[Analysis of problems and management of in vitro diagnostic reagent clinical trials in medical laboratory].

Clinical trials are an important method for evaluating the safety and efficacy of in vitro diagnostic reagents, and are a key basis for product registration review and approval. In order to strengthen the management of clinical trials of in vitro diagnostic reagents, the National Medical Products Administration and relevant departments have formulated a series of regulations at the regulatory level, and require applicants and clinical trial institutions to establish a quality management system for clinical trials of in vitro diagnostic reagents. Medical laboratory is the main department and implementer of in vitro diagnostic reagent clinical trials in medical institutions. In recent years, with the rapid development of the in vitro diagnostic industry, the clinical trial projects of in vitro diagnostic reagents conducted by medical laboratory have been increasing day by day. However, there are currently few discussions on the clinical trial of in vitro diagnostic reagents from the perspective of researchers. Therefore, this article summarizes the characteristics of clinical trials of in vitro diagnostic reagents, analyzes the problems and difficulties in conducting clinical trials of in vitro diagnostic reagents in current medical laboratories, and introduces the laboratory's experience in management; to provide reference for medical testing laboratories that have not yet conducted or have already conducted clinical trials of in vitro diagnostic reagents, in order to improve the quality and efficiency of clinical trials.

[Research progress of cell therapy in hereditary pulmonary alveolar proteinosis].

Hereditary pulmonary alveolar proteinosis (hPAP) is a rare interstitial lung disease caused by mutation in CSF2RA/CSF2RB, characterized by the deposition of pulmonary surfactant due to the alveolar macrophage dysfunction. The whole lung lavage can effectively alleviate the symptoms but is associated with potential complications. Cell therapy is a novel approach with advances that provide a new therapeutic strategy for the treatment of hPAP.

[Establishment and application of animal models of pulmonary alveolar proteinosis].

Pulmonary alveolar proteinosis (PAP) is a rare respiratory disease, but this disease has slow research progress. Animal model is an effective tool for basic research. Current PAP animal models are based on the main pathogenesis of granulocyte-macrophage colony stimulation factor (GM-CSF) signal disorder and environmental homeostasis imbalance in the alveoli. Application researches focus on the treatment strategies of PAP. The existing PAP animal models cannot fully reflect to the development of human PAP, which should be further developed and improved to provide the basis for clinical practice.

[Effect of serum potassium level on renal function progression in stage â ¢-â ¤ chronic kidney disease patients without dialysis].

Objective: To investigate the effect of serum potassium level and hyperkalemia on the renal function decline in chronic kidney disease (CKD) patients. Methods: The clinical data at baseline and follow-up in stage Ⅲ-Ⅴ CKD patients without dialysis who were followed up for more than one year in Tianjin First Central Hospital from May 2015 to June 2019 and Teikyo University School of Medicine from January 2008 to July 2013 were retrospectively collected. All patients were divided into stable group (337 cases), slow progression group (337 cases) and rapid progression group (338 cases) according to the tertile of estimated glomerular filtration rate (eGFR) slope (the annual average percentage of eGFR decline). Multivariate logistic regression analysis models were used to evaluate the correlations of baseline serum potassium or time-averaged serum potassium level with CKD rapid progression. Results: Three hundred and forty-three cases from Tianjin First Central Hospital and 669 cases from Teikyo University School of Medicine were included in the study, and 635 cases (62.7%) were male. The average age was (61±14) years old and the average eGFR decline slope was 4.0%/year. The levels of baseline serum potassium and time-averaged serum potassium of patients in the slow progression group [(4.47±0.52) and (4.51±0.43) mmo/L] and rapid progression group [(4.62±0.62) and (4.76±0.48) mmo/L] were higher than those in the stable group [(4.37±0.49) and (4.38±0.37) mmo/L] (both P<0.05). Meanwhile, 24.6% (83/338) of the patients in the rapid progression group had hyperkalemia at baseline (serum potassium ≥5.0 mmol/L) and 34.9% (118/338) of the patients had time-averaged serum potassium ≥5.0 mmol/L, which were higher than those in the stable group [10.7% (36/337) and 6.5% (22/337)] (both P<0.001). Multivariate logistic regression analysis showed that compared with the stable group, baseline serum potassium (OR=1.843, 95%CI: 1.051-3.234) and time-averaged serum potassium (OR=2.495, 95%CI: 1.040-5.987) were correlated with the rapid progression of CKD. Time-averaged serum potassium ≥5.0 mmol/L was the independent influencing factor for rapid progression of CKD. Conclusions: During the follow-up period, the average level of serum potassium in stage Ⅲ-Ⅴ CKD patients should be controlled under 5.0 mmol/L, which may reduce the risk of rapid decline of renal function.

Modeling of Li diffusion in nanocrystalline Li-Si anode material.

To quantify the Li diffusion behavior in nanocrystalline anode materials for lithium-ion batteries (LIBs), a hybrid model of the first principles calculation and diffusion kinetics was developed. The dependence of the Li diffusion on the electronic structure, solute concentration, grain size and temperature was described for the nanocrystalline Li-Si system. In contrast to conventional polycrystalline materials in which the activation barrier for Li diffusion decreases with the increase of concentration before amorphization, there exists a coordination effect of the solute concentration and grain size on the Li diffusion in nanocrystalline materials. A maximum diffusion coefficient can be obtained in the nanocrystalline Li-Si by a combination of the concentration and grain size, which is increased by two orders of magnitude from that in the coarse-grained counterpart. The present work advanced the understanding of the Li diffusion mechanisms during lithiation/delithiation of LIBs and may facilitate the development of nanocrystalline anode materials.

[The plasma level and gene expression in peripheral blood of interleukin-35 in patients with sarcoidosis and its clinical significance].

Objective: To investigate the expression of interleukin(IL)-35 protein and gene in peripheral blood of patients with sarcoidosis and its clinical significance. Methods: Peripheral blood samples from 98 patients with sarcoidosis and 98 healthy volunteers were collected at Peking Union Medical College Hospital between January 2016 and March 2017. The plasma levels of IL-35 were detected by enzyme-linked immunosorbent assays (ELISA), and the relationship between IL-35 and the clinical characteristics was analyzed. Real-time quantitative PCR was used to detect the expression levels of IL-35 subunit EBI3, p35 and T regulatory cell transcription factor Foxp3 mRNA in peripheral blood mononuclear cells, and their correlations were analyzed. Results: The plasma levels of IL-35 in patients with sarcoidosis (44±12) ng/L was significantly lower than that in the normal control group (55±12) ng/L (P<0.001). There was a positive correlation between the plasma levels of IL-35 and D(L)CO% predicted values (r=0.76, P<0.001), but it showed no significantly correlation with other clinical parameters. The expression of EBI3 and Foxp3 mRNA in peripheral blood mononuclear cells of patients with sarcoidosis (1.54±0.74, 0.92±0.36) were significantly lower than those in the normal control group respectively (2.12±0.61, 1.10±0.27, all P<0.05). There was a significant positive correlation between the expression of EBI3 and Foxp3 mRNA in the sarcoidosis group (r=0.786, P<0.001). Conclusion: IL-35 may be involved in the inflammatory process of sarcoidosis and play an important role in the pathogenesis of the disease.

[Precision medical treatment of non-adenocarcinoma NSCLC patients with EGFR mutation].

EGFR tyrosine kinase inhibitors (TKIs) treatment has been established as standard therapy for EGFR-mutated adenocarcinomas. In the studies which published prospective randomized trials comparing EGFR TKIs with chemotherapy, a very low percentage of EGFR-mutated non-adenocarcinomas was enrolled in clinical trials. The efficacy of TKIs treatment for EGFR-mutated non-adenocarcinomas and their relationship with clinicopathological characteristics remain debatable. The results of retrospective studies show that the frequency of EGFR mutation is lower in non-adenocarcinoma than that of adenocarcinoma and efficacy of TKIs treatment for non-adenocarcinoma is inferior to adenocarcinoma. Smoking status is significantly associated with the efficacy of TKIs treatment for EGFR-mutated non-adenocarcinomas. The EGFR mutation rate and efficacy of TKIs treatment in adenosquamous cell carcinoma are higher than those of squamous cell carcinoma or large cell lung carcinoma. It may be concluded that the incidence of EGFR mutations in patients with non-adenocarcinoma NSCLC from mainland China is not very low and it is reasonable that EGFR TKIs could be an option for the treatment of EGFR-mutated non-adenocarcinoma NSCLC, especially for patients with adenosquamous histology and non-smokers. It is necessary to conduct a large-sample prospective study to understand the clinicopathological characteristics of non-adenocarcinomas and to evaluate the efficacy of EGFR TKI treatment or/and chemotherapy for EGFR-mutated non-adenocarcinoma NSCLC.

[Cancer-associated-fibroblasts regulate the chemoresistance of lung cancer cell line A549 via SDF-1 secretion].

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05). Conclusions: Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

[Coexistence of sarcoidosis and primary Sjögren syndrome: a clinical analysis and literature review].

Four patients with coexistence of sarcoidosis and primary Sjögren syndrome (pSS) were retrospectively analyzed.All patients were female, who were referred to our department mainly because of respiratory symptoms.Positive antinuclear antibody(ANA) was detected in 2 patients and anti-Sjögrens syndrome A (SSA) antibody positive in 1 patient.All patients presented specific histologic patterns of both sarcoidosis and pSS.Publications related to coexistence of these two diseases were reviewed.Forty-one patients were finally included in the analysis, among whom 37 confirmed patients were from literature search.There were 37 women and 4 men.The main clinical features presentation were xerophthalmia in 40, xerostomia in 38, hilaradenopathies in 28, interstitial lung disease in 15, respiratory symptoms in 13.The main immunologic data were positive ANA in 23, SSA antibody in 19, anti-Sjögrens syndrome B antibody in 10 and rheumatoid factor in 12.All patients presented specific histologic patterns of both diseases.Patients with both sarcoidosis and pSS of ten represent multisystemic involvement and positive immunologic parameters, as well as the dual expression of specific histologic characteristics.

[Clinical and radiological features of adult patients with different antisynthetase syndrome based on serum anti-aminoacyl-tRNA synthetase antibody type].

Objective: To describe the clinical and radiological characteristics of antisynthetase syndrome associated interstitial lung disease in patients with different serum anti-aminoacyl-tRNA synthetase antibodies. Methods: We conducted a retrospective analysis of 5 adult patients with antisynthetase syndrome associated interstitial lung disease in Peking Union Medical College Hospital. Their clinical and chest radiological data were analyzed and relevant literatures were reviewed. Results: Among these 5 patients, there were 1 male and 4 females, aged from 32 y to 67 y, with a mean age of 53 y. Cough and exertional dyspnea were the main clinical complaints. Four cases showed mechanic's-like hands, and all of the 5 cases had Velcro rales in the basal lungs. None of them showed clubbing. Creatine kinase was elevated in 1 case with anti-Jo-1 synthetase antibody, and anti-nuclear antibody was positive in 4 cases, with different titers from 1∶80 to 1∶320, and anti-Ro-52 antibody was positive in 3 cases. Anti-aminoacyl-tRNA synthetase antibody spectrum analysis showed 1 case with anti-Jo-1, 1 anti-PL-7, 1 anti PL-12, 1 anti-EJ and 1 anti-OJ synthetase antibody, respectively. Chest high resolution CT showed nonspecific interstitial pneumonia pattern in 1 case, and nonspecific interstitial pneumonia pattern with organizing pneumonia pattern in 4 cases. All the cases responded to immunosuppressive therapy including corticosteroids (with starting prednisone dosage more than 1 mg·kg(-1)·d(-1)) plus azathioprine or Mycophenolate mofetil. Conclusions: Antisynthetase syndrome associated interstitial lung disease, characterized by the presence of different anti-tRNA synthetase antibodies, is an increasingly recognized clinical entity. Clinical and radiological features of different subtypes of antisynthetase syndrome are relatively heterogeneous. Nonspecific interstitial pneumonia pattern, nonspecific interstitial pneumonia pattern with organizing pneumonia pattern and organizing pneumonia pattern were common chest HRCT patterns. Prednisone plus immunosuppressive agents are the recommended first line therapy.

Genome-wide analysis of the maize (Zea may L.) CPP-like gene family and expression profiling under abiotic stress.

Cysteine-rich polycomb-like (CPP) proteins are members of a small family of transcription factors, which have been identified and characterized in Arabidopsis, rice, and soybean. In this study, we investigated CPP-like genes in the maize genome. The results revealed 13 putative CPP-like genes, which were found to encode 17 distinct transcripts and were distributed unequally on 7 of 10 maize chromosomes. Analysis of phylogenetic relationships showed that Arabidopsis, rice, and maize CPP-like transcription factors can be grouped into two subfamilies. We also used real-time RT-PCR to evaluate changes in the transcript levels of ZmCPP genes in response to abiotic stresses (heat, cold, salt, and drought stresses). These findings provide an overview of the evolution of the ZmCPP gene family, which will aid in the functional characterization of CPP-like genes in maize growth and development.

Magnetoinfrared Spectroscopy of Landau Levels and Zeeman Splitting of Three-Dimensional Massless Dirac Fermions in ZrTe(5).

We present a magnetoinfrared spectroscopy study on a newly identified three-dimensional (3D) Dirac semimetal ZrTe(5). We observe clear transitions between Landau levels and their further splitting under a magnetic field. Both the sequence of transitions and their field dependence follow quantitatively the relation expected for 3D massless Dirac fermions. The measurement also reveals an exceptionally low magnetic field needed to drive the compound into its quantum limit, demonstrating that ZrTe(5) is an extremely clean system and ideal platform for studying 3D Dirac fermions. The splitting of the Landau levels provides direct, bulk spectroscopic evidence that a relatively weak magnetic field can produce a sizable Zeeman effect on the 3D Dirac fermions, which lifts the spin degeneracy of Landau levels. Our analysis indicates that the compound evolves from a Dirac semimetal into a topological line-node semimetal under the current magnetic field configuration.

Status and future of Quantitative Microbiological Risk Assessment in China.

Since the implementation of the Food Safety Law of the People's Republic of China in 2009 use of Quantitative Microbiological Risk Assessment (QMRA) has increased. QMRA is used to assess the risk posed to consumers by pathogenic bacteria which cause the majority of foodborne outbreaks in China. This review analyses the progress of QMRA research in China from 2000 to 2013 and discusses 3 possible improvements for the future. These improvements include planning and scoping to initiate QMRA, effectiveness of microbial risk assessment utility for risk management decision making, and application of QMRA to establish appropriate Food Safety Objectives.

Associations between clinical characteristics and oncogene expression in patients with non-small cell lung cancer.

More than 40 oncogenes associated with non-small cell lung cancer (NSCLC) have been identified with varied gene expression. The correlations between specific clinical characteristics and oncogene expression in NSCLC patients were examined. From October 2011 to September 2012, a total of 60 patients with NSCLC (male:female, 34:24; mean age, 59.5 ± 10.6 years; age range, 31-81 years) were diagnosed and evaluated for treatment with radical resection at a single facility. Eligible patients exhibiting tumor node metastasis (TNM) stage I-III NSCLC confirmed by post-surgical pathology were included. mRNA expression was detected by branched DNA-liquidchip technology (bDNA-LCT) and mutations were detected at EGFR exons 18, 19, 20, and 21, KRAS exons 2 and 3, BRAF and PIK3CA exons 9 and 20. Correlations between gene expression at mutations and clinical characteristics of gender, age, histological type, degree of differentiation, smoking status, immunohistochemical (IHC) evaluation of TTF-1, TNM staging, and discrete age ("nage") were examined. Significant associations were observed between IHC staining for TTF-1 and histological type (P = 0.00001) and with BRAC1, TYMS, RRM1, and TUBB3 expression (P = 0.0187, 0.0051, 0.024, and 0.0238, respectively). Significant cross-correlations were observed between TYMS, BRAC1, TOP2A, STMN1, TUBB3, and RRM1 expression (P < 0.05), but not between EGFR exon 21, KRAS exon 2, and PIK3CA exon 9 expression and any other mutation expression (P > 0.05). Relationships between clinical characteristics and oncogene expression in NSCLC, particularly those of TTF-1 level and smoking status, may be useful indicators of prognosis and development of anti-cancer drug resistance.

Secretory leukocyte protease inhibitor mediates non-redundant functions necessary for normal wound healing.

Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor with anti-microbial properties found in mucosal fluids. It is expressed during cutaneous wound healing. Impaired healing states are characterized by excessive proteolysis and often bacterial infection, leading to the hypothesis that SLPI may have a role in this process. We have generated mice null for the gene encoding SLPI (Slpi), which show impaired cutaneous wound healing with increased inflammation and elastase activity. The altered inflammatory profile involves enhanced activation of local TGF-beta in Slpi-null mice. We propose that SLPI is a pivotal endogenous factor necessary for optimal wound healing.

[Clinical genetic analysis and diagnosis of a family with hereditary hemorrhagic telangiectasia].

Objective: To explore the diagnostic significance of the combination of clinical and genetic detection of hereditary hemorrhagic telangiectasia (HHT) by analyzing the clinical and genetic diagnosis of a family with HHT. Methods: Medical history data of the probands and their family members were collected, and the sequence analyses of coding regions of ENG, ACVRL1, SMAD4 and GDF2 genes were performed by PCR-sequencing method, and a comprehensive diagnosis was made based on the clinical features and gene detection results. After the pathogenic gene variation was identified, 11 members of 3 generations of the family were tested for pathogenic gene mutation. Results: There was an ACVRL1 c.715_716delAG mutation in the proband and 9 other family members, which caused p.S239C. Based on the clinical and genetic findings, the 7 suspected were diagnosed and 2 asymptomatic patients were found to carry the mutation site. Conclusion: The combination of clinical features and gene detection can determine the etiology and classification of HHT, which is convenient for the early diagnosis and prevention of the disease.

[Self-rated health measures and their relations to all-cause and cardiovascular mortality in adults from 10 regions of China].

Objective: To examine the association between self-rated health status (SRH) and all-cause and cardiovascular mortality. Methods: A total of 512 713 adults aged 30-79 years from 10 areas of China were followed from baseline (2004-2008) until 31 December 2016 in the China Kadoorie Biobank study. Global and age-comparative SRH [general self-rated health status (GSRH) and age-comparative self-rated health status (ASRH), respectively] were asked in baseline questionnaires. Causes for mortality were monitored through linkage with established Disease Surveillance Point system and health insurance records. Multivariable Cox proportional regression models were used to estimate the HRs and 95%CIs for the association between SRH measures and all-cause or cardiovascular mortality. Results: During an average of 9.9 years' follow-up, 44 065 deaths were recorded, among which 17 648 were from cardiovascular disease. Compared with excellent GSRH, the HR(95%CI) for all-cause and cardiovascular mortality associated with poor GSRH was 1.84(1.78-1.91) and 1.94(1.82-2.06), respectively. Relative to better ASRH, the HR(95%CI) for all-cause and cardiovascular mortality associated with worse ASRH was 1.75(1.70-1.81) and 1.83(1.73-1.92), respectively. Conclusion: In this large prospective cohort study in China, participants reporting poor GSRH or worse ASRH had significantly higher risk of all-cause and cardiovascular mortality.

Dietary supplementation of methionine mitigates oxidative stress in broilers under high stocking density.

We aimed to investigate whether dietary supplementation of methionine could mitigate intestinal oxidative injury in broilers under high stocking density (HSD). In the grower phase (d 22-42), 576 broilers with similar body weight were randomly chosen and divided into 8 groups in a 2 × 4 factorial experiment. Two different stocking densities (14 and 20 broilers per m2) were tested with 4 different methionine levels: 0.35%, 0.4%, 0.45%, or 0.5%. Intestinal morphological and oxidative stress markers were assessed at the end of the test period. The results showed that mortality of broilers was significantly higher in the HSD group fed 0.35% methionine diet than the other groups, which was reversed by supplementation with 0.40% to 0.50% methionine. HSD significantly decreased feed intake and daily weight gain. HSD treatment significantly decreased T-AOC, activity of GPX (P < 0.01) and increased the level of PCO (P < 0.01), MDA (P = 0.052) of plasma. The decreased glutathione peroxidase activity in the liver and jejunum caused by HSD was alleviated by additional methionine. Supplementation of methionine increased the ration of GSH/GSSG in the plasma. The jejunum villus height and ratio of villus height to crypt depth under low stocking density conditions with 0.40% methionine diet were the highest, whereas the 0.45% methionine group was the highest under HSD conditions. Thus, additional dietary supplementation of methionine mitigates oxidative stress in broilers under HSD conditions and 0.40% to 0.45% methionine can be applied in cage rearing broiler production for amelioration of oxidative stress caused by HSD.