Objective To investigate the effect of intra-articular tumor necrosis factor (TNF) inhibitor injection in patients with moderate to severe rheumatoid arthritis (RA) and values of power Doppler ultrasonography in evaluating effect of intra-articular injection.Methods RA patients with arthritis in knee and/or elbow and/or ankle referred to the Department of Rheumatology in the First Affiliated Hospital of Xi'an Jiaotong University were enrolled to receive intra-articular injection with 50 mg or 25 mg of recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein (TNFR:Fc) for injection after synovial fluid aspiration.Evaluation of visual analogue scale for pain of the involved joints,erythrocyte sedimentation rate (ESR),C reactive protein (CRP) and 28-joint disease activity score (DAS28) were performed before and after intra-articular TNFR:Fc injection.Synovial hypertrophy,power Doppler signal and joint effusion were analyzed and graded by ultrasound before and after intra-articular TNFR:Fc injection.Comparisons of continuous data between groups was made by t test.The data that were not normally distributed was analyzed by Mann-Whitney U rank sum test.Results Fifty-four patients with RA [6 men and 48 women,mean age (52±11) years,mean duration of disease (7±3) years] were included in this study.A significant decrease in visual analogue scale for pain of the involved joints (t=2.630,P=0.018;t=2.160,P=0.043),ESR (t=2.094,P=0.030;Z=-2.242,P=0.030),CRP (Z=-2.199,P=0.030;Z=-3.337,P=0.001) and DAS28 (t=3.579,P=0.002;t=5.538,P=0.000) were observed after one month of injection of 50 mg or 25 mg of TNFR:Fc.Synovial hypertrophy (t=2.175,P=0.036;t=2.280,P=0.030) power Doppler signal (t=2.500,P=0.020;Z=-2.504,P=0.013) and joint effusion (Z=-1.790,P=0.042;t=2.230,P=0.027) were reduced significantly after one month of intra-articular TNFR:Fc injection in knee.Synovial hypertrophy (t=2.180,P=0.034;t=2.480,P=0.030) and power Doppler signal (t=2.681,P=0.020;t=5.482,P=0.000) were also reduced significantly after one month of intra-articular TNFR:Fc injection in elbow and ankle.Conclusion Intra-articular TNFR:Fc injection is an effective and safe treatment in RA patients with monoarthritis.Ultrasound may be an objective and valid method in evaluating the effect of intraarticular TNF inhibitor injection in RA patients.

Multi-walled carbon nanotubes-tungsten oxide (MWCNTs-WOx) nanocomposites were fabricated on glassy carbon electrode (GCE) through a simple electrodeposition method,in which WOx were fabricated on MWCNTs. The morphology and constitution were characterized by field emission scanning electron microscopy(SEM) and X-ray photoelectron spectroscopy(XPS). Electrochemical characterization of modified electrode was done by electrochemical impedance spectroscopy (EIS). The cyclic voltammogram (CV)method was adopted to investigate the electrochemical behavior of dopamine (DA) on MWCNTs-WOx-modified glassy carbon electrode, and a new detection method for DA was developed by differential pulse voltammetry (DPV). The results showed that MWCNTs-WOx nanocomposites had obvious electrocatalytic effect on DA in phosphate buffer solution (pH=6.5). Under the optimized experimental conditions, the DA peak current demonstrated a good linear relationship with concentration in the range of 0.05-1.00 mmol/L, and the detection limit was 17 μmol/L(S/N=3). Effects of different experimental parameters on the response current of the modified electrode were investigated,and it was found that the prepared electrochemical sensor displayed good reproducibility,high selectivity and strong anti-interference ability. UA did not interfere with the detection of DA. A new electrochemical method for the quantitative determination of DA was established and successfully applied to the determination of dopamine hydrochloride injection samples.

OBJECTIVE: To explore the genetic etiology of a child with lymphangiectasia and lymphedema. METHODS: DNA sample of the patient was extracted and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing. RESULTS: The patient was found to carry compound heterozygote variants (c.521G>A and c.472C>T) of the CCBE1 gene, which were respectively inherited from his parents. CONCLUSION The compound heterozygote variants of the CCBE1 gene probably underlie the disease in this child.

OBJECTIVE: To explore the genetic basis for a pedigree affected with X-linked mental retardation. METHODS: The proband was subjected to chromosomal karyotyping, FMR1 mutation testing and copy number variation analysis with a single nucleotide polymorphism microarray (SNP array). His family members were subjected to multiplex ligation-dependent probe amplification (MLPA) assaying. Expression of genes within the repeated region were analyzed. RESULTS: The proband had a normal chromosomal karyotype and normal number of CGG repeats within the FMR1 gene. SNP array identified a 370 kb duplication in Xq28 (ChrX: 153 027 633-153 398 515), which encompassed 14 genes including MECP2. The patient was diagnosed as Lubs X-linked mental retardation syndrome (MRXSL). MLPA confirmed the presence of copy number variation, its co-segregation with the disease, in addition with the carrier status of females. Genes from the duplicated region showed higher levels of expression (1.79 to 5.38 folds) within peripheral blood nucleated cells of the proband. CONCLUSION The patients were diagnosed with MRXSL. The expression of affected genes was up-regulated due to the duplication. Genetic counseling and prenatal diagnosis may be provided based on the results.
DNA Copy Number Variations ; Female ; Fragile X Mental Retardation Protein ; Humans ; Mental Retardation, X-Linked ; Methyl-CpG-Binding Protein 2 ; Pedigree ; Pregnancy

OBJECTIVE: To explore the genetic basis for a child with 46,XY disorders of sex development (DSD) and explore its genotype-phenotype correlation. METHODS: The child was subjected to whole exome sequencing (WES), and exons 1 to 7 of NR5A1 were subjected to multiplex ligation-dependent probe amplification (MLPA) analysis. RESULTS: The patient presented with rudimentary vulva of a female with Tanner stage 1. B-mode ultrasonography has detected ovary and uterus. The child was found to have a chromosome karyotype of 46,XY. WES revealed that the patient has harbored heterozygous deletion of exon 5 of the NR5A1 gene, which was a novel pathogenic variant inherited from the mother. No abnormality was found in the father. CONCLUSION The main symptoms of 46,XY DSD children are insufficient external genitalia masculinization, for which variants of the NR5A1 gene are an important cause. WES has improved the detection rate of genetic variants and provided a solid basis for genetic counseling of the affected families.
Child ; Disorder of Sex Development, 46,XY/genetics* ; Disorders of Sex Development/genetics* ; Exons/genetics* ; Female ; Genetic Testing ; Heterozygote ; Humans ; Mutation ; Steroidogenic Factor 1/genetics*

Objective:To invetigate the influencing factors and clinical significance of liver function damage (LFD) in patients diagnosed with Corona Virus Disease 2019 (COVID-19).Methods:The retrospective case-control study was conducted. The clinicopathological data of 51 patients with COVID-19 who were admitted to the Sino-French New City Branch of Tongji Hospital Affiliated to Huazhong University of Science and Technology by the 5th group assisting team from the First Hospital of Jilin University from February 9th to 27th in 2020 were collected. There were 27 males and 24 females, aged from 36 to 86 years, with an average age of 68 years. The treatment modality was according to the diagnostic and therapeutic guideline for COVID-19 (Trial 6th edition) issued by National Health Commission. Observation indicators: (1) clinical data of patients; (2) analysis of liver function index and treatment of LFD; (3) analysis of influencing factors for LFD. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were described as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. The Logistic regression method was used for univariate analysis. Results:(1) Clinical data of patients: of the 51 patients, 21 were classified as ordinary type of COVID-19, 19 as severe type and 11 as critical type. In terms of medical history, 31 patients suffered from more than or equal to one kind of chronic disease, 20 had no history of chronic disease. Thirteen patients had the drinking history and 38 had no drinking history. Seven patients were hepatitis positive and 44 were hepatitis negative. Five patients had septic shock at admission, 5 had systemic inflammatory response syndrome (SIRS), and 41 had neither shock nor SIRS. The body mass index (BMI), time from onset to admission, temperature, heart rate, respiratory rate of the 51 patients were (24±3)kg/m 2, (13±5)days, 36.5 ℃ (range, 36.0-38.1 ℃), 82 times/minutes (range, 50-133 times/minutes), 20 times/minutes (range, 12-40 times/minutes). The white blood cell count, level of creatinine, and level of b-type natriuretic peptide within 24 hours after admission were 6.3×10 9/L [range, (2.2-21.7)×10 9/L], 75 μmol/L (range, 44-342 μmol/L), 214 ng/L (range, 5-32 407 ng/L). (2) Analysis of liver function index and treatment of LFD: the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), direct bilirubin (DBil), indirect bilirubin (IBil), activated partial thromboplastin time (APTT) and prothrombin time (PT) were 31 U/L (range, 7-421 U/L), 29 U/L (range, 15-783 U/L), 36 U/L (range, 13-936 U/L), 76 U/L (range, 41-321 U/L), 4.9 μmol/L (range, 2.6-14.3 μmol/L), 5.8 μmol/L (range, 2.6-23.9 μmol/L), 37.2 s (range, 30.9-77.1 s), 13.9 s (range, 12.5-26.7 s), respectively. The percentages of cases with abnormal ALT, AST, GGT, ALP, DBil, IBil, APTT and PT were 47.1%(24/51), 47.1%(24/51), 35.3%(18/51), 13.7%(7/51), 7.8%(4/51), 2.0%(1/51), 21.6%(11/51), and 19.6%(10/51), respectively. Of the 51 patients, LFD was detected in 10 patients classified as ordinary type, in 9 patients as severe type, and in 10 as critical type, respectively. In the 51 patients, 1 of 22 patients with normal liver function developed respiratory failure and received mechanical ventilation within 24 hours after admission, while 9 of 29 patients with abnormal liver function developed respiratory failure and received mechanical ventilation, showing a significant difference between the two groups ( χ2=5.57, P<0.05). (3) Analysis of influencing factors for LFD. Results of univariate analysis showed that clinical classification of COVID-19 as critical type was a related factor for LFD of patients ( odds ratio=10.000, 95% confidence interval: 1.050-95.231, P<0.05). Conclusions:COVID-19 patients with LFD are more susceptible to develop respiratory failure. The clinical classification of COVID-19 as critical type is a related factor for LFD of patients.

This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.
Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis ; Autophagy ; Cell Proliferation ; Cell Line, Tumor ; STAT3 Transcription Factor/metabolism*

This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.
Humans ; Carcinoma, Hepatocellular/genetics* ; Proto-Oncogene Proteins c-akt/metabolism* ; Caspase 3/metabolism* ; Liver Neoplasms/genetics* ; Molecular Docking Simulation ; Sincalide/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Hep G2 Cells ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

Objective: To explore the association between sleep duration and the risk of frailty among the elderly over 80 years old in China. Methods: Using the data from five surveys of the China Elderly Health Influencing Factors Follow-up Survey (CLHLS) (2005, 2008-2009, 2011-2012, 2014, and 2017-2018), 7 024 elderly people aged 80 years and above were selected as the study subjects. Questionnaires and physical examinations were used to collect information on sleep time, general demographic characteristics, functional status, physical signs, and illness. The frailty state was evaluated based on a frailty index that included 39 variables. The Cox proportional risk regression model was used to analyze the correlation between sleep time and the risk of frailty occurrence. A restricted cubic spline function was used to analyze the dose-response relationship between sleep time and the risk of frailty occurrence. The likelihood ratio test was used to analyze the interaction between age, gender, sleep quality, cognitive impairment, and sleep duration. Results: The age M (Q₁, Q₃) of 7 024 subjects was 87 (82, 92) years old, with a total of 3 435 (48.9%) patients experiencing frailty. The results of restricted cubic spline function analysis showed that there was an approximate U-shaped relationship between sleep time and the risk of frailty. When sleep time was 6.5-8.5 hours, the elderly had the lowest risk of frailty; Multivariate Cox proportional risk regression model analysis showed that compared to 6.5-8.5 hours of sleep, long sleep duration (>8.5 hours) increased the risk of frailty by 13% (HR: 1.13; 95%CI: 1.04-1.22). Conclusion: There is a nonlinear association between sleep time and the risk of frailty in the elderly.
Aged ; Humans ; Aged, 80 and over ; Frailty/epidemiology* ; Sleep Duration ; Prospective Studies ; Sleep/physiology* ; China/epidemiology*

Objective: To investigate the effect of hypertensive disorder complicating pregnancy (HDCP) on the mortality and early complications of premature infants. Methods: The general clinical data of preterm infants with gestational age 24-36^{+ 6} weeks were collected from the cooperative units in the task group from January 1, 2013 to December 31, 2014.According to the severity of HDCP, the infants were divided into 4 groups: HDCP group, preeclampsia group, eclampsia group and non HDCP group, the mortality and major complications of preterm infants were compared, and the influencing factors were analyzed. Results: The mortality rate of preterm in the HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=9.970, P=0.019). Eclampsia had a highest fatality rate (4.8%) in the early stage, compared with non HDCP group (2.2%), and the difference was statistically significant.Comparison of HDCP group (1.8%) and eclampsia group (3.2%) suggested that there was no statistically significant difference.The incidence of respiratory distress syndrome (RDS) in preterm in HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=13.241, P=0.004). Eclampsia group showed the highest incidence (35.4%), compared with non HDCP group (16.2%), the difference was statistically significant, but compared with HDCP group (19.9%), preeclampsia group (17.1%), there was no significant diffe-rence.The incidence of bronchopulmonary dysplasia (BPD) in preterm in HDCP group was significantly higher than that of non HDCP group (χ²=9.592, P=0.022), the highest incidence showed up in eclampsia group (9.7%), compared with non HDCP group (2.0%) and HDCP group (1.7%), the difference was statistically significant.But there was no statistically significant difference, compared with preeclampsia group.As the degree of HDCP aggravated, the incidence of BPD gradually rose.There was no significant impact on necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH) and sepsis of HDCP (χ²=7.054, 7.214, 0.358, 3.852; P=0.070, 0.065, 0.949, 0.278). Considering the overall outcome of the child, that was, whether the child died or survived, he had at least one complication, and HDCP had an effect on it (χ²=15.697, P=0.001), so the incidence increased while the degree of HDCP rose gradually.After adjusting gestational age, birth weight, sex, way of delivery, placental abruption and front placenta, prenatal hormonal, gestational diabetes, neonatal asphyxia and other factors, the results displayed that HDCP was the factor leading to the death of premature baby (OR=2.159, 95%CI: 1.093-4.266), and comparison between preeclampsia and eclampsia showed no statistical difference (P=0.714, 0.389); HDCP had no significant influence on RDS, BDP, ICH, NEC, ROP and sepsis. Conclusions HDCP leads to increased risk of premature death, but also leads to the increased incidence of RDS and BPD, but it had no obvious effect on NEC, ROP, IVH, sepsis and other complications.