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Immunostaining for detecting lymph node metastases in non-small cell lung cancer.

OBJECTIVE: To detect lymph node metastases by immunohistochemistry, where previously undetected by routine histopathology. DESIGN: Immunostaining was carried out for high- and low molecular weight cytokeratins, and Ber-EP4 in 19 consecutive lung cancer patients who had undergone systematic mediastinal lymph node dissection. RESULTS: Eleven (58%) epidermoid carcinomas, 6 (32%) adenocarcinomas, and 2 (10%) bronchiolo-alveolar carcinomas were detected. These included 4 (21%) stage IA carcinomas, 6 (32%) stage IB, 6 (32%) stage IIB, 1 (5%) stage IIIB and 2 (10%) stage IV. Immunostaining did not reveal any undetected metastases. Two patients (squamous cell carcinoma T1N0; adenocarcinoma T1N0) had metastases (skeletal; ipsilateral lung) at time of surgery, and one patient (squamous cell carcinoma T2N0) had a regional and systemic relapse 10 months later. Serial sectioning with immunostaining of the lymph nodes from these three patients was also negative. CONCLUSION: We conclude that, even with the use of immunostaining, negative lymph nodes will not assure a good prognosis, and different determinants probably exist for lymphatic and hematogenic metastases in non-small cell lung cancer.

Aged↗

Early detection of radiation-induced structural changes in rat trabecular bone.

OBJECTIVE: Early detection of bone changes following radiotherapy is very important. The aim of this study is to establish a radiographic image analysis method for early detection of radiation-induced trabecular bone changes. METHODS: Thirty-five Wistar rats were used for the experimental model of trabecular bone changes, which were induced by X-ray irradiation with 30 Gy to develop simulated osteomyelitis. Standardized 2.2 x direct magnification radiography was performed immediately prior to X-ray irradiation and once a week for 4 weeks following irradiation. The latent X-ray images were scanned using a computed radiography (CR) system. Mathematical morphological processing was then applied to the CR image data, which allowed the bone trabecular pattern features to be extracted as skeletal binary images. The sensitivity and specificity for detecting early trabecular bone changes in CR images and in skeletal binary images were evaluated. Quantitative analyses with quantum level value (QL value), skeletal pixel percentage (SKP) and star volume analysis (skeletal volume (Vsk), skeletal space volume (Vsp)) were performed. Histopathological examination was also conducted for confirmation of physical changes in the bone. RESULTS: Visual observation of the skeletal binary images provided a higher sensitivity than the CR images. In the quantitative analysis, SKP, Vsk and Vsp values provided higher sensitivity than QL values. CONCLUSION: The results of the present study suggest that a combined radiographic image analysis method using CR, mathematical morphological processing, SKP, Vsk and Vsp can be useful for the early detection of radiation-induced trabecular bone changes.

Analysis of Variance↗

Streaming detection for evaluation of indeterminate sonographic breast masses: a pilot study.

OBJECTIVE: Streaming detection is a novel sonography technique that uses ultrasonic energy to induce movement in cyst fluid that is detected on Doppler sonography. This pilot study evaluates the utility of streaming detection for differentiating cysts from solid masses in breast lesions that are indeterminate on sonography. SUBJECTS AND METHODS: Thirty-nine lesions-11 simple cysts and seven solid masses (control group) and 21 masses with indeterminate findings for the diagnosis of a cyst versus a solid lesion (study group)-in 34 patients were evaluated using streaming detection. All lesions underwent cyst aspiration or biopsy (n = 35) or were diagnosed simple cysts (n = 4) on sonography. Lesion size and depth were recorded. Streaming detection software was placed on conventional sonography units. Acoustic pulses were focused on the lesion, and if fluid movement was generated, it was seen on the spectral Doppler display as velocity away from the transducer. Lesions were then aspirated or underwent biopsy, and the viscosity of the aspirated fluid was recorded. The sensitivity and specificity of the technique and the effect of cyst size, cyst depth, and fluid viscosity in diagnosing fluid-filled cysts were assessed. RESULTS: Overall, 31 cysts and eight solid masses (seven benign, one carcinoma) were diagnosed in the study and control groups. Aspiration of indeterminate lesions resulted in 20 cysts and one solid mass. Lesions ranged in size from 4 to 47 mm and in depth from 4 to 29 mm. In the control group, streaming detection correctly showed nine of the 11 simple cysts (sensitivity, 82%; positive predictive value, 100%), and acoustic streaming was absent in all seven solid masses (specificity, 100%; negative predictive value, 78%). Of the indeterminate lesions, streaming detection allowed correct identification of 10 of 20 cysts (sensitivity, 50%; positive predictive value, 100%). Acoustic streaming was not detected in the one solid study group lesion. Neither cyst size or depth nor fluid viscosity had a significant effect on the ability to detect fluid. CONCLUSION: The streaming detection technique improved differentiation of cysts from solid masses in indeterminate lesions and has potential for reducing the number of recommended cyst aspirations for the diagnosis of indeterminate breast masses.

Adult↗

[Matched case-control study for detecting risk factors of breast cancer in women living in Chengdu].

OBJECTIVE: To detect the risk factors of breast cancer in women living in Chengdu in order to identify high risk population and to conduct proper interventions. METHODS: A matched case-control study was performed in 265 cases with breast cancer who lived in this area. RESULTS: In the univariate analysis, menarche to menopause period > or = 35 years, taking oral contraceptives, history of benign breast diseases, passive smoking, and syrup drinking are the statistically significant risk factors of breast cancer, whereas breast feeding and soybean food consumption are protective factors. Conditional logistic multivariate analysis revealed that history of benign breast diseases and passive smoking are risk factors while soybean food products and breast feeding are protective factors. CONCLUSION: The risk factors of breast cancer in women living in Chengdu are history of benign breast diseases and passive smoking.

Adolescent↗

Detection of genital colonization of group B streptococci during late pregnancy.

OBJECTIVE: To detect group B streptococcal carrier state of Saudi females during 3rd trimester of pregnancy and to assess type of specimens and the techniques used for the organism detection. METHODS: A total of 867 consecutive vaginal and rectal swabs were obtained from 217 pregnant women at > 28 weeks of gestation and their follow up testing from King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. Swab-specimens were cultured comparatively on Islam and Edwards blood agar plates, and into selective Lim broth. Enrichment Lim broth cultures (>12 hours) with and without positive modified coagglutination test were then subcultured on Islam and Edwards sheep blood agar plates. Presumptive colonies were then tested for group B streptococcus identity by convential biochemical reactions, serogrouping and serotyping. Collected neonatal swab-specimens (184) were also treated similarly. RESULTS: In comparison to Lim broth enrichment culture, the direct swab specimen culture on Edwards blood agar or Islam agar plates technique revealed 84% sensitivity and 100% specificity, whereas modified coagglutination test after selective Lim broth enrichment revealed 100% sensitivity and 96% specificity. Group B streptococcus was isolated in at least one of the specimens from the 217 patients in 66 cases. Of these 66 cases, group B streptococcus was isolated from both vaginal and rectal swabs in 33 (50%) cases and only from vaginal swabs in 22 (33%) and rectal swabs in 11 (17%) cases. Of the group B streptococcus positive cases, 10 (15%) cases had spontaneously lost their carriage, upon follow up testing, whereas out of the 151 negative cases, 4 (2.6%) cases became positive for group B streptococcus colonization upon follow up testing with an overall carriage rate of (60/217) 27.6%. Certain demographic factors were found to alter such rate of carriage. Additionally, 50% of group B streptococcal colonized mothers vertically transmitted the homologous serotypes of the organism to their newborns, but clinical infection was not recorded during the study period. CONCLUSION: Group B streptococci colonization rate among term Saudi pregnant women is relatively high (27.6%); and thereby constitutes a group of women whose infants are at great risk of early-onset invasive disease. The modified coagglutination test after growth amplification seems rapid and cost-effective to detect lightly or heavily group B streptococcal colonized women. Vaginal and rectal swab specimens at late pregnancy appeared necessary to accurately identify group B streptococcus maternal colonization.

Adolescent↗

[Detection of DNA damage of human sperm using single cell electrophoresis].

OBJECTIVES: To detect the sperm DNA damage and to evaluate its significance in male reproductive using single cell gel electrophoresis (SCGE). METHODS: Four hundred and eighteen sperm samples were analysed using the computer assisted analysis system and SCGE. The sperms samples were divided into five grades according to the extent of the sperm nuclear DNA damage. RESULTS: 1. When the sperm density is less than 20 x 10(6)/ml, the occurence of grade II and III are increased significantly; 2. In the unmotile grade d sperm the occurence of grade I comet amounts was 5.39%, the occurence of grade II and III was remarkably increased. There was a evidently variance between the grade d and grade a + b sperm. CONCLUSIONS: SCGE can be used to detect the sperm DNA breakage and to evaluate the sperm quality and damage.

Adolescent↗

[Detection of cholesterol ester transfer protein in semen of infertile patients].

OBJECTIVES: To detect the cholesterol ester transfer protein (CETP) levels in semen of infertile patients and evaluate the correlation between CETP in semial plasma and infertility. METHODS: One hundred and sixty-three infertile patients and fifteen fertile males were selected randomly. The routine examination of ejaculates was fulfilled by computer aided semen analysis (CASA). The CETP levels in all seminal plasma samples and fifty-five serum samples were detected by ELISA method. RESULTS: The CETP levels in infertile patients and fertile males were (2.21 +/- 1.23) microgram/L and (1.40 +/- 0.45) microgram/L, respectively. There were no significant differences between the two groups(P > 0.05). And there were no significant differences of CETP levels in seminal plasma among groups of azoospermia(n = 29), oligoasthenozoospermia (n = 58), oligospermia(n = 15), asthenozoospermia(n = 44) and normozoospermia(n = 17) in the infertile patients(P > 0.05). The CETP in seminal plasma and serum were detected in 55 infertile patients, and there was no correlation between CETP levels in seminal plasma and serum using Spearman analysis(r = 0.009, P > 0.05). The mean CETP level in seminal plasma was almost 1/1,000 of that in serum. CONCLUSIONS: The CETP level in seminal plasma is extremely low and has no relation with the changes of sperm density or motility. It may ensure the integrity of sperm membrane before the sperm enters into female genital tract.

Adult↗

Nested RT-PCR in detection of blood AFPmRNA in animal model of human hepatocellular carcinoma.

OBJECTIVES: To detect blood AFPmRNA in the nude mice bearing with human hepatocellular carcinoma (HCC) by using nested reverse transcriptase polymerase chain reaction (nested RT-PCR), and assess its significance in HCC distant metastasis. METHOD: We detected 20 blood samples from the nude mice bearing with human HCC by nested RT-PCR to find out AFPmRNA. RESULT: AFPmRNA was detected in 6 blood samples from the nude mice bearing with human HCC (30.0%), in which 4 mice developed distant metastasis. CONCLUSION: AFPmRNA may be used as an efficient and sensitive marker to detect blood spread of HCC cells. It can predict the occurrence of HCC distant metastasis.

Animals↗

[Study on the diagnostic significance of detecting the expression of AChR-gamma mRNA in rhabdomyosarcoma tissues].

OBJECTIVE: To detect over-expression of AChR-gamma mRNA in rhabdomyosarcoma tissues by duplex RT-PCR and discuss its potential in diagnosis of rhabdomyosarcoma. METHODS: Duplex RT-PCR was applied to the simultaneous detection of AChR-alpha and gamma subunit messenger RNA in 17 cases of rhabdomyosarcoma (9 ERMS, 6 ARMS, 2 PRMS). 20 cases of non-rhabdomyosarcomous small round cell tumors (6 poorly differentiated synovial sarcomas, 6 ES/PNET, 6 lymphomas, 2 neuroblastomas) and three normal muscle samples were also detected for AChR-alpha and gamma mRNA by the same method. RESULTS: AChR-alpha and AChR-gamma mRNA were expressed in all the cases of rhabdomyosarcoma. The rate of quantity in both transcripts was AChR-gamma/AChR-alpha >or= 1, but the rate for three normal muscle samples was < 1. Cases of non-rhabdomyosarcomous small round cell tumors were all negative for AChR-gamma. CONCLUSION: AChR-gamma mRNA expression detected by molecular genetic methods is useful in diagnosis and differential diagnosis of rhabdomyosarcoma.

Diagnosis, Differential↗

[Detection of the Salmonella invasion gene invA using molecular beacon probe].

OBJECTIVE: To detect Salmonella invasion gene invA. METHODS: The invA gene of Salmonella was amplified with PCR reaction system containing the molecular beacon probe labeled with 6-carboxy fluorescein (6-FAM) at its 5' end and with (4-dimethylaminophenylazo)benzoic acid (DABCYL) at its 3' end, and the fluorescence signal was read at the end of PCR. RESULTS: The fluorescence values of S.enteritidis, S. paratyphi A, S. paratyphi B, S.typhi antigen H, S.typhi, and enteroinvasive E.coli were 161.6, 104.5, 85.9, 83.1, 94.8 and 46.1 respectively. The results of PCR of Salmonella was consistent with that of agarose gel electrophoresis. CONCLUSION: PCR with fluorogenic molecular beacon probe is rapid, specific, sensitive, and convenient for detecting Salmonella with invA gene.

Bacterial Proteins↗

[Detection of porphyromonas gingivalis in buccal epithelial cells and subgingival plaque].

OBJECTIVE: To detect Porphyromonas gingivalis (P. gingivalis) in buccal epithelial cells and subgingival plaque from periodontally healthy subjects and patients with chronic periodontitis. METHODS: 40 subjects were included in the healthy group and 39 subjects were included in the diseased group in this study. Cells and subgingival plaque samples were collected. The extracted DNA was amplified with universal primers and P. gingivalis species-specific primer. RESULTS: P. gingivalis was detected in 37.5% of subgingival plaque samples and 32.5% of buccal mucosa samples in the healthy subjects, but 69.23% of subgingival plaque samples and 46.15% of buccal mucosa samples in the periodontitis group. Highly statistically significant differences were observed between healthy and periodontitis groups in the detections of P. gingivalis of subgingival plaque samples. CONCLUSION: P. gingivalis may be one of oral flora because it can be detected in the healthy population and not lead to destruction of supporting structures of the teeth.

Adult↗

[Detection of methylation of hMSH2 gene promoter region of esophageal cancer].

OBJECTIVE: To detect methylation in promoter region of hMSH2 gene in esophageal cancer. METHODS: Specimens of cancer and normal tissues freshly removed from 32 cases of esophageal cancer patients without previous radiotherapy, chemotherapy or other treatment were preserved at -80 degrees C within 30 min. Methylation specific PCR (MSP) was used to detect methylation of mismatch repair gene (MMR) hMSH2 in promoter region in esophageal cancer and normal esophageal tissues. RESULTS: The frequencies of methylation of hMSH2 gene in promoter region of cancer and normal esophageal tissues were 32.4% (11/32) and 0/30 (0%), respectively, and significant difference was found between the two groups (P < 0.01). The frequency of methylation in elder patients (> or = 70 years old) was significantly higher than that in younger patients (< 70 years old) (P < 0.05). Methylation was less frequently found in grade I-II (18.2%) than in grade III-IV (70.0%) (P < 0.05). CONCLUSION: Methylation of hMSH2 gene in promoter region is related to patients' age and histopathological grade of the esophageal cancer.

Aged↗

[Value of combinational detection by IgH and IgL primers in improving detection rate of lymphoma gene in paraffin-embedded tissue].

BACKGROUND & OBJECTIVE: Clonality detection through amplifying immunoglobulin heavy chain (IgH) by polymerase chain reaction (PCR) is a useful tool in diagnosis of lymphoma, but the false negative rate is high, especially in paraffin-embedded tissues. This study was to explore the value of tumor tissue microdissection and combinational detection of IgH and immunoglobulin light chain (Ig kappa or Ig lambda) in diagnosis of non-Hodgkin's lymphoma (NHL). METHODS: Two pairs of conventional primers for IgH and T-cell receptor gamma (TCRgamma), 2 novel designed pairs of primers for Ig kappa and Ig lambda were used to detect 58 paraffin-embedded blocks, which had been diagnosed by pathology and histochemistry. Of the 58 cases of lymph node tissues, 39 were B-cell lymphoma, 16 were T-cell lymphoma, and 3 were reactive proliferative lymph node tissue. Lymphoma cell lines DG75 and Jurkat were used as control. RESULTS: The positive rates of IgH primers (P1) and IgL primers (P kappa/P lambda) were 79.5% and 71.8% in the 39 cases of B-cell lymphoma (P>0.05), 6.3% and 12.5% in the 16 cases of T-cell lymphoma, respectively. The positive rate was greatly increased to 92.3% in combinationally detecting the primers for IgH and P kappa/P lambda. There was no positive detection among the reactive proliferative lymph node tissues. CONCLUSION: B-cell lymphoma detection rate can be significantly improved by the combination of IgH and IgL gene rearrangement primers, which provides efficient assistant method for the diagnosis and differential diagnosis of lymphoma.

DNA Primers↗

[Use of primary care services by adolescents and detection of health problems other than those for which they came].

OBJECTIVES: To detect in young people requesting Primary Care treatment the presence of hidden health problems common to this age-group and to evaluate the factors which may affect their being rarely mentioned. DESIGN: Descriptive study. PARTICIPANTS: Young people between 14 and 19 who attended two clinics at the Pola de Lena Health Centre from April to July 1994. The survey, using a personal interview, was aimed at the detection of acne, obesity, oral contraceptives, alcohol consumption and depressive disorders. MEASUREMENTS AND MAIN RESULTS: 138 young people (57 male and 81 female), with an average age of 16, requested care. 74.4% attended the consultation in company, although in 62.4% of cases the initiative was their own. The most common consultation was for acute infectious conditions (40.6%). 108 young people (78.3%) were interviewed, with 60 cases of acne, 25 of obesity and 28 possible depressive disorders being detected. We detected no cases of alcoholism or of uncontrolled oral contraception. For 64.3% of these problems, they had never consulted the doctor: they had consulted least for depressive disorders (3.6%). The commonest alleged reasons for not consulting were not being aware of the problem, indifference, ignorance of the condition's risks and consequences or of the existence of effective treatment. CONCLUSIONS: There are health problems of some importance about which young people requesting care do not consult. These problems should be actively detected and tackled, taking advantage of young people's attendance at the Health Centre for other reasons.

Acne Vulgaris↗

[Parental origin of extra chromosome 21 in Down syndrome detected by using short tandem repeat DNA polymorphisms after PCR amplification].

OBJECTIVE: To detect the parental origin of extra chromosome 21 in Down syndrome, using two (GT)n polymorphic markers-D21S215 and D21S120. METHODS: The alleles of D21S215 and D21S120 were amplified by polymerase chain reaction and identified with denaturing polyacrylamide gel electrophoresis followed with Ag-staining. The parental origin of extra chromosome 21 was determined by comparing genotypes of probands with their parents. RESULTS: The parental origin of extra chromosome 21 was determined in 17 Down syndrome out of 24, with 12 and 5 inherited from mother and father respectively. CONCLUSION: The parental origin of extra chromosome 21 in Down syndrome can be determined by analyzing the polymorphisms of D21S215 and D21S120. Etiological study of trisomy 21 should be focused on maternal meiosis.

Chromosomes, Human, Pair 21↗

[Detection of lead in environmental objects and biologic materials during monitoring].

The authors specified methods to detect lead in biologic materials. The quality control covered use of Russian and foreign standard samples, the results proved to agree. Lead was detected by AAS technique (direct and flow-type variants with preliminary concentration of lead) and ELRA method. The authors determined measurement limits sufficient for analysis of complex biologic materials. The measurement range for lead varied from 0.002 mg/kg (plants) to 3,000 mg/kg (soil); the range of serum lead levels was < 0.5-39 mg/dl.

Adult↗

Using statistical image models for objective evaluation of spot detection in two-dimensional gels.

Protein spot detection is central to the analysis of two-dimensional electrophoresis gel images. There are many commercially available packages, each implementing a protein spot detection algorithm. Despite this, there have been relatively few studies comparing the performance characteristics of the different packages. This is in part due to the fact that different packages employ different sets of user-adjustable parameters. It is also partly due to the fact that the images are complex. To carry out an evaluation, "ground truth" data specifying spot position, shape and intensities needs to be defined subjectively on selected test images. We address this problem by proposing a method of evaluation using synthetic images with unambiguous interpretation. The characteristics of the spots in the synthetic images are determined from statistical models of the shape, intensity, size, spread and location of real spot data. The distribution of parameters is described using a Gaussian mixture model obtained from training images. The synthetic images allow us to investigate the effects of individual image properties, such as signal-to-noise ratios and degree of spot overlap, by measuring quantifiable outcomes, e.g. accuracy of spot position, false positive and false negative detection. We illustrate the approach by carrying out quantitative evaluations of spot detection on a number of widely used analysis packages.

Algorithms↗