Objective To evaluate the effect of laparoscopic ultrasonography in laparoscopic cholecystectomy. Methods Laparoscopic ultrasonography system of Sharplan Company was used for the examination of the liver, gallbladder, bile duct and pancreas in 410 patients performed by laparoscopic cholecystectomy. Results The rates of biliary duct abnormality, preoperative diagnosis change, preoperative inexpectant disease finding, bile injury and residual stone were 3.5%, 4.4%, 2.4% and 0%, respectively. Conclusion During laparoscopic cholecystectomy, laparoscopic ultrasonography is helpful for the improvement of correct diagnosis and the decrease of complications.

Objective To compare and evaluate dose at optic structures (eyeballs, optic nerves and chiasma) between conventional and CT simulated treatment planning for nasopharyngeal carcinoma. Methods Thirty three patients with T1 to T4 nasopharyngeal carcinoma underwent simulated spiral CT with picker PQ 5000 and AcQ plan 4.1.1 software system. At first irradiation portals were delinated by the simulation films taken on the conventional X ray simulator using the digital reconstruction (DRR) of the treatment planning system (TPS) supplemented by the patient's CT (or MRI) and clinical findings. Then, the gross tumor volume (GTV),clinical target volume (CTV) and planned target volume (PTV) were delineated on every section of the CT scans. At the same time, the silhouette of the nearby important organs was drawn out ,paying special attention to the backward passage of the retrobulbar optic nerves up to the optic chiasma. The final portals were designed by the 3 dimensional relation between the tumor and its surrounding organs. Then dose distribution was separately calculated by two independent TPSs,with 100% at the target center. Finally,the maximum dose (D max ),average dose (D ave ) and median dose (D med ) of the tumor target area, eye ball,optic nerve and also the length of optic nerve included within the portal of these two technics were compared.Results The dose distribution in nasopharyngeal GTV and CTV were satisfactory both in CT simulated and conventional planning. However, the D max ,D ave and D med all showed significant differences. Those by CT simulated planning were obviously lower than those by conventional planning. CT simulated planning was able to spare more optic nerve by leaving it out of the irradiation portal,thereby,avoiding unwanted radiation.Conclusion CT simulated treatment planning is able to reduce the superfluous radiation to the optic nerve in external irradiation of nasophargngeal carcinoma.

Objective To observe the morphologic changes of of vascular buds in vertebral cartilage endplate in age-specific rabbits and also to investigate the correlation between the changes of vascular buds and interverbral disc degeneration. Methods There were 15 New Zealand white rabbits in our study,which include three groups, 2-week-old rabbits, 1-year-old rabbits and 3-year-old rabbits, and each groups had five rabbits. The X-ray radiograph, histology and scanning electron microscope were used to observe the changes of vertebral cartilage endplate. According to Miyamoto standard, the interverbral disc was graded 1-5, and scored 1-5 respectively. Results The changes of micro-vascular structure of vertebral cartilage endplate were observed during aging. Under the scanning electron microscope, the vascular structure degenerated gradually, and disappeared in the end. The blood vessels in the central region of the vertebral cartilage endplate reduced more obviously than those in periphery region. The severe degeneration was found in vertebral endplate, compared with intervertebral disc. The changes of vascular buds in rabbits vertebral cartilage endplate had positive correlation with the vertebral endplate calcification and the interbertebral disc degeneration. Conclusion Changes of vascular buds in vertebral endplate may accelerate intervertebral disc degeneration.

BACKGROUND:Renal tubular-interstitial lesion and fibrosis induced by peritubular capil ary reduction is a common pathway for various chronic kidney diseases which eventual y develop into end-stage renal failure. How to increase the density of peritubular capil ary network is the key to resolving renal interstitial fibrosis. Netrin-1, as a potent mitogen of vascular endothelial cel s, can promote the migration and proliferation of vascular endothelial cel s and induce angiogenesis. OBJECTIVE:To observe the protective effects of naked netrin-1 plasmid transfer on the remnant renal function of 5/6 nephrectomized rats and the effects of naked netrin-1 plasmid transfer on peritubular capil ary network, and to further discuss the possible mechanism. METHODS:Thirty Sprague-Dawley rats were randomly divided into sham, model and treatment groups. Upper and lower one-third of the rat left kidney was resected in the model and treatment groups, and then the right kidney was resected after 1 week to prepare remnant kidney models in rats. IRES2-EGFP and pCMV6-XL5-Netrin-1-IRES2-EGFP pGenesil-NPs plasmids were intravenously injected into the left renal vein while resecting the right kidney of rats in the model and treatment groups, respectively. RESULTS AND CONCLUSION:Compared with model group, the levels of blood urea nitrogen and serum creatinine decreased, the degree of renal interstitial fibrosis al eviated, the density of peritubular capil aries increased, and the netrin-1 protein expression in renal tubular cytoplasm increased in the treatment group. These findings suggest that the naked netrin-1 plasmid transfer can significantly improve the renal function of the 5/6 nephrectomized rats, reduce the pathological lesion and renal interstitial fibrosis of the remnant kidney, increase the density of peritubular capil aries, and decrease the expression of hypoxia-inducible factor 1α, thereby improving the anoxic condition of renal interstitial tubules.

Objective To investigate the effect of combining propofol with hydrogen on organ damage and inflammation of sepsis in cecal ligation and puncture (CLP) mice model.Methods One hundred and forty male C57BL/6 mice were randomly divided into groups (n = 28): sham group, CLP group, propofol group, H2 group, and propofol and H2 group. The sepsis was induced by CLP operation. Mice in sham group did the same operation with ligation and puncture. The mice of propofol group and propofol and H2 group were given 50 mg/kg propofol through tail vein at 1 hour and 6 hours after CLP and the mice of H2 group and propofol and H2 group were given 5 mL/kg H2-rich saline i.p. at 1 hour and 6 hours after CLP. The survival rates were observed during 7 days in twenty mice of each group. Inferior vena cava blood and part lung, liver and kidney tissue were collected for detection of the concentration of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and high mobility group box 1 (HMGB1) at 24 hours after CLP in the 40 animals left (eachn = 8). Then, the rest tissue of lung, liver and kidney tissue were harvested to test histopathology and histological score.Results The 1, 2, 3, 5, 7 days survival rate of septic mice were 80%, 40%, 20%, 10%, and 0%, respectively. The survival rate of animals increased significantly after propofol or hydrogen-rich treatment, and the combined treatment can further increase survival rate to 90%, 75%, 60%, 55%, and 55%, respectively. Compared with the sham group, inflammatory factors were significantly increased in blood and organ tissues, cell degeneration, necrosis, congestion and inflammatory cell infiltration in lung, liver and kidney, and tissues histological scores were significantly increased. The levels of inflammatory factors were reduced in blood and tissues, cell degeneration, necrosis, congestion and inflammatory cell infiltration were alleviated in lung, liver and kidney, and tissues histological scores were decreased after propofol or hydrogen-rich treatment compared with CLP group; these indicators were further improved in propofol and H2 group compared with propofol group or H2 group [2, 3, 5, 7-day survival rate: 75% vs. 60%, 65%; 60% vs. 50%, 50%; 55% vs. 45%, 40%; 55% vs. 40%, 40%; blood TNF-α (ng/L): 367±74 vs. 612±132, 588±117; blood IL-1β (ng/L): 321±68 vs. 502±95, 476±86; blood HMGB1 (μg/L): 4.6±0.9 vs. 7.0±1.4, 6.8±1.3; lung TNF-α(ng/g): 307±70 vs. 512±132, 488±102; lung IL-1β (ng/g): 367±77 vs. 571±108, 466±89; lung HMGB1 (μg/g):5.1±1.0 vs. 7.8±1.7, 7.1±1.5; liver TNF-α (ng/g): 247±57 vs. 431±112, 389±87; liver IL-1β (ng/g): 267±58 vs. 417±85, 399±76; liver HMGB1 (μg/g): 4.2±1.1 vs. 7.1±1.6, 6.6±1.2; kidney TNF-α (ng/g): 257±41 vs. 480±89, 448±82; kidney IL-1β (ng/g): 258±39 vs. 409±68, 411±66; kidney HMGB1 (μg/g): 3.9±0.7 vs. 6.8±1.2, 5.7±1.0; histological scores: lung: 1.22±0.28 vs. 2.61±0.49, 2.58±0.44; liver: 1.38±0.32 vs. 2.76±0.51, 2.62±0.46; kidney: 1.19±0.25 vs. 2.43±0.41, 2.36±0.40; allP < 0.05].Conclusions Both propofol and H2 can improve the survival rate of sepsis, reduce tissue damage and the release of cytokines, and combined application of the two treatment was better.

Objective To investigate the usefulness of acoustic radiation force impulse (ARFI) elastography for noninvasive evaluation of liver fibrosis in rats.Methods A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury,and 10 saline-injected rats were used as normal control.Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight.Several rats in the group with DNM injected and the normal control group were randomly selected and sacrificed at each of the following post-injection time:day 5,7,10,14,21,24,and 28.And their livers were taken for pathology analysis.All the rats underwent ARFI elastography before sacrificed in order to acquire a shear wave velocity (Vs) to represent liver stiffness.Correlation between Vs and the histological finding was analysed.ResultsAmong 58 successfully modeled rats,9,13,14 and 12 rats were found to be with S1,S2,S3 and S4 of liver fibrosis pathologically,respectively.And 10 rats were found to be with severe inflammatory activity without any fibrosis.Values of Vs increased with the stage of liver fibrosis ( P ＜0.05).There was a significant correlation between Vs and stage of liver fibrosis ( r =0.947,P =0.000).The areas under ROC curve for the diagnosis of fibrosis S≥S1,S≥S2,S≥S3 and S=S4 were 0.983,0.995,0.999 and 0.964,respectively;for the cutoff values of Vs were 1.59 m/s,2.13 m/s,2.33 m/s and 2.51 m/s,respectively,the sensitivity was 95.8％,92.3％,100％ and 84.6％,and specificity was 100％,100％,96.9％ and 95.6％,respectively.The values of Vs in the group with severe inflammatory activity were significantly higher than those in the control group ( P =0.000).ConclusionsARFI has a relatively high value in the evaluation of liver fibrfosis in rats,while severe inflammatory activity may affect its accuracy.

BACKGROUND:To improve the mechanical properties and uncontrolability of degradation of decelularized matrix, wemanufactured genipin cross-linked decelularized annulus fibrosus matrix/chitosan hydrogels as annulus fibrosus tissue-engineereds caffold.
OBJECTIVE:To observe the biocompati bility of annulus fibrosus-derived stem cels with genipin cross-linked decelularized annulus fibrosus matrix/chitosan hydrogels, and the degradation of hydrogels in vivo.
METHODS:Genipin cross-linked decelularized annulus fibrosus matrix/chitosan hydrogels caffoldswere prepared, and the internal structure and hydrophilic property of hydrogels was detected by scanning electron microscopy and contact angle meter, respectively. Afterwards, passage 1 rabbit annulus fibrosus-derived stem cels were seeded on thescaffoldsfor 3 days, cel morphology was observed by inverted immun of luorescence microscopy and scanning electron microscopy after cytoskeleton staining, andadditionaly, curve of cel growth was obtained. In the meanwhile, thescafflodswere transplanted into theintermuscular space of New Zealand white rabbits, and the degradation and inflammatory reaction of the hydrogels were observedafter4 weeks.
RESULTS AND CONCLUSION:The prepared hydrogelsc affoldshowed porous and network structure and the contact angle was (39.94±1.61)°; annulus fibrosus-derived stem cels adhered wel on thescaffold surfacewithafaster growth rate. Furthermore, at 4 weeks after trans plantation, the hydrogelin vivowas degraded, and inflammatory cels appeared. These findings indicate that genipin cross-linked decelularized annulus fibrosus matrix/chitosan hydrogels have good biocompati bility.

Objective To evaluate the safety of dynamic anterior plate fixation for cervical distractive flexion injuries and compare its rigidity between different types of plate design. Methods Twelve sets of cadaveric calf spine were used in this test. All the specimens were made into distractive flexion injury models (C4-C5) ac- cording to Allen's method. After discectomy and grafting, they were randomized into three groups in which Orion, Codman, and Window instrumentations were used respectively. The stiffness of each construct was tested in flexion, extension, lateral bending and axial torsion conditions sequentially. Results Compared with an intact cervical spine, the range of motion (ROM) of an injured cervical spine increased whatever plate was applied. Orion in- strumentation presented stiffness the closest to that of the normal control, except for less torsional stiffness. Codman instrumentation provided stiffness close to that for normal and Orion groups only in lateral bending. Window's was the weakest mad not enough in all kinds of movement. Conclusions Static anterior fixation is the first choice for cervical injuries. Dynamic plate fixation may sacrifice stiffness to some extent, especially when a shifting kind of design is to be chosen.

BACKGROUND: Endothelial progenitor calls are the cells that can form new blood vessels in the way of angiogenesis in the body,which updates the conventional theory of angiogenesis, vascular damage and repair after birth and provides new ideas for research and treatment of ischemic diseases.OBJECTIVE: To investigate the effects of dog umbilical cord blood endothelial progenitor call (UCB-EPC) transplantation on angiogenesis after myocardial infarction.DESIGN, TIME AND SETTING: An in vivo cytological experiment was performed at the Laboratory Center of Xinhua Hospital between May 2006 and March 2007.MATERIALS: One full-term pregnant hybrid dog was included for preparation of UCB-EPCs. Thirty-six adult dogs were randomly divided into a cell transplantation group (n = 18) and a model control group (n = 18).METHODS: Acute myocardial infarction model was established in each group by ligation of antedor descending coronary artery.In the cell transplantation group, 2 mL physiological saline containing 5×10<'6> BrdU-labeled EPCs was injected into the coronary artery, while in the model control group, simple physiological saline of the same amount was given. At 1,4, and 8 weeks after transplantation, dogs were sacrificed for harvesting myocardial tissue.MAIN OUTCOME MEASURES: Myocardial infarction was confirmed by hematoxylin-eosin staining. Myocardial angiogenesis was observed by BrdU immunohistochemical staining. The number of infarcted myocardial vessels was calculated by yon Willebrand (vW) factor staining.RESULTS: There was plenty of scar tissue, flbroblasts, and small vessels in the myocardial infarction region. In the cell transplantation group, brown yellow particles (BrdU-positive expression) appeared in some nuclei in small vessels from infarcted myocardium. Newly formed vessels were not found in the model control group. In the cell transplantation group, brown yellow particles (vW factor-positive expression) appeared in the cytoplasm of the vascular endothelial cells in the myocardial ischemia and infarction regions, vW factors were not expressed in the model control group. At 1, 4, and 8 weeks after myocardial infarction,there was no significant difference in vessel counts no matter in myocardial ischemia region or in myocardial infarction region between the call transplantation and model control groups.CONCLUSION: EPCs derived from UCB of pregnant dog can participate in the formation of blood vessels but can not promote angiogenesis after acute myocardial infarction.

BACKGROUND:Endplate cartilage degeneration initiates intervertebral disc degeneration. AMP-activated protein kinase (AMPK) regulates the formation and degradation of cartilage. OBJECTIVE:To explore the role of AMPK in an in vitro natural degeneration model of chondrocytes derived from endplate of rat intervertebral discs. METHODS:Morphology of in vitro subcultured endplate chondrocytes of rat intervertebral discs at passages 0, 2, and 5 were observed under an inverted microscope fol owing cytoskeleton staining. Chondrocyte phenotype, proliferation, and the cartilage marker genes (type II col agen, proteoglycan, SOX-9, matrix metal oproteinase-3 and-13), and AMPK phosphorylation were determined by toluidine blue staining, MTT assay, real-time PCR analysis, and western blot assay, respectively. RESULTS AND CONCLUSION:The altered morphology, decreased proliferation ability, and phenotype loss were observed in chondrocytes with increased passage number. Gene expression of type II col agen, proteoglycan, SOX-9 was significantly decreased;while gene expression of matrix metal oproteinase-3 and-13 was significantly increased in endplate chondrocytes at passage 5 compared with those at passages 0 and 2. AMPK phosphorylation in endplate chondrocytes at passage 5 was significantly decreased. These findings indicate that AMPK phosphorylation is involved in in vitro natural degeneration of chondrocytes derived from the endplate of rat intervertebral discs, and the degeneration of endplate chondrocytes and intervertebral discs can be inhibited through the regulation of AMPK activity.