AIM To study the effect of genistein (GST), a protein tyrosine kinases inhibitor, on L-type calcium channel currents (Iba,L or Ica,L, dependent on permeating ion used) in freshly dispersed colon smooth muscle cells from guinea-pig. METHODS Single colon smooth muscle cells were enzymatically dissociated from guinea-pig. L-type calcium currents were measured by conventional whole-cell patch-clamp techniques. RESULTS The peak amplitudes of Iba,L elicited to 10 mV test potential from a holding potential of -80 mV, were reversibly and dose-dependently reduced by GST (10-100 μmol·L-1) with an IC50 value of (39.9±3.6)μmol·L-1. Bath application of GST shifted the steady-state inactivation curves of Iba,L in a hyperpolarized direction (about 10 mV, P＜0.01) without altering their slopes. The peak amplitudes of Iba,L were also inhibited but to a less extent by daidzein, an inactive analogue of GST. Sodium orthovanadate 1 mmol·L-1, a potent inhibitor of protein tyrosine phosphatases, blocked GST-induced inhibition of Ica,L. CONCLUSION GST can block L-type calcium channel activity in guinea-pig colon smooth muscle cells via tyrosine kinase pathway.

Objective To observe the expressions of autophagy-related proteins Beclin-1and apoptosis-related pro-teins Bcl-2 in CA1 area of hippocampus in vascular dementia model rats. Methods The experimental rats were randomly divided into sham operation group and vascular dementia model group. The model group was divided into five subgroups:1, 2, 4, 8 and 12-w group (n=6 for each group). The vascular dementia rat model was established by blocking four vessels. Learning and memory abilities were detected by Morris water maze. The expressions of Beclin-1 and Bcl-2 were detected by immunohistochemistry assay. Results The positive expressions of Beclin-1 and Bcl-2 were detected at 1-w in CA1 area of hippocampus in model rats, which reached the peak at 4-w and began to decline in 8-w, maintained the higher levels at 12-w. There were significantly higher expressions of Beclin-1 and Bcl-2 at all-time points in model group than those of sham operational group (P＜0.05 or P＜0.01). The positive expression of Beclin-1 was positively correlated with Bcl-2 at all-time points in CA1 area of hippocampus in model group (r=0.809, P＜0.05). Conclusion Results showed that autophagy and apoptosis occurred simultaneously and expressed the same trend in CA1 area of hippocampus of vascular dementia mod-el rats.

AIM: Stable human macrophage-derived foam cell model is significant for the study on artherosclerosis. This study investigated the feasibility of establishing macrophage-derived foam cell model using U937 cell lines. METHODS: The experiment was performed at Institute of Basic Medicine, Chengde Medical College from March to September 2006. ①U937 cell lines were purchased from Institute of Biochemistry & Cell Biology, Chinese Academy of Sciences. ②Sixteen bottles of U937 cells (109 L-1) were incubated at 37 ℃ in saturated humidity containing 5% CO2 for 72 hours. Among them, eight bottles contained 100 ?g/L phorbol-12-myristate-13-acetate (PMA) and 100 mg/L low-density lipoprotein (LDL) as experimental group, and the other eight bottles only 100 mg/L LDL as control group. ③Cell morphology was studied under light microscope by Wright's and Oil red O staining. Cell total cholesterol (TC) was measured after 72 hours of incubation. RESULTS: A large amount of lipid droplets were found in the cytoplasm by Oil red O staining in cells of the experimental group, but not found in control group cells. TC in cells of the experimental group was significantly higher than in control group [(520.13?37.52), (39.47?9.26) mg/g, t=35.18, P

Objective To evaluate the impact of preoperative renal artery embolization in renal cell carcinoma patients with venous tumor thrombus.Methods A total of 249 RCC patients with venous tumor thrombus underwent radical nephrectomy and thrombectormy in our hospital.Seventy-four patients received preoperative renal artery embolization while other 175 patients did not.The related items were analyzed.The tumor thrombus level was divided into 5 groups with American Mayo clinic classification system.Considering the significant difference in tumor situation and treatment strategies, we took the hepatic vein as a cut-off line, dividing patients into two subgroups, the early and advanced tumor thrombus groups.There were 208 patients in the early tumor thrombus group and 41 patients in the advanced group.The related items were analyzed respectively.Results Patients in the embolization group tended to have larger tumors and higher percentage of advanced tumor thrombus.For all patients, the embolization group had longer operation time [(4.8 ±2.1) h vs.(4.1 ±2.2) h ,P <0.05] and greater median blood loss (900ml vs 500 ml,P <0.05), compared to the non-embolization patients, and RAE did not show benefits on perioperative outcomes seemingly.However, subgroup analysis of patients with advanced tumor thrombus ( above hepatic vein) , in spite of the larger size of tumor, preoperative RAE showed significant benefits on reducing operation time [(7.0 ±1.7)h vs.(8.2 ±1.8)h, P<0.05)] and intraoperative blood loss (2 000 ml vs.3 000 ml, P<0.05) and decreasing blood transfusion (1 525 ml vs.2 050 ml ,P>0.05).Conclusion Preoperative RAE may be more appropriate for patients with advanced tumor thrombus because of its benefits in reducing operation time, intraoperative blood loss and elevating the operative security.

Objective To analyze the clinical manifestations and treatment for toxic megacolon induced by drastic cathartics inpatients with an unknown history of ulcerative colitis. Methods The clinical data of 5 patients with toxic megacolon induced by ulcerative colitis with initial onset type from June 2003 to October 2008 were analyzed retrospectively. Results In 5 cases, the first symptom was abdominal pain and distention. After taking cathartics, these 5 cases were complicated with toxic megaeolon and 2 cases suffering from intestinal perforation. Four female patients suffered from transient unconsciousness, in which 3 patients were found with cerebral lacunal infarction identified by magnetic field diffusion-weighted imaging. All 5 cases underwent exploration, colectomy and ostomy, one patient died perioperatively, anastomotic fistula and anastomotic constriction developed in one each cases. Conclusions The most common clinical manifestations of toxic megacolon induced by ulcerative colitis are abdominaigia, abdominal distention. Emergency therapeutic strategy consists of partial culectomy and ostomy.

Objective To compare the efficacies of the treatment of sternal fracture with wire fixation and the titanium sternal fixation system. Methods Thirty patients with sternal fracture from May 2003 to July 2009 were followed up. Among them,there were 20 patients with wire fixation (wire fixation group), 10 patients with the titanium sternal fixation system (titanium sternal fixation system group). The conditions before, during and after operation,complications and effects were compared to evaluate the effieaeies of titanium sternal fixation system. Results The operative time of titanium sternal fixation system group and wire fixation group were (67.0 ± 7.9) min and (90.0 ± 8.6) min, the blood loss were (11.0 ± 5.4) ml and (48.0 ± 8.4)ml,the duration of drainage were (0.5 ± 0.4) days and (1.9 ± 0.7) days,the amount of drainage were (1.9 ± 1.3) ml and (19.0 ± 4.6) ml, the average hospitalized days were (2.3 ± 0.5) days and (6.9 ± 0.9) days, the duration of pain were (1.5 ± 0.5) days and (3.8 ± 1.1) days, there were all significant difference between two groups (P < 0.05). The rates of wound infection, delayed union or nonunion, re-fracture,plate fracture or plate shift of wire fixation group were 5% (1/20) ,5% (1/20) ,5% (1/20), 10% (2/20). But the rates of titanium sternal fixation system group were 0, there were all significant difference between two groups (P < 0.05). Conclusion The treatment of sternal fracture with titanium sternal fixation system is a simple and stable fixation,high bone union rate and few complications,especially for the sternal fracture.

Objective To clone and express the Tp0453 antigen immuno-dominant epitope fragment of Treponema pallidum (Tp) in Escherichia coli,in an effort to develop serological tests with increased specificity for the diagnosis of syphilis.Methods The gene encoding Tp0453 recombinant outer membrane protein fragment was amplified by polymerase chain reaction (PCR),and inserted into expression vector pQE30 after T-A cloning,then confirmed by restriction map.The constructed recombinant plasmid pQE30-Tp0453 was transformed to E.coli M15 for expression induced by isopropyl β-D-1-thiogalactopyranoside.The expressed product was identified by Western blot,and purified by Ni2+-NTA agarose column chromatography.A double antigen sandwich enzymelinked immunosorbent assays (ELISA) was established by using the recombinant Tp0453 protein to test sera from 48 patients with positive Treponema pallidum particle agglutination test (TPPA),and 40 negative sera as control.Results The PCR amplicon of the target gene was about 490 bp.The recombinant plasmid pQE30-Tp0453 was correctly constructed and successfully expressed in E.coli M15.The expressed product,with a relative molecular of about 21 000,existed in a form of inclusion body,accounting for about 18％ of total somatic protein,and reached a purity of more than 95％ after purification.Western blot showed specific reaction of the expressed protein with Tp positive serum.The ELISA tests with the 88 clinical samples yielded a sensitivity of 97.9％ (47/48),and specificity of 100.0 ％ (40/40).The consistency of results between the ELISA test and the TPPA test was 98.9 ％ (87/88).Conclusion The expressed Tp0453 fragment has showed good immunoreactivity with serum from patients with syphilis,providing the foundation of further development of serological diagnostic kit with increased specificity for the diagnosis of TP infection.

Objectives To study the correlations between stroke-preventive knowledge,health belief and health behaviors in community hypertensive patients.Methods The questionnaire of SPKQ,CHBMS and HPLPⅡwere used to take the investigation among 94 hypertensive patients from a community hospital in Guangzhou.Results The total score on SPKQ was 62.70±18.39 and the average scores on CHBMS and HPLPⅡwere 3.51±0.24 and 2.48±0.37,respectively.The stroke-preventive knowledge was positively correlated with health belief,health motivation and self-efficacy(r=0.289,P<0.01;r=0.246,P<0.05;r=0.350 (P<0.01,respectively).The health motivation was positively correlated with health behaviors(r=0.304,P<0.01)and the seriousness negatively correlated with health behaviors(r=-0.279,P<0.01).Conclusion Medical staff should provide much more stroke education with community hypertensive patients and promote patients’health motivation and self-efficacy of health belief in stroke prevention,help patients understand stroke seriousness,establish and sustain healthy lifestyles.

Objective To evaluate the effects of aspirin-triggered lipoxin A4 (ATL) on lipopolysaccharide (LPS)-induced acute lung injury in mice.Methods Thirty male SPF BALB/C mice,aged 10-12 weeks,weighing 25-30 g,were randomly assigned into 3 groups (n =10 each):normal saline group (group NS),LPS group and ATL groups.ATL 0.1 ml was injected via the tail vein 1 h after intra-tracheal instillation of 3 mg/kg LPS in LPS group.In ATL group,ATL 0.2 mg/kg was injected via the tail vein 1 h after intra-tracheal instillation of 3 mg/kg LPS.At 24 h after instillation,the mice were sacrificed.Bronchoalveolar lavage fluid was collected for determination of the total cell count,proportion of the polymorphonuclear leukocytes,proportion of the mononuclear leukocytes,and concentrations of the total protein,TNF-αt,IL-6,monocyte chemoattractant protein-1 (MCP-1) and IL-10.Lungs were removed for determination of myeloperoxidase (MPO) activity,phosphorylation of p38 mitogenactivated protein kinase (p38 MAPK),c-Jun N-terminal kinase (JNK),and extracellular signal-regulated kinase 1/2 (ERK1/2) in lung tissues and for microscopic examination.The pathological changes of lungs were scored.Results Compared with NS group,the lung injury scores,total cell counts,proportion of polymorphonuclear leukocytes,and concentrations of TNF-α,IL-6 and MCP-1 were significantly increased,and the proportion of mononuclear leukocytes was decreased in LPS and ATL groups,and IL-10 concentrations were decreased,and the concentrations of the total protein,MPO activity,and phosphorylation of p38 MAPK,JNK and ERK1/2 were significantly increased in group LPS (P ＜ 0.05),and no significant change in the concentrations of the total protein,MPO activity,phosphorylation of p38 MAPK,JNK and ERK1/2 was found in group ATL (P ＞ 0.05).Compared with LPS group,the lung injury scores,total cell counts,proportion of polymorphonuclear leukocytes and the concentrations of the total protein,TNF-α,IL-6 and MCP-1 were significandy decreased,the proportion of mononuclear leukocytes and IL-10 concentration were increased,and MPO activity and phosphorylation of p38 MAPK and JNK were decreased (P ＜ 0.05),and no significant change in the phosphorylation of ERK1/2 was found in group ATL (P ＞ 0.05).Conclusion ATL can ameliorate LPS-induced acute lung injury by inhibiting activations of p38MAPK and JNK signal pathways in mice.

Objective:To disiuss the application of liver free technique in renal cell carcinoma patients with Mayo Ⅱ-Ⅳ tumor thrombus.Methods:The clinical data of renal cell carcinoma patients with MayoⅡ-Ⅳ IVC tumor thrombus in our hospital from January 2014 to December 2019 were retrospectively analyzed. 25 patients underwent right part of liver or hepatic portal part dissection via open abdominal approach. There were 20 males and 5 females, aged 45-74 years (mean 61±6 years). All patients underwent urinary tract CTU or MRU examination, vena cava enhanced magnetic resonance angiography.There were left 8 cases, right 17 cases; the median length of tumor was 7 cm (3.6-12.1 cm). There were 1 case of Mayo grade Ⅱ tumor thrombus, 7 cases of Mayo grade Ⅲ tumor thrombus, and 17 cases of Mayo grade Ⅳ tumor thrombus. There were 7 cases of distant metastasis, including 6 cases of lung metastasis and 1 case of bone metastasis. After multi-disciplinary consultation (MDT), 19 patients underwent radical nephrectomy and 6 patients underwent tumor reducing nephrectomy. During the operation, the ligaments around the liver were completely dissociated and the space between the liver and kidney was opened. The bare area of the liver was fully dissociated, to expose the inferior vena cava. For Mayo grade Ⅳ tumor thrombus, 11 cases were treated with free diaphragmatic thrombus removal without thoracotomy, and 6 cases were treated with open chest cardiopulmonary bypass.Results:The median operation time was 444(258-694)min, the median intraoperative blood loss was 2 000(250-10 000)ml, and the median value of suspended red blood cell transfusion was 1 300(400-10 400)ml. The median postoperative hospital stay was 10(4-25)days.15 patients (60%) had postoperative complications, including 8 cases of liver injury, 5 cases of respiratory complications, 4 cases of kidney injury, 3 cases of anemia, 3 cases of infection and 1 case of thrombosis. Three patients died during perioperative period.Conclusions:The application of total liver free technique might obtain good exposure of surgical field, effectively control the hemorrhage of inferior vena cava, which is helpful for safe resection of tumor.