Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.
Amino Acids ; analysis ; Animals ; Chickens ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Endothelium ; chemistry ; Gizzard, Avian ; chemistry

OBJECTIVETo analyze the relationship between the pulse pressure (PP) and atherosclerotic renal artery stenosis (ARAS) in middle-age and eldery paients with hyperensio, so as to provide the basis for the diagnosis and treatment of ARAS.

METHODSWe tetro spectively analyzed the data of 257 patients with hypertension undergoing renal arteriography at General Hospital of PLA between 2009 and 2014. Their mean age was (64.09 +/- 7.81) years. According the results of arteriography, they were divided into two groups: ARAS group (n = 145), and non-ARAS group (n = 112). Their PP and multinomial clinical indexes with ARAS were statistically analyzed.

RESULTSSingle factor correlation and Logisitic analysis showed that age, PP, serum creatinine (SCr) levels, and new onset of hypertension after 50 years of age were strongly positively related to ARAS. The age and PP and SCr was a team of continuous variable.

CONCLUSIONPP was the first new risk factor of ARAS in middile-aged and elderly patients with hypertension. It is suggested that PP may be a early predictive indicator and a new therapeutic target for ARAS.

Angiography ; Arteriosclerosis ; physiopathology ; Blood Pressure ; Humans ; Hypertension ; Middle Aged ; Renal Artery ; physiopathology ; Renal Artery Obstruction ; Risk Factors

OBJECTIVETo investigate the protective effect of overexpressed heme oxygenase-1 (HO-1) in hypoxia and the correlation between HO-1 overexpressoin and hypoxia inducible factor-1alpha (HIF-1alpha) in human hepatoma HepG2 cells.

METHODSThe expressions of HO-1 and HIF-1alpha mRNA as well as the protein were detected by RT-PCR and Western blotting, respectively. MTT assay was used to examine the relative cell survival rate. The total superoxide dismutase (SOD) activity was examined with a SOD kit.

RESULTSHypoxia induced overexpression of HO-1 gene in HepG2 cells at transcriptional and translational levels. The relative survival rate of HepG2 cells under hypoxia was significantly decreased after the HO-1 protein overexpression was inhibited by ZnPPIX (P < 0.01). The total SOD activity of cells was significantly increased after cells were treated by hypoxia for 16 hours (P < 0.05), while decreased significantly by HO-1 inhibitor ZnPPIX treatment (P < 0.01). HIF-1alpha was upregulated under hypoxia. In addition, the HO-1 overexpression under hypoxia was decreased by HIF-1alpha inhibitor, while the HIF-1alpha expression level under hypoxia was not significantly changed after HO-1 expression was inhibited by ZnPPIX.

CONCLUSIONThe overexpression of HO-1 in hypoxic HepG2 cells is HIF-1alpha-dependent or at least partly HIF-1alpha-dependent. The relative survival rate of hypoxic hepatoma cells was significantly decreased by HO-1 inhibitor treatment. The results of this study may offer new thought and drug target for the therapy of human hepatoma in the future.

Cell Hypoxia ; Cell Survival ; Heme Oxygenase-1 ; antagonists & inhibitors ; metabolism ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Protoporphyrins ; pharmacology ; RNA, Messenger ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo study and compare the effect of neutrophil elastase inhibitors (GW311616A and sivelestat) on the proliferation and apoptosis of U937 cells.

METHODSInhibitory effects of GW311616A and sivelestat on the proliferation of U937 cells were assayed by MTT assay. The morphologic changes of U937 cells were detected by transmission electron microscope, and apoptosis was observed by AnnexinV-FITC/PI staining. The changes of cell cycle and apoptosis were detected by flow cytometry. The expression of NE in U937 cells was observed by indirect immunofluorescence, the variations of content and activity of NE in U937 cells were measured through ELISA assay and colorimetric method.

RESULTSMTT showed that both NE inhibitors could inhibit the proliferation of U937 cells in a dose dependent manner. The IC50 of GW311616A and sivelestat were 150 and 214 μmol/L respectively. The inhibition effect of GW311616A was significantly higher than of sivelestat (P<0.01). Typical apoptosis morphological changes of U937 cells was observed through electron microscope. AnnexinV-FITC/PI staining showed that U937 cells could be induced to undergo apoptosis by the two inhibitors, the apoptosis ratio of 150μmol/L GW311616A group (13.60%) was significantly higher than that of 150μmol/L sivelestat group (3.69%)(P<0.01). The result of flow cytometry indicated that the apoptosis ratio of 150 μmol/L GW311616A group was 14.61%, U937 cell cycle was mainly blocked in G2/M phase; meanwhile 150 μmol/L sivelestat group as 4.25% with cell cycle in S phase. The fluorescence intensity of GW311616A group obviously decreased than of sivelestat group. And the two inhibitors could reduce the content and activity of NE in U937 cells, but the effect of GW311616A was significantly higher than of sivelestat (P<0.01).

CONCLUSIONGW311616A and sivelestat could inhibit the proliferation and cause apoptosis of U937 cells. Furthermore, GW311616A was more effective and harmful to cells than sivelestat.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Glycine ; analogs & derivatives ; pharmacology ; Humans ; Leukocyte Elastase ; antagonists & inhibitors ; Piperidines ; pharmacology ; Proteinase Inhibitory Proteins, Secretory ; pharmacology ; Sulfonamides ; pharmacology ; U937 Cells

OBJECTIVETo study the applied anatomy of the vascular and muscular innervations related to vascularized fibular grafts.

METHODSThirty-four cadaveric lower extremities were dissected for this study. The observations included fibular length, fibular nutrient artery, arcuate arteries, and innervation of fibular muscles. The fibulas were averagely divided into four segments and the locations of relevant vessels and nerves were ascertained.

RESULTSAll specimens had 1 fibular nutrient artery and 4-9 arcuate arteries except 1 specimen which had only 1 arcuate artery. The fibular nutrient artery and the first three arcuate arteries were constantly located between the distal half of the 1/4 segment and 2/4 segment of the fibula. The muscular branch of the superficial peroneal nerve passed through the surface of the periosteum in the 2/4 segment of the fibula.

CONCLUSIONSThe most proximal osteotomy point locates at the midpoint of the 1/4 segment by which it ensure the maximal potential for preserving the nutrient vessels. The muscular branch of the superficial peroneal nerve is fragile to injury at the 2/4 segment of the fibula.

Cadaver ; Female ; Fibula ; blood supply ; innervation ; Humans ; Male

Action Potentials ; Animals ; Electromyography ; Environmental Exposure ; Hypothermia ; Rats ; Rats, Wistar ; Seawater ; Thigh ; physiology ; physiopathology

Adenosine Triphosphatases ; genetics ; Adult ; Cation Transport Proteins ; genetics ; Copper-transporting ATPases ; Genotype ; Hepatolenticular Degeneration ; genetics ; Humans ; Liver ; metabolism ; Male ; RNA, Messenger ; analysis

BACKGROUNDIt is difficult and time-consuming for carrying out conventional hand-sewn bilioenteric anastomosis, especially for small bile duct anastomosis and laparoscopic procedure. In order to simplify it, we have developed a novel procedure of sutureless bilioenteric anastomosis with an intraluminal degradable stent. This study aimed to evaluate the feasibility and safety of this technique with cholangioduodenostomy in dog model.

METHODSA patent intraluminal degradable stent tube for sutureless choledochoduodenostomy in dog model was made with polylactic acid in diameter of 3 mm or 4 mm. Thirty-eight dogs were randomly divided into to a stent group (SG, n = 20) and a control group (CG, n = 18). Dogs in the SG underwent sutureless choledochoduodenostomy with intraluminal stent, while the CG underwent conventional choledochoduodenostomy (single layer discontinuous anastomosis with absorbable suture). Dogs of each group were divided into 4 subgroups according to time of death (1, 3, 6, and 12 months postoperatively) to evaluate the healing of anastomosis. Operation time, intraoperative tolerance pressure of anastomosis, rate of postoperative bile leakage, bursting pressure of anastomosis were compared between the two groups. Anastomosis tissue was observed afterwards by pathology evaluation, hydroxyproline content, serum bilirubin, liver enzyme level and magnetic resonance cholangio-pancreatography (MRCP) to assess the stricture.

RESULTSAll procedures were completed successfully. The surgical time of the SG was significantly less than the CG (SG: (19.2 ± 4.3) minutes, vs. CG: (29.2 ± 7.1) minutes, P = 0.000). One bile leakage was occurred in either group. No significant difference of intraoperative tolerance pressure of anastomosis, rate of bile leakage and postoperative bursting pressure of anastomosis, anastomotic stricture, hydroxyproline content, serum bilirubin and liver enzyme level was found between the two groups. MRCP showed no anastomosis stricture and obstruction during months of follow-up.

CONCLUSIONThe technique of sutureless choledochoduodenostomy with a degradable intraluminal stent is feasible and a safe procedure in this dog model.

Animals ; Choledochostomy ; methods ; Dogs ; Female ; Male ; Stents

OBJECTIVE: To describe the imaging features of pelvic solitary plasmacytoma and to correlate them with the pathologic grade. MATERIALS AND METHODS: A retrospective study was performed on the imaging features of 10 patients with a histological diagnosis of pelvic solitary plasmacytoma. The imaging studies were assessed for bone expansion, cortical destruction, signal intensity/density of soft tissue mass and enhancement manifestations, which were then correlated to the pathologic grade. RESULTS: The imaging features of pelvic solitary plasmacytoma revealed 3 different types: multilocular type (n = 5), unilocular type (n = 2) and complete osteolytic destruction type (n = 3) on computed tomography and MRI. Pathologically, the tumors were classified into low, intermediate and high grades. Features such as multilocular change, perilesional osteosclerosis, slight expansion, local bone cortex disruptions and masses inside bone destruction, often suggest a low-grade solitary plasmacytoma; complete osteolytic destruction, huge soft tissue mass, and osseous defects imply a higher pathologic grade. CONCLUSION: Pelvic solitary plasmacytoma has various imaging manifestations, while a slight expansile osteolytic feature with multilocular change or homogeneous enhancement highly suggests its diagnosis. The distinctive imaging features of pelvic solitary plasmacytoma are well correlated to the pathologic grade.
Adult ; Aged ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm Grading ; Pelvic Neoplasms/*pathology/radiography ; Plasmacytoma/*pathology/radiography ; Retrospective Studies ; Tomography, X-Ray Computed

The cDNA fragments of interest were amplified using Sj lambda ZipLox library as the templates by PCR and then cloned into a eukaryotic expression vector p-CMV-GH; A small number of DNA fragments inserted in the recombinants was identified by restriction cleavage, EST sequencing and bioinformatical analysis; mice were injected intramuscularly with the expression library (L-CMV-SjR) or sublibraries(L-CMV-SjR1, L-CMV-SjR2 and L-CMV-SjR3), immunized mice were challenged with Schistosoma japonicum cercariae on day 35, the levels of IgG antibodies in sera from the immunized mice were detected by ELISA. The results demonstrated that a partial cDNA expression library of S.j, with approximately 10(5) transformants, was constructed, most of the recombinants contained the insert DNA fragments of interest, and these fragments had the features of protein-coding sequences for Schistosome. There were no significant differences for the levels of IgG antibodies in sera from all of the immunized groups. Mice immunized with L-CMV-SjR, L-CMV-SjR1 and L-CMV-SjR2 developed significant protective effect against Sj infection compared to control mice injected with the empty plasmid, the rate of worm reduction was about 30%.
Animals ; Antibodies, Helminth ; blood ; immunology ; Antigens, Helminth ; genetics ; immunology ; DNA, Complementary ; Disease Models, Animal ; Gene Expression ; Gene Library ; Mice ; Schistosoma japonicum ; genetics ; immunology ; Schistosomiasis japonica ; prevention & control ; Vaccination ; methods ; Vaccines, DNA ; genetics ; immunology