Cultivating clinical comprehensive ability is one of the core and main purposes for surgical clinical education.According to the characteristics in clinical teaching of hepatobiliary surgery,we combined basic theory with clinical practice and discussed on the problems of how to improve medical students' clinical comprehensive abilities including clinical operational ability,logical thinking ability,scientific research innovative ability and the doctor-patient communication ability and how to cultivate medical ethics and professional competence from aspects of basic clinical skills training,translational medicine concept,etc.

Objective To study the inhibition effect of leech extract on HepG2 cells. Methods Human hepatocellular cancer cell line HepG2 were treated with different concentrations of leech extracts which were extracted by method of freeze-thawing with liquid nitrogen and contrasted with that by method of water extracting and ethanol precipitating. The inhibition effects and cell morphous were examined by MTT assay and Acridine orange (AO) fluorescent staining method respectively. Result The 6～15 mg/mL drug concentrations of leech extract by method of freeze-thawing with liquid nitrogen had an obvious inhibition on proliferation of HepG2 cells in a dose-dependent manner, and the effect was better than that by method of water extracting and ethanol precipitating (P

Objective To study the role of Vibrio vulnificus cytolysin(rVvhA) induced THP-1 apoptosis and calcium influx.Methods CCK-8 cell proliferation kit,Fluo3/AM staining and AnnexinV/PI staining were performed to identify the apoptosis and calcium influx induced by rVvhA in THP-1 cells.Results rVvhA could induce THP-1 apoptosis and up-regulate the cellular calcium concentration.BAPTAAM could enhance the calcium influx induced by rVvhA in THP-1.Conclusion rVvhA had cytotoxic to THP-1 cells by inducing apoptosis and triggering extracellular calcium influx.

It has been found by Ju('81)that HRP could be anterogradely transported forlong distances.Based on this fact the rate of anterograde transport was investigatedin the present study,and,at the same time,the rate of retrograde transport and thetime of disappearance of HRP from the labeled sites were observed.Twenty tworabbits,weighing 2～2.2 kg,were used.The HRP was injected into the lower lumbarspinal cord unilaterally.The animals were sacrificed after 12 hours to 7 days andwere processed with benzidine and o-dianisidine.The anterogade labeling in the dorsalaccessory olivary nucleus and the retrograde labeling in the red nucleus and thenucleus raphe pallidus were chosen for study.It was quite unexpected to find that the anterograde and retrograde labelingsparalled each other in time as well as in intensity.They appeared at the same timeand waxed and waned practically in full accord.Labeling of the brainstem firstappeared at 18 hour's survival,reached its peak at the 2nd day,and then graduallyfaded away till at the 7th day when only traces of labeling remained.In the casewhich showed the earliest labeling the distance between the injection site and theobex,which was at about the same level as the most densely labeled parts of thedorsal accessory olivary nucleus and nucleus raphe pallidus,was measured to be 265mm and the rate of transport,the same for anterograde and retrograde transports,was calculated to be more than 350mm per day.Counting from the time of theearliest labeling,HRP was found to remain at the labeled site for as long as 5～6 days.The period of net accumulation of HRP at the labeled sites was about 1 day.Thus,the best survival period would be roughly the time required for HRP to travel thefiber tract plus 1 day.Neurons of the red nucleus and the nucleus raphe pallidus differ greatly intheir morphology,physiology as well as biochemistry.The results in them however,were similar.This,in connection with observations on other nuclei,both anterogr-adely and retrogradely labeled,suggests that the rate of transport and the duration ofpreservation of HRP in the central nervous system are basically the same in differentneuronal systems.

HRP was injected into the right side of the cervical or lumbar enlargement ormidthoracic segment of adult cats.Retrogradely labeled cells in the red nucleuswere examined and their somatotopic localization was determined.1.The Rubro-spinal projection was mainly crossed with obvious somatotopiclocalization.The dorsal medial part of the red nucleus projected to the cervicalenlargement.The ventral lateral part of the red nucleus projected to the lumbarenlargement.The neurons between these two parts projected to the thoracic cord.2.In cervical and lumbar injected cases a few labeled cells were found also inthe ipsilateral red nucleus,suggesting a minor ipsilateral rubrospinal projection exist-ed.No ipsilateral labeling was found in the thoracic injection cases.3.The rubrospinal tract could at least project to as low as L 6 segment of thespinal cord.

The somatotopical projection from the lateral vestibular nucleus to differentlevels of the spinal cord in the cat was studied with the horseradish peroxidasemethod.It was found that in the lateral vestibular nucleus the cells projecting tothe cervical and lumbar segments of the cord were distributed over the entire ros-trocaudal extent,each with an area of concentration.The caudal and middle thirdsof the nucleus,especially its dorsolateral part,projected to the lumbar cord.Thecells projecting to the cervical cord were concentrated at the rostral-middle thirds ofthe nucleus and those to the thoracic cord were concentrated at the dorsal part ofthe caudal-middle thirds.The area projecting to the thoracic levels seemed tocoincide with that projecting to the lumbar levels.The cells projecting to thelumbar cord were largest in number,those to the cervical cord less and those to thethoracic cord least.

Objective To investigate the molecular and cell signal transduction mechanisms by Lactobacillus cell wall extract(LCWE)inducing human-β-defensin-2(hBD-2)expression in human vaginal epithelial cells.Methods The induction of hBD-2 in human vaginal epithelial cells(WZV-1)by LCWE was observed using real-time PCR and Western blot.After stimulating WZV-1.the activation of NF-κB and p38MAPK signaling pathways were determined by Western blot.The induction of hBD-2 in WZV-1 cells by LCWE was observed with signaling pathways inhibitors of NF-κB and p38MAPK using real-time PCR and Western blot.Results The results showed that LCWE significantly upregulated hBD-2 expression in the time and dose-dependent manner.The maximal stimulatory effect of LCWE on the expression of hBD-2mRNA in WZV-1 cells were observed at the concentration of 50μg/ml after treatment for 8 h.After stimulation by 50μg/ml LCWE,Western blot analysis demonstrated that the phosphorylation of p38MAPK increased at 0.5 h significantly,peaked at 1 h,moreover the concentration of NF-κB in nucleus increased at 0.5 h significantly(P<0.05),peaked at 2 h.Blocking with inhibitor of NF-κB and(or)p38MAPK pathways results in decreased levels of HBD-2 expression.Conclusion These findings suggest that p38MAPK and NF-κB pathways play the important roles in induction of hBD-2 expression by LCWE in human vagihal epithelial cells.

HRP was injected into the cervical (3 cases) or lumbar (2 cases) spinal cord unilaterally in 5 adult cats. Labeled cells were discovered in the hypoth alamus and nearby areas. There was no obvious difference in labeling between cervical and lumbar injection cases. Labeled cells were found bilaterally with ipsilateral preponderance.he paraventricular nucleus was most heavily labeled; the posterior and lateral hypothalamic areas were less. A few labeled cells were found in the dorsal hypothalamic area and the supramamillary nucleus. Forel's area was also weakly labeled and occasional cells were found in the subthalamic nucleus and zona incerta.We were unable to find labeled cells in the dorsomedial nucleus. Labeling of the supramamillary nucleus, which was found in this sutdy, has not been mentioned in the literature available to us.

The projection from the lower brain stem to the spinal cord was studied with HRP method by injecting HRP into the cervical and lumbar enlargements of the spinal cord of 10 cats.The follwing nuclei were found to have spinal projections:1) The reticular formation: Numerous retrogradely labeled cells were seen in the nuclei of gigantocellularis and medullae oblongatae centralis subnucleus ventralis, fewer in the nuclei pontis centralis caudalis and oralis. A few labeled neurons were also found in the nuclei of medullae oblongatae centralis subnucleus dorsalis, parvocellularis, paragigantocellularis laterlaris, paramedium reticularis subnucleus ventralis and cuneiformis. Most of them projected to the spinal gray matter ventral to the dorsal horn except the nuclei of medullae oblongatae centralis subnucleus dorsalis and parvocellularis which projected mainly to the dorsal horn.2) The raphe nuclei: Nuclei of raphe pallidus, magnus and obscurus projected to both the cervical and lumbar enlargements, while nucleus raphe dorsalis only to the gray matter ventral to the dorsal horn of the cervical enlargement.3) The gracile and medial cuneatus nuclei projected somatotopically to ipsilateral spinal cord.4) The cranial nuclei: The Edinger-Westphal nucleus, nuclei of nervi oculomotor principalis, tractus spinalis nervi trigemini and tractus solitarii and the nuclei vestibularis lateralis, medialis, superior and spinalis were found to project to spinal cord. There was somatotopic arrangement in the nucleus vestibularis lateralis.5) The nuclei of locus coeruleus, subcoeruleus, parabrachialis lateralis and medialis, neucleus retroambigualis and the Kolliker-Fuse nucleus projected bilaterally to the spinal cord.6) The red nucleus: Large amount of labeled cells were seen in the controlateral red nucleus. The ventrolateral part of the red nucleus projected to the cervical enlargement while the dorsornedial part to the lumbar enlargement.7) The superior colliculus and the griseum centrale mesencephali projected to the gray matter ventral to the dorsal horn of the cervical enlargement.

HRP was injected into C_6, C_7, L_5, L_6 or T_(5～7) spinal gray. The retrogradely labeled cells and anterogradly labeled terminal arborizations were traced in L_5 and T_5 in cervical injection cases, C_5 and T_5 in lumbar cases, and C_5, L_5 in thoracic cases.Large amount of labeled cell were consistently found in laminae Ⅶ (including the lateral horn of the thoracic cord) and Ⅷ, next numerous in lamina Ⅴ and Ⅹ region. Lamina Ⅰ was found labeled in the cervical and lumbar cord, but virtually not found in the thoracic cord. Labeled cells were few and inconsistent in laminae Ⅳ and Ⅵ. A few labeled lamina Ⅸ cells could be found in the thoracic cord, only occasionally in the lumbar cord, but could not be found in the cervical cord. Laminae Ⅱ and Ⅲ were free from labeled cells in all cases. The labeled terminal arborizations were more widespread and could be found in practically every lamina, being densest in region Ⅹ, laminae Ⅶ (including the lateral horn of the thoracic cord) and Ⅷ. In lumbar injection cases a prominent plexus of labeled terminal arborization could be seen in the ventrolateral part of the C_5 ventral horn. Corresponding plexus was not found in the lumbar cord in cervical injection cases.Some labeled terminal arborizations were found in close approximation to or overlapping with labeled cells or unlabeled motor neurons.These results imply that: (1) The occurance of labeled cells and labeled terminal arborizations in the lateral part of lamina Ⅶ (lateral horn) suggests that intersegmental connection is related to visceral activities as well as somatic ones. (2) Labeled cells were consistently found in region Ⅹ which was also the area of dense labeling of terminal arborization, suggesting that this region plays a remarkable role in intersegmental connection. (3) Long ascending bundle connects the lumbar enlargement with the ventrolateral part of the motor neurons directly. (4) The close approximation of labeled terminal arborization and labeled cells signifies direct intersegmental feedback circuit.