A circulating microRNA panel as a novel dynamic monitor for oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) has high recurrence and mortality rates despite advances in diagnosis and treatment. Therefore, it is necessary to identify new biomarkers for early detection, efficient monitoring, and prognosis prediction. Since microRNA (miRNA) is stable and detectable in serum, it has been reported to inform the diagnosis and monitor disease progression through liquid biopsy. In this study, a circulating specific miRNA panel in OSCC patients was developed, and its usefulness as a dynamic monitor was validated. Small RNAs were extracted from the serum of OSCC patients (n = 4) and normal controls (n = 6) and profiled using next-generation sequencing. NGS identified 42 differentially expressed miRNAs (DEmiRNAs) in serum between patients with OSCC and healthy controls, with threefold differences (p < 0.05). Combining the 42 DEmiRNAs and The Cancer Genome Atlas (TCGA) databases OSCC cohort, 9 overlapping DEmiRNAs were screened out. Finally, 4 significantly up-regulated miRNAs (miR-92a-3p, miR-92b-3p, miR-320c and miR-629-5p) were identified from OSCC patients via validation in the Chungnam National University Hospital cohort. Application of the specific miRNA panel for distinguishing OSCC patients from healthy controls produced specificity and sensitivity of 97.8 and 74%, respectively. In addition, the serum levels of these 4 miRNAs significantly decreased after complete surgical resection and increased after recurrence. We suggest that circulating 4-miRNA panel might be promising non-invasive predictors for diagnosing and monitoring the progression of patients with OSCC.

Claudin-1 mediates progression by regulating EMT through AMPK/TGF-ß signaling in head and neck squamous cell carcinoma.

Claudin-1 (CLDN1), a major component of tight junction complexes in the epithelium, maintains cellular polarity, and plays a critical role in cell-to-cell communication as well as epithelial cell homeostasis. Although the role of CLDN1 has been widely studied in cancer, its role in the progression and the exact regulatory mechanisms, remain controversial. Using next-generation sequencing, we first analyzed the expression profiles of tumor/non-tumor paired tissue in patients with head and neck squamous cell carcinoma (HNSC) from public and local cohorts and found out that CLDN1 is upregulated in tumors compared to normal tissues. Next, its correlation with lymph node metastasis and poor prognosis was validated in the retrospective cohort, which collectively suggests CLDN1 as an oncogene in HNSC. As expected, the knockdown of CLDN1 inhibited invasive phenotypes by downregulating epithelial-to-mesenchymal transition (EMT) in vitro. To ascertain the regulatory mechanism of CLDN1 in HNSC analysis of GO term enrichment, KEGG pathways, and curated gene sets were used. As a result, CLDN1 was negatively associated with AMP-activated protein kinase (AMPK) and positively associated with transforming growth factor-ß (TGF-ß) signaling. In vitro mechanistic assay showed that CLDN1 inhibited AMPK phosphorylation by regulating AMPK upstream phosphatases, which led to inhibition of Smad2 activity. Intriguingly, the invasive phenotype of cancer cells increased by CLDN1 overexpression was rescued by AMPK activation, indicating a role of the CLDN1/AMPK/TGF-ß/EMT cascade in HNSC. Consistently in vivo, CLDN1 suppression significantly inhibited the tumor growth, with elevated AMPK expression, suggesting the novel observation of oncogenic CLDN1-AMPK signaling in HNSC.

Key mutations on spike protein altering ACE2 receptor utilization and potentially expanding host range of emerging SARS-CoV-2 variants

Increasing evidence supports inter-species transmission of SARS-CoV-2 variants from human to domestic or wild animals during the ongoing COVID-19 pandemic, which is posing great challenges to epidemic control. Clarifying the host range of emerging SARS-CoV-2 variants will provide instructive information for the containment of viral spillover. The spike protein (S) of SARS-CoV-2 is the key determinant of receptor utilization, and therefore amino acid mutations on S will probably alter viral host range. Here, in order to evaluate the impact of S mutations, we constructed 20 Hela cell lines stably expressing ACE2 orthologs from different animals, and prepared 27 pseudotyped SARS-CoV-2 carrying different spike mutants, among which 20 bear single mutation and the other 7 were cloned from emerging SARS-CoV-2 variants, including D614G, Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.135), Lambda (B.1.429) and Mu (B.1.525). Using pseudoviral reporter assay, we identified that the substitutions of T478I and N501Y enabled the pseudovirus to utilize chicken ACE2, indicating potential infectivity to avian species. Furthermore, the S mutants of real SARS-CoV-2 variants comprising N501Y showed significantly acquired abilities to infect cells expressing mouse ACE2, indicating a critical role of N501Y in expanding SARS-CoV-2 host range. In addition, A262S and T478I significantly enhanced the utilization of various mammals ACE2. In summary, our results indicated that T478I and N501Y substitutions were two S mutations important for receptor adaption of SARS-CoV-2, potentially contributing to spillover of the virus to many other animal hosts. Therefore, more attention should be paid to SARS-CoV-2 variants with these two mutations.

The effect of Curcumin on multi-level immune checkpoint blockade and T cell dysfunction in head and neck cancer.

BACKGROUND: Despite recent advances in understanding the complex immunologic dysfunction in the tumor microenvironment (TME), fewer than 20% of patients with head and neck squamous cell carcinoma (HNSCC) respond to immune checkpoint blockade (ICB). Thus, it is important to understand how inhibitory IC receptors maintain the suppressed dysfunctional TME, and to develop more effective combination immunotherapy. This study evaluated the immune-modulating effects of Curcumin, which has well-established anti-cancer and chemopreventive properties, and its long-term safety as a phytochemical drug. METHODS: We carried out the western blot and small interfering RNA (siRNA) transfection assay to evaluate the effects of Curcumin on IC ligands and IC ligands function in HNSCC. Through T-cell cytotoxicity assay and measurements of cytokine secretion, we assessed the effects of combination of Curcumin with programmed death-ligand 1 (PD-L1) Ab on cancer cell killing. Flow cytometry were used to analyze the effects of Curcumin on the expression of programmed cell death protein 1 (PD-1) and T-cell immunoglobulin and mucin-domain3 (TIM-3) on CD4, CD8 and Treg. Immunofluorescence, immunohistochemistry and western blot were used to detecte the cytokine (IFN-γ, Granzyme B), IC receptors (PD-1 and TIM-3) and its ligands (PD-L1, PD-L2, Galectin-9) in xenograft mouse model and 4-nitroquinoline-1-oxide (4-NQO) oral cancer model. RESULTS: We found that Curcumin decreased the expression of IC ligands such as PD-L1, PD-L2, and Galectin-9 in HNSCC, leading to regulation of epithelial-to-mesenchymal transition-associated tumor invasion. Curcumin also effectively restored the ability of CD8+ cytotoxic T cells to lyse cancer cells. To evaluate the effect of Curcumin on the TME further, the 4-NQO oral cancer model was used. Curcumin increased T-cell proliferation, tumor-infiltrating lymphocytes (TILs), and effector cytokines, and decreased the expression of PD-1, TIM-3, suppressive IC receptors and their ligands (PD-L1, PD-L2, and Galectin-9) in the TME, implying reinvigoration of the exhausted CD8+ T cells. In addition, Curcumin inhibited expression of CD4+CD25+FoxP3+ Treg cells as well as PD-1 and TIM-3. CONCLUSIONS: These results show that Curcumin reinvigorates defective T cells via multiple (PD-1 and TIM-3) and multi-level (IC receptors and its ligands) IC axis suppression, thus providing a rationale to combine Curcumin with conventional targeted therapy or ICB as a multi-faceted approach for treating patients with HNSCC.

Head and Neck Cancer Cell Death due to Mitochondrial Damage Induced by Reactive Oxygen Species from Nonthermal Plasma-Activated Media: Based on Transcriptomic Analysis.

Mitochondrial targeted therapy is a next-generation therapeutic approach for cancer that is refractory to conventional treatments. Mitochondrial damage caused by the excessive accumulation of reactive oxygen species (ROS) is a principle of mitochondrial targeted therapy. ROS in nonthermal plasma-activated media (NTPAM) are known to mediate anticancer effects in various cancers including head and neck cancer (HNC). However, the signaling mechanism of HNC cell death via NTPAM-induced ROS has not been fully elucidated. This study evaluated the anticancer effects of NTPAM in HNC and investigated the mechanism using transcriptomic analysis. The viability of HNC cells decreased after NTPAM treatment due to enhanced apoptosis. A human fibroblast cell line and three HNC cell lines were profiled by RNA sequencing. In total, 1 610 differentially expressed genes were identified. Pathway analysis showed that activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP) were upstream regulators. Mitochondrial damage was induced by NTPAM, which was associated with enhancements of mitochondrial ROS (mtROS) and ATF4/CHOP regulation. These results suggest that NTPAM induces HNC cell death through the upregulation of ATF4/CHOP activity by damaging mitochondria via excessive mtROS accumulation, similar to mitochondrial targeted therapy.

Water resources allocation model based on ecological priority in the arid region.

Based on the characteristics of arid regions in the central and southern Ningxia，firstly we constructed the theoretical framework about rational allocation of water resources based on ecological priority, which supplemented and improved the theoretical system on rational allocation of water resources for sustainable development. On the basis of theoretical research, depending on its characteristic of water resources：pumping water from Yellow River in the north, piloting water from Jing River in the south and regional water resources, which formed a mixing water supply pattern of "pumping, piloting, storage". The theory of water resource allocation system was perfected. Then the mathematical model for the rational allocation of water resources was established on the basis of biggest benefit combined with economy, society and ecology. The model coincided the ecological water and domestic water benefit into the overall benefits, and provides them larger benefit coefficient and the priority order coefficient to realize the reasonable allocation of water resources on ecological priority mode. Finally, the results of allocation are analysed in detail, which provides constructive suggestions and guiding direction for the study of the rational allocation of regional water resources.

EGR1/GADD45α Activation by ROS of Non-Thermal Plasma Mediates Cell Death in Thyroid Carcinoma.

(1) Background: Nonthermal plasma (NTP) induces cell death in various types of cancer cells, providing a promising alternative treatment strategy. Although recent studies have identified new mechanisms of NTP in several cancers, the molecular mechanisms underlying its therapeutic effect on thyroid cancer (THCA) have not been elucidated. (2) Methods: To investigate the mechanism of NTP-induced cell death, THCA cell lines were treated with NTP-activated medium -(NTPAM), and gene expression profiles were evaluated using RNA sequencing. (3) Results: NTPAM upregulated the gene expression of early growth response 1 (EGR1). NTPAM-induced THCA cell death was enhanced by EGR1 overexpression, whereas EGR1 small interfering RNA had the opposite effect. NTPAM-derived reactive oxygen species (ROS) affected EGR1 expression and apoptotic cell death in THCA. NTPAM also induced the gene expression of growth arrest and regulation of DNA damage-inducible 45α (GADD45A) gene, and EGR1 regulated GADD45A through direct binding to its promoter. In xenograft in vivo tumor models, NTPAM inhibited tumor progression of THCA by increasing EGR1 levels. (4) Conclusions: Our findings suggest that NTPAM induces apoptotic cell death in THCA through a novel mechanism by which NTPAM-induced ROS activates EGR1/GADD45α signaling. Furthermore, our data provide evidence that the regulation of the EGR1/GADD45α axis can be a novel strategy for the treatment of THCA.

Brn3a/Pou4f1 Functions as a Tumor Suppressor by Targeting c-MET/STAT3 Signaling in Thyroid Cancer.

BACKGROUND: Brn3a/Pou4f1 is a class IV POU domain-containing transcription factor and has been found to be expressed in a variety of cancers. However, the mechanism and action of Brn3a in thyroid cancer has not been investigated. PURPOSE: To investigate the role of Brn3a in thyroid cancer progression and its clinical implication. METHODS: We examined Brn3a expression status in patients with thyroid cancer and analyzed relationships between Brn3a expression and clinicopathological findings using The Cancer Genome Atlas (TCGA) database. For functional in vitro analysis, proliferation, migration, invasion assay, and Western blotting were performed after overexpression or suppression of Brn3a. RESULTS: The promoter hypermethylation of Brn3a was found in patients with aggressive thyroid cancer and Brn3a was downregulated in tissues of patients with thyroid cancer. In TCGA database, the low-Brn3a-expression group revealed a more aggressive phenotype, including T stage and extrathyroid extension when compared with the high-Brn3a-expression group. Overexpression of Brn3a suppressed cell migration and invasion via regulation of epithelial-mesenchymal transition (EMT)-associated proteins in thyroid cancer cell lines. Brn3a overexpression also downregulated signal transducer and activator of transcription 3 (STAT3) signaling through suppression of tyrosine-protein kinase Met (c-MET). In contrast, knockdown of Brn3a by small interfering ribonucleic acid (siRNA) significantly increased cell migration and invasion through upregulation of c-MET/STAT3. These results imply that Brn3a suppresses tumor metastasis via c-MET/STAT3 inhibition and EMT suppression in thyroid cancer. CONCLUSIONS: Our findings show that Brn3a is a potential tumor suppressor that leads to reduced cancer cell migration and invasion in thyroid cancer. Elucidation of the Brn3a-regulated cancer pathways may therefore provide novel therapeutic strategies to control thyroid cancer metastasis.

Receptor utilization of angiotensin converting enzyme 2 (ACE2) indicates a narrower host range of SARS-CoV-2 than that of SARS-CoV

Coronavirus pandemics have become a huge threat to the public health worldwide in the recent decades. Typically, SARS-CoV caused SARS pandemic in 2003 and SARS-CoV-2 caused the COVID-19 pandemic recently. Both viruses have been reported to originate from bats. Thus, direct or indirect interspecies transmission from bats to humans is required for the viruses to cause pandemics. Receptor utilization is a key factor determining the host range of viruses which is critical to the interspecies transmission. Angiotensin converting enzyme 2 (ACE2) is the receptor of both SARS-CoV and SARS-CoV-2, but only ACE2s of certain animals can be utilized by the viruses. Here, we employed pseudovirus cell-entry assay to evaluate the receptor-utilizing capability of ACE2s of 20 animals by the two viruses and found that SARS-CoV-2 utilized less ACE2s than SARS-CoV, indicating a narrower host range of SARS-CoV-2. Meanwhile, pangolin CoV, another SARS-related coronavirus highly homologous to SARS-CoV-2 in its genome, yet showed similar ACE2 utilization profile with SARS-CoV rather than SARS-CoV-2. To clarify the mechanism underlying the receptor utilization, we compared the amino acid sequences of the 20 ACE2s and found 5 amino acid residues potentially critical for ACE2 utilization, including the N-terminal 20th and 42nd amino acids that may determine the different receptor utilization of SARS-CoV, SARS-CoV-2 and pangolin CoV. Our studies promote the understanding of receptor utilization of pandemic coronaviruses, potentially contributing to the virus tracing, intermediate host screening and epidemic prevention for pathogenic coronaviruses.

Effectiveness of percutaneous neuromuscular electrical stimulation for neck pain relief in patients with cervical spondylosis.

BACKGROUND: This study aimed to evaluate the effectiveness and safety of percutaneous neuromuscular electrical stimulation (PNMES) for treating neck pain in patients with cervical spondylosis (CS). METHODS: One hundred and twenty four patients with neck pain of CS were included, and then they were randomly divided into a PNMES group and a control group in a ratio of 1:1. All patients received PNMES or sham PNMES 30 minutes daily, 3 times weekly for 12 weeks. The primary outcome was assessed by the visual analog scale (VAS). The secondary outcomes were evaluated by the cervical range of motion (ROM), neck disability index (NDI) score, as well as the adverse events (AEs). All outcome measurements were measured at the end of 12-week treatment, and 4-week follow-up after treatment. RESULTS: At the end of the 12-week treatment, and 4-week follow-up, the patients receiving PNMES exhibited more decrease in the mean VAS (P < .01), and NDI (P < .01) respectively, compared with the patients receiving sham PNMES. Additionally, the increase in the mean ROM was also significantly higher in the PNMES group than that in the sham PNMES group at the end of the 12-week treatment, and 4-week follow-up, respectively (P < .01). No AEs were found in either group. CONCLUSIONS: The results of this study demonstrated that PNMES is more effective than Sham PNMES for neck pain relief in patients with CS.

Overexpression of Populus trichocarpa CYP85A3 promotes growth and biomass production in transgenic trees.

Brassinosteroids (BRs) are essential hormones that play crucial roles in plant growth, reproduction and response to abiotic and biotic stress. In Arabidopsis, AtCYP85A2 works as a bifunctional cytochrome P450 monooxygenase to catalyse the conversion of castasterone to brassinolide, a final rate-limiting step in the BR-biosynthetic pathway. Here, we report the functional characterizations of PtCYP85A3, one of the three AtCYP85A2 homologous genes from Populus trichocarpa. PtCYP85A3 shares the highest similarity with AtCYP85A2 and can rescue the retarded-growth phenotype of the Arabidopsis cyp85a2-2 and tomato dx mutants. Constitutive expression of PtCYP85A3, driven by the cauliflower mosaic virus 35S promoter, increased the endogenous BR levels and significantly promoted the growth and biomass production in both transgenic tomato and poplar. Compared to the wild type, plant height, shoot fresh weight and fruit yield increased 50%, 56% and 43%, respectively, in transgenic tomato plants. Similarly, plant height and stem diameter increased 15% and 25%, respectively, in transgenic poplar plants. Further study revealed that overexpression of PtCYP85A3 enhanced xylem formation without affecting the composition of cellulose and lignin, as well as the cell wall thickness in transgenic poplar. Our finding suggests that PtCYP85A3 could be used as a potential candidate gene for engineering fast-growing trees with improved wood production.

Characterization of Arabidopsis Tubby-like proteins and redundant function of AtTLP3 and AtTLP9 in plant response to ABA and osmotic stress.

Tubby and Tubby-like proteins (TLPs) play essential roles in the development and function of mammal neuronal cells. In addition to the conserved carboxyl (C)-terminal Tubby domain, which is required for their plasma membrane (PM) tethering, plant TLPs also possess an amino (N)-terminal F-box domain to interact with specific Arabidopsis Skp1-like (ASK) proteins as functional SCF-type E3 ligases. Here, we report the molecular characterization of Arabidopsis TLPs (AtTLPs). ß-Glucuronidase staining showed overlapped but distinct expression patterns of AtTLPs in Arabidopsis. Yeast two-hybrid assays further revealed that AtTLP1, AtTLP3, AtTLP6, AtTLP7, AtTLP9, AtTLP10 and AtTLP11 all interacted with specific ASKs, but AtTLP2, AtTLP5 and AtTLP8 did not. Subcellular localization observations in both Arabidopsis protoplasts and tobacco pollen tubes indicated that all GFP-AtTLP fusion proteins, except GFP-AtTLP8 which lacks the conserved phosphatidylinositol 4,5-bisphosphate binding sites, were targeted to the PM. Detailed studies on AtTLP3 demonstrated that AtTLP3 is a PM-tethered PIP2 binding protein which functions redundantly with AtTLP9 in abscisic acid (ABA)- and osmotic stress-mediated seed germination. Our results suggest that AtTLPs possibly work in multiple physiological and developmental processes in Arabidopsis, and AtTLP3 is also involved in ABA signaling pathway like AtTLP9 during seed germination and early seedling growth.

Effects of subchronic benzoapyrene exposure on hippocampal cholinergic system in rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the effects of subchronic benzo[a]pyrene (B[a]P) exposure on the neurobehavior and hippocampal acetylcholine (Ach) level, acetylcholinesterase (AChE) activity, and mRNA and protein expression of nicotinic acetylcholine receptor α7 subtype (nAChR α7) in rats, and to investigate the neurotoxic mechanism of B[a]P.</p><p><b>METHODS</b>Sixty healthy male SD rats were randomly divided into blank control group, solvent control group, and B [a]P exposure groups. Each rat in the exposure groups was intraperitoneally injected with B[a]P at 1.0, 2.5, or 6.25 mg/kg once every other day for 90 days. The learning and memory ability of the rats was examined by Morris water maze test and step-down test; the hippocampal Ach level was measured by alkaline hydroxylamine method; the AChE activity was measured by DNTB method; the mRNA and protein expression levels of hippocampal nAChR α7 were measured by quantitative PCR and Western blot.</p><p><b>RESULTS</b>The 2.5 and 6.25 mg/kg B[a]P exposure groups showed significantly lower learning and memory abilities than the blank control group and solvent control group (P < 0.05); also, the two groups had significantly lower hippocampal Ach levels than the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group (P < 0.05). The 6.25 mg/kg B[a]P exposure group showed significantly lower hippocampal AChE activity than the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group (P < 0.05). There were no significant differences in the mRNA and protein expression levels of nAChR α7 among all groups (P > 0.05). The hippocampal Ach level was negatively correlated with the mean escape latency period and total distance travelled (r = -0.567, P < 0.01; r = -0.503, P < 0.01) but positively correlated with the time in platform quadrant (r = 0.800, P < 0.01).</p><p><b>CONCLUSION</b>Subchronic B[a]P exposure may impair the learning and memory ability in rats, which is related to the downregulation of hippocampal Ach level.</p>

The modification of apoptosis correlated gene, protein and protein activity in rat hippocampus induced by benzoa pyrene sub-chronic exposure / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the effects of subchronic exposure to benzo[a]pyrene (B[a]P) on the mRNA and protein expression levels of apoptosis-related genes (bax, bcl-2, caspase-3, caspase-6, and caspase-9) and the activities of Caspase-3, Caspase-6, and Caspase-9 in the hippocampal neurons of rats and to investigate the neurotoxic mechanism by which B[a]P induces the apoptosis of neurons.</p><p><b>METHODS</b>Fifty-two healthy SD rat were randomly divided into five groups according to preliminary neurobehavioral test results: blank control group, solvent control group, and 1.0, 2.5, and 6.25 mg/kg B[a]P exposure groups; the rats in exposure groups were intraperitoneally injected with B[a]P every other day for 90 days. The Morris water maze was used to test the learning and memory ability of rats; flow cytometry was used to measure the apoptosis ratio of hippocampal neurons; real-time quantitative PCR and Western blot were used to measure the mRNA and protein expression levels of apoptosis-related genes; spectrophotometry was used to measure the activities of their en-coded proteins.</p><p><b>RESULTS</b>Compared with the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group, the 2.5 and 6.25 mg/kg B[a]P exposure groups hada significantly longer mean escape latency period (P < 0.05) and a significantly increased number of times of platform crossing (P < 0.05), and the 6.25 mg/kg B[a]P exposure group had significantly lower length and percentage of time spent in the platform quadrant (P < 0.05). The early apoptosis ratio rose as the dose of B[a]P increased (P trend < 0.05); the early apoptosis ratios of 1.0, 2.5, and 6.25 mg/kg B[a]P exposure groups were significantly higher than those of blank control group and solvent control group (P < 0.05). Compared with the blank control group, solvent control group, and 1.0 and 2.5 mg/kg B[a]P exposure groups, the 6.25 mg/kg B[a]P exposure group had significantly increased Bax expression (P < 0.05) and significantly decreased Bcl-2 expression and Bcl-2/Bax ratio (P < 0.05). The 2.5 and 6.25 mg/kg B[a]P exposure groups had significantly higher expression levels of Caspase-3 and Caspase-6 than the blank control group, solvent control group, and 1.0 mg/kg B[a]P exposure group (P < 0.05). The activities of Caspase-3, Caspase-6, and Caspase-9 were significantly higher in the 2.5 and 6.25 mg/kg B[a]P exposure groups than in the blank control group and solvent control group (P < 0.05). There was a positive correlation between the activities of Caspase-3, Caspase-6, and Caspase-9 and early apoptosis ratio of hippocampal neurons in rats (r = 0.793, P = 0.019; r = 0.886, P = 0.006; r = 0.773, P = 0.025). There were no significant differences in the mRNA expression of Bax, Bcl-2, Caspase-3, Caspase-6, and Caspase-9 among these groups (P > 0.05).</p><p><b>CONCLUSION</b>Subchronic exposure to B[a]P can induce apoptosis of hippocampal neurons; its mechanism may be related to the fact that B[a]P can induce upregulated expression of Bax, inhibit expression of Bcl-2, lead to decrease in Bcl-2/Bax ratio, induce upregulated expression of Caspase-3 and Caspase-6, and cause increase in the activities of Caspase-3, Caspase-6, and Caspase-9.</p>

Effects of iodine deficient and hypothyroid on protein expression of extracellular signal-regulate kinase (ERK1/2) in the cerebellum of rats / 中国地方病学杂志

Objective To study the effects of iodine deficiency and hypothyroidism on protein expression of extracellular signal-regulate kinase(ERK1/2) in the cerebellum of rots. Methods Twenty-eight healthy Wistar rots, female, 60 days old, were randomly divided according to their body weight into control group, iodine deficient group and hypothyroidism groups. Hypothyroidism groups in accordance with drinking water containing propylthiouracil(PTU) were divided into doses of 5 mg/L and 15 mg/L groups, 7 rats in each group. Rats after pregnancy, iodine deficient rats were administered with iodine-deficient diet and hypothyroid rats were administered with PTU in drinking water. Pup's cerebellum in each group were weighed on day 14,21,28 and 42. Cerebellum tissue was observed for cerebellar morphology using silver staining and detected for ERK1/2 protein using immunohistochemistry on day 7,14,21,28 and 42. Results On day 14,21,28 and 42, cerebellum weight of pups from iodine-deficient[(0.0945±0.0233), (0.1347±0.0046), (0.1542±0.0094), (0.1949±0.0048)g]were significantly lighter than control[(0.1856±0.0123), (0.2049±0.0098), (0.2268±0.0065), (0.2606±0.0086)g, all P < 0.05]and hypothyroidism groups [for 5 mg/L group: (0.1741±0.0172), (0.1927±0.0103), (0.2181±0.0064), (0.2583±0.0054)g, all P<0.05; for 15 mg/L group: (0.1604±0.0083), (0.1682±0.0103), (0.1996±0.0073) and (0.2579±0.0067)g, all P< 0.05]the difference had statistical significance(F=6.325,8.870, 16.191 and 21.574, all P<0.05). Compared to the controls on day 7, iodine-deficient group didn't have clear layers; on day 21, disappearance of external granule cells from iodine-deficient and 15 mg/L groups was delayed, still two or three layer external granule cells remained; on day 28 and 42, molecular layer from 5, 15 mg/L and iodine-deficient groups and became thinner. Immunohistochemistry showed that on day 7, there was no statistical difference of integrated optical density average of ERK1/2, in all the groups(F=1.102, P>0.05); on day 14,21,28 and 42, integrated optical density average of ERK1/2 in iodine-deficient group[(7.3245±0.5070), (8.3606±1.0683), (9.1217±1.0402), (12.1587±0.7581), all P<0.01]and hypothyroidism groups [for 5 mg/L group: (11.4307±1.5200), (14.919±0.8497), (16.0082±1.1130), (15.7721±0.8293), all P< 0.01; for 15 mg/L group: (7.8538±0.9775), (11.2461±0.8138),(12.78±1.3783), (13.0871±1.1450), all P < 0.01]was significantly lower than those of controls [(16.2831±0.5143), (20.2653±0.9551), (22.7485±1.0267), (22.1725±0.9939), all P < 0.01], the difference having a statistical signifieance(F=16.131,13.543,26.953,41.583, all P<0.01). Conclusions Iodine deficiency and hypothyroidism during critical periods of brain development may change eerebellar morphology and down regulate the protein expression of ERK1/2, which may result in damage of cerebellum development.

Association of INF-gamma/A+874T gene polymorphisms with respiratory syncytial virus bronchiolitis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>A deficient interferon-gamma (IFN-gamma) response has been involved in the pathogenesis of severe respiratory syncytial virus (RSV) infection. Gene polymorphisms in IFN-gamma/A+874T have been associated with the susceptibility to asthma and might be related to disease severity of RSV infection. This study investigated the single nucleotide polymorphisms (SNPs) of IFN-gamma/A+874T in Han children in Wenzhou area and to explore the correlation between gene polymorphisms of IFN-gamma/A+874T and the susceptibility and disease severity of RSV bronchiolitis, as well as the effect of SNPs upon nasopharyngeal secretions (NPS) IFN-gamma and total serum IgE levels.</p><p><b>METHODS</b>One hundred and fourteen hospitalized children with RSV bronchiolitis and 90 healthy controls were recruited. Sequence analysis was used for detecting the SNPs of IFN-gamma/A+874T. NPS IFN-gamma levels were measured using ELISA. Total serum IgE levels were assayed using the chemiluminescence method.</p><p><b>RESULTS</b>IFN-gamma/A+874T gene polymorphisms were present in both the patient and the control groups. AA and AT genotypes were found in both groups, with a AA frequency of 82.5% vs 77.8% and a AT frequency of 17.5% vs 21.1% (p>0.05). The frequency of allele was 90.4% (A) and 9.6% (T) in the patient group, and 88.3% (A) and 11.7% (T) in the control group, respectively. There were no significant differences in the allele frequency between the two groups. Moreover, no difference was found both in NPS IFN-gamma and total serum IgE levels between AA and AT genotypes in the patient group. There were no significant differences in the variation of IFN-gamma/+874 between mild and moderate to severe cases.</p><p><b>CONCLUSIONS</b>IFN-gamma/A+874T gene polymorphisms were present in Han children in Wenzhou area. Gene variations were not associated with the susceptibility and disease severity of RSV bronchiolitis as well as IFN-gamma and total serum IgE levels.</p>