Distribution of yeast isolates from invasive infections and their in vitro susceptibility to antifungal agents: evidence from 299 cases in a 3-year (2010 to 2012) surveillance study.

Invasive yeast infections cause significant morbidity and mortality. Surveillance for the infection is necessary to detect trends in species distribution and antifungal resistance. We performed this retrospective study of yeast infection at Jinling Hospital, Nanjing in China, from year of 2010 to 2012. A total of 341 yeast isolates were obtained from patients with invasive infections in the period. Among these isolates, Candida spp. comprised of the highest percentage of yeast strains (91.8 %), followed by Cryptococcus neoformans (5.9 %) and other non-Candida yeast strains (2.3 %). Bloodstream isolates made up 41.3 % of yeast strains and the isolates from CVC made up 17.3 %. Among Candida spp., C. albicans was the most common species identified from non-blood clinical specimens (42.9 %), but appeared in only 20.8 % of blood isolates (P < 0.001). C. tropicalis was the most prevalent Candida species in the blood samples (28.5 %). Candida spp. was mainly isolated from specimens of the ICU patients, while C. neoformans was mainly isolated from specimens in medical wards. Resistance to FLC occurred in 3.7 % of C. albicans, 9.9 % of C. tropicalis, 74.0 % of C. glabrata, and 4.4 % of C. parapsilosis. Most (>92 %) isolates of C. albicans, C. tropicalis, C. parapsilosis, and C. neoformans strains were susceptible to VRC; However, 26.7 % of isolates of C. glabrata were VRC resistant.

Diagnostic value of immunoglobulin G antibodies against Candida enolase and fructose-bisphosphate aldolase for candidemia.

BACKGROUND: The yeast Candida is one of the most frequent pathogens isolated from bloodstream infections and is associated with significant morbidity and mortality. Problems with clinical and microbiological diagnosis of invasive candidiasis (IC) have prompted the development of non-culture-based laboratory methods. Previous reports suggest that serological detection of antibodies might be useful for diagnosing systemic candidiasis. METHODS: Diagnosis of IC using antibodies against recombinant Candida albicans enolase (Eno) and fructose-bisphosphate aldolase (Fba1) was evaluated. Using recombinant Eno and Fba1 as coating antigens, enzyme-linked immunosorbent assays (ELISAs) were used to analyze sera from patients with candidemia (n = 101), Candida colonization (n = 50), bacteremia (n = 84), invasive aspergillosis (n = 40); and from healthy controls (n = 200). RESULTS: The results demonstrated that ELISA detection of anti-Eno and anti-Fba1 IgG distinguished IC from other pathogenic infections in patients and healthy individuals. The sensitivity, specificity, and positive and negative predictive values were 72.3%, 94.7%, 78.5% and 93% for anti-Eno, and 87.1%, 92.8%, 76.5% and 96.4% for anti-Fba1 antibodies, respectively. Combining these two tests improved sensitivity up to 90.1% and negative predictive value up to 97.1%, with specificity and positive predictive values of 90.6% and 72.2%. The tests were specific to the Candida genus and antibody titers were higher for candidemia patients than for controls. Positive antibody tests were obtained before blood culture results for 42.2% of patients for anti-Eno and 51.1% for anti-Fba1. CONCLUSION: These data suggest that tests that detect IgG antibodies against Candida enolase and fructose-bisphosphate aldolase, especially when used in combination, could be a powerful tool for diagnosing IC.

Surveillance study of species distribution, antifungal susceptibility and mortality of nosocomial candidemia in a tertiary care hospital in China.

BACKGROUND: Bloodstream infections due to Candida species cause significant morbidity and mortality, and the epidemiology of Candida infection is changing. Surveillance for candidemia is necessary to detect trends in species distribution and antifungal resistance. METHODS: The medical and electronic records of all patients who had candidemia at the authors' hospital from 2009 to 2011 were reviewed for demographic data and clinical information, including the infecting Candida species, resistance to antifungals and survival, and the presence of risk factors associated with candidemia. RESULTS: A total of 133 distinct episodes of candidemia were identified over the study period. The annual incidence of candidemia ranged between 0.71 and 0.85 cases/1000 hospital discharges. The most frequent Candida species were C. tropicalis (28.6%), followed by C. albicans (23.3%) and C. parapsilosis (19.5%). The rates of susceptibility to antifungal agents were as followed: voriconazole (97.8%), itraconazole (69.5%), fluconazole (46.1%), ketoconazole (38.9%). Out of 131 evaluable patients, 34 (26.0%) died within 30 days from the onset of candidemia. C. tropicalis candidemia was associated with the highest mortality rate (44.7%). Regarding the crude mortality in the different units, patients in Hemato-Oncology ward had the highest mortality rate (66.7%), followed by patients in cardiovascular wards and ICU (57.1% and 25.6%, respectively). Predictors of 30-day mortality were identified by uni- and multivariate analyses. Complicated abdominal surgery, presence of central venous catheter (CVC), neutropenia, candidemia due to C. tropicalis and poor treatment with fluconazole were significantly associated with the 30-day mortality. Presence of CVC (odds ratio[OR] = 4.177; 95% confidence interval [CI] = 1.698 to 10.278; P = 0.002) was the only independent predictor for mortality in the multivariate analysis. CONCLUSION: This report provides baseline data for future epidemiological and susceptibility studies and for the mortality rates associated with candidemia in our hospital. The knowledge of the local epidemiological trends in Candida species isolated in blood cultures is important to guide therapeutic choices.

Immunoproteomics based identification of thioredoxin reductase GliT and novel Aspergillus fumigatus antigens for serologic diagnosis of invasive aspergillosis.

BACKGROUND: There has been a rising incidence of invasive aspergillosis (IA) in critically ill patients, even in the absence of an apparent predisposing immunodeficiency. The diagnosis of IA is difficult because clinical signs are not sensitive and specific, and serum galactomannan has relatively low sensitivity in this group of patients. Therefore, more prompt and accurate disease markers for early diagnosis are needed. To establish disease markers demands a thorough knowledge of fungal antigens which may be detected in the serum or other body fluids of patients. Herein we report novel immunodominant antigens identified from extracellular proteins of Aspergillus fumigatus. RESULTS: Extracellular proteins of A. fumigatus were separated by two-dimensional electrophoresis (2-DE) and probed with the sera from critically ill patients with proven IA. The immunoreactive protein spots were identified by MALDI-TOF mass spectrometry (MALDI-TOF -MS). Forty spots from 2DE gels were detected and 17 different proteins were identified as immunogenic in humans. Function annotation revealed that most of these proteins were metabolic enzymes involved in carbohydrate, fatty acid, amino acid, and energy metabolism. One of the proteins, thioredoxin reductase GliT (TR), which showed the best immunoactivity, was analyzed further for secretory signals, protein localization, and homology. The results indicated that TR is a secretory protein with a signal sequence exhibiting a high probability for secretion. Furthermore, TR did not match any human proteins, and had low homology with most other fungi. The recombinant TR was recognized by the sera of all proven IA patients with different underlying diseases in this study. CONCLUSIONS: The immunoreactive proteins identified in this study may be helpful for the diagnosis of IA in critically ill patients. Our results indicate that TR and other immunodominant antigens have potential as biomarkers for the serologic diagnosis of invasive aspergillosis.

Corrigendum: Exercise and/or Cold Exposure Alters the Gene Expression Profile in the Fat Body and Changes the Heart Function in Drosophila.

[This corrects the article DOI: 10.3389/fendo.2022.790414.].

Exercise and/or Cold Exposure Alters the Gene Expression Profile in the Fat Body and Changes the Heart Function in Drosophila.

The major reason of human morbidity and mortality is obesity and related diseases. Brown adipose tissue (BAT) is associated with low total adipose tissue content and a lower risk of type 2 diabetes mellitus. Studies have shown that exercise and cold expose may induce browning. In this study, we verified (1) whether exercise and/or cold exposure can improve the expression level of ucp4c, serca, ampkα, camkII, sirt1, octß3r, and hamlet; (2) if these interventions can save cardiac dysfunction induced by a high-fat diet (HFD) in Drosophila. w1118 (wild-type) virgin female flies collected within 8 h after eclosion were divided into eight groups: the normal feed control group (NFD-C), the normal feed exercise group (NFD-E), the normal feed cold exposure group (NFD-CA), the normal feed exercise/cold exposure group (NFD-EC), the HFD control group (HFD-C), the HFD exercise group (HFD-E), the HFD cold exposure group (HFD-CA), and the HFD exercise/cold exposure group (HFD-EC). After exercise and/or cold exposure for 7 days, the mRNA expression levels of ucp4c, serca, ampkα, camk II, sirt1, octß3r, and hamlet were tested by qRT-PCR, and m-mode was used to assess cardiac function. In addition, we assessed the triacylglycerol (TAG) levels, motor ability, fat mass (by Oil Red O [ORO] staining), and morphological features. The results of TAG, ORO staining, and morphological features all indicate that after interventions, body size of Drosophila was smaller compared with the control group, irrespective of the feeding patterns. The mRNA expression levels of ucp4c, serca, octß3r, hamlet, ampkα, camkII, and sirt1 were changed to varying degrees under different intervention states (exercise and/or cold exposure). Cold exposure and exercise/cold exposure partly improved cardiac function and the normal fruit flies' cardiac function and exercise ability. However, after exercise intervention, exercise ability and heart function were improved in both HFD and normal-fat diet (NFD) fruit flies. In conclusion, different intervention states (exercise and/or cold exposure) can change the mRNA expression levels of ucp4c, serca, octß3r, hamlet, ampkα, camkII, and sirt1. Exercise is the most effective way to restore HFD-induced cardiac dysfunction.

Sperm protein 17 is highly expressed in endometrial and cervical cancers.

BACKGROUND: Sperm protein 17 (Sp17) is a highly conserved mammalian protein in the testis and spermatozoa and has been characterized as a tumor-associated antigen in a variety of human malignancies. Many studies have examined the role of Sp17 in tumorigenesis and the migration of malignant cells. It has been proposed as a useful target for tumor-vaccine strategies and a novel marker to define tumor subsets and predict drug response. This study aimed to investigate the expression of Sp17 in endometrial and cervical cancer specimens, its possible correlation with the pathological characteristics, and its value in the diagnosis and immunotherapy of the related cancers. METHODS: The monoclonal antibodies against human Sp17 were produced as reagents for the analysis and immunohistochemistry was used to study two major kinds of paraffin-embedded gynecological cancer specimens, including 50 cases of endometrial cancer (44 adenous and 6 adenosquamous) and 31 cases of cervical cancer (15 adenous and 16 squamous). Normal peripheral endometrial and cervical tissues were used as controls. RESULTS: Sp17 was found in 66% (33/50) of the patients with endometrial cancer and 61% (19/31) of those with cervical cancer. Its expression was found in a heterogeneous pattern in the cancer tissues. The expression was not correlated with the histological subtype and grade of malignancy, but the staining patterns were different in endometrial and cervical cancers. The hyperplastic glands were positive for Sp17 in the normal peripheral endometrial and cervical tissues in 10% (8/81) of the patients. CONCLUSIONS: Sp17 is highly expressed in human endometrial and cervical cancers in a heterogeneous pattern. Although the expression frequency of Sp17 is not correlated with the histological subtype, the staining pattern may help to define endometrial and cervical cancers. Sp17 targeted immunotherapy of tumors needs more accurate validation.

Overexpression of human sperm protein 17 increases migration and decreases the chemosensitivity of human epithelial ovarian cancer cells.

BACKGROUND: Most deaths from ovarian cancer are due to metastases that are resistant to conventional therapies. But the factors that regulate the metastatic process and chemoresistance of ovarian cancer are poorly understood. In the current study, we investigated the aberrant expression of human sperm protein 17 (HSp17) in human epithelial ovarian cancer cells and tried to analyze its influences on the cell behaviors like migration and chemoresistance. METHODS: Immunohistochemistry and immunocytochemistry were used to identify HSp17 in paraffin embedded ovarian malignant tumor specimens and peritoneal metastatic malignant cells. Then we examined the effect of HSp17 overexpression on the proliferation, migration, and chemoresistance of ovarian cancer cells to carboplatin and cisplatin in a human ovarian carcinoma cell line, HO8910. RESULTS: We found that HSp17 was aberrantly expressed in 43% (30/70) of the patients with primary epithelial ovarian carcinomas, and in all of the metastatic cancer cells of ascites from 8 patients. The Sp17 expression was also detected in the metastatic lesions the same as in ovarian lesions. None of the 7 non-epithelial tumors primarily developed in the ovaries was immunopositive for HSp17. Overexpression of HSp17 increased the migration but decreased the chemosensitivity of ovarian carcinoma cells to carboplatin and cisplatin. CONCLUSION: HSp17 is aberrantly expressed in a significant proportion of epithelial ovarian carcinomas. Our results strongly suggest that HSp17 plays a role in metastatic disease and resistance of epithelial ovarian carcinoma to chemotherapy.

[Aberrant expression of sperm protein 17 enhances migration of ovarian cancer cell line HO-8910].

OBJECTIVE: To explore the effects of aberrant expression of sperm protein 17 (Sp17) on the migration of the ovarian cancer cell line HO-8910. METHODS: The recombinant plasmid pEGFP-Sp17 containing Sp17 and enhanced green fluorescent protein gene was transfected into the human ovarian cancer cell line HO-8910 with Lipofectamine 2000. The expression of Sp17 was examined by RT-PCR and Western blot, and the cell migratory capability detected by Transwell chamber assays. RESULTS: Sp17 was expressed as a fusion protein with EGFP after transfected. There was a significant difference in the migratory cell number of the transfected and the control cells (156.6 +/- 14.9/HP vs 39.3 +/- 8.53/HP, P < 0.05). CONCLUSION: The aberrant expression of Sp17 greatly enhances the migration of ovarian cancer cells.

[Detection of anti-Sp17 antibodies in infertile patients' serum and its clinical significance].

OBJECTIVE: To test anti-Sp17 antibodies in the serum of AsAb positive infertile patients, to investigate the proportion of anti-Spl7 antibodies in AsAb and their potential application to the serologic diagnosis of immune infertility and immunocontraception. METHODS: With human recombinant Sp17 as the antigen, the ELISA method was used to detect the positive rate, antibody titre and content of anti-Sp17 antibodies in the AsAb positive serum. RESULTS: The positive rate of anti-Sp17 antibodies in the AsAb positive serum was 56.5%, with no significant difference in the gender aspect. The percentage of anti-Sp17 antibodies in AsAb was (10.09 +/-7.45) %, with statistical significance (P <0.05). CONCLUSION: Sp17 is an important sperm antigen. Anti-Sp17 antibodies in the serum can be taken as auxiliary diagnostic index of infertility, and Sp17 is shown to be a potential candidate immunocontraception vaccine.

[Development of radioimmunoassay for human sperm protein 17].

OBJECTIVE: To develop a method of radioimmunoassay for human sperm protein 17(Sp17) determination. METHODS: Anti-recombinant human Sp17 antibody was prepared, the labeling of 125I-rhSp17 performed by chloramine T method, and radioimmunoassay of Sp17 developed. RESULTS: The assay range was 3.3 to approximately 800 microg/L, the sensitivity was 2.0 microg/L, and the intra- and inter-assay coefficients of variation (CV) were 7.5% to approximately 9.8% and 8.2% to approximately 13.2%, respectively. The serum Sp17 level in normal subjects was (15.60 +/- 7.66) microg/L (n = 59). CONCLUSION: This radioimmunoassay of Sp17 fulfills the reasonable requirements of clinical routine and scientific studies in terms of specificity, sensitivity and practicability. Measurement of Sp17 concentration is useful for assessing its native distribution and aberrant expression.

Anti-Sp17 monoclonal antibody-doxorubicin conjugates as molecularly targeted chemotherapy for ovarian carcinoma.

Sperm protein 17 (Sp17) is selectively overexpressed in several human malignancies including ovarian carcinoma, but is absent or expressed at low levels in most normal tissues. Previous work from our group characterized an anti-Sp17 monoclonal antibody (clone 3C12) and showed that it specifically targeted tumor cells. In this report, we investigated whether a novel immunoconjugate containing 3C12 linked to the chemotherapeutic agent doxorubicin [(DOX) Adriamycin] had antitumor activity against ovarian cancer cell lines and tumor models. DOX was conjugated to 3C12 using a linker, and the specificity of 3C12-DOX was examined in Sp17-positive SKOV3 and Sp17-negative COC2 ovarian cancer cells using cell-based ELISA and internalization assays. The cytotoxicity of 3C12-DOX was assessed with the MTT assay, and its therapeutic effectiveness was evaluated in immunodeficient mice bearing SKOV3 cells. In vitro, the 3C12-DOX immunoconjugate specifically bound to and was internalized by Sp17-positive SKOV3 cells but did not bind to Sp17-negative cells. Treatment with 3C12-DOX (0.001 to 10 µg/mL) decreased the viability of SKOV3 cells in a Sp17-specific manner. In vivo, 3C12-DOX (3 mg/kg) induced the regression of established SKOV3 xenograft tumors in BALB/c mice compared with control treatment. The antitumor effects of 3C12-DOX were significantly associated with the induction of apoptosis in tumor cells. In addition, 3C12-DOX showed no observable adverse effects or toxicity when compared with DOX alone in mice bearing ovarian tumor xenografts. Our findings suggest that 3C12-DOX may be a potential antibody-drug conjugate for clinical development.

Sperm protein 17, MAGE-C1 and NY-ESO-1 in hepatocellular carcinoma: expression frequency and their correlation with clinical parameters.

UNLABELLED: This study is dedicated to investigate the expression patterns of sperm protein 17 (Sp17), melanoma-specific antigen (MAGE)-C1 and New York esophageal squamous cell carcinoma-1 (NY-ESO-1), to explore the correlation between these cancer-testis antigens and clinical parameters, and to evaluate their values in diagnosis and differentiation of hepatocellular carcinoma. METHODS: Immunohistochemical staining was performed in 45 paraffin-embedded hepatocellular carcinoma specimens. 45 normal peripheral hepatic tissues collected from adjacent non-cancerous areas were used as controls. RESULTS: Positive results of immunohistostaining were obtained in 16 (35.6%), 7 (15.6%) and 36 (80.0%) samples using MAGE-C1, NY-ESO-1 and Sp17 antibodies, respectively. The immunoreactivity of Sp17 was also found in 7 (14.0%) control samples. A statistical correlation between the frequency of Sp17 expression and tumor differentiation grade in hepatocellular carcinoma was confirmed. CONCLUSIONS: Sp17 is highly expressed in hepatocellular carcinoma cells. The frequency of Sp17 expression is closely related to the pathologic differentiation in hepatocellular carcinoma.

Anti-Sp17 monoclonal antibody with antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity activities against human ovarian cancer cells.

Sperm protein 17 (Sp17) is a cancer testis antigen that has been shown to be overexpressed in a variety of gynecologic malignancies, in particular ovarian cancer. Emerging evidences indicate that Sp17 is involved in tumorigenesis and in the migration of malignant cells. It has been proposed as a useful target for tumor-vaccine strategies and a novel marker to define tumor subsets and predict drug response. However, the antitumor activity of anti-Sp17 monoclonal antibody (anti-Sp17 mAb) has not been investigated. In this study, the in vitro cytotoxicity, antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) activities of anti-Sp17 mAb were evaluated using Sp17-positive ovarian cancer cells as targets, Sp17-negative ovarian cancer cells as the control, and healthy human peripheral blood monocytes and healthy human serum as effectors. Our preliminary results indicate that the direct cytotoxicity of anti-Sp17 mAb against the investigated ovarian cancer cells was very weak. However, the cytotoxicity of anti-Sp17 mAb, mediated by peripheral blood mononuclear cells (PBMCs), as ADCC, or by human serum, as CDC, was relatively strong in the Sp17-positive ovarian cancer cells. This finding suggested that anti-Sp17 mAb could be a useful tool against ovarian cancer and may provide insight into the development of low side-effect targeting therapy for this malignant disease.

Antibody specific to thioredoxin reductase as a new biomarker for serodiagnosis of invasive aspergillosis in non-neutropenic patients.

BACKGROUND: Invasive aspergillosis (IA) is an important cause of mortality in critically ill patients, but the diagnosis is difficult as clinical and radiological signs are neither sensitive nor specific. Serum galactomannan (GM) is a useful marker for IA, but exhibits low sensitivity in non-neutropenic patients. In our previous work, strong antibody reactivity to thioredoxin reductase of Aspergillus fumigatus was found in non-neutropenic IA patients. METHODS AND RESULTS: Using recombinant thioredoxin reductase GliT (TR), an antigenic protein secreted by A. fumigatus, as the coating antigen, an enzyme-linked immunosorbent assay (ELISA) for detecting anti-TR antibodies was developed. The antibody response to TR in IA animal models and 42 non-neutropenic patients with culture- and/or histology-documented IA was investigated. The results showed that anti-TR antibody was detectable in rabbit serum 7-9 days after exposure to the fungus. The sensitivity and specificity of the anti-TR antibody assay in patients were 80.9% and 96%, respectively, while the sensitivity of GM in this group of patients was only 52.3%. The specificity of the assay was confirmed by testing the sera from patients infected with other pathogenic fungal species and bacteria.

In vivo molecular targeting effects of anti-Sp17- ICG-Der-02 on hepatocellular carcinoma evaluated by an optical imaging system.

BACKGROUND: As the expression of human sperm protein 17 (Sp17) in normal tissue is limited and the function is obscure, its aberrant expression in malignant tumors makes it to be a candidated molecular marker for tumor imaging diagnosis and targeting therapy of the diseases.The aim of this research is to evaluate the targeting effects of anti-sperm protein 17 monoclonal antibody (anti-Sp17) on cancer in vivo and investigate its usefulness as a reagent for molecular imaging diagnosis. METHODS: Immunohistochemistry was used to identify the expression of Sp17 in a hepatocellular carcinoma cell line and tumor xenograft specimens. A near infrared fluorescence dye, ICG-Der-02, was covalently linked to anti-Sp17 for in vivo imaging. The immuno-activity of the anti-Sp17-ICG-Der-02 complex was tested in vitro by ELISA; it was then injected into tumor-bearing nude mice through the caudal vein to evaluate its tumor targeting effect by near infrared imaging system. RESULTS: Overexpression of Sp17 on the surface of the hepatocellular carcinoma cell line SMMC-7721 was demonstrated. Anti-Sp17-ICG-Der-02 with immuno-activity was successfully synthesized. The immuno-activity and photo stability of anti-Sp17- ICG-Der-02 showed good targeting capability for Sp17 expressing tumor models (SMMC-7721) in vivo, and its accumulation in the tumor lasted for at least 7 days. CONCLUSIONS: Anti-Sp17 antibody targeted and accumulated in Sp17 positive tumors in vivo, which demonstrated its capability of serving as a diagnostic reagent.

[Biological activity assays and cellular imaging of anti-human sperm protein 17 immunomagnetic nanoparticles].

AIM: To prepare anti-Sperm protein 17 (Sp17) immunomagnetic nanoparticles (IMNPs), and make foundation for target diagnosis of ovarian cancer by magnetic resonance imaging. METHODS: The anti-human Sp17 IMNPs were prepared by grafting anti-Sp17 antibodies on the surface of chitosan-coated magnetic nanoparticles (MNPs) using the linker of EDC/NHS (1-ethyl-3-[3-dimethylaminopropyl]carbodiimide/N-hydroxysuccinimide). The morphology and properties of the nanoparticles were characterized by transmission electronic microscopy (TEM), the conjugation of the antibodies was evaluated by native-polyacrylamide gel electrophoresis, the immunologic activity of IMNPs was evaluated by enzyme linked immunosorbent assay (ELISA). A set of in vitro magnetic resonance imaging (MRI) experiments were performed after incubated the IMNPs with human Sp17 gene transfected ovarian cancer HO-8910 cells. RESULTS: We had successfully grafted the MNPs with anti-Sp17 antibody and the IMNPs kept good bioactivity. The MRI showed that the IMNPs were targeted successfully to the positive cells, and no obviously non-specific adsorption was observed. CONCLUSION: The anti-Sp17 IMNPs with good specificity can used for further study of ovarian cancer target therapy.

[Preparation and characterization of the monoclonal antibody against human sperm protein 17].

AIM: To prepare and characterize the monoclonal antibody (mAb) against sperm protein 17 (Sp17). METHODS: Human Sp17 cDNA was cloned and recombinant Sp17 protein was expressed as a fusion protein with an N-terminal 6-His tag. The purified recombinant Sp17 protein was used to immunize BALB/c mice for preparing mAb. mAb-produced hybridoma cells were screened by ELISA and the specificity of mAb was analyzed by immunohistochemical staining and blocking test. RESULTS: Two hybridoma cells (3C12 and 3D6) secreting mAb against Sp17 were developed. The isotypes of these two mAbs, 3C12 and 3D6, were IgG(1) and IgM, respectively. ELISA detection showed that titers of mAbs, 3C12 and 3D6, were 1:64, 1:32 in cultured supernatant and 1:1 x 10(5), 1:5 x 10(4) in ascites, respectively. The results of immunohistochemical staining and blocking test indicated that 2 mAb specifically bound to Sp17 in human and rat testis and human ejaculated spermatozoa. Anti-Sp17 mAb also detected Sp17 in ovarian cancer. CONCLUSION: The successful preparation of anti-Sp17 mAb will be useful for assessing the native distribution and aberrant expression of Sp17 protein.

[Isolation of differentially expressed cDNA sequences in human gastric carcinoma by cDNA microarray].

BACKGROUND & OBJECTIVE: Some gastric carcinoma-related genes haven't been identified yet. The study was designed to isolate and identify differentially expressed cDNA sequences in gastric carcinoma, and further clone gastric carcinoma-related genes. METHODS: The differences of gene expression profile between 30 gastric carcinoma tissues and their adjacent normal tissues were analyzed by cDNA microarray which representing about 4 892 cDNA sequences, the differentially expressed cDNA sequences were analyzed by bioinformatics, and selected cDNA sequences were confirmed by reverse transcripase-polymerase chain reaction (RT-PCR). RESULTS: A total of 33 differentially expressed cDNA sequences were identified in gastric carcinoma, among which 18 up-regulated in gastric carcinoma,while 15 down-regulated. MDSCBC11 clone,represented a novel gene, located in chromosome band 1p35-36, and significantly down-regulated in 13 of 30 (43%) gastric carcinomas. CONCLUSIONS: Some gastric carcinoma-related cDNA sequences have been identified,they might be involved in pathogenesis of gastric carcinoma. This study provides a useful clue for further clone gastric carcinoma-related genes.