OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

Objective To investigate the treatment and mechanism of aminoguanidine in retina of diabetic of rats .Methods To-tal 60 rats were divided into control group(n=20) ,diabetic group(n=20) and aminoguanidine treatment group(n=20)which would be treated by aminoguanidine for 14 days .Then the eye tissue of rats were took after 14 days administration for pathological obser-vation(HE staining) ,and the induced nitric oxide synthase(iNOS) ,endothelial nitric oxide synthase(eNOS) ,nerve type of nitric ox-ide synthase(nNOS) level and the expression of differences content and expression were investigated by ELISA ,Western blot and PT-PCR .Results HE staining showed that retinal tissue defects decreased and neuronal cells of rats in aminoguanidine treatment group were increased and significant (P<0 .05) compared rats in diabetic group .The iNOS content and expression of rats in amin-oguanidine treatment group were lower than diabetic group by ELISA ,Western blot and PT-PCR ,it was significantly difference (P<0 .05) and without significant difference between the normal group and diabetic group (P> 0 .05) .Compared with diabetes group ,iNOS ,eNOS ,nNOS protein expression in the rat retina in aminoguanidine treatment group were reduced (P< 0 .05) ,and without significant difference between the normal group and aminoguanidine treatment group (P>0 .05) .The iNOS mRNA expres-sion was lower than that of eNOS mRNA and nNOS mRNA in aminoguanidine treatment group .Conclusion Aminoguanidine can improve retinal tissue of diabetic rats with lesions ,the pathways may be selectively inhibit inducible nitric oxide synthase activity of iNOS .

OBJECTIVETo study the clinical pathologic and immunohistochemical features of gastrointestinal mesenchymal tumors (GIMTs), and to investigate the value of molecular markers in GIMTs clinical differentiation diagnosis.

METHODSThe clinical and pathological data of 210 cases of GIMTs, collected from Jan. 1987 to Dec. 2005 in our hospital, were investigated retrospectively. GIMTs were rediagnosed by using standard immunostaining technique in paraffin-embedded tissue. The expression level of CD117, CD34, Desmin, SMA and PS100 were detected by immunohistochemical method.

RESULTSAmong 210 cases of GIMTs, 127 cases were Gastrointestinal stromal tumors (GISTs) (60.5%), 33 leiomyomas and leiomyosarcomas (15.7%), 27 neurogenic tumours (12.8%), and 23 miscellaneous tumors (11.0%). The incidences of GIST, leiomyoma and leiomyosarcoma were similar among men and women. Men were more likely to develop neurogenic tumors and miscellaneous tumors than women. Of all the GISTs, 51.2% cases originated from stomach, 19.7% from small intestine, 11.0% from esophagus, 10.2% from colon and rectum. The most common location of leiomyomas and leiomyosarcomas was esophagus (45.5%). The most common location of neurogenic tumors was retroperitoneum (74.1%). Common symptoms of GISTs included digestive tract hemorrhage in 36 cases (28.3%), abdominal pain in 27 cases (21.3%) and abdominal mass in 24 cases (18.9%). Other GIMT cases except GISTs had no first symptom of digestive tract hemorrhage. It was noticed that 79.5% of GISTs had no obvious invasion, and 72.7% of leiomyomas and leiomyosarcomas had no obvious invasion. 33.3% of neurogenic tumors invaded the adjacent organs or tissues. No metastases had been found in other GIMT cases except GISTs. The neoplastic cells of GISTs were composed of various percentage of spindle (72.5%), epithelioid (11.8%) and mixed-type cells (15.7%). The percentage of spindle cells in leiomyomas and leiomyosarcomas was 94. The immunohistochemical results of GISTs showed that the positive rate of CD117 was 93.7%, CD34 was 69.3%, Desmin was 13.4%, SMA was 12.6%, and PS100 was 10.2%. The immunohistochemical results of leiomyomas and leiomyosarcomas showed that the positive rate of Desmin was 78.5%, SMA was 63.6%, while as the expressions of CD117, CD34, and PS100 were negative. Diffuse strong positive staining of PS100 was observed in 88.9% of neurogenic tumor patients.

CONCLUSIONSGISTs are the most common tumors among GIMTs. GISTs are different from neurogenic tumors, leiomyomas and leiomyosarcomas in initial symptom, tumor location, biological behavior and immunophenotype. Immunohistochemistry plays an important role in differentiating GISTs from leiomyomas and neurogenic tumors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Child ; Child, Preschool ; Female ; Gastrointestinal Neoplasms ; pathology ; Humans ; Immunohistochemistry ; Infant ; Male ; Mesenchymoma ; pathology ; Middle Aged ; Young Adult

AIMTo compare the effects of the non-mitogenetic human acidic fibroblast growth factor (nmhaFGF) and the human acidic fibroblast growth factor (haFGF) on the proliferation and MAPK signal transduction pathway of the malignant tumor cell and to study the clinical safety of nmhaFGF.

METHODSThe mammary tumor cells (MCF-7) were treated with haFGF and nmhaFGF separately. The mitogenic activities of both haFGF and nmhaFGF were detected by MTT method and the cell cycle was analyzed by flow cytometer (FCM). The expression levels of the signal proteins, Grb2 (growth factor receptor bound 2) and ERK1/2 (extracellular signal-regulated kinase 1/2), were detected by semi-quantitative Western blotting method.

RESULTSThe mitogenic activity of nmhaFGF was obviously lower than that of haFGF. The activity of nmhaFGF was weaker than that of the haFGF. The ratio of G1/G0, G2/M of haFGF was markedly lower than that of nmhaFGF and control group, and was reverse in S phase. The expression levels of both Grb2 and ERK1/2 of the nmhaFGF treated group were lower than that of the haFGF treated group and approaching the control group.

CONCLUSIONThe mitogenic activity of the nmhaFGF decreased remarkably. Its mechanism probably via down-regulation of the expression of the signal moleculars, MAPK-ERK1/2 and Grb2.

Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Down-Regulation ; Female ; Fibroblast Growth Factor 1 ; genetics ; pharmacology ; GRB2 Adaptor Protein ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitosis ; drug effects ; Mutation

Osteonecrosis of the femoral head is frequently observed in patients treated with excessive corticosteroids. However, the pathogenesis of corticosteroid-induced osteonecrosis remains unclear. The purpose of this study was to investigate the role of Toll-like receptor 4 (TLR4) signaling pathway in steroid-induced femoral head osteonecrosis in rats. Male Sprague-Dawley rats were injected intramuscularly with 20 mg/kg methylprednisolone (MP) for 8 weeks, twice per week. The animals were sacrificed at 2, 4 and 8 weeks after the last MP injection, respectively, and then allocated to the 2-, 4- and 8-week model groups (n=24 each). Rats in the control group (n=12) were not given any treatment. Histopathological analysis was performed and the concentration of tartrate-resistant acid phosphatase (TRAP) in plasma was determined. The activation of osteoclasts in the femoral head was assessed by TRAP staining. The expression of TLR4, MyD88, TRAF6 and NF-κB p65 that are involved in TLR4 signaling, and MCP-1 production were detected by using real-time PCR (RT-PCR) and Western blotting. The results showed that the osteonecrosis in the femoral head was clearly observed and the concentration of TRAP in the plasma was increased in the model rats. The femoral head tissues in MP-treated rats were positive for TRAP and the intensity of TRAP staining was greater in MP-treated rats than in control rats. As compared with the control group, the mRNA expression of TLR4 signaling-related factors was enhanced significantly at 4 and 8 weeks, and the protein levels of these factors increased significantly with time. It was concluded that MP could induce the femoral head osteonecrosis in rats, which was associated with osteoclast activation via the TLR4 signaling pathway. These findings suggest that TLR4 signaling pathway plays a pivotal role in the pathogenesis of steroid-induced osteonecrosis.

Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

Objective To observe the influences of uncoupling protein 2 (UCP-2) rs660339 variants transfection on cell proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC). Methods Two UCP-2 green fluorescent protein (GFP) lentivirus constructs were created with the rs660339 locus carried C or T (UCP-2C or UCP-2T), respectively. HUVEC were cultured after lentiviral infection of UCP-2C or UCP-2T. The expression of UCP-2C or UCP-2T was detected with real time polymerase chain reaction. Cell proliferation and cell apoptosis were compared among negative control (NC) group, UCP-2T group and UCP-2C group using CCK-8 cell viability and flow cytometry. Western blot and immunostaining were employed to examine the expression of Bcl-2 gene. Results The lentivirus constructs were successfully created.>80%of the transfected cells were found to express GFP under fluorescent microscope. The mRNA levels of UCP-2 gene were significantly increased (F=29.183, P=0.001) in the UCP-2T group and UCP-2C group. The CCK-8 assay revealed that on day two (F=15.970, P=0.004), day three (F=16.738, P=0.004), day four (F=5.414, P=0.045) post-infection, UCP-2T and UCP-2C group showed significantly greater proliferation than the NC cells. The apoptotic rate in the UCP-2T and UCP-2C group was significantly lower than NC group (F=277.138, P=0.000), and the apoptotic rate of UCP-2T was significantly lower than that of UCP-2C (P=0.003). The protein levels of Bcl-2 in the UCP-2T and UCP-2C group were significantly greater than that in the NC group (F=425.679, P=0.000), and the Bcl-2 expression of UCP-2T was greater than that of UCP-2C (P=0.002). The Bcl-2 density in the UCP-2T and UCP-2C group were greater than that in the NC group (F=11.827, P=0.008), while there was no difference between UCP-2T and UCP-2C group (P=0.404). Conclusion The variants of UCP-2 rs660339 may influence HUVEC proliferation and apoptosis, and UCP-2T showed a stronger effect of inhibiting apoptosis than UCP-2C.

OBJECTIVETo evaluate the effect of Sapylin combined with intraperitoneal and systemic chemotherapy for advanced colon cancer following radical operation on local recurrence, hepatic metastasis, and overall survival rate.

METHODSFrom Jan. 2004 to Dec. 2005,132 patients with stage II or III colon carcinoma after radical operation were randomly divided into two groups: Sapylin combined with chemotherapy(Sapylin) group and the control group. Toxic reaction, local recurrence, hepatic metastasis, and overall survival rate between two groups were compared.

RESULTSBoth groups successfully completed the trial. There was no significant difference in toxic reaction between two groups, the recurrence and hepatic metastasis rate in Sapylin group were lower than those in the control group(9/60, 15.0% vs. 22/72, 30.6%; 11/60,18.3% vs. 22/72, 34.7%, respectively), which were statistically significant (P<0.05 respectively). The 3- year survival rate in Sapylin group was higher than that in control group(73.3% vs. 54.2%), which was statistically significant (P<0.05).

CONCLUSIONSapylin combined with intraperitoneal and systemic chemotherapy can effectively decrease local recurrence and hepatic metastasis,and improve the survival in patients with advanced colon cancer following radical operation.

Adult ; Aged ; Biological Products ; therapeutic use ; Chemotherapy, Adjuvant ; Colonic Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Postoperative Period ; Prospective Studies ; Streptococcus pyogenes ; Survival Rate

OBJECTIVETo study the short-term results of metal on metal hip resurfacing arthroplasty (HRA) for the treatment of patients with ankylosing spondylitis (AS), to explore the indications and technology of this surgery.

METHODSFrom February 2006 to April 2008, 11 patients (15 hips) with ankylosing spondylitis were treated with metal on metal total hip resurfacing arthroplasty. Among them, 9 patients were male, and 2 patients were female, with an average age of 32.5 years (ranged from 16 to 53 years). The comparative study on preoperative and postoperative pain, range of motion, correction of deformity and function evaluation were performed. The preoperative and postoperative Harris scores were compared and the feasibility and technical difficulty were analyzed.

RESULTSTen patients were followed up for an average period of 16.2 months (ranged from 8 to 34 months) and 1 patient was lost. Pain disappeared in 10 patients. There were no heterotopic ossification, no femoral neck fracture, no dislocation, no infection and no revision in all patients. From preoperation to present, the mean flexion angle of hip was improved from preoperative 0 degrees to 75 degrees to postoperative 35 degrees to 105 degrees; the mean abducting angle of hip was improved from preoperative 0 degrees to 30 degrees to postoperative 15 degrees to 55 degrees; and the average Harris hip score improved significantly from (30.9 +/- 3.4) (2-47) to (85.1 +/- 3.1) (46-94). According to evaluation criteria, 10 hips got an excellent result, 3 good and 1 poor.

CONCLUSIONThe total hip resurfacing arthroplasty is an effective solution for the treatment of the younger and active patients with AS and it shows satisfactory short-term results. It is very significant to analysis the different conditions of patients so as to choose proper strategy.

Adolescent ; Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Range of Motion, Articular ; Spondylitis, Ankylosing ; physiopathology ; surgery ; Young Adult

OBJECTIVETo investigate the antioxidant effect of melatonin (MT) in the rats with phosgene-induced lung injury and its possible mechanism.

METHODSFifty male SD rats were equally randomized into phosgene exposure group, air control group, MT treatment group, dexamethasone (DX) treatment group, and negative control group. All groups except the air control group were exposed to 8.33 mg/L phosgene for 5 min, and the MT treatment group, DX treatment group, and negative control group were injected with MT (10 mg/kg), DX (2.5 mg/kg), and 1% ethanol saline (1 ml/kg), respectively, via the caudal vein 1 hour after exposure. The rats were sacrificed 6h later. Then, the wet/dry ratio of the lung, the total protein content and neutrophil count in bronchoalveolar lavage fluid (BALF), and the malonaldehyde (MDA) content and superoxide dismutase (SOD) and myeloperoxidase (MPO) activities in lung homogenate were measured; pathological observation was made on the lung tissue under an optical microscope; the protein expression of inducible nitric oxide synthase (iNOS) and NF-κB in the lung tissue was measured by Western blot.

RESULTSCompared with the air control group, the phosgene exposure group showed significantly increased wet/dry ratio of the lung and total protein content and neutrophil count in BALF (P < 0.01) as well as significantly increased MDA content and MPO activity in the lung tissue (P < 0.05). Compared with the phosgene exposure group, the MT treatment group showed significantly decreased MDA content and MPO activity and significantly increased SOD activity (P < 0.01), and the MT treatment group and DX treatment group showed significantly decreased protein expression of iNOS and NF-κB (P < 0.01).

CONCLUSIONMT has protective effect in phosgene-induced lung injury, and its protective mechanism may be associated with scavenging free radicals and inhibiting expression of iNOS and NF-κB.

Acute Lung Injury ; chemically induced ; metabolism ; prevention & control ; Animals ; Disease Models, Animal ; Male ; Malondialdehyde ; metabolism ; Melatonin ; pharmacology ; therapeutic use ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peroxidase ; metabolism ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism