Objective:To investigate the exression of miR-34a on lung cancer and normal lung tissues,and the effect and mechanism of miR-34a in lung cancer cell invasion and migration.Methods: qPCR was used to detect the expression of miR-34a on lung cancer.miR-34a-mimic and miR-34a-inhibitor were used to overexpress and knockdown miR-34a.qPCR was used to detect the effectiveness.Western blot was used to detect the expression of Snail after induced with miR-34a-mimic and miR-34a-inhibitor.Luciferase reporter gene was used to detect interaction between miR-34a and Snail.Transwell invasion assay was used to detect invasion ability after induced with miR-34a-mimic and miR-34a-inhibitor.Scratch assay was used to detect migration ability after induced with miR-34a-mimic and miR-34a-inhibitor.The expression of E-cadherin,Vimentin and Twist were detected by Western blot.Results: miR-34a expression was significantly reduced in lung cancer.With the stage of lung cancer progression,the expression of miR-34a reduced.With the differentiation of lung cancer progression,the expression of miR-34a decreased.Decreasing of miR-34a was associated with lung cancer lymph node metastasis.miR-34a-mimic and miR-34a-inhibitor could overexpress and knockdown miR-34a.miR-34a could regulate expression of Snail.Snail was the direct target of miR-34a;miR-34a could regulate the invasion ability of human lung carcinoma H1650 cells;miR-34a could regulate the migration of human lung carcinoma H1650 cells;miR-34a could regulate the expression of E-cadherin,Vimentin and Twist.Conclusion: miR-34a plays the role of tumor suppressor factor in lung cancer.miR-34a can regulate the invasion and migration ability of lung carcinoma H1650 cells by Snail induced EMT.

Objective:To investigate the inhibitory effect of perillyl alcohol (PA) on the proliferation and invasion of tung cancer cell A549,and the influence of PA on tumor angiogenesis was studied.Methods:Different concentrations of PA and erlotinib were added into lung cancer cell A549,the inhibiting effect of drug group on lung cancer cell A549 was found by MTT assay.The inhibiting effect of PA on lung cancer cell A549 invasion was measured by Transwell assay.ROS changes of PA on lung cancer cell A549 was detected by fluorescent.Influence of PA on Caspase-3 activity of lung cancer cell A549 was measured by spectrophotometry,VEGF,HIF-1 α,COX-2 expression in lung cancer cell A549 was measured by Western blot,and the NF-κB activity of lung cancer cell A549 was measured by EMSA.Results:Compared with blank control group,cell growth inhibition rate of PA and erlotinib on lung cancer cell A549 was increasing with the increased concentrations (10,50,100 μ,g/ml),the difference was statistically significant (P＜ 0.05),the invasion ability of lung cancer cell A549 was decreased continuously,the difference was statistically significant (P＜0.05).The ROS level of lung cancer cell A549 had no obvious change with the increasing density of erlotinib,but obviously increased with the increasing concentrations of PA (10,50,100 μg/ml).With the increasing concentrations of PA,the expression of COX-2,VEGF and HIF-1α were continuously decreased.EMSA assay showed that NF-κB was continuously decreased with the increasing concentrations of PA.Conclusion:The antitumor mechanism of PA on lung cancer cell A549 might be related to increase the expression level of ROS and reduce the expression of activity of NF-κB,COX-2,VEGF and HIF-1α with angiogenesis signaling pathway.

BACKGROUND:Bone marrow mesenchymal stem cel transplantation is considered as a promising therapy for spinal cord injury. How to more effectively promote the survival of bone marrow mesenchymal stem cel s in the area of spinal cord injury and to accelerate the recovery of motor function after spinal cord injury is a current study focus. Previous studies have found that low-frequency electromagnetic fields can promote bone marrow mesenchymal stem cel proliferation and differentiation, but whether the low-frequency electromagnetic fields can be applied to bone marrow mesenchymal stem cel transplantation for treatment of spinal cord injury requires further studies. OBJECTIVE:To discuss the effects of low-frequency electromagnetic fields on motor function of spinal cord injury rats after transplantation of bone mesenchymal stem cel s. METHODS:Sixty-four rat models of incomplete spinal cord injury at T 10 were established by compression method and then randomized into control group, transplantation group (bone mesenchymal stem cel transplantation), electromagnetic field group and combination group (electromagnetic field+bone mesenchymal stem cel transplantation). After successful modeling, bone mesenchymal stem cel s labeled with 5-bromo-2'-deoxyuridine were injected into the original injured site in the transplantation group and combination group, which were isolated and purified with the fast adherence method;while alpha-minimum essential medium was injected into the electromagnetic field group and control group for instead. At 24 hours post-operation, the electromagnetic field group and combination group were explored to low-frequency electromagnetic fields (frequency 50 Hz, magnetic indaction intensity 5 mT) for 60 minutes per day. RESULTS AND CONCLUSION:After cel transplantation for 21 days, the Basso, Beattie, and Bresnahan scores in the combination group was higher than the other groups (P<0.05). 5-Bromo-2'-deoxyuridine positive cel s grew wel , and integrated into the normal spine;syringomyelia was reduced, and the number of spinal neural cel s was increased in the combination group. In addition, glial fibril ary acidic protein expression was decreased in the combination group, while matrix metal oproteinase 2 expression was increased. It indicates that low-frequency electromagnetic fields could promote recovery of motor function in the spinal cord injury rats transplanted with bone mesenchymal stem cel s, which could be associated that low-frequency electromagnetic fields facilitate the survival of transplanted bone mesenchymal stem cel s, up-regulate the expression of matrix metal oproteinase 2, and reduce glial scar formation in the spinal cord injured site.

BACKGROUND:The repair and management of ful-thickness skin defects resulting from burns and chronic wounds remain a significant unmet clinical chal enge. Using epidermal stem cel s and keratinocyte growth factor for ful-thickness wound repair is a promising approach. Low-frequency electromagnetic fields which are a non-invasive physical stimulation therapy have been recognized as a good method to enhance wound healing. OBJECTIVE:To develop a new strategy to accelerate wound healing by transplanting transfected epidermal stem cel s and keratinocyte growth factor and treating with low-frequency electromagnetic fields in a mouse model. METHODS:Epidermal stem cel s from Sprague-Dawley neonatal rats were isolated and cultured in vitro, then the cel s were labeled with 5-bromo-2-deoxyuridine and transfected by Ad-KGF, a recombinant adenovirus carrying the keratinocyte growth factor. Mice were given to create ful thickness skin wound on the dorsum and randomly assigned to four groups:control group, transplantation of epidermal stem cel s group, transplantation of keratinocyte growth factor gene modified epidermal stem cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group. RESULTS AND CONCLUSION:The best healing pattern was observed in the keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group (P<0.05) at days 9 and 16. 5-Bromo-2-deoxyuridine labeled cel s existed in the wound in the treated groups at day 9. A significantly increased expression of endogenous keratinocyte growth factor was detected in the transplantation of Keratinocyte Growth Factor gene modified epidermal stem cel s group, and transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at day 16. A wel-advanced epithelialization was observed in transplantation of keratinocyte growth factor gene modified epidermal stem cel s plus low-frequency electromagnetic field exposure group at days 16 and 30. These results suggest that low-frequency electromagnetic fields enhanced wound healing fol owing the transplantation of keratinocyte growth factor gene modified epidermal stem cel s.

BACKGROUND:Inflammatory factor plays an important role in restenosis after bal oon injury. Sphingosine1-phosphate receptor 1 can enhance the expression of inflammatory factor and promote development and progression of this pathological process.
OBJECTIVE:To observe the expression of the inflammatory factors and sphingosine1-phosphate receptor 1 after bal oon injury of the rat carotid artery and effects of fingolimod hydrochloride on reducing inflammatory reaction.
METHODS:Sixty Sprague-Dawley rats were equal y and randomly divided into four groups. In the blank control group and negative control group, left common carotid artery was only isolated, and left external carotid artery was ligated. In the bal oon injury group and drug intervention group, rat models of carotid artery injury were
established by bal oon injury on the left common carotid artery. In the negative control and drug intervention groups, the rats were intraperitoneal y injected with fingolimod hydrochloride 1 mg/kg. In the blank control and bal oon injury groups, the rats were intraperitoneal y injected with an equal volume of saline. Samples were col ected at 3, 7 and 21 days.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that the proliferation of blood vessel was remarkable in the bal oon injury group, but attenuated in the drug intervention group. The appearance of blood vessels was normal in the blank control group and negative control group. Real-time fluorescent quantitative PCR revealed that cyclooxygenase 2 and prostaglandin E2 mRNA expression levels were significantly lower in the drug intervention group than those in the bal oon injury group at 7 days (P<0.05). Cyclooxygenase 2 and prostaglandin E2 mRNA expression levels were significantly higher in the bal oon injury group and drug intervention group than those in the blank control group and negative control group at the same time point (P<0.05). Western blot assay results revealed that sphingosine1-phosphate receptor 1 expression was high in early stage of injury, and then reduced in late stage of injury. In particular, protein expression further decreased after drug intervention. Results indicated that fingolimod hydrochloride suppressed inflammatory reaction of injured blood vessels and lessened the stenosis of injured blood vessels by regulating cyclooxygenase 2 and prostaglandin E2 mRNA expression using sphingosine1-phosphate receptor 1.

Objective To investigate the relationship between job burnout and psychological health diathesis of the senior medical workers in Tianjin.Methods 263 medical workers were tested by Psychological Health Diathesis Scale (PHDS) and Chinese Maslach Burnout Inventory (CMB[).Results (1) Total detection rate of burnout was 69.3％.Detection rate of mild burnout was 43.0％,moderate 20.2％,and severe 6.1％.(2) Explain rate of self-concept and emotionality to the job burnout was 31.5％.Conclusion Job burnout is prevalent in the senior medical workers in Tianjin.Emotionality,interpersonal diathesis and self-concept,to some extent,can forecast job burnout.

Objective To compare the efficacy and side effects between systemic chemotherapy and hepatic arterial infusion by combination of oxaliplatin and 5-fluorouracil (FOLFOX-6) with 5-fluorouracil in the patients who have developed hepatic metastasis after colorectal cancer operation. The factors that would affect the prognosis without operational treatment were also analyzed. Methods 46patients who had signed the informed consents were allocated into two groups: the group with general chemotherapy (Trial Group includes 26 cases) and the one with hepatic arterial infusion chemotherapy (Control Group includes 20 cases). The total effective rate, the prognosis, the cytoxicitic side effects,quality of life, the total survival rate and the responses were the main parameters determined. Kaplan-Meier was used to analyze Mono-factor to the prognostic responses and the Cox mode was used to analyze poly-factor to the prognostic responses. Results The overall survival rate was significantly higher by using systemic treatment versus HAI(median, 15. 0 v 11.2 months;P＜0.05). The difference in overall responsive rate (CR+PR) between the two groups was statistically significant (50% v 10%;P=0. 011). No significant difference was found in PS scale during the treatment. (P=0. 126). Except for myelosuppression and abdominal pain, no significant difference was found in the other side effects. Univariate analysis revealed that the invasive lesions to serosa, the distribution of liver metastases, the size and number of liver metastases, primary carcinoma involving lymph nodes and the treatment were correlated with prognoses. Cox regression analysis showed that the larger diameter of liver metastases, the number of liver lesions, primary carcinomas involved in serosal layer and the treatment modules were independent prognostic factors. Conclusions The oxaliplatin-based FOLFOX-6 chemotherapy regiment has a better responsive rate and survival rate than the traditional infusion with 5-fluorouracil to the main hepatic artery for interventional therapy. The diameter of the hepatic metastasis larger than 5em, multiple hepatic metastasis and the primary lesions penetrating serosal layer suggest the poor prognosis. The oxaliplatin-based systematic chemotherapy has a better prognosis. Therefore,it is worth carrying on further study on modification of traditional hepatic arterial infusion and on evaluation of therapy by combination of the hepatic arterial infusion with the systematic chemotherapy.

OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

Objective:To explore protective effect of atorvastatin on blood vessels in early stage of atherosclerosis (AS).Methods:A total of 120 patients without AS plaques,who had >2 cardiovascular risk factors and received control cardiovascular risk factors therapy,were randomly divided into four groups:control group (did not receive atorvastatin),atorvastatin 5mg group,10mg group and 20mg group (received corresponding dose of atorvastatin). All patients were followed up for six months,changes of thromboxane B2 (TXB2),6-Keto-prostaglandin F1α (6-Keto-PGF1α),brachial-ankle pulse wave velocity (baPWV),ankle brachial index (ABI)and intima-media thickness (IMT)were observed.Results:There were no significant changes in ABI and IMT between before and after treat-ment among four groups (P >0.05 all).Compared with baseline,TXB2、baPWV levels significantly rose,6-Keto-PGF1αlevel significantly decreased after treatment in control group and 5mg group;in contrast,TXB2、baPWV lev-els significantly decreased,6-Keto-PGF1αlevel significantly rose after treatment in 10mg group and 20mg group(P <0.05~ < 0.01).After treatment six-month,compared with control group and 5mg group,the TXB2 [(148.3 ± 29.2)pg/ml,(142.3±30.6)pg/ml vs.(111.5±22.8)pg/ml,(104.9 ± 17.4)pg/ml]、baPWV[(1621.1 ± 136.1) cm/s,(1597.7±125.3)cm/s vs.(1232.9±132.3)cm/s,(1178.2±155.1)cm/s]levels significantly decreased,6-Keto-PGF1α[(104.7±66.1)pg/ml,(102.2±70.3)pg/ml vs.(132.8±48.3)pg/ml,(139.1±66.3)pg/ml]level significantly rose(P <0.05~<0.01)in 10 mg group and 20 mg group.Conclusion:Atorvastatin has protective effect on blood vessels in early stage of atherosclerosis,and 10mg atorvastatin may be the minimum effective dosage to protect blood vessels.

HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.