Integrative analysis of differentially expressed mRNAs and proteins induced by PGC-1ß in breast cancer cells.

Breast cancer is one of the most frequent malignancies in females. The molecular mechanism of how breast cancer development and recurrence still need to be explored. Peroxisome gamma coactivator-1ß (PGC-1ß) was engaged in cancer energy metabolism and tumor genesis. However, the mechanisms of PGC-1ß in breast cancer have not been fully understood. In this study, PCG-1ß overexpressed and knockdown vectors were transferred into MCF-7 cells. With the association-quantitative connection analysis, the different expressions of mRNAs and proteins were examined. Additionally, the terms on differentially expressed mRNAs and proteins were enriched by GO and KEGG. Based on the results, 1872 differentially expressed genes were identified in the up-regulated of PGC-1ß group, and 1318 genes were found in the down-regulated of PGC-1ß cells. With the label-free technique, 221 differentially expressed proteins were screened in PGC-1ß up-regulated group, and 459 proteins were identified in PGC-1ß down-regulated group. Correlation analysis showed that 49 significantly expressed mRNA-protein pairs in OV vs CT groups and 25 paired in SI vs CT groups. Combined analysis of transcriptome and proteome demonstrated that PGC-1ß plays a important role in cancer energy metabolism and boosting the pace of chemical processes in the proliferation of breast cancer cells. Additional investigation about PGC-1ß and energy metabolism in cancer cells may shed fresh light on the growth and treatment of breast cancer cells.

An entropy-driven three-dimensional multipedal-DNA walker for ultrasensitive detection of cancer cells.

Sensitive and accurate detection of cancer cells is of great significance for the early diagnosis and treatment of cancer. In this work, we developed a simple fluorescent signal amplification biosensor based on an entropy-driven three-dimensional (3D) multipedal-DNA walker for highly sensitive detection of cancer cells. Firstly, DNA tetrahedron nanostructures (DTNs) combined with AS1411 aptamer were used as the capture probe to achieve efficient capture of cancer cells. Then, the bipedal hairpin fuel chain hybridized with DTNs and exposed two catalytic "legs" to form a walker probe. Finally, the walker probe autonomously walked on polystyrene microspheres (PS) via entropy-driven catalytic reaction. DTNs rolled on the PS to achieve multipedal walking, realizing fluorescence signal amplification due to fluorescence recovery of DNA-CdTe quantum dots on the PS surface. This fluorescence signal amplification strategy showed excellent selectivity and sensitivity toward cancer cells with the detection limit of 7 cell mL-1. This entropy-driven 3D multipedal DNA walker fluorescence exhibited great potential in detecting circulating tumor cells and tumor markers used for early diagnosis and clinical treatment of cancer.

A sustainable mouse karyotype created by programmed chromosome fusion.

Chromosome engineering has been attempted successfully in yeast but remains challenging in higher eukaryotes, including mammals. Here, we report programmed chromosome ligation in mice that resulted in the creation of new karyotypes in the lab. Using haploid embryonic stem cells and gene editing, we fused the two largest mouse chromosomes, chromosomes 1 and 2, and two medium-size chromosomes, chromosomes 4 and 5. Chromatin conformation and stem cell differentiation were minimally affected. However, karyotypes carrying fused chromosomes 1 and 2 resulted in arrested mitosis, polyploidization, and embryonic lethality, whereas a smaller fused chromosome composed of chromosomes 4 and 5 was able to be passed on to homozygous offspring. Our results suggest the feasibility of chromosome-level engineering in mammals.

Aptamer-mediated DNA concatemer functionalized magnetic nanoparticles for reversible capture and release of circulating tumor cells.

Effectively capturing, releasing, and reanalyzing circulating tumor cells (CTCs) are critical in cancer diagnosis and individualized treatment. Traditional immunomagnetic separation has disadvantages of low sensitivity and specificity, and is time-consuming and costly in CTCs capture. It is also easily disturbed by the microenvironment in releasing and analyzing CTCs. Here, we proposed an aptamer-mediated DNA concatemer functionalized magnetic nanoparticles (MNPs-AMDC) for the reversible capture and release of CTCs. In this study, aptamers were used both for efficiently capturing CTCs without complicated assembly steps and stimulus-response switch for releasing CTCs with little influence on cellular activity. The MNPs-AMDC was demonstrated to effectively capture (83%) and release CTCs with a good viability rate (92%). Moreover, this device was also tested in clinical blood samples, which would provide a universal tool for diagnosing cancer and treating individuals.

LncSNHG1 Promoted CRC Proliferation through the miR-181b-5p/SMAD2 Axis.

Objective: To investigate the effects of LncRNA SNHG1 on the proliferation, migration, and epithelial-mesenchymal transition (EMT) of colorectal cancer cells (CRCs). Methods: 4 pairs of CRC tissue samples and their corresponding adjacent samples were analyzed by the human LncRNA microarray chip. The expression of LncSNHG1 in CRC cell lines was verified by qRT-PCR. Colony formation assays and CCK8 assays were applied to study the changes in cell proliferation. The transwell assay and wound healing experiments were used to verify the cell invasion and migration. EMT progression was confirmed finally. Results: LncSNHG1 was overexpressed both in CRC tissues and cell lines, while the miR-181b-5p expression was decreased in CRC cell lines. After knock-down of LncSNHG1, the proliferation, invasion, and migration of HT29 and SW620 cells were all decreased. Meanwhile, LncSNHG1 enhanced EMT progress through regulation of the miR-181b-5p/SMAD2 axis. Conclusion: LncSNHG1 promotes colorectal cancer cell proliferation and invasion through the miR-181b-5p/SMAD2 axis.

The Role of Non-Coding RNAs in Breast Cancer Drug Resistance.

Breast cancer (BC) is one of the commonly occurring malignancies in females worldwide. Despite significant advances in therapeutics, the mortality and morbidity of BC still lead to low survival and poor prognosis due to the drug resistance. There are certain chemotherapeutic, endocrine, and target medicines often used for BC patients, including anthracyclines, taxanes, docetaxel, cisplatin, and fluorouracil. The drug resistance mechanisms of these medicines are complicated and have not been fully elucidated. It was reported that non-coding RNAs (ncRNAs), such as micro RNAs (miRNA), long-chain non-coding RNAs (lncRNAs), and circular RNAs (circRNAs) performed key roles in regulating tumor development and mediating therapy resistance. However, the mechanism of these ncRNAs in BC chemotherapeutic, endocrine, and targeted drug resistance was different. This review aims to reveal the mechanism and potential functions of ncRNAs in BC drug resistance and to highlight the ncRNAs as a novel target for achieving improved treatment outcomes for BC patients.

Mortalin maintains breast cancer stem cells stemness via activation of Wnt/GSK3ß/ß-catenin signaling pathway.

Previous research indicated that mortalin overexpressed in breast cancer and contributed to carcinogenesis. Mortalin was also demonstrated to promote Epithelial-mesenchymal transition (EMT) and was considered as a factor for maintaining the stemness of the cancer stem cells. However, the underlying mechanisms about mortalin maintaining the stemness of breast cancer stem cells (BCSCs) remain unclear. Here, we identified that increased expression of mortalin in breast cancer was associated with poorer overall survival rate. Mortalin was elevated in breast cancer cell lines and BCSC-enriched populations. Additionally, knockdown of mortalin significantly inhibited the cell proliferation, migration and EMT, as well as sphere forming capacity and stemness genes expression. Further study revealed that mortalin promoted EMT and maintained BCSCs stemness via activating the Wnt/GSK3ß/ß-catenin signaling pathway in vivo and in vitro. Taken together, these findings unveiled the mechanism of mortalin in maintaining and regulating the stemness of BCSCs, and may offer novel therapeutic strategies for breast cancer treatment.

Medium-Term Clinical Results of High-Flexion Knee Prostheses in Patients with Rheumatoid Arthritis.

OBJECTIVE: This study was performed to evaluate the function and satisfaction outcome of patients with rheumatoid arthritis (RA) who underwent total knee arthroplasty (TKA) with high-flexion prostheses. MATERIALS AND METHODS: Twenty-two patients (35 knees) using high-flexion prostheses (Zimmer, Warsaw, IN) were followed up for a period of 7-11 years from February 2007 to December 2009. Clinical and radiographic follow-up was performed using Hospital for Special Surgery (HSS), Short-Form 36 scores (SF-36), American Knee Society score (KSS), and Knee Society Total Knee Arthroplasty Roentgenographic Evaluation and Scoring System. Patient satisfaction assessments took place at the final follow-up sessions using the Marsh Satisfaction Questionnaire. RESULTS: The average ROM improved from preoperative 68.43° ± 33.78° to 95.54° ± 7.03° at the final follow-up. The HSS score and KSS score for pain improved from (46.49 ± 12.73) points to (85.46 ± 3.90) points and from 20.57 ± 5.91 points to 47.43 ± 3.51 points at the follow-up evaluation, respectively. Physical Component Summary(PCS) and Physical Component Summary (MCS) scores were 45.38 and 52.56, respectively by the end of follow-up. Deep venous thrombosis developed in one patient and one patient required surgical revision due to infection. There were no instances of prosthetic loosening. The satisfaction rate of patients was 95.5%. CONCLUSION: Although this particular model of TKA did not yield high-flexion angles (ie, 140°) required for kneeling, squatting, or rising from the floor, significant clinical and radiographic gains were evident in these patients with RA.

Homoharringtonine inhibited breast cancer cells growth via miR-18a-3p/AKT/mTOR signaling pathway.

Homoharringtonine (HHT), a natural alkaloid derived from the cephalotaxus, exhibited its anti-cancer effects in hematological malignancies clinically. However, its pesticide effects and mechanisms in treating solid tumors remain unclear. In this study, we found that HHT was capable of inhibiting tumor growth after 5-days treatment of breast cancer cells, MCF-7, in vivo. Furthemore, HHT also significantly inhibited the cancer cell growth and induced cell apoptosis in vitro. miRNA sequencing proved miR-18a-3p was noticeably downregulated in the cells after HHT treatment. Moreover, downregulating miR-18a-3p increased HHT-induced cell apoptosis; our data supported that HHT suppressed miR-18a-3p expression and inhibited tumorigenesis might via AKT-mTOR signaling pathway. In conclusion: our study proved that HHT suppressed breast cancer cell growth and promoted apoptosis mediated by regulating of the miR-18a-3p-AKT-mTOR signaling pathway, HHT may be a promising antitumor agent in breast cancer treatment.

[Correlation between cold pain of knee joint and subchondral bone marrow edema in patients with knee osteoarthritis].

OBJECTIVE: To investigate relationship between cold pain of knee joint and subchondral bone marrow edema (BME). METHODS: From May 2018 to August 2019, 92 patients with knee osteoarthritis (KOA) associated with cold pain of knee were admitted, all patients were underwent MRI examination. The patients were divided into observation group (47 patients with BME) and control group(45 patients without BME). In observation group, there were 6 males and 41 females aged from 36 to 87 years old with an average of (63.2±12.3) years old. In control group, there were 10 males and 35 females, aged from 48 to 84 years old with an average of (62.7±8.3) years old. All patientswere treated with drugs. The degree of joint degeneration was evaluated by Kellgren-Lawrence (K-L) grading. Degree of cold pain of knee was evaluated by knee cold pain score, and degree of BME was evaluated according to WORMS. The correlation between cold pain of knee and K-L grading and BME was analyzed. RESULTS: Score of cold pain in observation group (15.55±7.68) was higher than that of control group (9.42± 5.50), which had significant difference (t=4.383, P<0.001). There was no correlation between cold pain of knee and K-L grading(χ2=2.138, P=0.907). There was correlation between BME grading and degree of cold pain in observation group(χ2=19.709, P<0.001), and Spearman correlation coefficient was rs=0.509(P<0.001). CONCLUSION: The cold pain of KOA patients is not related to K-L grading, but corelate with BME grading. The Cold pain of knee was more pronounced in KOA patients with BME, and the severity of BME is often related to degree of cold pain. It seemed to be a tendency:the more serious BME, the heavier coldpain.

Montelukast alleviates inflammation in experimental autoimmune encephalomyelitis by altering Th17 differentiation in a mouse model.

Montelukast is a leukotriene receptor antagonist that is known to prevent allergic rhinitis and asthma. Blocking the Cysteinyl leukotriene receptor (CysLTR1), one of the primary receptors of leukotrienes, has been demonstrated to be efficacious in ameliorating experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), through disrupting chemotaxis of infiltrating T cells. However, the role of CysLTR1 in the pathogenesis of MS is not well understood. Here, we show that MS patients had higher expression of CysLTR1 in the circulation and central nervous system (CNS). The majority of CD4+ T cells expressed CysLTR1 in MS lesions. Among T-cell subsets, Th17 cells had the highest expression of CysLTR1, and blocking CysLTR1 signalling abrogated their development in vitro. Inhibition of CysLTR1 by montelukast suppressed EAE development in both a prophylactic and therapeutic manner and inhibited myelin loss in EAE mice. Similarly, the in vivo results showed that montelukast inhibited Th17 response in EAE mice and that Th17 cells treated with montelukast had reduced encephalitogenic in adoptive EAE. Our findings strongly suggest that targeting Th17 response by inhibiting CysLTR1 signalling could be a promising therapeutic strategy for the treatment of MS and CNS inflammatory diseases.

Overcoming Intrinsic H3K27me3 Imprinting Barriers Improves Post-implantation Development after Somatic Cell Nuclear Transfer.

Successful cloning by somatic cell nuclear transfer (SCNT) requires overcoming significant epigenetic barriers. Genomic imprinting is not generally regarded as such a barrier, although H3K27me3-dependent imprinting is differentially distributed in E6.5 epiblast and extraembryonic tissues. Here we report significant enhancement of SCNT efficiency by deriving somatic donor cells carrying simultaneous monoallelic deletion of four H3K27me3-imprinted genes from haploid mouse embryonic stem cells. Quadruple monoallelic deletion of Sfmbt2, Jade1, Gab1, and Smoc1 normalized H3K27me3-imprinted expression patterns and increased fibroblast cloning efficiency to 14% compared with a 0% birth rate from wild-type fibroblasts while preventing the placental and body overgrowth defects frequently observed in cloned animals. Sfmbt2 deletion was the most effective of the four individual gene deletions in improving SCNT. These results show that lack of H3K27me3 imprinting in somatic cells is an epigenetic barrier that impedes post-implantation development of SCNT embryos and can be overcome by monoallelic imprinting gene deletions in donor cells.

Steaming Changes the Composition of Saponins of Panax notoginseng (Burk.) F.H. Chen That Function in Treatment of Hyperlipidemia and Obesity.

Saponins of Panax notoginseng (Burk.) F.H. Chen have been classified as a type of composition in functional foods for numerous diseases. However, its mild effects and other characteristics limited clinical applications in diseases. Inspired by "nine steaming and nine processing" of P. notoginseng in traditional Chinese medicine, we developed a "steaming"-mimic protocol, which significantly changed the composition of saponins of P. notoginseng from the original, R1, Rg1, Re, Rb1, and Rd (raw-PNS), to the products after steaming, 20S/R-Rh1, Rk3, Rh4, 20S/R-Rg3, Rk1, and Rg5 (N-PNS). Surprisingly, N-PNS demonstrated promising activities in improving hyperlipidemia and reducing body weight and weight of white adipose tissue and the inhibition of adipogenesis in obese mice. In accordance with the results in vivo, N-PNS remarkably blunted adipogenesis at the early stage of differentiation dose-dependently in vitro. Moreover, we demonstrated that the activity may involve the adenosine monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway by promoting phosphorylation of AMPKT172 and downregulating its downstream factors: sterol regulatory element binding protein 1c, stearoyl-CoA desaturase 1, and fatty acid synthase. Taken together, the steaming-induced eight compositions of saponins showed a very promising function in improving hyperlipidemia and obesity both in vivo and in vitro, providing fundamental evidence for future study and application in treatment of hyperlipidemia, obesity, and other lipid-related metabolic syndromes.

MicroRNA-552 promotes tumor cell proliferation and migration by directly targeting DACH1 via the Wnt/ß-catenin signaling pathway in colorectal cancer.

The purpose of the present study was to analyze the crucial role of microRNAs (miRNAs/miRs) involved in the proliferation and migration of colorectal cancer (CRC) and to investigate their underlying mechanisms. The present study discusses the expression and function of miR-552 in CRC. The expression level of miR-552 in CRC cells and tissues was observed, and it was suggested that the high expression of miR-552 accelerated the proliferation and migration of CRC cells in vitro. Notably, a result of the present study was that the cell fate determination factor Dachshund family transcription factor 1 (DACH1) was identified as a direct target of miR-552. Suppressing miR-552 expression in CRC cells increased endogenous DACH1 mRNA and protein levels, which was negatively correlated with miR-552. DACH1 performs an important role in the development of a number of neoplasms, and has the ability to regulate the Wnt/ß-catenin signaling pathway as a novel predictive and diagnostic biomarker. Accordingly, it was concluded that miR-552 exerted a tumor-promoting role in CRC development by targeting DACH1, which may contribute to the increase in the rates of CRC proliferation and migration. miR-552 may serve as a potential diagnostic and prognostic biomarker for CRC.

Thyroid surgery without antibiotic prophylaxis: experiences with 1,030 patients from a teaching hospital in China.

OBJECTIVE: Our objective was to evaluate the outcome of thyroidectomy without the use of prophylactic antibiotics. This study was held from January 2005 to May 2012 in a teaching hospital in Dongguan, China. METHODS: A total of 1,030 thyroidectomy patients were retrospectively reviewed and basic data were recorded, including age, sex, peri-operative antibiotic use, type of thyroid surgery done, and post-operative complications. Either an open approach or an endoscopic approach was performed according to the doctor's or patient's preference following a strict aseptic technique. The drain was routinely placed. Any complications were analyzed. RESULTS: A total of 834 (81 %) females and 196 (19 %) males were included, giving a ratio of 4.2:1. The average age was 38.3 years. The mean operation time was 85.3 min. Pathological type included 818 (79.4 %) nodular goiter, 34 (3.3 %) Graves' disease, 102 (9.9 %) nodular papillary hyperplasia, 12 (1.2 %) Hashimoto's disease, 62 (6 %) papillary carcinoma, and 2 (0.2 %) medullary carcinoma. Four patients had postoperative bleeding, four had temporally recurrent nerve paralysis. Only one had wound infection (0.09 %). CONCLUSION: Antibiotic prophylaxis in elective thyroidectomy is not an essential pre-operation preparation for all patients, if guidelines for antibiotic prophylaxis in clean surgery are adhered to and surgeons have sophisticated skills in the procedure.

[Isolation and characterization of human breast tumor stem cells].

AIM: To isolate and culture human breast cancer stem cells, and characterize their biological properties in vitro. METHODS: Fresh tumor tissues of breast cancer patients were collected from surgical rooms. The tissue samples were mechanically sheared and enzyme-digested to get single cell suspensions. Cancer cell surface antigens were analyzed by flow cytometry, and cell growth curve was established by MTT assay; Real-time quantitative PCR (qPCR) was employed to detect the expressions of stem cell-specific genes Sox2 and Nanog in the cancer cells; Immunofluorescence staining was performed to examine the expressions of breast cancer-specific proteins Bcl-2 and progesterone receptor (PR). RESULTS: Human breast cancer stem cells grew in the form of spheres. These stem cells had a CD44(+);/CD24(-); phenotype as characterized by flow cytometry, expressed high levels of Sox2 and Nanog genes, and were positive for Bcl-2 and PR. CONCLUSION: Human breast tumors contain CD44(+);/CD24(-); cancer stem cells which are capable of self-renewal and proliferation, and can be long-term cultured and expanded in vitro.

[Effects of IFN-γ on immunomodulation of mesenchymal stem cells isolated from fetal side of human placenta tissues].

AIM: To obtain fetal mesenchymal stem cells isolated from human placental tissue (hfPMSCs) and study the effects of IFN-γ on immunomodulatory property of hfPMSCs in vitro. METHODS: hfPMSCs were isolated from the fetal side of human placentas by enzyme digestion. Expressions of cell surface antigens CD73, CD90, CD105, CD14, CD34, CD45 and HLA-DR were tested by flow cytometry. The fetal origin of the placental cells was verified by DNA identity test using four known short tandem repeat (STR) polymorphic markers D2S1399, D10S2325, D18S535 and GATA198B05. After hfPMSCs were treated with 10 ng/mL IFN-γ, the expressions of IL-6, IL-8, IL-10 and HGF at both mRNA and protein levels were measured using quantitative real-time PCR (q-PCR) and ELISA. The expressions in controls without IFN-γ treatment were examined in the same way. RESULTS: The isolated cells expressed MSCs surface markers CD73, CD90 and CD105, but did not express CD14, CD34, CD45 and HLA-DR. For all the STR markers tested, the MSCs from the fetal side of the placenta carried the alleles of both maternal and paternal origin, indicative of the fetal genome. q-PCR indicated that the expression of IL6 mRNA in IFN-γ treated group was significantly higher than those in the control group (P<0.05), whereas the levels of IL-10 and HGF mRNA were down-regulated by IFN-γ treatment (P<0.05). ELISA revealed that IFN-γ treated group expressed strikingly the higher level of IL-6 protein compared with the control group (P<0.05), and the lower levels of IL-8, IL-10 and HGF (P<0.05). CONCLUSION: The method used in the study can obtain hfPMSCs from human placenta tissues, and IFN-γ has a negative effect on immune suppression function of the hfPMSCs.