The gut microbiota prime systemic antiviral immunity via the cGAS-STING-IFN-I axis.

The microbiota are vital for immune homeostasis and provide a competitive barrier to bacterial and fungal pathogens. Here, we investigated how gut commensals modulate systemic immunity and response to viral infection. Antibiotic suppression of the gut microbiota reduced systemic tonic type I interferon (IFN-I) and antiviral priming. The microbiota-driven tonic IFN-I-response was dependent on cGAS-STING but not on TLR signaling or direct host-bacteria interactions. Instead, membrane vesicles (MVs) from extracellular bacteria activated the cGAS-STING-IFN-I axis by delivering bacterial DNA into distal host cells. DNA-containing MVs from the gut microbiota were found in circulation and promoted the clearance of both DNA (herpes simplex virus type 1) and RNA (vesicular stomatitis virus) viruses in a cGAS-dependent manner. In summary, this study establishes an important role for the microbiota in peripheral cGAS-STING activation, which promotes host resistance to systemic viral infections. Moreover, it uncovers an underappreciated risk of antibiotic use during viral infections.

Construction of prediction model for fetal growth restriction during first trimester in an Asian population.

OBJECTIVE: To construct a prediction model for fetal growth restriction (FGR) during the first trimester of pregnancy and evaluate its screening performance. METHODS: This was a prospective cohort study of singleton pregnancies that underwent routine ultrasound screening at 11 to 13 + 6 weeks at the Affiliated Suzhou Hospital of Nanjing Medical University between January 2019 and April 2022. Basic clinical information, ultrasound indicators and serum biomarkers of pregnant women were collected. Fetal weight assessment was based on the fetal growth curve for the Southern Chinese population. FGR was diagnosed according to Delphi consensus criteria. Least absolute shrinkage and selection operator (lasso) regression was used to select variables for inclusion in the model. Discrimination, calibration and clinical effectiveness of the model were evaluated in training and validation cohorts. RESULTS: A total of 1188 pregnant women were included, of whom 108 had FGR. Lasso regression identified seven predictive features, including history of maternal hypertension, maternal smoking or passive smoking, gravidity, uterine artery pulsatility index, ductus venosus pulsatility index and multiples of the median values of placental growth factor and soluble fms-like tyrosine kinase-1. The nomogram prediction model constructed from these seven variables accurately predicted FGR, and the area under the receiver-operating-characteristics curve in the validation cohort was 0.82 (95% CI, 0.74-0.90). The calibration curve and Hosmer-Lemeshow test demonstrated good calibration, and the clinical decision curve and clinical impact curve supported its practical value in a clinical setting. CONCLUSION: The multi-index prediction model for FGR has good predictive value during the first trimester. © 2023 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Pediatric Project ECHO® for Pain: implementation and mixed methods evaluation of a virtual medical education program to support interprofessional pain management in children and youth.

BACKGROUND: Pediatric pain is a complex health challenge requiring a multi-modal management approach. It is critical that healthcare providers (HCPs) have access to ongoing, flexible education and mentorship specific to pediatric pain. However, there are significant gaps in available pain education and a need for more opportunities to support interprofessional training. Project Extension for Community Healthcare Outcomes (Project ECHO®) is a model for delivering online HCP education and cultivating a virtual community of practice. Within the pediatric pain setting, ECHO® has potential to improve local access to specialized pain knowledge, particularly among the physicians, nurses, and allied health providers who primarily manage these cases in community and hospital settings across rural and urban environments. The purpose of this study was three-fold. First, to evaluate the feasibility (participation levels, acceptability) of implementing Project ECHO® in the context of pediatric pain. Second, to measure preliminary program impacts on HCP knowledge, self-efficacy, and clinical practice. Third, to characterize HCP program engagement levels before and after onset of the COVID-19 pandemic. METHODS: A needs assessment was conducted to identify interprofessional education gaps and inform the program curriculum. The no-cost Pediatric ECHO® for Pain program offered TeleECHO sessions (didactic and case-based learning) as well as foundational education. Surveys were distributed at baseline and 6 months to assess outcomes using 7-point Likert scales. Participant engagement was assessed for periods prior to and during the COVID-19 pandemic. Descriptive and inferential statistical analyses were conducted. RESULTS: Eighty-five TeleECHO sessions were hosted, with a mean attendance of 34.1 ± 23.4 HCPs. Acceptability scores at 6 months (n = 33) ranged from 5.0 ± 1.4 to 6.5 ± 0.5. Participants reported statistically significant (p < 0.05) improvements in knowledge (7 out of 7 topics) and self-efficacy (8 out of 9 skills). Most participants reported positive practice impacts, including improved satisfaction with managing children with pain. Exploratory analyses showed a trend of greater engagement from ECHO® learners after onset of the COVID-19 pandemic. CONCLUSIONS: Project ECHO® is a feasible and impactful model for virtual education of interprofessional HCPs in managing pediatric pain.

[Targeting microRNA-125b inhibited the metastasis of Alisertib resistance cells through mediating p53 pathway].

Objective: To clarify the mechanisms involvement in Alisertib-resistant colorectal cells and explore a potential target to overcome Alisertib-resistance. Methods: Drug-resistant colon cancer cell line (named as HCT-8-7T cells) was established and transplanted into immunodeficient mice. The metastasis in vivo were observed. Proliferation and migration of HCT-8-7T cells and their parental cells were assessed by colony formation and Transwell assay, respectively. Glycolytic capacity and glutamine metabolism of cells were analyzed by metabolism assays. The protein and mRNA levels of critical factors which are involved in mediating glycolysis and epithelial-mesenchymal transition (EMT) were examined by western blot and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR), respectively. Results: In comparison with the mice transplanted with HCT-8 cells, which were survival with limited metastatic tumor cells in organs, aggressive metastases were observed in liver, lung, kidney and ovary of HCT-8-7T transplanted mice (P<0.05). The levels of ATP [(0.10±0.01) mmol/L], glycolysis [(81.77±8.21) mpH/min] and the capacity of glycolysis [(55.50±3.48) mpH/min] in HCT-8-7T cells were higher than those of HCT-8 cells [(0.04±0.01) mmol/L, (27.77±2.55) mpH/min and(14.00±1.19) mpH/min, respectively, P<0.05]. Meanwhile, the levels of p53 protein and mRNA in HCT-8-7T cells were potently decreased as compared to that in HCT-8 cells (P<0.05). However, the level of miRNA-125b (2.21±0.12) in HCT-8-7T cells was significantly elevated as compared to that in HCT-8 cells (1.00±0.00, P<0.001). In HCT-8-7T cells, forced-expression of p53 reduced the colon number (162.00±24.00) and the migration [(18.53±5.67)%] as compared with those in cells transfected with control vector [274.70±40.50 and (100.00±29.06)%, P<0.05, respectively]. Similarly, miR-125b mimic decreased the glycolysis [(25.28±9.51) mpH/min] in HCT-8-7T cells as compared with that [(54.38±12.70)mpH/min, P=0.003] in HCT-8-7T cells transfected with control. Meanwhile, in comparison with control transfected HCT-8-7T cells, miR-125b mimic also significantly led to an increase in the levels of p53 and ß-catenin, in parallel with a decrease in the levels of PFK1 and HK1 in HCT-8-7T cells (P<0.05). Conclusions: Silencing of p53 by miR-125b could be one of the mechanisms that contributes to Alisertib resistance. Targeting miR-125b could be a strategy to overcome Alisertib resistance.

Detection of blaKPC and blaNDM genes by duplex PCR with lateral flow dipsticks from sterile body fluid samples.

Duplex polymerase chain reaction with lateral flow dipsticks (duplex PCR-LFD) was developed for the simultaneous detection of beta-lactamase Klebsiella pneumoniae carbapenemase (blaKPC ) and beta-lactamase New Dehli metallo-beta-lactamase (blaNDM ) genes in body fluid samples. This method was validated using well-characterized isolates. The assessment of the specificity of duplex PCR-LFD showed that there was no cross-reactivity with other targets. The detection limit of the duplex PCR-LFD assay was 20 CFU per ml for blaKPC and blaNDM . Among 177 sterile body fluid samples tested by the duplex PCR-LFD assay, 40 were blaKPC -positive and five were blaNDM -positive. The results obtained from 122 corresponding Gram-negative bacteria which were isolated from these clinical samples and tested by duplex PCR-LFD assay showed that there were 37 strains carrying blaKPC genes in 40 blaKPC -positive samples and three strains carrying blaNDM genes in five blaNDM -positive samples. Statistical analysis indicated that there was no significant difference between the direct detection of blaKPC and blaNDM genes in clinical sterile body fluid samples and their corresponding clinical isolates. Therefore, duplex PCR-LFD can be effective for the simultaneous detection of blaKPC and blaNDM in clinical isolates and directly from clinical samples, which may be helpful for the administration of appropriate antimicrobial treatment.

Anisotropic spin-orbit torque generation in epitaxial SrIrO3 by symmetry design.

Spin-orbit coupling (SOC), the interaction between the electron spin and the orbital angular momentum, can unlock rich phenomena at interfaces, in particular interconverting spin and charge currents. Conventional heavy metals have been extensively explored due to their strong SOC of conduction electrons. However, spin-orbit effects in classes of materials such as epitaxial 5d-electron transition-metal complex oxides, which also host strong SOC, remain largely unreported. In addition to strong SOC, these complex oxides can also provide the additional tuning knob of epitaxy to control the electronic structure and the engineering of spin-to-charge conversion by crystalline symmetry. Here, we demonstrate room-temperature generation of spin-orbit torque on a ferromagnet with extremely high efficiency via the spin-Hall effect in epitaxial metastable perovskite SrIrO3 We first predict a large intrinsic spin-Hall conductivity in orthorhombic bulk SrIrO3 arising from the Berry curvature in the electronic band structure. By manipulating the intricate interplay between SOC and crystalline symmetry, we control the spin-Hall torque ratio by engineering the tilt of the corner-sharing oxygen octahedra in perovskite SrIrO3 through epitaxial strain. This allows the presence of an anisotropic spin-Hall effect due to a characteristic structural anisotropy in SrIrO3 with orthorhombic symmetry. Our experimental findings demonstrate the heteroepitaxial symmetry design approach to engineer spin-orbit effects. We therefore anticipate that these epitaxial 5d transition-metal oxide thin films can be an ideal building block for low-power spintronics.

[Effect of dNLR and LIPI scores on the prognosis of elderly patients with non-surgical treatment of non-small cell lung cancer].

Objective: To investigate the effects of derived neutrophil to lymphocyte ratio (dNLR) and lung immune prognostic index (LIPI) score on the overall survival (OS) of non-surgical elderly non-small cell lung cancer (NSCLC) patients. Methods: Clinical and pathological data of NSCLC patients in Hebei General Hospital from January 2014 to June 2018 were collected retrospectively. The dNLR value was calculated based on the results of blood routine before treatment, and the optimal cut-off value of dNLR was obtained by ROC curve. The patients were divided into low dNLR level group and high dNLR level group based on the optimal dNLR cut-off value. The groups were classified as good, intermediate and poor based on the LIPI score consisting of lactate dehydrogenase (LDH) and dNLR tested before treatment. The Kaplan-Meier method and Log rank test were used for survival analysis, and the Cox risk proportional regression model was used for analysis of prognostic influences. Results: The area under the ROC curve for dNLR predicting prognosis in non-surgical elderly NSCLC patients was 0.591 (95% CI: 0.491, 0.692; P=0.093). The optimal cut-off value for dNLR predicting prognosis in elderly NSCLC patients was 2.515, with a sensitivity of 45.5% and a specificity of 81.8%. The gender, BMI, pathological type and degree of tumor differentiation were associated with dNLR levels (P<0.05). The median survival times were 16 and 10 months for patients in the low dNLR level group (dNLR<2.51) and high dNLR level group (dNLR≥2.51), respectively (P<0.001), and 15, 10 and 6 months for patients with good, intermediate and poor LIPI scores, respectively (P<0.001). The age, gender, smoking, pathological type, tumor differentiation, clinical stage, BMI, dNLR level, LDH level and LIPI scores were all associated with patient prognosis (P<0.05), and age≥76 years, tumor differentiation and clinical stage Ⅲ and Ⅳ were independent factors influencing patient prognosis (P<0.05). Conclusion: No matter what treatment measures are taken, dNLR level and LIPI score are related to patients' prognosis, and non-surgical elderly NSCLC patients with high dNLR level and poor LIPI score before treatment have worse prognoses.

[A case-control study of occupational noise exposure induced high-frequency hearing loss and the risk of hypertension].

Objective: To investigate the relationship between high frequency hearing loss caused by occupational noise and the risk of hypertension. Methods: In March 2020, a case-control study was conducted. All noise exposed workers who participated in occupational health examination in Wuxi City in 2019 were selected as the study subjects (95432 cases in total) . The hypertension group was defined as the case group, and the normotensive group was defined as the control group. According to the hearing threshold, they were divided into the non high frequency hearing loss group (<40 dB) and the high frequency hearing loss group (≥ 40 dB) . Univariate statistical method and binary logistic regression were used to evaluate the relationship between high-frequency hearing loss and hypertension risk. Stratified analysis was used to compare the risk of hypertension among workers with high-frequency hearing loss of different ages and length of service. Results: There were significant differences in gender, age, length of service, enterprise scale, economic type and high-frequency hearing loss between control group and hypertension group (P<0.05) . Binary logistic regression analysis showed that after adjusting for gender, age, length of service, enterprise scale and economic type, the risk of hypertension in the high-frequency hearing loss group was still increased (OR=1.062, 95%CI: 1.007~1.121, P=0.027) . The risk of hypertension in high-frequency hearing loss patients was higher than that in non high-frequency hearing loss patients in 20-39 years old and 40-59 years old age groups (OR=1.536, 95%CI: 1.353~1.743; OR=1.179, 95%CI: 1.111~1.250; P<0.05) . The risk of hypertension in high-frequency hearing loss patients in <5years, 5-9years, 10-14 years, 15-19 years and ≥20 years working age groups were higher than that in non high-frequency hearing loss groups (OR=1.926, 95%CI=1.007-1.121; OR=1.635, 95%CI=1.478-1.810; OR=1.312, 95%CI=1.167-1.474; OR=1.445, 95%CI=1.238-1.686; OR=1.235, 95%CI=1.043-1.463; P<0.05) . Conclusion: There is a certain relationship between high-frequency hearing loss caused by occupational noise and the risk of hypertension, and the risk of hypertension is different among high-frequency hearing loss patients of different ages and working years.

[Application of acellular dermal matrix loaded collagen binding domain-vascular endothelial growth factor patch in urethral wound repair in beagles].

Objective: To investigate the application value of acellular dermal matrix (ADM) loaded collagen binding domain-vascular endothelial growth factor (CBD-VEGF) patch in the repair of urethral wound in beagles. Methods: Twenty-one healthy adult beagles were selected to establish urethral wound model and randomly divided into 3 groups, which were treated with different repair methods. CBD-VEGF patch was prepared and instantaneous implantation was performed as the models were successfully built. The blank control group was treated without any repair materials;the ADM group was treated with collagen patch for repair and the ADM loaded with CBD-VEGF group was treated with collagen patch loaded with VEGF for repair. The safety and effectiveness of the three groups of materials implanted into the urethral wound model at 6 months after surgery were compared. Urodynamic examination, urethral angiography and pathological examination were used to evaluate the recovery of urethral tissue in the three groups. Result: Compared with blank control group and ADM patch group, there was no complications or other adverse conditions in the ADM-loaded CBD-VEGF patch group. Urethrography was performed to measure the diameter of the urethra in the experimental segment, the mean urethral diameter of blank control group [(3.07±0.43) mm] was significantly smaller than that of ADM patch group [(3.73±0.11) mm] and ADM loaded CBD-VEGF patch group [(3.64±0.32) mm], with statistical difference (P<0.05). Pathological examination with HE and Masson staining showed more proliferative blood vessels in the ADM loaded CBD-VEGF patch group. HE staining indicated that there were more proliferative vessels in the group of CBD-VEGF patch loaded with ADM. Masson staining showed that the positive area percentage of collagen tissue in the three groups was 34.27%±7.40%, 29.08%±3.79% and 28.02%±2.39%, respectively, with no statistical difference between the three groups(all P>0.05). There was no excessive deposition of collagen tissue in the two groups using collagen patch. The percentage of positive staining per unit area in the three groups were 1.4%±0.1%,1.8%±0.1%3.1%±0.2%, respectively, and the difference was statistically significant among three groups and between every two groups (all P<0.01), indicating the vascular density of CBD-VEGF group loaded with ADM was the highest. Conclusion: ADM loaded CBD-VEGF patch is a good biological support material for covering the wound after urethral surgery, and it provides an alternative direction for promoting the healing of the urethral wound.

[Genomic investigation of human Streptococcus suis infection in Shandong Province].

To investigate Streptococcus suis (S.suis) isolated from patients in Shandong province using genomic epidemiology and pathogenologic analysis. To provide the foundation to establish reasonable and accurate prevention and control measures of human S. suis infection. Molecular typing, whole genome phylogenetic tree, virulence gene typing, antibiotic resistance profile and mobile genetic elements carrying antibiotic resistance genes of isolated S. suis strains were investigated. The pathogenicity of isolated strains was also evaluated by comparing their capacity to induce pro-inflammatory cytokine production in vitro. S. suis infections in Shandong province were predominantly due to serotype 2 and sequence type 1 strains. The major symptoms were meningitis. The studied strains could be divided into five lineages. All strains belong to highly pathogenic type in Shandong province,Strains from lineage 2 possessed higher capacity to stimulate pro-inflammatory cytokine production than other strains did, even though other strains belong to highly pathogenic strains. In addition, multiple antibiotic resistance genes and corresponding mobile genetic elements werewidespread in S. suis strains from Shandong province, except strains from lineage 3. High diversities in genome, evolutionary path and pathogenicity of S. suis strains from Shandong province were revealed. It was necessary to surveillant the S. suis strain in genomic level.

[Survival analysis of silicosis patients in Wuxi City].

Objective: To analyze the living condition and influencing factors of silicosis patients in Wuxi City form 1975 to 2019. Methods: Through the monitoring of death causes of residents, the paper-based materials and online report system of diagnosis over the years, and the combination of public security and human social system to obtain 3721 cases of silicosis patients as subjects form August to December 2019. And the combination of Kaplan Meier method and life table method were used to carry out single factor survival analysis. Through Cox regression model to analyze the factors affecting the survival time of patients. Results: From 1975 to 2019, 3721 cases of silicosis and 1274 deaths have been reported in Wuxi City, with a mortality rate of 34.24% and a median survival time of 30.9 years. With the development of diagnosis time, the mortality decreased significantly (χ(2)=747.75, P<0.05) . Compared with the first stage silicosis patients, the risk of decreased survival time of the third stage silicosis patients increased (HR=1.486, P<0.05) . Compared with the non-smoking patients, the risk of decreased survival time of the smoking patients increased (HR=1.136, P<0.05) . Compared with the patients who were less than 30 years old, the risk of decreased survival time of patients with 40-49 years old, 50-59 years old and more than 60 years old were increased (HR=9.641, 13.650, 26.794, P<0.05) . Compared with the patients who received industrial and commercial insurance, the risk of decreased survival time of patients who received compensation from employers, basic medical insurance for urban and rural residents, other social compensation and no compensation were increased (HR=3.137, 3.119, 5.129, 8.442, P<0.05) . Conclusion: The survival time of silicosis patients is related to the stage of silicosis, smoking condition, age of onset and social compensation. We should focus on controlling the above risk factors so as to improve the quality of life of patients and prolong their lives.

[Comparison of different reconstruction procedures after distal gastrectomy in patients with gastric cancer].

Objective: To systematically compare the effect of Roux-en-Y with Billroth Ⅰ or Billroth Ⅱ in gastric cancer patients after distal gastrectomy by meta-analysis. Methods: Systematic search was conducted on the relevant electronic databases of Pubmed, Embase, Wanfang Database, CNKI and VIP from the established time to August 18, 2019. The randomized controlled trials about comparison of Roux-en-Y with Billroth Ⅰ or Billroth Ⅱ were strictly screened and analyzed by the software of Revman 5.3. Procedure and postoperative outcomes were analyzed, respectively. Results: A total of 783 relevant literatures were systematically retrieved, and 6 randomized controlled trials, including 954 patients, finally met the inclusion criteria after strict screening. The results of meta-analysis showed that operative time of Billroth Ⅰ was significantly shorter than that of Roux-en-Y (MD=-37.60, 95%CI:-50.79--24.40, P<0.001), intraoperative bleeding (MD=-21.64, 95%CI:-32.20--11.07, P<0.001) and the number of delayed gastric emptying (RR=0.52, 95%CI: 0.31-0.86, P=0.01) of Billroth Ⅰ were both significantly less than those of Roux-en-Y, while bile reflux (RR=8.17, 95%CI: 2.21-31.53, P=0.002) and residual gastritis (RR=1.75, 95%CI:1.43-2.14, P<0.000 01) of Billroth Ⅰ were both significantly higher than those of Roux-en-Y, other outcomes showed no significant difference. Compared with Roux-en-Y, operative time of Billroth Ⅱ was significantly shorter (MD=-19.73, 95%CI:-32.82--6.64, P=0.003), while bile reflux (RR=17.63, 95%CI: 4.50-69.02, P<0.001), residual gastritis (RR=1.94, 95%CI:1.15-3.26, P=0.01) and reflux esophagitis (RR=3.13, 95%CI: 1.31-7.45, P=0.01) of Billroth Ⅱ were all significantly higher, and there was no significant difference in other outcomes. Conclusion: Compared with Billroth Ⅰ and Billroth Ⅱ, the operation time of Roux-en-Y in gastric cancer patients undergoing distal gastrectomy is longer, but the incidences of bile reflux and residual gastritis are both lower, and the postoperative quality of life seems better.

[Prognostic value of new TNM staging and serum CA125 in malignant peritoneal mesothelioma].

Objective: To explore the relationship between the new Tumor-Node-Metastasis (TNM) staging system and the serum CA125 level with the prognosis of malignant peritoneal mesothelioma (MPeM) . Methods: The clinical data of 74 patients with MPeM diagnosed by pathology and immunohistochemistry were collected from January 2005 to June 2016 in Cangzhou Central Hospital. According to the results of CT-peritoneal carcinoma index (PCI) , the tumor load was divided into T1 (PCI 1-10) , T2 (PCI 11-20) , T3 (PCI 21-30) and T4 (PCI 31-39) , combined with lymph node metastasis and extraperitoneal metastasis, a new TNM staging system was established. And serum CA125 level was measured in the same time. The median survival time of patients with MPeM, the effect of the new TNM staging system, and serum CA125 levels on their prognosis were retrospectively analyzed. Results: Among the 74 patients with MPeM, 25 (33.8%) cases were males and 49 (66.2%) cases were females. There were 8 cases with systemic chemotherapy, 8 cases with heated intraperitoneal chemotherapy, and 1 case with combination chemotherapy. 10 cases were T1, 22 cases were T2, 27 cases were T3, 15 cases were T4, 12 cases had lymph node metastasis and 10 cases had distant metastasis. The median survival time of T1, T2, T3 and T4 were 12, 10, 6 and 3 months respectively. There were 38 (77.6%) cases with high serum CA125 in all 49 cases who have been tested for CA125. The median survival time of positive group and negative group were 6 months and 11 months respectively. 68 (91.9%) patients had died by the end of collecting data. The median survival time was 8 months. Univariate analysis showed that there were significant differences in survival time between patients with different CT-PCI stages, serum CA125 levels, and with or without lymph node and extraperitoneal metastasis (P<0.05) . Multivariate analysis showed that CT-PCI was independent risk factors for the prognosis of MPeM (HR=2.203, 95%CI: 1.475-3.289) . Conclusion: The new TNM staging system and serum CA125 are important for the prognosis of patients with MPeM. Early detection, early diagnosis and comprehensive treatment can improve the survival time of patients with MPeM.

Clock mutant promotes osteoarthritis by inhibiting the acetylation of NFκB.

OBJECTIVES: To examine the effect of the circadian gene Clock on posttranscriptional function and pro-inflammatory mechanisms in osteoarthritis (OA). METHODS: The cartilage from Clock mutant mice was assessed using histology, (OA) score, and real-time polymerase chain reaction (PCR) quantification of key pro-inflammatory genes. Nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) translocation, posttranslational state and expression levels during day and night conditions were assessed using immunoblot and IP. The regulation of transcription by Clock in cartilage tissue was assessed by using chromatin immunoprecipitation (ChIP) and luciferase assays. Total acetylation level and pattern over 24 h were quantified using immunoblot and real-time PCR. Finally, the effects of exogenous Clock nanoparticle treatment were quantified by histology and immunoblot. RESULTS: The Clock mutation significantly promoted the degradation of cartilage and the expression of the key pro-inflammatory mediators, IL-1ß, IL-6 and MCP-1. The Clock mutation significantly promoted NFκB nuclear translocation. The circadian protein CLOCK positively regulates NFκB at the transcriptional level by binding the E-box domain. The Clock mutation significantly inhibited the total lysine acetylation level in cartilage and inhibited NFκB acetylation at the Lys310 residue but promoted phosphorylation at the Ser276 residue. The forced expression of Clock in vivo inhibited NFκB activation by increasing acetylation and decreasing phosphorylation levels and by decreasing cartilage damage and inflammation. CONCLUSIONS: This study demonstrates the mutation of Clock promotes inflammatory activity by mediating the posttranscriptional regulation of NFκB in OA pathogenesis.

The PNKD gene is associated with Tourette Disorder or Tic disorder in a multiplex family.

Tourette Disorder (TD) is a childhood-onset neuropsychiatric and neurodevelopmental disorder characterized by the presence of both motor and vocal tics. The genetic architecture of TD is believed to be complex and heterogeneous. Nevertheless, DNA sequence variants co-segregating with TD phenotypes within multiplex families have been identified. This report examines whole exomes of affected and unaffected individuals in a multiplex TD family to discover genes involved in the TD etiology. We performed whole exome sequencing on six out of nine members in a three-generation TD multiplex family. Putative deleterious sequence variants co-segregating with TD patients were identified by our in-house bioinformatics pipeline. Induced pluripotent stem cells (iPSCs) were generated from one unaffected and two TD affected individuals. Neurons were derived from the iPSCs and biochemical assays were conducted to evaluate possible molecular differences between affected and unaffected. A rare heterozygous nonsense mutation in PNKD was co-segregated with TD in this multiplex family. Transcript and protein levels of the PNKD long isoform were reduced in neurons derived from the individuals with TD due to the nonsense mutation, indicating nonsense-mediated mRNA decay. We demonstrated that the PNKD long isoform monomer oligomerizes with itself as well as interacts with the synaptic active zone protein RIMS1α. We concluded that reduced PNKD long isoform levels are detected in all affected individuals and we provide evidence for a mechanism whereby this might contribute to the TD phenotype.

[Iodine status of vulnerable populations in Henan Province of China three years after the implementation of new iodized salt standard].

Iodine nutrition surveillance of vulnerable population was conducted in 18 cities of Henan Province in 2015. The medians and quartiles of urinary iodine concentration in women of reproductive-age (n=3 318), pregnant women (n=36 366), lactating women (n=4 153), infants <2 years of age (n=1 017), and children aged 8-10 years (n=4 806) were 202.1(124.0, 310.0), 192.4(133.6, 262.4), 168.3(119.1, 248.3), 214.2(156.1, 272.3) and 90.0 (121.8, 285.6) µg/L, respectively. Iodine status of the vulnerable populations was generally regarded as adequate in Henan Province. But the median of urinary iodine concentration of reproductive-age women were slightly above the adequate level. The proportions above iodine adequate level were 26.7% in reproductive-age women, 29.4% in pregnant women and 22.5% in children aged 8-10 years.

MRI-measured myocardial iron load in patients with severe diabetic heart failure.

AIM: To investigate whether a correlation exists between abnormal myocardial iron status and cardiac lipid deposition as well as other biomarkers in patients with diabetic heart failure (DHF). MATERIALS AND METHODS: Seventeen volunteers (Group 1), 26 patients with non-severe DHF (Group 2), and 25 patients with severe DHF (Group 3) were recruited for this study. Myocardial middle-section T2* mapping and septal 1H magnetic resonance spectroscopy (1H-MRS) were performed using a 3 T magnetic resonance imaging (MRI) machine to assess the iron status and lipid deposition individually. Fasting venous blood was used to examine serum biomarkers. RESULTS: Cardiac T2* (ms) of the three groups were 22.8±2.1, 21.7±1.8, and 18.6±1.3, respectively. The value of Group 3 was significantly lower than that of the other two groups (p<0.001). Myocardial triglyceride (%) levels differed among the three groups (Group 1, 0.53±0.13; Group 2, 1.11±0.29; Group 3, 1.47±0.12; p<0.001). Cardiac T2* was inversely correlated with both cardiac triglycerides and left ventricular ejection fraction (LVEF) in overall participants (Groups 1-3) or Group 3 (each p<0.001). CONCLUSIONS: Abnormal myocardial iron status was found in patients with severe diabetic heart failure. Myocardial lipotoxicity may be responsible for this process.

[Diagnostic value of protein markers in plasma exosomes of lung squamous cell carcinoma].

Objective: To investigate the tumor-associated protein molecules carried by plasma exosomes of patients with lung squamous cell carcinoma before treatment and analyze their value as clinical markers. Methods: Exosomes from 2 patients with lung squamous cell carcinoma before treatment and 2 healthy controls were collected by ultracentrifugation. Proteomics was applied to analyze the protein expression profiles of exosomes. Candidate molecules were verified in another 30 exosomes samples from lung squamous cell carcinoma and healthy controls using enzyme-linked immunosorbent assay (ELISA). Results: Electron microscopy and particle-counting assay showed that high-quality exosomes were collected. The number of exosomes distributed from 45 to 135 nm in 2 cases of lung cancer patients were 7.89×10(11)/ml and 9.71×10(11)/ml, respectively, significantly higher than 2.76×10(11)/ml and 1.41×10(11)/ml in healthy controls. Proteomic analysis showed that proteins of exosomes in lung squamous cell carcinoma patients were very different from those of healthy controls, and some proteins are related to important functions in tumor progression. 14-3-3ζ from exosomes was selected and further verified as a marker, and the area under the receiver operating characteristic curve (ROC) was 0.68. The sensitivity and specificity of 14-3-3 ζ from exosomes were 60.0% and 80.0%, respectively, suggested that it could be used as a diagnostic marker for lung squamous cell carcinoma. Conclusion: The exosome counts in plasma and the protein molecules from exosomes, such as 14-3-3ζ, are closely related to the tumorigenesis, which can be used to assist clinical diagnosis of lung squamous cell carcinoma patients.

[Clinical relevance of common inhibitory immune checkpoint genes in esophageal squamous cell carcinoma].

Objective: To analyze the potential clinical relevance of inhibitory immune checkpoint genes, including PD1, PD-L1, CTLA4, LAG3 and TIM3 in esophageal squamous cell carcinoma (ESCC). Methods: Expression were extracted from mRNA microarray data from 119 paired ESCC and adjacent normal tissues, which were collected from National Cancer Center/Chinese Academy of Medical Sciences between December 2005 and December 2007.Differential expression analyses and survival analyses were then performed to identify potential clinical relevance using R software. Results: Expression of PD-L1, CTLA4, LAG3 and TIM3 were significantly upregulated in ESCC tissues compared with adjacent normal tissues (P<0.05). CTLA4 was significantly correlated with tumor location and age (P<0.05). PD1 and LAG3 were significantly associated with overall survival and were favorable prognostic factors in ESCC. Conclusions: Inhibitory immune checkpoint genes are commonly overexpressed in ESCC and are significantly associated with patients overall survival.These results provide important references in the application of immune checkpoints therapy in ESCC.

[Role and mechanism of lipopolysaccharide induced exosome in the pathogenesis of acute lung injury].

Objective: To explore the effect and mechanism of exosome derived from lipopolysaccharide (LPS)-induced mouse macrophage (RAW264.7) on acute lung injury. Methods: RAW264.7 were cultured in vitro and divided into 2 groups: control group and LPS-induced group. The exosomes were extracted from the two groups of cell supernatant by ultracentrifugation and classified into 2 groups: C-EXO group and LPS-EXO group. In vivo, random allocation was used to averagely divide the eighteen male C57BL/6 mice into 3 groups: control group, EXO-control group and EXO-LPS group. All mice were sacrificed after 12 h. The lung tissue was used for HE staining to assess the degree of acute lung injury as well as immunohistochemical staining for interleukin (IL) -1ß and tumor necrosis factor (TNF)-α. The tissue protein expression levels of IL-1ß, TNF-α, ß-catenin, E-cadhein, ZO-1 and Occludin were measured by Western blot. In vitro, alveolar type Ⅱ epithelial cells (MLE-12) were cultured and divided into 3 groups: C-control group, EXO-control-induced group and EXO-LPS-induced group. The tissue protein expression levels of IL-1ß, TNF-α, and Occludin were measured by Western blot after 12 h. Results: The two samples of C-EXO group and LPS-EXO group was proved to be exosomes. Under a light microscope, the lung tissue of EXO-LPS group showed inflammatory cell infiltration, hemorrhage, interstitial and alveolar edema, and the thickness of alveolar septum. The tissue protein levels of IL-1ß and TNF-α in EXO-LPS group were obviously higher than the control group, EXO-control group (1.331±0.203 and 0.274±0.018, 0.892±0.074; 0.800±0.096 and 0.596±0.025, 0.441±0.061; all P<0.05). While the tissue protein levels of Occludin showed the opposite phenomenon (0.251±0.021 and 0.862±0.029, 0.453±0.013; all P<0.05). In vitro, Compared with the C-control group and the EXO-control-induced group, the expression levels of IL-1ß and TNF-α increased significantly in the EXO-LPS-induced group (0.900±0.033 and 0.320±0.030, 0.661±0.028; 0.739±0.045 and 0.151±0.024, 0.360±0.037; all P<0.05). whereas the protein levels of Occludin expression were reversed in MLE-12 (0.585±0.082 and 0.941±0.090, 0.732±0.083; all P<0.05). Conclusion: Exosomes derived from LPS-induced RAW264.7 can induced the acute lung injury by affecting barrier function, which probably is related to the low degree of Occludin in alveolar type Ⅱ epithelial cells.