The effect of C-terminal region residues on the substrate specificity of a novel cyclic imide hydrolase (CIH), a recombinant cyclic imide hydrolase (CIH293), and its mutants deleted or substituted at C-terminus (CIH291, CIH290, KK292-293EE) was reported. The substrate specificity and kinetic parameters of the mutants were analyzed by both the spectrophotometric assay and high-performance liquid chromatography. Results show that the substrate specificity of mutants was not obviously changed, but slightly low for the affinity between the substrate and enzyme, compared with the wild-type enzyme, CIH293. In conclusion, the last three residues of CIH293 play an important role for the enzyme activity.

Objective To study adrenomedullin (AM) mRNA and protein expression level in myocardium of autoimmune myocarditis animal models induced by immunization of mice with lactobacillus casei cell wall element(LCWE). Methods Forty-five Balb/c male mice were randomly divided into experimental group (n = 30) and control group (n = 15), which were intraperitoneally injected with LCWE and phosphate buffered solution(PBS) at day 0,3,5 and 10,respectively. Sera and myocardium samples were gained 14,21 and 28 days after the first immunization. AM expression levels were determined by semiquantitative reverse transcriptase-polymerase chain reaction(RT- PCR) and immunchistochemistry,and mycardial histopathological lesions were observed. The anti- myosin antibodies in different stages were examined by an ELISA. Results There were myocardial necrosis or inflammatory infiltration in the experimental group, but myocardial lesions were not found in the control group. Anti - myosin antibodies were detected in sera of experimental mice,but not in control group. Immunchistochemistry findings demonstrated that AM expression level was higher in the experimental group than in the control group( P

Objective To evaluate tension-free hernia Onlay repair with premuscular positioning of the prosthesis for the treatment of ineisional henia of the abdominal wall.Methods In this study 126 patients with incisional henia were treated with a tension-free manner of hernia repair by using synthetic material ONLAY between September 1999 and June 2007.Results All operations were successful.There was no hospital death or severe postoperative complications.The average age was 58.5 years old ranging from 28 to 89.There were 67 patients in which the abdominal defect ranged from 5～10 cm,and 59 patients with abdominal defect≥10 cm.The mean operating time was 95(70～120)min,and the average intraoperative blood loss was 80 ml(60～250 ml).The mean postoperative hospitalization was 14.5 days(10～28 d). Patients were followed-up from 3 to 96 months,and 3 patients suffered from hernia recurrence(2.38%). Conclusions The ONLAY repair of ineisional hernia of the abdominal wall with synthetic material mesh was a safe procedure,especially for those with large abdominal wall defects.

Total RNA was extracted from infected papaya (Carica papaya L.) leaves in Hainan Province, and the full-length sequences of papaya ringspot virus were amplified by RT-PCR and RACE, and its complete genomic sequence was assembled, named Hainan-P isolate. The RNA genome sequence of Hainan-P isolate was 10323 nucleotides (nts)in length,excluding the 3'-terminal poly(A) tail. And it was composed of a single open reading frame encoding a polyprotein of 3343 amino acids.. The result of homology analysis with twelve GenBank PRSV isolates showed that the polyprotein identity of Hainan-P ranged from 89. 8% to 93.2%, that was higher than the complete nt homology of 82.3% to 89.1%. The P1 amino acid was the least conserved (sharing homology only between 65.4% and 80.1%), whereas HC-Pro, CI and CP were the most conserved. Phylogenetic tree were constructed by the Neighbor-joining method in MEGA 3.1, which showed that PRSV isolates were obviously relevant to geographical origin, and it was impossible to delineate host-specific (P type and W type)evolution.
Carica ; virology ; Genome, Viral ; Phylogeny ; Potyvirus ; classification ; genetics

OBJECTIVETo study hepatitis B virus (HBV) genotype and subtype distribution and its clinical significance in HBV-infected patients.

METHODSWe used type/subtype-specific primers and PCR to detect HBV genotype and subtype of 445 HBV-infected patients from Beijing, Changchun, Hanchuan Shenzhen, Qingyuan and Nanjing, including 7 acute hepatitis (AH), 36 asymptomatic HBV carriers (ASC), 352 chronic hepatitis (CH), 28 liver cirrhosis (LC), and 22 hepatocellular carcinoma (HCC) cases. Genotyping results were confirmed by PCR product sequencing.

RESULTSAmong 445 HBV-infected patients, the proportions of genotype B, C, and B/C were 32.6% (145/445), 53.7% (239/445), and 13.7% (61/445), respectively. In genotype C, 13 (5.4%) were subtype C1, 135 (56.5%) were subtype C2, and the remaining 91 (38.1%) were neither C1 nor C2. In genotype B, 100 (69.0%) were subtype Ba, 25 (17.2%) subtype Bj, and the other 20 (13.8%) were neither Ba nor Bj. In genotype B/C, 15 (24.6%) were Ba/C2, 8 (13.1%) Bj/C2, 6 (9.8%) Ba/C1, 3 (4.9%) Bj/C1, 11 (18.0%) Ba/neither C1 nor C2, 7 (11.5%) Bj/neither C1 nor C2, and 6 (9.8%) neither Ba nor Bj/neither C1 nor C2, 2 (3.3%) neither Ba nor Bj/C1, 3 (4.9%) neither Ba nor Bj/C2. The HBV genotype and subtype distribution we found exhibited significant differences in the various clinical types of HBV infection tested, and showed that genotype C was predominant among patients with liver cirrhosis (78.6%) and hepatocellular carcinoma (86.4%) while genotype B was predominant in asymptomatic carriers (72.2%). In addition, genotype and subtype distribution showed no significant differences between male and female patients, but genotype and subtype distribution showed significant differences in patients positive or negative with HBeAg.

CONCLUSIONSubtypes Ba and C2 are predominant in patients with hepatitis B from these 6 cities, and genotype C may be associated with the development of liver cirrhosis and hepatocellular carcinoma.

Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; virology ; Polymerase Chain Reaction

OBJECTIVETo investigate the optimal time and method of the early repair of the full layer eyelid defect caused by chemical burn.

METHODSFree nasal septum mucosal cartilage flap with muscle flap, skin grafting, or skin flap were performed in 18 cases (19 eyelids) with chemical burn within 4 postburn weeks. Eyelid reconstruction and corneal transplantation were performed at the same time in 4 patients.

RESULTSAll the reconstructed eyelids and transplanted cornea survived. The incidence of severe complications, such as exposure keratitis, corneal ulcer and eyeball perforation decreased.

CONCLUSIONFull layer eyelid defect caused by chemical burn should receive early reconstruction and repair, including timely reconstruction of eyelid for the sake of protecting the eyesight and of alleviating the inflammatory reactions, and the corneal transplantation should be done at the same time to avoid corneal perforation. Nasal septum mucosal cartilage flap could be ideal for the eyelid reconstruction.

Adult ; Burns, Chemical ; surgery ; Corneal Transplantation ; Eyelids ; injuries ; surgery ; Female ; Humans ; Male ; Middle Aged

Objective To establish a QAMS method for content determination of six compositions (chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin) from Lonicerae Japonicae Caulis; To verify the feasibility and applicability of this method in quality control of Lonicerae Japonicae Caulis. Methods Chlorogenic acid was set as internal reference substance. The HPLC analysis was performed on a Waters Symmetry C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase consisted of acetonitrile and 0.4% phosphoric acid solution in gradient elution manner at a flow rate of 1 mL/min. The column temperature was maintained at 35 ℃, and the detection wavelength was set at 327 nm for chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and 236 nm for loganin. Results The relative correction factors of caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin were established; there was no obvious difference between calculated value of QAMS and measured value of external standard method. Conclusion The quality control mode of QAMS can be used for multi-index synchronization quality evaluation of the six compositions from Lonicerae Japonicae Caulis.

Cadherins ; metabolism ; Female ; Humans ; MCF-7 Cells ; Neoplastic Stem Cells ; metabolism ; Receptor, Notch1 ; metabolism ; Receptors, CXCR4 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism

BACKGROUNDPulmonary embolism (PE) is often mistaken as acute coronary syndromes (ACS) because of the considerable overlap in their clinical features. We evaluated the factors causing misdiagnosis of PE as ACS and factors that differentiate PE from ACS to improve the diagnosis efficacy of PE.

METHODSThe medical records of 22 consecutive PE patients, between 2001 and 2010, who were initially suspected of ACS were retrieved. ACS was ruled out by coronary artery angiography before a definite diagnosis of PE was given. Twenty-two contemporary cases of ACS matched by age and sex were recruited as controls. Clinical manifestations, electrocardiograms (ECG), and biomarkers of these patients were reviewed retrospectively. The factors causing misdiagnosis of PE as ACS and factors differentiating PE from ACS were evaluated.

RESULTSWe found two leading causes of misdiagnosis of PE as ACS. One is that PE can resemble ACS in several clinical aspects (symptoms and signs, ECG findings, plasma cardiac troponin I, and D-dimer). The other is the insufficient recognition of PE by clinicians. Risk factors for venous thromboembolism (VTE), especially deep venous thrombosis (DVT), together with signs of PE, such as unexplained dyspnea or hypoxemia, and right ventricular pressure overload on ECGs are valuable in differentiating the two diseases.

CONCLUSIONSDifferentiation between PE and ACS is sometimes challenging. Adequate awareness of the risk factors for VTE and the signs of PE are crucial in the diagnosis of PE.

Acute Coronary Syndrome ; diagnostic imaging ; Adult ; Aged ; Coronary Angiography ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Embolism ; diagnostic imaging ; Retrospective Studies

OBJECTIVETo observe the effects of different sized titanium dioxide on reactive oxygen species (ROS) levels.

METHODSForty-eight healthy mice were divided into 3 groups randomly: control group (distilled water), 50 nm TiO₂ (5 g/kg) group and 120 nm TiO₂ (5 g/kg) group. The mice were exposed to distilled water or TiO₂ by a syringe via gastrointestinal tract in a minute. One week later, the mice were sacrificed and the liver, kidney and brain (cortex, hippocampus) were collected. The contents of Ti in the above tissues of mice were measured by ICP-MS. The ROS levels in the tissues were measured by flow cytometry.

RESULTSThere was no significant difference in organ coefficient between two exposure groups and control group (P > 0.05). The Ti contents in liver, kidney, cortex and hippocampus of two exposure groups were higher than those of control group (P < 0.05); The Ti contents of above four tissues in 50 nm TiO₂ group were higher than those in 120 nm group (P < 0.05). The ROS levels in liver, kidney and cortex cells of two exposure groups (273.2 ± 32.5, 160.2 ± 28.5, 74.9 ± 8.9; 159.4 ± 15.9, 64.4 ± 7.5, 41.2 ± 5.6) significantly increased, as compared with control group (74.9 ± 6.4, 24.9 ± 2.8, 32.8 ± 3.1) (P < 0.05). The ROS levels in hippocampus cells in 50 nm TiO₂ group were significantly higher than those in control group (P < 0.05). The ROS levels of above four tissues in 50 nm group were significantly higher than those in 120 nm TiO₂ group (P < 0.05).

CONCLUSIONAfter mice were exposed to 50 nm and 120 nm TiO₂ particles via gastrointestinal tract, Ti could be distributed into the liver, kidney and the brain tissues in mice, resulting in the enhanced ROS levels in liver, kidney, cortex and hippocampus cells. Moreover, the ROS levels induced by 50 nm TiO₂ particles are significantly higher than those induced by 120 nm TiO₂ particles.

Animals ; Lipid Peroxidation ; drug effects ; Male ; Mice ; Mice, Inbred Strains ; Particle Size ; Reactive Oxygen Species ; metabolism ; Titanium ; chemistry ; toxicity