Intestinal Translocation of Live Porphyromonas gingivalis Drives Insulin Resistance.

Periodontitis has been emphasized as a risk factor of insulin resistance-related systemic diseases. Accumulating evidence has suggested a possible "oral-gut axis" linking oral infection and extraoral diseases, but it remains unclear whether periodontal pathogens can survive the barriers of the digestive tract and how they play their pathogenic roles. The present study established a periodontitis mouse model through oral ligature plus Porphyromonas gingivalis inoculation and demonstrated that periodontitis aggravated diet-induced obesity and insulin resistance, while also causing P. gingivalis enrichment in the intestine. Metabolic labeling strategy validated that P. gingivalis could translocate to the gastrointestinal tract in a viable state. Oral administration of living P. gingivalis elicited insulin resistance, while administration of pasteurized P. gingivalis had no such effect. Combination analysis of metagenome sequencing and nontargeted metabolomics suggested that the tryptophan metabolism pathway, specifically indole and its derivatives, was involved in the pathogenesis of insulin resistance caused by oral administration of living P. gingivalis. Moreover, liquid chromatography-high-resolution mass spectrometry analysis confirmed that the aryl hydrocarbon receptor (AhR) ligands, mainly indole acetic acid, tryptamine, and indole-3-aldehyde, were reduced in diet-induced obese mice with periodontitis, leading to inactivation of AhR signaling. Supplementation with Ficz (6-formylindolo (3,2-b) carbazole), an AhR agonist, alleviated periodontitis-associated insulin resistance, in which the restoration of gut barrier function might play an important role. Collectively, these findings reveal that the oral-gut translocation of viable P. gingivalis works as a fuel linking periodontitis and insulin resistance, in which reduction of AhR ligands and inactivation of AhR signaling are involved. This study provides novel insight into the role of the oral-gut axis in the pathogenesis of periodontitis-associated comorbidities.

Meta-analysis of the effect of whole-body vibration training on the improvement of limb function in patients with Parkinson's disease.

OBJECTIVE: This study aimed to systematically evaluate the effect of whole-body vibration (WBV) training on the improvement of limb function in patients with Parkinson's disease (PD). MATERIALS AND METHODS: An electronic search was performed on the databases, including PubMed, Web of Science, Scopus, SCI-hub, ScienceDirect, Embase, IEEE, Medline, Wiley, ClinicaIKey, CNKI, Wanfang, VIP database, Chinese Medical Association, and CBM Database from inception to May 2022 to collect randomized controlled studies on whole-body vibration training for patients with Parkinson's disease. Two researchers independently screened the literature, extracted the data, and evaluated the quality of the literature, then used ReviewManager 5.4 software for quantitative statistical analysis, including heterogeneity test, sensitivity analysis, risk of bias assessment, combined outcome index effect size and effect size inspection. RESULTS: A total of 9 studies were included in the meta-analysis, including 346 patients, 174 patients in the control group and 172 patients in the observation group. Meta-analysis results showed that, compared with conventional physical therapy or drug therapy alone, whole-body vibration reduced the Unified Parkinson's Disease Rating Scale (UPDRSIII) score and significantly improved the motor function of patients with Parkinson's disease [MD=-2.39, 95% CI (-4.41, -0.37), Z=2.14 (p=0.23)]. Moreover, whole-body vibration significantly improved the walking stability of Parkinson's patients [MD=-1.96, 95% CI (-2.71-1.21), Z=1.17 (p=0.03)]. However, its improvement in balance ability [MD=-0.06, 95% CI (-0.77, 0.65), Z=1.07 (p=0.19)] and daily living ability [MD=0.03, 95% CI (-1.68, 0.74), Z=0.24 (p=0.87)] of patients, it was not t statistically significant. CONCLUSIONS: Compared with conventional therapy, WBV has certain advantages in improving the balance function and gait performance of PD patients, but the effect on balance ability and daily living ability is not significant. Thus, more high-quality research is required for further verification.

First-in-human, phase I study of AK109, an anti-VEGFR2 antibody in patients with advanced or metastatic solid tumors.

BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) plays a key role in antiangiogenesis which has been an essential strategy for cancer treatment. We report the first-in-human study of AK109, a novel anti-VEGFR2 monoclonal antibody, to characterize the safety profile and pharmacokinetics/pharmacodynamics (PK/PD) properties, and explore the preliminary antitumor efficacy in patients with solid tumors. PATIENTS AND METHODS: This was a multicenter, open-label, phase I study, including dose escalation and dose expansion (NCT04547205). Patients with advanced cancers were treated 2 and 3 weekly with escalating doses of AK109. A 3 + 3 design was used to determine the maximum tolerated dose. Blood was sampled for PK/PD analysis. The primary endpoint was safety and recommended phase II dose (RP2D). RESULTS: A total of 40 patients were enrolled. No dose-limiting toxicity was observed. However, 38 patients reported treatment-related adverse events (TRAEs); grade ≥3 TRAEs occurred in 10 patients. The most common TRAEs were proteinuria (n = 24, 60%), hypertension (n = 13, 32.5%), increased aspartate transaminase (n = 11, 27.5%), thrombopenia (n = 10, 25%), and anemia (n = 10, 25%). A total of 28 patients (70%) reported adverse events of special interest (AESIs). The most common AESIs were proteinuria (60%), hypertension (32.5%), and hemorrhage (32.5%), mainly including gum bleeding and urethrorrhagia. AK109 exhibited an approximately linear PK exposure with dose escalation at 2-12 mg/kg. PD analyses showed rapid target engagement. Among the 40 patients, 4 achieved partial response and 21 achieved stable disease with an objective response rate of 10% and a disease control rate of 62.5%. Based on the safety profile, the PK/PD profile, and preliminary antitumor activities, 12 mg/kg Q2W and 15 mg/kg Q3W were selected as RP2D. CONCLUSIONS: AK109 showed manageable safety profile and promising antitumor activity, supporting further clinical development in a large population.

Long noncoding RNA ITGB1 promotes migration and invasion of clear cell renal cell carcinoma by downregulating Mcl-1.

The article "Long noncoding RNA ITGB1 promotes migration and invasion of clear cell renal cell carcinoma by downregulating Mcl-1", by X.-L. Zheng, Y.-Y. Zhang, W.-G. Lv, published in Eur Rev Med Pharmacol Sci 2019; 23 (5): 1996-2002-DOI: 10.26355/eurrev_201903_17238-PMID: 30915742 has been retracted by the author for the following reasons: After the publication of this article, the authors reviewed the process of the experiment and found there were mistakes in the study setting. Authors state that the cancer tissues of 60 inpatients collected in the article included cancer tissues and adjacent tissues. However, the registration and storage of the experiment were not careful, and the cancer tissues were confused with the adjacent tissues. For this reason, the results of this article are not accurate and complete. After consultation among the authors, in line with the rigorous attitude towards scientific research, authors agreed that it was necessary to withdraw the article and make further research and improvement. *After publication, the article was also questioned on PubPeer. Concerns were raised about Figures and in particular Figure 3 which shows overlapping images. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17238.

High prevalence of vertebral deformity in tumor-induced osteomalacia associated with impaired bone microstructure.

PURPOSE: Patients with tumor-induced osteomalacia (TIO) often suffer from irreversible height loss due to vertebral deformity. However, the prevalence of vertebral deformity in TIO patients varies among limited studies. In addition, the distribution and type of vertebral deformity, as well as its risk factors, remain unknown. This study aimed to identify the prevalence, distribution, type and risk factors for vertebral deformity in a large cohort of TIO patients. METHODS: A total of 164 TIO patients were enrolled in this retrospective study. Deformity in vertebrae T4-L4 by lateral thoracolumbar spine radiographs was evaluated according to the semiquantitative method of Genant. Bone microstructure was evaluated by trabecular bone score (TBS) and high-resolution peripheral QCT (HR-pQCT). RESULTS: Ninety-nine (99/164, 60.4%) patients had 517 deformed vertebrae with a bimodal pattern of distribution (T7-9 and T11-L1), and biconcave deformity was the most common type (267/517, 51.6%). Compared with patients without vertebral deformity, those with vertebral deformity had a higher male/female ratio, longer disease duration, more height loss, lower serum phosphate, higher bone turnover markers, lower TBS, lower areal bone mineral density (aBMD), lower peripheral volumetric BMD (vBMD) and worse microstructure. Lower trabecular vBMD and worse trabecular microstructure in the peripheral bone and lower spine TBS were associated with an increased risk of vertebral deformity independently of aBMD. After adjusting for the number of deformed vertebrae, we found little difference in clinical indexes among the patients with different types of vertebral deformity. However, we found significant correlations of clinical indexes with the number of deformed vertebrae and the spinal deformity index. CONCLUSION: We reported a high prevalence of vertebral deformity in the largest cohort of TIO patients and described the vertebral deformity in detail for the first time. Risk factors for vertebral deformity included male sex, long disease duration, height loss, abnormal biochemical indexes and bone impairment. Clinical manifestation, biochemical indexes and bone impairment were correlated with the number of deformed vertebrae and degree of deformity, but not the type of deformity.

Noninvasive prenatal diagnosis for pregnancies at risk for ß-thalassaemia: a retrospective study.

OBJECTIVE: To evaluate the clinical feasibility of noninvasive prenatal diagnosis (NIPD) for ß-thalassaemia using circulating single molecule amplification and re-sequencing technology (cSMART). DESIGN: Through carrier screening, 102 pregnant Chinese couples carrying pathogenic HBB gene variants were recruited to the study. Pregnancies were managed using traditional invasive prenatal diagnosis (IPD). Retrospectively, we evaluated the archived pregnancy plasma DNA by NIPD to evaluate the performance of our cSMART assay for fetal genotyping. SETTING: Chinese prenatal diagnostic centres specialising in thalassaemia testing. POPULATION: Chinese carrier couples at high genetic risk for ß-thalassaemia. METHODS: Fetal cell sampling was performed by amniocentesis and HBB genotypes were determined by reverse dot blot. NIPD was performed by a newly designed HBB cSMART assay and fetal genotypes were called by measuring the allelic ratios in the maternal cell-free DNA. MAIN OUTCOME MEASURES: Concordance of HBB fetal genotyping between IPD and NIPD and the sensitivity and specificity of NIPD. RESULTS: Invasive prenatal diagnosis identified 29 affected homozygotes or compound heterozygotes, 54 heterozygotes and 19 normal homozygotes. Compared with IPD results, 99 of 102 fetuses (97%) were correctly genotyped by our NIPD assay. Two of three discordant samples were false positives and the other sample involved an incorrect call of a heterozygote carrier as a homozygote normal. Overall, the sensitivity and specificity of our NIPD assay was 100% (95% CI 88.06-100.00%) and 97.26% (95% CI 90.45-99.67%), respectively. CONCLUSIONS: This study demonstrates that our cSMART-based NIPD assay for ß-thalassaemia has potential clinical utility as an alternative to IPD for pregnant HBB carrier couples. TWEETABLE ABSTRACT: A new noninvasive test for pregnancies at risk for ß-thalassaemia.

Efficacy and safety of endothelin receptor antagonists in type 2 diabetic kidney disease: A systematic review and meta-analysis of randomized controlled trials.

AIM: To analyse the efficacy and safety of endothelin receptor antagonists for people with diabetic kidney disease. METHODS: Randomized controlled trials comparing endothelin receptor antagonists with placebo in people with diabetic kidney disease were identified through PubMed, Embase and the Cochrane Library. We used a random-effect model to calculate the mean difference or risk ratio with the 95% CI. RESULTS: Seven studies with a total of 4730 participants were included. Overall, endothelin receptor antagonists significantly reduced albuminuria compared with placebo (standardized mean difference -0.48, 95% CI -0.64 to -0.33). Atrasentan, in particular, effectively reduced albuminuria (standardized mean difference -0.58, 95% CI -1.00 to -0.17) and the risk of composite renal endpoints (risk ratio 0.65; 95% CI 0.49 to 0.88), with insignificant change in the rate of congestive heart failure (risk ratio 1.40, 95% CI 0.76 to 2.56) and mortality (risk ratio 1.11, 95% CI 0.77 to 1.61). In contrast, although avosentan reduced albuminuria (standardized mean difference -0.47, 95% CI -0.57 to -0.36) and the risk of composite renal endpoints (risk ratio 0.63, 95% CI 0.42 to 0.94), it was associated with a significant increase in congestive heart failure risk (risk ratio 2.61, 95% CI 1.36 to 5.00) and an insignificant increase in mortality risk (risk ratio 1.50, 95% CI 0.81, 2.78). No significant change in efficacy or safety outcomes with bosentan was detected. Dose-response analysis indicated that 0.75 mg/day atrasentan is expected to be optimal for renoprotection, with maximal albuminuria reduction and minimal fluid retention events. CONCLUSIONS: Among the endothelin receptor antagonists, atrasentan and avosentan, but not bosentan, are effective for renoprotection in people with diabetic kidney disease. Compared with other types and doses, atrasentan 0.75 mg/day is the most promising, with maximal albuminuria reduction and minimal fluid retention. Vigilant monitoring of congestive heart failure risk is needed in future clinical practice. (PROSPERO registration no. CRD42020169840).

Standardization in Quantitative Imaging: A Multicenter Comparison of Radiomic Features from Different Software Packages on Digital Reference Objects and Patient Data Sets.

Radiomic features are being increasingly studied for clinical applications. We aimed to assess the agreement among radiomic features when computed by several groups by using different software packages under very tightly controlled conditions, which included standardized feature definitions and common image data sets. Ten sites (9 from the NCI's Quantitative Imaging Network] positron emission tomography-computed tomography working group plus one site from outside that group) participated in this project. Nine common quantitative imaging features were selected for comparison including features that describe morphology, intensity, shape, and texture. The common image data sets were: three 3D digital reference objects (DROs) and 10 patient image scans from the Lung Image Database Consortium data set using a specific lesion in each scan. Each object (DRO or lesion) was accompanied by an already-defined volume of interest, from which the features were calculated. Feature values for each object (DRO or lesion) were reported. The coefficient of variation (CV), expressed as a percentage, was calculated across software packages for each feature on each object. Thirteen sets of results were obtained for the DROs and patient data sets. Five of the 9 features showed excellent agreement with CV < 1%; 1 feature had moderate agreement (CV < 10%), and 3 features had larger variations (CV ≥ 10%) even after attempts at harmonization of feature calculations. This work highlights the value of feature definition standardization as well as the need to further clarify definitions for some features.

Nasal fluid cytology and cytokine profiles of eosinophilic and non-eosinophilic chronic rhinosinusitis with nasal polyps.

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is a heterogeneous disease with different clinical characteristics and different treatment responsiveness. The aims of this study were to compare the nasal fluid cytology and cytokines between eosinophilic CRSwNP (eCRSwNP) and non-eosinophilic CRSwNP (neCRSwNP) and establish a new multivariate model to predict eCRSwNP before surgery to improve personalized treatment for CRSwNP patients. METHODS: Eighty-six consecutive patients with CRSwNP and sixteen healthy controls were recruited in this study. Nasal fluid (NF) was collected from all subjects and nasal polyp tissue was collected during the surgery. The differential cell counts and concentrations of IL-6, IL-8, TNF-77; and IL-10 in NF were measured. Univariate and multivariate logistic regression were used to identify predictors for eCRSwNP. RESULTS: There were more inflammatory cells in NF of CRSwNP than controls. The eosinophil percentage was significantly higher in eCRSwNP than neCRSwNP and controls. The level of IL-8 was significantly higher in neCRSwNP than in eCRSwNP and controls. Blood eosinophilia, nasal fluid eosinophilia, higher total ethmoid score / total maxillary score (E/M ratio) and higher visual analogue scale (VAS) score of CRS were associated with eCRSwNP, the area under receiver operating characteristic curve (AUC) was 0.800, 0.755, 0.703 and 0.648, respectively. Using the coefficients of multivariate regression, we set up a scoring system to predict eCRSwNP with three of the variates and the AUC was 0.883. CONCLUSION: ECRSwNP, neCRSwNP and healthy controls demonstrated different cytology and cytokine profiles in NF. A new preoperational multivariate prediction model for eCRSwNP with NF eosinophilia, blood eosinophilia and higher E/M ratio was established.

PAX3 silencing suppresses gastric cancer proliferation and angiogenesis via MET/PI3K signaling.

PAX3 is the key factor in cell signal transduction pathway and may be involved in the regulation of cancer cell proliferation, differentiation and migration. The aim of the study was to investigate the effects and mechanism of PAX3 silencing on the gastric cancer. Specific PAX3 silencing was performed both in vitro and in vivo using small-interfering RNAs (siRNAs). The proliferation, apoptosis and angiogenesis of gastric cancer cells were assessed using MTT assay, flow cytometry and in vitro tube formation assay. Mice with gastric xenografts, which expressed either si-PAX3 or non-coding siRNA (si-NC), were developed and the effects of PAX3 silencing on tumor progression were evaluated. PCNA is a proliferating cell nuclear antigen and can be used as an index for evaluating cell proliferation status. Immunocytochemistry assay was used to quantify PAX3 and PCNA expression. After 4 weeks of tumor inoculation, tumor tissues were weighed. Tumor tissue morphology and apoptosis were evaluated using HE staining and TUNEL assay. In order to investigate the effect of silencing PAX3 on cell apoptosis, angiogenesis and MET/PI3K pathway, quantitative real-time PCR (qRT-PCR) or western blot were used to detect the expression levels of caspase-3, VEGF, MET, p-MET, PI3K and p-PI3K. After PAX3 silencing, PAX3 expression was significantly decreased in two gastric cancer cell lines, MKN-28 and SGC-7901 (p<0.05 vs Control). PAX3 silencing reduced cell proliferation, induced cell apoptosis and inhibited tube formation. PAX3 and PCNA expression were also significantly decreased. In mice, silencing PAX3 significantly inhibited tumor growth and decreased microvessel density in tumor. PAX3 silencing also decreased cell density in tumors, which concurred with increased apoptosis and PAX3 expression. PAX3 silencing upregulated the expression of caspase-3, downregulated the expression of VEGF, phosphorylation of PI3K and MET. Our data showed that these anti-tumor effects of PAX3 silencing might be attributed to its role in inducing cell apoptosis and inhibiting angiogenesis.

Focus on autonomic dysfunctions in anti-NMDAR encephalitis: a case report.

OBJECTIVE: We hope it will provide a reference for early detection, early diagnosis, and early treatment of atypical Anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis with non-typical autonomic dysfunctions as the first symptom. PATIENTS AND METHODS: We present a 15-year-old girl with the repetition of conscious disturbance at different levels, but no abnormal movements. Initially, there were no positive findings on routine electroencephalography (EEG) and dynamic video-electroencephalography (V-EEG), but the head-up tilt test (HTT) suggested neurocardiogenic syncope (vascular rejection type), which seemed to be the final diagnosis. However, the patient later experienced several episodes of disturbance of consciousness with unexplained abdominal pain. Abnormalities were discovered on EEG, which indicated the possibility of "epileptic seizures with autonomic-gastrointestinal features". Based on these findings, we finally tested the autoimmune encephalitis-related antibodies for the patient after the literature search and review. RESULTS: The patient was finally diagnosed with anti-NMDAR encephalitis. Her symptoms were fully controlled after glucocorticoid and gamma globulin treatment, and she left the hospital with complete recovery. CONCLUSIONS: Although autonomic nervous dysfunction occurred in our patient, her prognosis was good because she did not have respiratory or (and) circulatory failure. Exclusive diagnosis and early treatment are important in patients with anti-NMDAR encephalitis. Abdominal pain with positive HTT may be a manifestation of autonomic dysfunction in this disease.

Circular circ_0000885 promotes hepatocellular carcinoma proliferation by epigenetically upregulating Caprin1.

OBJECTIVE: Recently, the vital role of circular RNAs is discovered in many diseases, including tumor progression. Hepatocellular carcinoma (HCC) is one of the most ordinary malignant tumors. The purpose of our study is to detect the potential function of circ_0000885 in HCC to offer new biomarkers and targets. PATIENTS AND METHODS: The expression level of circ_0000885 in HCC tissues and cell lines was monitored by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Pearson's Chi-square test was used to determine the association of circ_0000885 expression with several clinicopathological factors. Then knockdown of circ_0000885 was constructed to uncover its function in HCC. The cell growth ability was measured through the cell counting kit-8 (CCK-8) assay, colony formation assay, and cell cycle assay. The Western blot assay was performed to analyze the protein level of Caprin1. RESULTS: Circ_0000885 was highly expressed in HCC tissues than that in adjacent samples. The miR-532-5p expression was associated with lymphatic metastasis and TNM stage. The expression of circ_0000885 was also higher in HCC cell lines. The cell growth ability of HCC cells was inhibited after circ_0000885 was silenced. Furthermore, Caprin1 was inhibited via knockdown of circ_0000885. CONCLUSIONS: Circ_0000885 could enhance cell proliferation and regulate cell cycle of HCC by promoting Caprin1.

LncRNA NEAT1/miR-1224/KLF3 contributes to cell proliferation, apoptosis and invasion in lung cancer.

OBJECTIVE: The aim of this study was to detect the relationship between long-chain non-coding RNA (lncRNA) NEAT1 and microRNA-1224 (miR-1224) in lung cancer and to explore its underlying mechanism. MATERIALS AND METHODS: The expression levels of lncRNA NEAT1 and miR-1224 in lung cancer tissues and cells were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The interaction between lncRNA NEAT1 with miR-1224, miR-1224, and KLF3 was detected by Dual-Luciferase Reporter Gene Assay. MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and flow cytometry were used to detect the changes in the proliferative and apoptosis abilities of lung cancer cells after silencing lncRNA NEAT1 or up-regulating miR-1224, respectively. RESULTS: Compared with adjacent normal tissues, lncRNA NEAT1 was significantly up-regulated, while miR-1224 was significantly down-regulated in lung cancer tissues. LncRNA NEAT1 could specifically bind to the 3'UTR of miR-1224 and regulate its expression. The inhibition of lncRNA NEAT1 remarkably reduced the proliferation and enhanced the apoptosis of lung cancer cells. However, the upregulation of the expression of miR-1224 level could significantly inhibit proliferation and promote the apoptosis rate of lung cancer cells. Furthermore, miR-1224 could downregulate KLF3 expression by directly binding to its 3'UTR. CONCLUSIONS: LncRNA NEAT1 can sponge the expression of miR-1224, thereby affecting the proliferation and apoptosis of lung cancer.

Observation of efficacy of TACE combined with HIFU on patients with middle-advanced liver cancer.

OBJECTIVE: To study the efficacy of transarterial chemoembolization (TACE) combined with high-intensity focused ultrasound (HIFU) in patients with middle-advanced liver cancer. PATIENTS AND METHODS: A total of 100 patients with middle-advanced liver cancer treated in our hospital from January 2015 to January 2018 were selected and randomly divided into TACE group (control group, n=50) and TACE combined with HIFU group (experimental group, n=50) according to different therapeutic regimens. The efficacy was observed after the operation, the blood was collected to detect the postoperative liver function indexes aspartate aminotransferase (AST) and alanine aminotransferase (ALT), the postoperative complications were observed. Also, the immune indexes cluster of differentiation 3+ (CD3+), CD4+, and CD8+ were determined. Moreover, the quality of life (QOL) score was compared between the two groups, the 1-, 2-, 3-, and 5-year survival rates were observed after the operation. Also, the changes in the levels of tumor markers α-L-fucosidase (AFU), alpha-fetoprotein (AFP), carbohydrate antigen 19-9 (CA19-9), and carcinoembryonic antigen (CEA) were observed. RESULTS: In experimental group, the levels of AST, ALT, and blood urea nitrogen (BUN) after the operation were significantly decreased (p<0.05), while the postoperative efficacy was significantly superior to that in control group (p<0.05). The incidence of postoperative complications was significantly reduced (p<0.05), the levels of CD3+, CD4+, CD8+, and natural killer (NK) cells were markedly increased (p<0.05). Also, the QOL score was evidently better than that in control group (p<0.05) and the 1-, 2-, 3-, and 5- year survival rates after the operation were evidently higher than those in control group (p<0.05). After treatment, the levels of AFU, AFP, CA19-9, and CEA were remarkably lower than those before treatment in both groups, while they were remarkably lower in experimental group than those in control group (p<0.05). CONCLUSIONS: TACE combined with HIFU in the treatment of patients with middle-advanced liver cancer can restore the hepatic metabolism, enhance the immunity, improve the QOL, prolong the survival time of patients, and significantly reduce the tumor markers. Also, it has fewer adverse reactions and definite overall efficacy, which is worthy of popularization and application.

Noninvasive fetal genotyping in pregnancies at risk for PKU using a comprehensive quantitative cSMART assay for PAH gene mutations: a clinical feasibility study.

OBJECTIVE: To assess the diagnostic performance of a novel circulating single molecule amplification and re-sequencing technology (cSMART) method for noninvasive prenatal testing (NIPT) of Phenylketonuria (PKU). DESIGN: Blinded NIPT analysis of pregnancies at high risk for PKU. SETTING: Shanghai Xinhua Hospital and Hunan Jiahui Genetics Hospital, China. POPULATION: Couples (n = 33) with a child diagnosed with PKU. METHODS: Trio testing for pathogenic PAH mutations was performed by Sanger sequencing. In second pregnancies, invasive prenatal diagnosis (IPD) was used to determine fetal genotypes. NIPT was performed using a PAH gene-specific cSMART assay. Based on the plasma DNA mutation ratio relative to the fetal DNA fraction, fetal genotypes were assigned using a maximum-likelihood algorithm. MAIN OUTCOME MEASURES: Concordance of fetal genotyping results between IPD and NIPT, and the sensitivity and specificity of the NIPT assay. RESULTS: Compared with gold standard IPD results, 32 of 33 fetuses (96.97%) were accurately genotyped by NIPT. The sensitivity and specificity of the NIPT assay was 100.00% (95% CI 59.04-100.00%) and 96.15% (95% CI 80.36-99.90%), respectively. CONCLUSIONS: The novel cSMART assay demonstrated high accuracy for correctly calling fetal genotypes. We propose that this test has useful clinical utility for the rapid screening of high-risk and low-risk pregnancies with a known history of PKU on one or both sides of the family. TWEETABLE ABSTRACT: NIPT of couples at high risk for PKU using a full-coverage cSMART PAH gene test.

[Effects of sulfur dioxide on alveolar macrophage apoptosis in acute lung injury induced by limb ischemia/reperfusion in rats].

OBJECTIVE: To investigate the effect of sulfur dioxide (SO2) on the apoptosis of alveolar macrophage (AM) in lung protection of limb ischemia/reperfusion (I/R) induced acute lung injury (ALI), and to find a new target for the control of inflammatory response. METHODS: Twenty pathogen-free, adult male Sprague-Dawley (SD) rats (180-230 g) were used in this study. Five rats were to be used for limb ischemia/reperfusion, then plasma was extracted as ischemia/reperfusion serum stimulation. Fifteen rats were to be used for extracting AM by bronchoalveolar lavage. The AM was isolated and cultured, then the cell count was adjusted to 1×106/mL, and randomly divided into the following 4 groups (n=6): control group, I/R group, SO2 group, and I/R+SO2 group. The I/R group was given ischemia/reperfusion serum (500 µg/L) to stimulate 6 h; the SO2 group was given an SO2 donor, Na2SO3/NaHSO3 [(0.54 mmol/kg) / (0.18 mmol/kg)]; and the I/R+SO2 group was given the same ischemia/reperfusion serum and Na2SO3/NaHSO3 at the same time. The level of mitochondrial membrane potential, the state of mitochondrial permeability transition pore (mPTP), the rate of AM apoptosis, the expression of Bcl-2 and Caspase-3 proteins were detected by flow cytometry, microplate reader and Western blotting. RESULTS: Compared with the control group, in the I/R group, the ratio of red to green fluorescence and the absorbance decreased significantly, the percentage of apoptotic cells increased obviously, the apoptotic rate was 43.81%±2.40%, Caspase-3 protein expression increased, Bcl-2 protein expression decreased. While compared with the I/R group, in the I/R+SO2 group, the ratio of red to green fluorescence and the absorbance increased significantly; the apoptotic rate decreased to 37.01%±1.93%, Caspase-3 protein expression decreased, Bcl-2 protein expression increased. CONCLUSION: Exogenous SO2 has the effect of accelerating AM apoptosis by stimulating mPTP to open and mitochondrial membrane potential to decrease; besides, exogenous SO2 could stimulate AM to secrete more anti-inflammatory cytokines and less inflammatory cytokines. In conclusion, exogenous SO2 can reduce macrophage apoptosis by inhibiting mitochondrial pathways.

Long noncoding RNA ITGB1 promotes migration and invasion of clear cell renal cell carcinoma by downregulating Mcl-1.

OBJECTIVE: Researchers have discovered the important role of long noncoding RNA (lncRNAs) in tumorigenesis recently. In this work, we aimed to explore whether lncRNA linc-ITGB1 affected the development of clear cell renal cell carcinoma (ccRCC), and to elucidate the possible underlying mechanism. PATIENTS AND METHODS: Linc-ITGB1 expression in both ccRCC cells and tissue samples was detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Moreover, the association between linc-ITGB1 expression level and patients' disease-free survival rate was explored. Then, wound healing and transwell assays were conducted. Furthermore, the underlying mechanism was explored through RT-qPCR and Western blot assay. RESULTS: Linc-ITGB1 expression level in ccRCC samples was markedly higher than that of the adjacent ones. The expression of linc-ITGB1 was closely related to the disease-free survival time of ccRCC patients. Moreover, the migration and invasion of ccRCC cells were remarkably enhanced after linc-ITGB1 upregulation in vitro. In addition, the mRNA and protein expression of Mcl-1 were significantly downregulated after linc-ITGB1 overexpression. Furthermore, the expression level of Mcl-1 was negatively correlated with the linc-ITGB1 expression in ccRCC tissues. CONCLUSIONS: Our findings suggested that linc-ITGB1 could enhance ccRCC cell migration and invasion via downregulating Mcl-1. In addition, linc-ITGB1 might be a potential therapeutic target for ccRCC.

Serum sclerostin as a potential novel biomarker for heart valve calcification in patients with chronic kidney disease.

OBJECTIVE: To explore the correlation between change in sclerostin level and heart valve calcification in patients with chronic kidney disease (CKD) in stages 3-5, as well as the possible underlying mechanism, which could provide a clinical reference for the diagnosis and treatment of cardiovascular disease (CVD). PATIENTS AND METHODS: 110 patients were divided into a healthy control group and three groups of patients with CKD stages 3, 4, and 5 according to CKD staging guidelines. Scr, BUN, AKP, TC, TG, HDL, LDL, Ca, Pi, and CRP were measured, and calcium-phosphate product (Ca×Pi) calculated. ELISA was used to measure the sclerostin level, and the estimated glomerular filtration rate (eGFR) was calculated by MDRD. Heart valve calcification was measured by a physician in the Cardiac Department of our hospital. The correlations between sclerostin-level change and heart valve calcification, as well as each index in CKD patients in stages 3-5, were analyzed. RESULTS: Compared with the healthy control group, the serum Ca in CKD stage-3, stage-4, and stage-5 groups (p < 0.05) was reduced, and PTH was increased (p < 0.05). Blood Pi and Ca×Pi in the stage-4 and stage-5 groups were increased (p < 0.05). The serum sclerostin level increased with renal hypofunction in stage-3 CKD patients, and was significantly increased compared with that of the control group, reaching the highest level in the terminal stage (p < 0.01). Pearson correlation analysis indicated that serum sclerostin was negatively correlated with eGFR (r = -0.91, p < 0.001) and blood Ca (r= -0.271, p < 0.001), and positively correlated with SCr (r = 0.608, p < 0.001), blood Pi level (r = 0.295, p < 0.001), PTH (r = 0.334, p < 0.001), and Ca×Pi (r = 0.275, p < 0.001). The rate of heart valve calcification in the CKD patients in stage 5 was relatively high (11/30, 36.67%), and significantly higher than that in healthy controls (1/20, 5%; p < 0.01). Logistic regression analysis of heart valve calcification indicated that sclerostin was a risk factor for heart valve calcification in CKD patients in stages 3-5. CONCLUSIONS: The sclerostin level gradually increased with renal hypofunction in CKD patients in stages 3-5, and the increase in serum sclerostin level in the CKD patients occurred earlier than the change in Pi and Ca×Pi. The risk of heart valve calcification in stage-5 CKD patients was significantly increased. Sclerostin is an independent risk factor for heart valve calcification in CKD patients.

Long-term follow-up and treatment of a patient with severe skeletal open bite using temporary anchorage devices.

This case report presents the treatment and long-term follow-up of a patient with severe skeletal hyperdivergent open bite, Class II malocclusion, and a severely retruded chin. After failure of early treatment using high-pull headgear with a bite block during the early permanent dentition stage due to an unfavorable growth pattern, orthognathic surgery was proposed but rejected by the patient. Then, temporary anchorage devices were used to correct the occlusion and establish an acceptable overbite and overjet. The overall observation time was 8.5 years; the treatment time using fixed appliances was 3 years and 4 months. The achieved tooth position and occlusal relationship remained stable 2.5 years later without recurrence of the open bite.

Long non-coding RNA LINC00675 inhibits tumorigenesis and EMT via repressing Wnt/ß-catenin signaling in esophageal squamous cell carcinoma.

OBJECTIVE: Long noncoding RNA LINC00675 (LINC00675) seems to play an anti-oncogenic role in cancers, though its exact function remains unknown. Up to date, little is known about the role of LINC00675 in esophageal squamous cell carcinoma (ESCC). In this study, we aimed to explore the expression pattern, clinical significance and biological function of LINC00675 in ESCC. PATIENTS AND METHODS: RT-PCR was performed to detect the expression levels of LINC00675 in both ESCC tissue and cell lines. The association of LINC00675 expression with clinicopathological factors and prognosis was statistically analyzed. Cell growth was detected by MTT assay and colony formation assay. Cell apoptosis was evaluated with flow cytometry. Migration and invasion ability of ESCC cells were detected wound healing assay and transwell assays. The expressions of EMT-related proteins and Wnt/ß-catenin-related proteins by Western blot were investigated. RESULTS: LINC00675 expression was significantly downregulated in both ESCC tissues and cell lines. Decreased LINC00675 expression was correlated with histological grade, lymph nodes metastasis and advanced clinical stage. Furthermore, LINC00675 could serve as an independent predictor for overall survival in ESCC. Importantly, in vitro experiments indicated that that forced LINC00675 expression significantly suppressed inhibited ESCC cell proliferation, colony formation, migration, invasion and EMT, and promoted cell apoptosis through suppressing Wnt/ß-catenin signaling pathway. CONCLUSIONS: We suggested that LINC00675 acted as a tumor suppressor in ESCC via regulation of Wnt/ß-catenin signaling pathway and may be a new prognostic biomarker and potential therapeutic target for ESCC intervention.