ObjectiveTo investigate the expression and the clinical significance of diacylglycerol kinase α (DGKα) and protein kinase C (PKC) in human hepatocellular carcinoma (HCC).MethodsDGKα and PKC expressions in the samples from 60 pathologically confirmed HCC patients were analyzed by immunohistochemistry. The relationship between DGKo expression and clinical pathology factors was analyzed.ResultsThe expression positive rates of DGKo and PKC were highest in normal liver tissues [90.0％ (9/10) and 100.0％ (10/10)].The positive rates were 81.7 ％ (49/60) and 71.7 ％ (43/60) in HCC tissues,respectively,and were 58.3 ％ (35/60) and 61.7 ％ (37/60) in carcinoma adjacent tissues,respectively.In three liver tissues,the positive rates of DGKα and PKC were significantly different (P ＜0.05).The location of both kinases in hepatocytes translocated from cytoplasm/nucleus to membrane.The expressions of DGKα and PKC were positively correlation(r =0.495, P ＜ 0.05), The DGKα expression was correlated to differentiation type,portal venous tumor thrombus and TNM staging(all P ＜ 0.05).ConclusionDGKa is expressed with high activity in advanced stage and poorly differentiated HCC. It may be promote the pathological process of HCC.

Objective To explore the mechanism underlying the rapid shortening of outflow tract and the formation of the right ventricle of the embryonic mouse heart .Methods Serial sections of embryonic mouse hearts from embryonic day 9 (E9) to E12(3 to 5 embryos for each stage)were stained with antibodies against α-sarcomeric actin (SCA), α-smooth muscle actin (SMA), GATA-4, myosin heavy chain (MHC), proliferating cell nuclear antigen (PCNA) or active caspase-3 (CAS-3).Results At E11, the aortic sac and the distal border of cardiac outflow tract had regressed towards the ventricle into the pericardial cavity , while GATA-4、SCA and SMA staining showed that precursors from the second heart field were differentiating into cardiomyocytes adding to the arterial pole of the heart to lengthen the outflow tract .The length of outflow tract rapidly shortened at E12.Before and during its shortening , no CAS-3 positive cell was detected in the entire outflow tract.During E10-12, the cardiomyocytes in the right ventricle and proximal outflow tract wall proliferated inward to form trabeculae, with some trabeculae extending into the ridges .Proximal extremities of the outflow tract ridges were gradually myocardialized remodeling into the trabeullar right ventricle wall .At E12, scattered SCA and SMA staining cells and SCA and SMA weak positive mesenchymal cell clusters , which were continuous with the outflow tract myocardium were detected in the mesenchymal proximal outflow tract ridges .These results suggested that the proximal outflow tract was remodeled into the right ventricle by trabecularization , during which mesenchymal ridges were trabecularlly myocardialized . Conclusion Ventricularization of the proximal outflow tract contributes to the trabecular right ventricle and resultes in the vapid shortening of outflow tract in the mouse embryonic heart .Cardiomyocyte appoptosis and transdifferentiation are found to play a more limited contribution during this process .

Objective To investigate the early development of the sinus venosus and the cardiac conduction system (CCS) of human embryonic hearts. Methods Serial transverse sections of 29 human embryonic hearts from Carnegie stage 10 to Carnegie stage 16 (C10-C16) were stained immunohistochemically with antibodies against α-smooth muscle actin(α-SMA),α-sarcomeric actin(α-SCA) and desmin ( DES ). Results During C12 and C13, the sinus venosus formed by confluence of systematic veins at the caudal end of the pericardial cavity could be recognized in the mesenchyme of primitive transverse septum. The mesenchymal cells of the sinus venosus gradually differentiated into α-SCA positive cardiocyocytes. At C14, the sinus venosus was within the pericardial cavity due to expansion of the pericardial cavity and incorporated into the right atrium. Differentiation of DES positive conductive cardiomyocyte was initiated in the right wall of atrio-ventricular canal of C10 embryonic heart and with the development, extended towards the myocardium of the interventricular sulcus to form His bundle, left and right bundle branches as well as the ventricular trabecular myocardium. In the atium, the strong expression of DES was first detected in the dorsal wall of C11 atrium. At C13, unique myocardial band showing α-SCA, α-SMA and DES expression in the left dorsal wall of the sinus venosus were found to be continuous with the basal wall of left atium and the dorsal wall of the atrio-ventricular canal, this band might be related to the development of conduction system from sinoatrial node to atrio-ventricular canal. During C14 to C16, primary conduction pathway of atria with strong DES expression was formed that extended from sinoatrial node along venous valves, DES positive myocardium in the dorsal and ventral walls of the atria to the right atrio-ventricular canal, respectively. Conclusions The mesenchyme of the primitive transverse septum is the heart forming field of human embryos responsible for formation of sinus venosus myocardium, cardiomyocytes are differentiated from mesenchymal cells in the primitive transverse septum and progressively added to the venous pole of the heart tube to form myocardial sinus venosus. The differentiation of CCS of the early human embryo initiates in the atrio-ventricular canal and develops gradually towards the arterial and venous poles of the heart tube. By C16, DES positive embryonic CCS can be clearly recognized morphologically.

Objective To investigate the expression and distribution of Diacylglycerol Kinase α (DGKα) mRNA in human hepatocellular carcinoma (HCC),and to explore the function of DGKα in the metastasis of hepatocellular carcinoma.Methods Tissues from 30 cases of HCC and 5 normal liver tissues were collected immediately after surgical resection.Semi-quantitative RT-PCR and in situ hybridization were used to detect the expression levels and distribution of DGKα mRNA,respectively.Results Semi-quantitative RT-PCR showed that the expression level of DGKα mRNA in HCC (0.798±0.317) and normal tissues (0.908±0.425) was significantly higher than those in carcinoma adjacent tissue with cirrhosis (0.205±0.102,P<0.05).In situ hybridization demonstrated that the number of DGKα mRNA positive hepatocytes in HCC [(57.6±6.3)/mm2] and normal tissues [69.8±8.7)/mm2] was significantly higher than those in carcinoma adjacent tissue with cirrhosis [(26.3±4.9)/mm2,P<0.05]; DGKα mRNA was expressed in the cytoplasm of hepatocytes in HCC and carcinoma adjacent tissue with cirrhosis,and in the nuclear of hepatocytes in normal tissues.Conclusion The present study suggests that DGKa may play important roles in carcinogenesis and progressing of HCC.

Objective To investigate the origin of α-SMA positive cells in the outflow tract ridge of the embyonic mouse heart and to explore the ultrastructure change of the mesenchymal cells during the ridges fusion. Methods Sections of embryonic day 10(E10d) to E14d mouse embryonic hearts were stained by immunohistochemistry assay with monoclonal antibodies against α-smooth muscle actin (α-SMA), α-sarcomeric actin(α-SCA) and in situ hybridization method with PlexinA2 probe. The outflow tract ridges fusion was observed by transmission electron microscopy at E12.5d. Results From E10d to E11d, PlexinA2 positive cells were seen in the neural tube with mesenchymes around it, the aortic sac and aortic arch. These cells also migrated into the outflow tract ridge along the aortic sac wall and part of them expressed α-SMA. At E12d, PlexinA2 was expressed in the spinal ganglia, the pharyngeal mesenchyme, the aorto-pulmonary septum and the ascending aorta and pulmonary trunk. The septum showed α-SMA strongly positive. But only a few of α-SMA positive cells were observed in the ascending aorta and pulmonary trunk. At E12.5d, two clusters of condensed mesenchymal cells in the outflow tract ridges formed and began to express PlexinA2 weakly and α-SMA strongly. When the ridges began to fuse, the endothelial cells formed a cellular seam, which rapidly broke into pieces and disappeared and were replaced by the sparsed mesenchymal cells containing a few of microfilaments. Two clusters of condensed mesenchymal cells gradully moved to merge. It was noted that some mesenchymal cells contained plenty of microfilament bundles, mitochondria and focal contacts between them. Some mesenchymal cells only had a few of microfilaments and plasma membrane fusion was seen between them. Conclusionα-SMA positive cells in the outflow tract cushion may be derived from cardiac neural crest. The endothelial cells might participate in the fusion of the outflow tract ridges by endothelial-mesenchymal transformation. Mesenchymal cells of the condensed cell mass contain plenty of microfilament bundles and focal contacts or membrane fusion, which contribute to the ridges fusion.

Objective To investigate the effects of duration of surgery on flash-induced visual evoked potentials (VEP) in patients undergoing spinal surgery in prone position.Methods Eighty-two ASA Ⅰ or Ⅱ patients of both sexes aged 20-76 yr weighing 43-96 kg undergoing spinal surgery in prone position were divided into 3 groups according to the duration of surgery:group S≤2 h ( n =34) ; group M 2-4 h ( n =38) and group L≥4 h ( n =10).VEP was monitored using protektor VEP monitoring device (Xltek Co.,Canada).The latency,amplitude and recovery time of wave P100 were recorded before and 10 min after induction of anesthesia and at the end of surgery.Results Compared with group S,the amplitude of wave P1000 was significantly decreased at the end of surgery in group M,the lantency of wave P100 was significantly prolonged,while the amplitude of wave P100 was decreased at the end of surgery in group L ( P ＜ 0.05).Compared with group M,the lantency of wave P100 was significantly prolonged,while the amplitude of wave P100 was decreased at the end of surgery in group L ( P ＜ 0.05).Compared with groups S and M,the recovery time of wave P100 was significantly prolonged in group L ( P ＜0.05).There was no significant difference in the recovery time of wave P100 between groups S and M ( P ＞ 0.05).Conclusion Duration of surgery (≥4 h) can affect flash-induced VEP,the longer the duration,the stronger the effects.

The integrated curriculums of basic medicine in Shanxi Medical University are as follows: nine basic medical courses were integrated into 10 medical modules, with additional training including PBL and TBL case discussion, clinical clerkship, flipped classroom and other non-integrated subjects etc. In order to assess the interest of students for integrated curriculum, their intrinsic motivation, as well as their comprehension and application of medical knowledge, we gave anonymous questionnaire to 149 students, 16 teachers in basic medicine, and 10 teachers in clinical medicine. Results showed that more than 90% students were willing to take the integrated curriculum and participate in PBL and TBL case discussion, and students who were unwilling to take the curriculum were less than 10%, they thought that the knowledge of new curriculum system was incoherent. The proportion of students from higher grade who were unwilling to participate in the flipped classroom was increased from 6.7% to 95.0%. Most of the teachers both in basic and clinic medicine believed that new curriculum system was helpful for students to comprehend basic medical knowledge and strengthen their discriminating ability, but did not function in improving students' practical ability. In the further reform on teaching, details like the coherence of knowledge and the content selected when self-studying should be improved.