Objective To investigate the effects of morphine dependence and withdrawal on neurosteroids and amino acid transmitters of rat amygdala. Methods Morphine dependence was induced by pretreatment with increasing doses of morphine for 7 days. Withdrawal was precipitated by naloxone (2mg/kg). Withdrawal syndromes were observed and scored. After decapitation, amygdala was dissected out. Nomadic and conjugated neurosteroids were extracted using liquid-liquid extraction and solid phase extraction. Concentrations of neurosteroids including dehydroepiandrosterone (DHEA), pregnenolone (PREG), allopregnanolone (AP), dehydroepiandrosterone sulfate (DHEAS) and pregnenolone sulfate (PREGS) were detected with HPLC-MS. Concentrations of glycine (GLY), glutamate (GLU) and gamma-aminobutyric acid (GABA) were quantitated by HPLC-ECD with pre-column OPA derivatization. Results Compared with saline control, the DHEA level in rat amygdala of morphine dependent group decreased by 33% (P＜0.01). Compared with naloxone control, the PREG and AP levels in rat amygdala of morphine withdrawal group increased by 45% (P＜0.05) and 42% (P＜0.05) respectively; the GABA level decreased by 18% (P＜0.01). Compared with morphine dependent group, the PREG and PREGS levels in rat amygdala of morphine withdrawal group increased by 60% and 40% respectively (P＜0.05); the glycine level decreased by 14% (P＜0.05). Conclusion The DHEA in rat amygdala may play a role in the development of morphine dependence but not involved in the manifestation of withdrawal symptoms. Other neurosteroids (including PREG, AP and PREGS) in rat amygdala seem to be involved in withdrawal but not in dependence. The synthesis and release of inhibitory amino acids in amygdala were depressed when withdrawal was precipitated by naloxone. The results suggest that different changes of neurosteroids and amino acids exist in stages of morphine dependence and withdrawal.

Objective To investigate the effects of electro-acupuncture on gastric emptying and Fos expression in the neuron of vagal-solitary complex (VSC) of rat bulbus after lipopolysaccharide (LPS) intraperitoneal injection (i.p.). Methods 40 male SD rats were randomly assigned to 4 groups: control group, LPS i.p. group, LPS i.p. plus electro-acupuncture at Tsusanli point group and LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint group, 10 rats for each group. Immunohistochemical techniques were used to detect the Fos expression in VSC. The animal's gastric emptying was measured by phenol red method. Results The rats with gastric emptying decreased greatly to (20.7±4.5)% 2.5 hours after LPS injection, and Fos-positive neurons were significantly found in VSC (83.2±6.6) compared with control group. However, in the group of LPS i.p. plus electro-acupuncture at Tsusanli point, the gastric emptying was up-regulated to (44.1±6.2)%, and the expression of Fos -positive neurons were down-regulated to (37.9±3.8) compared with LPS i.p. group. No significant difference was found between the group of LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint and the group of LPS i.p. Conclusion LPS i.p. can retard the gastric motility in rats, electro-acupuncture at Tsusanli point may well regulate the function in LPS model rats. This function may be connected with its protective effects on Fos immunoreactive neurons activity in VSC of rat bulbus.

Objective To study the role the pathogenesis of early acute lung injury (ALI) of rabbits induced by intravascular injection of endotoxin (ET) with the intervening method of Chloroquine. Methods Rabbits were randomly assigned to three groups: control group, ET group, and ET+ chloroquine group. Acute lung injury was induced by intravascular injection of ET (500μg/kg). The arterial gas analyses, leucocyte and platelet counts in peripheral blood, PLA2 activity both in serum and lung tissue, lipid peroxide (LPO) and superoxide dismutase (SOD) in lung tissue were measured. Electron microscope and light microscope were used to observe the pathological injuries in pulmonary tissue. The protective effects of chloroquine in early ALI were evaluated. Results Compared with saline controls, rabbits treated with ET displayed the early lung injuries, such as the decrease of PaO2 (P＜0.05), the decrease of leucocytes and platelets in peripheral blood, the leukocytes sequestration in lung tissue. The PLA2 activity significantly increased in ET group compared with control group and chloroquine group both in serum and pulmonary tissue. In ET group, concentration of LPO increased in lung tissue (P＜0.05), while concentration of SOD decreased (P＜0.05). Severe histopathological injuries were presented in ET group, including pulmonary edema, lung tissue haemorrhage, inflammatory cells infiltration, asphyxial membrand formation, partial pulmonary closure and emphysema.Ultrastructural changes showed both type Ⅰ and type Ⅱ epithelial cells injury in ET group, the edema of endothelial cells, interalveolar septum thickening. In chloroquine group, PaO2 didn't decrease, PLA2 activities in serum and pulmonary tissue were lower than ET group (P＜0.05, P＜0.05), while the concentration of LPO in lung tissue decreased (P＜0.01) and SOD increased significantly (P＜0.01). Pathological examination showed slight pulmonary edema, inflammatory cells infiltration were extenuated, ultrastructural examination proved that the injuries were alleviated by chloroquine compared with ET group. Conclusion Intravascular injection of ET could successfully induce the early ALI models in rabbits. Chloroquine could inhibit the PLA2 activation and reduce the oxidative injury in lung tissue. The experiment result demonstrated PLA2 activation and oxidative stress played important roles in the pathophysiological process of early ET-induced ALI in rabbits.

Objective To study the histopathological features, diagnosis criteria, the relationship of surgery pattern and prognosis of phyllode tumor of breast. Methods To analyze the histopathological features and clinical outcome of different surgery patterns in 203 patients with phyllode tumor of breast by Chi square test, Cluster, Focater, Logistic and Cox multivariate regression according to the request of SPSS 10. 0. Results 203 patients with phyllode tumor of breast were divided into three groups, i.e. benign 133 cases, borderline 42 cases and malignant 28 cases. Local recurrences in three groups were 28, 19 and 18, respectively. The patients died from tumor were 0, 2 and 16, and circulatory metastases were 0, 1 and 10, respectively. Five-year survival were 100 %,92. 0% and 33. 3% in the three groups of 131 patients by a 5 years' follow-up survey. Conclusions Tumor necrosis has important value in the diagnosis of phyllode tumor. Nature of tumor margin, cellular pleomorphism, frequency of mitoses and tumor necrosis were statistically appropriate composition in histological diagnosis of phyllode tumor. Wide local excision is preferred for the benign and borderline phyllode tumor, while simple mastectomy is indicated for recurred borderline and malignant, but tylectomy should be abolished in the treatment of phyllode tumor. Correlation of histotypes of phyllode tumor with local recurrence and tumor death was statistically significant at a level of P＜0. 001; correlation of infiltrative growth of the tumor with local recurrence was statistically significant at a level of P＜0. 001. Tumor necrosis and mitotic activity were independent prognostic factors.

Background Leiomyoma is the most common benign tumor of the esophagus; if malignancy can be ruled out preoperatively, minimally invasive treatment can be performed either thoraco-scopically or laparoscopically. Methods We report on two cases, where we resected a midesophagus tumor by a thoracoscopic and a distal esophageal tumor by a laparoscopic approach under endoscopic assistance during the resection procedure. By this thoracoscopic-endoscopic and laparoscopic-endoscopic rendezvous procedure uncomplicated resection was performed and perforation was avoided. Conclusion Minimally invasive surgical therapy is recommended for the resection of benign submucosal tumors of the esophagus. Tumors located in the middle and distal third can easily be resected. Endoscopic control with rendezvous technique during the operation is recommended to avoid mucosal injury.

Objective In the treatment of acute myeloid leukemia (AML-M2a), the first CCR (continuous complete remission) has been one of the most critical indicators to the prognosis of the patients. Using microarray approaches, gene expression profiles have been studied in patients with different CCR, in order to find out the genes relevant to the progresses of the AML. Methods Bone marrow mononuclear cells were collected and used as different experimental groups respectively. Group A composed of three AML patients with CCR<6 months, while group B composed of three AML patients with CCR>12 months. mRNAs were purified and labeled with Cy3 and Cy5 respectively, which were used to hybridize against the Agilent human 1B 60mer oligonucleotide microarrays. Results In the 20173 genes tested, 21 genes were found expressed differentially between these two groups. Of these differentially expressed genes, 10 genes were up-regulated while 11 genes were down-regulated in group A. Conclusion Through microarray studies, 21 genes including APP were found to be differentially expressed in AML patients whom were treated with standard chemotherapy. Theses genes can be early indicators for the diagnosis as well as prognosis of the refractory AML.

Objective To study the changes in activity of phospholipase A2 (PLA2) in the course of endotoxin (ET) induced acute lung injury (ALI) inrabbits and the antagonizing effect of fructose-1, 6-diphosphate (FDP), in order to evaluate the therapeutic effect of FDP on ET-induced ALI. Methods Flapeared white rabbits were randomly assigned to three groups: control group (group A), ET challenge group (group B) and treatment group (ET challenged followedby FDP, group C). Group A animals were injected with saline (2ml/kg) as control. Group B animals were injected with ET (500μg/kg) solution followed by saline. Total amount of liquid was 2ml/kg. Group C animals were given the same amount of ET solution followed by injection of FDP (300mg/kg) solution. Total amount of liquid was also 2 ml/kg. During the experiment, respiratory rate, heart rate, blood pressure, arterial blood gases and the plasma PLA2 activity were determined at 0h, 0. 5h, 2h, 4h and 6h respectively. The rabbits were sacrificed at 6h, pulmonary PLA2 activity was assessed, and the pathologic changes in pulmonary tissues were examined with light microscope and transmission electron microscope. Results Compared with group A,rabbits of group B manifested the typical characters of ALI after ET injection, and the PLA2 activity in both serum and pulmonary tissue was much higher than those of group A (P＜0. 01).Values of the PLA2 activity in group C were between those of the two former groups. At the rame time, obvious pathological changes indicating lung injury were observed in group B and only mild pathological changes could be discerned in group C. Conclusion Activation of PLA2 activity is an important factor in pathogenesis of ET-induced ALI. FDP can antagonize the PLA2 activity and protect rabbits from early ET induced ALI to a certain extent.

Objectives To explore ventilation induced cytokine production and the role of oxidant stress in lung stretch. Methods Both in vitro and in vivo models of ventilator-induced lung injury (VILI) were used. Alveolar epithelial cells were stretched in vitro to mimic the lung injury in VILI. Rats were ventilated at large tidal volume to produce ventilator-induced lung injury in vivo. A total of 23 inflammatory cytokines were screened with micro gene array in stretched alveolar epithelial cells. Cytokines found to have up-regulated in cells were measured in serum and lung tissue of rats exposed to large tidal volume ventilation. For investigating the intracellular pathway of cytokine up-regulation in VILI, exogenous TNF-α or H2O2 was added to culture media of alveolar epithelial cells. Cytokines were then measured. To explore the role of oxidant stress in VILI, N-acetylcysteine (NAC), as an anti-oxidant, was used in vitro and in vivo. Results We found that transforming growth factor-β1 (TGF-β1 and transforming growth factor-β2 (TGF β2) were up-regulated in stretched alveolar epithelial cells and also in serum of rats with large tidal volume ventilation. Tumor necrosis factor-α (TNF-α) had no effects on TGF-β production in alveolar epithelial cells. Exogenous H2O2, as an oxidant, increased TGF-β production in alveolar epithelial cells. NAC, an anti-oxidant, decreased stretch induced TGF-β production, along with a down-regulation of oxidant injury. NAC also blocked the up-regulation of TGF-β in in vivo model of VILI. Conclusion TGF-β1 and TGF-β2 were up-regulated in VILI. Oxidant injury mediated up-regulation of TGF-β in VILI. NAC, which attenuated oxidant injury and blocked TGF-β up-regulation in VILI, could be a future therapeutic strategy in VILI.

Objectives To evaluate the influence of cardio-pulmonary bypass (CPB) to the endothelial function and lipid oxidative status of the Coronary Artery Bypass Grafting (CABG) patients. Methods Thirty-five coronary patients (female 5 cases, male 30 cases, mean age 59.5±8. 6 years) who needed CABG operation therapy were included. The control group consisted of 20 health volunteers (female 5 cases, male 15 cases, mean age 49. 5 ± 7.3 years).Blood samples were taken at preoperative, 3 and 7 days postoperative. Serum von Willebrand factor (vWF) level, one of the endothelial dysfunction markers,and oxidized low density lipid (OxLDL) level, one of the lipid oxidization status markers were detected with ELISA kits. Results The levels of vWF and OxLDL in CABG patients' serum were elevated significantly compared with those of control group ( 139.60% ± 34. 39% vs 82. 79% ±13. 89% and 63. 7 ±18. 9U/ml vs 25.4± 15.2U/ml respectively). Three and 7 days after CABG significant increases in the vWF level (194. 30% ±37. 74%; 181.07% ±31.05%vs 139.60% ±34.39% at baseline, P＜0. 0001, respectively) and OxLDL level (79. 3±26. 7U/ml; 72.4±23.1U/ml vs 63.7±18. 9U/ml at baseline, P＜0. 01, respectively) were detected. Conclusion Compared with normal people, the endothelial system of CABG patients is impaired, and their lipids are in a more oxidative status. CABG procedure with CPB can further deteriorate the conditions, which may imply that the patients are predisposed to acute coronary accident shortly are the procedure.

Objective Albumin is well known to decrease in response to sepsis, However, the.degradation and distribution in patients with severe sepsis to explore the mechanism of hypoalbuminemia in sepsis. Methods 10 volunteers and 10 patients with severe sepsis. 125I labeled albumin was administered intravenously to 10 healthy volunteers and 10 patients with severe sepsis. Each subject had frequent blood samples taken at 0,1,2,4,8,12,24 hours and on day 2, 3, 4, 5, 6, 7, 9, 11, 13, 15, 18, 22, 25 to measure 125I concentration and draw the curve of concentration over time. Plasma was regarded as the central pool and body fluid as side pool, The curve of albumin concentration vs time was expected to follow two compartment model. Results Radioactivity of blood samples was counted and the results were graphically expressed. The half-life time(t1/2), apparent volume of distribution(Vd) and transportation rate(K12) of albumin from the central pool to the side pool were calculated. The half-life time in sepsis was obviously shorter than that in control group (8.2 1.4 vs 12.5 1.7days, P<0.001). The transportation rate in sepsis group was quicklier than that in control group [(4.4±1.9)× 10-2/h vs (2.4±0.6)×10-2/h, P<0.005]. There was no significant difference in apparent volume of distribution between two groups. Conclusions In patients with severe sepsis, the distribution rate of albumin from plasma to body fluid was obviously elevated and the decomposition rate of albumin was markedly increased.