BACKGROUND:Culture supernatant containing myocardiocyte has been demonstrated to induce differentiation of bone marrow-derived mesenchymal stem cel s into myocardiocyte-like cel s. This may associate with some or several cytokines in the culture supernatant.
OBJECTIVE:To explore if the supernatant of myocardiocyte induces bone marrow-derived mesenchymal stem cel s to differentiate into myocardiocyte-like cel s is associated with the different cytokine content in the supernatant of myocardiocyte.
METHODS:Bone marrow-derived mesenchymal stem cel s were isolated and cultured in vitro by the whole bone marrow adherent culture. Cardiocytes were isolated and cultured by enzyme digestion. Bone marrow-derived mesenchymal stem cel s (1×108/L) and cardiocytes (1×105/L) were cultured for 72 hours and the supernatant was col ected. Hepatocyte growth factor, insulin-like growth factor 1, platelet-derived growth factor, stem cel factor, fibroblast growth factor and vascular endothelial growth factor levels in culture supernatant of bone marrow-derived mesenchymal stem cel s and cardiocytes were detected by enzyme-linked immunosorbent assay.
RESULTS AND CONCLUSION:The content of insulin-like growth factor 1, platelet-derived growth factor and fibroblast growth factor in the supernatant of cardiocytes was significantly higher in cardiocytes group compared with bone marrow-derived mesenchymal stem cel s group (P<0.01). Results indicated that insulin-like growth factor 1, platelet-derived growth factor and fibroblast growth factor in the supernatant of cardiocytes may have capability to induce bone marrow-derived mesenchymal stem cel s to differentiate into myocardiocyte-like cel s, and insulin-like growth factor 1 may serve as the main cytokine.

Seven adult albino rats were used for this study. The 50％ HRP solution was injected into the cervical enlargment unilaterally. Labelled neurons were mainly found in the nucleus paraventricularis, nucleus lateralis, nucleus paraventricularis, nucleus lateralis, nucleus perifornicalis, nucleus hypothalamicus anterior, nucleus hypothalamus posterior, nucleus suprachiasmaticus, nucleus supraopticus and nucleus periventricularis ipsilaterally. Most of the labelled neurons were found in nucleus paraventricularis hypothalamicus, less in the nucleus lateralis and only a few in other nuclei.The authors suggest that the efferent projections from the hypothalamus to the spinal cord in the rat may play a role in the integrative function of the spinal cord. which may be involved in the process of acupuncture analgesia.

Attempts were made to determine the cells of origin from the thalamus to the spinal cord by using the retrograde tracer technique with horseradish peroxidase (HRP). The solution of HRP was injected into the intumescentia cervicalis unilaterally in eight albino rats. The results indicated that the HRP-labeled cells were located in the parafascicular nucleus, subparafascicular nucleus and the ventromedial nucleus of the thalamus. The function of spinal projection from these neuclei were discussed.

The efferent projections to the spinal cord from the brain stem were studied with HRP. Fifty per cent solution of HRP was slowly injected into the intumescentia cervicalis at six points on its right side in 10 albino rats and the HRP-labelled cells were found in the following nuclei:1. Of the midbrain: the nucleus linearis, nucleus raphe dorsalis, substantia nigra, nucleus ruber, nucleus cuneiformis, nucleus colliculus superior, nucleus Edinger-Westphal, nucleus Darkschewitsch, nucleus interstitialis (Cajal), nucleus ventralis tegmenti and nucleus dorsalis tegmentalis;2. Of the ports: the nucleus raphe magnus, nucleus medianus raphe, nucleus trigemini principalis, nucleus lemnisci lateralis, nucleus reticularis pontis oralis, nucleus reticularis pontis caudalis, locus ceruleus, nucleus subceruleus, nucleus vestibularis lateralis, nucleus vestibularis medialis, nucleus parabrachialis and nucleus olivaris su- perior;3. Of the medulla oblongata: the nucleus raphe obscurus, nucleus raphe pallidus, nucleus gracilis, nucleus cuneatus, nucleus tractus spinalis nervi trigemini, nucleus tractus solitarius, nucleus commissuralis (Cajal), nucleus reticularis lateralis, nucleus reticularis paramedianus, nucleus reticularis gigantocellularis and nucleus olivaris inferior;4. Of the cerebellum: the nucleus medialis and lateralis.The functions of spinal projection from the nuclei raphe, reticularis, locus ceruleus, substantia nigra and the nucleus dorsalis tegmentalis have been discussed.

Background and purpose:Bladder cancer radical surgery is dififcult with many perioperative complications, and the learning curve is long. To introduce the clinical application of complete retrograde radical cystectomy and consequent abdominal cavity reconstruction in male bladder cancer, and to provide a standardized surgical procedure with minimum perioperative complications and short learning curve. Methods:From Apr. 2012 to Apr. 2013, 110 cases of male patients with bladder cancer received this procedure in our department, with a median age of 64 (35-83) years;Preoperative characters, surgical parameters, perioperative complications, pathology, long-term complications and short-term prognosis were collected and analyzed. Results:The median number of lymph nodes resected in operation was 12 (8-16);Neurovascular bundles were reserved bilaterally in 65 cases, and unilaterally in 31 cases;The complete procedure including urinary diversion took 4.4 (2.2-6.0) hours, with a median time of opened abdominal cavity of 43.0 (5.0-75.0) minutes;The median blood loss was 140.0 (50.0-600.0) mL, and 4 patients needed transfusion; Median time of abdominal and pelvic drainage was 10.0 (6.0-15.0) days, the median gastrointestinal recovery time was 2.5 (1.0-12.0) days, and the median postoperative hospital stay was 17.0 (10.0-39.0) days;Grade 2 Clavien-Dindo classiifcation (CDC) of surgical complications that required medical intervention were found in 19 cases, CDC grade 3 or above were found in 8 cases;Mild to moderate postoperative ileus happened in 5 cases, all recovered in median 2 (1-4) weeks with supportive treatments;There were no perioperative deaths. All samples were sent to pathological analyses. After a median follow-up of 9 (3-15) months, no complications of or above CDC grade 3 happened, and there were no recurrence. Conclusion:Complete retrograde radical cystectomy in male bladder cancer provided clear anatomical approach, reliable neurovascular bundle preservation, less blood loss, limited abdominal organs disturbance and better surgical exposure; With respect to tumor control, more peritoneal was retained for subsequent abdominal cavity reconstruction. The introduced procedure effectively speeded up gastrointestinal recovery, reduced postoperative complications, especially the incidence of ileus and its severity, and shortened hospital stay. The learning curve of this procedure for urologists was short, and further investigation was warranted.

Objective:To artificially design and express a recombinant protein named as ScFv-pLLO by fusing ScFv gene of Rituximab(C2B8)and dominant antigen epitopes from listeriolysin O(LLO),and studying its anti-tumor activity.Methods:VH and VL gene sequences of C2B8 against CD20 were acquired by searching United States Patent database,and ScFv sequence was constructed by linking VL and VH with a short peptide linker.Two CD4+T cell epitopes from LLO were selected and designed to splice ScFv sequence.The recombinant gene of ScFv-pLLO was cloned into prokaryotic expression vector and purified after induction.The capacity of ScFv-pLLO target-binding to B-cell lymphomas was evaluated by flow cytometry ( FCM ) and co-immunoprecipitation ( Co-IP ) .The effects of ScFv-pLLO on B-cell lymphomas proliferation and apoptosis were detected respectively.The immunogenicity of ScFv-pLLO was assessed by lymphocyte proliferation assay.Results: ScFv-pLLO was successfully expressed.It could bind to different B-cell lymphomas cell lines and obviously inhibit the growth of Raji cells as well as inducing apoptosis.Moreover,ScFv-pLLO was able to stimulate proliferation of spleen lymphocytes of immunized mice.Conclusion: The recombinant protein ScFv-pLLO can target-bind to B-cell lymphomas,and perform inhibitory effect and induce apoptosis on Raji cells that indicate ScFv-pLLO retain the capacity of ScFv derived from monoclonal antibody against CD20.Besides, ScFv-pLLO can induce immune response.This study provides a basis for further research about the role of ScFv-pLLO on simulating tumor cell antigens as well as being tumor vaccine adjuvant.

Objective To construct a nomogram for estimating disease free survival of node-positive penile cancer treated with surgery.MethodsThe clinicopathological data of 32 node-positive penile squamous cell carcinoma patients were retrospectively reviewed.All patients had undergone primary tumor excision and regional lymphadenectomy.The pathological records showed that 10 were N1,13 were N2 and 9 were N3.Median level of preoperative squamous cell carcinoma antigen (SCCAg) was O.9 μg/L.With a median follow-up of 16.5 months,13 patients developed recurrence.Based on the stage,histological grade and preoperative serum squamous cell carcinoma antigen level,a nomogram was drawn on the basis of the Cox regression model.Results Stage and preoperative serum SCCAg levels were independent prognostic factors for disease free survival in node-positive penile cancer treated with surgery.The hazard ratio of SCCAg level and N stage was 2.76 (95％ CI:1.40-5.44,P=0.0034) and 28.51 (95％ CI:2.55 -319.11,P =0.0066).The nomogram demonstarted good discrimination and calibration with a concordance index of 0.855.Conclusions Based on the N stage and preoperative serum SCCAg level,a nomogram to predict disease free survival in node-positive penile cancer treated with surgery was set up.The prediction model could be helpful in predicting the risk of recurrence.

Objective To evaluate the recurrence patterns and possible prognostic factors in primary penile cancer treated with penectomy and standard lymphadenectomy.Methods The records of 73 patients with penile squamous cell carcinoma who underwent penile amputation and regional lymphadenectomy between 1990 and 2005 were reviewed.The median follow up was 32 ( 16 - 183) months.Molecular markers including p53 and Ki-67 were detected using immunohistochemistry.Recurrence patterns,pathological characteristics,and patient outcomes were analyzed.ResultsTwenty( 27.4％ ) patients developed recurrences at a mean of 11 (6 - 17 ) months postoperatively and 3 cases remained alive at last follow-up.The mean cancer specific survival of the recurrent patients was 26 ( 16 - 83 ) months.Distant metastasis and multiple recurrences were common among patients with high-grade tumor (P =0.017 ).Univariate analysis revealed that higher primary tumor stage,higher pathological grade,lymph node metastasis,extra-nodal involvement and positive p53 protein expression were significantly associated with decreased 3-year disease free survival rates.Cox regression identified that only pathological grade ( P =0.025 ) and lymph node status ( P =0.024 ) were independent predictors of disease free survival.ConclusionsPathological grade and lymph node status were independent predictors of disease free survival in patients with penile squamous cell carcinoma in this cohort.Patients with high-grade disease are likely to recur in a distant or multiple pattern.

Objective To explore the accuracy and practical significance of the 3-dimensional (3D) reconstruction of the liver and the abdominal blood vessels based on the data of 64-slice spiral computerized tomography (64S-SCT). Methods The 2D images of the liver and the abdominal blood vessels were collected after TLC-scanning with 64S-SCT. The 3D images of the liver, hepatic internal duct system and the abdominal blood vessels were reconstructed by the medical image processing system. The volume of the 3D reconstructed liver was compared with that of the actual liver measured by the 64S-SCT, and the portal vein of the reconstructed liver model was compared with that reconstructed by the Mxview workstation based on the 64S-SCT data. Results The 3D models of the liver, hepatic internal duct system and abdominal blood vessels were vivid, and could be revolved and observed at any direction. The spatial relationship among main intrahepatic ducts was clearly displayed. The actual liver volume and the condition of the intrahepatic ducts were accurately demonstrated. The reconstructed liver, the branches of the hepatic artery and portal vein, and the abdominal blood vessels were simultaneously displayed by adjusting the transparency of the liver model. The portal vein of the reconstructed liver model completely matched that reconstructed by the Mxview workstation based on the 64S-SCT data. Conclusions The 3D models of the liver, hepatic internal duct system and abdominal blood vessels reflect the human anatomy structure, and provide reliable virtual organs and vessel system for virtual liver surgery.

Objective To evaluate the prognostic value of bilateral inguinal lymph node metastases in node-positive penile cancer. Methods Sixty patients with surgically resected node-positive penile cancer were analyzed. All the patients underwent regional lymph node dissection. Recurrence free survival curves were plotted by Kaplan-Meier method and compared by the Log-rank test. Multivariate survival analysis was performed using Cox regression model. Results Of all the patients, 18 cases had bilateral inguinal lymph node metastases. The 3-year recurrence free survival rate in patients with bilateral invovlement was significantly lower than those with unilateral disease (26.7 % vs 65.3 %, χ2 =10.6, P=0.001). In order to evaluate wether the prognostic significance of bilateral lymph node metastases was dependent of increased number of positive nodes, the two predictors were included in multivariate survival analysis.Both the number of metastatic nodes and bilateral nodal disease were independent prognostic factors (P ＜0.05).Comparisons of survival curves showed those patients with bilateral nodal disease and more than 2 metastatic nodes had worst outcome. Conclusion Bilateral inguinal lymph node metastases is an important prognostic factor in penile cancer irrespective of the number of positive lymph node.