[The Breast Cancer Cohort Study in Chinese Women: research design and preliminary results of clinical multi-center cohort].

Objective: The incidence of breast cancer in Chinese women continues to rise. The large breast cancer cohort studies in China are relatively scarce. There are many bottlenecks in the construction of large clinical cohort for breast cancer diagnosis, treatment, and prognoses, such as inconsistent standards, high rates of lost follow-up, repeated construction, and inability to share. To better solving the difficulties and problems faced by large-scale clinical cohort research in China, this project will cooperate with several tertiary A hospitals to establish a breast cancer cohort in Chinese women. It also provides a data platform and technical support for breast cancer multi-center clinical cohort research. Methods: Based on the evidence-based medicine and expert opinion and consensus, we established a breast cancer cohort standardized indicator set-recording baseline information, diagnosis and treatment-related information of the enrolled patients, and collecting biological specimens. According to the technical specification of long-term follow-up for the endpoint, data management, and data security and in the large population-based cohort study, a standardized follow-up system for the diagnosis, treatment, and prognosis of breast cancer prospective cohorts is formed. Results: Based on standardized data sets and the computer discipline's advantage from the University of Science and Technology Beijing, we integrate the new information technology methods, including dynamic information collection terminals and social networks. Thus, the quality of control programs on compliance and intelligence data was improved, and a Chinese women breast cancer cohort database was developed. By February 2020, 12 147 patients were included in the clinical cohort database. Biological specimens'resources in cohort construction were collected and cooperated with Shandong University to research the multi-center quality control system and shared evaluation system of biobanks. Building an open and shared biobank network and forming a full chain of breast cancer research platform. Conclusion: With the implementation of the "13(th) Five-Year Plan" precision medicine research, this study provides a research foundation for precision diagnosis and treatment of breast cancer and provides data support for the country to formulate relevant medical policies.

Determination of the potential of induced pluripotent stem cells to differentiate into mouse nucleus pulposus cells in vitro.

We determined the potential for induced pluripotent stem (iPS) cells to differentiate into nucleus pulposus (NP)-like cells in mice. iPS cells were generated from tail-tip fibroblasts. We used a pellet culture model with the aim of determining the applicability of iPS cell-based therapy to intervertebral disc degeneration (IVD). The cell pellet was cultured in an NP cell basal medium comprising Dulbecco's modified Eagle's medium supplemented with transforming growth factor beta 1, dexamethasone, ascorbate-2-phosphate, and 1% ITS-Premix. The pellet was evaluated by quantitative reverse transcription polymerase chain reaction, immunohistochemical staining, and biochemical composition. The differentiation of iPS cells into NP cells was demonstrated by the protein and mRNA expression levels of proteoglycan, collagen II, aggrecan, and CD24. Furthermore, increased hydroxyproline content and dimethylmethylene blue staining demonstrated that the collagen II and glycosaminoglycan content in the NP cells increased with time. We have shown that cultured mouse iPS cells can be induced to differentiate into NP cells. Such proof-of-concept opens up the possibility of producing patient-specific NP cells in a relatively simple and straightforward manner with high efficiency. We are confident that such cells could be immediately useful for the study of IVD disease.

Analysis of the differentially expressed genes in Microsporum canis in inducing smooth skin and scalp tissue conditions.

Microsporum canis is a common zoophilic dermatophyte, which causes a range of infections. To explore the pathogenic mechanism of tinea capitis, we used the suppression subtractive hybridization (SSH) technique to investigate the differences in gene expression between different cultures of Microsporum canis incubated on three different types of mineral media containing child glabrous skin, child scalp tissue and adult scalp tissue. Using dot-blot hybridization and real-time PCR technique, we successfully screened and identified a pair of genes that had expression levels 44.6 and 117 times higher in culture 1 (M. canis cultured in mineral medium with child scalp tissue) than in culture 2 (M. canis cultured in mineral medium with glabrous skin tissue), and another pair of genes with expression levels 78.2 and 9.8 times higher in culture 1 than in culture 3 (M. canis cultured in mineral medium with adult scalp tissue). These four genes were found to have 41%, 53%, 40% and 94% homology to those encoding a hypothetical protein [family of serine hydrolases 1; (FSH1)], PQ loop repeat protein (PQ-LRP), a predicted protein [porphyrin galactose 4; (P-GAL4)] and NADH dehydrogenase subunit (NADH)1, respectively. The upregulation of the FSH1, PQ-LRP, P-GAL4 and NADH1 genes in cultures of child scalp tissue indicates that they are essential in the pathogenesis of tinea capitis caused by M. canis.

Introduction of a disulfide bridge enhances the thermostability of a Streptomyces olivaceoviridis xylanase mutant.

Substitution of the N-terminus of Streptomyces olivaceoviridis xylanase XYNB to generate mutant TB has been previously shown to increase the thermostability of the enzyme. To further improve the stability of this mutant, we introduced a disulfide bridge (C109-C153) into the TB mutant, generating TS. To assess the effect of the disulfide bridge in the wild-type enzyme, the S109C-N153C mutation was also introduced into XYNB, resulting in XS. The mutants were expressed in Pichia pastoris, the recombinant enzymes were purified, and the effect of temperature and pH on enzymatic activity was characterized. Introduction of the disulfide bridge (C109-C153) into XYNB (XS variant) and TB (TS variant) increased the thermostability up to 2.8-fold and 12.4-fold, respectively, relative to XYNB, after incubation at 70 degrees C, pH 6.0, for 20 min. In addition, a synergistic effect of the disulfide bridge and the N-terminus replacement was observed, which extended the half-life of XYNB from 3 to 150 min. Moreover, XS and TS displayed better resistance to acidic conditions compared with the respective enzymes that did not contain a disulfide bridge.