[Analysis of a family with 11ß-hydroxylase deficiency due to a mutation in the CYP11B1 gene].

This study reports a family of patients with 11ß-hydroxylase deficiency (11ß-OHD) caused by a novel mutation in the CYP11B1 gene, and analyzes its clinical and genetic characteristics. The clinical data of a patient with intractable hypertension at Air Force Medical Center on May 16, 2014 were retrospectively analyzed. The patient was clinically diagnosed with congenital adrenal cortical hyperplasia. The clinical data of the patient were further collected and the peripheral blood samples of the patient, his parents and his sister were collected for CYP11B1(NM_000497) gene sequencing, suggesting that the patient had compound heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9 and exon 5:c.905_907 delATGinsTT, p.Asp302Valfs*23, both of which were pathogenic variants. The patient's father and sister carried heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9, and the mother carried heterozygous mutations in exon 5:c.905_907delATGinsTT, p.Asp302Valfs*23. This study is the first to report a new compound heterozygous mutation in exon 1:c.199delG and exon 5 c.905_907 delATGinsTT of CYP11B1 gene, enriching the database of 11ß-OHD mutations and providing information to further understand the genetic mechanism of the disease.

Correlation between neuregulin receptor degradation protein-1 and Crohn's disease.

Neuregulin receptor degradation protein-1 (Nrdp1) is a newly discovered E3 ligase that plays a role in the apoptosis process of multiple diseases. Previous studies has shown that Nrdp1 exerted a proapoptotic effect in cardiac diseases. The purpose of this study is to investigate the potential involvement of Nrdp1 in the pathological processes of inflammatory bowel disease (IBD). To create a mouse model of experimental colitis, trinitrobenzenesulfonic acid (TNBS) was administered and the severity of colitis was assessed based on changes in weight and histological scores. Using Western blot and immunohistochemistry, significant increase in Nrdp1 expression was observed in intestinal epithelial cells (IECs). This was accompanied with the up-regulation of cleaved PARP and active caspase-3 in IECs, indicating a potential function in IECs. To study this further, we built an in vitro model of tumor necrosis factor-alpha (TNF-α)-induced apoptosis using human IEC line HT-29 cells. When Nrdp1 was knocked down, a decrease in apoptosis was observed, suggesting that Nrdp1 may play a proapoptotic role in IEC apoptosis. The mechanism behind this phenomenon is associated with the suppression of downstream targets of Nrdp1, such as protein kinase B (AKT). Furthermore, immunohistochemistry analysis in patients with Crohn's disease (CD) and normal controls supported the same results as observed in experimental colitis. We conclude that Nrdp1 may be a promising new therapeutic target for ameliorating IBD in humans.

The progression of pancreatic cancer cells is promoted by a long non-coding RNA LUCAT1 by activating AKT phosphorylation.

OBJECTIVE: In many cancers, long non-coding RNAs (lncRNA) are largely involved; they can regulate cell proliferation, migration, and invasion. However, the research of lncRNA regulation on pancreatic ductal adenocarcinoma is vacant. The aim of this article was to lucubrate the specific role of lncRNA LUCAT1 in regulating the progression of pancreatic cancer. PATIENTS AND METHODS: Pancreatic cancer and adjacent tissues were collected, and the expression of LUCAT1, one potential involved LucRNA, was measured using real-time qPCR (RT-qPCR). Different pathological types of pancreatic cancer cell lines were cultured, and the expression difference of LncRNA LUCAT1 was detected by RT-qPCR, and two cell lines were selected for downstream experiments. si-RNA was used to knockdown the expression of LUCAT1, comparing the difference in expression of LUCAT1, characterizing cell proliferation by MTT and BrdU staining, detecting apoptosis, and cell cycle changes by flow cytometry. Meanwhile, Western blotting was used for the detection of cyclin expression and thus investigate two important associated signaling pathways. Besides, the expression of signaling pathway was validated by signaling inhibitor. RESULTS: In comparison to normal cells, LUCAT1 was highly expressed in human pancreatic cancer cell lines (p<0.05). The higher expression of LUCAT1 resulted in enhanced pathogenesis of PDA cells and motivated the development to S phase by regulation of cyclin D1, CDK4. Furthermore, LUCAT1 promoted PDA cells development by inducing AKT's and p38 MAPK's phosphorylation. CONCLUSIONS: LUCAT1, as the key factor, played a positive role in the proliferation and invasion of pancreatic cells via AKT/MAPK signaling.

Effects of habitat fragmentation on the genetic diversity and differentiation of Dendrolimus punctatus (Lepidoptera: Lasiocampidae) in Thousand Island Lake, China, based on mitochondrial COI gene sequences.

Thousand Island Lake (TIL) is a typical fragmented landscape and an ideal model to study ecological effects of fragmentation. Partial fragments of the mitochondrial cytochrome oxidase subunit I gene of 23 island populations of Dendrolimus punctatus in TIL were sequenced, 141 haplotypes being identified. The number of haplotypes increased significantly with the increase in island area and shape index, whereas no significant correlation was detected between three island attributes (area, shape and isolation) and haplotype diversity. However, the correlation with number of haplotypes was no longer significant when the 'outlier' island JSD (the largest island) was not included. Additionally, we found no significant relationship between geographic distance and genetic distance. Geographic isolation did not obstruct the gene flow among D. punctatus populations, which might be because of the high dispersal capacity of this pine moth. Fragmentation resulted in the conversion of large and continuous habitats into isolated, small and insular patches, which was the primary effect on the genetic diversity of D. punctatus in TIL. The conclusion to emphasize from our research is that habitat fragmentation reduced the biological genetic diversity to some extent, further demonstrating the importance of habitat continuity in biodiversity protection.

Value of microsurgical varicocelectomy for severe oligo-asthenospermia patients failed in fertilization assisted by in vitro fertilization.

OBJECTIVE: To investigate the clinical effect of microsurgical varicocelectomy on severe oligo-asthenospermia patients failing in fertilization assisted by intracytoplasmic sperm injection. PATIENTS AND METHODS: From January 2013 to August 2014, forty-nine patients with severe oligo-asthenospermia and serious varicoceles were treated by microsurgical varicocelectomy after failing in fertilization assisted by intracytoplasmic sperm injection (ICSI), eleven of whom had varicoceles on the left side and thirty-eight had bilateral varicoceles. Patients were followed up for the natural pregnancy condition, changes of routine semen parameters and reproductive hormone level and the embryonic development and outcome of next IVF-ET (ICSI) cycles within 6 months. RESULTS: After surgery, 61.2% (30/49) of spouses obtained clinical pregnancy. Among whom 22.4% (11/49) were naturally pregnant, 32.65% (16/49) were conceived after second IVF-ET assistance, and 6.1% (3/49) were conceived with the third or further assistance of ICSI-ET. The overall miscarriage rate was 16.7% (5/30). All of the patients had improvement in the sperm concentration and forward motility. The sperm concentration increased from (10.53 ± 8.76) × 106/ml to (20.23 ± 11.76) × 106/ml. The ratio of forward motile sperm was increased to (30.52 ± 18.78) % from (8.75.52 ± 6.36) % (p < 0.01). The serum total testosterone (T) improved from (2.19 ± 1.03) ng/ml to (4.05 ± 0.64) ng/ml (p < 0.05). Serum follicle-stimulating hormone (FSH) changed from (5.23 ± 1.26) mIU/ml to (3.76 ± 2.22) mIU/ml after the procedure. Luteinizing hormone (LH) changed from (4.38 ± 1.36) to (3.98 ± 1.38) mIU/ml. Estrogen (E2) changed from 40.28 ± 7.26 pg/ml to 35.24 ± 5.75 pg/ml. Prolactin (PRL) level elevated from (18.24 ± 4.28) to (17.16 ± 2.16) ng/ml (p > 0.05). The fertility rate of in vitro fertilization significantly improved to (83.36 ± 19.36) % from (72.36 ± 17.88) % (p < 0.05). The rate of 2PN ratio increased from (66.73 ± 17.93) % to (75.96 ± 20.39) %. The cleavage rate increased from (83.26 ± 32.33) % to (90.35 ± 23.66). The abnormal fertility rate were (5.36 ± 12.58) % and (7.26 ± 13.89) % before and after the procedure (p > 0.05), while the rate of high-quality embryos increased significantly from (34.36 ± 33.27) % to (55.67 ± 23.36) % (p < 0.05). The rate of transferable embryos remained without significant change (70.67 ± 30.6% before and 60.53 ± 30.27% after the procedure). The anabiosis rate of frozen embryo increased from (66.32 ± 30.69) % to (89.72 ± 29.69) %. The further blastocyst rate improved from (10.98 ± 9.7) % to (30.27 ± 15.33) % (p < 0.01). CONCLUSIONS: The microsurgical varicocelectomy effectively improved sperm parameters, the fertility rate of oocyte fertilized in vitro and the anabiosis rate and blastocyst rate of the frozen embryo for on patients with severe oligo-asthenospermic, and further increased the odds of natural pregnancy, the rate of high-quality embryos and the success rate of in vitro fertilization.

Mortality rate for children under 5 years of age in China from 1996 to 2006.

OBJECTIVES: To study the change in mortality rate for children under 5 years of age in China over the past decade, and to evaluate China's progress in achieving Millennium Development Goal 4. STUDY DESIGN: Population-based descriptive study. METHODS: A population-based survey was conducted through a nationwide multi-level surveillance network. The mortality rate and the leading causes of death for children under 5 years of age were analysed. RESULTS: The mortality rate for children under 5 years of age in China dropped by 54.2% between 1996 and 2006 (from 45.0 per 1000 livebirths to 20.6). During this period, deaths due to pneumonia and diarrhoea dropped by 69.4% and 69.7%, respectively. The proportion of deaths due to pneumonia dropped from 23.4% in 1996 to 15.6% in 2006, and the proportion of deaths due to diarrhoea dropped from 5.6% in 1996 to 3.7% in 2006. CONCLUSION: The mortality rate for children under 5 years of age in China dropped remarkably from 1996 to 2006. This reduction was mainly due to a significant decrease in deaths due to pneumonia and diarrhoea. Based on the survey results, China should be able to achieve Millennium Development Goal 4.

Clinical observation of 73 nasopharyngeal carcinoma patients treated by helical tomotherapy: the China experience.

The preliminary short-term clinical outcome of 73 nasopharyngeal carcinoma (NPC) patients treated with helical tomotherapy at our cancer institute has been evaluated. Between September 2007 and September 2009, 73 newly diagnosed NPC patients were treated with helical tomotherapy. The distributions of clinical stages according to the UICC 2002 Staging System were: 6, 27, 24, and 16 for Stage I, IIa-b, III, and IVa-b, respectively. The prescription dose was 70-74 Gy/33F to planning gross tumor volume containing the primary tumor and positive lymph nodes, with 60-62.7 Gy/33F to high risk planning target volume, while delivering 52-56 Gy/33F to low risk planning target volume. Twenty-four patients were treated with radiation therapy as single modality, 25 with concurrent cisplatin-based chemotherapy with or without anti-EGFR monoclonal antibody therapy, and 24 with concurrent anti-EGFR monoclonal antibody therapy. Setup errors were analyzed. Side-effects were evaluated with the established RTOG/EORTC criteria. Average beam-on-time was 468.8 sec/F (396.7-696.1 sec). The setup errors in the lateral, longitudinal and vertical directions were 0.00 ± 1.79 mm, -0.55± 2.17 mm and 0.38 ± 1.43 mm, corresponding to 3.80 mm, 4.20 mm, and 2.46 mm as the CTV-PTV margin in these directions. The grade 0, 1, 2 and 3 acute skin toxicity was 2.7%, 76.7%, 13.8% and 6.8%; the grade 0, 1, 2 and 3 acute mucositis was 1.4%, 32.9%, 60.2% and 5.5%; and the grade 0, 1, 2 and 3 acute xerostomia was 4.0%, 45.3%, 50.7% and 0, respectively. Only 5 patients suffered from grade 3 or 4 leucopenia. Xerostomia resolved with passing of time and no grade 2 or more xerostomia was noted one year after radiation therapy. Concurrent chemotherapy significantly increased incidence of severe acute toxicities. One month after radiation therapy the remission rates of primary tumor and positive lymph nodes were 91.8% and 98.1%, respectively. The median follow-up was 14.8 months. The one-year relapse-free survival, distant metastasis-free survival and overall survival was 95.6%, 97.2% and 94.8%, respectively. In conclusion, the incidence of severe acute toxicities and late xerostomia was relatively infrequent for NPC patients treated with helical tomotherapy. The long-term clinical outcome for these patients is under investigation.

Clinical dosimetric study of three radiotherapy techniques for postoperative breast cancer: Helical Tomotherapy, IMRT, and 3D-CRT.

This paper is to investigate the dosimetric characteristics of Helical Tomotherapy (HT), step-and-shoot intensity-modulated radiation therapy (SaS-IMRT) and three-dimensional conformal radiation therapy (3D-CRT) for the postoperative breast cancer as well as their dosimetric comparison of the normal tissues. CT images of 10 postoperative patients with early stage breast cancer were transferred into HT, SaS-IMRT and 3D-CRT planning systems respectively after the target region and normal tissues were outlined by the same physician to assure the contour consistency. Each prescribed dose for three different modalities of plans was given to a total of 50 Gy in 25 fractions. Doses and irradiated volumes in heart, lungs, as well as conformity index (CI) and homogeneity index (HI) were evaluated for detailed comparison. All three plans showed appropriate coverage for the prescribed target dose in the dosimetric comparison. The CI in HT and SaS-IMRT as well as 3D-CRT was 0.68 ± 0.12, 0.58 ± 0.08 and 0.40 ± 0.08, respectively. The HI were 1.10 ± 0.03, 1.14 ± 0.02 and 1.17 ± 0.04, which appeared intergroup significant differences (p < 0.05). V5, V10, as well as V20 of the heart were smallest in 3D-CRT than HT and SaS-IMRT. V5 of the ipsilateral lung was the smallest in 3D-CRT than HT and SaS-IMRT (p < 0.05); However, V20 and V30 were smaller in HT and SaS-IMRT than 3D-CRT (p < 0.05). V5 of the contralateral lung was the smallest in 3D-CRT than other groups, with V10~V30 were basically similar in numeric values with not obvious discrepancy. Comparing with SaS-IMRT and 3D-CRT, HT technique in treating breast cancer had the best conformity and homogeneity index as well as steepest dose gradient due to its highly modulated beamlets with rotational technique. The heart volume irradiated was the smallest in conventional 3D-CRT, with SaS-IMRT was the largest among the three techniques, as expected. The volume of the contralateral lung irradiated was the smallest in 3D-CRT than other groups. V5 of the ipsilateral lung was the smallest in 3D-CRT than other two groups. V10~V30 in HT and SaS-IMRT were similar and better than 3D-CRT dosimetrically. We conclude that HT technique had advantages over SaS-IMRT and 3D-CRT based on the dosimetric comparison in this study, especially in the high dose region of ipsilateral lung, target homogeneity and dose uniformity.

Suppression of 5-lipoxygenase gene is involved in triptolide-induced apoptosis in pancreatic tumor cell lines.

Pancreatic adenocarcinoma is characterized by a poor prognosis and lack of response to conventional therapy. The purpose of this study was to investigate the effects of triptolide (TL) on proliferation and apoptosis of pancreatic cancer cells in vitro. We found that TL induced prominent growth inhibition and apoptosis in human pancreatic cell lines. In addition, TL treatment significantly down-regulated 5-lipoxygenase (5-LOX) expression, as well as downstream leukotriene B4 (LTB4) production, in these cell lines. Furthermore, overexpression of 5-LOX in SW1990 cell lines or exogenous LTB4 made them more resistant to TL-induced apoptosis, which was correlated with increased Bcl-2 expression. Taken together, these findings suggest that inhibition of the 5-LOX pathway of arachidonic acid metabolism is associated with the anti-proliferation activity of TL. We also provide evidence that TL has clinical therapeutic value for patients with pancreatic cancer.

New annonaceous acetogenins from the roots of Uvaria calamistrata.

Five new annonaceous acetogenins, calamistrins C-G (1-5), were isolated from an ethanolic extract of the roots of Uvaria calamistrata. Compounds 1-3 were mono-THF ring acetogenins; compounds 4 and 5 were bis-THF acetogenins, with the THF rings from C-18 to C-25. The absolute configurations of 3, 4, and 5 as well as the partial absolute configurations of 1 and 2 were determined by (13)C NMR spectroscopy and advanced Mosher methodology.

Polyoxygenated bipyridine, pyrrolylpyridine, and bipyrrole alkaloids from Speranskia tuberculata.

Five novel polyoxygenated alkaloids, speranculatines A-C (3-5), speranskilatine A (6), and speranberculatine A (7), have been isolated from Speranskia tuberculata. Compounds 3-5, 6, and 7, have bipyridine, pyrrolylpyridine, and bipyrrole skeletons, respectively. This is the first time that these three alkaloid structural types have been reported. The structures of 3-7 were elucidated by spectroscopic methods, including 2D NMR techniques and X-ray crystallographic analysis.

Calamistrins A and B, two new cytotoxic monotetrahydrofuran annonaceous acetogenins from Uvaria calamistrata.

Two new bioactive monotetrahydrofuran acetogenins, calamistrins A (1) and B (2), and two known compounds, uvarigrin (3) and uvarigranin (4), have been isolated from the roots of Uvaria calamistrata. The structures of the new compounds were elucidated by spectroscopic and chemical methods. The absolute stereochemistry of the stereogenic centers was established by Mosher ester methodology.

Styryllactones from the rhizomes of Goniothalamus griffithii.

Three new styryl-lactones 8-acetylgoniofufurone(1), 7-acetylgonio-pypyrone(3), and 5-acetylgoniopypyrone(4), along with ten known compounds, goniofufurone(2), goniopypyrone(5), goniothalamin, goniothalenol, (+)-isoaltholactone, goniodiol, 7-acetylgoniodiol, goniotriol, 8-acetylgoniotriol, 9-deoxygoniopypyrone were isolated from the rhizomes of Goniothalamus griffithii Hook f. et. Thoms. Their structures were elucidated by IR, MS, NMR spectra and chemical evidence. All compounds showed cytotoxic activities against human cancer cell lines.

New polyoxygenated cyclohexenes from Uvaria calamistrata.

Five new polyoxygenated cyclohexenes, named uvacalol A (1), B (2), C (3), D (4) and E (5) were isolated from the roots of Uvaria calamistrata. On the basis of spectral analysis and chemical derivatization, including the preparation of Mosher esters, the structures of compound 1-5 were established as (2R,3S,4R,5S)-2-acetoxyl-5-ethoxyl-1-benzoyloxymethylcyclohex-1(6)-ene3,4-diol-3-benzoate, (2R,3S,4R,5S)-2-acetoxyl-5-ethoxyl-1-benzoyloxymethylcyclohex-1(6)-ene-3,4-diol-4-benzoate, (2R,3S,4R,5S)-5-ethoxyl-1-benzoyloxymethylcyclohex-1(6)-ene-2,3,4-triol-3-benzoate, (2R,3S,4R,5S)-3-methoxyl-1-benzoyloxymethylcyclohex-1(6)-ene-2,3,5-triol and (2R,3S,4R,5S)-2-acetoxyl-1-benzoyloxymethylcyclohex-1(6)-ene-3,4,5-triol-5-benzoate, respectively.

Antibodies to endometrial transferrin and alpha 2-Heremans Schmidt (HS) glycoprotein in patients with endometriosis.

PROBLEM: Identifying the endometrial antigens inciting autoimmunity is important in setting up an antibody assay for a non-invasive diagnosis and clinical monitoring of endometriosis. METHODS: Two-dimensional gel electrophoresis of endometrial extracts, Western blot analysis, passive hemagglutination and enzyme-linked immunosorbent assay (ELISA), amino acid sequencing and molecular studies were done on chosen antigens. Forty-six women with endometriosis, 4 women with uterine leiomyomata, 4 with pelvic adhesions, 3 with repeat Cesarean sections (conditions that coexist with or predispose to endometriosis) and 46 controls participated. RESULTS: Antigens with molecular weights (MW) of 64 kDa [isoelectric point (pI) of 3.5-4.0] and 72 kDa (pI of 4.5) bound to IgG in all patients with endometriosis, but not the controls. Amino acid sequencing of the proteins revealed that they had homology to alpha 2-Heremans Schmidt (HS) glycoprotein (MW: 64 kDa) and transferrin (MW: 72 kDa). Endometriosis patients had significant antibody levels to these two proteins (predictive value of 80-90%). The analysis of patients' endometrial RNA detected the message for alpha 2-HS glycoprotein and transferrin. Albumin (pI 5.5) and collagen (pI 3.5) failed to elicit antibody responses. CONCLUSIONS: Patients with endometriosis have significant antibodies to endometrial transferrin and alpha 2-HS-glycoprotein. We can effectively use an antibody assay using these antigens for diagnosing endometriosis.

Toxicity of human recombinant interferon-gamma in rats and dogs.

AIM: To study the acute and chronic toxicities of human recombinant interferon-gamma (Hu-rIFN-gamma) in mice, rats, and dogs. METHOD: Twenty mice were administrated Hu-rIFN-gamma (i.m. or i.v.) 4.4 x 10(9) IU m-2 to observe the acute toxicity. In chronic studies, 1 x 10(7), 5 x 10(7), 1 x 10(8) IU m-2 d-1 were given to 80 rats and 5 x 10(5), 5 x 10(7) IU m-2 d-1 were injected to 14 dogs i.m. for 3 months, treatment-related changes were measured in the hematologic, chemical, urinalysis values, ECG and pathologic profile of organs and tissues. RESULTS: The maximal tolerance dose (MTD) i.m. or i.v. in mice was 4.4 x 10(9) IU m-2, 4400 times the recommended clinical dosage (1 x 10(6) IU m-2). No adverse effects were found in chronic toxicity studies. CONCLUSION: Human recombinant interferon-gamma did not produce toxic reaction in rats and dogs.

Sequential activation of two mitogen activated protein (MAP) kinase isoforms in rat skeletal muscle following insulin injection.

Skeletal muscle is a major target of insulin action. The possible role of MAP kinase activation in insulin receptor signaling in muscle was examined. After a 48-hr fast, rats were injected intravenously with insulin or saline, muscles were excised after 3-20 min, homogenized, and MAP kinases were partially purified by ammonium sulfate precipitation and Mono Q chromatography. Activity was assayed as 32P-incorporation into myelin basic protein. Two activity peaks were identified; peak I eluted with approximately 0.1 M NaCl and peak II with approximately 0.2 M NaCl. Three min after insulin injection the activity of peak II increased > 2-fold, peak I was unchanged. After 10 min, the activity of peak II returned toward baseline, while peak I was activated approximately 3-fold. Immunoblots confirmed the presence of MAP kinases eluting with activity peaks I and II; the former as a approximately 41 kDa protein and the latter as a doublet of approximately 42 and approximately 44 kDa. The data suggest sequential activation of two MAP kinases in muscles; the isoform which activates/deactivates rapidly may represent ERK-1, while the more slowly responding isoform may be ERK-2.

Kallistatin: a novel human tissue kallikrein inhibitor. Purification, characterization, and reactive center sequence.

A novel human tissue kallikrein inhibitor designated as kallistatin has been purified from plasma to apparent homogeneity by polyethylene glycol fractionation and successive chromatography on heparin-Agarose, DEAE-Sepharose, hydroxylapatite, and phenyl-Superose columns. A purification factor of 4350 was achieved with a yield of approximately 1.35 mg per liter of plasma. The purified inhibitor migrates as a single band with an apparent molecular mass of 58 kDa when analyzed on SDS-polyacrylamide gel electrophoresis under reducing conditions. It is an acidic protein with pI values ranging from 4.6 to 5.2. No immunological cross-reactivity was found by Western blot analyses between kallistatin and other serpins. Kallistatin inhibits human tissue kallikrein's activity toward kininogen and tripeptide substrates. The second-order reaction rate constant (ka) was determined to be 2.6 x 10(4) M-1 s-1 using Pro-Phe-Arg-MCA. The inhibition is accompanied by formation of an equimolar, heat- and SDS-stable complex between tissue kallikrein and kallistatin, and by generation of a small carboxyl-terminal fragment from the inhibitor due to cleavage at the reactive site by tissue kallikrein. Heparin blocks kallistatin's complex formation with tissue kallikrein and abolishes its inhibitory effect on tissue kallikrein's activity. The amino-terminal residue of kallistatin is blocked. Sequence analysis of the carboxyl-terminal fragment generated from kallistatin reveals the reactive center sequence from P1' to P15', which shares sequence similarity with, but is different from known serpins including protein C inhibitor, alpha 1-antitrypsin, and alpha 1-antichymotrypsin. The results show that kallistatin is a new member of the serpin superfamily that inhibits human tissue kallikrein.