Objective To explore a simple, effective and stable method for the isolation and purification of Kupffer cells from rat liver, enabling further study on the structure and function of these cells in vitro. Methods After laparotomy, a catheter was inserted into the portal vein and secured with artery clamp. Then, the rat liver was perfused and digested with solution Ⅰ and solution Ⅱ containing 0.05% collagenase Ⅳ respectively. The cell suspension was centrifuged with isopycnic sedimentation in a two-step Percoll gradient to harvest Kupffer cells. The isolated Kupffer cells were purified by selective adherence after 30 min of cultivation, and identified by evaluation of phagocytosis of India ink and peroxidase staining with DAB through light and electron microscopy. Results It was verified that the viability of isolated Kupfffer cells was more than 90% through Trypan blue staining. Those Kupffer cells could attach to plastic quickly and phagocytose ink, and had the appearance of "fried eggs" in positive peroxidase staining with a purity of 95%. Under the light microscopy, the appearance of newly isolated Kupffer cells was round with uniform shape and size. After two days of culture, Kupffer cells appeared to distend with irregular or stellate shape. The typical features were observed in the transmission electron micrographs. There were numerous pseudopods and occasional cup-like indentations in the cell membrane of Kupffer cells. The cytoplasm contained numerous types of lysosomes and other phagocytotic vesicles. Conclusion The method for isolating and culturing Kupffer cells in this study is effective and stable, and the biological characters are preserved in the cultured cells.

Objective To study the effect of preoperative selective arterial infusion chemotherapy (PSAIC) on large intestine cancer. Methods 63 patients with colorectal cancer underwent PSAIC with 5 fluorouracil,mitomycin and E adriamycin; the changes of clinical manifestation and pathology were observed and analyzed. Results (1)clinical manifestation:abdominalgia relieved in 18 patients and abdominal distention relieved in 16 patients.Improvement of hematochezia was found in 7 patients.In addition, of 13 patients with partial ileus, the clinical symptoms relieved in 9 patients. (2) pathology: there were karyopyknosis,karyorrhexis,coagulation and necrosis of cytoplasm in cancer cells. infiltration of inflammatory cells, edema and fibroelastosis in mesenchyne of cancer tissue. Proliferous intima and thrombus were also observed in the vessels. Most of these changes were moderate,and marked changes could also be seen in necrosis of cytoplasm and vessel intima proliferation. Conclusion PSAIC can control the focus of primary disease and the micrometastatic foci as well as improve the clinical symptoms, such as intestinal obstruction, and hematochezia, so PASIC is helpful to subsequent operation,and can improve the survival rate of patients with colorectal cancer.

AIM:To investigate the effect of fluvastatin on the expression of serum and glucocorticoid inducible kinase 1 ( SGK1) and connective tissue growth factor ( CTGF) induced by aldosterone ( Ald) in rat mesangial cells (GMCs). METHODS:GMCs were divided into (1) control group; (2) aldosterone group with different concentrations and times; (3) Ald (10 -7 mol/L) + spironolactone (10 -9 mol/L) group; (4) Ald (10 -7 mol/L) + LY294002 (20 ?mol/L) group; (5) Ald (10-7mol/L) +SB203580 (20 mmol/L) group; (6) the group of Ald (10-7mol/L) + fluvas-tatin at different concentrations (10-7,10-6,10-5 mol/L); (7) Ald (10 -7mol/L) + fluvastatin (10 -5mol/L) + mevalonate (10 -4 mol/L) group; (8) Ald (10 -7 mol/L) + fluvastatin (10 -5 mol/L) + FPP (farnesyl pyrophosphate,10-4 mol/L) group; (9) Ald (10 -7mol/L) + fluvastatin (10 -5 mol/L) + GGPP (geranylgerany pyrophosphate,10 -4 mol/L) group. The protein levels of SGK1 and CTGF were determined by Western blotting. The levels of fibronection (FN),monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) in the supernatants were determined by enzymelinked immunosorbant assay (ELISA). RESULTS:Aldosterone stimulated the protein expression of SGK1 and CTGF in cultured mesangial cells in a dose-dependent manner (P

Humans ; Infant ; Male ; Tuberous Sclerosis ; Twins

Animals ; Bufo bufo ; Caspase 3 ; genetics ; metabolism ; Female ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Objective To explore the clinical effects of unilateral placement of plastic stents by ERCP for nonresectable Bismuth type IV hilar cholangiocarcinoma.Methods A prospective study was conducted in 42 patients with nonresectable Bismuth type IV hilar cholangiocarcinoma,who had unilateral insertion of plastic stents in the recent 4 years,All of the patients had successful insertion of single plastic stent by ERCP manipulation.Early results (less than 30 days) of procedure-related complications,mortality,long-term results (greater than 30 days) and survival were observed.Results All of the 42 patients had successful drainage.Mean total bilirubin value decreased from (332.3?163.4)?mol/L to (30.6?18.5)?mol/L,and complete resolution of jaundice was achieved in 61.9% (26/42)of the patients.Early comlications included papilla bleeding due to EST in 4 of 42 patients (9.5%) and acute cholangitis in 10 of 42 patients (23.8%).No procedure-related death occurred.All of the patients needed to have their stents replaced periodically,and the median duration up to the first stent replacement was 65 d,and without significant differences between the two groups in stent insertion to the left and right hepatic duct.Median duration of stent patency was 5.15 months,and median patient survival was 6.5 months,but also no significant differences were found between the two groups.Conclusions Unilateral plastic stent insertion is safe and feasible,can achieve adequate drainage for relatively long time,and can improve life quality and survival for patients with nonresectable hilar cholangiocarcinoma.

Objective To investigate the diagnostic value of coding gene of Sj26, Sj32 and Sj14-3-3 amplified by PCR for chronic Schistosomiasis japonica. Methods The DNA was extracted from sera of 40 patients with chronic Schistosomiasis japonica, the coding gene of Sj26, Sj32 and Sj14-3-3 was amplified by PCR and identified by 1.2% agarose gel electrophoresis. DNA from the sera of 21 patients with Clonorchiasis sinensis, 13 patients with Parogonimiasis westermani and 43 healthy donors was taken as control. Results A total of 399 bp coding gene of Sj14-3-3 was amplified successfully from sera of the patients with chronic Schistosomiasis japonica,but Sj26(676 bp) and Sj32( 1270 bp) coding gene were not obtained. Control groups were all negative. Conclusions Sj14-3-3 coding gene amplified by PCR can be used for genetic diagnosis of chronic schistosomiasis.

Objective To study the diagnostic value of rSj26-Sj32-IgG-ELISA for acute schistosomiasis japonica. Methods Purified rSj26-Sj32 fusion protein and crude Schistosoma japonicum antigen (SjAWA)were used to establish IgG-ELISA to detect serum of patients with acute schistosomiasis, and clonorchiasis sinensis,paragonimiasis westermani, alveolar echinococcosis, cystic echinococcosis, type B hepatitis, lung tuberculosis patients and healthy human serum were used as control. Results The sensitivity and specialty were 90.00%(45/50) ,97.67% (42/43) and 92.00% (46/50),97.67% (42/43) in detection of acute schistosomiasis japonica with rSj26-Sj32and SjAWA, respectively, and the difference was not statistically significant(x2 were both 0.0, all P ＞0.05). The serum cross-reaction reactivity was 20.00%(2/10) in patients with alveolar echinococcosis with SjAWA,but no cross-reaction with rSj26-Sj32, the difference was not statistically significant(x2 = 0.5, P ＞ 0.05). The serum cross-reactivity were 14.29% (3/21 ), 7.69% (1/13) and 19.05% (4/21 ), 7.69% (1/13) among patients with clonorchiasis sinensis and paragonimiasis westermani by rSj26-Sj32 and SjAWA, but no cross reaction with type B hepatitis and lung tuberculosis patients, the difference was not statistically significant (x2 were both 0.0, all P ＞ 0.05). The positive predictive value, the negative predictive value and the diagnostic efficiency with acute schistosomiasis japonicum by rSj26-Sj32-IgG-ELISA and SjAWA-IgG-ELISA were 97.83% (45/46),89.36% (42/47),93.55% (87/93)and 97.87% (46/47),91.30% (42/46),94.62% (88/93), respectively, and the difference was not statistically significant (x2 were both 0.0, all P ＞ 0.05). Conclusion rSj26-Sj32 fusion protein can be used for the immune diagnosis of acute schistosomiasis japonica.

Aim To study the protective effect of tetra-hydroxystilbeneglucoside ( TSG ) on cardiac injury and the mechanism involved in silent mating type informa-tion regulation 2 homolog 1 ( SIRT1 ) and adenosine monophosphate-activated protein kinase( AMPK) in the diabetic rats. Methods Type 2 diabetic rats were sac-rificed after administration with TSG for 8 weeks. Blood glucose, blood lipids, liverfunction,creatine ki-nase ( CK ) , lactate dehydrogenase ( LDH ) as well as myocardial nonesterified fatty acids( NEFA) were deter-mined by using biochemical test. The concentration of myocardial fatty acid transport proteins ( FATPs ) and-fatty acid β-oxidase ( FA-β-oxidase ) , and the levels of tumor necrosis factor alpha ( TNF-α) , interleukin -6 ( IL-6 ) , interleukin-1β( IL-1β) in serum were also measured by ELISA method and radio immunoassay re-spectively. The protein expressions of TNF-α, IL-6, IL-1β, SIRT1 and AMPK were detected by Western blot. Results Treatment of TSG reduced the contentof blood lipids, NEFA and collagen without affecting the content of blood glucose and insulin. The levels of TNF-α, IL-6 and IL-1βin serum as well as the protein expressions of TNF-α, IL-6 and IL-1β of cardia were also inhibited by administration with TSG. Treatment of TSG caused a significantly increased concentration of myocaidial FATPs and FA-β-oxidase, and dramatically restored the decreased protein expressions of SIRT1 and pAMPK in diabetic rats. Conclusion The protec-tive mechanisms of TSG against diabetic rats are in-volved in the alleviation of inflammatory mediator injury and improving energy metabolism.