AIM:To determine the effects and mechanisms of interleukin-22 (IL-22) on the fibroblast-like sy-noviocytes ( FLSs) from rheumatoid arthritis ( RA) patients.METHODS:RA-FLSs were cultured by tissue culture meth-od.RA-FLSs were incubated with different concentrations of IL-22 (0,1,10,100μg/L) for 24 h, 48 h and 72 h.The cell viability was examined by CCK-8 assay.IL-22 at concentration of 10 μg/L was used to stimulate RA-FLSs for 24 h, and the change of cell cycle distribution was identified by flow cytometry.The effects of IL-22 at concentrations of 0, 1, 10, 100μg/L and/or STA-21 (a STAT3 inhibitor at concentrations of 0, 25, 50μmol/L) on the protein levels of Bcl-2 and p-STAT3 in the RA-FLSs were determined by Western blot.RESULTS:Compared with control group, stimulation of rhIL-22 at different concentrations for 24 h, 48 h and 72 h, the cells viabilityof RA-FLSs were obviously increased ( P<0.05 ) . After co-cultured with 10 μg/L rhIL-22 for 24 h, the percentages of RA-FLSs were obviously increased in the G2/M+S phase and decreased in the G0/G1 phase.At the same time, rhIL-22 increased, but STA-21 decreased the protein levels of Bcl-2 but p-STAT3 in the RA-FLSs obviously (P<0.05).Treatment with STAT3 inhibitor STA-21 reversed the effect of IL-22-induced Bcl-2 upregulation in the RA-FLSs ( P<0.01 ) .CONCLUSION: STAT3 is critical in the process of IL-22-induced Bcl-2 upregulation in RA-FLSs, indicating that IL-22 may play a role in the apoptosis of RA-FLSs.

Abstract: Objective To establish a rapid detection assay based on fluorescence recombinase polymerase amplification (RPA) targeting Necator americanus eggs, and to evaluate its efficacy, providing technical support for rapid detection of Necator americanus in fecal samples. Methods The fluorescence RPA primers and probe were designed based on the cox1 gene of Necator americanus and then screened the optimal combination to develop the assay. The genomic DNA of Necator americanus eggs was diluted to 7 concentration gradients including 100 pg/µL, 10 pg/µL, 1 pg/µL, 100 fg/µL, 10 fg/µL, 1 fg/µL, 0.1 fg/µL, to determine the detection limit of the assay. The specificity of the assay was demonstrated by detected genomic DNA from Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis and Fasciola hepatica. A total of 44 fecal samples were collected and DNA extraction was performed, and the modified Kato-Katz method, semi-nest PCR method, and fluorescent RPA method were simultaneously used for detection to evaluate the sensitivity and specificity. Results The established fluorescence RPA assay can specifically amplify a fragment of 194 bp of the Necator americanus cox1 gene within 20 min, with a detection limit of 10 fg/µL. There was no cross-reactivity with Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis, Fasciola hepatica after specificity validation. In 44 fecal samples, 27 positive samples were detected by the fluorescence RPA assay, and 26 positive samples were detected by both the Kato-Katz and the semi-nested PCR. The fluorescence curve of sample number 1 was slightly higher than the negative control in the later stage of the reaction, but did not show a similar trend to the positive control, and was therefore judged to be a suspected negative sample. Compared with the Kato-Katz method and the semi-nest PCR method, The sensitivity of the fluorescent RPA method were 100.00% and the specificity were 94.44%, and the consistency of the detection results was good (Kappa=0.953>0.75). Conclusions The assay based on the fluorescence RPA is an efficient, sensitive and specific technique for detecting Necator americanus and it can be applied for surveillance and early warning of hookworm infection.

Abstract: Objective To establish a sensitive and specific nucleic acid detection method for Schistosoma japonicum based on loop-mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR) technology. Methods The LAMP primers, gRNA and ssDNA probe that target Schistosoma japonicum SjR2 genes were designed according to the principles of LAMP and CRISPR. The LAMP-CRISPR reaction system was established and optimized. The sensitivity and specificity of the method were evaluated against the ten-fold serial dilutions of plasmid containing SjR2 target sequences, as well as genomic DNA at different stages of Schistosoma japonicum and other parasites, including Fasciola hepatica, Schistosoma mansoni, Taenia saginata, Clonorchis sinensis, Ascaris lumbricoides, Necator americanus, Paragonimus westermani, and Echinococcus granulosus. Additionally, 15 schistosome-infected snail and 30 uninfected samples were tested by LAMP-CRISPR and LAMP methods, respectively, to evaluate the potential of this method for screening for infected snails. Results　The developed LAMP-CRISPR method was able to specifically amplify and detect the SjR2 gene of S. japonicum. The optimal reaction temperature was 37 ℃, and the optimal reaction concentrations were both 40 nmol/L for gRNA and Cas12a protein. No cross-reaction was observed with genomic DNA from other parasites such as F. hepatica. The detection limit of the method was 10 copies/μL when testing 10-fold dilutions of recombinant plasmids as a template. Furthermore, the LAMP-CRISPR method was able to accurately detect genomic DNA from S. japonicum at various stages of development, including eggs, cercariae, schistosomula, juvenile worms, and adult worms. The results of testing 45 snail samples showed no significant difference between the LAMP-CRISPR and LAMP methods for detecting infected snails (χ2=0.05, P>0.05). The sensitivity and specificity of the LAMP-CRISPR method were 100.00% (15/15) and 96.67% (29/30), respectively, compared to the gold standard, while the sensitivity and specificity of the LAMP method were 100.00% (15/15) and 93.33% (28/30), respectively. Conclusions　This established LAMP-CRISPR detection method presented good sensitivity, specificity and reliability, making it a promising tool for rapid detection and risk monitoring of S. japonicum.

Objective To investigate any potential and independent demographic and serologic risk factors contributing to bone destruction in patients with rheumatoid arthritis ( RA) . Methods A total of 445 patients with RA were recruited in this study. Three autoantibodies including rheumatoid factor ( RF) , anti-cyclic citrullinated peptide antibody ( anti-CCP antibody) and anti-citrullinated alpha-enolase peptide 1 antibody ( anti-CEP-1 antibody) were quantified by using specific ELISA kits. The hand radiographs of all subjects were graded by using the modified Sharp/van der Heijde score ( Sharp score) . The potential and in-dependent risk factors were assessed by using univariate linear regression analyses and the stepwise multiple regression analysis, respectively. Results Based upon the univariate regression analyses, 7 covariates were identified as the potential risk factors for bone destruction in patients with RA, which were female (β=0. 100, P=0. 035), longer disease duration (β=0. 498, P=3. 26×10-29), RF (β=0. 096, P=0. 042), younger age at onset (β=-0. 312, P=1. 60 × 10-11 ), anti-CCP antibody positive (β=0. 202, P=1.74×10-5), anti-CEP-1 antibody positive (β=0.148, P=0.017) and positive for either anti-CCP or anti-CEP-1 antibodies (β=0. 157, P=1. 42×10-3). However, smoking (β=-0. 121, P=0. 018) were identi-fied as the potential protective factors. The multiple regression analysis indicated that the longer disease du-ration (P=2. 24×10-15) and anti-CCP antibody positive (P=0. 012) were independent risk factors for bone destruction. Conclusion Female, longer disease duration, younger age at onset, RF, anti-CCP and anti-CEP-1antibodies are potential risk factors for bone damage in patients with RA. Moreover, longer disease du-ration and anti-CCP antibody are two independent risk factors contributing to bone destruction in RA.

BACKGROUND:Perioperative high hidden blood loss affects the recovery of joint function after total hip replacement. OBJECTIVE:To analyze the reliability of the Mini Nutritional Assessment on evaluating the nutritional status in elderly patients with femoral neck fracture on admission, and to investigate the effect of nutritional status variation on hidden blood loss after total hip replacement. METHODS:234 elderly patients with femoral neck fracture underwent total hip replacement. By using Mini Nutritional Assessment, patients were randomly divided into three groups:wel-nourishment group (n=52), malnourishment at risk group (n=92), and malnourishment group (n=90). The results were used to analyze the correlation of Mini Nutritional Assessment and serological nutrition indicators, and to hidden blood loss. RESULTS AND CONCLUSION:(1) Hidden blood loss:101 patients suffered from high hidden blood loss. Hidden blood loss, its proportion to total blood loss and incidence of high hidden blood loss gradual y increased with the deterioration of the nutritional status (P<0.05). (2) Mini Nutritional Assessment:Pre-operative Mini Nutritional Assessment score, and the incidence of hidden blood loss evaluated by albumin, prealbumin, transferrin, lymphocyte count, the percentage of lymphocytes and hemoglobin was significantly higher in patients with high hidden blood loss than those with low hidden blood loss (P<0.01). (3) Results of correlation analysis:High hidden blood loss was positively correlated with pre-operative Mini Nutritional Assessment, albumin, prealbumin, transferrin, the percentage of lymphocytes and hemoglobin (P<0.05). (4) These findings confirm that risk evaluation with Mini Nutritional Assessment is a reliable method to assess the nutritional status in elderly patients undergoing total hip replacement. Its combination with various serum nutrition indicators can determine high hidden blood loss and the prognosis.

OBJECTIVETo investigate the effects of initiative and passive perioperative function exercises on hidden blood loss (HBL).

METHODSTwo hundreds and thirty elderly patients with hip fractures aging from 67 to 87 years (average age of 73.6 years) who underwent total hip replacement were included. By the intensity and the manner of perioperative function exercises, patients were divided into four groups: little initiative function exercises group (group A, n=51), little initiative and passive function exercises group (group B, n=54), normal initiative function exercises group (group C, n=65), normal initiative and passive function exercises group (group D, n=60). The true total blood loss, HBL and their proportion on the original blood volume and total blood loss was calculated depending on height, weight, intra-operative blood loss, post-operative blood loss, pre- and post-operative hematocrit, and blood transfused. According to the proportion of mean HBL on total blood loss, patients were divided into low HBL group and high HBL group. The data were analyzed by t test.

RESULTSThe mean HBL was 517 ml, 41.9% of the total blood loss. Thereinto, the mean HBL was 695 ml in group A, 49.3% of the total blood loss, the prevalence of high HBL was 66.7% (34/51); the mean HBL was 625 ml in group B, 46.9% of the total blood loss, the prevalence of high HBL was 59.3% (32/54); the mean HBL was 446 ml in group C, 38.4% of the total blood loss, the prevalence of high HBL was 30.8% (20/65); the mean HBL was 346 ml in group D, 32.3% of the total blood loss, the prevalence of high HBL was 20.0% (12/60). Mean HBL, mean HBL/total blood loss, prevalence of high HBL were lower in group C than that in group A and group B (all P<0.05); and were lower in group D than that in group C (all P<0.05). The prevalence was 57.4% (132 cases) in low HBL group, and 42.6% (98 cases) in high HBL. The proportion of little initiative function exercises patients in high HBL group was obviously higher than that in low HBL group (P<0.05).

CONCLUSIONSThe intensity and the manner of perioperative function exercises are strongly associated with the HBL in elderly patients with total hip replacement. The initiative combined with the passive function exercises could be effectively prevent and reduce the incidence of high HBL.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Blood Loss, Surgical ; Exercise Therapy ; adverse effects ; Hip Fractures ; surgery ; Humans ; Postoperative Hemorrhage

More and more studies have shown that nonalcoholic fatty liver disease (NAFLD) is closely associated with chronic kidney disease (CKD). Although the causal relationship has not been clarified, the mechanisms, such as metabolic disorders, diet-intestinal microbiota axis disorders, oxidative stress, platelet activation, and genetic and epigenetic regulation, play an important role in the disease interaction network. This article reviews the evidence for the association between NAFLD and CKD and the potential mechanisms under which NAFLD and its related factors may increase the risk of CKD, and moreover, it proposes related patient management recommendations and treatment strategies.

Objective:To investigate the current situation of motivations for hospice care volunteerism among undergraduate nursing students, and to analyze its influencing factors.Methods:Using the convenient sampling method, a total of 575 nursing students from Nursing Department of Xinjiang Medical University were selected as the research objects in August 2022. They were investigated using general information questionnaire, Chinese version of Inventory of Motivations for Hospice Palliative Care Volunteerism Scale, The Palliative Care Quiz for Nursing, the Organizational Climate Scale and Prosocial Tendencies Measure Scale.Results:The score of Motivations for Hospice Palliative Care Volunteerism Scale for 575 nursing students was (87.32±20.54) , score of Palliative Care Quiz for Nursing was (7.89±3.44) , score of Organizational Climate Scale was (49.35±10.33) and the score of Prosocial Tendencies Measure scale was (96.71±18.25) . Qualities and abilities that college student volunteers should possess, access to hospice care, pro-social tendencies, and knowledge of palliative care were influential factors in nursing students' motivation to volunteerism for hospice care ( P<0.05) , which could explain 19.9% of the total variation. Conclusions:The motivations for hospice care volunteerism of undergraduate nursing students is in a medium and above level. Nursing educators should strengthen education and training related to nursing students' participation in hospice care, so that nursing students will participate in voluntary service activities through their own professional advantages, strengthen the voluntary team of hospice care, and promote the development of voluntary hospice care services.

Objective:To analyze the association of two single-nucleotide polymorphisms (SNP) [Calcitonin gene related peptide (CGRP) rs155209 and receptor activity modifying protein 1 (RAMP1) rs3754701] and the prognosis of chronic hepatitis B patients who were under interferon therapy.Methods:A total of 317 patients and their anticoagulant blood samples were collected in this study. The SNPs in the CGRP and region RAMP1 were genotyped using matrix-assisted laser desorption/ionization time of flight mass spectrometry. Logistic regression method was used to assess the results from different phenotypic outcomes between cases and controls, after adjusted for sex and age in co-dominant, dominant and recessive genetic models.Results:Data from this study clearly demonstrated the relevance of CGRP rs155209 and RAMP1 rs3754701 with DNA response and ALT response. RAMP1 rs3754701T was strongly associated with both DNA response and ALT response ( OR=2.277, 95 %CI: 1.386-3.741, P=0.001; OR=1.694, 95 %CI: 1.073-2.675, P=0.024). However, CGRP rs155209C was less prone to DNA response and ALT response ( OR=0.150, 95 %CI: 0.083-0.271, P<0.001; OR=0.583, 95 %CI: 0.367-0.925, P=0.022). Conclusions:Results from our study suggested that both RAMP1 rs3754701 and CGRP rs155209 were associated with the prognosis of patients under interferon therapy in Han population, from the northern parts of China while RAMP1 rs3754701T was a protective factor for both ALT response and DNA response, but CGRP rs155209C carriers were less prone to DNA and ALT responses.

Objective Find abnormal changes of plasma lipid metabolism-related proteins before 20 weeks of gestation in patients with hypertensive disorder of pregnancy(HDP), and preliminarily investigate the role of plasma apolipoprotein C4 elevation in HDP. Methods A nested case-control study was used. The plasma were collected from pregnant women who underwent routine prenatal examination in Guangzhou Women and Children's Medical Center from November 2014 to March 2017. Label-free mass spectrometry was used to detect the differences in plasma lipid metabolism-related proteins before 20 weeks of gestation between 12 pairs of HDP patients and normal controls, and different 48 pairs of samples were used for verification. The protein with the most significant difference multiples was screened to study its effects on monolayer permeability and nitric oxide secretion of endothelial cells. One-way ANOVA was used for comparison between groups, and P<0.05 was considered as statistically significant difference. Results Compared with the control, the lipid metabolism-related proteins, APOC4, Fatty acid-binding protein 4 (FABP4), Apolipoprotein E (APOE), Apolipoprotein C3 (APOC3) and Beta-2-glycoprotein 1(APOH) raised to 1.94, 1.82, 1.59, 1.55 and 1.38 times, phospholipid transfer protein (PLTP) decreased to 0.78 times in plasma before 20 weeks of pregnancy of patients with HDP (t value were 2.499, 2.497, 2.081, 2.098, 2.426 and 2.564, respectively, P<0.05). Cell experiments results showed that 50 ng / ml APOC4 significantly increased 20％ HUVEC single layer cell permeability to FITC-labeled dextran (F=455.4, P<0.01), and significantly decreased the level of nitric oxide in the supernatant of HUVEC culture by 25％ (F=61.92, P<0.01). Conclusions Before diagnosis, plasma protein levels involved in lipid metabolism in HDP patients have been changed, resulting in abnormal lipid metabolism. APOC4 can increase the permeability of vascular endothelial cells, inhibit endothelial source of NO secretion, cause endothelial dysfunction.