Proteomics study of Mycoplasma pneumoniae pneumonia reveals the Fc fragment of the IgG-binding protein as a serum biomarker and implicates potential therapeutic targets / 医学前沿

Macrolide and corticosteroid resistance has been reported in patients with Mycoplasma pneumoniae (MP) pneumonia (MPP). MP clearance is difficult to achieve through antibiotic treatment in sensitive patients with severe MPP (SMPP). SMPP in children might progress to airway remodeling and even bronchiolitis/bronchitis obliterans. Therefore, identifying serum biomarkers that indicate MPP progression and exploring new targeted drugs for SMPP treatment require urgency. In this study, serum samples were collected from patients with general MPP (GMPP) and SMPP to conduct proteomics profiling. The Fc fragment of the IgG-binding protein (FCGBP) was identified as the most promising indicator of SMPP. Biological enrichment analysis indicated uncontrolled inflammation in SMPP. ELISA results proved that the FCGBP level in patients with SMPP was substantially higher than that in patients with GMPP. Furthermore, the FCGBP levels showed a decreasing trend in patients with GMPP but the opposite trend in patients with SMPP during disease progression. Connectivity map analyses identified 25 possible targeted drugs for SMPP treatment. Among them, a mechanistic target of rapamycin kinase (mTOR) inhibitor, which is a macrolide compound and a cell proliferation inhibitor, was the most promising candidate for targeting SMPP. To our knowledge, this study was the first proteomics-based characterization of patients with SMPP and GMPP.

Post-surgical resection prognostic value of combined OPN, MMP7, and PSG9 plasma biomarkers in hepatocellular carcinoma / 医学前沿

Biomarkers for hepatocellular carcinoma (HCC) following curative resection are not currently sufficient for prognostic indication of overall survival (OS) and disease-free survival (DFS). The aim of this study was to investigate the prognostic performance of osteopontin (OPN), matrix metalloproteinase 7 (MMP7), and pregnancy specific glycoprotein 9 (PSG9) in patients with HCC. A total of 179 prospective patients with HCC provided plasma before hepatectomy. Plasma OPN, MMP7, and PSG9 levels were determined by enzyme-linked immunosorbent assay. Correlations between plasma levels, clinical parameters, and outcomes (OS and DFS) were overall analyzed. High OPN ( ⩾ 149.97 ng/mL), MMP7 ( ⩾ 2.28 ng/mL), and PSG9 ( ⩾ 45.59 ng/mL) were prognostic indicators of reduced OS (P < 0.001, P < 0.001, and P = 0.007, respectively). Plasma PSG9 protein level was an independent factor in predicting OS (P = 0.008) and DFS (P = 0.038). Plasma OPN + MMP7 + PSG9 elevation in combination was a prognostic factor for OS (P < 0.001). OPN was demonstrated to be a risk factorassociated OS in stage I patients with HCC and patients with low α-fetoprotein levels ( < 20 ng/mL). These findings suggested that OPN, MMP7, PSG9 and their combined panels may be useful for aiding in tumor recurrence and mortality risk prediction of patients with HCC, particularly in the early stage of HCC carcinogenesis.

Characterization of Distinct T Cell Receptor Repertoires in Tumor and Distant Non-tumor Tissues from Lung Cancer Patients / 基因组蛋白质组与生物信息学报·英文版

T cells and T cell receptors (TCRs) play pivotal roles in adaptive immune responses against tumors. The development of next-generation sequencing technologies has enabled the analysis of the TCRβ repertoire usage. Given the scarce investigations on the TCR repertoire in lung cancer tissues, in this study, we analyzed TCRβ repertoires in lung cancer tissues and the matched distant non-tumor lung tissues (normal lung tissues) from 15 lung cancer patients. Based on our results, the general distribution of T cell clones was similar between cancer tissues and normal lung tissues; however, the proportion of highly expanded clones was significantly higher in normal lung tissues than in cancer tissues (0.021% ± 0.002% vs. 0.016% ± 0.001%, P = 0.0054, Wilcoxon signed rank test). In addition, a significantly higher TCR diversity was observed in cancer tissues than in normal lung tissues (431.37 ± 305.96 vs. 166.20 ± 101.58, P = 0.0075, Mann-Whitney U test). Moreover, younger patients had a significantly higher TCR diversity than older patients (640.7 ± 295.3 vs. 291.8 ± 233.6, P = 0.036, Mann-Whitney U test), and the higher TCR diversity in tumors was significantly associated with worse cancer outcomes. Thus, we provided a comprehensive comparison of the TCR repertoires between cancer tissues and matched normal lung tissues and demonstrated the presence of distinct T cell immune microenvironments in lung cancer patients.

In vitro method for skin sensitization based on dendritic cell activation / 中国比较医学杂志

Allergy contact dermatitis is a type IV delayed type hypersensitivity that is induced by exogenous compounds and involves many cell types. Traditional animal testing uses guinea pigs or mice as a model. With progress of the adverse outcome pathway(AOP)on skin sensitization, a concept for development of alternative methods based on a molecular initiating event and key events is provided. Dendritic cell(DC)activation plays a key role in the AOP. Many alternative methods have been developed,with several methods validated and accepted as guidance for assays. This paper examines DC screening, characteristics of test parameters, and limitations and applicability of DC-derived methods. Progress on interactions between DCs and other cells, co-culture systems, and the human body-on-a-chip will also be introduced. Altogether,this paper will provide information for optimization of in vitro alternative methods for sensitization detection.

An in vitro skin sensitization test based on THP-1 cell line / 中国比较医学杂志

Objective To establish an in vitro skin sensitization test,human cell line activation test (h-CLAT),based on THP-1 cell line (a human acute monocytic leukemia cell line),and to assess the sensitizing potency of plant raw materials of chemical and cosmetic products by this in vitro skin sensitization test.Method THP-1 cells were cultured in vitro and exposed to 11 reference skin sensitization chemicals and 9 samples,by monitoring the cell viability,cell surface marker CD54 /CD86 and relative fluorescence intensity of cells surface after the cells was exposures to the substances,and to discover whether there is a positive reaction.At the same time,Buehler test was used to validate the results of samples tested by h-CLAT.Results 11 reference chemicals were distinguished correctly by h-CLAT.Among the 9 samples tested,7 samples were recognized as negative sensitizer and 2 plant extracted substances were identified as suspicious skin sensitizer.The qualitative classification of the 9 samples by h-CLAT test was consistent with the results obtained by animal test.Conclusions The h-CLAT-in vitro test can be used to replace some animal tests for the prediction of soluble skin sensitizing substances.

Preliminary study for integrating DPRA with h-CLAT to predict skin sensitizers / 中国实验动物学报

Objective To establish a detection method integrating DPRA ( direct peptide reactivity assay) with h?CLAT ( human cell line activation test) to screen the skin sensitization potency of chemicals and plant extracts. Methods 12 chemicals and 7 plant extracts were chosen as the test substances. Firstly, the test substances were incubated together with two different peptides ( cysteine and lysine) respectively for reaction for 24 h. The peptide consumptions were analyzed by HPLC. Simultaneously, THP?1 cells were cultured in vitro and then exposed to different concentrations of test sub?stances for 24 h to examine the cell viability, cell surface markers CD54 and CD86 were assessed by flow cytometry. The predicting results were compared further between DPRA and h?CLAT. Results 12 chemicals were distinguished correctly by DPRA classified as 2 non?sensitizers and 10 sensitizers. The results of DPRA were in accordance with h?CLAT. Predic?ting the sensitization potency of plant extracts by DPRA showed that 6 plant extracts were determined as suspected sensiti?zers except for green tea extract. But using the method of h?CLAT, 4 plant extracts were examined as suspected sensitizers except for green tea extract, herba portulacae extract and ginseng fruit extract. The coherence of DPRA and h?CLAT was 0?57. Conclusion This detection method integrating DPRA with h?CLAT can predict single compound accurately. As for complex compound, it can achieve preliminary prediction and need other integrating methods to make a further identifica?tion.

OLC1 protein levels in plasma of patients with non-small cell lung cancer and its clinical application / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to detect the plasma concentration of OLC1 (overexpressed in lung cancer 1) protein as a potential cancer biomarker, and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>We prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice. Each mouse was immunized four times at a dose of 15-30 µg antigen protein, and the interval between two consecutive immunizations was two weeks. Antibody screening was made by ELISA and Western blot, and a double antibody sandwich ELISA kit was developed. We used this established ELISA kit to detect the plasma concentration of OLC1 protein in 281 NSCLC patients and 92 gender- and age-matched healthy controls. Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC1.</p><p><b>RESULTS</b>We obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml. OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P < 0.001). In the scenario of distinguishing NSCLC from control group, AUC result was 0.69. When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4% and 51.1%, respectively. In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively.</p><p><b>CONCLUSION</b>The plasma concentration of OLC1 protein is significantly elevated in NSCLC patients. OLC1 may be as a potential cancer biomarker applied in clinical diagnosis.</p>

Integrated CAMVA and BCOP methods to predict eye irritation caused by cosmetics / 中国比较医学杂志

Objective To explore the use of integrated two methods in vitro in prediction of eye irritation caused by cosmetics.Method Chorioallantoic membrane vascular assay ( CAMVA), bovine corneal opacity and permeability (BCOP) and Draize rabbit eye irritation test were used to determine the predictive potential of eye irritation of 60 kinds of cosmetics.Results CAMVA method was able to distinguish 41 non-irritant samples and 18 irritant samples.BCOP method was able to predict 35 non-irritant samples , 21 mild-moderate irritant samples and 4 severe irritant samples . Combination of CAMVA and BCOP methods could obviously improve the identification ability of irritation , and the classification consistency with Draize rabbit eye irritation testing reached 98.3%.Conclusions The integrated test strategy combined BCOP with CAMVA can be used to appropriately predict ocular irritation of cosmetics , with a prediction range covering non-irritant to severe irritant samples .

Relation between apoptosis and proliferation or genes expression after chemotherapy in ovarian eithelial carcinoma / 中国癌症杂志

Purpose:To study the apoptosis and proliferation in ovarian cancer and the relationship between gene and the histological grading as well as response to chemotherapy. Methods:In our study apoptosis in fifty fresh samples of epithelial ovarian carcinomas were evaluated by TDT (terminal deoxynucleotidyl transferase) assay. We used the techniques of immunohistochemistry to detect the proliferation and the protein expression of bcl-2,bax and p53 gene.Results:In 66% (30/50) samples of fresh epithelial ovarian carcinoma apoptosis was observed in different degrees. Our results showed that PCAN and p53 expression in poorly differentiated ovarian carcinoma is higher than that in well middle differentiated (P

Alteration of apoptotic susceptibility and bcl-2 gene in malignant transformation of human bronchial epithelial cells / 中国病理生理杂志

AIM: To demonstrate the susceptibility of cell apoptosis varies during the progress of cell malig- nant transformation from human being in vitro. METHODS: A SV40T - transfected human bronchial epithelial im- mortalized cell line (called M) was selected in this work, which has acquired some characteristics of malignant trans- formation at the later passage. The alterations of apoptosis and bcl- 2, P53 genes between early and later passage of M cells were investigated by means of TDT labeling in situ, chromosome FISH, RNA and protein testing, etc. RE- SULTS: Incidence of apoptosis induced by cis - platin was significantly lower in later than in early passages of M. Levels of bcl - 2 mRNA and protein in later passages were higher than early passages of M, and overxpression of bcl -2 was accumulated following the development of cellular malignancy. P53 protein level was as high in early as in later passages. CONCLUSION: Overexpression of bcl - 2 decreases the cellular sensitivity to apoptotic inductors plays an important role during progress of carcinogenesis in human bronchial epithelial cancers. The inactivation of P53 protein in the SV40 - T transfected M cell line may be one of reasons of bcl - 2 overexpression, but not associated with the accumulation of bcl - 2 expressed level during cell transformation.

RESEARCH ON THE COMMITTED DIFFERENTIATION OF EMBRYONIC STEM CELLS INTO EPIDERMAL-LIKE STEM CELLS INDUCED in vitro / 解剖学报

Objective To investigate the condition which can induce mouse ES cells to differentiate into epidermal-like stem cells for the clinical application of ES cells-derived epidermal-like stem cells and the research on the mechanism of committed differentiation of ES cells. Methods Coculture mouse ES cells with human amnion for 3-4 days, and the committed differentiation were detected by flow cytometry and immunohistochemistry. In experimental group 1, amnion was pasted covering the whole bottom of the wells, with the epithelial surface upward, and in group 2 covering the half bottom. No amnion was used in control. Results After 3-4 days of co-culture, epidermal-like stem cell clones were formed on the epithelial surface of amnion in group 1 and group 2, and expressed high levels of integrin-? 1, CK19 and CK15. The percentages of integrin-? 1, CK19 and CK15 positive cells counted in group 1 by flow cytometry were 89.2%, 86.8% and 71.2% respectively, versus the control group of 8.4%,9.6% and 11.8%, the differences were significant in all the three indices (Z tests P