Jing Gu

Metformin, a commonly used drug in the treatment of type II diabetes, may reduce cancer risk and improve cancer prognosis. We evaluated its effect on epithelial ovarian cancer cell lines.The OVCAR-3 and OVCAR-4 cell lines were exposed to metformin with and without cisplatin. Cytotoxicity assays were performed in triplicates using the Alamar colorimetric assay. Levels of total and phosphorylated AMPK, p70S6K and S6K were evaluated by Western blotting following exposure to metformin.Metformin induces dose- and time-dependent growth inhibition of OVCAR-3 and OVCAR-4 cell lines. Metformin potentiated the effect of cisplatin in vitro. Metformin growth inhibition was partly abolished by the AMPK inhibitor, compound C. Western blotting demonstrated that metformin at cytotoxic concentrations, induced AMPK phosphorylation and decreased p70S6K and S6K phosphorylation, suggesting the mechanism for its anti-proliferative action.Metformin significantly inhibits the growth of ovarian cancer cell lines and potentiates cisplatin. Further pre-clinical studies are being conducted to determine the applicability of metformin in the treatment of ovarian cancer.

Introduction The aim of the study was to compare the outcomes of regenerative endodontic treatment (RET) and apexification on immature permanent teeth with pulp necrosis and apical periodontitis. Methods A total of 118 patients (118 teeth) were recruited and randomly assigned to either RET or apexification treatment. Each treatment group was divided into 2 subgroups according to the etiology: dens evaginatus or trauma. Clinical symptoms and complications were recorded, and cone-beam computed tomographic imaging with a limited field of view was used to measure the change of root length, root thickness, and apical foramen size at the 12-month follow-up. The t test/rank sum test and Fisher exact test were applied to compare the change of root morphology between RET and apexification. Results One hundred three of 118 cases were completed at the 12-month follow-up. The survival rate was 100% for both treatment groups. All cases were asymptomatic with apical healing. The RET group showed a significant increase in root length and root thickness compared with the apexification group (P < .05). In the RET group, the cases caused by dens evaginatus achieved increased root length and root thickness compared with those caused by trauma (P < .05). Conclusions RET and apexification achieved a comparable outcome in regard to the resolution of symptoms and apical healing. RET showed a better outcome than apexification regarding increased root thickness and root length. The etiology had an impact on the outcome of RET. Dens evaginatus cases showed better prognoses than trauma cases after RET.

Objectives. Preclinical evaluation of the anti-neoplastic activity of an insulin-like growth factor I receptor (IGF-IR) kinase inhibitor in ovarian cancer. Methods. The OVCAR-3 and OVCAR-4 cell lines were investigated under serum-free tissue culture conditions. IGF-I and IGF-II production were evaluated by standard ELISA and immunohistochemistry. IGF-IR expression and protein levels were evaluated by Western blotting. Cytotoxicity assays were performed in triplicates using the Alamar colorimetric assay. Apoptosis was evaluated by flow cytometry and by Western blotting for PARP. Results. The OVCAR-3 and OVCAR-4 cell lines produce IGF-I and IGF-II, and express IGF-IR, detectable by Western blotting, supporting the existence of an autocrine loop. The existence of this loop justified studies of NVP-AEW541, a small molecular weight inhibitor of the IGF-IR kinase. We observed growth inhibition of the ovarian cancer cell lines, with IC50 between 5 and 15 μM. We also observed that NVP-AEW541 sensitized cells to cisplatin in vitro. Western blotting demonstrated that NVP-AEW541 induced apoptosis at the concentrations that were used in the cytotoxicity assays, and decreased the concentration of the phosphorylated AKT signaling protein downstream of the IGF-IR. Conclusions. IGF-IR is a potential new molecular target in ovarian cancer. The anti-neoplastic activity of NVP-AEW541 in ovarian cancer was observed at concentrations higher than those previously reported for multiple myeloma, suggesting the possibility that a portion of the observed anti-neoplastic activity could involve targets other than the IGF-IR. Experiments are being conducted to investigate the cytotoxicity profile in vivo and the clinical relevance of NVP-AEW541 in ovarian cancer treatment.

As a gene associated with anemia, the erythropoiesis gene is physiologically expressed under hypoxia regulated by †hypoxia-inducing factor-α (HIF-α). Thus, stabilizing HIF-α is a potent strategy to stimulate the expression and secretion of erythropoiesis. In this study, we applied click chemistry to the discovery of HIF prolyl hydroxylase 2 (HIF-PHD2) inhibitors for the first time, and a series of triazole compounds showed preferable inhibitory activity in fluorescence polarization assays. Of particular note was the orally active HIF-PHD inhibitor 15i (IC50 = 62.23 nM), which was almost ten times more active than the phase III drug FG-4592 (IC50 = 591.4 nM). Furthermore, it can upregulate the hemoglobin of cisplatin-induced anemia mice (120 g/L) to normal levels (160 g/L) with no apparent toxicity observed in vivo. These results confirm that triazole compound 15i is a promising candidate for the treatment of renal anemia.

Lysosomes have long been regarded as the “garbage dump” of the cell. More recently, however, researchers have revealed novel roles for lysosomal membranes in autophagy, ion transport, nutrition sensing, and membrane fusion and repair. With active research into lysosomal membrane proteins (LMP), increasing evidence has become available showing that LMPs are inextricably linked to glucose and lipid metabolism, and this relationship represents mutual influence and regulation. In this review, we summarize the roles of LMPs in relation to glucose and lipid metabolism, and describe their roles in glucose transport, glycolysis, cholesterol transport, and lipophagy. The role of transport proteins can be traced back to the original discoveries of GLUT8, NPC1, and NPC2, which were all found to have significant roles in the pathways involved in glucose and lipid metabolism. CLC-5 and SIDT2-knockout animals show serious phenotypic disorders of metabolism, and V-ATPase and LAMP-2 have been found to interact with proteins related to glucose and lipid metabolism. These findings all emphasize the critical role of LMPs in glycolipid metabolism and help to strengthen our understanding of the independent and close relationship between LMPs and glycolipid metabolism.

The regulation and homeostasis of autophagy are essential for maintaining organ morphology and function. As a lysosomal membrane protein, the effect of Sidt2 on kidney structure and renal autophagy is still unknown. In this study, we found that the kidneys of Sidt2-/- mice showed changes in basement membrane thickening, foot process fusion, and mitochondrial swelling, suggesting that the structure of the kidney was damaged. Increased urine protein at 24 h indicated that the kidney function was also damaged. At the same time, the absence of Sidt2 caused a decrease in the number of acidic lysosomes, a decrease in acid hydrolase activity and expression in the lysosome, and an increase of pH in the lysosome, suggesting that lysosomal function was impaired after Sidt2 deletion. The accumulation of autophagolysosomes, increased LC3-II and P62 protein levels, and decreased P62 mRNA levels indicated that the absence of the Sidt2 gene caused abnormal autophagy pathway flow. Chloroquine experiment, immunofluorescence autophagosome, and lysosome fusion assay, and Ad-mcherry-GFP-LC3B further indicated that, after Sidt2 deletion, the production of autophagosomes did not increase, but the fusion of autophagosomes and lysosomes and the degradation of autophagolysosomes were impaired. When incubating Sidt2-/- cells with the autophagy activator rapamycin, we found that it could activate autophagy, which manifested as an increase in autophagosomes, but it could not improve autophagolysosome degradation. Meanwhile, it further illustrated that the Sidt2 gene plays an important role in the smooth progress of autophagolysosome processes. In summary, the absence of the Sidt2 gene caused impaired lysosome function and a decreased number of acidic lysosomes, leading to formation and degradation disorders of the autophagolysosomes, which eventually manifested as abnormal kidney structure and function. Sidt2 is essential in maintaining the normal function of the lysosomes and the physiological stability of the kidneys.

A new series of ortho-naphthoquinone analogs of β-lapachone were designed, synthesized and evaluated. The biological results indicated that most of our compounds were efficient substrates for NQO1. The new scaffold with water-soluble side chain resulted in greater solubility under acidic condition compared to β-lapachone. Thus avoiding the use of hydroxylpropyl β-cyclodextrin which would finally cause the rapid drug clearance from the blood and dose-limiting toxicity in the form of hemolytic anemia. The most soluble and promising compound in this series was 2-((4-benzylpiperazin-1-yl)methyl)naphtho[2,1-d]oxazole-4,5-dione (3k), which inhibited cancer cell (NQO1-rich A549 cell line) growth at IC50 values of 4.6 ± 1.0 μmol·L−1. Furthermore, compound 3k had in vivo antitumor activity in an A549 tumor xenografts mouse model comparable to the activity obtained with β-lapachone. The results indicated that these ortho-naphthoquinones could serve as promising leads for further optimization as novel substrates for NQO1.

Uridine diphosphate-glucuronosyltransferase (UGT) 2B7 is expressed mostly in the human liver, lung and kidney and can transfer endogenous glucuronide group into its substrate and impact the pharmacological effects of several drugs such as estriol, AZT and morphine. UGT2B7 and its allelic variants can dimerize with the homologous enzymes UGT1A1 and UGT1A9, as well as their allelic variants, and then change their enzymatic activities in the process of substrate catalysis. The current study was designed to identify this mechanism using morphine as the substrate of UGT2B7. Single-recombinant allozymes, including UGT2B7*1 (wild type), UGT2B7*71S (A71S, 211G>T), UGT2B7*2 (H268Y, 802C>T), UGT2B7*5 (D398N, 1192G>A), and double-recombinant allozymes formed by the dimerization of UGT1A9*1 (wild type), UGT1A9*2 (C3Y, 8G>A), UGT1A9*3 (M33T, 98T>C), UGT1A9*5 (D256N, 766G>A), UGT1A1 (wild type) with its splice variant UGT1A1b were established and incubated with morphine in vitro. Each sample was analyzed with HPLC-MS/MS. All enzyme kinetic parameters were then measured and analyzed. From the results, the production ratio of its aberrant metabolism and subsequent metabolites, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G), changes regioselectively. Double-recombinant allozymes exhibit stronger enzymatic activity catalyzing morphine than the single-recombinant alloyzymes. Compared to UGT2B7*1, UGT2B7*2 singles or doubles have lower Km values for M3G and M6G, whereas UGT2B7*5 allozymes perform opposite effects. The double allozymes of UGT1A9*2 or UGT1A9*5 with UGT2B7 tend to produce M6G. Interestingly, the majority of single or double allozymes significantly reduce the ratio of M3G to M6G. The UGT1A9*2-UGT2B7*1 double enzyme has the lowest M3G:M6G ratio, reflecting that more M6G would form in morphine glucuronide metabolism. This study demonstrates that UGT2B7 common SNPs and their dimers with UGT1A1 and UGT1A9 and their allelic variants can regioselectively affect the generation of two metabolites of morphine via altering the CLint ratios of M3G to M6G. These results may predict the effectiveness of morphine antinociception in individualized opioid treatment.

Abstract Regulating main brain-uptake transporter of morphine may restrict its tolerance generation, then modify its antinociception. In this study, more than 2 fold higher intracellular uptake concentrations for morphine and morphine-6-glucuronide (M6G) were observed in stable expression cells, HEK293-hOATP2B1 than HEK293-MOCK. Specifically, the K m value of morphine to OATP2B1 (57.58 ± 8.90 μM) is 1.4-time more than that of M6G (80.31 ± 21.75 μM); Cyclosporine A (CsA), an inhibitor of OATP2B1, can inhibit their intracellular accumulations with IC 50 = 3.90 ± 0.50 μM for morphine and IC 50 = 6.04 ± 0.86 μM for M6G, respectively. To further investigate the role of OATP2B1 in morphine brain transport and tolerance, the novel nanoparticles of DGL-PEG/dermorphin capsulated siRNA (OATP2B1) were applied to deliver siRNA into mouse brain. Along with OATP2B1 depressed, a main reduction was found for each of morphine or M6G in cerebrums or epencephalons of acute morphine tolerance mice. Furthermore, calcium/calmodulin-dependent protein kinase IIα (CaMKIIα) in mouse prefrontal cortex (mPFC) underwent dephosphorylation at Thr286. In conclusion, OATP2B1 downregulation in mouse brain can suppress tolerance via blocking morphine and M6G brain transport. These findings might help to improve the pharmacological effects of morphine.

Precise public health and clinical interventions for the COVID-19 pandemic has spurred a global rush on SARS-CoV-2 variant tracking, but current approaches to variant tracking are challenged by the flood of viral genome sequences leading to a loss of timeliness, accuracy, and reliability. Here, we devised a new co-mutation network framework, aiming to tackle these difficulties in variant surveillance.To avoid simultaneous input and modeling of the whole large-scale data, we dynamically investigate the nucleotide covarying pattern of weekly sequences. The community detection algorithm is applied to a co-occurring genomic alteration network constructed from mutation corpora of weekly collected data. Co-mutation communities are identified, extracted, and characterized as variant markers. They contribute to the creation and weekly updates of a community-based variant dictionary tree representing SARS-CoV-2 evolution, where highly similar ones between weeks have been merged to represent the same variants. Emerging communities imply the presence of novel viral variants or new branches of existing variants. This process was benchmarked with worldwide GISAID data and validated using national level data from six COVID-19 hotspot countries.A total of 235 co-mutation communities were identified after a 120 weeks' investigation of worldwide sequence data, from March 2020 to mid-June 2022. The dictionary tree progressively developed from these communities perfectly recorded the time course of SARS-CoV-2 branching, coinciding with GISAID clades. The time-varying prevalence of these communities in the viral population showed a good match with the emergence and circulation of the variants they represented. All these benchmark results not only exhibited the methodology features but also demonstrated high efficiency in detection of the pandemic variants. When it was applied to regional variant surveillance, our method displayed significantly earlier identification of feature communities of major WHO-named SARS-CoV-2 variants in contrast with Pangolin's monitoring.An efficient genomic surveillance framework built from weekly co-mutation networks and a dynamic community-based variant dictionary tree enables early detection and continuous investigation of SARS-CoV-2 variants overcoming genomic data flood, aiding in the response to the COVID-19 pandemic.

Isoform expression of CD44 in follicular carcinoma (FC) of the thyroid was analyzed by Immunohistochemlcal staining and compared to the isoforms In follicular adenoma (FA) and papillary carcinoma (PC) of the thyroid. Variant isoforms of CD44 (CD44v) were detected In these neoplastic cells but not In non‐neoplastlc cells. CD44v6 was expressed In PC with nodal metastasis and also in FC at significantly higher frequencies than those in PC without metastasis and FA. The frequency of expression of CD44v3 was significantly higher In PC with nodal metastasis than in PC without metastasis. The reverse transcription‐polymerase chain reaction (RT‐PCR) followed by Southern blotting analysis revealed the presence of a transcript for a variant of CD44 that contained variant exon 6 in FA, FC and PC. DNA sequencing of the products of RT‐PCR yielded three species of cDNA for CD44v. One of the cDNA corresponded to a transcript that contained variant exon 6. These results suggest that Immunohistochemlcal staining and RT‐PCR with Southern blotting analysis for CD44v6 might be a useful diagnostic tool for the differentiation of FC from FA and that the expression of CD44v3 and CD44v6 might be important for the development of nodal metastasis in cases of PC.

In this study, we explored the role and mechanisms of Cyclocarya paliurus polysaccharide on cell apoptosis in thyroid cancer (TC) cells. The apoptosis of thyroid cancer cells in vitro and tumor tissues in vivo induced by Cyclocarya paliurus polysaccharide was determined by MTT assay and flow cytometric assay. The downstream molecules including phosphop-protein kinase B (p-Akt), Akt, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) in tumor tissue were evaluated by western blotting. MTT and flow cytometry assay in vitro revealed Cyclocarya paliurus polysaccharide-induced apoptosis of thyroid cancer cell line in a manner of time-dependent and dose-dependent. In vivo assay showed 50 mg/kg and 100 mg/kg Cyclocarya paliurus polysaccharide significantly suppressed the proliferation of thyroid cancer in mice. Western blotting showed downregulation of p-Akt, Akt, and Bcl-2 and upregulation of Bax. These results suggest that Cyclocarya paliurus polysaccharide may enhance thyroid cancer cell apoptosis by suppressing the activation of p-Akt, Akt, and Bcl-2 and activating Bax, which provide a novel use of CPP as a thyroid cancer treatment.

Uridine diphosphate-glucuronosyltransferase (UGT) 2B7, as one of significant drug enzymes, is responsible on the glucuronidation of abundant endobiotics or xenobiotics.We here report that it is markedly repressed in the tumor tissues of colorectal carcinoma (CRC) patients.Accordingly, morphine in CRC cells will stimulate the expression of its main metabolic enzyme, UGT2B7 during tolerance generation by activating the positive signals in histone 3, especially for trimethylated lysine 27 (H3K4Me3) and acetylated lysine 4 (H3K27Ac).Further study reveals that brainderived neutrophilic factor (BDNF), a secretory neurotrophin, enriched in CRC can interact and inhibit UGT2B7 by primarily blocking the positive signals of H3K4Me3 as well as activating H3K27Ac on the promoter region of UGT2B7.Meanwhile, BDNF repression attributes to the sensitizations of main core factors in poly-comb repressive complex (PRC) 1 rather than PRC2 as the reason of the depression of SUZ12 in the later complex.Besides that, the productions of two main morphine glucuronides are both increased in the BDNF deficient or TSA and BIX-01294 treated morphine tolerance-like HCT-116 cells.On the same condition, active metabolite, morphine-6glucuronide (M6G) was accumulated more than inactive M3G.Our findings imply that enzymatic activity enhancement and substrate regioselective catalysis alteration of UGT2B7 may release morphine tolerance under the cure of tumor-induced pain.

Diabetic nephropathy (DN) is one of the most lethal complications of diabetes mellitus with chronic inflammation. We have examined the role of the inflammatory chemokine CCL24 in DN. We observed that serum levels of CCL24 were significantly elevated in patients with DN. Not only that, the expression of CCL24 was significantly increased in the kidneys of DN mice. The kidney of DN mice showed increased renal fibrosis and inflammation. We characterized an in vitro podocyte cell model with high glucose. Western blot analysis showed that expression of CCL24 was significantly increased under high-glucose conditions. Stimulation with high glucose (35 mmol/L) resulted in an increase in CCL24 expression in the first 48 hours but changed little after 72 hours. Moreover, with glucose stimulation, the level of podocyte fibrosis gradually increased, the expression of the proinflammatory cytokine IL-1β was upregulated, and the expression of the glucose transporter GLUT4, involved in the insulin signal regulation pathway, also increased. It is suggested that CCL24 is involved in the pathogenesis of DN. In order to study the specific role of CCL24 in this process, we used the CRISPR-Cas9 technique to knock out CCL24 expression in podocytes. Compared with the control group, the podocyte inflammatory response induced by high glucose after CCL24 knockout was significantly increased. These results suggest that CCL24 plays a role in the development of early DN by exerting an anti-inflammatory effect, at least, in podocytes.

The SID1 transmembrane family, member 2, namely, Sidt2, is a highly glycosylated multichannel lysosomal transmembrane protein, but its specific physiological function remains unknown. Lysosomal membrane proteins are very important for the executive functioning of lysosomes. As an important part of the lysosomal membrane, Sidt2 can maintain the normal morphology of lysosomes and help stabilize them from the acidic pH environment within. As a receptor/transporter, it binds and transports nucleic acids and mediates the uptake and degradation of RNA and DNA by the lysosome. During glucose metabolism, deletion of Sidt2 can cause an increase in fasting blood glucose and the impairment of grape tolerance, which is closely related to the secretion of insulin. During lipid metabolism, the loss of Sidt2 can cause hepatic steatosis and lipid metabolism disorders and can also play a role in signal regulation and transport. Here, we review the function of the lysosomal membrane protein Sidt2, and focus on its role in glucose and lipid metabolism, autophagy and nucleotide (DNA/RNA) transport.

Objective To assess the psychological status of staff at the centers for disease control and prevention(CDC) in Sichuan during the outbreak of coronavirus disease 2019(COVID-19) and explore the influencing factors. Methods The staff at Sichuan provincial,municipal,and county(district)-level CDC were selected by convenience sampling.Their basic information,work status,training status,work difficulties,and support from the work group were collected from the self-filled questionnaires online.The Generalized Anxiety Disorder Scale and the 9-question Patient Health Questionnaire were respectively employed to measure the anxiety and depression of the staff.The stepwise Logistic regression was carried out to analyze the influencing factors of anxiety and depression in CDC staff. Results Among the 653 staff,58.35% and 50.06% presented anxiety and depression,respectively.The regression results showed that age(OR=0.95,95%CI=0.92-0.97) and mental support from the work group(OR=0.61,95%CI=0.45-0.82) were the protective factors while physical fatigue(OR=1.82,95%CI=1.20-2.74),work pressure(OR=1.61,95%CI=1.21-2.12),and insufficient protective equipment(OR=1.92,95%CI=1.06-3.49) were the risk factors for depression of CDC staff.Age(OR=0.97,95%CI=0.94-0.99),length of sleep per day(OR=0.74,95%CI=0.56-0.96),and participation in technical training(OR=0.33,95%CI=0.12-0.95) were the protective factors while mental fatigue(OR=1.68,95%CI=1.18-2.41),work pressure(OR=2.94,95%CI=2.08-4.17),and unclear incentive system for overtime(OR=1.99,95%CI=1.23-3.23) were the risk factors for the anxiety of CDC staff. Conclusion The anxiety and depression status of CDC staff during the COVID-19 outbreak were worrying,which were mainly affected by age,sleep,supply of protective equipment,incentive system,fatigue,and work pressure.

Evidence on the interaction of lifestyle and long-term ambient particle (PM) exposure on the prevalence of hypertension, diabetes, particularly their combined condition is limited. We investigate the associations between PM and these outcomes and whether the associations were modified by various lifestyles.This was a large population-based survey during 2019-2021 in Southern China. The concentrations of PM were interpolated and assigned to participants by the residential address. Hypertension and diabetes status were from questionnaires and confirmed with the community health centres. Logistic regression was applied to examine the associations, followed by a comprehensive set of stratified analyses by the lifestyles including diet, smoking, drinking, sleeping and exercise.A total of 82 345 residents were included in the final analyses. For each 1 μg/m3 increase in PM2.5, the adjusted OR for the prevalence of hypertension, diabetes and their combined condition were 1.05 (95% CI 1.05 to 1.06), 1.07 (95% CI 1.06 to 1.08) and 1.05 (95% CI 1.04 to 1.06), respectively. We observed that the association between PM2.5 and the combined condition was greatest in the group with 4-8 unhealthy lifestyles (OR=1.09, 95% CI 1.06 to 1.13) followed by the group with 2-3 and those with 0-1 unhealthy lifestyle (P interaction=0.026). Similar results and trends were observed in PM10 and/or in those with hypertension or diabetes. Individuals who consumed alcohol, had inadequate sleep duration or had poor quality sleep were more vulnerable.Long-term PM exposure was associated with increased prevalence of hypertension, diabetes and their combined condition, and those with unhealthy lifestyles suffered greater risks of these conditions.

As a self-degrading and highly conserved survival mechanism, autophagy plays an important role in maintaining cell survival and recycling. The discovery of autophagy-related (ATG) genes has revolutionized our understanding of autophagy. Lysosomal membrane proteins (LMPs) are important executors of lysosomal function, and increasing evidence has demonstrated their role in the induction and regulation of autophagy. In addition, the functional dysregulation of the process mediated by LMPs at all stages of autophagy is closely related to neurodegenerative diseases and cancer. Here, we review the role of LMPs in autophagy, focusing on their roles in vesicle nucleation, vesicle elongation and completion, the fusion of autophagosomes and lysosomes, and degradation, as well as their broad association with related diseases.

Current global health course is most set as elective course taught in traditional teacher-taught model with low credit and short term. Innovate teaching models are required. Crowdsourcing characterized by high flexibility and strong application-orientation holds its potential to enhance global health education. We applied crowdsourcing to global health teaching for undergraduates, aiming to develop and evaluate a new teaching model for global health education.Crowdsourcing was implemented into traditional course-based teaching via introducing five COVID-19 related global health debates. Undergraduate students majoring in preventative medicine and nursing grouped in teams of 5-8, were asked to resolve these debates in reference to main content of the course and with manner they thought most effective to deliver the messages. Students' experience and teaching effect, were evaluated by questionnaires and teachers' ratings, respectively. McNemar's test was used to compare the difference in students' experience before and after the course, and regression models were used to explore the influencing factors of the teaching effect.A total of 172 undergraduates were included, of which 122 (71%) were females. Students' evaluation of the new teaching model improved after the course, but were polarized. Students' self-reported teaching effect averaged 67.53 ± 16.8 and the teachers' rating score averaged 90.84 ± 4.9. Students majoring in preventive medicine, participated in student union, spent more time on revision, and had positive feedback on the new teaching model tended to perform better.We innovatively implemented crowdsourcing into global health teaching, and found this new teaching model was positively received by undergraduate students with improved teaching effects. More studies are needed to optimize the implementation of crowdsourcing alike new methods into global health education, to enrich global health teaching models.

Alzheimer's disease (AD) is characterized by β-amyloid (Aβ) overproduction and tau hyperphosphorylation. Familial Alzheimer disease mutations of presenilin 1 (PS-1) enhance the generation of Aβ42 and intracellular accumulation. However, the effect of PS-1 mutations on tau phosphorylation and the underlying mechanisms were poorly understood, and there is no treatment to arrest tau pathology. Western blot, Immunohistochemistry and SOD activity assay were performed in this study. Tau phosphorylation at Ser396, Ser404 and Tau-1 (Ser198/199/202) epitopes was increased in hippocampus of the aged transgenic mice overexpressing Alzheimer presenilin 1-L235P(leucine-to-proline mutation at codon 235, 16–17 months old) compared with the age-matched wild type mice. Meanwhile, we further detected the alterations in Akt-glycogen synthase kinase (GSK)–3β signaling associated with oxidative stress and found decreased phosphorylation of both Akt and GSK-3β in the transgenic mice. Concurrently, we detected an increased oxidative stress reaction as evidenced by elevated level of malondialdehyde (MDA) and decreased activity of superoxide dismutase (SOD) in hippocampus of the transgenic mice. In addition, exogenous supplementation of antioxidant coenzyme Q10 (CoQ10) not only partially decreased MDA level and up-regulated the activity of SOD, but also attenuated tau hyperphosphorylation, concurrently accompanied with a restoration of Akt-GSK-3β signaling. These results suggest that tau hyperphosphorylation is associated with enhanced oxidative stress and the mechanisms may involve oxidative stress-mediated depression of Akt-GSK-3β signaling. The neuroprotective effect of CoQ10 could be attributed to its antioxidant property. Our data indicate that CoQ10 might be a potential therapeutic candidate in AD treatment.

Objective To measure the expression of protein kinase D1 (PKD1) , tyr463-phos-phorylaed PKD1 (pPKD1-tyr463) and ser916-phos-phorylaed PKD1 (pPKD1-ser916) in squamous cell carcinoma (SCC) , Bowen's disease (BD) and actinic keratosis (AK) , and to explore their significance. Methods Fresh tissue samples were resected from lesions of patients with SCC (SCC group) , BD (BD group) and AK (AK group) , as well as from normal skin of healthy human controls (control group) , and each group had a sample size of 10. Real-time RT-PCR was performed to measure the mRNA expression of protein kinase D1 gene (PRKD1) , and Western blot analysis to determine the protein expression of PKD1, pPKD1-tyr463 and pPKD1-ser916. In addition, immunohistochemical study was conducted to determine the expression of PKD1, pPKD1-tyr463 and pPKD1-ser916 in another 50 paraffin-embedded skin samples of SCC, 20 samples of BD, 20 samples of AK and 10 normal skin samples. Results PRKD1 mRNA expression significantly differed among the control group (0.64 ± 0.09) , SCC group (5.37 ± 1.06) , BD group (2.69 ± 0.72) and AK group (2.43 ± 0.46) (F= 21.37, P 0.05) . Immunohistochemical study showed that the total PKD1 protein and pPKD1-tyr463 in the SCC and BD groups were mainly expressed in the cytoplasm and cell membrane of spinous layer cells and atypical cells, and their expression rates were significantly higher than those in the AK group and control group (all P < 0.01) . The pPKD1-ser916 was only slightly expressed in some cancer nests of well-differentiated SCC tissues, but not in poorly-differentiated SCC, AK, BD tissues and normal skin tissues. In the SCC group, the expression rate of PKD1 increased with the increase of the pathological grade of SCC, and the PKD1 expression was positively correlated with pPKD1-tyr463 expression (rc c= 0.479, P < 0.05) . Western blot results were consistent with immunohistochemical findings. Conclusion PKD1 and pPKD1-tyr463 may be involved in the development and differentiation of skin tumors derived from stratified squamous epithelium, and PKD1 may exert promotive effects on the formation of cutaneous SCC by activating the Tyr463 phosphorylation site. Key words: Carcinoma, squamous cell; Bowen's disease; Keratosis, actinic; Protein kinase D1

Abstract β-cell dedifferentiation is important in the pathogenesis of type 2 diabetes, and the relationship between lysosome membrane proteins and pancreatic β-cell dedifferentiation is still unknown. Sidt2 is a lysosomal membrane protein identified in our previous study. After Sidt2 elimination, mice exhibited a diabetic phenotype. In the present study, we found that Sidt2 also can be considered a new insulin granule membrane protein. Sidt2 deficiency resulted in islet dysfunction and inhibited insulin secretion. The absence of Sidt2 also led to increased β cell dedifferentiation in mice. Further experiments showed that defects in insulin secretion caused dedifferentiation in the absence of Sid2. In summary, our study has found a new dedifferentiation regulatory mechanism mediated by a lysosomal membrane protein. This new secretory regulation pathway will enrich our understanding of the regulatory mechanisms involved in β cell dedifferentiation and provide new insights into the pathogenesis of diabetes.

Objective To explore the efficacy of salmon calcitonin combined with vitamin D3 in the treatment of chronic renal insufficiency and secondary hyperthyroidism and its effects on bone calcium metabolism. Methods A total of 40 patients with chronic renal failure who underwent hemodialysis in our blood purification center from September 2017 to December 2017 were selected for the study and were divided into treatment group (20 cases) and control group (20 cases) according to the random number table method. Patients in control group were given conventional treatment + calcitriol orally, and patients in treatment group were given subcutaneous injection of salmon calcitonin on the basis of control group. The course of treatment in both groups was 12 weeks. The clinical efficacy and clinical symptoms improvement, serum calcium, serum phosphorus and parathyroid hormone (iPTH) as well as the adverse reactions were compared between the two groups. The Kidney Disease Targeted Areas (KDTA) and Visual Analogue Pain Score (VAS) were used to evaluate the patients' quality of life and bone pain. Results The effective rate in treatment group was significantly higher than that in control group (95.00% vs 65.00%) (P 0.05). Conclusions The treatment effect of salmon calcitonin combined with vitamin D3 on chronic renal insufficiency secondary hyperthyroidism is obvious, and the clinical symptoms such as bone pain are improved obviously. It is believed that salmon calcitonin is related to the regulation of calcium, phosphorus and bone metabolism. Key words: Renal insufficiency, chronic; Hyperparathyroidism, secondary; Salmon calcitonin; Vitamin D3

Abstract Background: To fight against COVID-19, many policymakers are wavering on stricter public health interventions. However, relying on these measures but different strategies, both in and out of China’s Hubei province basically contained the epidemic in late February 2020. This study aimed to assess the response process and estimate time-varying effect of Hubei control strategy to provide insights for intervention design and implementation. Methods: We retrospectively compared the spread and control of COVID-19 between China’s Hubei (excluding Wuhan) and non-Hubei areas using data that includes case reports, human mobility, and public health interventions from 1 January to 29 February, 2020. The static and dynamic risk assessment models were developed to statistically investigate the effect trends of Hubei control strategy on case growth after adjusting importation risk and response timing with non-Hubei strategy as a contrast. Results: The analysis detected much higher but differential importation risk in Hubei. The response timing largely coincided with the importation risk in non-Hubei areas, but Hubei areas showed an opposite pattern. A careful and comprehensive comparison showed that Hubei control strategy implemented interventions characterized by unprecedentedly strict and ‘monitored’ self-quarantine at home, while non-Hubei strategy included physical distancing measures to reduce contact among individuals within or between populations. In contrast with non-Hubei control strategy, Hubei strategy showed a much higher, non-linear and gradually diminishing protective effect with at least 3 times fewer cases. Conclusions: A risk-based control strategy is crucial to design an effective response for COVID-19 control. Our study demonstrates that the stricter Hubei strategy can achieve much better control effectiveness. These findings highlight the health benefits of precise and differentiated strategies informed by constant monitoring of outbreak risk and policy impacts.