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The aim of this study is to evaluate the diagnostic value of genome sequencing in children with epilepsy, and to provide genome sequencing-based insights into the molecular genetic mechanisms of epilepsy to help establish accurate diagnoses, design appropriate treatments and assist in genetic counselling. We performed genome sequencing on 320 Chinese children with epilepsy, and interpreted single-nucleotide variants and copy number variants of all samples. The complete pedigree and clinical data of the probands were established and followed up. The clinical phenotypes, treatments, prognoses and genotypes of the patients were analysed. Age at seizure onset ranged from 1 day to 17 years, with a median of 4.3 years. Pathogenic/likely pathogenic variants were found in 117 of the 320 children (36.6%), of whom 93 (29.1%) had single-nucleotide variants, 22 (6.9%) had copy number variants and two had both single-nucleotide variants and copy number variants. Single-nucleotide variants were most frequently found in SCN1A (10/95, 10.5%), which is associated with Dravet syndrome, followed by PRRT2 (8/95, 8.4%), which is associated with benign familial infantile epilepsy, and TSC2 (7/95, 7.4%), which is associated with tuberous sclerosis. Among the copy number variants, there were three with a length, Zou et al. perform whole genome sequencing in 320 Chinese children with epilepsy, and investigate single nucleotide variants and copy number variants in all individuals. The results confirm the value of whole genome sequencing as a first-tier diagnostic test for patients with epilepsy.

Tuberous sclerosis complex (TSC) is a multi-system genetic disorder. Most patients have germline mutations in TSC1 or TSC2 but, 10%-15% patients do not have TSC1/TSC2 mutations detected on routine clinical genetic testing. We investigated the contribution of low-level mosaic TSC1/TSC2 mutations in unsolved sporadic patients and families with TSC. Thirty-one sporadic TSC patients negative on routine testing and eight families with suspected parental mosaicism were sequenced using deep panel sequencing followed by droplet digital polymerase chain reaction. Pathogenic variants were found in 22/31 (71%) unsolved sporadic patients, 16 were mosaic (median variant allele fraction [VAF] 6.8% in blood) and 6 had missed germline mutations. Parental mosaicism was detected in 5/8 families (median VAF 1% in blood). Clinical testing laboratories typically only report pathogenic variants with allele fractions above 10%. Our findings highlight the critical need to change laboratory practice by implementing higher sensitivity assays to improve diagnostic yield, inform patient management and guide reproductive counseling.

The malar augmentation injection has gained popularity in recent years, but the exact location of each injection site has not been clearly identified.To discover ideal injection sites by comprehensively considering the distributions of ligaments, muscles, and vessels.Eighteen cadaver heads were dissected to investigate the zygomatic ligamentous system and to measure the position of muscles. Sixty-six cadaver heads were subjected to computed tomographic scanning and three-dimensional vessel reconstruction. Radiological evaluation of the fillers was performed before and after experimental injection in one hemiface and dissected to confirm safe delivery. Five patients were enrolled in a prospective clinical study. 2D and 3D photographs were taken before and after the injections for comparison.Site 1 was defined along the zygomatic arch, except the first 1/4 length and the midline of the arch. Site 2 was on the body of the zygoma, superior to the level of the infraorbital foramen and medial to the jugale. Site 3 was defined in the anteromedial midface approximately 30 mm below the lateral canthus.Injections at these 3 sites can be performed within the range of the ligaments to achieve effective lifting effects and minimize potential complications.

Background Strengthening weakened ligament tissues with injectable fillers to improve their supportive effect may achieve the aesthetic goal of face lifting. Objectives The aim of the study was to design an injectable technique for enhancing the true facial ligaments and dissect the ligaments to provide anatomical guidance for effective injection. Methods Six true facial ligaments were chosen as target anatomical sites for injection. Specimens were dissected, and 3-dimensional (3D) images were reconstructed to confirm the exact location of each injection site and to confirm that the proposed injection routes will not cause dangerous vascular damage. A total of 5 patients received the injections; 3D images were taken before and after the injections for comparison and clinical outcome assessments. Results The injection technique was designed to target 6 true facial ligaments, as follows. Site 1 targeted the temporal ligamentous adhesion region to lift the lateral ends of the eyebrows. Site 2 targeted the region of the lateral orbital thickening to lift the lateral canthus. Site 3 and site 4 targeted the zygomatic retaining ligaments and zygomatic cutaneous ligaments, respectively, to augment the soft tissues of the midface. Site 5 targeted the region of the maxillary ligament to lessen the nasolabial folds, and site 6 targeted the mandibular ligament to reduce the marionette line. Conclusions This site-specific injection technique targeting the true ligaments may lead to increased efficiency and accuracy of face rejuvenation and exert a lifting effect. Level of Evidence: 4

Ellagic acid (EA), a natural polyphenol compound that exists in a variety of fruits and vegetables, has been reported to inhibit tumor growth by reducing cell growth, inducing apoptosis, and damaging mitochondria. Recent reports demonstrate that mitochondria regulate cancer cell death through energy metabolism and that different types of cell death coexist in vivo. We showed that EA inhibited lung cancer cell proliferation, markedly decreased ATP levels, decreased the potential of the inner mitochondrial membrane and decreased oxygen consumption in vitro. In addition, EA activated AMP-activated protein kinase (AMPK) and reduced HIF-1α in lung cancer cells. Moreover, the treatment of tumor-bearing mice with EA dramatically inhibited tumor growth, increased p-AMPK and suppressed HIF-1α levels. These findings suggest that EA could be a promising chemotherapeutic agent that targets mitochondrial metabolism in lung cancer.

Anatomical knowledge of the zygomatic cutaneous ligament is crucial for rejuvenation of the anteromedial midface. However, there is a lack of satisfactory descriptions of the anatomy of the zygomatic cutaneous ligament, and the exact range and location are still controversial. The present study attempts to clarify the anatomy of the zygomatic cutaneous ligament to provide vital information for clinical operations.Facial dissection was performed on 36 cadaver hemifaces. The location of the zygomatic cutaneous ligament was investigated and recorded relative to the Frankfort horizontal line and several vertical reference lines. The relative relationship of the zygomatic cutaneous ligament with surrounding anatomical structures was also investigated.The zygomatic cutaneous ligament is a septum-like osteocutaneous ligament originating from the periosteum of the maxilla and zygoma. The overall range of the zygomatic cutaneous ligament starts at the origin of the levator labii superioris and then extends laterally, following the curvature of the inferior bone margin. After merging with the ligamentous part at the origin of zygomaticus minor and zygomaticus major muscle (11.65 mm inferior to the horizontal line), it continues as the zygomatic retaining ligament on the zygomatic arch. The vertical distances between the zygomatic cutaneous ligament and horizontal line along the L1, L2, L3, L4, and L5 reference lines are 9.1, 19.5, 22.1, 21.7, and 18.7 mm, respectively.The anatomical data obtained in the present study regarding the location of the zygomatic cutaneous ligament might be valuable for understanding the appearance of the midcheek fold and be helpful for surgical procedures to rejuvenate the anteromedial midface.

Background Descending necrotizing mediastinitis (DNM) is one of the most virulent forms of mediastinitis. The main causes of high mortality in DNM are believed to stem from difficulty and delay in the diagnosis. Fast and accurate identification of pathogens is important for the treatment of these patients. Metagenomics next-generation sequencing (mNGS) is a powerful tool to identify all kinds of pathogens, especially for rare and complex infections. Case presentation A 64-year-old male patient was admitted to the intensive care unit (ICU) with unconsciousness, dyspnea, and swelling in the mandible and neck. Computed tomography (CT) scan results combined with clinical laboratory examination indicated DNM. Vancomycin and imipenem were used, and vacuum sealing drainage was applied for debridement and drainage of the infected area. The positive mNGS results of drainage fluid confirmed the presence of mixed infection caused by Streptococcus anginosus, Prevotella oris, and several other anaerobes. The antibiotics were adjusted to piperacillin/tazobactam and tinidazole according to the mNGS results and antimicrobial susceptibility testing of cultured pathogens. After 11 days of antibiotic therapy, the infection symptoms of the neck and mediastinum improved, and the patient was transferred out of the ICU on the 26th day after negative result of drainage fluid culture. Conclusion This case suggested that mNGS is a promising technology for precise and fast pathogens identification with high sensitivity, which may guide the diagnosis of infectious diseases in the future trend.

Chemical investigation on the aerial part of Achillea alpina L. led to the isolation of twenty sesquiterpenoids. The structures of the undescribed achigermalides A-H were determined by extensive spectroscopic analysis, including NMR, HRESIMS, UV and IR, and their absolute configurations were established by computational electronic circular dichroism (ECD) method. The X-ray crystal structure for 8α-angeloxy-1β,2β:4β,5β-diepoxy-10β-hydroxy-6βH,7αH,11βH-12,6α-guaianolide was reported for the first time. Glucose consumption was analyzed to investigate the effect of all compounds on palmitic acid (PA)-mediated insulin resistance (IR) in HepG2 cells, and achigermalides D-F, desacetylherbohde A, and 4E,10E-3-(2-methylbutyroyloxy)-germacra-4,10(1)-diene-12,6α-olide appreciably enhanced the glucose consumption at low concentrations of 1.56-6.25 μM. Moreover, achigermalide D decreased the expression of IL-1β and the generation of reactive oxygen species (ROS), and also down-regulated the protein levels of TXNIP, NLRP3, caspase-1 and NF-κB in the Western blot analysis, suggesting achigermalide D mediated IR via the suppression of NLRP3 inflammasome pathway.

Enhanced secondary aerosol formation was observed during the COVID-19 lockdown in Xi'an, especially for polluted episodes. More oxidized‑oxygenated organic aerosol (MO-OOA) and sulfate showed the dominant enhancements, especially in large particle-mode. Meanwhile, relative humidity (RH) showed a positive promotion on the formation of sulfate and MO-OOA during the lockdown, but had no obvious correlation with less oxidized‑oxygenated organic aerosol (LO-OOA) or nitrate. Organosulfurs (OS) displayed a higher contribution (~58%) than inorganic sulfate to total sulfate enhancement in the polluted episode during the lockdown. Although the total nitrate (TN) decreased during the lockdown ascribing to a larger reduction of inorganic nitrate, organic nitrate (ON) showed an obvious increase from pre-lockdown (0.5 ± 0.6 μg m

Worship activities like burning joss paper during the Chinese Hanyi festival is a common, traditional custom in northwest China. However, the pollutants of e.g., soot particles, released from joss paper burning and the corresponding impacts on urban air quality were poorly investigated, which can be a particular concern since these activities are conducted in an uncontrolled manner. In this study, a long time-of-flight (LToF) soot particle aerosol mass spectrometry (SP-AMS) was deployed to characterize the refractory black carbon (rBC) emitted from the joss paper burning, as well as crop residue, coal combustion, and traffic during the Hanyi Festival in mid-November 2020 in the northwestern city of Xi'an in China. Large difference (from5% to100%) in the fragmentation patterns (C

The COVID-19 pandemic has drastically affected the economic and social activities, leading to large reductions in anthropogenic emissions on a global scale. Despite the reduction of primary emissions during the lockdown period, heavy haze pollution was observed unexpectedly in megacities in North and East China. In this study, we conducted online measurements of organic aerosol in a background site before and during the lockdown in Guanzhong basin, Northwest China. The oxygenated organic aerosol (OOA) increased from 24% of total OA (3.2 ± 1.6 μg m−3) before lockdown to 54% of total OA (4.5 ± 1.3 μg m−3) during lockdown, likely due to substantial decrease of NOx emissions during lockdown which resulted in large increase of O3 and thus atmospheric oxidizing capacity. OOA showed higher mass concentrations and fractional contributions during lockdown than before lockdown, and increased with the increase of Ox in both periods. In comparison, aqueous secondary organic aerosol (aqSOA) showed high mass concentrations and fractional contributions in both polluted periods before and during lockdown with the increase of aerosol liquid water content (ALWC). The increase of aqSOA under high ALWC conditions is very likely the reason of pollution events during lockdown. Combined with trajectory analysis, the absence of Guanzhong cluster in polluted period during lockdown may play a key role in the OA variations between two polluted periods. In addition, when comparing the clusters from the same transmission directions between before lockdown and during lockdown, the OA fractions showed similar variations during lockdown in all clusters, suggesting the OA variations are widespread in northwest China., Graphical abstract Unlabelled Image

Objective This study investigated the frequency of T-helper (Th)17 lymphocytes and production of cytokine interleukin (IL)-17 in peripheral blood of patients with non-small-cell lung cancer (NSCLC) and their use as a marker of clinical value. Methods Sixty patients with NSCLC and 60 healthy volunteers were enrolled in the study. Flow cytometry was used to detect the frequency of Th17 lymphocytes in peripheral blood, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of IL-17. We analyzed the association of Th17 lymphocytes and IL-17 levels in the peripheral blood of patients with their clinicopathological features. Results Frequency of Th17 lymphocytes and production of IL-17 were significantly higher in the NSCLC group than in the control group and were higher in patients with a smoking history compared with non-smokers. Moreover, Th17 lymphocyte and IL-17 expression levels were higher in patients with squamous cell carcinoma than in patients with adenocarcinoma, and significantly higher in patients with stage III and IV cancers than in patients at stage I or II. Conclusion Th17 lymphocytes and IL-17 play an important role in the development of NSCLC in patients and may have clinical value as markers for treatment of NSCLC.

Background The current study aims to evaluate the correlation between peripheral blood miR-148a-3p level in patients with liver injury after hepatectomy under general anesthesia with propofol, thereby evaluating whether circulating miR-148a-3p can be used as a biomarker of liver injury after hepatectomy. Methods One hundred one patients who underwent hepatectomy at the Fourth Hospital of Xi'an were selected. On the first day after operation, peripheral blood was taken to determine the level of miR-148a-3p in peripheral blood. The correlation between circulating miR-148a-3p and liver injury after operation was analyzed using Pearson's correlation assay. Results First, our data showed that the level of miR-148a-3p was increased in the peripheral blood of patients after hepatectomy. The amount of bleeding was significantly correlated with the increase of ALT, AST, and miR-148a-3p. Moreover, the level of miR-148a-3p was significantly correlated with alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Further analysis showed that intraoperative propofol dosage was correlated with ALT, AST, and miR-148a-3p after hepatectomy. Conclusions MiR-148a-3p may be sensitive to ischemic and traumatic liver injury and may be a marker of liver injury after hepatectomy under general anesthesia with propofol.

Background Ovarian cancer is one of the most common malignant tumors in the female reproductive system with the highest mortality rate. Cul4B participates in the oncogenesis and progression of several malignant tumors. However, the role of Cul4B in ovarian cancer has not been studied. Results High expression of intratumor Cul4B was associated with poor patient survival. Cul4B expression was associated with FIGO stage and Cul4B was independent risk factor of ovarian cancer disease-free survival and overall survival. In vitro studies revealed that overexpression of Cul4B promoted tumor proliferation while knockdown of Cul4B significantly inhibited the proliferation capacity of ovarian cancer cells. Mechanistically, Cul4B was found to promotes cell entering S phase from G0/G1 phase by regulating the expression of CDK2 and CyclinD1. Cul4B regulates the expression of CDK2 and CyclinD1 by repressing miR-372. Conclusions The results revealed that high expression of Cul4B is associated with poor ovarian cancer prognosis and Cul4B may serve as a potential treating target for an adjuvant therapy.

The presence of organochlorine pesticides (OCPs) in dairy products can lead to human exposure. This study investigated the behavior of OCP residues in milk during yogurt and cheese production. Gas chromatography with electron-capture detection (GC-ECD) was used to detect α-hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), γ-HCH, g-chlordane, and α-chlordane in fresh milk, yogurt, and cheese. The results showed that fermentation reduced the residual concentration of OCPs in yogurt, with processing factors (PFs) ranging from 0.42 to 0.64. The reductions in residue levels during fermentation were due to the activity of the starter. The cheese making process increased the residual concentration of OCPs in cheese compared to raw milk, with PFs ranging from 2.37 to 4.93. Additionally, milk, yogurt, and cheese samples were purchased from local markets and OCP levels were analyzed. The target OCPs ranged from ND to 16.50 μg/kg in these samples.

Humic-like substances (HULIS) are ubiquitous in the atmospheric environment, which affects both human health and climate. We present here the mass concentration and optical characteristics of HULIS isolated from aerosol samples collected in Xi'an, China. Both mass concentration and absorption coefficient (Abs365) of HULIS show clear seasonal differences, with the highest average in winter (3.91 μgC m-3 and 4.78 M m-1, respectively) and the lowest in summer (0.65 μgC m-3 and 0.55 M m-1, respectively). The sources of HULIS_C and light absorption of HULIS were analyzed by positive matrix factorization (PMF) and four major sources were resolved, including secondary formation, biomass burning, coal burning, and vehicle emission. Our results show that secondary formation (i.e., gas-to-particle conversion from e.g., photochemical oxidation) was the major contributor to both HULIS_C (50%) and light absorption (55%) of HULIS in summer, biomass burning and coal burning were major sources of HULIS_C (~70%) and light absorption (~80%) of HULIS in winter. It is worth noting that biomass burning and coal burning had higher contribution to HULIS light absorption (47% in spring, 37% in summer, 73% in fall, and 77% in winter) than their corresponding contribution to HULIS_C concentration (41% in spring, 37% in summer, 54% in fall, and 69% in winter). However, vehicle emission had lower contribution to HULIS light absorption (26% in spring, 8% in summer, 18% in fall, and 11% in winter) than to HULIS_C concentration (24% in spring, 13% in summer, 28% in fall, and 18% in winter). These results suggest that HULIS from biomass burning and coal burning have higher light absorption ability than from vehicle emission.

Background Chronic brain hypoperfusion (CBH) is closely related to Alzheimer’s disease (AD) and vascular dementia (VaD). Meanwhile, synaptic pathology plays a prominent role in the initial stage of AD and VaD. However, whether and how CBH impairs presynaptic plasticity is currently unclear. Methods In the present study, we performed a battery of techniques, including primary neuronal culture, patch clamp, stereotaxic injection of the lentiviral vectors, morris water maze (MWM), dual luciferase reporter assay, FM1–43 fluorescence dye evaluation, qRT-PCR and western blot, to investigate the regulatory effect of miR-153 on hippocampal synaptic vesicle release both in vivo and in vitro. The CBH rat model was generated by bilateral common carotid artery ligation (2VO). Results Compared to sham rats, 2VO rats presented decreased field excitatory postsynaptic potential (fEPSP) amplitude and increased paired-pulse ratios (PPRs) in the CA3-CA1 pathway, as well as significantly decreased expression of multiple vesicle fusion-related proteins, including SNAP-25, VAMP-2, syntaxin-1A and synaptotagmin-1, in the hippocampi. The levels of microRNA-153 (miR-153) were upregulated in the hippocampi of rats following 2VO surgery, and in the plasma of dementia patients. The expression of the vesicle fusion-related proteins affected by 2VO was inhibited by miR-153, elevated by miR-153 inhibition, and unchanged by binding-site mutation or miR masks. FM1–43 fluorescence images showed that miR-153 blunted vesicle exocytosis, but this effect was prevented by either 2′-O-methyl antisense oligoribonucleotides to miR-153 (AMO-153) and miR-masking of the miR-153 binding site in the 3′ untranslated region (3’UTR) of the Snap25, Vamp2, Stx1a and Syt1 genes. Overexpression of miR-153 by lentiviral vector-mediated miR-153 mimics (lenti-pre-miR-153) decreased the fEPSP amplitude and elevated the PPR in the rat hippocampus, whereas overexpression of the antisense molecule (lenti-AMO-153) reversed these changes triggered by 2VO. Furthermore, lenti-AMO-153 attenuated the cognitive decline of 2VO rats. Conclusions Overexpression of miR-153 controls CBH-induced presynaptic vesicle release impairment by posttranscriptionally regulating the expression of four vesicle release-related proteins by targeting the 3’UTRs of the Stx1a, Snap25, Vamp2 and Syt1 genes. These findings identify a novel mechanism of presynaptic plasticity impairment during CBH, which may be a new drug target for prevention or treatment of AD and VaD. Graphical abstract

Cdc42, a member of the Rho GTPases family, is involved in the regulation of several cellular functions including cell cycle progression, survival, transcription, actin cytoskeleton organization and membrane trafficking. Diabetes is a chronic and metabolic disease, characterized as glycometabolism disorder induced by insulin deficiency related to β cell dysfunction and peripheral insulin resistance (IR). Diabetes could cause many complications including diabetic nephropathy (DN), diabetic retinopathy and diabetic foot. Furthermore, hyperglycemia can promote tumor progression and increase the risk of malignant cancers. In this review, we summarized the regulation of Cdc42 in insulin secretion and diabetes-associated diseases. Organized researches indicate that Cdc42 is a crucial member during the progression of diabetes, and Cdc42 not only participates in the process of insulin synthesis but also regulates the insulin granule mobilization and cell membrane exocytosis via activating a series of downstream factors. Besides, several studies have demonstrated Cdc42 as participating in the pathogenesis of IR and DN and even contributing to promote cancer cell proliferation, survival, invasion, migration, and metastasis under hyperglycemia. Through the current review, we hope to cast light on the mechanism of Cdc42 in diabetes and associated diseases and provide new ideas for clinical diagnosis, treatment, and prevention.

The chemical and cytotoxicity properties of fine particulate matter (PM

Background The link between cardiac diseases and cognitive deterioration has been accepted from the concept of “cardiogenic dementia”, which was proposed in the late 1970s. However, the molecular mechanism is unclarified. Methods The two animal models used in this study were cardiac-specific overexpression of microRNA-1-2 transgenic (Tg) mice and a myocardial infarction mouse model generated by left coronary artery ligation (LCA). First, we observed the microRNA-1 (miR-1) level and synaptic vesicles (SV) distribution in the hippocampus using in situ hybridization and transmission electron microscopy (TEM) and evaluated the expression of vesicle exocytosis related proteins by western blotting. Second, we used dual luciferase reporter assay as well as antagonist and miRNA-masking techniques to identify the posttranscriptional regulatory effect of miR-1 on the Snap25 gene. Third, FM1–43 staining was performed to investigate the effect of miR-1 on synaptic vesicle exocytosis. Lastly, we used GW4869 to inhibit the biogenesis and secretion of exosomes to determine the transportation effect of exosomes for miR-1 from the heart to the brain. Results Compared with the levels in age-matched WT mice, miR-1 levels were increased in both the hearts and hippocampi of Tg mice, accompanied by the redistribution of SVs and the reduction in SV exocytosis-related protein SNAP-25 expression. In vitro studies showed that SNAP-25 protein expression was down- or upregulated by miR-1 overexpression or inhibition, respectively, however, unchanged by miRNA-masking the 3’UTR of the Snap25 gene. SV exocytosis was inhibited by miR-1 overexpression, which could be prevented by co-transfection with an anti-miR-1 oligonucleotide fragment (AMO-1). The knockdown of miR-1 by hippocampal stereotaxic injection of AMO-1 carried by a lentivirus vector (lenti-pre-AMO-1) led to the upregulation of SNAP-25 expression and prevented SV concentration in the synapses in the hippocampi of Tg mice. The application of GW4869 significantly reversed the increased miR-1 level in the blood and hippocampi as well as reduced the SNAP-25 protein levels in the hippocampi of both Tg and LCA mice. Conclusion The overexpression of miR-1 in the heart attenuated SV exocytosis in the hippocampus by posttranscriptionally regulating SNAP-25 through the transportation of exosomes. This study contributes to the understanding of the relationship between cardiovascular disease and brain dysfunction.

The contamination of polycyclic aromatic hydrocarbons (PAHs) in surface dusts attracts great attentions due to their properties of threatening human health. Twenty-nine surface dust samples were collected from driving-schools in a city of Henan. Concentrations of 16 priority PAHs were analyzed by gas chromatography-mass spectrometry (GC-MS). The health risks exposed to dust PAHs for three different scenarios (working for 5 a, 10 a and 20 a in driving-school) were estimated by the health risk assessment model (ILCRs). Source identification was analyzed by diagnostic ratio, composition analysis, and principal component analysis. The results showed that concentrations of the ∑PAHs in dusts ranged from 198.21 to 3400.89 μg·kg

IL-13 is a proinflammatory cytokine associated with multiple pathological conditions and the promotion of metastasis in lung cancer. Previous studies have demonstrated that IL-13 and YY1 are associated with PI3K/AKT signaling. In addition, miR-29a has been found to play a critical role in cell invasion in lung cancer. However, the molecular mechanism of miR-29a underlying its involvement in IL-13-induced lung cancer cell invasion remains largely unknown. In the present study, we aimed to investigate the role of miR-29a in cell invasion mediated by IL-13 in lung cancer. By using MTT and wound-scratch assays, we assessed cell proliferation and migration induced by IL-13, and identified activation of the PI3K/AKT/YY1 pathway. Inhibition of PI3K/AKT by LY294002 downregulated IL-13-induced YY1 expression. Furthermore, we found that miR-29a directly targets YY1 and suppressed its expression in lung cancer. By using MTT, flow cytometry and Transwell assays, overexpression of miR-29a restricted both YY1 and N-cadherin expression, and inhibited IL-13-induced invasion of lung cancer A549 cells. Taken together, these findings demonstrate that PI3K/AKT/YY1 is involved in the regulation of lung cancer cell behavior induced by IL-13, and miR-29a represents a promising therapeutic target.

Background aims Since human embryonic stem cells and human fetal neural stem cells have immune rejection and ethical issues, recent advancements in induced pluripotent stem cells (iPS cells) provide new possibilities to study autologous cell therapy for Parkinson's disease (PD). Methods We isolated human skin fibroblasts from normal individuals and patients with PD; we generated iPS cells by transfecting these human skin fibroblasts with retroviral reprogramming factors of OCT4, SOX2, KLF4 and c-MYC and induced iPS cells to differentiate neural stem cells (NSCs) and then into neurons and dopamine neurons in vitro . Results We found that iPS cell–derived NSC transplant into the striatum of the 6-hydroxydopamine (OHDA)–induced PD rats improved their functional defects of rotational asymmetry at 4, 8, 12 and 16 weeks after transplantation. iPS cell–derived NSCs were found to survive and integrate into the brain of transplanted PD rats and differentiated into neurons, including dopamine neurons in vivo . Conclusions Transplantation of iPS cell–derived NSCs has therapeutic potential for PD. Our study provided experimental proof for future clinical application of iPS cells in cell-based treatment of PD.

Anticancer activity and toxicity of doxorubicin (Dox) are associated with its DNA intercalation. To understand the role in gene regulation and the drug mechanism, it is a challenge to detect the DNA-Dox interaction at the single-molecule level without the use of laborious, time-consuming labeling assays and an error-prone amplification method. Here, we utilized the simplest and cheapest, yet highly sensitive, single-molecule nanopore technology to investigate the DNA-Dox interaction and explore in situ the intercalative reaction kinetics. Distinctive electronic signal patterns between DNA and the DNA-Dox complex allow protein nanopore to readily detect the changes in structure and function of DNA. After Dox insertion, nanopore unzipping time of DNA was elevated 10-fold while the blocking current decreased, demonstrating the higher affinity of the DNA-Dox complex (formation constant K(f) = 3.09 × 10(5) M(-1)). Continuous rapid nanopore detection in real time displayed that Dox intercalation in DNA is a two-state dynamic process: fast binding and slow conformational adaption. The nanopore platform provides a powerful tool for studying small molecule-biomacromolecule interactions and paves the way for novel applications aimed at drug screening and functional analysis.

Background This study explored the neural differentiation and therapeutic effects of stem cells from human exfoliated deciduous teeth (SHED) in a rat model of Parkinson's disease (PD). Methods The SHED were isolated from fresh dental pulp and were induced to differentiate to neurons and dopamine neurons by inhibiting similar mothers against dpp (SMAD) signaling with Noggin and increase conversion of dopamine neurons from SHED with CHIR99021, Sonic Hedgehog (SHH) and FGF8 in vitro. The neural-primed SHED were transplanted to the striatum of 6-hydroxydopamine (6-OHDA)–induced PD rats to evaluate their neural differentiation and functions in vivo. Results These SHED were efficiently differentiated to neurons (62.7%) and dopamine neurons (42.3%) through a newly developed method. After transplantation, the neural-induced SHED significantly improved recovery of the motor deficits of the PD rats. The grafted SHED were differentiated into neurons (61%), including dopamine neurons (22.3%), and integrated into the host rat brain by forming synaptic connections. Patch clamp analysis showed that neurons derived from grafted SHED have the same membrane potential profile as dopamine neurons, indicating these cells are dopamine neuron-like cells. The potential molecular mechanism of SHED transplantation in alleviating motor deficits of the rats is likely to be mediated by neuronal replacement and immune-modulation as we detected the transplanted dopamine neurons and released immune cytokines from SHED. Conclusion Using neural-primed SHED to treat PD showed significant restorations of motor deficits in 6-OHDA–induced rats. These observations provide further evidence that SHED can be used for cell-based therapy of PD.

Supranutritional selenite has anti-cancer therapeutic effects in vivo; however, the detailed mechanisms underlying these effects are not clearly understood. Further studies would broaden our understanding of the anti-cancer effects of this compound and provide a theoretical basis for its clinical application. In this study, we primarily found that selenite exposure inhibited phosphorylation of cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB), leading to suppression of Bcl-2 in HCT116 and SW480 colorectal cancer (CRC) cells. Moreover, the selenite-induced inhibitory effect on PKD1 activation was involved in suppression of the CREB signalling pathway. Additionally, we discovered that selenite treatment can upregulate p38 MAPK phosphorylation, which results in inhibition of the PKD1/CREB/Bcl-2 survival pathway and triggers apoptosis. Finally, we established a colorectal cancer xenograft model and found that selenite treatment markedly inhibits tumour growth through the MAPK/PKD1/CREB/Bcl-2 pathway in vivo. Our results demonstrated that a supranutritional dose of selenite induced CRC cell apoptosis through inhibition of the PKD1/CREB/Bcl-2 axis both in vitro and in vivo.

The Wnt signaling pathway is well known to play major roles in skeletal development and homeostasis. In certain aspects, fracture repair mimics the process of bone embryonic development. Thus, the importance of Wnt signaling in fracture healing has become more apparent in recent years. Here, we summarize recent research progress in the area, which may be conducive to the development of Wnt-based therapeutic strategies for bone repair. [BMB Reports 2014; 47(12): 666-672]

Duchenne and Becker muscular dystrophies (DMD/BMD) are the most commonly inherited neuromuscular disease. However, accurate and convenient molecular diagnosis cannot be achieved easily because of the enormous size of the dystrophin gene and complex causative mutation spectrum. Such traditional methods as multiplex ligation-dependent probe amplification plus Sanger sequencing require multiple steps to fulfill the diagnosis of DMD/BMD. Here, we introduce a new single-step method for the genetic analysis of DMD patients and female carriers in real clinical settings and demonstrate the validation of its accuracy. A total of 89 patients, 18 female carriers and 245 non-DMD patients were evaluated using our targeted NGS approaches. Compared with traditional methods, our new method yielded 99.99% specificity and 98.96% sensitivity for copy number variations detection and 100% accuracy for the identification of single-nucleotide variation mutations. Additionally, this method is able to detect partial deletions/duplications, thus offering precise personal DMD gene information for gene therapy. We detected novel partial deletions of exons in nine samples for which the breakpoints were located within exonic regions. The results proved that our new method is suitable for routine clinical practice, with shorter turnaround time, higher accuracy, and better insight into comprehensive genetic information (detailed breakpoints) for ensuing gene therapy.

Oral squamous cell carcinoma (OSCC) is currently a highly prevalent disease worldwide. Cisplatin (CDDP) is widely used for the chemotherapy of OSCC. Yet, the molecular mechanisms responsible for cisplatin resistance have not been fully elucidated. In this study, we showed that overexpression of p21 (RAC1) activated kinase 1 (PAK1) induced epithelial to mesenchymal transition (EMT) and significantly promoted the invasion and migration of oral squamous cell carcinoma SCC25 cells. Emerging evidence indicates a strong link between resistance to therapy and the induction of EMT in cancer. We showed that overexpression of PAK1 induced cisplatin resistance in SCC25 cells. ERCC1 and YAP can promote cisplatin resistance in human OSCC. We showed that ERCC1 and YAP protein were upregulated by PAK1 in SCC25 cells. -We found that miR‑485‑5p inhibited PAK1 protein expression in the SCC25 cells. Contrary to PAK1, we demonstrated that overexpression of miR‑485‑5p reversed EMT and significantly inhibited invasion and migration. Moreover, its overexpression sensitized SCC25-CR cells (cisplatin-resistant cells) to cisplatin. Thus, we conclude that miR‑485‑5p reverses EMT and promotes cisplatin-induced cell death by targeting PAK1 in oral tongue squamous cell carcinoma. This study suggests that PAK1 plays an essential role in the progression of OSCC and it is a potential therapeutic target for OSCC.

To explore the relationships between pathogenic gene mutations and extra-gonadal manifestations in male patients with congenital hypogonadotropic hypogonadism (CHH).A total of 15 genes were screened in 259 CHH patients by massively parallel DNA sequencing. The identified pathogenic mutations were confirmed by Sanger polymerase chain reaction (PCR). The extra-gonadal features in all patients were recorded.Gene mutations were detected in 83 (32.0%) patients, which included FGFR1 (n=20), PROKR2 (n=19), CHD7 (n=16), KAL1 (n=14) and other genes (n=14), such as PROK2, FGF8, GNRHR, KISS1R, NELF and WDR11. Patients with KAL1 mutation displayed CHH related family history (n=5) and obesity (n=3). Patients with FGFR1 mutation presented with short stature (n=3), testicular hydrocele (n=2), lip-palate cleft (n=2) and dental dysplasia (n=1). Patients without detectable mutations may also exhibit obesity, mental retardation and short stature.CHH patients may present with extra-gonadal manifestations, including anosmia, obesity, dental dysplasia, short stature and mental retardation. An obscure relationships could be found between the clinical features and different gene mutations, but it is difficult to speculate the types of pathogenic gene mutations according to these extra-gonadal manifestations.

High-dose methotrexate (HDMTX) chemotherapy is generally accepted as an effective method for the treatment and prevention of extramedullary leukemia in children. However, it is unknown whether HDMTX chemotherapy kills intestinal bacteria on a large scale, thus causing dysbacteriosis, which may in turn influence the progress or prognosis of leukemia. The aim of this study was to examine changes in intestinal flora in children with acute lymphoblastic leukemia (ALL) treated with HDMTX chemotherapy. Bacterial DNA in stool from 36 healthy children and 36 ALL children were tested at A260 with a spectrophotometer before and after HDMTX chemotherapy. The primers of Bifidobacteria, Lactobacillus and Escherichia coli were designed according to the 16SrRNA/DNA bacterial sequences. Bacteria were qualitatively and quantitatively confirmed by routine polymerase chain reaction (PCR) and fluorescent quantitative PCR, respectively. Our data showed that the total amount of flora in the stools of children with ALL was decreased by 29.6% compared with healthy children ( P < 0.01). The total amount of flora in the stools of children with ALL on the third and seventh days after chemotherapy were 1496.5 ± 577.1 and 1966.6 ± 598.3 ng/ μL, respectively, which was notably less than before chemotherapy (2436.3 ± 768.6 ng/ μL). The amount of Bifidobacteria, Lactobacillus and E. coli in the intestinal tract in the ALL group after chemotherapy had an apparent change, which decreased most clearly on the third day, and partially recovered on the seventh day after chemotherapy. HDMTX chemotherapy can cause intestinal dysbacteriosis in children with ALL. The amount of Bifidobacteria, Lactobacillus and E. coli decreased significantly compared with the control group.

A novel strategy for the enhancement of electrochemiluminescence (ECL) was developed by combining CdS quantum dots (QDs), graphene (G) and agarose. This enhanced ECL was exploited to develop a label-free ECL immunosensor for the ultrasensitive detection of alpha fetoprotein (AFP). The novel G–CdS QDs–agarose composite was first coated on the glass carbon electrode surface to form a robust film, which exhibited high ECL intensity, good biocompatibility and high stability. After that 3-aminopropyl-triethoxysilane (APS), as a binding linker, was conjugated to the G–CdS QDs–agarose composite film on the electrode, the ECL signal was significantly enhanced. The fabrication of ECL immunosensor was successfully completed by immobilizing the AFP-antibody (Ab) onto the electrode through glutaric dialdehyde (GLD). The specific immunoreaction between AFP and antibody resulted in the decrease in ECL intensity and the intensity decreased linearly with the logarithm of AFP concentration in the range of 0.0005–50 pg mL−1 with a detection limit of 0.2 fg mL−1. The immunosensor exhibits high sensitivity, specificity, stability, reproducibility and good regeneration, thus has the potential to be used in clinical application. Besides, the highly enhanced ECL from the G–CdS QDs–agarose composite film opened new avenues to apply graphene and QDs ECL in analytical systems and ECL biosensors.

Despite great improvements in the diagnosis and treatment of neoplasms, metastatic disease is still the leading cause of death in cancer patients, with mortality rates still rising. Given this background, new ways to treat cancer will be important for development of improved cancer control strategies. Cdc42 is a member of the Rho GTPase family and plays an important role in cell-to-cell adhesion, formation of cytoskeletal structures, and cell cycle regulation. It thus influences cellular proliferation, transformation, and homeostasis, as well as the cellular migration and invasion processes underlying tumor formation. Cdc42 acts as a collection point for signal transduction and regulates multiple signaling pathways. Moreover, recent studies show that in most human cancers Cdc42 is abnormally expressed and promoting neoplastic growth and metastasis. Regarding possible new treatments for cancer, miRNA and small molecules targeting Cdc42 and related pathways have been recently found to be effective on cancer. In this review, we analyze the newly recognized regulation mechanisms for Cdc42 and Cdc42-related signal pathways, and particularly new treatments using small molecules and miRNAs to inhibit the abnormal overexpression of Cdc42 that may slow down the metastasis process, improve cancer therapy and lead to novel strategies for development of antineoplastic drugs.

Summary Objective Estrogen deficiency results in postmenopausal osteoporosis by increasing the rate of bone loss. The mechanism responsible for the effects of estrogen on osteoclasts is still unclear. Materials and Methods The potential of mononuclear cells from cord blood or bone marrow to differentiate into mature osteoclasts when co-cultured with human osteoblast cells was investigated. The effects of estrogen on osteoclastogenesis and osteoclast activity were also examined. Results Macrophage markers CD11b and CD14 were downregulated and vitronectin receptor was upregulated during 28 days' co-culture of mononuclear cells and human osteoblasts. Long-term co-culture resulted in the formation of numerous large tartrate-resistant acid phosphatase-positive multinucleated cells capable of resorption of bone slices. After incubation for 28 days, the addition of 17β-estradiol caused a significant decrease in the expression of vitronectin receptor and tartrate-resistant acid phosphatase-positive multinucleated cells in cultures derived from both bone marrow and cord blood. A significant decrease in bone resorption was also noted in the presence of estrogen. Conclusion Estrogen not only suppresses osteoclastogenesis but also inhibits the activity of osteoclasts.

Aim: The objectives of this study were to investigate the inhibitory action of verapamil on wild-type(WT) and mutation HERG K + channel current (I HERG ), and to determine whether mutations in the S6 region are important for the inhibition of I HERG by verapamil. Methods: HERG channels (WT, Y652A, and F656A) were expressed in oocytes of Xenopus laevis and studied using the 2-electrode voltage-clamp technique. Results: WT HERG is blocked in a concentration-dependent manner by verapamil (half-maximal inhibition concentration [IC 50 ]=5.1 μmol/L), and the steady state activation and inactivation parameters are shifted to more negative values. However, mutation to Ala of Y652 and F656 located on the S6 domain produced 16-fold and 20-fold increases in IC 50 for I HERG blockade, respectively. Simultaneously, the steady state activation and inactivation parameters for Y652A are also shifted to more negative values in the presence of the blockers. Conclusion: Verapamil preferentially binds to and blocks open HERG channels. Tyr-652 and Phe-656, 2 aromatic amino-acid residues in the inner (S6) helix, are critical in the verapamil-binding site.

Background From 1973 to 2005, the lung cancer mortality in Xuanwei had increased constantly. Effect analysis of age and non-age factors on lung cancer is important for local policy-making. Methods Demographic and death data was collected and used. Factors of lung cancer were classified into age and non-age factors. The contribution of the two factors to lung cancer was evaluated by method of decomposing the differences of mortality rate. Results For males, the non-age factors were the major contributor to growth of lung cancer mortality, and 78.46 % of all growth was attributed to non-age factors. For females, the non-age factors were the absolute contributor to growth of lung cancer in 1973–1992. From 1992 to 2005, the contribution proportion had reduced to 75.39 %. Conclusions Aging was one of risk factors for lung cancer in Xuanwei, but not the main factor. It was supposed that multiple environmental risk factors were related with high growth of lung cancer in Xuanwei. Policy-making should focus on the non-age factors.

Ingestion of contaminated soil has been recognized as an important exposure pathway of cadmium (Cd) for humans, especially for children through outdoor hand-to-mouth activities. The effect of ageing process following the input of Cd into soil on the bioaccessibility of Cd in five typical soils of China was investigated using physiologically based in vitro test in this study. A sequential extraction procedure was employed with attempt to identify the bioaccessible fraction(s) of Cd in soils. The bioaccessibility of Cd in strongly acidic (≈pH 4.5) soils reached nearly steady levels (76.5–76.9% and 52.0–52.6% in the gastric and small intestinal phases, respectively) after a sharp decline in the first week of ageing. In contrast, the bioaccessibility of Cd in higher pH (>6.0) soils was found to be much lower (53.3–72.7% and 29.9–43.4% in gastric and small intestinal phases, respectively) and took 2 weeks of ageing to reach steady levels. The freshly spiked Cd was more labile than native Cd. The main proportion of spiked Cd was found in exchangeable Cd which was higher in strongly acidic soils (68.6–71.8%) than in higher pH soils (53.4–61.4%) at day 120 after a sharp decline to the nearly steady state in the first 1 and 2 weeks, respectively. Significant correlations between Cd bioaccessibility and either water soluble and exchangeable Cd individually, or the sum of water soluble and exchangeable Cd throughout the incubation period for all soils, indicate that these forms of Cd are likely to constitute the main proportion of bioaccessible Cd in soils. Keywords: Soil, Cadmium, Ageing, Bioaccessibility, Sequential extraction procedure

As an important human exposure pathway of contaminants, soil ingestion is of increasing concern for assessing health risk from polycyclic aromatic hydrocarbons (PAHs) in soils. A wide range of total PAH concentrations ranging from 0.112 microg g(-1) to 27.8 microg g(-1) in soils collected from different public sites, including gas stations, roadsides, bus stops, a kindergarten, primary and middle schools, a university and residential area, was detected. In general, total PAHs concentrations in soils from traffic areas were significantly higher than that from the other sites, indicating a dominant contribution from motor vehicles. Physiologically based in vitro tests were used to evaluate the oral bioaccessibility of PAHs in surface soil under different land uses in Beijing regarding both gastric and small intestinal conditions. It was found that the oral bioaccessibility of total PAHs in small intestinal condition, ranging from 9.2% to 60.5% of total PAHs in soil, was significantly higher than gastric condition, ranging from 3.9% to 54.9%. The bioaccessibility of individual PAHs in soils generally decreased with the increasing ring number of PAHs in both gastric and small intestinal conditions. However, the ratio of bioaccessibility of individual PAHs in gastric condition to that in small intestinal condition, generally increased with increasing ring number, indicating the relatively pronounced effect of bile extract on improving bioaccessibility of PAHs with relatively high ring numbers characterized by their high K(ow) values. The observation that bile extract at a level higher than critical micelle concentration could reduce the surface tension of digestive juice substantially, which may cause PAHs to be available for intestinal absorption, calls for more careful establishment of reliable soil criteria for PAHs, especially concerning the health of children who may ingest a considerable amount of PAH-contaminated soil via outdoor hand-mouth activities.

To investigate the prevalence of childhood asthma, and to find the distribution characteristics, precipitating factors, diagnosis and treatment status, and to provide scientific data for improving the prevention and management of asthma in children in Kunming City, China.Children were selected by random cluster sampling. A standardized preliminary questionnaire was used for screening out possible patients in the survey. Diagnosis of asthma was confirmed by diagnostic criteria in suspected asthmatic children. Asthmatic children were further asked for past diagnosis and treatment with the questionnaire of asthma in children.The total asthma incidence rate was 1.40%. The prevalence of asthma in male and female children was 1.89% and 0.88% respectively (P0.05). Children aged 0-5 years old had a higher prevalence of asthma (1.69%) than that of school-age children (6-14 years old, 1.21%). In all asthmatic children, 51.3% were previously diagnosed with classical asthma or cough variant asthma, 26.0% were suffered attacks from December to February, and 54.0% were suffered attacks at midnight or dawn. Respiratory tract infection (87.3%) was the most common triggers of asthma exacerbation. Antibiotics were used in 80.0%, bronchodilators in 66.0%, inhaled corticosteroid in 64.0%. A peak flow meter for monitoring lung function was used in 17% of asthmatic children over 5 years old.The prevalence of asthma is associated with age and gender in children aged 0-14 years old in Kunming City. Acute asthma attack occurs mostly in winter and at midnight or dawn. Respiratory tract infection is the most common trigger of asthma exacerbation. Nearly a half of patients with asthma had not been diagnosed with asthma in the early stage. Most asthmatic children use antibiotics and only two-thirds use bronchodilators or inhaled corticosteroid in the treatment. The treatment and management of asthma in children awaits improvement as well.

Severe retinal ischemia causes persistent visual impairments in eye diseases. Retinal pigment epithelium (RPE) cells are located near the choroidal capillaries, and are easily affected by ischemic or hypoxia. Ginsenoside Rg-1 has shown significant neuroprotective effects. This study was performed to test the cytoprotective effect of ginsenoside Rg-1 in RPE cells against hypoxia and cobalt chloride (CoCl2) assaults, and to understand the underlying mechanisms. We found that Rg-1 pre-administration significantly inhibited CoCl2- and hypoxia-induced RPE cell death and apoptosis. Reactive oxygen specisis (ROS)-dependent p38 and c-Jun NH(2)-terminal kinases (JNK) MAPK activation was required for CoCl2-induced RPE cell death, and Rg-1 pre-treatment significantly inhibited ROS production and following p38/JNK activation. Further, CoCl2 suppressed pro-survival mTOR complex 1 (mTORC1) activation in RPE cells through activating of AMP-activated protein kinase (AMPK), while Rg-1 restored mTORC1 activity through inhibiting AMPK activation. CoCl2-induced AMPK activation was also dependent on ROS production, and anti-oxidant N-acetylcysteine (NAC) prevented AMPK activation and RPE cell death by CoCl2. Our results indicated that Rg-1 could be further investigated as a novel cell-protective agent for retinal ischemia.

To investigate the effects of cis-combretastatin-A1 phosphate (cis-CA1P) on tumor cell proliferation, and its effects on the blood vessel formations.MTT and IC50 values were used to assess the inhibitory effects of cis-CA1P on tumor cell proliferation. Chicken embryo chorioallantoic membrane and thoracic aorta annulations isolated from rats were used to investigate the effects of cis-CAIP on the blood vessel formation.Cis-CA1P concentration-dependently inhibited the proliferations of several cancer cell lines, including human gastric carcinoma cell line MGC-803, human leukemic monocyte lymphoma cell line U937, human melanoma cell line A375, human colon cancer cell line HCT116, human breast carcinoma cell line MDA-MB-231, and human leukemia cell line K562. Cis-CAIP significantly decreased the formation of blood vessels in chicken embryo chorioallantoic membrane and in thoracic aorta annulations.Cis-CA1P inhibits cancer cell proliferation and prevents blood vessel formation.

To study the pulmonary pathology in patients died of fatal human influenza A(H1N1) infection.Eight cases of fatal human influenza A (H1N1) infection, including 2 autopsy cases and 6 paramortem needle puncture biopsies, were enrolled into the study. Histologic examination, immunohistochemitry, flow cytometry and Western blotting were carried out.The major pathologic changes included necrotizing bronchiolitis with surrounding inflammation, diffuse alveolar damage and pulmonary hemorrhage. Influenza viral antigen expression was detected in the lung tissue by Western blotting. Immunohistochemical study demonstrated the presence of nuclear protein and hemagglutinin virus antigens in parts of trachea, bronchial epithelium and glands, alveolar epithelium, macrophages and endothelium. Flow cytometry showed that the apoptotic rate of type II pneumocytes (32.15%, 78.15%) was significantly higher than that of the controls (1.93%, 3.77%).Necrotizing bronchiolitis, diffuse alveolar damage and pulmonary hemorrhage followed by pulmonary fibrosis in late stage are the major pathologic changes in fatal human influenza A (H1N1) infection.

Tissue engineering provides a new paradigm for periodontal tissue regeneration in which proper stem cells and effective cellular factors are very important. The objective of this study was, for the first time, to investigate the capabilities and advantages of periodontal tissue regeneration using induced pluripotent stem (iPS) cells and enamel matrix derivatives (EMD). In this study the effect of EMD gel on iPS cells in vitro was first determined, and then tissue engineering technique was performed to repair periodontal defects in three groups: silk scaffold only; silk scaffold + EMD; and silk scaffold + EMD + iPS cells. EMD greatly enhanced the mRNA expression of Runx2 but inhibited the mRNA expression of OC and mineralization nodule formation in vitro. Transplantation of iPS cells showed higher expression levels of OC, Osx, and Runx2 genes, both 12 and 24 days postsurgery. At 24 days postsurgery in the iPS cell group, histological analysis showed much more new alveolar bone and cementum formation with regenerated periodontal ligament between them. The results showed the commitment role that EMD contributes in mesenchymal progenitors to early cells in the osteogenic lineage. iPS cells combined with EMD provide a valuable tool for periodontal tissue engineering, by promoting the formation of new cementum, alveolar bone, and normal periodontal ligament.

A gas chromatography-mass spectrometry assay was developed and validated for the simultaneous determination of phthalates and adipates in human serum. The phthalates and adipates studied were dimethyl phthalate, diethyl phthalate, dibutyl phthalate, benzylbutyl phthalate, di-2-ethylhexyl phthalate, di-n-octyl phthalate, diethyl adipate, dibutyl adipate, diisobutyl adipate, bis(2-butoxyethyl) adipate and di-2-ethylhexyl adipate, with diisooctyl phthalate as internal standard. The extraction and cleaning up procedure was carried out with solid-phase extraction cartridges containing dimethyl butylamine groups, which showed extraction efficiencies over 88% for each analyte and the internal standard. The calibration curves obtained were linear with correlation coefficients greater than 0.98. For all analytes, the assay gave CV% values for intra-day precision from 4.9 to 13.3% and mean accuracy values from 91.4 to 108.4%, while inter-day precision was 5.2-13.4% and mean accuracy 91.0-110.2%. The limits of detection for the assay of phthalates and adipates were in the range 0.7-4.5 ng/mL. The method is simple, sensitive and accurate, and allows for simultaneous determination of nanogram levels of phthalates and adipates in human serum. It was successfully applied to an investigation on the level of phthalates and adipates in a non-occupationally exposed population.

To identify the possible microRNAs (miRNAs) which target the polycystic kidney disease-2 gene (PKD2), and clarify effects of the miRNAs on PKD2. We preliminarily used bioinformatics to analyze 3′UTR (3′untranslated regions) of PKD1 and PKD2 in order to predict the potential microRNAs targeted on them. Subsequently, the stable cell lines with overexpression of microRNA-17 (miR-17) were screened, and luciferase assay combined with the mutation 3′UTR of PKD2 were performed to verify PKD2 is the target of miR-17. Moreover, RT-PCR and Western Blotting were used to determine the post-transcriptionally regulation of PKD2 by miR-17. Finally, MTT cell assays allied with PKD2 rescued strategy were employed to evaluate cell proliferation effects. Our study firstly found that the 3′UTR of PKD2 was more conservation than that of PKD1, and microRNA-17 directly targets the 3′UTR of PKD2 and post-transcriptionally repress the expression of PKD2. Moreover, our findings also demonstrated that overexpression of miR-17 may promote cell proliferation via post-transcriptionally repression of PKD2 in HEK 293T. This suggested that microRNA might be a novel mechanism for cystogenesis as well as a potential therapeutic target for the cell proliferation of autosomal dominant polycystic kidney disease (ADPKD).

To investigate the effects of high-dose methotrexate (HDMTX) therapy on intestinal bacterial flora in children with acute lymphoblastic leukemia (ALL).Thirty-six children with ALL of pre-and post-HDMTX therapy and 36 control children were enrolled. The bacterial DNA in stool was extracted. The primers for Bacillus bifidus and Escherichia coli with the 16SrRNA/DNA sequence of bacteria were designed. The bacteria were identified through general PCR. The standard curve of both bacterial DNA was produced using a series of dilution of accurately quantified bacterial DNA. The unknown samples were measured by 16SrRNA/DNA-targeted PCR. The amount of stool Bacillus bifidus and Escherichia coli among 36 control children and 36 children with ALL of pre- and post-HDMTX therapy were obtained by using the standard curves.Bacillus bifidus logarithmic absolute value of the first day before treatment, of third day after treatment, of seventh day after treatment in patients with ALL and the control was 7.24 +/- 0.33, 6.00 +/- 0.27, 6.59 +/- 0.33, and 9.49 +/- 0.41, respectively (P0.01). Escherichia coli logarithmic absolute value of the first day before treatment, of third day after treatment, of seventh day after treatment in patients with ALL and the control was 6.62 +/- 0.42, 5.96 +/- 0.42, 7.02 +/- 0.41, and 7.52 +/- 0.43, respectively (P0.01). The logarithm of the amount of stool Bacillus bifidus and Escherichia coli in control group was higher in ALL group (F = 739.61, 88.67, P0.01). There were significant difference (P0.01) in the logarithm of the amount of stool Bacillus bifidus and Escherichia coli between pre-therapy and post-therapy group. The logarithm of the bacterium was very low on third day after treatment, but gradually increased on the seventh day after treatment.(1) HDMTX therapy has great effects on intestinal flora of Bacillus bifidus and Escherichia coli and significantly reduced the bacteria in children with ALL. (2) Probiotics had significantly decreased in ALL group than in control group.

Ingestion of contaminated soil has been recognized as an important exposure pathway of arsenic for humans, especially for children through outdoor hand-to-mouth activities. An improved sequential extraction procedure was employed in an attempt to reveal the relationship between bioaccessibility and fractionation of As in five soils from China. Arsenic bioaccessibility in acidic ( approximately pH 4.5) soils reached approximately stable levels after a sharp decline within one week of ageing. In contrast, As bioaccessibility in higher pH (6.0) soils was found to be significantly higher and took two weeks of ageing to reach stable levels. The artificially added As was more labile than indigenous As. The main proportions of added As were found in the specifically sorbed and amorphous and poorly-crystalline hydrous Fe/Al oxide-bound fractions. Correlation analysis shows that the non-specifically and specifically sorbed As are likely to constitute the main proportion of bioaccessible soil As. The soil content of amorphous and crystalline Fe/Al oxides and soil pH appear to be the key factors controlling, not only the time needed to reach a steady state, but also the magnitude of the bioaccessibility of As added to the soils.

Neuroendocrine testing was carried out in seven male volunteers before and at the end of a 10-day course of carbamazepine (up to 700 mg daily). After carbamazepine treatment, the prolactin response to intravenous administration of the serotonin precursor tryptophan (5 g) was significantly enhanced, but there was no change in basal plasma tryptophan level or in tryptophan disposition after infusion. The prolactin response to intravenous protirelin (6.25 micrograms) was unaltered. Carbamazepine treatment also produced an increase in the growth hormone response to subcutaneous administration of the dopamine agonist apomorphine hydrochloride (5 micrograms/kg). These data suggest that carbamazepine may alter brain serotonin and dopamine function in humans. Such effects could be involved in the therapeutic properties of carbamazepine in affective disorder.