The involvement of plasma kallikrein-kinin system in some pathological states such as bronchial asthma and coughing induced by captopril has been suggested, LTC, and LTD, have been well reported to have a potent activity of bronchoconstriction and mucous secretion. The author tried to confirm whether leukotrienes increase glandular kallikrein activity in bronchial wash or not, and if it is true, to investigate the possible mechanism of leukotrienes-induced increase of glandular kallikrein activity. Bronchial wash was collected from excised lungs of guinea pig, and incubated with synthetic substrate (Pro-Phe-Arg-MCA), and released amount of AMC was measured by fluorescence spectrophotometer as the amidase activity (glandular kallikrein activity). In order to confirm that glandular kallikrein can release kinin, bronchial wash was also incubated with purified LMW- kininogen in the presence of kininase inhibitor (o-phenanthroline), and the amount of kinin was measured by enzyme immunoassay as the kinin releasing activity of glandular kallikrein. The results were as follows: 1) The glandular kallikrein activity (control: 2.98 x 10(-11) mole/min/ml wash) was increased by pilocarpine (12-120 umole/kg, i.v.), and the increased glandular kallikrein activity by pilocarpine (41 nmole/kg, i.v.) was inhibited by atropine (4-43 nmole/kg, i.v.). 2) LTC(4) (1-10 nmole/kg, i.v.) or LTD(4) (1-10 nmole/kg, i.v.) increased the glandular kallikrein activity, but the potency of LTD, was about 1/3 of that of LTC,. The increased glandular kallikrein activity by LTC, (3 nmole/kg, i.v.) was inhibited by ONO-1078 (20-203 nmole/kg, i.p.). 3) The increased glandular kallikrein activity by LTC, (3nmole/kg, i.v.) was inhibited by in- domethacin (8-83 nmole/kg, i.p.), OKY-046 (11-113 nmole/kg, i.v.), atropine (4-43 nmole/kg, i.v.), scopolamine (9-88 umole/kg, i.v.) and Thi-D-Phe-BK (100-1000 nmole/kg, i.v.). 4) STA, (6-60pmole/kg, i.v.) increased the glandular kallikrein activity, and the increased glandular kallikrein activity by STA, (20 pmole/kg, i.v.) was inhibited by atropine (14 umole/kg, i.v.) (p < 0. 001). 5) The increased glandular kallikrein activity by pilocarpine (41 urnole/kg, i.v.) was not inhibited by indomethacin (83 umole/kg, i.p.), but it was inhibited by Thi-D-Phe-BK (300 nmole/kg, i.v.) (p< 0. 001). 6) Bradykinin (3-30 nmole/kg, i.v.) increased the glandular kallikrein activity, and the increased glandular kallikrein activity by bradykinin (30 nmole/kg, i.v.) was inhibited by indomethacin (83 umole/kg, i.p.) or atropine (14 umole/kg, i.v.) (p<0.001). 7) The kinin releasing activity by glandular kallikrein in bronchial wash was 24 x 10 " mole/min/ ml wash in control group, and it was markedly increased in pilocarpine-pretreated group (124 x 10 mole/min/ml wash) (p<0.05), and in LTC4-pretreated group (164x10 mole/min/ml wash) (p<0. 01). The increased kinin releasing activity by LTC, (3 nmole/kg, i.v.) was completely inhibited by aprotinin (5000IU/ml, i.v.). I could conclude that i ) LTC, and LTD, increased glandular kallikrein activity in bronchial wash of guinea pig, ii) the most possible mechanism of LTC,-induced increase of glandular kallikrein activity in bronchial wash may be as follows; LTC,TXA,aeetyleholineinerease of glandular kallikrein acitivity, and iii) kinin released by glandular kallikrein may enhance the action of TXA, or acetylcholine to increase the glandular kallikrein activity.
Acetylcholine ; Animals ; Aprotinin ; Asthma ; Atropine ; Bradykinin ; Bronchoconstriction ; Captopril ; Cough ; Fluorescence ; Guinea Pigs* ; Guinea* ; Immunoenzyme Techniques ; Indomethacin ; Kallikrein-Kinin System ; Kallikreins ; Kininogens ; Leukotriene C4* ; Leukotrienes ; Lung ; Pilocarpine ; Plasma ; Scopolamine Hydrobromide ; Tissue Kallikreins*

OBJECTIVETo find sensitive and specific micro-metastic markers for prostate cancer.

METHODSUsing nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.

RESULTSThe expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.

CONCLUSIONPSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

Aged ; Antigens, Surface ; blood ; Biomarkers, Tumor ; blood ; Glutamate Carboxypeptidase II ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatic Neoplasms ; blood ; pathology ; Tissue Kallikreins ; blood

Aged ; Humans ; Kallikreins ; analysis ; Male ; Middle Aged ; Prognosis ; Prostate-Specific Antigen ; analysis ; Prostatic Neoplasms ; diagnosis

OBJECTIVE: To determine the predictive accuracy of the combined panels of serum human tissue kallikreins (hKs) and CA-125 for the detection of epithelial ovarian cancer. METHODS: Serum specimens collected from 5 Indonesian centers and 1 Vietnamese center were analyzed for CA-125, hK6, and hK10 levels. A total of 375 specimens from patients presenting with ovarian tumors, which include 156 benign cysts, 172 epithelial ovarian cancers (stage I/II, n=72; stage III/IV, n=100), 36 germ cell tumors and 11 borderline tumors, were included in the study analysis. Receiver operating characteristic analysis were performed to determine the cutoffs for age, CA-125, hK6, and hK10. Sensitivity, specificity, negative, and positive predictive values were determined for various combinations of the biomarkers. RESULTS: The levels of hK6 and hK10 were significantly elevated in ovarian cancer cases compared to benign cysts. Combination of 3 markers, age/CA-125/hk6 or CA-125/hk6/hk10, showed improved specificity (100%) and positive predictive value (100%) for prediction of ovarian cancer, when compared to the performance of single markers having 80-92% specificity and 74-87% positive predictive value. Four-marker combination, age/CA-125/hK6/hK10 also showed 100% specificity and 100% positive predictive value, although it demonstrated low sensitivity (11.9%) and negative predictive value (52.8%). CONCLUSION: The combination of human tissue kallikreins and CA-125 showed potential for improving prediction of epithelial ovarian cancer in patients presenting with ovarian tumors.
Asian Continental Ancestry Group ; Humans ; Neoplasms, Germ Cell and Embryonal ; Neoplasms, Glandular and Epithelial ; Ovarian Neoplasms ; ROC Curve ; Sensitivity and Specificity ; Tissue Kallikreins ; Biomarkers, Tumor

In 2011, a number of studies shed further light on the understanding of stroke etiologies and mechanism. Regarding eating habit and a risk of stroke, three studies from Swedish cohort showed independent relations of chocolate, red meat, and coffee consumption with a risk of stroke. In relation to biomarkers, plasma tissue Kallikrein and resistin levels have been shown to be associated with a risk of stroke. In addition, with regard to infection and stroke, one study reported an increasing incidence of HIV infection in stroke patients and another study demonstrated that new-onset atrial fibrillation among patients with severe sepsis is associated with an increased risk of stroke. Finally, European consortium of cervical artery dissection reported risk factors for cervical artery dissection and differences between carotid and verterbral artery dissections.
Arteries ; Atrial Fibrillation ; Biomarkers ; Cacao ; Coffee ; Cohort Studies ; Eating ; HIV Infections ; Humans ; Incidence ; Light ; Meat ; Plasma ; Resistin ; Risk Factors ; Sepsis ; Stroke ; Tissue Kallikreins

OBJECTIVETo investigate the effects of human tissue kallikrein 1(Ad-hKLK1) gene delivery on the neointima formation in carotid arteries of spontaneously hypertensive rats (SHRs).

METHODSCarotid artery restenosis was induced in male SHR rats by balloon-injury. Rats were randomly assigned into 4 groups: Sham-operated (n = 6); Angioplasty (phosphate buffered solution 50 microl, n = 8); Vector virus (control virus, 1 x 10(9) IU in 50 microl, n = 8) and Ad-hKLK1(Ad-hKLK1, 1 x 10(9) IU in 50 microl, n = 8). Rats were sacrificed 4 weeks later. The wall-to-lumen area ratio and intima/media ratio in carotid artery were assessed by image analysis in HE stained sections. The mRNA bradykinin receptor (B1R and B2R) expressions were detected by RT-PCR. The protein expression of the cycle-independent kinase inhibitors p27Kip1 and p2lCip1 were determined by Western blot analysis.

RESULTSWall-to-lumen area ratio reduced 35.6% and intima/media ratio reduced 38.8%in Ad-hKLK1 treated SHRs compared to angioplasty group (all P < 0.001). The expression of p27Kip1 and p2lCip1 increased significantly in Ad-hKLK1 treated SHRs compared with angioplasty rats (all P < 0.001). The mRNA expression of B2R was significantly upregulated in angioplasty rats compared with sham-operated rats (P < 0.05) while mRNA expression of B1R was similar between the 2 groups.

CONCLUSIONhKLK1 gene delivery may effectively reduce neointimal formation via downregulating bradykinin B2R and up-regulating the expressions of p27Kip1, p2lCip1 signaling pathways in carotid arteries of SHRs after balloon injury.

Angioplasty, Balloon ; adverse effects ; Animals ; Carotid Artery, Common ; pathology ; Gene Transfer Techniques ; Humans ; Male ; Neointima ; etiology ; Rats ; Rats, Inbred SHR ; Tissue Kallikreins ; genetics

OBJECTIVETo evaluate the effects of urinary kallidinogenase on subarachnoid hemorrhage (SAH) in rabbits.

METHODSRabbits symptomatic cerebral vasospasm model was built though Endo method, among the 40 rabbits, 8 died or had severe nervous system syndrome, the other 32 were randomly divided into 4 groups:group A, control group, injection of normal saline to the cisterna magna;group B, subarachnoid hemorrhage;group C, injection of human urinary tissue kallikreins;group D, treated with Nimodipine. The behavior scores, neurological scores and cerebral angiography changes were observed.

RESULTSFood intake obviously decreased and neurological deficit were seen in group B, while which were attenuated in group C and group D, and group A was normal. Comparing the diameter of basilar artery was (1.9 +/- 0.3) mm before SAH, the diameter of group B 4 d later was (1.5 +/- 0.3) mm, 7 d later (1.4 +/- 0.3) mm, the difference was significant (P < 0.05). Comparing with group C on the day 4th and 7th, the diameters of basilar artery were significantly different (P < 0.001). Comparing with group D on the day 4th, 7th and 14th, there was no obvious improvement.

CONCLUSIONUrinary kallidinogenase and Nimodipine can obviously alleviate symptomatic cerebral vasospasm in rabbits remarkably, but the former's effect of attenuating vasospasm is better than that of Nimodipine.

Animals ; Disease Models, Animal ; Female ; Humans ; Male ; Nimodipine ; therapeutic use ; Rabbits ; Random Allocation ; Tissue Kallikreins ; therapeutic use ; Vasodilator Agents ; therapeutic use ; Vasospasm, Intracranial ; drug therapy

To prolong serum half-life of human kallikrein (hK) and enhance its secretion rate, we modified hK gene and constructed a new form of recombinant hK protein (hK'-Fc). We amplified hK gene and Fc sequence, replaced the signal peptide of hK gene with murine signal peptide, constructed native expression plasmid of pcDNA-hK-Fc and modified expression plasmid of pcDNA-hK'-Fc, then transfected to CHO cells respectively. After the stable cell lines were screened, we compared the secretion rate between native fusion protein and modified fusion protein, purified fusion protein through Protein A+G affinity chromatography column and investigated the bioactivity of fusion protein. The results showed that recombinant vectors encoding fusion protein hK-Fc and hK'-Fc were constructed successfully; CHO cell lines stably secreting fusion protein were obtained, the yield is higher than 11 mg/L; Secretion rate was enhanced by 5-10 times after the signal peptide of fusion protein was modified; Fusion protein has enzymatic activity in vitro. The above results could promote the following researches on serum half-life of the fusion protein and develop a new stroke medicine with better clinical efficacy.
Animals ; CHO Cells ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Cricetinae ; Cricetulus ; Humans ; Mice ; Protein Sorting Signals ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; physiology ; Tissue Kallikreins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study the expression of the kallikreins-kinins system in the corpus cavernosum of rats.

METHODSThe expression of tissue kallikrein I and kinin B2 receptor gene in the corpus cavernosum and heart of adult rats was detected by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe tissue kallikrein I and kinin B2 receptor were detected in the corpus cavernosum as well as in the heart of the rats and the contents were similar.

CONCLUSIONA kallikreins-kinins system exists in the corpus cavernosum of rats, and the content is rich, almost similar to that in the heart.

Animals ; Male ; Myocardium ; metabolism ; Penis ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptor, Bradykinin B2 ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Kallikreins ; biosynthesis ; genetics

The purpose of this study was to determine whether contemporary active surveillance (AS) protocols could sufficiently discriminate significant from indolent tumors in men with low-risk prostate cancer. We retrospectively analyzed 312 patients with low-risk prostate cancer treated with radical prostatectomy. After exclusion of patients with fewer than 10 cores taken at biopsy and those who received neo-adjuvant treatment, 205 subjects satisfied the final inclusion criteria. Five widely accepted AS protocols were employed in this study. A total of 82.0% of the patients met the inclusion criteria of at least one protocol, and 18% did not meet any criteria of published AS protocols. A significant proportion of patients had non-organ-confined disease (8.6% to 10.6%) or a Gleason score of 7 or greater (18.6% to 23.9%) between the different AS criteria. Among patients who did not meet any AS criteria, 32.4% of patients had a pathologically insignificant cancer. Our results indicated a significant adverse pathology in patients who met the contemporary AS protocols. On the other hand, some patients in whom expectant management would be appropriate did not meet any criteria of published AS protocols. None of the clinical or histological criteria reported to date is able to sufficiently discriminate aggressive tumors from indolent ones.
Aged ; Humans ; Kallikreins/blood ; Male ; Middle Aged ; Neoplasm Grading ; Prostate/*pathology ; Prostate-Specific Antigen/blood ; Prostatectomy ; Prostatic Neoplasms/*pathology/surgery ; Retrospective Studies ; Risk Assessment ; Treatment Outcome ; *Watchful Waiting