[Zinc finger protein-36 deficiency inhibits osteogenic differentiation of mouse bone marrow-derived mesenchymal stem cells and preosteoblasts by activating the ERK/MAPK pathway].

OBJECTIVE: To explore the role of zinc finger protein 36(ZFP36) in regulating osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and preosteoblasts. METHODS: ZFP36 expression was observed in primary mouse BMSCs and mouse preosteoblasts (MC3T3-E1 cells) during induced osteogenic differentiation. Zfp36-deficient cell models were constructed in the two cells using RNA interference technique and the changes in differentiation capacities of the transfected cells into osteoblasts were observed. Transcriptome sequencing was used to investigate the potential mechanisms of ZFP36 for regulating osteoblast differentiation of the two cells. U0126, a ERK/MAPK signal suppressor, was used to verify the regulatory mechanism of Zfp36 in osteogenic differentiation of Zfp36-deficient cells. RESULTS: During the 14-day induction of osteogenic differentiation, both mouse BMSCs and MC3T3-E1 cells exhibited increased expression of ZFP36, and its mRNA expression reached the peak level on Day 7(P < 0.0001). The Zfp36-deficient cell models showed reduced intensity of alkaline phosphatase (ALP) staining and alizarin red staining with significantly lowered expressions of the osteogenic marker genes including Alpl, Sp7, Bglap and Ibsp (P < 0.01). Transcriptome sequencing verified the reduction of bone mineralization-related gene expressions in Zfp36-deficient cells and indicated the involvement of ERK signaling in the potential regulatory mechanism of Zfp36. Immunoblotting showed that pERK protein expression increased significantly in Zfp36-deficient cells compared with the control cells. In Zfp36-deficient MC3T3-E1 cells, inhibition of activated ERK/MAPK signaling with U0126 resulted in obviously enhanced ALP staining and significantly increased expressions of osteoblast differentiation markers Runx2 and Bglap (P < 0.05). CONCLUSIONS: ZFP36 is involved in the regulation of osteoblast differentiation of mouse BMSCs and preosteoblasts, and ZFP36 deficiency causes inhibition of osteoblast differentiation of the cells by activating the ERK/MAPK signaling pathway.

Notch signaling regulates Th17 cells differentiation through PI3K/AKT/mTORC1 pathway and involves in the thyroid injury of autoimmune thyroiditis.

PURPOSE: Autoimmune Thyroiditis (AIT) is the most common thyroid disease; however, there were no measures to prevent the progression of the disease. The present study attempts to identify that Notch signaling regulates the differentiation of T helper 17 (Th17) cells by activating downstream Phosphatidylinositol-3 kinase/protein kinase/mechanistic target of rapamycin complex 1 (PI3K/AKT/mTORC1) pathway participating in the thyroid injury of the experimental autoimmune thyroiditis (EAT). METHODS: In vivo experiments, mice were randomly divided into 4 groups: a control group, an EAT group, and two groups with LY294002 treatment (pTg plus 25 mg/kg or 50 mg/kg LY294002, respectively). The degrees of thyroiditis were evaluated, and the percentage of Th17 cells, expression of interleukin-17A (IL-17A), and the main components of the Notch-PI3K signaling pathway were detected in different groups. In vitro experiments, two different dosages of LY294002 (25 and 50 µM) were used to intervene splenic mononuclear cells (SMCs) from EAT mice to further evaluate the regulatory effect of Notch-PI3K pathway on Th17 cells. RESULTS: Our data demonstrate that the infiltration of Th17 cells and the expressions of IL-17A, Notch, hairy and split 1 (Hes1), p­AKT (Ser473), p­AKT (Thr308), p­mTOR (Ser2448), S6K1, and S6K2 increased remarkably in EAT mice. After PI3K pathway was blocked, the degrees of thyroiditis were significantly alleviated, and the proportion of Th17 cells, the expression of IL-17A, and the above Notch-PI3K pathway-related molecules decreased in a dose-dependent manner. Additionally, the proportion of Th17 cells was positively correlated with the concentration of serum thyroglobulin antibody (TgAb), IL-17A, and Notch-PI3K pathway-related molecules mRNA levels. CONCLUSIONS: Notch signal promotes the secretion of IL-17A from Th17 cells by regulating the downstream PI3K/AKT/mTORC1 pathway through Hes-Phosphatase and tensin homolog (PTEN) and participates in thyroid autoimmune damage, and the PI3K pathway inhibitor may play important effects on AIT by affecting Th17 cells differentiation.

Chemo-Enzymatic Synthesis of Isomeric I-branched Polylactosamines Using Traceless Blocking Groups.

Poly-N-acetyl lactosamines (polyLacNAc) are common structural motifs of N- and O-linked glycan, glycosphingolipids and human milk oligosaccharides. They can be branched by the addition of ß1,6-linked N-acetyl-glucosamine (GlcNAc) moieties to internal galactoside (Gal) residues by the I-branching enzyme beta-1,6-N-acetylglucosaminyltransferase 2 (GCNT2). I-branching has been implicated in many biological processes and is also associated with various diseases such as cancer progression. Currently, there is a lack of methods that can install, in a regioselective manner, I-branches and allows the preparation of isomeric poly-LacNAc derivatives. Here, we described a chemo-enzymatic strategy that addresses this deficiency and is based on the enzymatic assembly of an oligo-LacNAc chain that at specific positions is modified by a GlcNTFA moiety. Replacement of the trifluoroacetyl (TFA) moiety by tert-butyloxycarbonyl (Boc) gives compounds in which the galactoside at the proximal site is blocked from modification by GCNT2. After elaboration of the antennae, the Boc group can be removed, and the resulting amine acetylated to give natural I-branched structures. It is also shown that fucosides can function as a traceless blocking group that can provide complementary I-branched structures from a single precursor. The methodology made it possible to synthesize a library of polyLacNAc chains having various topologies.

COVID-19 vaccine safety and side effects in children aged 5-11 years: a cross-sectional study.

INTRODUCTION: The COVID-19 pandemic has prompted a global drive for vaccination, including children. Despite the urgency, understanding the safety and side effects remains crucial. Our study aimed to evaluate the safety of the Pfizer- BioNTech (BNT162b2) vaccine in children by determining the proportion of vaccinated children who experienced side effects and identifying factors associated with postvaccination side effects. MATERIALS AND METHODS: A cross-sectional study was conducted among children who received the COVID-19 vaccine between 3 February and 8 May 2022. Data were collected using a self-administered questionnaire filled out by the parent or legal guardian. RESULTS: The mean age of the study participants was 9 years old and 43.1% were males. Out of the 195 participants in the study, 62 (31.8%) reported side effects after vaccination. The most frequently reported side effects were pain at the injection site (29.7%, n=58), fever (15.9%, n=31), localised inflammation (10.8%, n=21) and arthralgia/myalgia (9.2%, n=18). There were no reported severe adverse events such as anaphylaxis or myocarditis. Most side effects occurred within the first two days post-vaccination. There was a higher proportion of side effects among children with underlying co-morbidities. No significant differences were observed based on age, weight, ethnicity and the presence of allergies, or the use of premedication. CONCLUSION: The BNT162b2 vaccine was generally welltolerated in children, with most side effects being mild and self-limiting. These findings support the safety of the COVID-19 vaccine and would guide healthcare professionals, parents and policy-makers in making informed decisions about COVID-19 vaccination, especially among high-risk groups.

Sialic acid O-acetylation patterns and glycosidic linkage type determination by ion mobility-mass spectrometry.

O-acetylation is a common modification of sialic acids that has been implicated in a multitude of biological and disease processes. A lack of analytical methods that can determine exact structures of sialic acid variants is a hurdle to determine roles of distinct O-acetylated sialosides. Here, we describe a drift tube ion mobility-mass spectrometry approach that can elucidate exact O-acetylation patterns as well as glycosidic linkage types of sialosides isolated from complex biological samples. It is based on the use of a library of synthetic O-acetylated sialosides to establish intrinsic collision cross section (CCS) values of diagnostic fragment ions. The CCS values were used to characterize O-acetylated sialosides from mucins and N-linked glycans from biologicals as well as equine tracheal and nasal tissues. It uncovered contrasting sialic acid linkage types of acetylated and non-acetylated sialic acids and provided a rationale for sialic acid binding preferences of equine H7 influenza A viruses.

Andrographolide loaded montmorillonite attenuated enterotoxigenic Escherichia coli induced intestinal barrier injury and inflammation in a mouse model.

Montmorillonite (MMT), a natural absorbent agent, has widely been accepted for its antidiarrhea function in human and farm animals; however, its specific physicochemical property limits its biological function in practical use. In the current study, raw MMT was loaded by andrographolide, namely andrographolide loaded montmorillonite (AGP-MMT). The microstructure of AGP-MMT was observed by scanning electron microscope (SEM) and X-ray diffraction (XRD). The effect of AGP-MMT on the growth performance, intestinal barrier and inflammation was investigated in an enterotoxigenic Escherichia coli (ETEC) challenged mice model. The results show that the microstructure of MMT was obviously changed after andrographolide modification: AGP-MMT exhibited a large number of spheroid particles, and floccule aggregates, but lower interplanar spacing compared with MMT. ETEC infection induced body weight losses and intestinal barrier function injury, as indicated by a lower villus height and ratio of villus height/crypt depth, whereas the serum levels of diamine oxidase (DAO), D-xylose and ETEC shedding were higher in the ETEC group compared with the CON group. Mice pretreated with AGP-MMT showed alleviated body weight losses and the intestinal barrier function injury induced by ETEC challenge. The villus height and the ratio of villus height/crypt depth, were higher in mice pretreated with AGP-MMT than those pretreated with equal levels of MMT. Pretreatment with AGP-MMT also alleviated the increased concentration of serum tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß), and the corresponding genes in the jejunum induced by ETEC infection in mice. The protein and mRNA levels of IL-1ß were lower in mice pretreated with AGP-MMT than those with equal levels of MMT. The results indicate that AGP-MMT was more effective in alleviating intestinal barrier injury and inflammation in mice with ETEC challenge than MMT.

Energetic Electron Precipitation Driven by Electromagnetic Ion Cyclotron Waves from ELFIN's Low Altitude Perspective.

We review comprehensive observations of electromagnetic ion cyclotron (EMIC) wave-driven energetic electron precipitation using data collected by the energetic electron detector on the Electron Losses and Fields InvestigatioN (ELFIN) mission, two polar-orbiting low-altitude spinning CubeSats, measuring 50-5000 keV electrons with good pitch-angle and energy resolution. EMIC wave-driven precipitation exhibits a distinct signature in energy-spectrograms of the precipitating-to-trapped flux ratio: peaks at >0.5 MeV which are abrupt (bursty) (lasting ∼17 s, or ΔL∼0.56) with significant substructure (occasionally down to sub-second timescale). We attribute the bursty nature of the precipitation to the spatial extent and structuredness of the wave field at the equator. Multiple ELFIN passes over the same MLT sector allow us to study the spatial and temporal evolution of the EMIC wave - electron interaction region. Case studies employing conjugate ground-based or equatorial observations of the EMIC waves reveal that the energy of moderate and strong precipitation at ELFIN approximately agrees with theoretical expectations for cyclotron resonant interactions in a cold plasma. Using multiple years of ELFIN data uniformly distributed in local time, we assemble a statistical database of ∼50 events of strong EMIC wave-driven precipitation. Most reside at L∼5-7 at dusk, while a smaller subset exists at L∼8-12 at post-midnight. The energies of the peak-precipitation ratio and of the half-peak precipitation ratio (our proxy for the minimum resonance energy) exhibit an L-shell dependence in good agreement with theoretical estimates based on prior statistical observations of EMIC wave power spectra. The precipitation ratio's spectral shape for the most intense events has an exponential falloff away from the peak (i.e., on either side of ∼1.45 MeV). It too agrees well with quasi-linear diffusion theory based on prior statistics of wave spectra. It should be noted though that this diffusive treatment likely includes effects from nonlinear resonant interactions (especially at high energies) and nonresonant effects from sharp wave packet edges (at low energies). Sub-MeV electron precipitation observed concurrently with strong EMIC wave-driven >1 MeV precipitation has a spectral shape that is consistent with efficient pitch-angle scattering down to ∼ 200-300 keV by much less intense higher frequency EMIC waves at dusk (where such waves are most frequent). At ∼100 keV, whistler-mode chorus may be implicated in concurrent precipitation. These results confirm the critical role of EMIC waves in driving relativistic electron losses. Nonlinear effects may abound and require further investigation.

Brain structural and functional MRI alterations and their predictive value for the visual outcome at 3 years in clinically isolated optic neuritis.

AIM: To investigate the structural and functional magnetic resonance imaging (MRI) alterations and their clinical significance for predicting visual outcomes at 3 years in clinically isolated optic neuritis (CION). MATERIALS AND METHODS: Forty-three CION patients and 44 matched healthy controls (HC) underwent a three dimensional (3D) T1-weighted and resting-state functional MRI using a 3 T MRI system. Grey-matter volume (GMV) and functional MRI measures were compared among HC and CION patients with good and poor outcomes. The correlations between MRI measures and visual outcomes were investigated, and a binary logistic regression model was performed to predict the visual outcome. RESULTS: CION patients with good and poor outcomes showed similar patterns of decreased GMV and increased functional MRI activities compared to HC. Compared to patients with good visual recovery, CION patients with poor visual recovery showed significantly reduced GMV in the insula and superior temporal gyrus (STG), decreased amplitude of low-frequency fluctuation (ALFF) in the inferior frontal gyrus (IFG), and increased functional activities in the middle frontal gyrus (MFG) and middle temporal gyrus (MTG). Binary logistic regression analysis predicted poor visual recovery, including decreased GMV in the bilateral insula (right insula: odds ratio [OR] = 17.46, p<0.001; left insula: OR=10.538, p=0.001; respectively) and STG (OR=16.551, p<0.001) and increased ALFF (OR=17.148, p<0.001) and regional homogeneity (OR=10.068, p=0.002) in the left MTG. CONCLUSION: CION patients showed decreased GMV and increased functional activities predominantly located in visual- and cognition-related areas. Decreased GMV and increased ALFF or regional homogeneity in the high-order visual regions (insula, STG, and MTG) are promising imaging markers predicting poor visual outcomes at the 3-year follow-up.

[Comparative analysis of clinical diagnosis application of two intrauterine adhesion scoring criteria].

Objective: To explore the similarities and differences of China Society of Gynecology Endoscopy (CSGE) and American Fertility Society (AFS) intrauterine adhesion (IUA) scoring criteria on IUA grading and their predictive value of reproductive prognosis. Methods: From January 2016 to January 2019, a total of 1 249 patients were diagnosed with IUA by hysteroscopy at Beijing Obstetrics and Gynecology Hospital. Totally, 378 patients with complete clinical data were enrolled, and the results diagnosed by CSGT and AFS scoring criteria were compared and analyzed.And follow-up for 2 years, the pregnancy rate and live birth rate were statistical analysis. Results: (1) The grade of IUA according to AFS and CSGE scoring criteria was less consistent (κ=0.295, P<0.001). Compared with AFS, the proportion of severe IUA cases diagnosed by CSGE was significantly lower [45.8% (173/378) vs 15.1% (57/378); RR=0.22, 95%CI: 0.15-0.30, P<0.01); the proportions of both mild and moderate IUA cases were significantly higher (RR=4.16, 95%CI: 2.38-7.14; RR=2.38, 95%CI: 1.75-3.23; both P<0.01). (2) The pregnancy rates of mild, moderate and severe IUA diagnosed according to CSGE were 11/13, 64.5% (147/228), 31.8% (7/22), live birth rates were 11/13, 54.8% (125/228) and 22.7% (5/22), respectively; there were statistically significant differences between the groups (all P<0.01). The pregnancy rates of mild, moderate and severe IUA diagnosed based on AFS were 3/3, 66.9% (97/145) and 56.5% (65/115), respectively, with no statistically significant difference between the groups (P>0.05). (3) IUA grades based on both CSGE and AFS criteria were significantly negatively correlated with pregnancy rates and live birth rates (CSGE: r=-0.210, r=-0.226; AFS: r=-0.130, r=-0.147; all P<0.05). Univariate logistic regression analysis showed that CSGE had higher OR for both pregnancy rates and live birth rates compared to AFS (3.889 vs 1.657, 3.983 vs 1.554, respectrvely). Conclusions: Compared with AFS, the IUA grade based on CSGE is better related with reproductive prognosis, suggesting that the CSGE standard might be more objective and comprehensive and has better predictive value for reproductive prognosis, thus avoiding overdiagnosis and overtreatment.

[Changes of serum calcium and bone turnover markers in maintenance hemodialysis patients treated with denosumab].

A total of 20 patients (8 males and 12 females) aged (64.8±13.9) years who underwent regular hemodialysis and had bone loss or osteoporosis in Peking University People's Hospital from July to December 2021 were recruited. Sixty milligrams of denosumab were given subcutaneously. The average serum calcium decreased by 0.31 mmol/L and 40% (8/20) of the patients had hypocalcemia one month after treatment. The markers of bone turnover began to decrease 3 days after treatment, and continued to decrease until 5 months after denosumab medication. Multivariate logistic regression analysis failed to detect any independent risk factor for hypocalcemia (R2=0.516, P=0.021). Therefore, denosumab can significantly inhibit bone turnover in hemodialysis patients, however, patients with denosumab medication are more prone to hypocalcemia.

[The prevalence and influence factors of work-related musculoskeletal disorders of dental nurse].

Objective: To investigate the prevalence and Influence factors of work-related musculoskeletal disorders (WMSDs) among dental nurses in tertiary stomatology hospitals. Methods: From June to July 2020, 1321 dental nurses from 18 tertiary stomatology hospitals in 18 provinces of China were enrolled as research subjects through a phased sampling method. The cross-sectional study was performed with validated instruments including the basic information questionnaire, Effort-Reward Imbalance Questionnaire, Nurse-Physician Collaboration Scale, and Chinese Musculoskeletal Questionnaire, so as to get the information about the social demography information, nursing-physician cooperation, work stress and the incidence of WMSDs. The influenang factors of WMSDs was analyzed. Results: The past prevalence, annual prevalence, and weekly prevalence of WMSDs were 80.8% (1067/1321) , 68.7% and 43.7% (575/1321) . 58.4% (771/1321) of nurses had WMSDs in three or more body parts. Neck (51.4%) , waist (42.2%) and shoulder (41.6%) were the top three body regions in the past year. The absenteeism rate due to musculoskeletal disorders ranged from 3.5% to 9.0%, waist (8.9%) and neck (8.0%) were the top two body regions. Working more than 40 hours per week (OR=1.74, P<0.001) and effect/reward>1 (OR=1.50, P=0.002) were risk factors of WMSDs (OR=1.74, 95% CI: 1.32~2.30; OR=1.50, 95%CI: 1.17~1.92, P<0.05) , while regular physical exercise every week (≥30 min per times) was the protective factor (OR=0.74, 95%CI: 0.56~0.97, P=0.030) . Conclusion: The prevalence rate of WMSDs of dental nurses in China was high, and most of the nurses suffered with many body parts. Thus, we should reduce controllable occupational stresso, ensuring adequate rest and strengthening physical exercise to reduce the incidence of WMSDs.

Adverse events following BNT162b2 mRNA COVID-19 vaccination among healthcare workers: A single-centre experience in Malaysia.

INTRODUCTION: The COVID-19 pandemic is a global health crisis that has resulted in a massive disease burden worldwide. Mass vaccination plays an important role in controlling the spread and severity of COVID-19 infections worldwide. MATERIALS AND METHODS: A cross-sectional study was conducted in Hospital Tuanku Ja'afar Seremban between 1 March 2021 and 4 May 2021 to describe the adverse events (AE) following BNT162b2 (Pfizer-BioNTech) vaccination. Healthcare personnel who received at least one dose of the vaccine were invited to complete an online questionnaire. RESULTS: Of 2282 analysed samples, AE were experienced in up to 64.5% (n=1472) of the study participants. Most AE were encountered after the second dose (56.5%, n=832). Pain at the injection site (41.5%, n=944), fever (35.1%, n=798) and lethargy (34.8%, n=792) were the most commonly reported AE. Severe AEFI were reported in a minority (2.9%, n=68). There were no documented anaphylaxis, vaccine-induced thrombosis, or myocarditis. The proportion of female recipients and recipients with a history of allergy were higher in the AE group compared to the non-AE group. CONCLUSION: Our study reinforces the safety of the BNT162b2 mRNA vaccine in the local population. The main adverse events were mild, although they occurred in most patients.

[Observation of microstructure and vessel density changes in the superficial retinal layer in buried optic disc drusen patients].

Objective: To investigate the changes of the microstructure and vascular density in the superficial retinal layer of buried optic disc drusen (ODD) patients. Methods: Retrospective case-control study. A total of 36 ODD eyes (20 patients) and 26 normal control eyes were recruited in Beijing Union Medical College Hospital from January 2018 to July 2020. Measurement of best corrected visual acuity (BCVA), intraocular pressure, slit lamp, fundus examination and visual field examination were performed. The images and data of spectral domain-optical coherence tomography (SD-OCT) and optical coherence tomography angiography (OCTA) were analyzed and summarized. The differences of nasal retinal nerve fiber layer (RNFL), ganglion cell complex (GCC) thickness and macular superficial vascular density (VD) between ODD patients and normal controls were compared by independent sample t-test or Mann Whitney U test (the right eye was selected in bilateral ODD patients). Results: The 20 ODD patients and 26 normal controls were all female. There was no significant difference in age between the two groups (P>0.05). The BCVA and visual field examination was normal in all ODD patients. The SD-OCT examination showed an oval low signal shadow under the nasal outer nuclear layer of the optic disc, or local accumulation like a medium signal shadow with a clear boundary, and a high signal capsule in ODD patients. The RNFL in the upper nasal side of the ODD group was significantly different from the normal control group [(102.6±19.1) µm vs. (119.0±13.8) µm; t=-2.81; P<0.01]. Compared with normal control group [101.0 (100.0, 102.0) µm], the average GCC thickness in the ODD group [97.0 (89.3, 99.8) µm] was significantly different (U=48.50; P<0.01). The OCTA en-face scan showed that the vascular network in the macular area of the affected eyes was sparser than that of the control eyes. There was significant difference in superficial macular VD beteeen the ODD group (48.5%±2.8%) and the control group (51.0%±2.3%) (t=-2.63; P<0.05). Conclusions: There is thinning in the RNFL upper nasal side and GCC layer of the macular region in buried ODD patients, and the superficial VD of the macular region in buried ODD patients is lower than that in the normal controls.

Acceleration of Brain Susceptibility-Weighted Imaging with Compressed Sensitivity Encoding: A Prospective Multicenter Study.

BACKGROUND AND PURPOSE: While three-dimensional susceptibility-weighted imaging has been widely suggested for intracranial vessel imaging, hemorrhage detection, and other neuro-diseases, its relatively long scan time has necessitated the clinical verification of recent progresses of fast imaging techniques. Our aim was to evaluate the effectiveness of brain SWI accelerated by compressed sensitivity encoding to identify the optimal acceleration factors for clinical practice. MATERIALS AND METHODS: Ninety-nine subjects, prospectively enrolled from 5 centers, underwent 8 brain SWI sequences: 5 different folds of compressed sensitivity encoding acceleration (CS2, CS4, CS6, CS8, and CS10), 2 different folds of sensitivity encoding acceleration (SF2 and SF4), and 1 without acceleration. Images were assessed quantitatively on both the SNR of the red nucleus and its contrast ratio to the CSF and, subjectively, with scoring on overall image quality; visibility of the substantia nigra-red nucleus, basilar artery, and internal cerebral vein; and diagnostic confidence of the cerebral microbleeds and other intracranial diseases. RESULTS: Compressed sensitivity encoding showed a promising ability to reduce the acquisition time (from 202 to 41 seconds) of SWI while increasing the acceleration factor from 2 to 10, though at the cost of decreasing the SNR, contrast ratio, and the scores of visual assessments. The visibility of the substantia nigra-red nucleus and internal cerebral vein became unacceptable in CS6 to CS10. The basilar artery was well-distinguished, and diseases including cerebral microbleeds, cavernous angiomas, intracranial gliomas, venous malformations, and subacute hemorrhage were well-diagnosed in all compressed sensitivity encoding sequences. CONCLUSIONS: Compressed sensitivity encoding factor 4 is recommended in routine practice. Compressed sensitivity encoding factor 10 is potentially a fast surrogate for distinguishing the basilar artery and detecting susceptibility-related abnormalities (eg, cerebral microbleeds, cavernous angiomas, gliomas, and venous malformation) at the sacrifice of visualization of the substantia nigra-red nucleus and internal cerebral vein.

[Effects of (+)-catechin and epigallocatechin gallate on alcoholic fatty liver in mice models].

Objective: To investigate the effects of (+)-catechin and epigallocatechin gallate (EGCG) on alcoholic fatty liver in mice models. Methods: Alcohol and high-fat diet were used to construct the model of alcoholic fatty liver disease, and treated the model with (+)-catechin and EGCG. The effects of (+)-catechin and EGCG were analyzed by detecting the contents of TG, TC, MDA, SOD, ALT and AST in serum and pathological changes of liver. Results: (+)-catechin and EGCG could effectively reduce the contents of TG, TC, MDA, ALT and AST, increase the content of SOD in serum of mice with alcoholic fatty liver disease, improve the degree of liver cell edema and reduce the formation of lipid droplets. Conclusion: (+)-catechin and EGCG could repair liver injury and regulate lipid metabolism of hepatocytes in alcoholic fatty liver disease mice models.

[An optimized protocol of meniscus cell extraction for single-cell RNA sequencing].

OBJECTIVE: To optimize the protocol of meniscus cell extraction to enhance the efficiency of cell suspension preparation and maintain a high cell viability for single-cell RNA sequencing. METHODS: We compared the efficiency of the routine cell extraction methods (short-time digestion and long-time digestion) and the optimized protocol for obtaining meniscus cell suspensions by evaluating the cell number obtained and the cell viability. Single-cell RNA sequencing datasets were analyzed to evaluate the stability of the cell suspension prepared using the optimized protocol. The reliability of the optimized protocol was assessed by comparing the single-cell RNA sequencing dataset obtained by the optimized protocol with published single-cell RNA sequencing datasets of the meniscus. RESULTS: The optimized protocol harvested a greater number of cells (over 1×105) than the routine protocols. The cell suspension prepared with the optimized protocol showed a cell viability higher than 80%, the highest among the 3 methods. Analysis of single-cell RNA sequencing datasets showed that the ratio of the mitochondrial genes was below 20% in over 80% of the cells. CD34+ cells, MCAM+ cells and COL1A1+ cells were identified in the datasets. Comparison with the publish datasets showed that the optimized protocol was capable of harvesting COL3A1+, COL1A1+, MYLK+, BMP2+, CD93+ and CDK1+ cells. CONCLUSION: Single-cell suspension prepared from the meniscus can be stably obtained using the optimized protocol for single-cell RNA sequencing using the 10× Genomics platform.

[GPCR48 promotes invasion and metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma cells].

Objective: To observe the G protein-coupled receptor 48 (GPCR48) expression in hepatocellular carcinoma (HCC) cell lines with different metastatic potential and its characteristics effect on the invasion and metastasis of Huh7 hepatoma cells via epithelial-mesenchymal transition (EMT). Methods: Western blot was used to detect the protein expression level of GPCR48 in HCC cells with different metastatic potential. The lentivirus vector expressing GPCR48 gene was constructed. GPCR48 was overexpressed in Huh7 hepatoma cells. The GPCR48 overexpression level was detected by real-time PCR and Western blot. Transwell invasion and migration assay was used to detect the Huh7 hepatoma cells invasion and migration ability in the Control, Mock and GPCR48 overexpression group. Real-time PCR and Western blot were used to detect Huh7 hepatoma cells mRNA and protein expression levels of the EMT related markers (E-cadherin, N-cadherin, vimentin, and γ catenin) in the Control, Mock and GPCR48 overexpression groups, respectively. Analysis of variance was used to compare the differences between data sets. Results: GPCR48 protein expression level in metastatic HCC cell lines was significantly higher than non-metastatic HCC cell lines (P < 0.05). The lentivirus vector expressing the GPCR48 gene had effectively transfected the Huh7 hepatoma cells and stably expressed the GPCR48mRNA and protein. Compared with the Mock and the Control group, Huh7 hepatoma cells invasion and migration ability in the GPCR48 overexpression group was significantly enhanced (F≥5.54, P < 0.05), and the mRNA and protein expression levels of epithelial phenotypic markers E-cadherin and γ-catenin were decreased (P < 0.05). The mRNA and protein expression levels of the mesenchymal phenotypic markers N-cadherin and Vimentin were increased (P < 0.05), indicating that EMT changes occurred in Huh7 hepatoma cells had overexpressed GPCR48. Conclusion: GPCR48 expression level is positively correlated with the metastatic potential of HCC cells. GPCR48 overexpression can down-regulate the expression of epithelial phenotypic markers and up-regulate the expression of mesenchymal phenotypic markers, and induce EMT changes in HCC cells, thus promoting HCC cells invasion and migration.

Study on a new manner of the magnetization switching actuated by a unidirectional pulse current.

A new writing scheme with a unidirectional pulse current is proposed for spin transfer torque (STT) based magnetic random-access memory (MRAM). To investigate the feasibility of the writing scheme, bilayered nano-pillars composed of a soft layer with small in-plane shape anisotropy and a hard layer with either large perpendicular anisotropy (PMA) or in-plane anisotropy (IMA) are designed and their switching behaviors are studied. It is found that in either type of bilayered nano-pillars, with the aid of the attached hard layer, the magnetization of the soft layer can be switched back and forth under a unidirectional pulse current. In an IMA/IMA nano-pillar, the magnetization of the free layer (FL) can achieve excellent alignment, which is in contrast to the IMA/PMA nano-pillar. By optimizing the dimensions and magnetic parameters of the IMA/IMA nano-pillar, a decently low switching current density (4.3 × 1011A m-2) and ultrashort switching time (<1 ns) can be reached. Based on these results, the unidirectional writing scheme is practical if an IMA/IMA bilayer is used to replace the FL in a magnetic tunnel junction. Considering that a unidirectional writing scheme can enable the application of materials with high spin polarization such as half metals, and avoid the injection of writing current into junction using a special design, it may be very promising for STT-MRAM.

[Diagnostic issues of Whipple's disease during chronic inflammatory rheumatism: About three cases]. / Difficultés diagnostiques de la maladie de Whipple au cours des rhumatismes inflammatoires chroniques : trois observations.

INTRODUCTION: Whipple's disease (WD) can mimic chronic inflammatory rheumatism leading to incorrect prescription of tumor necrosis factor inhibitors (TNFI). Several complicated cases of WD have been reported during TNFI treatment which is strongly suspected to modify the host-pathogen relationship. Tropheryma whipplei asymptomatic carriage is high in the general population, making the diagnosis of WD more difficult face to unexplained arthritis. OBSERVATIONS: We report three observations that illustrate situations for which the detection of T. whipplei might be valuable to investigate the differential diagnosis of inflammatory rheumatism. CONCLUSION: The decision to check for T. whipplei infection should rely on individual clinical assessment. It should be considered in the absence of clinical response or in case of worsening of an inflammatory rheumatism under TNFI treatment, especially in front of atypical features. A systematic screening for T. whipplei before anti-TNF treatment seems unjustified since asymptomatic carriers are frequent.

[Inhibiting autophagy by silencing ATG5 and ATG7 enhances inhibitory effect of DDP on DDP-resistant I-10 testicular cancer cells].

OBJECTIVE: To observe the changes in autophagy of cisplatin-resistant I-10 testicular cancer cells (I-10/DDP cells) in response to cisplatin treatment and the effect of silencing ATG5 and ATG7 on autophagy and proliferation of cisplatin-treated cells. OBJECTIVE: I-10/DDP cells treated with 15 µmol/L cisplatin for 12 h were examined for expressions of LC3 and p62 by Western blotting and for autophagy level through transmission electron microscopy and mCherry-GFP-LC3B. I-10/DDP cells were transfected with short hairpin RNAs shRNA-ATG5 or shRNA-ATG7 via Lipfectamine2000, the empty vector (NC group), or Lipfectamine2000 alone (blank control group), and the cellular expressions of ATG5 and ATG7 were detected with Western blotting. The transfected cells were treated with 15 µmol/L cisplatin for 12 h, after that the expressions of LC3 and p62 were detected with Western blotting. Transmission electron microscopy and mCherry-GFP-LC3B were used to detect autophagy level in the cells. MTT assay and colony-forming assay were performed to assess the cell survival fraction and colony formation ability of the treated cells, respectively. OBJECTIVE: After cisplatin treatmert, the expression level of LC3 II increased significantly (P < 0.001), the expression level of p62 decreased (P < 0.05), and the number of autophagosomes increased in I-10/DDP cells. The cells transfected with shRNA-ATG5 or shRNA-ATG7 showed significantly decreased expressions of ATG5 or ATG7 (P=0.005 or P < 0.001). Cisplatin treatment of the transfected cells obviously reduced the cellular expression of LC3 II (P < 0.001), increased the expression of p62 (P < 0.001), and decreased the number of autophagosomes, cell survival fraction and colony formation ability of the cells (P < 0.001). OBJECTIVE: Silencing ATG5 and ATG7 inhibits cisplatin-mediated autophagy and enhances the inhibitory effect of cisplatin on inhibiting cell proliferation.