In order to obtain the bifunctional chimeric molecule of single-chain urokinase-type plasminogen activator (scu-PA) which can inhibit platelet aggregation, PRGDWR peptide was inserted into the site between Gly 118 and Leu119 (called insertion mutant B, InB). The recombinant gene of InB was expressed by Pichia pastoris. The secreted protein was purified by metal chelate affinity and strong cation exchange chromatography. The amidolytic ability of mutant InB is 5 900 IU/mg, the kinetic constants is: KInB m,plg=56.8 μmol*L-1，kInBcat,plg=0.33 s- 1. The kinetic constants of plasminogen activation reaction is: KInB m,plg=0.397 μmol*L-1，kInBcat,plg=0.0164 s-1. Fibrin inhibit the catalytiv ability of InB during plasminogen activatio n, the influence factor is 0.463(means InB remain 46.3% of the catalytic abili ty when fibrin was involved in the reaction system). The mutant not only has alm ost the same catalytic ability as wild type scu-PA, but also has strong ability of anti-platelet aggregation(compared with scu-PA), IC50 of InB is 12.7 μmol*L-1.

Because the influence of fibrin on the reaction of plasm inogen activation by various plasminogen activators is different, the kinetic co nstant of the reaction of plasminogen activation catalyzed by InB with and witho ut fibrin were detected. The result is: Kfibrinm＝4.2 μmol*L -1，greater than the normal Km=0.379 μmol*L-1; kfib rincat＝0.107 s-1，greater than the normal kcat=0.0165 s-1. The results suggest that existence of fibrin in the reaction system of plasminogen activation depress the affinity between InB and plasminog en, but accelerates the hydrolysis of plasminogen by InB. The count up effect is inhibition.

Objectives To discuss the CT features of primary tuberculosis in adults Methods CT images of 39 adult patients with primary tuberculosis were retrospectively analyzed and 30 cases were also examined after injection of contrast materials Results 39 patients had both mediastinal tuberculous lymphadenitis and parenchymal infiltration There were 25 cases (64 1%) with primary lesions involving the right lung and 14 cases (35 9%) in left side 28 cases(71 8%) appeared as patchy shadows,nodules,and lobular consolidation; 11 cases (28 2%) were with segmental or lobar consolidation In 30 cases with contrast enhanced CT,the mediastinal tuberculous lymphadenitis appeared as homogenous enhancement in 10 cases, irregular enhancement in 18 cases,central low attenuation and peripheral rim enhancement in 20 cases 14 cases showed that the nodes were confluenced with inhomogenous enhancement There were 26 cases with acute bronchogenic spread tuberculosis Conclusion The parenchymal infiltration and mediastinal tuberculous lymphadenitis are the basic features in adult primary tuberculosis Acute bronchogenic spread into other fields and the features of node enhancement after injection of contrast materials are very important findings in the diagnosis of primary tuberculosis

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

OBJECTIVETo investigate the effects of carbaryl on serum steroid hormone and function of antioxidant system in female Sprague Dawley rats.

METHODSCarbaryl was administrated to the adult female rats at doses of 0, 1.028, 5.140 and 25.704 mg x kg(-1) x d(-1) for 30 d. Vaginal smears of rats were taken to determine estrous cycle. Serum 17beta-estradiol (E(2)) and progesterone (P(4)) concentrations were measured by radioimmunoassay. The activities of superoxide dismutase (SOD) and glutathione-s-transferase (GST), and the levels of malondialdehyde (MDA) and glutathione (GSH) were measured by spectrophotometry.

RESULTSThe number of estrous cycle in exposed groups were obviously lower than in control group (P < 0.05, P < 0.01). Body weight gain in high dose group (25.704 mg x kg(-1) x d(-1)) was significantly lower than that in control. Meanwhile, the organ coefficient of ovary and uterus declined in a dose-dependent manner. Serum E(2) level [(19.93 +/- 2.21) nmol/L] in 25.704 mg group was lower than in control group [(28.76 +/- 6.12) nmol/L, P < 0.05], and P(4) level (1.21 +/- 0.40) nmol/L in 1.028 mg group was higher than that in control group [(0.63 +/- 0.39) nmol/L, P < 0.05]. The activity of SOD first reduced then rose in ovary, and first rose then reduced in serum. The contents of MDA increased in ovary, while decreased in the serum. GSH contents and GST activity increased in ovary, while in serum GSH contents decreased and GST activity first increased then decreased.

CONCLUSIONCarbaryl could disrupt estrous cycle and affect serum steroid hormone, and the function of antioxidant system in female SD rats.

Animals ; Antioxidants ; metabolism ; Carbaryl ; toxicity ; Estradiol ; blood ; Female ; Glutathione ; blood ; Glutathione Transferase ; blood ; Malondialdehyde ; blood ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

Viral hepatitis was the most common infectious disease in china. But the diagnosis and treatment were varied because the viral hepatitis patients were hospitalized in different kinds of hospital such as infectious disease hospital, general hospital and Chinese medical hospital. It was necessary to know clinical characters and information of viral hepatitis patients in different hospitals. The general information, subtype distribution, prognosis, complication, medication and relations of onset with solar term from 41 180 viral hepatitis patients based on HIS data were analyzed. It was found that the age of patients between 18 to 59 years old was most; most patients were males. The national basic medical insurance was the most type of payment. The outcome of viral hepatitis in the youth and female were better than that in the old and male. Acute hepatitis was easer to restore than chronic hepatitis. Liver cirrhosis and hepatocellular carcinoma were the two most complications. The peak of onset was during summer solstice, slight heat and great heat. The most common Chinese medicine was Diammonium glycyrrhizinate and the most common western medicine was reduced glutathione. The combination of D. glycyrrhizinate with reduced glutathione, polyene phosphatidylcholine and thymosin was the main pattern. But It was not knew if the combination of western and Chinese medicine was the most effective therapy to protect liver function. It was necessary to take deeply research of the relationship between the combination therapy and their effectiveness.
Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Glutathione ; therapeutic use ; Glycyrrhizic Acid ; therapeutic use ; Hepatitis, Viral, Human ; drug therapy ; Hospitals ; Humans ; Male ; Middle Aged ; Young Adult

Objective To establish high-performance liquid chromatography with electrochemical detector(HPLC-ECD) method for the determination for metanephrine and normetanephrine in 24 h urine, and provide a superior test for the diagnostic of pheochromocytomas over plasma/urine catecholamine.Methods MCX solid-phase cartridge was used for extraction of metanephrine and normetanephrine,HPLC-ECD was used for their measurements.The intra-assay CVs,interassay CVs and recoveries of metanephrine and normetanephrine were also calculated.104 hypertensive patients without pheochromocytomas and 5 pheochromocytomas patients were selected in this study.The concentrations of metanephrine and normetanephrine were compared with the plasma and 24h urinary catecholamines concentrations.Results The intra-assay CV,inter-assay CV and recovery of metanephrine were 5.9%, 7.5%,91.1% respectively;the intra-assay CV,inter-assay CV and recovery of normetanephrine were 6.3%,6.6%,88.5%,respectively.The MN,NMN,plasma CA and urine CA of all pheochromocytomas patients were positive.MN and NMN were negative in controls,while plasma CA and urine CA are false positive in 15 patients and 14 patients in controls,respectively.Conclusions The study establish a fast and accurate method for quantification of metanephrine and normetanephrine in 24 h urine by HPLC-ECD.These findings also prove that it is the best biochemical assays for pheochromocytomas at present.

Stones in the seminal vesicles are extremely rare. We present a 62-year-old patient with a stone within a seminal vesicle cyst, who was cured by laparoscopic treatment. The operative time was 80 min, and the estimated blood loss was 90 mL. Scanning electron microscope examination of the stone showed a compact crystal image externally and sparse spherical crystal structure in kernel. Composition of the stone was calcium fluorophosphate on X-ray diffractometer. The follow-up time was 15 months with no recurrence of cyst or stone. To our knowledge, this case is the first to describe laparoscopic removal of a stone within a seminal vesicle cyst, and the first to describe calcium fluorophosphate as the composition of seminal vesicle stones.
Calculi ; surgery ; Cysts ; surgery ; Genital Diseases, Male ; surgery ; Humans ; Laparoscopy ; Male ; Microscopy, Electron, Scanning ; Middle Aged ; Seminal Vesicles

OBJECTIVETo investigate the effcacy of the staple fixation for the treatment of hamate metacarpal joint injury.

METHODSFrom May 2009 to November 2012,16 patients with hamate metacarpal joint injury were treated with staple fixation including 10 males and 6 females with an average age of 33.6 years old ranging from 21 to 57 years. Among them, 11 cases were on the fourth or fifth metacarpal base dislocation without fractures, 5 cases were the fourth or fifth metacarpal base dislocation with avulsion fractures of the back of hamatum. Regular X-ray review was used to observe the fracture healing, joint replacement and position of staple fixation. The function of carpometacarpal joint and metacarpophalangeal joint were evaluated according to ASIA (TAM) system evaluation method.

RESULTSAll incision were healed well with no infection. All patients were followed up from 16 to 24 months with an average of (10.0 +/- 2.7) months. No dislocation recurred, the position of internal fixator was good,no broken nail and screw withdrawal were occurred. Five patients with avulsion fracture of the back of hamatum achieved bone healing. The function of carpometacarpal joint and metacarpophalangeal was excellent in 10 cases,good in 5 cases, moderate in 1 case.

CONCLUSIONThe application of the staple for the treatment of hamatometacarpal joint injury has the advantages of simple operation, small trauma, reliable fixation, early postoperative function exercise and other advantages, which is the ideal operation mode for hamatometacarpal joint injury.

Adolescent ; Adult ; Carpal Joints ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Fractures, Bone ; surgery ; Hamate Bone ; injuries ; surgery ; Humans ; Male ; Metacarpal Bones ; injuries ; surgery ; Metacarpophalangeal Joint ; injuries ; surgery ; Middle Aged ; Sutures ; utilization ; Young Adult