF2 expression was sigmificantly higher in breast carcinoma tissue specimens compared with benign breast disease specimens (P ＜0.05).Besides,the expression of RNF2 protein was significantly associated to tumor size,lymph node status and TNM stage (P ＜ 0.05 for both),but was not related to age,histological grade,the expression of ER,PR and HER-2 (P ＞ 0.05 for both).Higher expression of RNF2 mRNA was detected in breast carcinoma cell lines compared with breast epithelial cell lines (P ＜ 0.05).Conclusion RNF2 is overexpressed in breast carcinoma and can be a potential therapeutic target for breast carcinoma.

AIM To study the protective effects of prostaglandin E 1 on hypoxia/reoxygenation injury in cultured neonatal rat myocytes. METHODS The hypoxia/reoxygenation injury model of cultured neonatal rat myocardial cells was developed. The activities of plasma superoxide dismutase (SOD) were measured by the method of xanthine oxidase and the contents of serum malonicaldehyde (MDA) were determined by the method of thiobarbituric acid. The activities of dehydrogenase in mitochondria were detected by MTT assay. The activities of LDH in culture were also evaluated. CONCLUSION PGE 1 has protective effects on the cultured neonatal rat myocardial cells injured by hypoxia/reoxygenation. The mechanisms are related to its antioxidation effects.