ObjectiveTo discuss the effects of apoptosis and invasion of RBE cells caused by miRNA 21 suppression and further investigate the potential role of miRNA-21 plays on target mRNA regulation. MethodsThe RNAi technology was employed to suppress the expression of RBE cells.The changes in RECK mRNA and protein expressions were detected by RT-PCR and Western blotting respectively. Changes occurred in apoptosis was closely monitored by flow cytometry (FCM). The invasion of RBE cells was analyzed in vitro by invasion assay (transwell). ResultsThe expression of miRNA-21 was clearly suppressed while the RECK mRNA and protein were over-expressed. The rate of apoptosis was significantly accelerated and there was a dramatic decrease in RBE cells' ability to invade after miRNA-21 knockdown. ConclusionThrough miRNA-21 suppression, the rate of apoptosis of RBE cells was accelerated whereas their invasion ability was greatly reduced. RECK was found to be the target gene of miRNA-21 which participates in the regulation process of regulation.

Antiviral Agents ; pharmacology ; Humans ; Interferons ; pharmacology ; Pharmacology, Clinical

Objective: To investigate the effect and mechanism of helix B surface peptide (HBSP) on myocardial ischemia reperfusion injury (MIRI) in experimental mice.
Methods: The MIRI model was established by ligation of anterior descending coronary artery of the mice for 45 min and followed by corresponding treatment at 5 min before reperfusion. A total of 64 male ICR mice were randomly assigned to 4 group:①Sham group,②MIRI group, the mice received normal saline at 5 min before reperfusion,③HBSP group, MIRI mice received HBSP at 5 min before reperfusion and④HBSP+PD98059 group, MIRI mice received PD98059 (a speciifc blocker of ERK1/2) at 20 min before reperfusion and followed by HBSP at 5 min before reperfusion.n=16 in each group, all animals were treated for 2 hours. The area of myocardial infarction (MI) was detected by TTC-EB double staining method, the myocardial apoptosis rate was examined by TUNEL method, the levels of protein expression of ERK1/2 and phosphorylation of ERK1/2 were measured by Western blot analysis.
Results: Compared with MIRI group, HBSP group presented decreased MI area, decreased myocardial apoptosis rate and increased phopsphorylation level of ERK1/2, allP<0.05. Compared with HBSP group, HBSP+PD98059 group showed decreased phopsphorylation level of ERK1/2, increased myocardial apoptosis rate and increased MI area, allP<0.05.
Conclusion: HBSP may reduce the MI area via inhibiting myocardial apoptosis and therefore, protecting the experimental mice from MIRI; the mechanism might be related to the activation of ERK1/2 pathway.

Objective To investigate CT signs of peripheral small cell lung cancer (SCLC).Methods The CT signs of 78 patients with SCLC confirmed by pathology were retrospectively reviewed.According to the presence of mediastinal lymph node metastasis and its size, 78 cases of peripheral SCLC were divided into two types: typeⅠ(isolated lesion) and typeⅡ(lung lesion + lymph nodes).Type Ⅱwere divided into two subtypes:type Ⅱa (short diameter of lymph nodes of pulmonary hilar and mediastinum less than 10 mm) and type Ⅱ b (short diameter of lymph nodes of pulmonary hilar and mediastinum greater than or equal to 10 mm).Results Of the 78 SCLCs, typeⅠwas 7 cases, and typeⅡwas 71 cases,including 8 cases of typeⅡa and 63 cases of typeⅡb.All of the lesions were soild density.The shape were round or oval in 52 cases, vermicular or spindlein 9 cases, and other shapes in 17 cases.Among 71 cases performed CT enhancement, there were 9 cases with homogeneous enhancement, 58 cases with heterogeneous enhancement, 4 cases with non-enhancement large necrosis area.These cases showed the following CT signs: smooth edge in 65 cases, coarse edge in 12 cases, blurred edge in 1 case;air bronchogram in 3 cases, vacuole sign in 4 cases, calcification in 4 cases;lobulation sign in 46 cases, spiculated sign in 5 cases;thickening of the bronchovascular bundle in 41 cases, pleural indentation in 6 cases, marginal ground-glass opacity in 5 cases, vascular convergence sign in 1 case;emphysema in 42 cases;obstructive pneumonia in 4 cases;bronchus abruptly interruption on the edge of the nodules in 18 cases;enlargement of mediastinal lymph nodes in 63 cases, the diameter of mediastinal lymph nodes larger than the primary lesions in 42 cases;and a little pleural effusion in 9 cases.Conclusion Solid density, smooth margin with lobulation,and significantly enlarged mediastinal lymph nodes are common signs in peripheral SCLC.Thickening of the bronchovascular bundle indicates reletively advanced stage.

OBJECTIVETo transfect a short hairpin RNA (shRNA) against survivin gene into human T lymphoblastic leukemia cell line Jurkat, and to explore the effects on apoptosis and proliferation of transfected cells.

METHODSThe survivin-shRNA expression vector were constructed and transfected into Jurkat cells. Expression of survivin mRNA and protein were assessed by RT-PCR and Western blot analysis respectively. Apoptosis index of transfected Jurkat cells was quantified by flow cytometry. The potential of cell proliferation was described by cell growth curves.

RESULTSIn survivin-shRNA transfected Jurkat cells, survivin mRNA levels were significantly reduced by 66.67% ( transient transfection) and 60.69% ( stable transfection) respectively, compared with that in control-shRNA treated group and PBS treated group (P < 0.05); and the levels of survivin protein were significantly reduced by 63.41% (transient transfection) and 60.18% (stable transfection), compared with that in the two control groups (P < 0.05). Apoptosis index was significantly increased during both transient and stable transfection, respectively [(22. 41 +/- 2.83)% and (20.73 +/- 2.56)% (P < 0.05)]. Survivin-shRNA also inhibited the proliferation of Jurkat cells.

CONCLUSIONSVector-based survivin-shRNA can effectively reduce the expression of survivin gene, induce apoptosis

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Expression ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; Jurkat Cells ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; RNA, Small Interfering ; pharmacology

BACKGROUND: Preliminary studies have found that poly(lactic-co-glycolic acid) (PLGA) can improve the compressive strength and degradability of calcium phosphate cement. OBJECTIVE: To prepare a self-setting calcium phosphate cement which has better mechanical properties, biocompatibility and degradability on the basis of the previous findings. METHODS: Beta-tricalcium phosphate (β-TCP), pure calcium phosphate cement (CPC) and PLGA powder were mixed at different mixing ratios for preparation of PLGA/β-TCP/CPC. Setting time, compressive strength, elastic modulus and degradation properties of the composite bone cement were evaluated to screen the optimal level of β-TCP. MC3T3-E1 cells were cultured in CPC extract (control), PLGA/β-TCP/CPC extract (experimental), α-MEM containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody (negative control), and 6.4% phenol liquid (positive control). MTT method was used to detect cell proliferation at 1, 3, 5 days after culture, and lactate dehydrogenase activity in culture media was detected at 1 and 3 days after culture. MC3T3-E1 cells were seeded on the surface of PLGA/β-TCP/CPC and pure CPC respectively, and were then observed by scanning electron microscopy after 3 days. RESULTS AND CONCLUSION: Initial setting time and final setting time among of the composite bone cement were increased with the increasing of β-TCP content, but had no significant difference from those of the CPC (P ＞ 0.05). The compressive strength and elastic modulus of the composite bone cement were higher than those of the CPC, and moreover, the composite bone cement with 20% β-TCP exhibited the highest compressive strength and higher elastic modulus as compared with the other groups. Therefore, PLGA/20% β-TCP/CPC was selected in the cell test. Moreover, the degradation properties of the composite bone cement were also better than those of the CPC. (3) With the growth of culture time, cell absorbance value and lactate dehydrogenase activity were gradually increased in the experimental group, and no difference existed between the experimental group and the negative control group. The cells in the experimental group also grew well. (4) MC3T3-E1 cells grew well and fully extended on the surface of PLGA/β-TCP/CPC, and cell pseudopodia on the material surface were tightly adhered to the material. To conclude, PLGA/20% β-TCP/CPC has better compressive strength, elastic modulus, degradation properties and cytocompatibility relative to the CPC, and moreover, the composite bone cement has no obvious cytotoxicity.

OBJECTIVE: To establish standardized methods and parameters of the isolated heart coronary angiography through the experiment of in vitro porcine heart by MSCT. METHODS: Based on different perfusion volume (50, 60 and 70 mL) and different perfusion-imaging time (5, 10 and 20 min), the in vitro porcine coronary artery was injected liposoluble and water-soluble contrast agents using remodel angiography equipment and scanned by MSCT. And the 3D image results were compared. The images were recorded and evaluated by 2 radiologists and analyzed by statistical software. RESULTS: Liposoluble contrast agent affected the images by damaging and infiltrating the fats around the coronary artery, while the water-soluble contrast agent didn't affect the images. The groups with 60 mL or 70 mL perfusion and 5 min perfusion-imaging time had the best images. CONCLUSION The suitable parameters of the angiography lay the foundation of postmortem coronary angiography.
Animals ; Coronary Angiography/veterinary* ; Coronary Vessels/diagnostic imaging* ; Heart ; Imaging, Three-Dimensional/methods* ; In Vitro Techniques ; Multidetector Computed Tomography/veterinary* ; Predictive Value of Tests ; Sensitivity and Specificity ; Software ; Software Validation ; Swine

OBJECTIVETo report the first case of non-composite combined liver and intestinal allotransplantation in China. The technical aspects of the case and pros and cons of such an approach versus composite technique were discussed.

METHODSThe patient suffered from short bowel syndrome and TPN-related liver damage. A non-composite technique was used in this case. During operation, the whole 380 cm intestine was transplanted with systemic drainage and aortic inflow, while the liver graft was placed in a piggyback fashion. Warm ischemic time of donor graft was 2 min and 30 seconds, and cold ischemic duration for intestinal and liver graft was 6 hours and 40 and 8 hours and 7 utes respectively. Postoperative immunosuppression management includes tacrolimus, methylprednisolone, MMF and Zenapax.

RESULTSThe recipient recovered smoothly with no evidence of rejection on days' follow up. Now he is maintained well on enteral nutrition.

CONCLUSIONNon-composite technique should be considered in adult recipients, especially those with a history of abdominal infections or multiple laparotomies.

Adult ; Humans ; Intestines ; transplantation ; Liver Transplantation ; Male ; Short Bowel Syndrome ; therapy ; Transplantation, Homologous ; methods ; Treatment Outcome

OBJECTIVETo study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.

METHODSEukaryotic expression plasmid pcDNA3. 1-antisense (As) survivin was constructed and transfected into Jurkat T lymphoblastic lymphoma cell lines with high expression survivin mRNA by use of lipofectmine gene transfer technique. Expression of survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical and Western blot. The effect of transfecting survivin antisense mRNA on the growth of Jurkat cell lines was monitored by population doubling time (PDT) and Apoptotic indexes (AI). The morphologic features were observed in transfected cells by light and electric microscopes. MTT assay was used to analyze the response of transfected cells to CTX and MTX.

RESULTSCompared with the control cells, the expression of survivin mRNA and protein were reduced after transfected pcDNA3. 1-Assurvivin 48 h, 5 w and 6 w, PDT (52 h) was prolonged. Apoptotic indexes were higher in transfected antisense survivin mRNA cells [20.2% (48 h)], 6.2% (5 w) and 6.8% (6 w) than control ones [2.1%, 1.3% (48 h)] and [1.3% (5 w) and 1.0% (6 w)]. The cells grow slowly and the dead cells increase and some swelling and apoptotic cells were observed in transfected pcDNA3. 1-Assurvivin groups by invert, light and electric microscopes. The Jurkat cell line of transfected pcDNA3. 1-Assurvivin had higher sensitivity to CTX and MTX. The rate of inhibition was higher in transfected group. There is a significant difference between the transfected group and untransfected one, P < 0.05.

CONCLUSIONSThe result indicated that survivin gene was very important for growth of Jurkat cells. To inhibit the expression of survivin will be significant in therapy of T lymphoblastic lymphoma. Survivin gene might be a target of therapy.

Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents, Alkylating ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cyclophosphamide ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Jurkat Cells ; cytology ; metabolism ; K562 Cells ; cytology ; metabolism ; Lymphoma ; pathology ; Methotrexate ; pharmacology ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Plasmids ; RNA, Antisense ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

OBJECTIVETo explorer the effectiveness of enriched bone marrow stem cells technique for lumbar fusion.

METHODSWith the randomization and control principles, 2 graft materials [Enrichment bone marrow mesenchymal stem cells hybridized with beta-tri calcium phosphate (composite graft group), autologous iliac crest bone graft (autograft group)] were compared in posterior lumbar fusion procedures. 56 patients with degenerative disc disease, lumbar instability or spinal stenosis, were included. The volume of cells suspension in pre- and post-enrichment and the number of nucleated cells (NCs) were identified. The number of osteoprogenitor cells was estimated by counting the colony-forming units which express alkaline phosphatase (CFUs/ALP+). Then the efficiency of the enrichment was evaluated. Clinical follow-up with roentgenogram and Oswestry scale scores was performed for outcome evaluation.

RESULTS(249 +/- 31) ml bone marrow per patient from bilateral iliac crests was aspirated peri-operatively. About (43 +/- 11) ml enriched bone marrow was collected. The number of NCs was concentrated from (15.9 +/- 3.3) x 10(6)/ml to (44.1 +/- 10.8) x 10(6)/ml, CFUs/ALP+ was significantly increased from (118 +/- 86)/ml to(486 +/- 305)/ml. The follow-up was about (26.3 +/- 7.5) months. There was no significant differences in age, gender, disease and fusion segments between the two groups. The fusion rate was 93.3% and 96.2% for composite graft group and autograft group, respectively (chi2 = 0.2146, P = 0.6432). There was no difference in operation time between the two group (t = 0.5243, P = 0.6022), but blood loss in composite graft group was more than that in autograft group (t = 6.4664, P < 0.01). Cell salvage for auto-transfusion could transfuse back half of the blood loss during operation. No hematoma or chronic soreness in the bone marrow donor sites of composite graft group occurred, but a little exudation or moderate swelling in the wound happened in 4 cases which disappeared under medical treatment. Meanwhile, 15.4% patients had hematoma in the iliac bone donor site and 26.9% patients had chronic soreness, but no case had wound problem in autograft group. As for Oswestry scale scores, there was no significant difference between the two groups.

CONCLUSIONSThe enrichment technique of autologous bone marrow stem cells can greatly increase the concentration of MSCs. It is a rapid and safe method used peri-operatively. The composite material of enriched MSCs and porous beta-TCP is a good bone substitute in posterior spinal fusion.

Adult ; Aged ; Bone Substitutes ; Bone Transplantation ; methods ; Calcium Phosphates ; Female ; Follow-Up Studies ; Humans ; Ilium ; transplantation ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; instrumentation ; methods ; Stem Cell Transplantation ; Transplantation, Autologous ; Treatment Outcome