Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC₅₀) was calculated by using 1.25×10⁸ CFU/ml,2.50×10⁸ CFU/ml,3.75×10⁸ CFU/ml,5.00×10⁸ CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC₅₀ concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC₅₀=2.5×10⁸ CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.
Animals ; Sheep ; Larva/microbiology* ; Virulence ; Candida glabrata ; Agar ; Moths/microbiology* ; Esterases ; Aspartic Acid Proteases ; Lipase

Cathepsin D (CatD, EC 3.4.23.5) is a member belonging to the subfamily of aspartic endopeptidases, which are classified into the MEROPS clan AA, family A1. Helminth parasites express a large set of different peptidases that play pivotal roles in parasite biology and pathophysiology. However, CatD is less well known than the other classes of peptidases in terms of biochemical properties and biological functions. In this study, we identified 2 novel CatDs (CsCatD1 and CsCatD2) of Clonorchis sinensis and partially characterized their properties. Both CsCatDs represent typical enzymes sharing amino acid residues and motifs that are tightly conserved in the CatD superfamily of proteins. Both CsCatDs showed similar patterns of expression in different developmental stages of C. sinensis, but CsCatD2 was also expressed in metacercariae. CsCatD2 was mainly expressed in the intestines and eggs of C. sinensis. Sera obtained from rats experimentally infected with C. sinensis reacted with recombinant CsCatD2 beginning 2 weeks after infection and the antibody titers were gradually increased by maturation of the parasite. Structural analysis of CsCatD2 revealed a bilobed enzyme structure consisting of 2 antiparallel β-sheet domains packed against each other forming a homodimeric structure. These results suggested a plausible biological role of CsCatD2 in the nutrition and reproduction of parasite and its potential utility as a serodiagnostic antigen in clonorchiasis.
Animals ; Aspartic Acid Endopeptidases ; Biology ; Cathepsin D ; Cathepsins ; Clonorchiasis ; Clonorchis sinensis ; Eggs ; Helminths ; Humans ; Intestines ; Metacercariae ; Ovum ; Parasites ; Peptide Hydrolases ; Rats ; Reproduction

Cysteine and aspartic proteases possess high elastolytic activity and might contribute to the degradation of the abdominal aortic aneurysm (AAA) wall. The aim of this study was to analyze, in detail, the proteases (cathepsins B, D, K, L and S, and inhibitor cystatin C) found in human AAA and healthy aortic tissue samples. The vessel walls from AAA patients (n=36) and nonaneurysmal aortae (n=10) were retrieved using conventional surgical repair and autopsy methods. Serum samples from the same AAA patients and 10 healthy volunteers were also collected. Quantitative expression analyses were performed at the mRNA level using real-time reverse transcriptase-PCR (RT-PCR). Furthermore, analyses at the protein level included western blot and immunoprecipitation analyses. Cellular sources of cysteine/aspartic proteases and cystatin C were identified by immunohistochemistry (IHC). All cysteine/aspartic proteases and cystatin C were detected in the AAA and control samples. Using quantitative RT-PCR, a significant increase in expression was observed for cathepsins B (P=0.021) and L (P=0.018), compared with the controls. Cathepsin B and cystatin C were also detected in the serum of AAA patients. Using IHC, smooth muscle cells (SMCs) and macrophages were positive for all of the tested cathepsins, as well as cystatin C; in addition, the lymphocytes were mainly positive for cathepsin B, followed by cathepsins D and S. All cysteine/aspartic proteases analyzed in our study were detected in the AAA and healthy aorta. The highest expression was found in macrophages and SMCs. Consequently, cysteine/aspartic proteases might play a substantial role in AAA.
Aged ; Aorta/enzymology ; Aortic Aneurysm, Abdominal/*enzymology ; Aspartic Acid Proteases/genetics/*metabolism ; Case-Control Studies ; Cathepsins/genetics/metabolism ; Cysteine Proteases/genetics/*metabolism ; Humans ; Lymphocytes/enzymology ; Macrophages/enzymology ; Middle Aged ; Myocytes, Smooth Muscle/enzymology ; RNA, Messenger/genetics/metabolism

There is considerable need for reliable prognostic markers to guide clinicians in management decisions for the breast cancer. The cell cycle is governed by a family of cyclin-dependent kinases(Cdks), regulated by associated cyclin p27, a cyclin dependent kinase inhibitors, regulates progression from GI into S phase by inhibiting cyclin/cdks complex. This study was performed to evaluate the association between p27 expression, determined by immunohistochemical stain, and various histopathologic features in breast cancer. It was also determined whether p27 expression had the significance as the prognostic factorin the breast cancer patients. 45 patients who got the relatively good preserved paraffin blocks among the 100 patients was chosen for immunohistochemical staining against p27, cyclin E, c-erbB2, cathepsin D and p53 and reexamined their unclear and histological grades from Jan. 1989 through Dec. 1992. As a results, the patients with negative expression of p27 had more metastatic axillary nodes than those with positive expression. (5.87+/-1.87 vs 1.14+/-0.54, p=0.021) p27 negative group got worse nuclear grade than p27 negative group but beyond statistical significance. (48.4% vs 28.6%, p=0.281) On univariate analysis, primary tumor size, status of axillary nodes and the expression of p27 were the significant prognostic factors affecting overall survival rates. In particular, p27 positive group had better outcomes on 5 years survival rate than p27 negative group. (92.31+/-7.39% vs 73.33+/-8.07%, p=0.0441) but didn't affect the disease free survival with statistical significance. On multivariate analysis, the primary tumor size and axillary node were significant prognostic factors on overall and disease free survival. In conclusion, despite relativeiy small size of this study group, considering that p27 negative group got the more metastatic node and worse overall survival, 27 expression might be a novel prognostic indicator in the breast cancer.
Breast Neoplasms* ; Breast* ; Cathepsin D ; Cell Cycle ; Cyclin E* ; Cyclins* ; Disease-Free Survival ; Humans* ; Multivariate Analysis ; Paraffin ; Phosphotransferases ; S Phase ; Survival Rate

Female phenotype of a 46,XY male may originates from male pseudohermaphroditism due to 17alpha-hydroxylase deficiency. Lack of cortisol increases adrenocorticotropic hormone (ACTH) and mineralocorticoid production, leading to low renin hypertention and hypokalemia. A 41-year-old phenotypic female presented primary amenorrhea and hypertension. In the hormonal profile, the levels of serum estradiol, testosterone, rennin, and cortisol were decreased and ACTH and deoxycorticosterone were increased. Laparoscopic bilateral gonadectomy was performed, and corticosteroid, antihypertensive drugs, and estrogen were administered. We report this case with a brief review of the literatures.
46, XY Disorders of Sex Development* ; Adrenal Hyperplasia, Congenital ; Adrenocorticotropic Hormone ; Adult ; Amenorrhea ; Antihypertensive Agents ; Chymosin ; Desoxycorticosterone ; Estradiol ; Estrogens ; Female ; Humans ; Hydrocortisone ; Hypertension ; Hypokalemia ; Male* ; Phenotype ; Renin ; Testosterone

The renal manifestations of systemic sclerosis include proteinuria, hypertension, azotemia, and renal crisis. Two types of scleroderma renal crisis (SRC) are recognized. Typical SRC is a syndrome consisting of acute-onset malignant hypertension accompanied by rapidly progressive renal failure, hypertensive retinopathy, and elevated plasma renin activity. The other type is normotensive renal failure, which is generally accompanied by antineutrophil cytoplasmic autoantibody (ANCA)-positive crescentic glomerulonephritis. A 51-year-old woman with scleroderma without marked dermatological change developed ANCA-related renal failure. She had neither malignant hypertension nor an elevated plasma rennin concentration. Renal biopsy showed crescentic glomerulonephritis (pauci-immune type), and the myeloperoxidase-specific ANCA (MPO-ANCA) titer was elevated at 1015 AAU. She was cured using steroid pulse therapy, combined with an angiotensin-converting-enzyme inhibitor and angiotensin-II receptor blocker
Antibodies, Antineutrophil Cytoplasmic ; Azotemia ; Biopsy ; Chymosin ; Cytoplasm ; Female ; Glomerulonephritis ; Humans ; Hypertension ; Hypertension, Malignant ; Hypertensive Retinopathy ; Isonipecotic Acids ; Middle Aged ; Plasma ; Proteinuria ; Renal Insufficiency ; Renin ; Scleroderma, Systemic

Primary aldosteronism is a disease that the stimulus for the excessive aldosterone production residues within the adrenal gland. It was first described by conn in 1955. And many cases were reported by physicians at present in the world. But it is relatively rare in Korea, probably due to lack of attention and medical facilities. Only about 13 cases have been reported at present. The clinical, biochemical feature in l case of primary aldosteronism caused by adrenal hyperplasia that was diagnosed at Yeungnam University Hospital was observed and the following result were obtained. 1. Clinical feature: The present case was 27-year-old woman who was admitted due to general weakness and easy fatigability. The above mentioned chief complaints occurred 8 months prior to admission when she delivered of second baby by cesarean section. Symptoms such as above chief complaints, intermittent muscle paralysis and cramping were noticed. Trousseau's sign was also present. The average blood pressure ranged from 170/90 to 200/120 2. Biochemical abnormalities: Severe hypokalemia lower than 2.5 mEq/L was presented and 24 hours urine potassium showed markedly increased urinary loss (228 mEq/day). Plasma rennin activity was decreased under normal range with furosemide administration. (Basal renin; 0.01 ng/ml/hr, stimulated rennin 0.12 ng/ml/hr). Saline suppression test revealed markedly elevated levels of aldosterone higher than normal range (Basal aldosterone; 320.68 pg/ml stimulated aldosterone; 451.86 pg/ml). And posture test showed decreased plasma rennin activity and increased plasma aldosterone level. 3. Adrenal CT scan revealed no abnormal findings. 4. Treatment and course: Spironolactione was given at OPD with regular follow-up. Her blood pressure ranged from 150/90 to 160/100 and symptoms were improved. The effect of treatment was satisfactory and further follow up would be performed.
Adrenal Glands ; Adult ; Aldosterone ; Blood Pressure ; Cesarean Section ; Chymosin ; Female ; Follow-Up Studies ; Furosemide ; Humans ; Hyperaldosteronism* ; Hyperplasia* ; Hypokalemia ; Korea ; Muscle Cramp ; Paralysis ; Plasma ; Posture ; Potassium ; Pregnancy ; Reference Values ; Renin ; Tomography, X-Ray Computed

PURPOSE: The purpose of this study was to find out what clinicopathologic or immunohistochemical parameter that may affect FDG uptake of primary tumor in PET/CT scan of the gastric carcinoma patient. MATERIALS AND METHODS: Eighty-nine patients with stomach cancer who underwent pre-operative FDG PET/CT scans were included. In cases with perceptible FDG uptake in primary tumor, the maximum standardized uptake value (SUVmax) was calculated. The clinicopathologic results such as depth of invasion (T stage), tumor size, lymph node metastasis, tumor differentiation and Lauren's classification and immunohistochemical markers such as Ki-67 index, expression of p53, EGFR, Cathepsin D, c-erb-B2 and COX-2 were reviewed. RESULTS: Nineteen out of 89 gastric carcinomas showed imperceptible FDG uptake on PET/CT images. In cases with perceptible FDG uptake in primary tumor, SUVmax was significantly higher in T2, T3 and T4 tumors than T1 tumors (5.8+/-3.1 vs. 3.7+/-2.1, p=0.002). SUVmax of large tumors (above or equal to 3 cm) was also significantly higher than SUVmax of small ones (less than 3 cm) (5.7+/-3.2 vs. 3.7+/-2.0, p=0.002). The intestinal types of gastric carcinomas according to Lauren showed higher FDG uptake compared to the non-intestinal types (5.4+/-2.8 vs. 3.7+/-1.3, p=0.003). SUVmax between p53 positive group and negative group was significantly different (6.0+/-2.8 vs. 4.4+/-3.0, p=0.035). No significant difference was found in presence of LN metastasis, tumor differentiation, Ki-67 index, and expression of EGFR, Cathepsin D, c-erb-B2 and COX-2. CONCLUSION: T stage of gastric carcinoma influenced the detectability of gastric cancer on FDG PET/CT scan. When gastric carcinoma was perceptible on PET/CT scan, T stage, size of primary tumor, Lauren's classification and p53 expression were related to degree of FDG uptake in primary tumor.
Cathepsin D ; Humans ; Lymph Nodes ; Neoplasm Metastasis ; Stomach Neoplasms

PURPOSE: To evaluate the prognostic significances of p53 and cathepsin D in the prostatic carcinoma, we compared them to other prognostic factors, such as nuclear grade and clinical stage. MATERIALS AND METHODS: The material consisted of 40 paraffin-embedded, primary prostate carcinomas. We examined the expression of p53 and cathepsin D using immunohistochemical staining and compared their expression with the grade and stage. RESULTS: The expressions of p53 were noted in the nucleus of tumor cells and cathepsin D were noted in the cytoplasm of tumor cells. Thirteen of 40 tumors were positive for p53. There were more expressing p53 in samples (40%) from prostatic cancer with a high Gleason score group than in samples (28%) from prostatic cancer with low Gleason score group. The expression of p53 was 22% in clinical stage B and C groups and 35% in clinical stage D group. These results showed that p53 expression was not statistically correlated with Gleason score and clinical stage, but there were trends to increased p53 expression with high Gleason score and progressed clinical stage (p>0.05). Progressed clinical stage group showed higher expression of cathepsin D than early clinical stage group. However, there were no statistical correlations between expression of cathepsin D and Gleason score, and clinical stage (p>0.05). CONCLUSION: These results suggest that the overexpression of p53 and cathepsin D may be associated with tumor differentiation and clinical stage, but have limited prognostic value in prostatic carcinoma.
Cathepsin D* ; Cathepsins* ; Cytoplasm ; Neoplasm Grading ; Prostate ; Prostatic Neoplasms

Twenty six cases of primary adenocarcinoma of the prostate, ranging from 4 to 9 according to Gleason's summing score, were studied. Immunoreactivity was evaluated using the rabbit polyclonal anti-Cathepsin D antibody (CD), a mouse monoclonal MMP-2 antibody (MMP-2), and a tissue inhibitor metalloproteinase (TIMP) in formalin-fixed, paraffin-embedded prostatic tissue. Immunohistochemical staining was scored by summing the intensity of staining (0 to 3+) weighted by the percentage of tumor staining at each intensity (H score, theoretical range 0 to 300). For CD, the tumor cells showed diffuse cytoplasmic immunoreactivity in all 26 cases (100%). For MMP-2 the tumor cells showed cytoplasmic immunoreactivity in 17 of 26 cases (65.38%). As the Gleason grade increased the expression of CD increased (P=0.0027). The reactivity of CD was significantly correlated with the Gleason's score (R=0.65637), but, the reactivity of MMP-2 was not correlated. There were no significant correlations between each of the CD and the MMP-2 scores, and stage. TIMP expression was predominantly localized in the stroma rather than in the cancer cells themselves. We believe that 1) CD and MMP-2, both immunohistochemically detectable in a majority of prostate adenocarcinoma, may play a role in determination of the invasive or metastatic property, 2) the enhanced TIMP expression in the stroma may be associated with the response to cancer invasion.
Adenocarcinoma ; Animals ; Cathepsin D* ; Cathepsins* ; Cytoplasm ; Mice ; Prostate*