OBJECTIVEEndothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.

METHODSRAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.

RESULTSIncubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.

CONCLUSIONActivation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.

Animals ; Aorta ; cytology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Arecoline ; Carbachol ; Cell Cycle ; Endoplasmic Reticulum ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Homocysteine ; adverse effects ; Mitochondria ; metabolism ; Rats ; Receptors, Muscarinic ; metabolism

Amphiregulin is the ligand of epidermal growth factor receptor.It's widely expressed in many tissues and is involved in the oncogenesis,progression and metastasis of tumors.In the clinical study of lung caner,amphiregulin is a prognostic marker for NSCLC patients.Furthermore,amphiregulin is closely associated with the sensitivity and resistance of EGFRTKI treatment.Medicine target on amphiregulin can inhibit the activity of tumors.

Objective To observe the effects of recruitment maneuver (RM) and tidal volume with different amount of gas after RM ventilation on lung diastole function in rats with acute lung injury (ALI). Method ALI rat models were induced by intravenous infusion of lipopolysaccharide (LPS) in dose of 6 mg/kg. Twenty-five rats were randomly(random number) divided into control group ( n = 5), ALI group ( n = 5), low tidal volume group (LV group,VT= 6 mL/kg, n = 5), sustained inflation (SI) with low tidal volume (SI+ LV group, VT=6 mL/kg, n = 5), and SI with moderate tidal volume group (SI+ MV group, VT= 12 mL/kg, n = 5). The RM carried out by using SI with airway pressure 30 cmH-2O for 30 seconds, and the positive end-expiratory pressure (PEEP)was set at 5 cmH2O. Lung tissue was taken after mechanical ventilation for 5 hours. The mean arterial pressure (MAP) was monitored throughout the entire course of experiment. Endothelin-1 ( ET-1 ), endothelial nitricoxide synthase (eNOS), and acetylcholine-(Ach-) induced endothelium-dependent relaxation response of isolated pulmonary artery rings were investigated after mechanical ventilation for 5 hours. Results The LPS increased the ET-1 level in lung tissue, decreased the level of eNOS in lung tissue, and impaired the Ach-induced endothelium-dependent relaxation response in pulmonary vassals, without obvious influence on systemic hemodynamics. SI + LV significantly reduced LPS-induced elevation of ET-1 level, and increased the level of eNOS, and significantly lessened endothelial dysfunction and ameliorated dysfunction od endothelium-dependent relaxation in pulmonary vas sals. Conclusions RM with high tidal volume or lowtidal volume ventilation could improve the lung vascular endothelial function of rats with acute lung injury, and RM with low tidal volume ventilation could lessen more the injury of lung vascular endothelial diastole function in rats with acute lung injury.

Objective To analyze the risk factors of liver metastasis from gastric cancer. Methods The clinical data of 225 patients with liver metastasis from gastric cancer who had been admitted to our hospital from January 1996 to December 2001 were retrospectively analyzed. Synchronous liver metastasis was observed in 123 patients and metachronous liver metastasis in 102 patients. The risk factors of liver metastasis from gastric cancer were evaluated. Results The gender of patients, location and size of gastric cancer foci, differentiation and invasion depth of gastric cancer, Lanren classification, lymph node metastasis and lymph node metastasis in region Ⅷ, vascular invasion, extrahepatic metastasis, ascites and peritoneal metastasis were the significant factors associated with liver metastasis from gastric cancer (X2 = 43.560-263. 907, P<0.05). All the factors except the size of gastric cancer foci, extrahepatic metastasis and ascites were found to be the significant factors associated with different types of liver metastasis from gastric cancer (X2 = 6.673-26. 555, P < 0.05 ). Logistic regression analysis demonstrated that the gender of patients, lymph node metastasis and peritoneal metastasis were the factors that determined the types of liver metastasis from gastric cancer. Conclusion The gender of patients, lymph node metastasis and peritoneal metastasis are the important factors to evaluate the occurrence of different types of liver metastasis from gastric cancer.

Objective To explore the occurance and risk factors of recurrence and metastasis of gastric cancer after gastrectomy.Methods From January 2001 to December 2004, the clinic pathological data of 141 patients with recurrence and metastasis after radical gastrectomy in Tianjin Medical University Cancer Institute and Hospital were analyzed retrospectively.The possible clinic pathological factors which may affect tumor recurrence were analyzed.Results After the surgery, the 1, 2, 3 and 5-year cumulative recurrence rates were 58.2%(82/141) 、80.1%(113/141)、89.4%(126/141) 、97.9 % (138/141)respectively.The results of multivariate analysis indicated that the tumor size,invasive depth, lymph node metastasis were independent factors which affected recurrence and metastasis after radical gastrectomy (P = 0.017, 0.003, 0.000).Invasive depth, lymph node metastasis and tumor differentiation degree were independent factors which affected early recurrence and metastasis after radical gastrectomy (P=0.042, 0.000, 0.039).Conclusions The tumor size,invasive depth, lymph node metastasis are the independent risk factors to predict the recurrence and metastasis after radical gastrectomy.Most of the recurrences and metastasis is found within 2 years after radical gastrectomy.Invasive depth, lymph node metastasis and tumor differentiation degree are the independent factors to predict early recurrence after radical gastrectomy.

OBJECTIVETo study the clinical characteristics, diagnosis, strategy of treatment and prognosis of multiple primary cancers in the digestive system.

METHODSAll malignant lesions in the digestive system encountered in the past 10 years were analysed by a computer software.

RESULTS116 cases of multiple cancers were observed with an incidence of 1.74%. Male to female ratio was 2.7:1. There were 62 synchronous carcinomas and 54 metachronous ones. In decreasing sequence, they were: colon > stomach > rectum > liver. The 1-, 3-and 5-year survival rates of the synchronous ones were 42.3%, 23.1% and 11.5%. Those of metachronous ones were 95.2%, 85.7% and 76.2%.

CONCLUSIONHigh frequency of multiple primary cancers is found in the colon, especially over the right side. Metachronous carcinomas usually give better prognosis than the synchronous ones. The longer the interval between the development of the two primary lesions, the better the prognosis. Multiple cancers of the colon have satisfactory prognosis. The key to improve the treatment is to be vigilant against lesions in the digestive tract so as to arrive at a timely diagnosis and treatment. The strategy of combining radical treatment with immuno-supportive therapy is advocated.

Colonic Neoplasms ; pathology ; Digestive System ; pathology ; Female ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; Neoplasms, Multiple Primary ; pathology ; Rectal Neoplasms ; pathology ; Stomach Neoplasms ; pathology ; Survival Analysis ; Time Factors

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Membrane Proteins/*biosynthesis ; Membrane Proteins/genetics ; Membrane Proteins/isolation & purification ; Plasmids/biosynthesis ; Plasmids/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus/chemistry ; SARS Virus/*genetics ; Viral Vaccines/biosynthesis

China ; Disease Outbreaks ; Humans ; Influenza B virus ; Influenza, Human ; epidemiology

OBJECTIVETo study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.

METHODSY1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).

RESULTSY1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.

CONCLUSIONOst could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.

Adrenal Cortex ; drug effects ; Adrenal Cortex Neoplasms ; pathology ; Animals ; Corticosterone ; biosynthesis ; Coumarins ; pharmacology ; Gene Expression ; Mice ; RNA, Messenger ; metabolism ; Tumor Cells, Cultured

Objective To compare the effects of hepatic vein occlusion with tourniquet and Satinsky clamp in reseeting liver tumor involving the second hepatic portal.Methods From Jan 2003 to Jun 2006,180 patients underwent major liver resection with the selective hepatic vascular exclusion (SHVE).According to methods of hepatic vein occlusion,they were divided into two groups:Occlusion with tourniquet(tourniquet group,n=95)and occlusion with Satinsky clamp(Satinsky clamp group,n= 85).In tourniquet group,the hepatic veins were encircled and occluded with tourniquet,and in Satinsky clamp group,the hepatic veins were not encircled and clamped directly with Satinsky clamp.Data regarding the intraoperative and postoperative courses of the patients were analyzed.Results There was no difference between the two groups regarding the operating time,ischemia time,intraoperative blood loss and postoperative complications rate.The dissecting time of hepatic veins was significantly shorter in Satinsky group(6.2?2.4 min vs 18.3?6.2 min).lu the tourniquet group,five hepatic veins(one fight hepatic vein and four common trunk of left-middle hepatic veins)could not be dissected and encircled because of the tumors involving the cava hepatic junction.Another patient's common trunk of left-middle hepatic vein was inadvertently lacerated during the dissection.Hepatic veins in these 6 patients were occluded with Satinsky clamp successfully.Conclusion Occlusion with Satinsky clamping is safer and easier procedure than tourniquets in the resection of liver tumor involving the second porta hepatis.