Synchrotron Infrared Microspectroscopy for Stem Cell Research.

Stem cells have shown great potential functions for tissue regeneration and repair because of their unlimited self-renewal and differentiation. Stem cells reside in their niches, making them a hotspot for the development and diagnosis of diseases. Complex interactions between niches and stem cells create the balance between differentiation, self-renewal, maturation, and proliferation. However, the multi-facet applications of stem cells have been challenged since the complicated responses of stem cells to biological processes were explored along with the limitations of current systems or methods. Emerging evidence highlights that synchrotron infrared microspectroscopy, known as synchrotron radiation-based Fourier transform infrared microspectroscopy, has been investigated as a potentially attractive technology with its non-invasive and non-biological probes in stem cell research. With their unique vibration bands, the quantitative mapping of the content and distribution of biomolecules can be detected and characterized in cells or tissues. In this review, we focus on the potential applications of synchrotron infrared microspectroscopy for investigating the differentiation and fate determination of stem cells.

Mid- and Long-Term Outcome Comparisons of Everolimus-Eluting Bioresorbable Scaffolds Versus Everolimus-Eluting Metallic Stents: A Systematic Review and Meta-analysis.

BACKGROUND: Percutaneous coronary interventions to implant bioresorbable vascular scaffolds (BVSs) were designed to reduce the late thrombotic events that occur with metallic stents. PURPOSE: To estimate the incidence of scaffold thrombosis after BVS implantation and compare everolimus-eluting BVSs with everolimus-eluting metallic stents (EESs) in terms of safety and efficacy at mid- and long-term follow-up in adults who had a percutaneous coronary intervention. DATA SOURCES: PubMed, EMBASE, the Cochrane Library, conference proceedings, and relevant Web sites from inception until 20 May 2017, without language restriction. STUDY SELECTION: 7 randomized trials and 38 observational studies (each with a minimum of 6 months and 100 patient-years of follow-up) in adults with coronary artery disease who had a BVS or an EES and reported scaffold or stent thrombosis (main outcome) or other secondary outcomes (such as death, myocardial infarction, or revascularization). DATA EXTRACTION: 2 reviewers independently extracted study data, rated study quality, and assessed strength of evidence. DATA SYNTHESIS: The pooled incidence of definite or probable scaffold thrombosis after BVS implantation was 1.8% (95% CI, 1.5% to 2.2%) at a median follow-up of 1 year (41 studies, 21 884 patients) and 0.8% (CI, 0.5% to 1.3%) beyond 1 year (14 studies, 4688 patients). Seven trials involving 5578 patients that directly compared BVSs with EESs showed an increased risk for definite or probable scaffold thrombosis (odds ratio [OR], 3.40 [CI, 2.01 to 5.76]) with BVSs at a median follow-up of 25 months. Increased risks were present at early (prominently subacute), late, and very late stages, and odds beyond 1 year were almost double those seen within 1 year. Bioresorbably vascular scaffolds increased risks for myocardial infarction (OR, 1.63 [CI, 1.26 to 2.10]), target lesion revascularization (OR, 1.31 [CI, 1.03 to 1.67]), and target lesion failure (OR, 1.37 [CI, 1.12 to 1.66]); the odds for these 3 end points also increased over time. The incidences of all-cause, cardiac, and noncardiac death and of target vessel and any revascularization did not differ. LIMITATION: Quality of observational studies was unclear, and some data were unpublished. CONCLUSION: Compared with EESs, BVSs increased the risks for scaffold thrombosis and other thrombotic events at mid- and long-term follow-up, and risks increased over time. PRIMARY FUNDING SOURCE: National Natural Science Foundation of China.

Optimization of paeonol-loaded microparticle formulation by response surface methodology.

In this study, a central composite rotatable design based on response surface methodology (RSM) was employed to design and formulate an appropriate paeonol microparticle formulation. Five levels of a three-factor, rotatable, central composite design were used to evaluate the critical formulation variables. The optimum conditions for preparing paeonol-loaded microparticles were predicted to be: polyvinyl alcohol (PVA) content (2.84%), the ratio of drug to polymer (6.88) and the stirring rate (1007.59 rpm). The optimized responses for production yield and loading efficiency were found to be 68.86% and 55.90%, respectively, and the particle size were 23.27 ± 0.76 µm and the sorting coefficient (σ) was 0.732. Furthermore, in vitro release study suggested that microparticle could be a suitable delivery system in treating skin disease for its sustained release of drug. In conclusion, RSM can be successfully used to optimize the effect of formulation variables.

[Preparation of paeonol transdermal delivery systems based on proniosomes-based ointment and its pharmacokinetics characters].

The paeonol proniosomes ointment and ordinary ointment were administered to rats. Physiological saline served as perfused solution. The perfusion rate was 5 mL x L(-1) and the microdialysis samples were collected every 20 min intervals. The paeonol concentration in perfused solution was determined by HPLC. Investigation of the pharmacokinetics of paeonol proniosomes ointment and ordinary ointment by the skin-blood synchronous microdialysis coupled with HPLC is reported in this study. The results show that the recovery was (54.80 +/- 1.50)% in vitro and (54.58 +/- 4.61)% in vivo. The results showed that paeonol proniosomes ointment significantly raised the drug concentrations in skin more than the paeonol ordinary ointment. The paeono proniosomes ointment has less drugs into the blood as the ordinary ointments in blood, but its blood drug concentrations were steadier. The paeonol proniosomes ointment may be developed into a new preparation.

Transcription factor Dmrt1 triggers the SPRY1-NF-κB pathway to maintain testicular immune homeostasis and male fertility.

Bacterial or viral infections, such as Brucella, mumps virus, herpes simplex virus, and Zika virus, destroy immune homeostasis of the testes, leading to spermatogenesis disorder and infertility. Of note, recent research shows that SARS-CoV-2 can infect male gonads and destroy Sertoli and Leydig cells, leading to male reproductive dysfunction. Due to the many side effects associated with antibiotic therapy, finding alternative treatments for inflammatory injury remains critical. Here, we found that Dmrt1 plays an important role in regulating testicular immune homeostasis. Knockdown of Dmrt1 in male mice inhibited spermatogenesis with a broad inflammatory response in seminiferous tubules and led to the loss of spermatogenic epithelial cells. Chromatin immunoprecipitation sequencing (ChIP-seq) and RNA sequencing (RNA-seq) revealed that Dmrt1 positively regulated the expression of Spry1, an inhibitory protein of the receptor tyrosine kinase (RTK) signaling pathway. Furthermore, immunoprecipitation-mass spectrometry (IP-MS) and co-immunoprecipitation (Co-IP) analysis indicated that SPRY1 binds to nuclear factor kappa B1 (NF-κB1) to prevent nuclear translocation of p65, inhibit activation of NF-κB signaling, prevent excessive inflammatory reaction in the testis, and protect the integrity of the blood-testis barrier. In view of this newly identified Dmrt1- Spry1-NF-κB axis mechanism in the regulation of testicular immune homeostasis, our study opens new avenues for the prevention and treatment of male reproductive diseases in humans and livestock.

Finding minimum gene subsets with heuristic breadth-first search algorithm for robust tumor classification.

BACKGROUND: Previous studies on tumor classification based on gene expression profiles suggest that gene selection plays a key role in improving the classification performance. Moreover, finding important tumor-related genes with the highest accuracy is a very important task because these genes might serve as tumor biomarkers, which is of great benefit to not only tumor molecular diagnosis but also drug development. RESULTS: This paper proposes a novel gene selection method with rich biomedical meaning based on Heuristic Breadth-first Search Algorithm (HBSA) to find as many optimal gene subsets as possible. Due to the curse of dimensionality, this type of method could suffer from over-fitting and selection bias problems. To address these potential problems, a HBSA-based ensemble classifier is constructed using majority voting strategy from individual classifiers constructed by the selected gene subsets, and a novel HBSA-based gene ranking method is designed to find important tumor-related genes by measuring the significance of genes using their occurrence frequencies in the selected gene subsets. The experimental results on nine tumor datasets including three pairs of cross-platform datasets indicate that the proposed method can not only obtain better generalization performance but also find many important tumor-related genes. CONCLUSIONS: It is found that the frequencies of the selected genes follow a power-law distribution, indicating that only a few top-ranked genes can be used as potential diagnosis biomarkers. Moreover, the top-ranked genes leading to very high prediction accuracy are closely related to specific tumor subtype and even hub genes. Compared with other related methods, the proposed method can achieve higher prediction accuracy with fewer genes. Moreover, they are further justified by analyzing the top-ranked genes in the context of individual gene function, biological pathway, and protein-protein interaction network.

A New Distance Stereotest by Autostereoscopic Display Using an Eye-Tracking Method.

Objectives: This research aimed to present a novel glasses-free distance random-dot stereotest system (GFDRDSS) using an eye-tracking method. Methods: A single-view autostereoscopic display applying a backlight control system combined with an eye-tracking method and the corresponding random-dot stereotest software were developed to create a GFDRDSS with a viewing distance of 5 m. The stereoacuity of 12 subjects with normal eye position was evaluated using the Randot Stereotest, Stereoscopic Test Charts vol. 3 (Yan's Charts), Distance Randot® Stereotest, and GFDRDSS. Results: The GFDRDSS could provide distinct and stable glasses-free stereoscopic perception even while the subject was moving their head. It could evaluate binocular disparities of 40-2,400 arcsec. Eleven subjects with normal near visual acuity had fine near stereovision (20-60 arcsec) using the Randot stereotest and Yan's Charts. Under refractive correction, 10 subjects had fine stereovision (≤60 arcsec) using the GFDRDSS at a distance of 5 m, and 9 had fine stereovision using the Distance Randot® Stereotest at 3 m. Other subjects described the 100 arcsec-level stereograms correctly. The results exhibited a concordance of stereoacuity within one degrade between the two distance stereotests. Conclusion: The proposed GFDRDSS can alternately project a couple of random-dot stereograms to the subjects' eyes and provide a glasses-free distance stereotest, which showed good concordance with the Distance Randot® Stereotest. More data are needed for statistical studies.

Characteristics of temporomandibular joint vibrations in anterior disk displacement with reduction in adults.

The objective to the present study was to compare temporomandibular joint (TMJ) vibration in anterior disk displacement with reduction (ADDWR) in adults and to explore the diagnostic value of frequency spectrography of TMJ vibrations. Twenty-one patients with ADDWR formed the case group and were further divided into three subgroups according to the degree of disk displacement, and 26 symptomless adults formed the control group. The joint vibration was recorded during rhythmic maximal open-close jaw movement using JVA/JT, BioPAK (Bioresearch, Inc., Brown Deer, WI)). The sensitivity and specificity of the total integral for diagnosis of ADDWR was calculated. All TMJ vibration parameters, including total integral, integral>300Hz, integral<300Hz, >300/<300 ratio, peak amplitude, peak and median frequencies of the case group were significantly higher than that of the control group. Along with the degree of disk displacement, the amplitude and frequency of TMJ vibrations increased, and the total integral significantly increased. The total integral demonstrated high sensitivity and specificity for diagnosis of ADDWR (85.7% and 84.6%, respectively). TMJ vibration was significantly higher in adults with ADDWR than that in the symptomless control group. Different pathological stages of disk displacement have different TMJ vibrations.

[Role of perforin in severe preeclampsia].

OBJECTIVES: To investigate the possible role of perforin (PFN) in the pathogenesis of severe preeclampsia. METHODS: Thirty-two cases of severe preeclampsia were included in the study. Thirty-two cases of normal pregnancy were selected as control group in random. The expression of PFN mRNA in the peripheral blood mononuclear cells (PBMC) was detected by reverse transcription (RT)-PCR, and its correlation with mean arterial pressure was analyzed in severe preeclamptic patients. The expression of PFN protein in the decidua was detected by immunohistochemistry. RESULTS: (1) The expression of PFN mRNA in PBMC:the PFN mRNA level in severe preeclamptic group was 1.19 ± 0.31, and that in normal pregnancy group is 0.82 ± 0.28. The PFN mRNA level in severe preeclamptic group was significantly higher than that of control group (P < 0.01). (2) Correlation analysis: the mean blood pressure in severe preeclampsia group was (133 ± 5) mm Hg (1 mm Hg = 0.133 kPa). There was significant positive correlation between level of PFN mRNA in PBMC and mean blood pressure in severe preeclamptic patients (r = 0.701, P = 0.000). (3) Decidual PFN protein expression:PFN protein was mainly expressed in lymphocytes and the cytoplasm of decidual stromal cells. The positive ratio of PFN in the decidua of severe preeclamptic patients was 84% (27/32), significantly higher than that of control group (53%, 17/32, P < 0.01). CONCLUSIONS: Expression of PFN was significantly increased in severe preeclampsia, and it was of significant positive correlation with mean blood pressure. PFN may participate in the pathogenesis of severe preeclampsia.

Treatment of ammonia-embodied wastewater by a transition-metal-based photochemical catalysis strategy.

Inspired by the self-purification process and a low nitrogen content of the ocean, and the fact that the driving-force behind ecological cycle is solar irradiation, a novel photochemical strategy was designed to spontaneously remove inorganic ammonia nitrogen from wastewater with solar irradiation. This strategy is based on the principles of green chemistry and energy efficiency, and meanwhile the prevention from the introduction of accompanying pollution. In our strategy, a photo-Fe (or Mn)-O2 system was built to remove ammonia-nitrogen from its aqueous solution. The results show that with full band solar irradiation at a range of 10-30 mW cm-2, in weak alkaline condition, more than 90% of ammonia-nitrogen can be effectively removed from NH4Cl aqueous solution by the new strategy, with a residual concentration as low as 2 mg L-1. Mn(III) was proved to be a better catalyst than Fe(III). The catalytic mechanism of N-removal is the generation of •OH during the process of the photoreduction of transition metal hydroxides. DFT theory had been applied to help explaining the mechanism. Different from general knowledge, in our strategy, an alkaline environment, where the generation rate of radicals was relatively slow and comparable to oxidation rate of transition metal ions, can guarantee the stability and persistency of the catalytic reaction. No NOx was produced in this strategy. This new strategy provides a new possibility of cost-efficient and environmental-friendly wastewater treatment, and has certain meaning of understanding how self-purification works in nature.

Body Mass Index: An Effective Predictor of Ejection Fraction Improvement in Heart Failure.

Background: Heart failure patients with higher body mass index (BMI) exhibit better clinical outcomes. Therefore, we assessed whether the BMI can predict left ventricular ejection fraction (EF) improvement following heart failure. Methods and Results: We included 184 patients newly diagnosed with dilated cardiomyopathy and reduced EF in our center and who underwent follow-up examination of EF via echocardiography after 6 months. The EF improved at 6 months in 88 participants, who were included in the heart failure with recovered EF (HFrecEF) subgroup. Patients in whom the EF remained reduced were included in the heart failure with persistently reduced EF (persistent HFrEF) subgroup. Our analyses revealed that EF increase correlated with age (r = -0.254, P = 0.001), left ventricular diastolic dimension (LVDD; r = -0.210, P = 0.004), diabetes (P = 0.034), brain natriuretic peptide (r = -0.199, P = 0.007), and BMI grade (P = 0.000). BMI grade was significantly associated with elevated EF after adjustment for other variables (P = 0.001). On multivariable analysis, compared to patients with persistent HFrEF, those with HFrecEF had higher BMI [odds ratio (OR) = 2.342 per one standard deviation increase; P = 0.001] and lower LVDD (OR = 0.466 per one standard deviation increase; P = 0.001). ROC-curve analysis data showed that BMI > 22.66 kg/m2 (sensitivity 84.1%, specificity 59.4%, AUC 0.745, P = 0.000) indicate high probability of EF recovery in 6 months. Conclusions: Our data suggest that higher BMI is strongly correlated with the recovered EF and that BMI is an effective predictor of EF improvement in patients with heart failure and reduced EF.

Single-cell RNA sequencing reveals atlas of dairy goat testis cells.

Single-cell RNA sequencing (scRNA-seq) is useful for exploring cell heterogeneity. For large animals, however, little is known regarding spermatogonial stem cell (SSC) self-renewal regulation, especially in dairy goats. In this study, we described a high-resolution scRNA-seq atlas derived from a dairy goat. We identified six somatic cell and five spermatogenic cell subtypes. During spermatogenesis, genes with significantly changed expression were mainly enriched in the Notch, TGF-ß, and Hippo signaling pathways as well as the signaling pathway involved in the regulation of stem cell pluripotency. We detected and screened specific candidate marker genes ( TKTL1 and AES) for spermatogonia. Our study provides new insights into goat spermatogenesis and the development of testicular somatic cells.

Predicting protein-protein interactions from sequence using correlation coefficient and high-quality interaction dataset.

Identifying protein-protein interactions (PPIs) is critical for understanding the cellular function of the proteins and the machinery of a proteome. Data of PPIs derived from high-throughput technologies are often incomplete and noisy. Therefore, it is important to develop computational methods and high-quality interaction dataset for predicting PPIs. A sequence-based method is proposed by combining correlation coefficient (CC) transformation and support vector machine (SVM). CC transformation not only adequately considers the neighboring effect of protein sequence but describes the level of CC between two protein sequences. A gold standard positives (interacting) dataset MIPS Core and a gold standard negatives (non-interacting) dataset GO-NEG of yeast Saccharomyces cerevisiae were mined to objectively evaluate the above method and attenuate the bias. The SVM model combined with CC transformation yielded the best performance with a high accuracy of 87.94% using gold standard positives and gold standard negatives datasets. The source code of MATLAB and the datasets are available on request under smgsmg@mail.ustc.edu.cn.

Neighborhood rough set reduction-based gene selection and prioritization for gene expression profile analysis and molecular cancer classification.

Selection of reliable cancer biomarkers is crucial for gene expression profile-based precise diagnosis of cancer type and successful treatment. However, current studies are confronted with overfitting and dimensionality curse in tumor classification and false positives in the identification of cancer biomarkers. Here, we developed a novel gene-ranking method based on neighborhood rough set reduction for molecular cancer classification based on gene expression profile. Comparison with other methods such as PAM, ClaNC, Kruskal-Wallis rank sum test, and Relief-F, our method shows that only few top-ranked genes could achieve higher tumor classification accuracy. Moreover, although the selected genes are not typical of known oncogenes, they are found to play a crucial role in the occurrence of tumor through searching the scientific literature and analyzing protein interaction partners, which may be used as candidate cancer biomarkers.

A protein interaction network analysis for yeast integral membrane protein.

Although the yeast Saccharomyces cerevisiae is the best exemplified single-celled eukaryote, the vast number of protein-protein interactions of integral membrane proteins of Saccharomyces cerevisiae have not been characterized by experiments. Here, based on the kernel method of Greedy Kernel Principal Component analysis plus Linear Discriminant Analysis, we identify 300 protein-protein interactions involving 189 membrane proteins and get the outcome of a highly connected protein-protein interactions network. Furthermore, we study the global topological features of integral membrane proteins network of Saccharomyces cerevisiae. These results give the comprehensive description of protein-protein interactions of integral membrane proteins and reveal global topological and robustness of the interactome network at a system level. This work represents an important step towards a comprehensive understanding of yeast protein interactions.

Cardiovascular Safety, Long-Term Noncardiovascular Safety, and Efficacy of Sodium-Glucose Cotransporter 2 Inhibitors in Patients With Type 2 Diabetes Mellitus: A Systemic Review and Meta-Analysis With Trial Sequential Analysis.

BACKGROUND: The cardiovascular and long-term noncardiovascular safety and efficacy of SGLT2 (sodium-glucose cotransporter 2) inhibitors have not been well documented. METHODS AND RESULTS: For cardiovascular outcomes, we performed a meta-analysis with trial sequential analysis of randomized controlled trials and adjusted observational studies, each with a minimum of 26 weeks and 2000 patient-years of follow-up. For long-term noncardiovascular safety and efficacy outcome analyses, we included only randomized controlled trials with at least 2 years and 1000 patient-years of follow-up. Five studies with 351 476 patients were included in cardiovascular outcomes analysis. Meta-analyses showed that SGLT2 inhibitors significantly reduced the risks of major adverse cardiac events (hazard ratio [HR]: 0.80; 95% confidence interval [CI], 0.69-0.92; P=0.002), all-cause mortality (HR: 0.67; 95% CI, 0.54-0.84; P<0.001), cardiovascular mortality (HR: 0.77; 95% CI, 0.60-0.98; P=0.03), nonfatal myocardial infarction (HR: 0.86; 95% CI, 0.76-0.98; P=0.02), hospitalization for heart failure (HR: 0.62; 95% CI, 0.55-0.69; P<0.001), and progression of albuminuria (HR: 0.68; 95% CI, 0.58-0.81; P<0.001). No significant difference in nonfatal stroke was found. Analyses limited to randomized controlled trials showed similar findings. Trial sequential analysis provided firm evidence of a 20% reduction in major adverse cardiac events, all-cause mortality, and hospitalization for heart failure with SGLT2 inhibitors, but evidence remains inconclusive for cardiovascular mortality. Nine randomized controlled trials contributed to long-term noncardiovascular and efficacy analyses. SGLT2 inhibitors reduced incidence of hypoglycemia and acute kidney injury but increased the risks of urinary tract and genital infections. CONCLUSIONS: SGLT2 inhibitors showed remarkable cardiovascular- and renal-protective effects and good long-term noncardiovascular safety with sustained efficacy.

Expression of the SRY-related HMG box protein SOX2 in human gastric carcinoma.

SOX2, a SRY-related HMG box protein, is thought to be an important transcription factor during organogenesis, including the stomach although the expression and function are unclear. We investigated SOX2 protein expression to clarify its roles in differentiation and carcinogenesis of the stomach. Using polyclonal SOX2 antibodies, expression of SOX2 in gastric normal mucosae, intestinal metaplasia and carcinomas from 68 gastric carcinoma patients was studied by immuohistochemistry. SOX2 was strongly and moderately expressed in the nuclei of the foveolar epithelium and intestinal metaplasia, respectively, the expression being much higher than that in carcinomas (p<0.0001). Using antibodies to MUC5AC, MUC2 and CD10, the 68 gastric carcinomas were classified into gastric type, intestinal type, mixed gastric and intestinal type, and null type. A significant difference in SOX2 expression was observed between the gastric and intestinal types (p<0.05), with a higher expression in the former than in the latter. Moreover, over-expression of SOX2 induced the mRNA expression of endogenous MUC5AC, a specific mucin marker for the gastric type, in COS-7 cells. Our findings indicate that SOX2 may play a role in differentiation of the human gastric epithelium, and that SOX2 may be involved in gastric carcinogenesis, particularly in the gastric type.

[HPRT gene knock-out from rat fetal neural stem cells].

The 3.0 kb 5' arm (long arm, LA) of rat HPRT gene knock-out vector was cut by Sal I from rat HPRT gene genomic bacterial artificial chromosome (BAC) , and the 1.7 kb 3' arm (short arm, SA) was proliferated by PCR. Neo', 5' arm, 3' arm were sequentially cloned into pBS vector's relative restriction enzyme sites. For acquirement of tk gene, the 5' arm -Neo' -3' arm fragment inserted into pKO vector to construct pKO-HPRT. The pKO-HPRT was linearized by Not I, extracted by ethidium bromide, butanol and phenol/chloroform, and dialyzed by 0.025 microlmol/L Millipore. At the same time, rat neural stem cells cultured from E14.5-16.5 rat fetal brain. Passage 2 rFNSCs was tranfected by linearized pKO-HPRT with Fugene-6t transfection reagents. After 80 microg/ml G418 and 0.2 micromol/L ganciclovir selection, the survived cells was cultured in suspension to form neural spheres. The spheres can be picked up under the microscopy, and proliferated in 96- ,48- and 24-well plates sequentially. When the cell number reached 4 x 10(3)/well, half cells was lysed by lysis buffer to extract DNA, the other half was kept on growing to freeze and extract RNA. The knock-out cell colonies first detected by PCR, then confirmed by Southern blot and RT-PCR. All the results show that we have knocked out HPRT gene in three rat fetal neural stem cell colonies from 32 colonies.

[Studies of effects of lipid on rat fetal neural stem cells].

Rat fetal neural stem cells (rFNSCs) was separated from embryo about 14.5-16.5 days, and cultured in DMEM/F12 media with additives and epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Effects of lipid on growth and proliferation of rFNSCs was examined by counting the number of neurospheres and incorporation of 3H. The data show that chemical defined lipid improved rFNSCs' growth and cell division. Lipid will be another neural stem cell's culture media's additive.