Gene expression profiles in the brain of phenylketonuria mouse model reversed by the low phenylalanine diet therapy.

To gain insight into the potential protective mechanisms of low phenylalanine diet (LPD) in phenylketonuria (PKU), gene expression profiles were studied in the cerebral cortex and hippocampus of a PKU mouse model (BTBR-Pahenu2). PKU mice were fed with low Phe diet (LPD-PKU group) and normal diet (PKU group). Wild-type mice were treated with normal diet (WT group) as control. After 12 weeks, we detected gene expression in the cerebral cortex and hippocampus of the three groups by RNA-sequencing, and then screened the differentially-expressed genes (DEGs) among the groups by bioinformatics analyses. We found that the transcriptional profiles of both cerebral cortex and hippocampus changed markedly between PKU and WT mice. Furthermore, LPD changed the transcriptional profiles of the cerebral cortex and the hippocampus of PKU mice significantly, especially in the cerebral cortex, with overlaps of genes that changed with the disease and altered by LPD treatment. In the cerebral cortex, hundreds of DEGs enriched in a wide spectrum of biological processes, molecular function, and cellular component, including nervous system development, axon development and guidance, calcium ion binding, modulation of chemical synaptic transmission, and regulation of protein kinase activity. In the hippocampus, the overlapping genes were enriched in positive regulation of long term synaptic, negative regulation of excitatory postsynaptic potential, positive regulation of synapse assembly. Our results showed that genes impaired in PKU and then rescued by LPD might indicate the potential protective capability of LPD in the PKU brain.

Therapeutic Effects of Punicalagin Against Ovarian Carcinoma Cells in Association With ß-Catenin Signaling Inhibition.

AIM: The aim of this study was to investigate the effects of punicalagin, a polyphenol isolated from Punica granatum, on human A2780 ovarian cancer cells in vitro. METHODS: The viability of human A2780 ovarian cells was evaluated using Cell Counting Kit-8 assay. Cell cycle was detected with flow cytometry analysis. The protein expression levels of Bcl-2, Bax, ß-catenin, cyclin D1, survivin, tissue inhibitor of metalloproteinase (TIMP)-2, and TIMP-3 were measured using Western blot analysis. Matrix metalloproteinase (MMP)-2 and MMP-9 activity was determined with gelatin zymography. Wound healing assay was used to determine cell migration. RESULTS: Punicalagin inhibited the cell viability of A2780 cells in a dose- and time-dependent manner, and the cell cycle of A2780 cells was arrested in G1/S phase transition. The treatment also induced apoptosis as shown by the up-regulation of Bax and down-regulation of Bcl-2. On the other hand, punicalagin treatment increased the expressions of TIMP-2 and TIMP-3, decreased the activities of MMP-2 and MMP-9, and inhibited cell migration. In addition, the ß-catenin pathway was suppressed as shown by the down-regulations of ß-catenin and its downstream factors including cyclin D1 and survivin. CONCLUSIONS: Punicalagin may have cancer-chemopreventive as well as cancer-chemotherapeutic effects against human ovarian cancer in humans through the inhibition of ß-catenin signaling pathway.

Characterization of Ca(2+)/H(+) exchange in the plasma membrane of Saccharomyces cerevisiae.

The characteristics of the Ca(2+)/H(+) exchange were directly investigated in functionally inverted (inside-out) plasma membrane vesicles isolated from yeast using an aqueous two-phase partitioning method. Results showed that following the generation of an inside-acid pH gradient (fluorescence quenching), addition of Ca(2+) caused movement of H(+) out of the vesicles (fluorescence recovery). The Ca(2+)/H(+) exchange displayed saturation kinetics with respect to extravesicular Ca(2+) and ATP concentrations in the plasma membrane, and showed specificity for Ca(2+). The protonophore FCCP (carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone), abolished the fluorescence quenching and consequently inhibited Ca(2+)/H(+) exchange in plasma membrane vesicles. Vanadate, which is known to inhibit the plasma membrane H(+)-ATPase, significantly decreased the Ca(2+)-dependent transport of H(+) out of vesicles. When the electrical potential across the plasma membrane was dissipated with valinomycin and potassium, the rate of Ca(2+)/H(+) exchange increased compared to that of the control without valinomycin, indicating that the stoichiometric ratio for this exchange is greater than 2H(+):Ca(2+). These data suggest that Ca(2+) is transported out of yeast cells through a Ca(2+)/H(+) exchange system that is driven by the proton-motive force generated by the plasma membrane H(+)-ATPase.

[Investigation of social support for inpatients with occupational diseases].

OBJECTIVE: To investigate and analyze the social support for inpatients with occupational diseases and to provide reference and basis for relevant medical and nursing interventions. METHODS: The social support rating scale (SSRS) was used to investigate the social support for 95 inpatients with occupational diseases. RESULTS: The total SSRS score of these patients was significantly lower than the national norm (32.5±9.31 vs 34.56±3.73, P < 0.05). The social support was mainly from the family, but medical staff and spiritual support were the main source and type of social support that are expected. CONCLUSION: Patients with occupational diseases have gained little social support, in both economic and spiritual aspects. In clinical practice, the patient's demand for knowledge of diseases and spiritual needs should be satisfied, and appropriate social support should be provided.

Angiopoietin-like protein 3: a novel potential biomarker for nephrotic syndrome in children.

Background: Angiopoietin-like 3 (ANGPTL3) is a secretory glycoprotein. It has been demonstrated that ANGPTL3 level was upregulated in minimal change nephrotic syndrome (MCNS) kidney tissues. Subsequently, our group found that ANGPTL3 level was closely correlated with nephropathy in vivo and in vitro. Hence, whether ANGPTL3 level could be correlated with the proteinuria level, and assessment of disease severity of nephrotic syndrome (NS) remained to be investigated. This study aimed to analyzed the correlation between the levels of ANGPTL3 in serum and urine of patients with nephrotic syndrome and proteinuria, and assessed the severity of the patients' disease. In future clinical translation, the level of ANGPTL3 in serum, urine will be used as a biomarker to better predict the development of nephrotic syndrome. Methods: A total of 200 NS patients and 80 healthy controls (age, 1-18 years) were admitted to our institution between 2021 and 2022. The etiology of NS included primary nephrotic syndrome (PNS, n = 144) and NS with other causes (n = 56). A total of 280 serum samples and 244 urinary samples were collected to determine ANGPTL3 level using enzyme-linked immunosorbent assay (ELISA). Results: Serum ANGPTL3 and urinary ANGPTL3/Cre were remarkably elevated in NS patients compared with those in healthy controls. Furthermore, serum ANGPTL3 and urinary ANGPTL3/Cre were significantly correlated with proteinuria level. Additionally, multivariate linear regression analysis demonstrated that serum ALB was independently correlated with serum ANGPTL3 and PRO/CR was independently correlated with urinary ANGPTL3/Cre in NS patients. Conclusion: Serum ANGPTL3 and urinary ANGPTL3/Cre showed a promising performance in the diagnosis of NS, and served as novel potential noninvasive biomarkers to assess disease severity of NS. Further exploration of the role of ANGPTL3 level may shed a new light on the treatment of NS.

Reproduction, cultural symbolism, and online relationship: Constructing city spatial imagery on TikTok.

The city on social media has become a hot topic in the study of city communication and city image. From the perspective of spatial theory and the communication characteristics of social media, this paper divides the spatial imagery of TikTok into three spaces: material space-cognitive attention, mental space-mental association, and relational space-emotional involvement. Based on the content analysis of 40 videos, we analyze the process of social media using cognition, association, and emotion as the starting points to increase the material space, expand the mental space, and expand the relational space. We find that spatial imagery can be co-constructed from the material space, mental space, and relational space. Lastly, the model is changed, and the value of using spatial theory to understand how city images are made is talked about.

Design and study of nitric oxide portable producing device using continuous discharging arc plasma reaction keeping low energy efficiency for viral pneumonia emergency therapy.

This study investigated the efficiency of a portable nitric oxide (NO) inhalation device through optimizing its design and structure. The portable rescue device could be used in clinical applications in outbreaks of viral pneumonia such as SARS. To reduce energy consumption for battery-powered portable usage, NO micro-channel plasma reactions induced by a continuous discharge arc were employed. A single-use airway tube could be combined with an intubation tube in clinical applications. In the experiment, a switching transistor controlled high frequency DC (12.5 kHz) was used to create a continuous discharge arc between two stainless steel electrodes (1-mm separation) after high-voltage breakthrough. A rotate instrument was employed to change the direction angle between the airflow and discharge arc, tube filled with Calcium hydroxide connected with gas outlet for reducing NO2, gas flow rate and input voltage were evaluated separately with concentration of NO and NO2/NO ratio. Results showed that a 2 L/min air flow direction from the cathode to the anode of electrodes (direction angle was zero) under 4 V input voltages produced 32.5±3.8 ppm NO, and the NO2/NO ratio reduced to less than 10%, stable output of nitric oxide might be convenient and effective for NO inhalation therapy. Modularization of the design produced a portable NO inhalation device that has potential for use in clinical applications as it is low cost, easy to disinfect, consumes low levels of energy and is ready to use.

Clinical utility in infants with suspected monogenic conditions through next-generation sequencing.

BACKGROUND: Rare diseases are complex disorders with huge variability in clinical manifestations. Decreasing cost of next-generation sequencing (NGS) tests in recent years made it affordable. We witnessed the diagnostic yield and clinical use of different NGS strategies on a myriad of monogenic disorders in a pediatric setting. METHODS: Next-generation sequencing tests are performed for 98 unrelated Chinese patients within their first year of life, who were admitted to Xin Hua Hospital, affiliated with Shanghai Jiao Tong University School of Medicine, during a 2-year period. RESULTS: Clinical indications for NGS tests included a range of medical concerns. The mean age was 4.4 ± 4.2 months of age for infants undergoing targeting specific (known) disease-causing genes (TRS) analysis, and 4.4 ± 4.3 months of age for whole-exome sequencing (WES) (p > 0.05). A molecular diagnosis is done in 72 infants (73.47%), which finds a relatively high yield with phenotypes of metabolism/homeostasis abnormality (HP: 0001939) (odds ratio, 1.83; 95% CI, 0.56-6.04; p = 0.32) and a significantly low yield with atypical symptoms (without a definite HPO term) (odds ratio, 0.08; 95% CI, 0.01-0.73; p = 0.03). TRS analysis provides molecular yields higher than WES (p = 0.01). Ninety-eight different mutations are discovered in 72 patients. Twenty-seven of them have not been reported previously. Nearly half (43.06%, 31/72) of the patients are found to carry 11 common disorders, mostly being inborn errors of metabolism (IEM) and neurogenetic disorders and all of them are observed through TRS analysis. Eight positive cases are identified through WES, and all of them are sporadic, of highly variable phenotypes and severity. There are 26 patients with negative findings in this study. CONCLUSION: This study provides evidence that NGS can yield high success rates in a tertiary pediatric setting, but suggests that the scope of known Mendelian conditions may be considerably broader than currently recognized.

Effect of integrin ß1 in the treatment of stress urinary incontinence by electrical stimulation.

The aim of the present study was to investigate the protective effect of integrin ß1 in the treatment of stress urinary incontinence (SUI) by electrical stimulation, and the underlying mechanisms by which electrical stimulation regulates the collagen metabolism of female vaginal wall fibroblasts (FVWFs). FVWFs obtained from the vaginal wall tissue of patients with (Ingelman­Sundberg scale; grade II, n=8; grade III, n=10) or without (n=8) SUI during gynecological operations were isolated by enzymatic digestion and subsequently identified by immunocytochemistry. Following this, cultured FVWFs were treated with an inhibitor of integrin ß1, recombinant human integrin ß1 and electrical stimulation (100 mv/mm, 2 h, 20 Hz), followed by total mRNA and protein extraction. mRNA and protein expression levels of integrin ß1, transforming growth factor (TGF)­ß1 and collagen (COL) I and III in FVWFs were quantified by reverse transcription­quantitative PCR (RT­qPCR) and western blot analysis respectively. Integrin ß1, TGF­ß1 and COL I and III expression levels were decreased in patients with SUI compared with healthy controls, and the grade III group had lower levels than the grade II group. Following electrical stimulation treatment, the expression levels of TGF­ß1, COL I and III were enhanced in the grade II group, but not in the grade III group. Nevertheless, the inhibitor of integrin ß1 reduced the protective effect of electrical stimulation in the grade II group. In addition, electrical stimulation combined with recombinant human integrin ß1 could also protect cells from SUI in the grade III group. The present study provides evidence for the increased degradation of the extracellular matrix and integrin ß1 in the vaginal wall tissues of patients with SUI, and the protective effect of electrical stimulation against SUI via integrin ß1. These results provide a novel mechanism for the treatment of SUI using electrical stimulation.

Effect of peroxisome proliferator-activated receptors-gamma and co-activator-1alpha genetic polymorphisms on plasma adiponectin levels and susceptibility of non-alcoholic fatty liver disease in Chinese people.

BACKGROUND/AIMS: Peroxisome proliferator-activated receptors-gamma (PPAR-gamma) and its co-activator-1alpha (PGC-1alpha) are involved in the regulation of lipid and glucose metabolisms. This study aimed to investigate the genetic polymorphisms of PPAR-gamma and PGC-1alpha in Chinese people and their influence on plasma adiponectin levels and non-alcoholic fatty liver disease (NAFLD) susceptibility. METHODS: Ninety-six patients with NAFLD and 96 healthy controls were included. The single nucleotide polymorphisms (SNPs) of C161T PPAR-gammaand Gly482Ser PGC-1alpha genes were analysed by polymerase chain reaction and restriction fragment length polymorphism. RESULT: The CC, CT and TT genotypic distributions of the NAFLD group were significantly different from those of controls (55.2, 39.6, 5.2 vs. 74.0, 25.0, 1.0%; P=0.015). The allelic frequencies of C and T were also different between the two groups (P=0.004). As for the PGC-1alpha gene, there was no difference of the genotypic and allelic frequencies between the two groups (P>0.05). In NAFLD patients, the plasma adiponectin concentrations were lower in the PPAR-gamma CT/TT genotypes compared with those in the CC genotype group (3.0+/-0.6 vs. 4.3+/-0.9, P=0.02). Multivariate logistic regression analysis showed that CT/TT genotypes of PPAR-gamma, TG, waist hip ratio, hypoadiponectinaemia and homoeostasis model assessment (HOMA)-IR were the risk factors for NAFLD. CONCLUSION: SNPs in the PPAR-gamma, but not PGC-1alpha, gene are associated with NAFLD susceptibility possibly through the adiponectin pathway.

Role of transforming growth factor ß­1 in the pathogenesis of pelvic organ prolapse: A potential therapeutic target.

The present study aimed to reveal the metabolic alterations of the extracellular matrix (ECM) in uterosacral ligament (USL) with pelvic organ prolapse (POP) and to explore the role of transforming growth factor­ß1 (TGF­ß1) in pathogenesis of POP. For this purpse, 60 participants who underwent hysterectomy for benign indications were enrolled, 30 of which had symptomatic POP (grade II, III or IV) and composed the POP group, and the other 30 had asymptomatic POP (grade I or less) and served as the controls. Collagen fibers, elastin,matrix metalloproteinase (MMP)­2/9, tissue inhibitor of matrix metalloproteinases (TIMP)­2 and TGF­ß1 were examined by Masson's trichrome staining, immunohistochemistry and RT-qPCR using USL biopsies. In vitro, human USL fibroblasts (hUSLFs) were primary cultured, pre-treated with recombinant TGF­ß1 (0, 5, or 10 ng/ml) and then subjected to cyclic mechanical stretching (CMS; 0 or 5,333 µÎµ strain). Changes in the expression levels of collagen type I/III, elastin, TIMP­2, MMP­2/9 and Smad were detected. Our results revealed that at the tissue level, the expression of collagen fibers, elastin, TIMP­2 and TGF­ß1 was significantly reduced in the POP group, while the activities of MMP­2/9 were significantly upregulated, compared with the control group. Statistical analysis indicated that the mRNA expression of TGF­ß1 inversely correlated with the severity of POP partially. Our in vitro experimental data demonstrated that a CMS of 5333 µÎµ strain promoted the degradation of ECM proteins, inhibited the synthesis of TIMP­2, and upregulated the proteolytic activities of MMP­2/9. Pre-treatment with TGF­ß1 attenuated the loss of ECM by stimulating the synthesis of TIMP­2 and inhibiting the activities of MMP­2/9 through the TGF­ß1/Smad3 signaling pathway. On the whole, our data indicate that the reduced anabolism and increased catabolism of ECM proteins in USL are the pathological characteristics of POP. TGF­ß1 not only has a specific value in predicting the severity of POP, but should also be considered as a novel therapeutic target for POP.

Optimum design of electrode structure and parameters in electrical impedance tomography.

In electrical impedance tomography (EIT) the electrode structure and parameters significantly influence measurement sensitivity and image quality, so how to optimize the electrode structure and parameters is one of the key problems in research today. This paper presents a method to optimize the EIT electrode structure and parameters based on coercive equipotential node models. The coercive equipotential mode of the compound electrode has been established based on that of the line electrode. A simulation study for the line electrode and the compound electrode of EIT has been made on a simulation software platform. The influences of different electrode structures and parameters on measurement sensitivity and the image reconstruction quality are studied. For line electrode simulation studies, two important conclusions are drawn. First, a narrower electrode is helpful in improving the imaging quality. Second, although it is known that a wider electrode is beneficial in decreasing the contact impedance, using a too wide electrode causes the measurement sensitivity to decrease. Furthermore the electrode width that leads to the best measurement sensitivity is different for different measurement depths. The compound electrode has four parameters: the excitation electrode width, the measurement electrode width, the space between the excitation electrode and the measurement electrode, and the distance between two adjacent compound electrodes. These parameters have mutual restrictions and complex influences on each other. It is unwise to optimize the design of a compound electrode by only using the overlay rate of electrodes. A simulation study of EIT electrode structure and parameter influences can be carried out according to this paper to determine the optimum design of the electrode structure and its parameters.

Role of mechanical strain-activated PI3K/Akt signaling pathway in pelvic organ prolapse.

Mechanical loading on pelvic supports contributes to pelvic organ prolapse (POP). However, the underlying mechanisms remain to be elucidated. Our previous study identified that mechanical strain induced oxidative stress (OS) and promoted apoptosis and senescence in pelvic support fibroblasts. The aim of the present study is to investigate the molecular signaling pathway linking mechanical force with POP. Using a four­point bending device, human uterosacral ligament fibroblasts (hUSLF) were exposed to mechanical tensile strain at a frequency of 0.3 Hz and intensity of 5333 µÎµ, in the presence or absence of LY294002. The applied mechanical strain on hUSLF resulted in apoptosis and senescence, and decreased expression of procollagen type I α1. Mechanical strain activated phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt signaling and resulted in downregulated expression of glutathione peroxidase 1 and Mn­superoxide dismutase, and accumulation of intracellular reactive oxygen species. These effects were blocked by administration of LY294002. Furthermore, it was demonstrated that PI3K/Akt was activated in the uterosacral ligaments of POP patients, and that OS was increased and collagen type I production reduced. The results from the present study suggest that mechanical strain promotes apoptosis and senescence, and reduces collagen type I production via activation of PI3K/Akt-mediated OS signaling pathway in hUSLF. This process may be involved in the pathogenesis of POP as it results in relaxation and dysfunction of pelvic supports.

Collagen metabolic disorder induced by oxidative stress in human uterosacral ligament­derived fibroblasts: A possible pathophysiological mechanism in pelvic organ prolapse.

Pelvic organ prolapse (POP) is a global health problem, for which the pathophysiological mechanism remains to be fully elucidated. The loss of extracellular matrix protein has been considered to be the most important molecular basis facilitating the development of POP. Oxidative stress (OS) is a well­recognized mechanism involved in fiber metabolic disorders. The present study aimed to clarify whether OS exists in the uterosacral ligament (USL) with POP, and to investigate the precise role of OS in collagen metabolism in human USL fibroblasts (hUSLFs). In the present study, 8­hydroxyguanosine (8­OHdG) and 4 hydroxynonenal (4­HNE), as oxidative biomarkers, were examined by immunohistochemistry to evaluate oxidative injury in USL sections in POP (n=20) and non­POP (n=20) groups. The primary cultured hUSLFs were treated with exogenous H2O2 to establish an original OS cell model, in which the expression levels of collagen, type 1, α1 (COL1A1), matrix metalloproteinase (MMP)­2, tissue inhibitor of metalloproteinase (TIMP)­2 and transforming growth factor (TGF)­ß1 were evaluated by western blot and reverse transcription­quantitative polymerase chain reaction analyses. The results showed that the expression levels of 8­OHdG and 4­HNE in the POP group were significantly higher, compared with those in the control group. Collagen metabolism was regulated by H2O2 exposure in a concentration­dependent manner, in which lower concentrations of H2O2 (0.1­0.2 mM) stimulated the anabolism of COL1A1, whereas a higher concentration (0.4 mM) promoted catabolism. The expression levels of MMP­2, TIMP­2 and TGF­ß1 exhibited corresponding changes with the OS levels. These results suggested that OS may be involved in the pathophysiology of POP by contributing to collagen metabolic disorder in a severity­dependent manner in hUSLFs, possibly through the regulation of MMPs, TIMPs and TGF­ß1 indirectly.

Functional characterization of a wheat NHX antiporter gene TaNHX2 that encodes a K(+)/H(+) exchanger.

The subcellular localization of a wheat NHX antiporter, TaNHX2, was studied in Arabidopsis protoplasts, and its function was evaluated using Saccharomyces cerevisiae as a heterologous expression system. Fluorescence patterns of TaNHX2-GFP fusion protein in Arabidopsis cells indicated that TaNHX2 localized at endomembranes. TaNHX2 has significant sequence homology to NHX sodium exchangers from Arabidopsis, is abundant in roots and leaves and is induced by salt or dehydration treatments. Western blot analysis showed that TaNHX2 could be expressed in transgenic yeast cells. Expressed TaNHX2 protein suppressed the salt sensitivity of a yeast mutant strain by increasing its K(+) content when exposed to salt stress. TaNHX2 also increased the tolerance of the strain to potassium stress. However, the expression of TaNHX2 did not affect the sodium concentration in transgenic cells. Western blot analysis for tonoplast proteins indicated that the TaNHX2 protein localized at the tonoplast of transgenic yeast cells. The tonoplast vesicles from transgenic yeast cells displayed enhanced K(+)/H(+) exchange activity but very little Na(+/)H(+) exchange compared with controls transformed with the empty vector; Na(+)/H(+) exchange was not detected with concentrations of less than 37.5 mM Na(+) in the reaction medium. Our data suggest that TaNHX2 is a endomembrane-bound protein and may primarily function as a K(+)/H(+) antiporter, which is involved in cellular pH regulation and potassium nutrition under normal conditions. Under saline conditions, the protein mediates resistance to salt stress through the intracellular compartmentalization of potassium to regulate cellular pH and K(+) homeostasis.

[The mechanism for up-regulation of HLA-I expression on HepG2 cells by HBV].

AIM: To explore the mechanism of up-regulation of HLA-I expression on HepG2 cells by wild type (WT) and nucleocapsid mutants(L97 and V60) of hepatitis B virus (HBV). METHODS: The HBV-stable expression vectors EBO-WT, EBO-L97 and EBO-V60 were transfected into HepG2 cells via the liposome mediation, respectively. The cells were assayed by semi-quantitative RT-PCR for HLA-A gene and antigen presentation-related genes LMP2, TAP1, and tapasin mRNA expression. Western blot was applied for analysis of HLA-I protein expression in the cells. RESULTS: HepG2 cells transfected by the 3 HBV expression vectors expressed HLA-A and TAP1, while there was no expression of HLA-A and only marginal expression of TAP1 in HepG2 cells transfected by control vector EBO. The expression level of HLA-A in the transfected cells decreased successively in the order of EBO-L97, EBO-WT and EBO-V60. There was no significant difference in the expression level of TAP1 between HepG2 cells transfected by the 3 HBV expression vectors. No detectable expression of LMP2 and tapasin was observed for all transfected cells. CONCLUSION: HBV can induce the expression of HLA-I molecule and TAP1 in HepG2 cells.