Kaempferol mitigates sepsis-induced acute lung injury by modulating the SphK1/S1P/S1PR1/MLC2 signaling pathway to restore the integrity of the pulmonary endothelial cell barrier.

Sepsis-induced acute lung injury (SALI) is the common complication of sepsis, resulting in high incidence and mortality rates. The primary pathogenesis of SALI is the interplay between acute inflammation and endothelial barrier damage. Studies have shown that kaempferol (KPF) has anti-sepsis properties. Sphingosine kinase 1 (SphK1)/sphingosine-1-phosphate (S1P) signaling pathway's significance in acute lung damage and S1P receptor 1 (S1PR1) agonists potential in myosin light chain 2 (MLC2) phosphorylation are documented. Whether KPF can regulate the SphK1/S1P/S1PR1/MLC2 signaling pathway to protect the lung endothelial barrier remains unclear. This study investigates the KPF's therapeutic effects and molecular mechanisms in repairing endothelial cell barrier damage in both LPS-induced sepsis mice and human umbilical vein endothelial cells (HUVECs). KPF significantly reduced lung tissue damage and showed anti-inflammatory effects by decreasing IL-6 and TNF-α synthesis in the sepsis mice model. Further, KPF administration can reduce the high permeability of the LPS-induced endothelial cell barrier and alleviate lung endothelial cell barrier injury. Mechanistic studies showed that KPF pretreatment can suppress MLC2 hyperphosphorylation and decrease SphK1, S1P, and S1PR1 levels. The SphK1/S1P/S1PR1/MLC2 signaling pathway controls the downstream proteins linked to endothelial barrier damage, and the Western blot (WB) showed that KPF raised the protein levels. These proteins include zonula occludens (ZO)-1, vascular endothelial (VE)-cadherin and Occludin. The present work revealed that in mice exhibiting sepsis triggered by LPS, KPF strengthened the endothelial barrier and reduced the inflammatory response. The SphK1/S1P/S1PR1/MLC2 pathway's modulation is the mechanism underlying this impact.

Pink noise: a potential sound therapy for tinnitus.

At present, traditional music therapy based on "white noise" is commonly used to treat tinnitus patients. This study aims to apply "pink noise", which has a more balanced frequency distribution than white noise, to the field of tinnitus rehabilitation treatment to study its inhibitory effect on tinnitus. Fractal technology (fractal mathematics) was used to generate semi-repetitive tones as the basic tone of music therapy. This clinical study recruited 43 adult tinnitus patients with varying degrees of hearing loss as the study subjects. All patients received fractal music therapy with hearing aids. The study evaluated the Tinnitus Handicap Inventory (THI) of patients' usage and tested the indicators with paired sample T-tests. The results indicate that "pink noise" is effective in the treatment of tinnitus, and the differences in the scores are statistically significant before and after treatment (P<0.05). In addition, by comparing the tone spectra of "white noise" and "pink noise", we found that the frequency component power of "pink noise" is mainly concentrated in the middle and low frequency range. Its slope is -3 dB/Oct, which means that the frequency decays downwards as the ratio increases. In this study, a tinnitus music therapy based on the "pink noise" tone was developed, which mainly solved the awkward problem of no specific drugs or special tinnitus treatment equipment. At the same time, it helps to accelerate the development of related medical devices and improve the quality of life of tinnitus patients.

Evaluation of synovial inflammation in juvenile idiopathic arthritis using superb microvascular imaging compared with power Doppler ultrasound.

OBJECTIVES: This study aimed to investigate the diagnostic and prognostic performance of superb microvascular imaging (SMI) in evaluation of synovial inflammation in patients with juvenile idiopathic arthritis (JIA) compared with power Doppler ultrasound (PDUS). METHODS: Fifty-nine patients with active disease and 62 patients with inactive disease were enrolled. The synovial inflammation was evaluated via vascularity index (VI) of SMI and PDUS. The correlations between VIs and the inflammatory biomarkers were analysed by Spearman's coefficient. Receiver operating characteristics curves were plotted to examine the prognostic value of SMI and PDUS. RESULTS: The VI of SMI was significantly higher than that of PDUS in JIA patients regardless of the disease activity. The SMI and PDUS VI were significantly correlated with levels of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and serum amyloid A (SAA). The SMI VI was significantly higher in patients with relapse than in those with remission, and showed superior performance in predicting relapse in JIA patients with inactive disease. CONCLUSIONS: SMI may detect the synovial inflammation with greater sensitivity than PDUS in patients with JIA, and correlate well with the inflammatory biomarkers. SMI signal in the knees might play an important role in prediction of relapse in clinically inactive patients, thus allowing personalised treatment strategies for JIA patients.

Immune checkpoint inhibitor therapy-induced autoimmune polyendocrine syndrome type II and Crohn's disease: A case report.

BACKGROUND: The development of immune checkpoint inhibitors (ICIs) has heralded a new era in cancer treatment, enabling the possibility of long-term survival in patients with metastatic disease. Unfortunately, ICIs are increasingly implicated in the development of autoimmune diseases. CASE SUMMARY: We present a man with squamous cell carcinoma of the oropharynx on a combination of teriprizumab, docetaxel, and cisplatin therapy who developed autoimmune polyendocrine syndrome type II (APS-2) including thyroiditis and type 1 diabetes mellitus and Crohn's disease (CD). He developed thirst, abdominal pain, and fatigue after two-week treatment with the protein 1 ligand inhibitor teriprizumab. Biochemistry confirmed APS-2 and thyrotoxicosis. He was commenced on an insulin infusion. However, his abdominal pain persisted. Follow-up surgery confirmed CD and his abdominal pain was relieved by mesalazine. He was continued on insulin and mesalazine therapy. CONCLUSION: Immunotherapy can affect all kinds of organs. When clinical symptoms cannot be explained by a single disease, clinicians should consider the possibility of multisystem damage.

Effects of extreme pH conditions on the stability of As(V)-bearing schwertmannite.

Schwertmannite (Sch) is known to be an effective scavenger of arsenic (As) due to its strong binding affinity for toxic As species. However, the evolution of As-bearing schwertmannite under extreme pH conditions is poorly understood. In this study, we investigated the effects of extremely acidic and alkaline conditions on the stability of schwertmannite with structurally incorporated As(V) (CO-Sch) and schwertmannite with adsorbed As(V) (AD-Sch). The results show that both extremely acidic and alkaline conditions have significant effects on the evolution of minerals and liberation of iron and sulfate. At extremely acidic pH, the maximal release of ferric iron (Fe(III)) and sulfate from CO-Sch were greater than that from AD-Sch, whereas 6.2% and 0.3% of total As released from AD-Sch and CO-Sch, respectively. At extremely alkaline pH, aqueous Fe(III) was not observed, and Fe(III) was retained in As-bearing schwertmannite due to the chemical equilibrium between the dissolution of schwertmannite and re-precipitation of goethite; structurally incorporated As(V) promoted the liberation of sulfate. In addition, the adsorbed As on schwertmannite is more stable, which led to a minor release of As (0.8%) over a 30-d period, however, the liberated As(V) from CO-Sch accounts for up to 3.2%. Under extremely acidic and alkaline conditions, portions of AD-Sch and CO-Sch transformed from schwertmannite to goethite after 30 d, while schwertmannite was still the dominant mineral. Adsorbed As(V) inhibited the transformation of As-bearing schwertmannite to goethite more significantly than structurally incorporated As(V).

Antibody-sandwich ELISA analysis of a novel blood biomarker of CST4 in gastrointestinal cancers.

BACKGROUND: Members of the cystatin family have increasingly been proven to be involved in several tumors, including gastric cancer (GC) and colorectal cancer (CRC). Cystatin S (CST4) was found to be upregulated at the gene expression level in GC cells, making it a potential novel biomarker for the early diagnosis of gastrointestinal cancer. MATERIALS AND METHODS: Quantitative real-time polymerase chain reaction and Western blotting analysis were used to explore CST4 expression in gastrointestinal cancer tissues and cell lines. We purified CST4 recombinant protein and generated anti-CST4 monoclonal antibodies to develop an antibody-sandwich enzyme-linked immunosorbent assay (ELISA) analysis system for blood CST4 detection. The performance and clinical efficacy of the detection method were evaluated using a training set and validation set, respectively. RESULTS: According to the quantitative real-time polymerase chain reaction and Western blotting results, CST4-mRNA expression and protein expression were upregulated in gastrointestinal cancer tissues and cell lines. The ELISA detection system for CST4 showed significantly better sensitivities of 69.0% and 69.0% and specificities of 85.6% and 83.6% for GC and CRC, respectively, than other common clinical biomarkers, carcinoembryonic antigen, CA19-9, CA125, and CA72-4. Clinical verification experiments using GC and CRC validation sets also found distinguishable CST4 median concentrations (177.7 pg·mL-1 and 174.2 pg·mL-1 respectively) and high positive detection rates (72.3% and 88.4% respectively), further confirming the specificity and sensitivity of this method. CONCLUSION: We validated the overexpression of CST4 in gastrointestinal cancer tissues and cell lines and developed an antibody-sandwich ELISA analysis system for blood CST4 detection, which exhibited high specificity and sensitivity. Novel blood biomarkers of CST4 have enormous potential in terms of clinical diagnostic value in GC and CRC.

Development of hypoxia control device for cell culture in vitro / 口腔疾病防治

Objective@#To simulate the hypoxic environment of cells in vivo more approximately, a hypoxia control device which can control oxygen concentration automatically for cell culture in vitro was developed. @* Methods@#In vitro cell cultur e hypoxia control device consisted of hardware and software which were controlled by computer. The hardware was composed of oxygen detection and control system, nitrogen gas pipeline, cell culture and anti-jamming device, and the software was based on Clanguage. When oxygen controlle rreads the oxygen concent ration more than the set value, the nitrogen gas pipeline opens and nitrogen enter into the incubator; when reaching the set value, the nitrogen gas pipeline closes and nitrogen is not allowed to pass through. Circularly, a constant oxygen concentration was reached and maintained. This device was further verified by comparing the set value of the equipment with figures of the oxygen meter and by monitoring fluctuation condition after the oxygen concentration reaching the set value. @* Results @# There was no significant difference between the oxygen concentration set in the in vitro cell culture hypoxia control device and the actual oxygen concentration in the measured box (P > 0.05), and the accuracy of the device to set the value of the simulation up to ± 0.5% of the requirements to meet the system accuracy requirements, and can remain stable for a long time. @*Conclusion @#The device can effectively control the oxygen concentration to the required requirements, to meet the conditions in the hypoxic conditions for in vitro cell culture.

Infarcted cardiac microenvironment may hinder cardiac lineage differentiation of human embryonic stem cells.

Microenvironment regulates cell fate and function. In this study, we investigated the effects of the infarcted cardiac microenvironment on cardiac differentiation of human embryonic stem cells (hESCs). hESCs were intramyocardially transplanted into infarcted or uninjured rat hearts. After 4 weeks, mesodermal and cardiac lineage markers were detected by immunofluorescence. Cardiac function was assessed by echocardiography. hESCs were differentiated in vitro under hypoxic (5% O2 ), low-nutrient (5% FBS), or control condition. The numbers of beating clusters, proportions of cardiac troponin T (cTnT)-positive cells, and relative levels of cardiac-specific markers were determined. Results showed that in both uninjured and infarcted hearts, hESCs survived, underwent development, and formed intracardiac grafts, with a higher proportion in the uninjured hearts. However, cells that were double positive for human fetal liver kinase 1 (Flk1), a marker of cardiac progenitors, and human ß-tubulin, a marker for labeling human cells, were found in the uninjured hearts but not in the infarcted hearts. hESC transplantation did not restore the cardiac function of acutely infarcted rats. In vitro, low FBS treatment was associated with fewer beating clusters, a lower proportion of cTnT-positive cells and lower levels of cardiac troponin I (cTnI) and α-myosin heavy chain (α-MHC) expression than those in the control. Conversely, hypoxia treatment was associated with a higher proportion of cTnT-positive cells and higher levels of cTnI expression. In conclusion, transplanted hESCs differentiate toward Flk1-positive cardiac progenitors in the uninjured but not infarcted hearts. The infarcted cardiac microenvironment recapitulated is unsuitable for cardiac differentiation of hESCs, likely due to nutrient deprivation.

Genome-Wide Identification and Characterization of microRNAs in Developing Grains of Zea mays L.

The development and maturation of maize kernel involves meticulous and fine gene regulation at transcriptional and post-transcriptional levels, and miRNAs play important roles during this process. Although a number of miRNAs have been identified in maize seed, the ones involved in the early development of grains and in different lines of maize have not been well studied. Here, we profiled four small RNA libraries, each constructed from groups of immature grains of Zea mays inbred line Chang 7-2 collected 4-6, 7-9, 12-14, and 18-23 days after pollination (DAP). A total of 40 known (containing 111 unique miRNAs) and 162 novel (containing 196 unique miRNA candidates) miRNA families were identified. For conserved and novel miRNAs with over 100 total reads, 44% had higher accumulation before the 9th DAP, especially miR166 family members. 42% of miRNAs had highest accumulation during 12-14 DAP (which is the transition stage from embryogenesis to nutrient storage). Only 14% of miRNAs had higher expression 18-23 DAP. Prediction of potential targets of all miRNAs showed that 165 miRNA families had 377 target genes. For miR164 and miR166, we showed that the transcriptional levels of their target genes were significantly decreased when co-expressed with their cognate miRNA precursors in vivo. Further analysis shows miR159, miR164, miR166, miR171, miR390, miR399, and miR529 families have putative roles in the embryogenesis of maize grain development by participating in transcriptional regulation and morphogenesis, while miR167 and miR528 families participate in metabolism process and stress response during nutrient storage. Our study is the first to present an integrated dynamic expression pattern of miRNAs during maize kernel formation and maturation.

Insulin-producing cells derived from human embryonic stem cells: comparison of definitive endoderm- and nestin-positive progenitor-based differentiation strategies.

Human embryonic stem cells (hESCs) are pluripotent and capable of undergoing multilineage differentiation into highly specialized cells including pancreatic islet cells. Thus, they represent a novel alternative source for targeted therapies and regenerative medicine for diabetes. Significant progress has been made in differentiating hESCs toward pancreatic lineages. One approach is based on the similarities of pancreatic ß cell and neuroepithelial development. Nestin-positive cells are selected as pancreatic ß cell precursors and further differentiated to secrete insulin. The other approach is based on our knowledge of developmental biology in which the differentiation protocol sequentially reproduces the individual steps that are known in normal ß cell ontogenesis during fetal pancreatic development. In the present study, the hESC cell line PKU1.1 was induced to differentiate into insulin-producing cells (IPCs) using both protocols. The differentiation process was dynamically investigated and the similarities and differences between both strategies were explored. Our results show that IPCs can be successfully induced with both differentiation strategies. The resulting IPCs from both protocols shared many similar features with pancreatic islet cells, but not mature, functional ß cells. However, these differently-derived IPC cell types displayed specific morphologies and different expression levels of pancreatic islet development-related markers. These data not only broaden our outlook on hESC differentiation into IPCs, but also extend the full potential of these processes for regenerative medicine in diabetes.

Dynamic expression of microRNAs during the differentiation of human embryonic stem cells into insulin-producing cells.

Human embryonic stem (hES) cells with the capacity of self-renewal and multilineage differentiation are promising sources for generation of pancreatic islet cells for cell replacement therapy in diabetes. Here we induced hES cells into insulin-producing cells (IPCs) in a stepwise process which recapitulated islet organogenesis by directing cells through the stages resembling definitive endoderm, gut-tube endoderm, pancreatic precursor and cells that expressed pancreatic endocrine hormones. The dynamic expression of microRNAs (miRNAs) during the differentiation was analyzed and was compared with that in the development of human pancreatic islets. We found that the dynamic expression patterns of miR-375 and miR-7 were similar to those seen in the development of human fetal pancreas, whereas the dynamic expression of miR-146a and miR-34a showed specific patterns during the differentiation. Furthermore, the expression of Hnf1ß and Pax6, the predicted target genes of miR-375 and miR-7, was reciprocal to that of miR-375 and miR-7. Over-expression of miR-375 down-regulated the expression of gut-endoderm/pancreatic progenitor specific markers Hnf1ß and Sox9. Therefore, the miRNAs may directly or indirectly regulate the expression of pancreatic islet organogenesis-specific transcription factors to control the differentiation and maturation of pancreatic islet cells.

Ghrelin induces cardiac lineage differentiation of human embryonic stem cells through ERK1/2 pathway.

BACKGROUND: Ghrelin, an endogenous ligand for growth hormone secretagogue receptor (GHS-R), shows cardioprotective activity and regulates the differentiation of several mesoderm-derived cells, including myocytes, adipocytes and osteoblasts. The effect of ghrelin on cardiogenesis and its underlying mechanism, however, have not been studied in detail. METHODS: The effects of ghrelin on cardiomyocyte differentiation were tested both in human embryonic stem cells (hESCs) cultured in embryoid body (EB)-based differentiation protocol, and in hESCs transplanted into rat hearts. The signaling mechanisms of ghrelin were further investigated under the EB-based culture condition. RESULTS: The generation of beating EBs and the expression of cardiac-specific markers including cardiac troponin I (cTnI) and α-myosin heavy chain (α-MHC) were 2 to 3-fold upregulated by ghrelin. Although GHS-R1α protein was expressed in differentiated EBs, the effects of exogenous ghrelin were unchanged by D-[lys(3)]-GHRP-6, a specific GHS-R1α antagonist. Moreover, des-acyl ghrelin, which does not bind to GHS-R1α, displayed similar effects with ghrelin. Importantly, activation of ERK1/2, but not Akt, was induced by ghrelin in the newly-formed EBs, and the ghrelin-induced effects of cardiomyocyte differentiation were abolished by adding specific ERK1/2 inhibitor PD98059, but not specific PI3K inhibitor Wortmannin. In addition, ghrelin promoted the differentiation of grafted hESCs into Sox9- and Flk1-positive mesodermal/cardiac progenitor cells in rat hearts. CONCLUSIONS: These results suggest that ghrelin induces cardiomyocyte differentiation from hESCs via the activation of the ERK1/2 signaling pathway. Our study, therefore, indicates that using ghrelin may be an effective strategy to promote the differentiation of hESCs into cardiomyocytes.

Ghrelin promotes the differentiation of human embryonic stem cells in infarcted cardiac microenvironment.

Ghrelin is broadly expressed in myocardial tissues, where it exerts different functions. It also has been found to have a wide variety of biological functions on cell differentiation and tissue development. The aim of this study was to investigate the effect of ghrelin on human embryonic stem cell (hESC) differentiation in infarcted cardiac microenvironment. The hESCs grown on feeder layers expressed several pluripotential markers including alkaline phosphatase (AKP). Four weeks after transplantation into rat infarcted hearts, the hESCs and their progeny cells survived and formed intracardiac grafts were 54.7% and 19.6% respectively in ghrelin- and phosphate-buffered saline (PBS)-treated groups. Double immunostaining with anti-human Sox9 and anti-HNA or anti-human fetal liver kinase-1 (Flk1) and anti ß-tubulin showed that the human grafts were in development. However, double positive stains were only found in the ghrelin-treated group. In addition, the hESC injection protocol was insufficient to restore heart function of the acute myocardial infarction model. Our study, therefore, provides a new insight of ghrelin on promoting hESC survival and differentiation in rat infarcted cardiac microenvironment. This may give a clue for therapy for myocardial infarction by hESCs or progeny cells.

Ghrelin promotes differentiation of human embryonic stem cells into cardiomyocytes.

AIM: Ghrelin is involved in regulating the differentiation of mesoderm-derived precursor cells. The aim of this study was to investigate whether ghrelin modulated the differentiation of human embryonic stem (hES) cells into cardiomyocytes and, if so, whether the effect was mediated by growth hormone secretagogue receptor 1α (GHS-R1α). METHODS: Cardiomyocyte differentiation from hES cells was performed according to an embryoid body (EB)-based protocol. The cumulative percentage of beating EBs was calculated. The expression of cardiac-specific markers including cardiac troponin I (cTnI) and α-myosin heavy chain (α-MHC) was detected using RT-PCR, real-time PCR and Western blot. The dispersed beating EBs were examined using immunofluorescent staining. RESULTS: The percentage of beating EBs and the expression of cTnI were significantly increased after ghrelin (0.1 and 1 nmol/L) added into the differentiation medium. From 6 to 18 d of differentiation, the increased expression of cTnI and α-MHC by ghrelin (1 nmol/L) was time-dependent, and in line with the alteration of the percentages of beating EBs. Furthermore, the dispersed beating EBs were double-positively immunostained with antibodies against cTnI and α-actinin. However, blockage of GHS-R1α with its specific antagonist D-[lys(3)]-GHRP-6 (1 µmol/L) did not alter the effects of ghrelin on cardiomyocyte differentiation. CONCLUSION: Our data show that ghrelin enhances the generation of cardiomyocytes from hES cells, which is not mediated via GHS-R1α.

[Protective effect of calcium dobesilate against early diabetic nephropathy of rat kidney].

The aim of this study is to study the effect of calcium dobesilate on streptozotocin (STZ)-induced early diabetic nephrophathy (DN) in rats. All male Wistar rats were randomly divided into six groups: normal group; DN blank group; calcium dobesilate 75, 150, and 300 mg x kg(-1) groups and perindopril 0.4 mg x kg(-1) group. Blood glucose and the 24 h urinary albumin were measured dynamically during the experiment, after 8 weeks administration, the level of glycosylated hemoglobin (HbA1c) was determined, the expressions of plasminogen activator inhibitor-1 (PAI-1) and matrix metalloprotein-9 (MMP-9) in cortex of kidney were examined with immunohistochemical staining. The endothelin (ET) in plasma and kidney cortex was measured with radioimmunoassay, renal pathomorphism was observed with light and electron microscopes. Calcium dobesilate could decrease the 24 h urinary albumin and ET in plasma and kidney cortex, down-regulate the expression of PAI-1, and up-regulate MMP-9 in kidney. These findings suggested that calcium dobesilate could protect blood vessel endothelium, inhibit kidney fibrous degeneration, ameliorate renal pathological damage, and protect kidney function in many ways.

[Investigation on Fourier transform infrared spectrum of metastatic lymph node in thyroid cancer].

The aim of the study is to explore the possibility of detecting metastatic lymph nodes by FTIR spectra during the surgery of thyroid cancer. The FTIR spectra of 20 metastatic lymph nodes and 69 non-metastatic ones were collected via an ATR (attenuated total reflectance) probe. For each spectrum, 28 variables of 13 bands including peak positions and relative intensities were measured. The variables of metastatic lymph node were compared to those of non-metastatic ones using standard statistic methods. The results indicated that the FTIR spectra of metastatic lymph nodes were significantly different from non-metastatic ones in the bands related to protein, lipid, nucleic acid and carbohydrate. (1) Variations of bands related to protein: The relative intensity ratios of I3 280 /I1 460, I1 640 /I1 460 and I1 546/I1 460 increased significantly (P < 0.05); (2) Variations of bands related to lipid: The relative intensity ratios of I1 743 /I1 460 decreased significantly (P < 0.05). On the contrary, I 400 /I1 460 increased significantly (P < 0.05); (3) The peak positions of 1 165 and 1 120 cm(-1) which were all assigned to carbohydrate shifted toward higher wave number (P < 0.05). The relative intensity ratio of I1 165 /I1 460 decreased significantly (P < 0.05); (4) The peak positions of 1 085 cm(-1) related to nucleic acid shifted to the lower wave number (P < 0.05); (5) Other undetermined bands: The relative intensity ratios of I1 303 /I1 460 and I1 303 /I1 240 increased significantly (P < 0.05). FTIR spectroscopy could be a reliable and practicable method for metastatic lymph nodes diagnosis in the operation of thyroid cancer. It could be applied in detecting metastatic lymph nodes which can not be determined by palpation in surgery.