Chronic kidney disease duration and suicide risk among maintenance hemodialysis patients in China.

Background: Our aim was to investigate the relationship between chronic kidney disease (CKD) duration and suicide risk among maintenance hemodialysis patients in China. Methods: Patients with end-stage renal disease (ESRD) who received MHD were enrolled. The demographic and disease characteristics of MHD patients were collected using a self-designed basic information questionnaire. The Suicide Risk Assessment Scale was used to assess suicide risk. Results: A total of 543 (40.8%) patients had suicide risk with Nurses' Global Assessment Scale for Suicide Risk scores ranging from 1 to 19 points. After adjusting for age, gender, disease conditions and mental state, the odds ratios of different CKD duration for suicide risk were 1.00, 2.02, 3.03 and 2.71, respectively (P for trend <.001). There were significant interactions between CKD duration and ESRD duration in relation to suicide risk (P for interaction <.001). There were also interactions between CKD duration and hemodialysis treatment duration, and suicide risk (P for interaction = .01). Patients with ESRD duration of ≤28 months or hemodialysis treatment duration of ≤24 months had the highest risk of suicide when the duration of CKD was 63-94 months, about 2-10 times higher than the other time groups. Conclusions: We found that CKD duration was associated with an increased risk of suicide in maintenance hemodialysis patients in China, independently of other risk factors. Early ESRD and maintenance hemodialysis were associated with suicide in CKD patients.

Bone marrow mesenchymal stem cell-derived exosomes protect podocytes from HBx-induced ferroptosis.

Introduction: Hepatitis B virus-associated glomerulonephritis (HBV-GN) is a common secondary kidney disease in China, the pathogenesis of which is not completely clear, and there is still a lack of effective treatment. Methods: The mechanism of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) was investigated by using HBx-transfected human renal podocytes. Cell viability was detected by CCK8 assay. Iron and malondialdehyde (MDA) contents were detected by using commercial kits. Reactive oxygen species (ROS) levels were measured by flow cytometry analysis. The expression of ferroptosis related molecules was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The effect of miR-223-3p transferred by BMSC-derived exosomes on HBx-overexpressing podocytes was proved by using miR-223-3p inhibitor. Results: The cell viability of podocytes reduced at 72 h or 96 h after the transfection of lentivirus overexpressing HBx protein (p < 0.05). Ferroptosis-related proteins, including glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) were down-regulated upon HBx overexpression, while acyl-CoA synthetase long-chain family member 4 (ACSL4) was up-regulated (p < 0.05). Intracellular levels of iron, MDA, and ROS were also enhanced (p < 0.05). BMSC-derived exosomes protected against ferroptosis induced by HBx overexpression in podocytes. miR-223-3p was enriched in BMSC-derived exosomes. Application of miR-223-3p inhibitor reversed the protective effect of BMSC-derived exosomes on HBx-induced ferroptosis in podocytes. Conclusion: BMSC-derived exosomes inhibit HBx-induced podocyte ferroptosis by transferring miR-223-3p.

HBx-induced PLA2R overexpression mediates podocyte pyroptosis through the ROS-NLRP3 signaling pathway.

INTRODUCTION: Hepatitis B virus-associated glomerulonephritis (HBV-GN) is one of the main types of secondary glomerular diseases, and podocyte injury is an important pathogenic mechanism of HBV-GN, participating in the occurrence and development of HBV-GN. However, the specific mechanism of podocyte injury remains to be studied. METHODS: Human renal podocytes cultured in vitro were divided into six groups. The podocyte morphology was observed under a transmission electron microscope, and the expression of M-type phospholipase A2 receptor (M-PLA2R) on the podocyte membrane was observed by indirect immunofluorescence staining under a fluorescence microscope. The pyroptosis rate and reactive oxygen species (ROS) of podocytes were assessed by FLICA/PI double staining and flow cytometry. Western blot (WB) and quantitative real-time PCR (qPCR) were used to determine the expression of PLA2R, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing card (ASC), caspase-1, IL-1ß, and IL-18. RESULTS: Hepatitis B virus X (HBx) transfected into human renal podocytes in vitro induced the overexpression of PLA2R. Moreover, the overexpressed PLA2R combined with secretory phospholipase A2 group IB (sPLA2-IB) aggravated podocyte injury and increased the pyroptosis rate. In addition, the expression of ROS, the NLRP3 inflammasome and downstream inflammatory factors was increased. In contrast, after inhibiting the expression of PLA2R and ROS, podocyte damage was alleviated, and the pyroptosis rate and the expression of genes related to the ROS-NLRP3 signaling pathway were decreased. CONCLUSION: HBx-induced PLA2R overexpression on the podocyte membrane can significantly upregulate the ROS-NLRP3 signaling pathway, thereby mediating podocyte pyroptosis.

Hepatitis B virus X mediates podocyte pyroptosis by regulating the ROS/NLRP3 signaling pathway in hepatitis B virus-associated glomerulonephritis.

Objectives: This study was designed to investigate whether HBx-induced podocyte injury is related to the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome and the specific mechanism of the oxidative stress pathway in hepatitis B virus-associated glomerulonephritis (HBV-GN). Materials and Methods: The HBx gene was overexpressed in renal podocytes to mimic HBV-GN. Podocyte morphology was observed under a scanning electron microscope. Reactive oxygen species (ROS) generation was detected by dichlorodihydrofluorescein diacetate (DCFH-DA) assay. The podocytes in each group were treated with Hoechst 33342 and subjected to immunofluorescence staining. Caspase-1 activity and LDH levels were assessed with a Caspase-1 Activity Assay Kit and an LDH ELISA Kit, respectively. The expression of all pyroptosis-related proteins was examined by Western blot analysis. Results: Pyroptosis-related proteins, including NLRP3, apoptosis-associated speck-like protein containing card (ASC), caspase-1, IL-1ß, and IL-18 (P<0.05), were up-regulated upon HBx overexpression, and caspase-1 enzyme activity and LDH and Desmin expression were also enhanced (P<0.05). NLRP3 knockdown attenuated the increased expression of pyroptosis-related proteins upon HBx overexpression (P<0.05), which was also achieved by the addition of an ROS inhibitor (P<0.05). Conclusion: HBx regulates podocyte pyroptosis in HBV-GN by targeting the NLRP3 inflammasome, and mitochondrial oxidative stress plays an important role in this process.

MicroRNA-223 downregulation promotes HBx-induced podocyte pyroptosis by targeting the NLRP3 inflammasome.

Hepatitis B virus (HBV) and its related protein, HBV X (HBx), play an important role in podocyte injury in HBV-associated glomerulonephritis (HBV-GN). The microRNA MiR-223 is expressed in several diseases, including HBV-associated disease, while the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome plays a major role in pyroptosis. In this study, we investigated the function and mechanism of action of miR-223 in HBx-induced podocyte pyroptosis. A quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) assay showed that miR-223 was downregulated in HBx-transfected podocytes. Transfection with an miR-223 mimic abolished the expression of the NLRP3 inflammasome and the cytokines that are released as a result of NLRP3 overexpression. Moreover, transfection with HBx and NLRP3 overexpression plasmids increased the expression of pyroptosis-related proteins, especially in the presence of miR-223 inhibitors. Thus, miR-223 downregulation plays an important role in HBx-induced podocyte pyroptosis by targeting the NLRP3 inflammasome, suggesting that miR-223 is a potential therapeutic target for alleviating HBV-GN inflammation.

Chang'an II Decoction ( II )-Containing Serum Ameliorates Tumor Necrosis Factor-α-Induced Intestinal Epithelial Barrier Dysfunction via MLCK-MLC Signaling Pathway in Rats.

OBJECTIVE: To investigate the effect of Chang'an II Decoction ( II ))-containing serum on intestinal epithelial barrier dysfunction in rats. METHODS: Tumor necrosis factor (TNF)-α-induced injury of Caco-2 monolayers were established as an inflammatory model of human intestinal epithelium. Caco-2 monolayers were treated with blank serum and Chang'an II Decoction-containing serum that obtained from the rats which were treated with distilled water and Chang'an II Decoction intragastrically at doses of 0.49, 0.98, 1.96 g/(kg·d) for 1 week, respectively. After preparation of containing serum, cells were divided into the normal group, the model group, the Chang'an II-H, M, and L groups (treated with 30 ng/mL TNF-α and medium plus 10% high, middle-, and low-doses Chang'an II serum, respectively). Epithelial barrier function was assessed by transepithelial electrical resistance (TER) and permeability of fluorescein isothiocyanate (FITC)-labeled dextran. Transmission electron microscopy was used to observe the ultrastructure of tight junctions (TJs). Immunofluorescence of zonula occludens-1 (ZO-1), claudin-1 and nuclear transcription factor-kappa p65 (NF-κ Bp65) were measured to determine the protein distribution. The mRNA expression of myosin light chain kinase (MLCK) was measured by real-time polymerase chain reaction. The expression levels of MLCK, myosin light chain (MLC) and p-MLC were determined by Western blot. RESULTS: Chang'an II Decoction-containing serum significantly attenuated the TER and paracellular permeability induced by TNF-α. It alleviated TNF-α-induced morphological alterations in TJ proteins. The increases in MLCK mRNA and MLCK, MLC and p-MLC protein expressions induced by TNF-α were significantly inhibited in the Chang'an II-H group. Additionally, Chang'an II Decoction significantly attenuated translocation of NF-κ Bp65 into the nucleus. CONCLUSION: High-dose Chang'an II-containing serum attenuates TNF-α-induced intestinal barrier dysfunction. The underlying mechanism may be involved in inhibiting the MLCK-MLC phosphorylation signaling pathway mediated by NF-κ Bp65.

Therapeutic Effect of Chang'an I Recipe ( I ) on Irritable Bowel Syndrome with Diarrhea: A Multicenter Randomized Double-Blind Placebo-Controlled Clinical Trial.

OBJECTIVE: To evaluate the efficacy and safety of Chinese medicine (CM) decoction Chang'an I Recipe ( I ) in the treatment of irritable bowel syndrome with diarrhea (IBS-D). METHOD: A multicenter, randomized, double-blind, placebo-controlled clinical trial was designed. Based on the order of inclusion, the IBS-D patients were randomly assigned to the treatment group or the placebo control group, administrated with Chang'an I Recipe or placebo, 150 mL/bag, 3 times daily, for 8 weeks. The primary indices of efficacy included the effective rates of IBS symptom severity score (IBS-SSS) and the differences in adequate relief (AR) responder; the secondary indexes of efficacy included the changes in scores of the IBS Quality of Life (IBS-QOL) and Hospital Anxiety and Depression (HAD) scales. The safety indices included adverse events and related laboratory tests. RESULTS: A total of 216 patients were included, with 109 in the treatment group and 107 in the control group, and finally 206 were included in the full analysis set (FAS), 191 were included in the per protocol set (PPS). In FAS, the total effective rate was 67.6% and 40.2% for the treatment and control groups, respectively, with 95% confidence interval (CI) for difference in the effective rates between the two groups of 14.4%-40.2%; while in PPS, the total effective rate was 71.3% and 41.2% for the treatment and control groups, respectively (95% CI 16.6%-43.4%). The consistent conclusions of FAS and PPS showed a better efficacy in the treatment group. Both FAS and PPS showed higher AR responder in the treatment group (FAS: 59.6% vs. 35.5%; PPS: 62.8% vs. 38.1%). As for IBS-QOL, the total score and scores in various dimensions of IBS-QOL were not significantly different between the two groups (P>0.05). Both anxiety and depression scales of HAD were not significantly different between the two groups (P>0.05). No adverse events or laboratory abnormalities were found to be obviously related to the tested drugs or clinically significant. CONCLUSION: Chang'an I Recipe was more effective than placebo in the treatment of IBS-D, with no obvious adverse reactions. (No.ChiCTR-TRC-09000328).

A preparation model of Chinese medicine decoction placebo.

Chinese medicine (CM) decoction placebo is one of the key factors restricting the placebo-controlled clinical trial of CM. In this article, based on the analysis of the diffificulties in preparing CM placebo, the production requirements for placebo were put forward. Moreover, in accordance with clinical practices, a production method and evaluation process of CM decoction placebo was proposed, to provide a reference for clinical research.

[Tongjiang granule intervened non-erosive reflux disease patients of Gan-Wei incoordination syndrome: a curative effect observation].

OBJECTIVE: To assess the effectiveness of Tongjiang Granule (TJG) in treating non-erosive reflux disease (NERD) of Gan-Wei incoordination syndrome (GWIS). METHODS: Totally 128 NERD patients of GWIS were recruited from outpatients or inpatients at Department of Digestive Disease, Xiyuan Hospital, China Academy of Chinese Medical Sciences from February 2009 to November 2010. They were randomly assigned to two groups using the block group in the ratio of 1:1, 64 cases in each group. Patients in the experiment group were treated with TJG, 10 g each time, three times a day, while those in the control group were treated with Omeprazole Tablet, 20 mg each time, two times a day. The treatment course of both groups was 4 weeks. The symptoms questionnaires and SF-36 quality of life scale were observed. RESULTS: Finally 114 patients completed the trial. There was statistical difference in epigastric upset, pharyngeal foreign body sensation, abdominal swelling or abdominal pain, and integral of excrement between the two groups before treatment (P < 0.05). However, there was no statistical difference in the rest indices between the two groups (P > 0.05). Compared with before treatment in the same group, the scores of each symptom or the total symptoms were somewhat improved in the two groups (P < 0.01, P < 0.05). There was statistical difference in the rest scores (P > 0.05) except the score of mental health in the experiment group (P < 0.01, P < 0.05). There was statistical difference in the rest scores (P > 0.05) except the score of physical function in the control group (P < 0.01, P < 0.05). There was statistical difference in post-treatment acid reflux, irritability, depression, body pain, roles of emotions (P < 0.01, P < 0.05). The total effective rate was higher in the experimental group, showing no statistical difference when compared with that of the control group (P > 0.05). CONCLUSIONS: TJG had confirmative efficacy in treating NERD patients of GWIS. Meanwhile, it could improve their quality of life, with no obvious adverse reaction.

[Treatment of irritable bowel syndrome by Chinese medicine and pharmacy: an analysis of data mining on experiences of experts].

OBJECTIVE: The pathogenesis of irritable bowel syndrome (IBS), a functional gastrointestinal disorder, is poorly understood. Treatment by Chinese medicine and pharmacy (CMP) is superior to treatment by Western medicine (WM). Therefore, we aimed to analyze Chinese medical experts' experiences in treating IBS by data mining. METHODS: By retrieving related literatures from 1994 to 2012 from CNKI, we chose proved cases and screened effective medical records to establish a database using structural electronic medical case collecting system and data mining. The thinking ways of modern physicians in diagnosis and treatment of IBS were analyzed. RESULTS: Totally 116 medical cases from 46 Chinese medical experts were recruited and analyzed. Results showed that Gan depression and Pi deficiency were most basic syndrome elements. In addition, cold dampness, damp heat, yang deficiency, and Shen deficiency were main elements for the development of diarrhea predominant IBS (IBS-D). Qi stagnation, yin deficiency, qi deficiency, and blood stasis were main elements for the development of constipation predominant IBS (IBS-C). Soothing Gan-qi and strengthening Pi was the most important treatment method. White peony root, prepared atractylodes, prepared licorice, tangerine peel, poria, bupleurum, Radix sileris, lanceolata, and Radix aucklandiae were the most common Chinese herbs used in treating IBS. CONCLUSIONS: (1) Gan depression and Pi deficiency was the main pathogenesis for IBS. Diagnosis and treatment of IBS should be performed clinically according to its development. Tongxie Yao-fang should be used as the basic prescription with modification according to syndrome differentiation. (2) Data mining will have a wide application prospect in studying medical cases.

A randomized, controlled, double-blinded and double-dummy trial of the effect of tongjiang granule on the nonerosive reflux disease of and Gan-Wei incoordination syndrome.

OBJECTIVE: To assess the effectiveness of tongjiang granule (TJG) on the patients with nonerosive reflux disease (NERD) of Gan-Wei incoordination syndrome, its impact on their quality of life, and its safety. METHOD: A randomized, controlled, double-blinded, and double-dummy method was adopted in the trial. There were 120 NERD patients enrolled in the study and randomly divided into the experiment and control groups, each with 60 patients; drugs were distributed according to the drug number by patients' inclusion sequences. In the experiment group, patients were given TJG 10 g and mosapride citrate dummy 5 mg three times a day, and in the control group, patients were given mosapride citrate 5 mg and TJG dummy 10 g three times a day. The treatment courses of both groups were 4 weeks. RESULTS: Among 120 included patients, 112 were screened for full analysis set (FAS), and 105 were screened per-protocol set (PPS). The results were as follows: (1) the improvement of total scores of symptom in the experiment group (0-4 week) were 15.93±7.88 scores by FAS and 16.22 ±7.75 scores by PPS, and they were 10.43±10.16 scores and 10.79±10.27 scores in the control group, respectively. The 95% CI of net scores improvement between the two groups were 2.10-8.90 scores and 1.92-8.94 scores in FAS and PPS; it was significantly better in the experiment group than that in the control group (P<0.05). (2) The improvement of scores of major symptom in the experiment group (0-4 week) were 10.68±5.35 by FAS and 10.89±5.29 by PPS and 7.40±7.41 and 7.60±7.46 in the control group, respectively. The 95% CI of net scores improvement in the two groups were 0.85-5.71 and 0.71-5.69 in FAS and PPS separately, and the improvement in the experiment group was significantly better than that in the control group (P<0.05). (3) The total effective rates were 86.0% and 61.8% in the experiment and the control group separately, and the Ridit analysis results showed that it was better in the experiment group (P<0.05). (4) The improvement quality of life in the domain of physical functioning and general health in the experiment group was better than that in the control group (P<0.05). (5) One case of experiment group caught a cold and recovered in six days without drug suspension. No adverse event was found in the other cases. There was no meaningful safety examination indices change in pretreatment and posttreatment periods in both groups. CONCLUSION: TJG showed a definite effect on the treatment of NERD with Gan-Wei incoordination syndrome, and it could improve the quality of life of NERD patient without obvious toxic and side effects.

[Pay attention to follow-up and monitoring of patients with gastric epithelial dysplasia associated with chronic atrophic gastritis].

Because of the close relationship with stomach cancer and higher rates of carcinoma, gastric epithelial dysplasia associated with chronic atrophic gastritis is considered the most important precancerous lesion of stomach cancer. Due to the absence of recognized treatment methods, periodical follow-up and monitoring were considered an effective method to prevent the occurrence of gastric cancer. Currently, there are numerous areas requiring attention; the authors provide some suggestions on the recognition and key technologies for follow-up and re-examination.