[Progress in the traceability of tumor marker detection].

Tumor markers (TM) detection is of great significance in tumor screening, monitoring and treatment intervention, which puts forward higher requirements for its detection quality. TM traceability is very important in the process of reagent production and clinical laboratory testing, which can help improving the reliability and comparability of TM testing. Based on the current principles and classification system of metrology traceability in the world, this paper reviews the quality requirements of reference materials and reference measurement methods related to protein and nucleic acid of TM, as well as the problems existing in the international convention reference measurement procedure and traceability system of TM, so as to provide a new idea for the quality assurance work of TM detection.

[The efficacy and safety of different antimicrobial regimens in carbapenem-resistant Klebsiella pneumoniae bloodstream infections].

Objective: To evaluate the efficacy and safety of different antimicrobial regimens in patients with bloodstream infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP). Methods: The clinical date of patients with CRKP bloodstream infections were retrospectively analyzed at the First Affiliated Hospital of Zhejiang University Medical College between January 2017 and January 2018. All subjects were separated into three groups based on antibiotics regimens over 72 hours, including meropenem 2.0 g every 8 hours, tigecycline 200 mg as initial dose and 100 mg every 12 hours, and polymyxin B 1.25 mg/kg every 12 hours as salvage treatment of tigecycline. Results: A total of 86 patients were finally recruited, including 14, 52 and 20 patients in groups of meropenem, tigecycline and polymyxin B salvage, respectively. All of the strains were resistant to meropenem and susceptible to tigecycline and polymyxin B initially, while 2 of them became resistant to tigecycline during treatment. The 28-day mortality was significantly higher in meropenem group (13/14) than that in tigecycline group and polymyxin B salvage group (61.5%, 32/52) and (12/20), respectively (P<0.01), while as no significant difference was seen in the last two groups (χ(2)=0.014, P>0.05). The incidences of hepatic impairment [3.8%(2/52) vs. 1/20] and renal dysfunction (0 vs. 1/20) between tigecycline group and polymyxin B salvage group were both comparable (P>0.05). Conclusion: The meropenem-based therapy is not recommended for CRKP-related bloodstream infections. Tigecycline-based therapy is still disappointing despite salvage use of polymyxin B after 72 hours. Hepatic and nephretic toxicities caused by additional polymyxin B are acceptable.

[Clinical characteristics of cardiomyopathies complicated with ventricular thrombosis].

Objective: To summarize the clinical characteristics of cardiomyopathy complicated with ventricular thrombosis. Methods: The clinical data of inpatients suffered from cardiomyopathy complicated with ventricular thrombosis in Fuwai Hospital between January 2015 and May 2019 were analyzed retrospectively. Results: A total of 125 cases were reviewed, and 24.8% were female. Dilated cardiomyopathy was the most common disease (62.4%), followed by arrhythmogenic right ventricular cardiomyopathy (ARVC) (13.6%) and hypertrophic cardiomyopathy (11.2%). There were 74.4% thrombosis in left ventricle, 12.8% in right ventricle and 12.8% in biventricle. The proportions of right ventricle thrombosis were higher in ARVC than in other cardiomyopathies (52.9% vs 6.5%, P<0.01). The majority suffered from cardiac function New York Heart Association (NYHA) Class Ⅲ (45.6%) and class Ⅳ (39.2%). The ratio of NYHA Class Ⅳ was higher in female patients than in male ones (25.8% vs 10.6%, P<0.05). In lab detection, positive results of D-Dimer and N terminal-pro B type natriuretic peptide (NT-proBNP) accounted for 72.8% and 97.6%, respectively. There were 2.5% patients died in the hospital or discharged because of the worsening of illness, the chances were higher in female than male patients (9.7% vs 0, P<0.01). Among these patients, one succumbed to massive ischemic stroke caused by ventricular thrombus detachment under standard anticoagulation therapy. Conclusions: Dilated cardiomyopathy is the most common cardiomyopathy complicated with ventricular thrombosis. The most common location of thrombosis is left ventricle. Right ventricle thrombosis is more common in ARVC. The majority suffer from moderate or severe cardiac dysfunction. Higer proportion of female patients suffer from anemia, severe condition and poor prognosis.

[The value of circulating tumor cells detected by chromosome centromere probe identification in diagnosis of non-small cell lung cancer].

Objective: This study explored the value of circulating tumor cells (CTC) detected by chromosome 8 centromere probe in diagnosis of non-small cell lung cancer (NSCLC) as well as correlation between CTC counts and clinical pathological characteristics. Methods: We collected 136 patients with newly diagnosed NSCLC (101 males and 35 females, age ranged from 34 to 79 years), 149 patients with non-malignant pulmonary diseases (103 males and 46 females, age ranged from 24 to 80 years) and 32 healthy volunteers (5 males and 27 females, age ranged from 24 to 42 years). Detection was performed using an epithelial cell adhesion molecule-independent strategy that combined immunocytochemistry staining (ICC) of CD(45) and fluorescence in situ hybridization detection (FISH) with chromosome 8 centromere probe (CEP8). CTC was defined as 4',6-diamidino-2-phenylindole (DAPI) positive, CD(45) negative and CEP8 more than 2 positive points. Quantitative data were reported as x±s and Mann Whitney test was used to compare them. Measurement data were analyzed as contingency tables and Pearson chi-squared test was used. P values of less than 0.05 were considered statistically significant. Results: CTCs were detected in 114 patients (83.8%) with NSCLC, 35 patients (23.5%) with non-malignant pulmonary diseases (P<0.000 1) and 5 volunteers (15.6%). CTC counts in NSCLC patients were (5.98±0.64) per 3.2 ml and (0.60±0.13) per 3.2 ml (P<0.000 1). A receiver operating characteristic curve (ROC) analysis showed similar capability of CTC count, with CEA, in discriminating NSCLC and non-malignant diseases with an area under ROC curve of 0.854 (95% confidence interval: 0.808-0.900, P<0.001). Cutoff of 2 circulating tumor cells per 3.2 ml peripheral blood gave the highest Youden Index of 0.614 in diagnosis of NSCLC with sensitivity of 72.1% (98/136) and specificity of 89.3% (133/149). When patients with CTC≥2/3.2 ml peripheral blood or serum CEA≥5 ng/ml were considered as having NSCLC, sensitivity and specificity of this combined test were 87.5% (119/136) and 85.9% (128/149), with a higher Youden Index of 0.734. No correlation was observed between positive rate of CTC (rate of patients with CTC≥2/3.2 ml peripheral blood) and age, gender, smoking status, pathologic types, and clinical stages. Conclusions: CTC counts detected by CD(45)-FISH is higher in NSCLC patients than in non-malignant disease patients. CTC≥2/3.2 ml peripheral blood is valuable in discriminating NSCLC from nonmalignant diseases, which can be more accurate when combined with CEA.

Submicrostructure and typing of female genital condylomata.

Thirty biopsies from female genital condylomata were examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to study structural characteristics and typing of condylomata. It was found that cytoplasmic clearing was marked in acuminate condylomata, diffuse interstitial and epithelial proliferation in nodular condylomata (flat condylomata), and invagination of the lesions into the interstitial tissue or glandular ducts in endophytic condylomata. In nodular condylomata, SEM also showed some structural features similar to those of intra-epithelial neoplasia. Microridges on the surface of squamous cells had villiform of granular changes. On the surface of a percentage of squamous or columnar cells, there were holes with a diameter of about 3 to 5 microns. A number of giant cells were seen among other cells. The cervical squamatization zone contained groups of special cells covered with dense microvilli. TEM of nodular condylomata revealed some pictures resembling active proliferation of tumor cells, such as enlarged or irregular nuclei (large N/C ratio), evaginated or invaginated nuclear membranes, condensed chromatin attached to the inner part of the nuclear membrane, transparent nucleoplasm, and frequent nucleosomes and karyokinesis. Virus particles with the morphological characteristics of HPV (naked hexagon-like particles with an average diameter of 45-50 nm) were seen in some nuclei with markedly condensed chromatin. It is suggested that HPV-induced genital condylomata, especially nodular one (flat condylomata), entail a potential progression to malignancy.

[Genetic mapping of the rice telomeric regions through PCR].

By using RAPD primer mediated asymmetric PCR (RM-PCR) method, a new type of molecular marker based on the telomeric repeats sequence was developed. PCR-based genetic mapping of telomeric repeat associated sequences (TASs) was conducted with a rice doubled haploid (DH) population derived from the inter-subspecific cross between indica variety (Zhaiyeqing8) and japonica variety (Jingxil7). Twenty-three loci were mapped onto the genetic map. Of these loci, 12 loci were mapped to the most distal position of eight chromosome arms and some of which may be located in subtelomeric region, five loci were mapped to the approximate positions of centromeric regions and six loci were mapped to other interstitial chromosomal regions.

The cycle of the seminiferous epithelium and stages in spermatogenesis in dd-mice.

For the basis to study the testicular functions using dd-mice, the cycle of the seminiferous epithelium and stages in spermatogenesis, classification of late spermatogenesis, and postnatal development of spermatogenesis were examined by light and electron microscopy in the testes from these animals at day of birth and 1, 2, 3, 4, 5, and 8 weeks of age. For light microscopy, paraffin sections of the testes were stained with periodic-acid and Schiff reagent (PAS) and hematoxylin. In spermiogenesis, 16 steps of spermatids were distinguished according to shapes of the nuclei and acrosomes. Step 15 and 16 spermatids, which have similar nuclei and acrosomes, were easily distinguishable by positions of nuclei and appearance of basophilic granules in the cytoplasm; steps 15 and 16 were thus subdivided into 3 and 2, respectively. The granules were accumulations of ribosome-like particles. Step 13 spermatids were present with step 1 spermatids, step 14 with steps 2 and 3, step 15 with steps 4-6, and step 16 with steps 7 and 8. Thus, the cycle of the seminiferous epithelium was divided into 12 stages (stages I-XII) in the order of steps 1 to 12. Type A spermatogonia were present in all stages, and intermediate and type B spermatogonia appeared from stages II and VI, respectively. In primary spermatocytes, the resting phase appeared from stage VII, leptotene from stage VIII, zygotene from stage X, pachytene from stage III, diplotene in stage XI, and diakinesis and secondary spermatocytes in stage XII. The seminiferous tubules were lined by Sertoli cells and contained gonocytes at day of birth, type A spermatogonia appeared at 1 week of age, spermatogenesis proceeded until pachytene spermatocytes at 2 weeks, until step 1 spermatids at 3 weeks, and all stages of the seminiferous epithelium were completed at 5 weeks. The findings indicate that combination of germ cells in each stage of the seminiferous epithelium in dd-mice does not correspond to generally accepted one, step 15 and 16 spermatids are accurately classified by referring the position of nuclei and appearance of granules in the cytoplasm, and spermatogenesis in immature mice proceeds faster than in adults.

Correlation of transformation from epithelial to mesenchymal-like morphology and endogenous bFGF levels in human nasopharyngeal carcinoma cells.

CG-1 human nasopharyngeal carcinoma cells in monolayer culture formed both cohesive, epithelial-like colonies and scattered, fibroblastic-like colonies in mixed proportions. In the presence of exogenously added bFGF (4 ng/ml), about 85% of the colonies formed were fibroblastic-like. CG-1 cells were capable of synthesizing and releasing bFGF, and, when compared by the immunological method, cells in fibroblastic-lke colonies were found to contain higher levels of endogenous bFGF than cells in the epithelial-like colonies. Furthermore, cells in the peripheral region of the epithelial-like colonies, which were fibroblastic-like in morphology, also appeared to contain higher levels of endogenous bFGF. In addition, in the presence of suramin, neutralizing antibody to bFGF, or neutralizing antibodies to bFGF and EGF, the number of cohesive colonies formed was greatly increased. Moreover, addition of the 2 M NaCl-eluted heparin-Sepharose fraction of the CG-1 cell-coditioned medium promoted the formation of dispersed colony in a dose-dependent manner. The results suggest that bFGF can regulate CG-1 cell phenotype in an autocrine manner.