Deficiency in glutathione synthesis and reduction contributes to the pathogenesis of colitis-related liver injury / 中南大学学报(医学版)

OBJECTIVES@#Liver disease is the most common extra-intestinal manifestation of ulcerative colitis (UC), but the underlying pathogenesis is still not clarified. It is well accepted that the occurrence of UC-related liver disease has close correlation with immune activation, intestinal bacterial liver translocation, inflammatory cytokine storm, and the disturbance of bile acid circulation. The occurrence of UC-related liver disease makes the therapy difficult, therefor study on the pathogenesis of UC-related liver injury is of great significance for its prevention and treatment. Glutathione (GSH) shows multiple physiological activities, such as free radical scavenging, detoxification metabolism and immune defense. The synthesis and the oxidation-reduction all contribute to GSH antioxidant function. It is reported that the deficiency in hepatic GSH antioxidant function participates in multiple liver diseases, but whether it participates in the pathogenesis of UC-related liver injury is still not clear. This study aims to investigate the feature and underlying mechanism of GSH synthesis and oxidation-reduction function during the development of UC, which will provide useful information for the pathogenesis study on UC-related liver injury.@*METHODS@#UC model was induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS)-ethanol solution (5 mg/0.8 mL per rat, 50% ethanol) via intra-colonic administration in rats, and the samples of serum, liver, and colon tissue of rats were collected at the 3rd, 5th, and 7th days post TNBS. The severity degree of colitis was evaluated by measuring the disease activity index, colonic myeloperoxidase activity, and histopathological score, and the degree of liver injury was evaluated by histopathological score and the serum content of alanine aminotransferase. Spearman correlation analysis was also conducted between the degree of colonic lesions and index of hepatic histopathological score as well as serum aspartate aminotransferase level to clarify the correlation between liver injury and colitis. To evaluate the hepatic antioxidant function of GSH in UC rats, hepatic GSH content, enzyme activity of GSH peroxidase (GSH-Px), and GSH reductase (GR) were determined in rats at the 3rd, 5th, and 7th days post TNBS, and the protein expressions of glutamine cysteine ligase (GCL), GSH synthase, GSH-Px, and GR in the liver of UC rats were also examined by Western blotting.@*RESULTS@#Compared with the control, the disease activity index, colonic myeloperoxidase activity, and histopathological score were all significantly increased at the 3rd, 5th, and 7th days post TNBS (all P<0.01), the serum aspartate aminotransferase level and hepatic histopathologic score were also obviously elevated at the 7th day post TNBS (all P<0.05). There was a significant positive correlation between the degree of liver injury and the severity of colonic lesions (P=0.000 1). Moreover, compared with the control, hepatic GSH content and the activity of GSH-Px and GR were all significantly decreased at the 3rd and 5th days post TNBS (P<0.05 or P<0.01), and the protein expressions of GCL, GSH-Px, and GR were all obviously down-regulated at the 3rd, 5th, and 7th days post TNBS (P<0.05 or P<0.01).@*CONCLUSIONS@#There is a significant positive correlation between the degree of liver injury and the severity of colonic lesions, and the occurrence of reduced hepatic GSH synthesis and decreased GSH reduction function is obviously earlier than that of the liver injury in UC rats. The reduced hepatic expression of enzymes that responsible for GSH synthesis and reduction may contribute to the deficiency of GSH synthesis and oxidation-reduction function, indicating that the deficiency in GSH antioxidant function may participate in the pathogenesis of UC related liver injury.

Multicenter study of the clinicopathological features and recurrence risk prediction model of early-stage breast cancer with low-positive human epidermal growth factor receptor 2 expression in China (Chinese Society of Breast Surgery 021) / 中华医学杂志(英文版)

BACKGROUND@#: Breast cancer with low-positive human epidermal growth factor receptor 2 (HER2) expression has triggered further refinement of evaluation criteria for HER2 expression. We studied the clinicopathological features of early-stage breast cancer with low-positive HER2 expression in China and analyzed prognostic factors.@*METHODS@#: Clinical and pathological data and prognostic information of patients with early-stage breast cancer with low-positive HER2 expression treated by the member units of the Chinese Society of Breast Surgery and Chinese Society of Surgery of Chinese Medical Association, from January 2015 to December 2016 were collected. The prognostic factors of these patients were analyzed.@*RESULTS@#: Twenty-nine hospitals provided valid cases. From 2015 to 2016, a total of 25,096 cases of early-stage breast cancer were treated, 7642 (30.5%) of which had low-positive HER2 expression and were included in the study. After ineligible cases were excluded, 6486 patients were included in the study. The median follow-up time was 57 months (4-76 months). The disease-free survival rate was 92.1% at 5 years, and the overall survival rate was 97.4% at 5 years. At the follow-up, 506 (7.8%) cases of metastasis and 167 (2.6%) deaths were noted. Multivariate Cox regression analysis showed that tumor stage, lymphvascular invasion, and the Ki67 index were related to recurrence and metastasis (P < 0.05). The recurrence risk prediction model was established using a machine learning model and showed that the area under the receiving operator characteristic curve was 0.815 (95% confidence interval: 0.750-0.880).@*CONCLUSIONS@#: Early-stage breast cancer patients with low-positive HER2 expression account for 30.5% of all patients. Tumor stage, lymphvascular invasion, and the Ki67 index are factors affecting prognosis. The recurrence prediction model for breast cancer with low-positive HER2 expression based on a machine learning model had a good clinical reference value for predicting the recurrence risk at 5 years.@*TRIAL REGISTRATION@#: ChiCTR.org.cn, ChiCTR2100046766.

Clinical efficacy of laparoscopic sleeve gastrectomy in treatment of obesity associated nonal-coholic fatty liver disease / 中华消化外科杂志

Objective:To investigate the clinical efficacy of laparoscopic sleeve gastrec-tomy (LSG) in the treatment of obesity associated nonalcoholic fatty liver disease (NAFLD).Methods:The retrospective and descriptive study was conducted. The clinical data of 40 patients with obesity associated NAFLD who were admitted to Xijing Hospital of Digestive Diseases of Air Force Medical University from January 2018 to June 2019 were collected. There were 19 males and 21 females, aged (27±8)years. Patients underwent LSG after preoperative examinations and evaluations. Observation indicators: (1) surgical and postoperative conditions; (2) follow-up. Follow-up was conducted by telephone interview, WeChat, and outpatient review to detect the liver function, glucose and lipid metabolism-related status as well as the relief of comorbidities of patients at 6 and 12 months after LSG. The follow-up was up to June 2020. Measurement data with normal distribu-tion were represented as Mean± SD. Repeated measurement data were analyzed by the repeated measures ANOVA. Count data were represented as absolute numbers, and chi-square test was used for comparison between groups. Results:(1) Surgical and postoperative conditions: 40 patients underwent LSG successfully, without intraoperative conversion to laparotomy or death. The operation time, volume of intraoperative blood loss, and duration of postoperative hospital stay were (83±12)minutes, (19±7)mL and (3.7±0.7)days. Among the 40 patients, 1 case had fat liquefac-tion after LSG and 2 had abdominal distension within a month, who were improved after symp-tomatic and supportive treatment. (2) Follow-up: 40 patients were followed up at postoperative 6 and 12 months. The body mass of 40 patients was (112±17)kg, (85±16)kg, (73±11)kg before operation, at postoperative 6 and 12 months, respectively. The body mass index was (39.7±4.2)kg/m 2, (30.0±4.5)kg/m 2, (25.9±2.9)kg/m 2 before operation, at postoperative 6 and 12 months, respectively. The alanine aminotransferase was (56.40±39.51)U/L, (30.15±17.93)U/L, (26.45±9.44)U/L before operation, at postoperative 6 and 12 months, respectively. The aspartate amino-transferase was (34.03±19.62)U/L, (28.20±10.27)U/L, (24.90±8.44)U/L before operation, at post-operative 6 and 12 months, respectively. The gamma-glutamyltransferase was (48.68±28.45)U/L, (43.30±18.56)U/L, (32.80±12.09)U/L before operation, at postoperative 6 and 12 months, respectively. The fasting blood-glucose was (6.51±2.38)mmol/L, (5.64±1.51)mmol/L, (4.98±0.91)mmol/L before operation, at postoperative 6 and 12 months, respectively. The triglyceride was (1.81±0.95)mmol/L, (1.48±0.57)mmol/L, (1.17±0.44)mmol/L before operation, at postoperative 6 and 12 months, respectively. The total cholesterol was (4.77±0.98)mmol/L, (4.58±0.75)mmol/L, (4.10±0.68)mmol/L before operation, at postoperative 6 and 12 months, respectively. The low density lipoprotein cholesterin was (2.99±0.81)mmol/L, (2.79±0.54)mmol/L, (2.48±0.40)mmol/L before operation, at postopera-tive 6 and 12 months, respectively. The high density lipoprotein cholesterin increased was (1.08±0.20)mmol/L, (1.15±0.18)mmol/L, (1.45±0.21)mmol/L before operation, at postoperative 6 and 12 months, respectively. Compared with preoperative indicators, there were significant differences in postoperative body mass, body mass index, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, fasting blood-glucose, triglyceride, total cholesterol and low density lipoprotein cholesterin ( F=472.439, 533.730, 26.791, 11.029, 25.545, 27.994, 27.534, 36.220, 18.596, P<0.05), showing a downward trend. There was a significant difference between preoperative and postoperative high density lipoprotein cholesterin ( F=68.974, P<0.05), showing a upward trend. The excess weight loss of 40 patients at 6 months and 12 months after LSG were 61%±18% and 85%±15%. The preoperative ultrasonography of 40 patients showed no, mild, moderate and severe fatty liver in 0, 2, 15 and 23 cases, respectively. The above indicators were 8, 9, 12, 11 cases at post-operative 6 months, and 23, 11, 6, 0 cases at postoperative 12 months, respectively. There was a significant difference of the grade of NAFLD between patients before and after surgery ( χ2=68.210, P<0.05). Conclusion:LSG has an accurate short-term clinical efficacy for obesity associated NAFLD, which can reduce body mass, improve liver function, alleviat the disorders of glucose and lipid metabolism and the severity of medium-severe fatty liver.

Myeloid-specific targeting of Notch ameliorates murine renal fibrosis via reduced infiltration and activation of bone marrow-derived macrophage

Macrophages play critical roles in renal fibrosis. However, macrophages exhibit ontogenic and functional heterogeneities, and which population of macrophages contributes to renal fibrosis and the underlying mechanisms remain unclear. In this study, we genetically targeted Notch signaling by disrupting the transcription factor recombination signal binding protein-Jκ (RBP-J), to reveal its role in regulation of macrophages during the unilateral ureteral obstruction (UUO)-induced murine renal fibrosis. Myeloid-specific disruption of RBP-J attenuated renal fibrosis with reduced extracellular matrix deposition and myofibroblast activation, as well as attenuated epithelial-mesenchymal transition, likely owing to the reduced expression of TGF-β. Meanwhile, RBP-J deletion significantly hampered macrophage infiltration and activation in fibrotic kidney, although their proliferation appeared unaltered. By using macrophage clearance experiment, we found that kidney resident macrophages made negligible contribution, but bone marrow (BM)-derived macrophages played a major role in renal fibrogenesis. Further mechanistic analyses showed that Notch blockade reduced monocyte emigration from BM by down-regulating CCR2 expression. Finally, we found that myeloid-specific Notch activation aggravated renal fibrosis, which was mediated by CCR2 macrophages infiltration. In summary, our data have unveiled that myeloid-specific targeting of Notch could ameliorate renal fibrosis by regulating BM-derived macrophages recruitment and activation, providing a novel strategy for intervention of this disease.

Behavioral problems of HIV infected children / 中国学校卫生

Objective@#To understand the behavioral problems of children infected with HIV, and to provide reference for further psychological intervention.@*Methods@#The survey was conducted from January to December 2017. A total of 256 HIV-infected 10-16 years old school-aged children who were receiving highly active antiretroviral therapy (HAART) in Guangxi Zhuang Autonomous Region Center for Disease Control and Prevention were invited to participate. Another 256 children without HIV infection who were matched with sex and age were selected as the control group. Achenbach Children’s Behavior Scale was used to conduct psychological questionnaires in the two groups of children.@*Results@#Activity, social competence, and learning ability in the study group scored lower than control group (P<0.05). In children with HIV infection group, score of social competence in boys and girls aged 10-11 years were (14.35±3.96) (15.26±5.00), respectively, and was (14.21±4.64) and (14.57±5.50) in boys and girls aged 12-16 years was significantly lower than age and sex-matched control group (19.38±6.77 and 18.09±5.16 in boys and girls aged 10-11 years, 17.26±6.88 and 17.84±6.76 in boys and girls aged 12-16 years) (P<0.05). The total detection rate of behavioral problems in the study group (13% and 10% in boys and girls aged 10-11 years, 17% and 13% in boys and girls aged 12-16 years) was significantly higher than that in the control group (5% and 3% in boys and girls aged 10-11 years, both 4% in boys and girls aged 12-16 years) (P<0.05).@*Conclusion@#Behavioral problems in HIV-infected children are prevalent, which worth more attention to carry out psychological intervention among this vulnerable population.

Research progress of deubiquitinating enzyme CYLD to regulate liver-related diseases / 中华肝脏病杂志

Cylindromatosis gene is a kind of tumor suppressor genes, whose mutation or deletion will lead to the development of a cylindrical tumor. The deubiquitinating enzyme CYLD protein encoded by it is a member of the deubiquitinating enzyme family. CYLD alters the function of the target molecules by removing the ubiquitin chain linked to the substrate protein K63, and participates in the regulation of signaling pathways, such as NF-κB, JNK and Wnt. This article reviews the recent year’s research progress of CYLD, especially its negative regulatory role in the progression of liver-related diseases.

Impact of post-intensive care syndrome of ICU survivors on burden of primary caregivers / 中国实用护理杂志

Objective To explore the impact of post-intensive care syndrome (PICS) of ICU survivors on burden of primary caregivers. Methods This study included ICU survivors and primary caregivers who meeting the conditions between 2016.4.1 and 2017.6.30. Mini-mental State Examination, The Hospital Anxiety and Depression Scale, Post-traumatic Stress Disorder Self-rating Scale and the Barthel Index Rating Scale were used to assess PICS in ICU survivors at 3 months after discharge from ICU. At the same time, the Chinese Zarit Burden Interview was used to assess the primary caregivers ' burden. Univariate and multivariate logistic regression were used to analyze the relationship between PICS and primary caregiver burden. Results PICS occurred in 127 patients. 228 (58.9%) caregivers had mild or no burden, 102 (26.4% ) had moderate burden and 57 (14.7% ) had severe burden. Logistic multiple regression analysis showed that the patient's PICS was an independent risk factor for the caregiver's burden [ OR=1.547(1.143-6.189), P=0.018]. Conclusions PICS of ICU survivors can increase the burden of primary caregivers. Necessary PICS education for patients and caregivers may have a certain significance to reduce the level of burden.

Effect of macrophage polarization on interstitial fibrosis in mouse unilateral ureteral obstruction model / 中华肾脏病杂志

Objective To investigate the effect of macrophage polarization on tubulointerstitial fibrosis of mouse unilateral ureteral obstruction(UUO) model.Methods Twelve male C57BL/6J mice were employed,each of which with an age of 8 to 10 weeks.UUO model was established with these mice with the method of unilateral ureteral ligation.Mice were then sacrificed on the 7th and 14th day respectively after operation,and renal tissue specimens were obtained.The authors detected collagen deposition by Masson staining,and alpha smooth muscle actin (alpha SMA) as well as collagen type Ⅰ (Coll-1) mRNA by real-time quantitative PCR.The authors also detected the degree of renal interstitial macrophages infiltration and expression changes of polarization by immunofluorescence staining.Results Compared with the mice that were observed on the 7th day after operation,the degree of renal interstitial fibrosis in mice observed on the 14th day after operation was comparatively serious,the difference shown by semi-quantitative results was statistically significant (P ＜ 0.05).Moreover,mice observed on the 14th day after operation have more M2 macrophages,the difference between two groups of mice was statistically significant (P ＜ 0.05).On the contrary,there was no statistically significant difference in the degree of M1 macrophages infiltration between these two groups of mice.Conclusions In the renal interstitial fibrosis model induced by UUO,the degree of macrophage infiltration increased significantly,mainly resulted from M2 macrophage infiltration,suggesting that M2 macrophages were involved in the formation of renal fibrosis.

Study of pharmacokinetics of digoxin in ovariectomized rats model / 药学学报

This study aims to investigate the change of plasma concentration of digoxin (DIG) in rats with ovariectomy. Twelve female SD rats were randomly assigned into ovariectomized group and sham group (n = 6). All rats plasma was collected after a single dose of 2 mg x kg(-1) DIG administrated orally, serum DIG concentration was determined by LC-MS/MS. The level of P-gp in the intestinal was analyzed by Western blotting. Pharmacokinetic calculations were performed on each individual using DAS 2.0 practical pharmacokinetic software. Compared with the sham group, C(max) of ovariectomized group decreased significantly (P < 0.01). There was no significant difference of AUC(0-t), and the level of P-gp was elevated in ovariectomized group. It was found that C(max) of DIG was significantly reduced after ovariectomy, and the change was associated with the decreased level of estrogen, which contributes to the increased level of P-gp.

The expression of protein fused HCV envelope protein E2 with His tag and its implication / 解放军医学杂志

Objective To construct the eukaryotic expression vector coding HCV gene E2 fused with His-Tag, and to express fused protein in CHO cells for investigating the function of HCV envelope protein E2. Methods The gene encoding HCV envelope protein E2 was amplified from pBRTM/HCV1-3011, a plasmid containing the cDNA of HCV's ORF, by polymerase chain reaction (PCR) method and cloned into the vector pET28(a) containing His-Tag to obtain the fused HCV envelope protein E2 gene fused with His-Tag. The fused gene was cloned into pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-His-E2, which will express the E2 protein, fused with His tag. This recombinant plasmid was transfected into CHO cells by Lipofactamine 2000 reagent. The fused protein was identified by indirect immunofluorescence (IIF) and Western-blot (WB) methods. Result The positive results were obtained when the fused protein of HCV E2 with His-Tag were identified by IIF and WB methods. Conclusion The eukaryotic expression vector pcDNA3.1-His-E2 was constructed successfully and the fused proteins were expressed in cells.