We isolated polyA~+ mRNA direcdy from tumor tissue of ovarian carcinoma using Oligotex~(TM) Direct mRNA Kit (QIAGEN) to synthesize the first and second strand cDNA. The ds-cDNA termini were blunted with pfu DNA poly-rnerase. The blunted cDNAs were added EcoR I adaptor, and then were digested by Xho I . Small cDNA molecules(

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.

Objective To evaluate biocompatibility of Allomax mesh implanted in different planes of abdominal wall in a rats model.Methods SD rats were randomly assigned to the profacial group (Onlay group),the retro-rectus group (Sublay group) versus the intraperitoneal group(IPOM group) according to different abdominal wall planes the mesh implanted,Adhesion and shrinkage of the mesh were observed,and quantitative measurements were conducted in fibroblast ingrowth,scaffold degradation,extracellular matrix deposition and numbers of vascular ingrowth after 1,3 and 6 months mesh was implanted.Results Macroscopic observation showed both Onlay and Sublay groups was superior to IPOM group in abdominal wall integration,which included shrinkage,relocation and adhesion of the mesh at all the time points,and most or whole of the mesh had incorporated with host abdominal wall at 6 months.Most of the mesh had not incorporated with host abdominal wall and shrinkage and relocation of the mesh were found in IPOM group at 6 months.Microscopic investigation showed lipocytes appeared in the mesh in Sublay group at 3 months,and numbers of ingrowth of fibroblast and neovascularization in Sublay group were significantly less than in Onlay and IPOM group at 6 months.Scaffold degradation and extracellular matrix deposition were remarkably less in Sublay group in comparison with Onlay group and IPOM group after 1,3 and 6 months.Conclusions Biocompatibility of AlloMax mesh implanted in profacial plane of abdominal wall was superior to implanted in retro-rectus plane and intraperitoneal plane as showed in a rat model.

Objective To explore the value of glypican-3(GPC-3)mRNA and paternally expressed 10(PEG10)mRNA in peripheral blood in diagnosis of metastasis in hepatocellular carcinoma(HCC). Methods With SYBR Green I as fluorescence signal,real-time fluorescent quantitative reverse transcription-polymerase chain reaction was employed to detect the expression of GPC-3 mRNA and PEG10 mRNA in peripheral blood from patients with HCC with metastasis(n=8),HCC without metastasis(n=12)and hepatic cirrhosis(n=11),and receiver operator characteristics curve(ROC)and specific parameters were adopted to analyse their value in predictive and exclusive diagnosis. Results The expression of GPC-3 mRNA and PEG10 mRNA in HCC with metastasis was significantly higher than that in HCC without metastasis and in hepatic cirrhosis(P<0.05),while there was no significant difference in the expression of GPC-3 mRNA and PEG10 mRNA between HCC without metastasis and hepatic cirrhosis.In single test,the sensitivities in the differential diagnosis between HCC with metastasis and HCC without metastasis were 66.7%for GPC-3 mRNA and 72.2%for PEG10 mRNA,and the specificities were 91.7%and 91.7%.respectively.The areas under ROC were 0.748 for GPC-3 mRNA and 0.812 for PEG10 mRNA.With two markers in parallel test,the sensitivity,specificity,negative likelihood and diagnostic accuracy were 90.7%,84.O%,0.11 and 83.3%,respectively.In serial test,the sensitivity,specificity,positive likelihood and diagnostic accuracy were 60.5%,98.7%,45.5 and 73.3%,respectively. Conclusion Detection of GPC-3 mRNA and PEG10 mRNA in peripheral blood may help to predict blood metastasis and extrahepatic metastasis of HCC,and PEG10 mRNA works better than GPC-3 mRNA.The serial test of GPC-3 mRNA and PEG10 mRNA is helpful to the predictive diagnosis of peripheral blood metastasis of HCC.

BACKGROUND:The use of cemented or uncemented hemiarthroplasty for unstable intertrochanteric fractures in the elderly remains controversial.Therefore,it is necessary to conduct a comparative study on the effectiveness and safety of these two methods.OBJECTIVE:To compare the clinical efficacy of cemented and uncemented hemiarthroplasty for unstable intertrochanteric fractures in the elderly.METHODS:Clinical data of 93 elderly patients with unstable intertrochanteric fractures in Department of Orthopedics,Shandong Energy Zaozhuang Mining Group General Hospital from May 2009 to May 2014 were analyzed retrospectively.All patients were divided into cemented (cemented bipolar hemiarthroplasty for fractures,n=54) and uncemented (uncemented bipolar hemiarthroplasty for fractures,n=39) groups.RESULTS AND CONCLUSION:(1) The amounts of postoperative drainage and blood transfusion in the cemented group were significantly less than those in the uncemented group (P ＜ 0.05),but the operation time was significantly longer (P ＜ 0.05).(2) There were no significant differences in the intraoperative blood loss,ambulation time,hospitalization time,postoperative complications,hip function,and mortality at 3 and 12 months postoperatively between two groups (P ＞ 0.05).(3) Postoperative X-ray showed that all patients had good prosthesis position.There were 10 patients (3 cases in the cemented group,7 cases in the uncemented group) with postoperative prosthesis subsidence,but all of them were less than 3 mm.(4) None of patients had heterotopic ossification,osteolysis around the prosthesis and acetabular cartilage wear during follow-up.Additionally,25 patients (16 cases in the cement group,9 cases in the uncement group) died during follow-up,without prosthesis loosening,and the remaining 68 patients were followed up for 2-7 years,(4.5±2.3) years on average,and none needed revision because of prosthesis loosening.(5) These results indicate that for senile unstable intertrochanteric fractures,both cemented and uncemented hemiarthroplasty can achieve satisfactory curative effectiveness and exhibits good safety.Notably,the cemented prosthesis has advantages in reducing postoperative drainage volume and blood transfusion.

Objective:To explore whether apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin can be cross-presented by dendritic cells(DCs) and enhance immune responses.Methods:DCs were induced from peripheral blood monocytes cells by GM-CSF/IL-4 for 6 d,then they were stimulated with either apoptotic ovarian cancer HO8910 cells,frozen-thawed HO8910 cells or control cells for 4 h.Their surface markers and phagocytotic ability were detected by flow cytometry and confocal microscopic scanning assay,respectively.DCs of different groups were cultured with CD8+ T cells isolated by magnetic cell sorting,and the ability of DCs to activate CD8+ T cells was evaluated by 3H-TdR,the activity of CTL to kill tumor cells was evaluated by LDH.Production of IFN-? by CD8+ T cells was measured by ELISPOT.Results:Apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin could be phagocytized by DCs,which subsequently promoted the maturation and antigen presenting ability of DCs.Apoptotic ovarian cancer cells implused DCs significantly promoted proliferation of CD8+ T cells compared with that of control cells(P

Objective:As the function of ??T cells and NK in the immunological therapy is showing more and more important,the characteristics of ?? T cells,NK and LAK were analyzed comparatively after that they were richened by isolating and incubating in vitro.Methods:The cells were collected using MACS after the cells were panned respectively with special monoclonal antibodies.Then the characteristics including proliferation,phenotype,cytotoxin and the down regulation blocked by specific antibodies were analyzed.Results:The ?? T cells can expand 600～800 times after culturing 2 weeks and the percent of CD3,CD8 and ?? expressed on the collected cells were 72.29%,58.02% and 65.98% respectively ?? T cells showed the high cytotoxin to K562(NK sensitive cell line),Raji and XG 7 cell lines(NK non sensitive cell lines).The percentage of cytotoxin reached 35.98%,52.27% and 69.08% respectively compared with ??T respectively.No obviously change of percent of ??T cytotxic ability to these target cells were observed using special MHC class I monoclonal antibody to block the ??T cell before coculturing the target cells with ?? T cells.Conclusion:All of ?? T cells,NK and LAK showed the non specific cytotoxin to tumor cells.The cytotoxic capability of ?? T cells did not be effected after blocking with MHC class I monoclonal antibody.These results demonstrated that ?? T cell could kill more kinds of tumor cells than NK and LAK.