Association between cigarette smoking and serum alpha klotho levels among US adults over 40-years-old: a cross-sectional study.

Alpha klotho (α-Klotho) is an anti-aging molecule associated with aging and several diseases. Previous studies have reported inconsistent levels of α-Klotho in smokers. This study aimed to demonstrate serum α-Klotho levels in smokers among the US population. This cross-sectional study recruited 11,559 participants (aged 40-79 years; 48.2% males). All data were collected from the 2007-2016 National Health and Nutrition Examination Survey. The study comprised adults with reliable Klotho and smoking questionnaire results. The relationship between smoking and serum α-klotho levels was assessed using multivariate linear regression models after adjusting for potential confounders. We also performed a stratified analysis of clinically important variables. The mean serum α-klotho level among the 11,559 participants was 843.85 pg/mL. After full adjustment, habitual smoking was significantly associated with decreased serum levels of α-klotho level (ß = - 34.89; 95% CI - 54.97, - 14.81; P = 0.0013) in the total study population. Furthermore, the stratified analysis indicated that the association was insignificant in the 60-79 age group. Quitting smoking was not significantly associated with serum levels of α-klotho as expected (P = 0.1148) in the total study population. However, stratified analyses showed a significant inversed association in the male, those with chronic kidney disease, or those with cancer who quit smoking (all P < 0.05). Cigarette smoking was inversely associated with serum α-Klotho levels among US adults.

Association between daily alcohol consumption and serum alpha klotho levels among U.S. adults over 40 years old: a cross-sectional study.

BACKGROUND: Klotho is a hormone considered to be an anti-aging biomarker. The relationships between daily alcohol consumption and serum klotho are mainly unknown. The purpose of this study is to assess the relationship between alcohol consumption and serum alpha klotho (α-klotho) levels in the U.S. METHODS: The data came from 11,558 participants aged ≥ 40 in the 2007-2016 National Health and Nutrition Examination Survey. Adults with reliable α-klotho plasma results were the target population. The self-report method was used to assess alcohol consumption. The relationship between daily alcohol intake and serum α-klotho levels was estimated using multivariable linear regression models. We also performed a stratified analysis of clinically important variables. RESULTS: The mean serum α-klotho level among the 11,558 participants was 843.82 pg/mL. After full adjustment, participants with current moderate and heavy alcohol intake had lower serum α-klotho levels than those who never alcohol intake (ß = - 62.64; 95% CI: - 88.86, - 36.43; P < 0.001; ß = - 81.54; 95% CI: - 111.54, - 51.54; P < 0.001, respectively). Furthermore, the stratified analysis indicated that the association was insignificant in individuals with cardiovascular disease, chronic kidney disease, or cancer. CONCLUSION: Daily alcohol consumption was inversely associated with serum α-klotho levels among U.S. adults over 40 years old. However, individuals with cardiovascular disease, chronic kidney disease, or cancer found no such relationship.

lnc-MRGPRF-6:1 Promotes ox-LDL-Induced Macrophage Ferroptosis via Suppressing GPX4.

Background: Ferroptosis, a newly discovered mode of cell death, emerges as a new target for atherosclerosis (AS). Long noncoding RNAs (lncRNAs) are involved in the regulation of ferroptosis. In our previous study, lnc-MRGPRF-6:1 was highly expressed in patients with coronary atherosclerotic disease (CAD) and closely associated with macrophage-mediated inflammation in AS. In the present study, we aim to investigate the role of lnc-MRGPRF-6:1 in oxidized-low-density lipoprotein (ox-LDL)-induced macrophage ferroptosis in AS. Methods: Firstly, ox-LDL-treated macrophages were used to simulate macrophage injury in AS. Then, ferroptosis-related biomarkers and mitochondrial morphology were detected and observed in ox-LDL-treated macrophages. Subsequently, we constructed lnc-MRGPRF-6:1 knockdown and overexpression of THP-1-derived macrophages and investigated the role of lnc-MRGPRF-6:1 in ox-LDL-induced ferroptosis. Then human monocytes were isolated successfully and were used to explore the role of lnc-MRGPRF-6:1 in macrophage ferroptosis. Likely, we constructed lnc-MRGPRF-6:1 knockdown and overexpression of human monocyte-derived macrophages and detected the expression levels of ferroptosis-related biomarkers. Then, transcriptome sequencing, literature searching, and following quantitative real-time polymerase chain reaction and western blot were implemented to explore specific signaling pathway in the process. It was demonstrated that lnc-MRGPRF-6:1 may regulate ox-LDL-induced macrophage ferroptosis through glutathione peroxidase 4 (GPX4). Eventually, the correlation between lnc-MRGPRF-6:1 and GPX4 was measured in monocyte-derived macrophages of CAD patients and controls. Results: The ox-LDL-induced injury in macrophages was involved in ferroptosis. The knockdown of lnc-MRGPRF-6:1 could alleviate ox-LDL-induced ferroptosis in macrophages. Meanwhile, the overexpression of lnc-MRGPRF-6:1 could intensify ox-LDL-induced ferroptosis. Furthermore, the knockdown of lnc-MRGPRF-6:1 could alleviate the decrease of GPX4 induced by RAS-selective lethal compounds 3 (RSL-3). These indicated that lnc-MRGPRF-6:1 may suppress GPX4 to induce macrophage ferroptosis. Eventually, lnc-MRGPRF-6:1 was highly expressed in the monocyte-derived macrophages of CAD patients and was negatively correlated with the expression of GPX4. Conclusion: lnc-MRGPRF-6:1 can promote ox-LDL-induced macrophage ferroptosis through inhibiting GPX4.

Mechanical properties and immunotherapeutic effects of dissolving microneedles with different drug loadings based on hyaluronic acid

Abstract Improving vaccine immunity and reducing antigen usage are major challenges in the clinical application of vaccines. Microneedles have been proven to be painless, minimally invasive, highly efficient, and have good patient compliance. Compared with traditional transdermal drug delivery, it can effectively deliver a large-molecular-weight drug into the skin, resulting in a corresponding immune response. However, few studies have examined the relationship between microneedle loading dose and immune effects. In this study, the hyaluronic acid (HA) conical and pyramidal dissolving microneedles were prepared by the two-step vacuum drying method, respectively. The model drug ovalbumin (OVA) was added to HA to prepare dissolving microneedles with different loading amounts. The mass ratios of HA to OVA were 5:1, 5:3, and 5:5. The mechanical properties of the dissolving microneedles were characterized using nanoindentation and in vitro puncture studies. The immune effects of the matrix and drug content were studied in Sprague-Dawley (SD) rats. Finally, the diffusion behavior of OVA and the binding mode of HA and OVA in the microneedles were simulated using Materials Studio and Autodocking software. The experimental results showed that the conical microneedles exhibited better mechanical properties. When the mass ratio of HA to OVA was 5:3, the immune effect can be improved by 37.01% compared to subcutaneous injection, and achieved a better immune effect with relatively fewer drugs. This conclusion is consistent with molecular simulations. This study provides theoretical and experimental support for the drug loading and efficacy of microneedles with different drug loadings

Nestin+ Mesenchymal Precursors Generate Distinct Spleen Stromal Cell Subsets and Have Immunomodulatory Function.

Mesenchymal stromal cells (MSCs) are known to be widespread in many tissues and possess a broad spectrum of immunoregulatory properties. They have been used in the treatment of a variety of inflammatory diseases; however, the therapeutic effects are still inconsistent owing to their heterogeneity. Spleen stromal cells have evolved to regulate the immune response at many levels as they are bathed in a complex inflammatory milieu during infection. Therefore, it is unknown whether they have stronger immunomodulatory effects than their counterparts derived from other tissues. Here, using a transgenic mouse model expressing GFP driven by the Nestin (Nes) promoter, Nes-GFP+ cells from bone marrow and spleen were collected. Artificial lymphoid reconstruction in vivo was performed. Cell phenotype, inhibition of T cell inflammatory cytokines, and in vivo therapeutic effects were assessed. We observed Nes-GFP+ cells colocalized with splenic stromal cells and further demonstrated that these Nes-GFP+ cells had the ability to establish ectopic lymphoid-like structures in vivo. Moreover, we showed that the Nes-GFP+ cells possessed the characteristics of MSCs. Spleen-derived Nes-GFP+ cells exhibited greater immunomodulatory ability in vitro and more remarkable therapeutic efficacy in inflammatory diseases, especially inflammatory bowel disease (IBD) than bone marrow-derived Nes-GFP+ cells. Overall, our data showed that Nes-GFP+ cells contributed to subsets of spleen stromal populations and possessed the biological characteristics of MSCs with a stronger immunoregulatory function and therapeutic potential than bone marrow-derived Nes-GFP+ cells.

Synthesis of 2-methoxybenzamide derivatives and evaluation of their hedgehog signaling pathway inhibition.

Aberrant hedgehog (Hh) signaling is implicated in the development of a variety of cancers. Smoothened (Smo) protein is a bottleneck in the Hh signal transduction. The regulation of the Hh signaling pathway to target the Smo receptor is a practical approach for development of anticancer agents. We report herein the design and synthesis of a series of 2-methoxybenzamide derivatives as Hh signaling pathway inhibitors. The pharmacological data demonstrated that compound 21 possessed potent Hh pathway inhibition with a nanomolar IC50 value, and it prevented Shh-induced Smo from entering the primary cilium. Furthermore, mutant Smo was effectively suppressed via compound 21. The in vitro antiproliferative activity of compound 21 against a drug-resistant cell line gave encouraging results.

Development of a UPLC-MS/MS Method for the Quantification of VS-5584 and Its Application in Pharmacokinetic Studies in Rats.

VS-5584 is a small-molecular compound that showed equivalent activity against mTOR and all class I PI3K isoforms and demonstrated preclinical activity in diverse cancer cell lines and xenograft tumor model, and rational combination of VS-5584 and other target therapies achieved promising outcomes in oncology. In the present study, we established and validated a simple and sensitive UPLC-MS/MS method for the determination of VS-5584 in plasma samples. VS-5584 was separated via an Acquity UPLC BEH C18 column, with a mobile phase composed of acetonitrile and 0.2% formic acid in water (40 : 60). The calibration curve displayed a good linearity in the range of 1.0-1000 ng/mL, with satisfactory accuracy (-13.6% < RE% < 8.8%) and precision (CV%, less than 9.2%). The validated method was then applied to a pharmacokinetic study in rats. After administration of 10 mg/kg, VS-5584 was absorbed quickly and reached a peak concentration of 473.2 ± 72.0 ng/mL after 20 min. The established method allows for the quantification of VS-5584 in rat plasma in detail and can be utilized to successfully describe the pharmacokinetic profile of VS-5584.

Development of a ultra-performance LC-MS/MS method for quantification of GW788388 and its applications in pharmacokinetic study in rats.

Aim: GW788388 is a selective and orally active TGF-ß1 receptor inhibitor that shows potent activity in renal fibrosis. We aimed to establish and validate a simple and sensitive ultra-performance LC-MS/MS method for the determination of GW788388 in plasma samples. Methodology & results: GW788388 in rat plasma was processed with protein precipitation method and then separated on a C18 column. The calibration curve presented a good linearity in the range of 1.0-1200 ng/ml, with satisfactory accuracy (relative error, [-17.5% < relative error <11.7%) and precision (CV <8.9%) for all quality control samples. After oral administration, GW788388 was absorbed quickly and reached a peak concentration of 595.3 ± 60.2 ng/ml after 20 min. Conclusion: The validated method provides a quantification method of GW788388 in rat plasma in detail, and can be utilized to successfully describe the pharmacokinetic profile of GW788388.

Tanshinone IIA attenuates demyelination and promotes remyelination in A. cantonensis-infected BALB/c mice.

BACKGROUND: Angiostrongylus cantonensis infection can cause demyelination in the central nervous system, and there is no effective treatment. METHODS: We used dexamethasone, Tanshinone IIA (TSIIA) and Cryptotanshinone(Two traditional Chinese medicine monomers) in combination with albendazole (AB, a standard anti-helminthic compound) to observe their therapeutic effect on demyelination in A. cantonensis-infected mice. Luxol fast blue staining and electron microscope of myelin sheath, Oligodendrocyte (OL) number and myelin basic protein (MBP) expression in brain was detected in above groups. RESULTS: TSIIA+AB facilitated OL proliferation and significantly increased both myelin sheath thickness and the population of small-diameter axons. In addition, TSIIA treatment inhibited the expression of inflammation-related factors (interleukin [IL]-6, IL-1ß, tumor necrosis factor [TNF]-α, inducible nitric oxide synthase [iNOS]) rather than inhibiting eosinophil infiltration in brain. TSIIA also decreased microglial activation and shifted their phenotype from M1 to M2. CONCLUSIONS: Taken together, these results provide evidence that TSIIA combined with AB may be an effective treatment for demyelination caused by A. cantonensis infection and other demyelinating diseases.

Diagnostic value of T1ρ and T2 mapping sequences of 3D fat-suppressed spoiled gradient (FS SPGR-3D) 3.0-T magnetic resonance imaging for osteoarthritis.

Three-dimensional fat-suppressed spoiled gradient magnetic resonance imaging can be used to observe cartilages with high resolution.To quantify and compare the T1ρ and T2 relaxation times of the knee articular cartilage between healthy asymptomatic adults and patients with osteoarthritis (OA).This was a retrospective study of 53 patients with symptomatic OA (6 males and 47 females; aged 57.6 ±â€Š10.0 years) and 26 healthy adults (11 males and 15 females; aged 31.7 ±â€Š12.2 years) from the Ruijin Hospital. T1ρ and T2 relaxation times of knee cartilage were quantified using sagittal multi-echo T1ρ and T2 mapping sequences (3.0-T scanner) and analyzed by receiver operating characteristic (ROC) curve.T1ρ and T2 relaxation times in the OA group were higher than in controls (both P < .01). The sensitivity, specificity, and critical value for differentiating normal from OA cartilage were respectively 92%, 85.6%, and 45.90 ms for T1ρ, and 93.6%, 93.3%, and 50.42 ms for T2. T2 mapping sequence showed a higher area under the ROC curve (AUC) than T1ρ (0.965 vs 0.927, P = .02). The AUC for differentiating normal from Noyes IIA cartilage was 0.922 for T1ρ (cut-off: 46.0; sensitivity: 87.7%; specificity: 89.7%) and 0.954 for T2 (cut-off: 49.5; sensitivity: 91.2%; specificity: 92.3%), with no significant difference between them (P = .08).Both T1ρ and T2 mapping sequences could be used to assess OA cartilage lesions, with T2 mapping sequence demonstrating significant sensitivity for cartilage degeneration. These 2 sequences could also identify early-stage OA cartilage.

Formaldehyde Inhibits Sexual Behavior and Expression of Steroidogenic Enzymes in the Testes of Mice.

BACKGROUND: Formaldehyde, a ubiquitous environmental pollutant, is used extensively and has been proved to impair male reproduction in mammals. However, no trials have explored whether formaldehyde affects sexual function. AIM: To evaluate the effect of long-term formaldehyde exposure on sexual behavior and to investigate the potential mechanism. METHODS: Forty C57BL/6 male mice were randomly allocated to four equally sized groups. Mice were exposed to formaldehyde at a dose of 0 (control), 0.5, 5.0, or 10.0 mg/m3 by inhalation for 60 days. OUTCOMES: Sexual behavior, body and reproductive organ weights, testosterone concentration in serum and testicular tissue, expression of steroidogenic enzymes, quality of sperm, and testicular structure were measured. RESULTS: Formaldehyde inhibited sexual behavior and decreased reproductive organ weights in mice. Serum testosterone levels and intratesticular testosterone concentrations were decreased in the formaldehyde-treated groups. Expression levels of steroidogenic enzymes, including steroidogenic acute regulatory protein, cytochrome P450 cholesterol side-chain cleavage enzyme, and 3ß-hydroxysteroid dehydrogenase (3ß-HSD), also were decreased in the testes of mice exposed to formaldehyde. Moreover, the structure of seminiferous tubules was destroyed and sperm quality decreased after formaldehyde exposure. In addition, the results indicated that the effects of formaldehyde were dose dependent. CLINICAL IMPLICATIONS: Efforts should be undertaken to decrease impairment of sexual function caused by formaldehyde exposure. STRENGTHS AND LIMITATIONS: The relatively small sample might have affected the outcomes. Further experiments are needed to study the mechanism of action of formaldehyde. CONCLUSION: Exposure to formaldehyde gas inhibited sexual behavior, caused reproductive organ atrophy, and impaired spermatogenesis in male mice, which might have been induced by suppressed expression of steroidogenic enzymes in Leydig cells and decreased testosterone synthesis. Zang Z-J, Fang Y-Q, Ji S-Y, et al. Formaldehyde Inhibits Sexual Behavior and Expression of Steroidogenic Enzymes in the Testes of Mice. J Sex Med 2017;14:1297-1306.

Magnetic resonance imaging parameter optimizations for diagnosis of periprosthetic infection and tumor recurrence in artificial joint replacement patients.

To evaluate the efficacy of magnetic resonance imaging (MRI) parameter optimizations for the diagnosis of periprosthetic infection and tumor recurrence in joint replacement patients. We compared the quality of images for 16 joint replacement patients that were recorded using the optimized MRI parameters with and without view angle tilting (VAT) correction at 1.5 T in coronal fast-spin-echo T2-weighted MRI. The optimized MRI data of 86 patients with pain after hip replacement and 67 patients who received tumor resection and joint replacement for bone cancer were retrospectively analyzed to identify MRI features that were useful for the diagnosis of periprosthetic infection and tumor recurrence. Increasing receiver bandwidth and decreasing slice thickness combined with VAT significantly reduced the area of metal-induced artifacts. Irregular soft tissue mass, soft tissue edema, bone destruction, and fistula were significant features of periprosthetic infection, with sensitivities of 47.4-100% and specificities of 73.1-100.0%, which were confirmed based on surgical and pathological findings. Soft tissue mass was a significant feature of tumor recurrence, with 100% sensitivity, 96.0% specificity, and 97.0% consistency. The optimized VAT MRI method demonstrated a high level of diagnostic accuracy for the detection of periprosthetic infection and tumor recurrence in joint replacement patients.

Pentoxifylline Prevents Driamycin-Induced Myocardial Fibrosis and Apoptosis in Rats.

Adriamycin (ADR) is a potent antineoplastic agent, but long-term treatment is limited by its cumulative, life-threatening cardiomyopathy. Recently, a few reports have shown that pentoxifylline (PTX) might produce cardioprotection in cardiac dysfunction. Here, we investigated the protective effects of PTX on ADR-induced cardiomyopathy in rats. Male rats were randomly assigned either to saline, ADR (adriamycin, 5 mg/kg/week), or A (adriamycin, 5 mg/kg/week) + PTX (pentoxifylline, 50 mg/kg/day) groups. After 3 weeks, these animals were sacrificed and the heart tissue was harvested for histological analysis and assessment of hepatocyte growth factor (HGF) and caspase-3 expression. Histopathological findings showed that PTX can alleviate myocardial damage caused by ADR. Cardiac fibrosis was significantly suppressed in the A+PTX group compared to that in the ADR group. The HGF gene expression was decreased significantly in the ADR group compared with the control group, but was increased in the A+PTX group. Caspase-3 was up-regulated in the ADR group, and down-regulated in the A+PTX group. These results show that treatment with PTX exerts a protective effect against ADR-induced myocardial fibrosis via regulation of HGF and caspase-3 gene expression. PTX may thus represent a useful new clinical tool for the treatment of ADR-induced cardiomyopathy.

Engraftable neural crest stem cells derived from cynomolgus monkey embryonic stem cells.

Neural crest stem cells (NCSCs), a population of multipotent cells that migrate extensively and give rise to diverse derivatives, including peripheral and enteric neurons and glia, craniofacial cartilage and bone, melanocytes and smooth muscle, have great potential for regenerative medicine. Non-human primates provide optimal models for the development of stem cell therapies. Here, we describe the first derivation of NCSCs from cynomolgus monkey embryonic stem cells (CmESCs) at the neural rosette stage. CmESC-derived neurospheres replated on polyornithine/laminin-coated dishes migrated onto the substrate and showed characteristic expression of NCSC markers, including Sox10, AP2α, Slug, Nestin, p75, and HNK1. CmNCSCs were capable of propagating in an undifferentiated state in vitro as adherent or suspension cultures, and could be subsequently induced to differentiate towards peripheral nervous system lineages (peripheral sympathetic neurons, sensory neurons, and Schwann cells) and mesenchymal lineages (osteoblasts, adipocytes, chondrocytes, and smooth muscle cells). CmNCSCs transplanted into developing chick embryos or fetal brains of cynomolgus macaques survived, migrated, and differentiated into progeny consistent with a neural crest identity. Our studies demonstrate that CmNCSCs offer a new tool for investigating neural crest development and neural crest-associated human disease and suggest that this non-human primate model may facilitate tissue engineering and regenerative medicine efforts.

Bioinformatics analysis of Rab GDP dissociation inhibitor beta and its expression in non-small cell lung cancer.

BACKGROUND: Lung cancer has been considered as one of the most important causes of cancer-related mortality worldwide. To predict lung cancer, researchers identified several molecular markers. However, many underlying markers of lung cancer remain unclear. One of these markers is Rab GDP dissociation inhibitor beta (GDIß), which is related to tumorigenicity, development and invasion. This study was designed to analyze the biological characteristics of Rab GDIß and to detect the mRNA and protein expressions of Rab GDIß in lung cancer cells; this study also aimed to investigate the functions of this protein in lung cancer. METHOD: Using online software from the websites of NCBI, ProtParam and so on, we analyzed the biological characteristics of Rab GDIß. RT-PCR was performed to detect gene expressions in A549 and 16HBE cell lines and immunohistochemistry (IHC) staining was conducted to detect Rab GDIß protein expression in 57 cases of human lung cancer tissues and 19 cases of normal lung tissues. The association of protein expression with patient clinical and pathological characteristics was assessed in each dataset. RESULTS: Bioinformatic analysis on Rab GDIß: The mRNA of human Rab GDIß contains two transcript variants; the common structural elements of the two proteins are mainly α-helix, random coil, ß-turn and extended strand. Three and four transmembrane domains could be found in the entire polypeptide chain of protein variants 1 and 2, respectively; both transcript variants are hydrophilic and soluble proteins. The RT-PCR result: The mRNA expression of Rab GDIß was down-regulation in A549 cells compared with that in 16HBE cells. The IHC result: The protein expression of Rab GDIß in lung cancer cells was significantly lower than that in normal lung tissues (P <0.05) but was not correlated with patients' age, gender, tumor size, pathological type, differentiation, lymph node metastasis, distant metastasis and TNM stage. CONCLUSION: The expression of Rab GDIß was low in non-small cell lung cancer (NSCLC). Hence, Rab GDIß may be a tumor suppressor and could function as an indicator of tumorigenesis in NSCLC; nevertheless, this result should be further studied. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_201.

Clinical value of optimized magnetic resonance imaging for evaluation of patients with painful hip arthroplasty.

BACKGROUND: The imaging evaluation of pain in patients who have had a hip arthroplasty (HA) is challenging, and traditional imaging techniques, including magnetic resonance imaging (MRI) and computerized tomography (CT), are limited by metallic artifact. The purpose of the present study was to investigate the use of modified MRI techniques to visualize periprosthetic soft tissues and the bone-implant interface, and to evaluate the value of MRI for the assessment of patients with painful hip arthroplasty. METHODS: Fifty-six painful hips in fifty-six patients following primary HA were assessed using optimized MRI, CT and standardized radiographs. The diagnosis of MRI was correlated with intraoperative findings as well as with microbiological and histological examinations (when available). The sensitivity and the specificity of MRI diagnosis were determined according to final diagnosis. The chi-square test was performed to detect a difference between MRI and final diagnosis. RESULTS: Forty-eight patients have received revision surgery and final diagnosis were established. MRI was demonstrated high sensitivity and specificity in detecting aseptic loosening (93% and 95%), periprosthetic infection (94% and 97%), adverse local tissue reaction (100% and 100%) and periprosthetic fracture (100% and 100%). MRI was determined to be the most sensitive technique in detecting implant loosening for any reason, with a sensitivity of 93.8% for acetabular shell and 97.1% for femoral stem, compared to 81.3% and 80.0% on CT, 75.0% and 77.1% on radiographs. CONCLUSIONS: Optimized MRI was effective for the assessment of the periprosthetic soft tissues and bone. The use of modified magnetic resonance imaging parameters provided a useful adjunct to conventional examinations for the evaluation of patients with painful hip arthroplasty.

Bioinformatics analysis and expression study of fumarate hydratase in lung cancer.

BACKGROUND: As its etiology and pathogenesis is obscure, illustrating the molecular mechanism of lung cancer has become a serious and urgent task. Studies have shown that fumarate hydratase (FH) is a tumor suppressor related to tumorigenesis, development, and invasion. Our aim was to analyze the biological information of FH, and detect the messenger ribonucleic acid (mRNA) and protein expression of FH in lung cancer cells to explore its role in tumorigenesis and in the development of lung cancer. METHOD: We analyzed the biological characteristics of FH, then utilized reverse transcription-polymerase chain reaction (RT-PCR) to study FH mRNA expression in A549 and 16 human bronchial epithelial (HBE) cell lines. The protein expression of FH was detected in 57 cases of human lung cancer tissues and 19 cases of normal lung tissues by immunohistochemistry. RESULTS: 1. Bioinformatic analysis: FH mainly exist in the mitochondria; the common structural elements of FH are mainly α-helix, random coil, ß-turn, and extended strand; there are five possible transmembrane domains in the entire polypeptide chain; FH is a hydrophilic and soluble protein. 2. RT-PCR result: FH mRNA expression was downregulated in A549 cells compared with 16HBE cells. 3. Immunohistochemistry: FH protein expression was significantly lower in lung cancer cells than in normal lung tissues (P < 0.05), but was not correlated with the patients' age, gender, tumor size, pathological type, or lymph node, distant, or tumor node metastasis stage. CONCLUSION: FH was under-expressed in lung cancer, suggesting that it may be an indicator of tumorigenesis and could be a potential target for therapies against lung cancer in the future.

S-adenosylmethionine-induced adipogenesis is accompanied by suppression of Wnt/ß-catenin and Hedgehog signaling pathways.

S-Adenosylmethionine (SAM) plays a crucial role as a methyl donor in various biological processes and has been previously shown to be involved in adipogenesis in skeletal muscle. This study was conducted to explore the mechanism of SAM inducing adipogenesis in skeletal muscle. Adipose precursor cells, 3T3-L1, and C2C12 cells, were induced into adipogenic differentiation by addition of SAM in MDI-differentiation media (0.5 mmol/L isobutylmethylxanthine, 1 µm/L dexamethasone, and 10 µg/mL insulin) to explore the role of SAM in promoting adipogenesis. Subsequently, cells were cultured with a medium containing SAM alone at the beginning of differentiation to test the relationship between SAM-induced adipogenesis and Wnt/ß-catenin, and Hedgehog signaling pathways that control the cell commitment to adipogenic- or myogenic-differentiation. We found SAM possessed an additive effect with MDI in promoting adipogenesis of 3T3-L1 and C2C12 cells at the beginning of adipogenic differentiation. SAM could also individually induce cell adipogenesis in a dose-dependent manner. Moreover, the expression of Wnt/ß-catenin and Hedgehog signals and their targets were suppressed by SAM (P < 0.05). These results demonstrate that SAM, as an increasingly accepted nutritional supplement, can initiate adipogenesis of adipose precursor cells derived from adipose and muscle tissues, a function at least partly correlated with the suppression of Wnt/ß-catenin and Hedgehog pathways.

[Expression of P75NTR in the testis of nestin-GFP transgenic mice].

OBJECTIVE: To explore the P75NTR expression in the mouse testis and its relationship with nestin. METHODS: We observed the location of the expressions of P75NTR and nestin in the testis of the nestin-GFP transgenic mouse on postnatal day (PND) 5, 14 and 30 using immunofluorescence, and detected the expression levels of P75NTR in the testicular tissue of mice in different age groups by real-time quantitative PCR (RTqPCR) and flow cytometry. Then we cultured the P75NTR positive cells in neural stem cell culture medium and observed their neuronal differentiation capacity by orientation differentiation. RESULTS: Immunofluorescence showed the expressions of P75NTR and nestin in the Leydig cells of the mouse testis. RTqPCR and flow cytometry exhibited the peak of the P75NTR expression on PND 14. The positive rates of P75NTR were (2.88 +/- 0.52), (9.54 +/- 1.81) and (2.63 +/- 0.43)% on PND 5, 14 and 30, respectively. The P75NTR positive cells obtained also expressed nestin and P75NTR and had the capacity of neuronal differentiation. CONCLUSION: P75NTR and nestin are co-expressed in the Leydig cells of the mouse testis, and the P75NTR positive cells have the ability of neural differentiation, which is presumably attributed to neural crest cells.

Suicide gene-mediated ablation of tumor-initiating mouse pluripotent stem cells.

Pluripotent stem cells, including embryonic stem (ES) and induced pluripotent stem (iPS) cells, serve as unlimited resources for cell replacement therapy and tissue engineering because such cells are capable of extensive proliferation in vitro and can give rise to lineages that represent any of the three embryonic germ layers. However, in the context of the in vivo behavior of cell transplants, key challenges need to be addressed and essential strategies should be developed before stem cells can be used in clinical practice. In the present study, we modified mouse ES/iPS cells to contain a suicide gene, deltaTK or CodA, under the transcriptional control of the EF1α or Nanog promoter. The suicide gene was introduced via lentivirus transduction without interfering with their self-renewal and pluripotency characteristics. We found that EF1α promoter-controlled deltaTK/CodA expression efficiently eliminated pluripotent stem cells and their derivatives both in vitro and in vivo. When the suicide gene was under the control of the Nanog promoter, tumor-initiating undifferentiated pluripotent stem cells were selectively ablated in vitro after prodrug treatment. These results indicate that modification of pluripotent stem cells with a suicide gene prior to transplantation offers a safe manner by which wayward stem cells, and their progeny, can be controlled in vivo. Our approach will render the clinical application of human pluripotent stem cells increasingly possible.