脂质运载蛋白型前列腺素D合酶在脑胶质瘤中的表达研究

目的　研究脂质运载蛋白型前列腺素D合酶 (L PGDS)mRNA及其蛋白在脑胶质瘤中的表达。方法　用实时荧光定量逆转录聚合酶链反应 (RT PCR)和免疫印记 (westernblot)检测L PGDSmRNA及其蛋白在脑胶质瘤组织和脑脊液中的表达水平。结果　实时荧光定量RT PCR显示 ,L PGDS的mRNA在 2 3例脑胶质瘤组织中的表达水平范围为 1 5× 10 3～ 7 0× 10 4 copy/μgRNA ,均值为 1 6×10 4 copy/μgRNA ,而在 5名正常组织中的表达水平均值为 8 1× 10 5copy/μgRNA。经统计学分析 ,两组间差异有显著性 (P <0 0 5 ) ,且表达水平与胶质瘤恶性程度相关。胶质瘤细胞株U 2 5 1的表达水平为4 7× 10 3copy/μgRNA。蛋白定量显示 ,脑胶质瘤患者脑脊液中L PGDS的吸光度比率均值为 0 47±0 12 ,而正常脑脊液中为 0 92± 0 2 6 ,两组间差异有非常显著性 (P <0 0 1)。结论　发现在脑胶质瘤中L PGDSmRNA及其蛋白表达水平下降 ,这可能对脑胶质瘤的诊断、治疗有潜在的应用价值。     

Expression of prostaglandin D synthase in ovarian cancer.

Lipocalin-type prostaglandin D synthase (L-PGDS) has recently been shown to be expressed in human brain tumors and breast tumors. However, L-PGDS expression has not been investigated in ovarian cancer. The objective of this study was to determine whether L-PGDS is expressed in human ovarian cancer. Lipocalin prostaglandin D synthase mRNA was cloned and sequenced by RT-PCR. Using in situ hybridization (ISH) technique, the expression of L-PGDS mRNA in 54 ovarian cancer was investigated. Expression of L-PGDS mRNA was found in tumor cells of all various types of ovarian cancers. Patterns of staining of tumor cells varied among different histological types of ovarian cancer. Significant discrepancy between the intensity of the staining and histological types of ovarian cancer could be established (p<0.01). It is reported for the first time that the expression of mRNA of L-PGDS exists in the ovarian cancer, and is related to the cancer type. This may have significance for the progress of ovarian cancer.     

Human glandular kallikrein in breast milk, amniotic fluid, and breast cyst fluid.

BACKGROUND: Human glandular kallikrein (hK2) belongs to the serine protease family of enzymes and has high sequence homology with prostate-specific antigen (PSA). The physiological role of hK2 has not as yet been determined, but there is evidence that it can regulate the proteolytic activity of PSA through processing and activating pro-PSA, an inactive precursor. Thus, it is conceivable that these two secreted proteins may coexist in biological fluids. Currently, hK2 is considered an androgen-regulated and prostate-specific protein. Recently, it has been demonstrated that hK2 is expressed in the breast cancer cell line T-47D after stimulation by steroid hormones, and we reported that hK2 can be detected in a subset of breast tumor extracts. These data suggest that hK2 may be expressed in tissues other than the prostate, such as those in which PSA has already been detected. Because hK2 is a secreted protein, it may be present in various biological fluids. METHODS: We analyzed milk samples from lactating women, amniotic fluid from pregnant women, and breast cyst fluid from patients with gross breast cystic disease, using a highly sensitive and specific immunoassay for hK2. RESULTS: hK2 was present in all three biological fluids. We suggest that the female breast may produce hK2 and provide evidence that hK2 may have value as an additional marker for the discrimination between type I and type II breast cysts. CONCLUSIONS: The female breast produces hK2 in addition to PSA. More studies are necessary to establish the role of this kallikrein in nondiseased breast, gross breast cystic disease, and breast cancer.     

Relationship between lipocalin-type prostaglandin D synthase and alpha-glucosidase in azoospermia seminal plasma

BACKGROUND: [corrected] To determine the correlation of lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase in semen. METHODS: We analyzed 68 seminal plasmas for lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase, L-PGDS was analyzed by ELISA. The semen donors were categorized in 3 groups: normal, obstructive and non-obstructive azoospermia. We then evaluated their correlation. RESULTS: The difference of L-PGDS concentration (P<0.001) and alpha-glucosidase activity (P<0.001) among the 3 clinical groups was statistically significant. Correlation between L-PGDS concentration and alpha-glucosidase was also statistically significant. L-PGDS concentration correlated positively with alpha-glucosidase activity (r=0.882). CONCLUSIONS: L-PGDS in seminal plasma, like alpha-glucosidase, suggests an obstruction of the seminal ducts and may be a potential marker that may aid in the differential diagnosis of obstructive and non-obstructive azoospermia.     

Development and evaluation of a practical ELISA for human urinary lipocalin-type prostaglandin D synthase

BACKGROUND: Urinary excretion of lipocalin-type prostaglandin D synthase (L-PGDS) is significantly increased in patients with chronic renal failure, but its diagnostic potential in less advanced stages of renal diseases remains to be elucidated. METHODS: Six mouse monoclonal antibodies (MAbs) were raised against recombinant human L-PGDS. We constructed a sandwich ELISA with two MAbs that recognized different epitopes with high affinities and assessed its assay performance and clinical utility with urine samples from healthy controls, diabetic patients, and patients with various renal diseases. RESULTS: Western blot analyses with NH(2)-terminus-truncated L-PGDS mapped the epitopes to Ala(23)-Val(28) (MAb-7F5 and -10A3), Ser(52)-Ala(73) (MAb-9A6), Tyr(107)-Val(120) (MAb-1B7 and -6F5), and Gly(140)-Pro(155) (MAb-6B9). A sandwich ELISA was constructed with MAb-1B7 and -7F5, the K(d) values of which were 3.6 and 3.9 nmol/L, respectively, for native L-PGDS. Recoveries were 91-111%, and intra- and interassay CVs were <6% and <9%, respectively. The ELISA showed parallelism of standard and urine samples and no significant interference by a variety of urinary constituents. Urinary L-PGDS excretion was significantly increased in patients with diabetic nephropathy, IgA nephropathy, and chronic glomerulonephritis even when serum creatinine was not increased. In patients with renal diseases, urinary L-PGDS was correlated with urinary albumin (r = 0.64; P <0.0001), N-acetyl-beta-D-glucosaminidase (r = 0.43; P <0.001), and serum creatinine (r = 0.66; P <0.0001). At a cutoff value of 284 mg/mol creatinine, the assay had sensitivities of 74% for diabetic nephropathy and 83% for chronic glomerulonephritis and a specificity of 93%. CONCLUSIONS: This ELISA system is suitable for measurement of urinary L-PGDS in a routine clinical assay and may be useful to detect less advanced stages of renal diseases.     

Relationships between androgen and estrogen sulfates in breast cyst fluid.

In 28 breast cyst fluids obtained from 20 patients (age 29-65 years) sodium, potassium and the sulfates (S) of estrone (E1), estradiol (E2), dehydroepiandrosterone (DHEA) and androsterone (A) were determined. The radioimmunoassays (RIA) used were validated for this particular biological fluid. According to electrolyte ratio (Na+/K+) the cyst fluids were subdivided into two groups: the first with low (less than 3) (n = 16) and the other with high (greater than 3) (n = 12) values. Markedly higher steroid sulfate levels were observed in the first group, the mean levels being: 147.7 nmol/l, 54.6 nmol/l, 108.1 mumol/l and 158.0 mumol/l for E1S, E2S, DHEAS and AS respectively. The mean levels in the second group were: 13.6 nmol/l, 6.7 nmol/l, 68.8 mumol/l and 33.6 mumol/l for E1S, E2S, DHEAS and AS, respectively. In the first group only E1S and E2S levels were significantly correlated (r = 0.51; P less than 0.05). Conversely, the steroid sulfate levels were significantly correlated with each other in the group with high electrolyte ratio. These data have confirmed preceding results and have clearly shown that breast cyst fluids with low electrolyte ratio contain more E2S than the other group. This finding might be correlated with the fact that patients with these breast cysts lined by with apocrine epithelium may be at a greater risk of breast cancer than those with the other type.     

Prostate specific antigen molecular forms in breast cyst fluid and serum of women with fibrocystic breast disease

We have analyzed matched serum and breast cyst fluid samples for total PSA from 148 patients with fibrocystic breast disease. We have also determined the molecular forms of PSA (free PSA and PSA bound to alpha1-antichymotrypsin) in 78 breast cyst fluid samples. We found that total PSA can be detected in all cyst fluids and in about 75% of female sera. The median total PSA concentration in breast cyst fluid (bcf) is about 30 times higher than the median in the corresponding sera. Breast cyst fluid and serum PSA are not correlated with each other. Total serum PSA is inversely associated with patient age but the inverse association between bcf PSA and age is weak. Lower total PSA in bcf was seen in women who breast feed, and higher bcf PSA is associated with multiple cysts. Type I cysts (with a high K+/ Na+ ratio) tend to have higher total PSA than Type II cysts. All but three of the fractionated cyst fluids (75/78; 96%) had free PSA as the predominant molecular form. The most consistent finding of our study was the positive association between the cyst fluid K+/Na+ ratio and the free to bound PSA ratio. This association was confirmed by Spearman correlation as well as by Wilcoxon and chi-square analysis. Secretory/apocrine cysts (Type I) tend to have more total PSA and proportionally more free PSA than transudative/flattened cysts (Type II).     

Lactate dehydrogenase, isoenzyme patterns and cation levels in human breast gross cyst fluid

In order to investigate the diagnostic and/or prognostic value of total activity and isoenzymatic patterns of lactate dehydrogenase, 24 human breast gross cystic fluids were studied. A comparison of total lactate dehydrogenase activity to the serum level revealed an increased activity in about 63% of the cases examined; moreover, a significant increase in the slow-moving lactate dehydrogenase 4 and 5 isoenzymes was observed in some cyst fluids. The levels of Na+ and K+ concentrations were also analyzed and two classes of cysts were identified: one presenting Na+ and K+ levels similar to those found in extracellular compartment; the other with high K+ and low Na+ levels, characteristic of an intracellular fluid. This latter pattern could indicate an active metabolism of the epithelial cells lining the cysts. This breast cyst fluid also showed increased levels of lactate dehydrogenase 4 and 5 isoenzymes. The correlation between an increased activity of lactate dehydrogenase 4 and 5 isoenzymes and high K+ and low Na+ levels could be the expression of a high biosynthetic activity and of an anaerobic metabolism in some cysts, suggesting the evolution of the breast gross cyst lesion to malignancy. The importance of these observations is discussed.     

Multi-task learning for segmentation and classification of tumors in 3D automated breast ultrasound images

Tumor classification and segmentation are two important tasks for computer-aided diagnosis (CAD) using 3D automated breast ultrasound (ABUS) images. However, they are challenging due to the significant shape variation of breast tumors and the fuzzy nature of ultrasound images (e.g., low contrast and signal to noise ratio). Considering the correlation between tumor classification and segmentation, we argue that learning these two tasks jointly is able to improve the outcomes of both tasks. In this paper, we propose a novel multi-task learning framework for joint segmentation and classification of tumors in ABUS images. The proposed framework consists of two sub-networks: an encoder-decoder network for segmentation and a light-weight multi-scale network for classification. To account for the fuzzy boundaries of tumors in ABUS images, our framework uses an iterative training strategy to refine feature maps with the help of probability maps obtained from previous iterations. Experimental results based on a clinical dataset of 170 3D ABUS volumes collected from 107 patients indicate that the proposed multi-task framework improves tumor segmentation and classification over the single-task learning counterparts.     

BI-RADS分级在乳腺单纯浸润性非特殊癌中的价值与应用

目的  分析BI-RADS标准化超声分级4C类病灶征象运用于乳腺浸润性癌非特殊型诊断时与病理征象的关联性及此类型癌的年龄分布,探讨BI-RADS分级标准应用于乳腺浸润性癌非特殊型诊断中的价值。方法  随机收集福建省立医院超声科于2020年5月~2023年5月所收治的88例超声BI-RADS 4C类的乳腺癌患者作为研究对象,针对患者年龄及病理检查结果进行分类,并对不同年龄段患不同类型乳腺疾病的患者的病灶超声特征与病理特征相关性进行讨论。结果  超声检查及病理追踪分析结果发现,在BI-RADS 4C类88例患者中,≥40岁的患者人数占90%, < 40岁的患者人数占10%;在乳腺浸润性非特殊癌中,≥40岁的患者人数占比95%, < 40岁的患者人数占比5%。单纯浸润性非特殊癌在病理检查结果中占总数的64.77%,其余非单纯浸润性非特殊癌占35.23%。结论  综合数据分析超声检查与BI-RADS分级结合在单纯浸润性非特殊癌中的检出率明显较高,有利于制定对应的治疗措施,更加突出BI-RADS分级在乳腺单纯浸润性非特殊癌中的检出率和运用。     

Concentrations of prostaglandin endoperoxide synthase and prostaglandin I2 synthase in the endothelium and smooth muscle of bovine aorta.

Platelets adhere to the subendothelial layer of newly deendothelialized arteries. Attachment can be reduced with exogenous prostacyclin (PGI2). Thus, the subendothelium may be unable to produce sufficient PGI2 to prevent platelet adherence and subsequent platelet-platelet interaction. Consistent with this explanation are data from an earlier report (1977. Moncada S., A. G. Herman, E. A. Higgs, and J. R. Vane. Thromb. Res. 11:323-344) indicating that the smooth muscle layer of aorta has only 10-15% of the capacity of endothelial cells to synthesize PGI2. We have measured the concentrations of PGI2 synthase and prostaglandin endoperoxide (PGH) synthase in bovine aorta and obtained results quite different from those described in this earlier report. Tandem immunoradiometric assays for PGI2 synthase and PGH synthase antigens were used to quantitate these proteins in detergent-solubilized homogenates of endothelial cells and smooth muscle tissue prepared from 10 different bovine aorta. The concentrations of PGI2 synthase in endothelial cells and smooth muscle were found to be the same. However, the concentration of PGH synthase in endothelial cells averaged greater than 20 times that of smooth muscle. Results similar to those determined by immunoradiometric assay were also obtained when PGH synthase and PGI2 synthase catalytic activities were measured in preparations of endothelial and smooth muscle cells. Furthermore, when bovine aorta and renal arteries were subjected to immunocytofluorescence staining using monoclonal antibodies to PGI2 synthase, fluorescence staining of equivalent intensity was detected in both the endothelial cells and the smooth muscle. Moreover, the intensity of fluorescence was similar throughout cross-sections of vascular smooth muscle, indicating that there is no gradient in PGI2 synthase concentrations between the endothelium and adventitia. Our results indicate that the propensity of platelets to adhere to the subendothelium of deendothelialized arteries and form aggregates cannot be attributed simply to an inability of the denuded vasculature to produce PGI2 from PGH2, but may be a consequence of the low PGH synthase activity of smooth muscle. Consistent with this concept are the results of Eldor et al. (1981. J. Clin. Invest. 67:735-741) who reported that increases in PGH synthase activity are associated with formation of a nonthrombogenic neointima.     

MR imaging: its current and potential utility in the diagnosis and management of breast cancer

The rapid evolution of the treatment of breast cancer has been paralleled by a similar rapid improvement in the imaging of breast cancer. High-resolution contrast-enhanced MR imaging of the breast has recently emerged as a sensitive instrument for the detection of breast cancer. The sensitivity of MR imaging makes it an excellent tool in specific clinical situations, such as the detection of local recurrence in patients who have received breast-conservation therapy. Furthermore, MR imaging of the breast has the potential to be a powerful aid in presurgical planning and to be a useful adjunct to mammography in selected patients. MR imaging, however, has a significant false-positive rate, is not readily available in all areas, and is more expensive than mammography and sonography. It also remains unclear if alterations of management plans based on MR imaging findings actually benefit affected patients. Therefore rigorous clinical trials are needed to define precisely the exact role that MR imaging should play in the diagnosis and management of breast cancer patients.     

The glomerular sieving of pepsinogen A and C in man

Pepsinogen A (PGA) and pepsinogen C (PGC) are negatively charged, low molecular weight (LMW) proteins with a striking difference in renal handling: PGA (molecular weight 43,500 daltons) shows a high fractional excretion while the fractional excretion of PGC (molecular weight 40,500 daltons) is low, presumably due to tubular reabsorption. As these data suggest a high glomerular sieving of pepsinogens, we assessed the glomerular sieving coefficient (GSC) of PGA, PGC and several other proteins from their renal extractions. For this purpose blood samples were obtained simultaneously from the aorta (A) and right renal vein (V) in nine patients undergoing an elective heart catheterization. After correction of A-V differences for diuresis with the A-V difference of transferrin, GSCs (+/- SEM) for PGA and PGC were 0.90 +/- 0.14 and 0.85 +/- 0.17, respectively, GSC of beta 2-microglobulin being 0.90 +/- 0.12. For albumin and IgG, known to have a low GSC, low values were found. It is concluded that the GSC of a LMW protein in man can be calculated from both its A-V difference over the kidney and the A-V difference of an inert marker with a GSC of 1, provided they are corrected by the A-V difference of an inert marker with a GSC of 0. Our results demonstrate that PGA and PGC are almost freely filtered through the glomerular basement membrane despite their size and negative net molecular charge.     

Technique for the primary culture of human breast cancer cells and measurement of their prostaglandin secretion.

1. A method is described for the primary culture of human breast tumour cells on feeder layers of the STO mouse embryo fibroblast cell line. 2. The secretion of the prostaglandins E2 and F2 alpha from the cells was measured and the results indicate that the secretion of both prostaglandins was dependent on oestrogen-receptor status, with cells from oestrogen-receptor-positive tumours secreting significantly more prostaglandin than cells from oestrogen-receptor-negative tumours. 3. Prostaglandin E2, but not prostaglandin F2 alpha, secretion was also significantly greater from cells of tumours from postmenopausal women than from cells of tumours from premenopausal women. Small (< 3 cm) tumours secreted significantly more prostaglandin than large (> 3 cm) tumours, and increased levels of prostaglandin were secreted with advancing clinical stage (T1-T4). 4. Additional evidence for increased prostaglandin metabolism in oestrogen-receptor-positive tumours compared with oestrogen-receptor-negative tumours was obtained from studies on the uptake of [14C]arachidonic acid from the cultures. Significantly more labelled arachidonic acid was incorporated into cells from oestrogen-receptor-positive tumours compared with oestrogen-receptor-negative tumours, with the subsequent release of more prostaglandin in response to various stimuli.     

Niemann-Pick disease type C: cataplexy and hypocretin in cerebrospinal fluid

Niemann-Pick disease type C (NPC) is an inherited lipid storage disorder, characterized by a defect in intracellular trafficking of exogenous cholesterol that leads to the lysosomal accumulation of unesterified cholesterol. We report a Japanese patient with NPC caused by a homozygous c.2974 G > T mutation of the NPC1 gene, which predicts a glycine (GGG) to tryptophan (TGG) change at codon 992 (designated as p.G992W). This is a well-known NPC1 gene mutation that causes a unique phenotype of NPC, which has been limited to a single Acadian ancestor in Nova Scotia, Canada. Our patient characteristically started presenting with cataplexy at the age of 9 years. Recent studies have shown reduced hypocretin-1 levels in the cerebrospinal fluid (CSF) of narcoleptic patients with cataplexy. In our patient, the level of hypocretin-1 was determined as moderately low, 174 pg/ml (normal, > 200 pg/ml). To date, CSF levels of hypocretin-1 have been determined by using an identical assay method in 7 cases of NPC, including our case. All of the NPC cases with cataplexy demonstrated low levels of CSF hypocretin-1, confirming the association of reduced CSF hypocretin-1 levels with cataplexy in NPC.     

CA-199在肝囊肿患者中的表达

目的:探讨血清及囊液中CA-199与肝囊肿的关系.方法:检测肝囊肿患者血清和囊液中CA-199浓度,对比经皮肝穿刺囊肿抽液并无水酒精注射前后血清中CA-199浓度变化.并分析其与囊腔直径、囊液量间的关系.结果:血清CA-199浓度升高的肝囊肿患者,经穿刺抽液并无水酒精注射后浓度明显下降[(51±3)U/mL vs (24±2)U/mL,P<0.05];血清CA-199浓度与肝囊肿囊腔直径及囊液量呈正相关.结论:血清CA-199有可能作为肝囊肿穿刺抽液并无水酒精硬化治疗后效果判断的指标之一.     

应用超声鉴别甲状腺囊肿囊液黏性及三通硬化治疗的优势

目的探讨超声判断甲状腺囊肿囊液黏稠度的效果及应用三通硬化治疗的优点。方法观察初步诊断为甲状腺囊肿的47例患者的超声影像特点,将其随机分为三通组（三通的接口连接静脉输液针和两个注射器针管）24例和传统组（只用注射器）23例进行囊肿穿刺,抽取囊液后注入1/3囊液量的无水乙醇（浓度〉95%）。比较超声影像和囊液的特点,并记录穿刺时间及不良反应。结果超声表现为囊液中有点状高回声伴声影且点状高回声不移动者均抽出稠厚液体,而囊液表现为液性暗区、无点状高回声或囊液有点状高回声向下坠落者抽出的均为稀薄液体。三通组的穿刺时间为（5.7±1.1）min,传统组为（7.2±1.3）min,两组比较差异有统计学意义（P〈0.05）。三通组无液体溢出或漏出,污染率0%;传统组污染率为36.8%,两组比较差异有统计学意义（P〈0.05）。结论通过超声检查可判断甲状腺囊肿囊液的黏稠度。应用三通硬化治疗操作简单、不良反应少。     

三阴性乳腺癌中间质-上皮细胞转化因子及脂肪酸合成酶的表达情况及对预后的影响

目的 观察间质-上皮细胞转化因子(c-Met)及脂肪酸合成酶(FASN)在三阴性乳腺癌患者中的表达情况及对预后的影响.方法 采用免疫组织化学染色法(SP)检测218例三阴性乳腺癌组织及癌旁组织中c-Met及FASN蛋白的表达水平,观察c-Met及FASN阳性表达情况,并探讨其与临床病理特征的关系及对预后的影响.结果 2...     

DPP-4 inhibitors and their potential role in the management of type 2 diabetes

The dipeptidyl peptidase 4 (DPP-4) inhibitors enhance the body's own ability to control blood glucose by increasing the active levels of incretin hormones in the body. Their mechanism of action is distinct from any existing class of oral glucose-lowering agents. They control elevated blood glucose by triggering pancreatic insulin secretion, suppressing pancreatic glucagon secretion, and signalling the liver to reduce glucose production. The leading DPP-4 inhibitors have shown clinically significant HbA1c reductions up to 1 year of treatment and offer many potential advantages over existing diabetes therapies including a low risk of hypoglycaemia, no effect on body weight, and the potential, based on animal and in vitro studies, for the regeneration and differentiation of pancreatic beta-cells. They are efficacious as monotherapy and also in combination with commonly prescribed antidiabetic agents and are suitable for once-daily oral dosing. Consequently, many DPP-4 inhibitors such as vildagliptin (Galvus; LAF-237), sitagliptin (Januvia; MK-0431), and saxagliptin (BMS-477118) have advanced into late-stage human clinical trials. Search strategy and selection criteria This review was built on a systematic MEDLINE search for publications on the subject with the key words: DPP-4 inhibitor; vildagliptin (LAF-237); sitagliptin (MK-0431); saxagliptin (BMS-477118); and type 2 diabetes; up to August 2006. Meeting abstracts were also searched, as much of the data currently only exists in abstract form. Take home message for clinician The DPP-4 inhibitors appear to have great potential for the treatment of type 2 diabetes, but time will tell if this will be realized. While they do not lower glucose to a greater extent than existing therapies, they offer many potential advantages, including the ability to achieve sustainable reductions in HbA1c with a well-tolerated agent that has a low risk of hypoglycaemia and no weight gain, and which can be administered as a once-daily oral dose.