[Clinical features and lymphocyte subtypes in patients with IgG4-related diseases].

Objective: To deepen understanding of IgG4-related diseases (RDs), we analyzed the associated lymphocyte subtypes, and explored the pathogenesis and potential immunotherapeutic targets. Methods: Eighty-six patients with IgG4-RDs were enrolled, and their clinical characteristics, peripheral lymphocyte subtypes, and disease course were analyzed. Results: The mean age of the participants was 36-87(62±11) years; 51 were male (59.3%) and 35 were women (40.7%); and 34.9% had a history of allergy. Follow-up lasted 4.8 (0.4, 14.1) months. The most common symptoms were abdominal pain, and submandibular gland and lacrimal gland swelling (each 20.9%). Sixty-five (75.6%) participants had multiple organ involvement, and the most frequently affected organs were the pancreas (52.3%), submandibular gland (51.2%), and lacrimal gland (34.9%). A high eosinophil count; high IgE, IgG, IgG1, and IgG4 concentrations; and low complement C3 and C4 concentrations were present in 18.8% (16/85), 30.0% (24/80), 72.9% (62/85), 58.3% (28/48), 89.5% (77/86), 61.2% (52/85), and 50.0% (42/84), respectively, of the participants. In addition, 64.7% (55/85) were positive for autoantibodies, and the most frequent was anti-nuclear antibody (63.5%). The proportion of CD4+T lymphocytes increased in 25.7% (9/35) of the participants, which was accompanied by an increase in the ratio of CD4+/CD8+T lymphocytes (22.9%, 8/35). Importantly, most participants (90.0%, 18/20) had a high proportion of regulatory T (Treg) cells. High interleukin (IL)-2, IL-6, and IL-10 concentrations were present in 50.0% (11/22), 33.3% (10/30), and 16.7% (5/30), respectively, of the participants. Substantial lymphoplasmacytic infiltration, fibrosis, IgG4-positive plasma cell infiltration, and lymphoid follicle hyperplasia or ectopic formation were present in 79.2% (42/53), 67.9%(36/53), 35.8%(19/53) and 30.2% (16/53), respectively, of the participants. Fifty-three participants with detailed pathologic data were also further evaluated, of whom 24.5% (13/53), 3.8% (2/53), and 67.9% (36/53) had definite, probable, and possible diagnoses; and 3.8% (2/53) could not be diagnosed. Compared with baseline, the percentage of eosinophils and the IgE, IgG, and IgG4 concentrations decreased significantly; and the complement C3 and C4 concentrations had increased significantly after 6 months of treatment (all P<0.05). The IgG4 concentration after 6 months of treatment negatively correlated with that of C4, and positively correlated with the baseline concentration of IgE and the IgG4/IgG ratio. Conclusion: IgG4-RDs are a group of diseases characterized by male predisposition; multiple organ involvement; a high eosinophil count; high IgE, IgG, IgG1, and IgG4 concentrations; and a low C3 concentration. Peripheral CD4+T cells and Treg cells are also more abundant. The diseases can be controlled with glucocorticoids and immunosuppressive drugs in the majority of instances. The IgG4 concentration after 6 months of treatment negatively correlates with the baseline complement C4 concentration and positively correlates with the IgE concentration and IgG4/IgG ratio, which suggests that IgG4/IgG, IgE, and complement should be closely monitored to evaluate disease activity and the efficacy of treatment in such patients.

[Single hydrogen-methane breath test for the diagnosis of small intestinal bacterial growth].

Objective: To investigate the diagnostic value of a single hydrogen-methane breath test (SHMBT) for small intestinal bacterial overgrowth (SIBO). Method: The current investigation was a cross-sectional study. Questionnaires and SHMBTs were administered to 162 patients with gastrointestinal symptoms (case group) and 69 healthy volunteers (control group). Differences in SHMBT results between the two groups were assessed,and cut-off values of CH4 (methane) and H2 (hydrogen) were analyzed via receiver operating characteristic (ROC) curves. Lastly,archived SHMBT data from 2 655 patients with gastrointestinal symptoms (validation set) were used to evaluate the diagnostic value of the SHMBT with respect to SIBO. The Chi-square test,the Mann-Whitney U test,Spearman's Rank correlation analysis,and the Z test were used for statistical analysis. Results: Based on the international recommended diagnostic criteria for SIBO,which are fasting CH4 ≥10 ppm (parts per million) or H2 ≥20 ppm,the SHMBT-positive rate in the case group was significantly higher than that of control group (35.2% vs. 21.7%, χ2=4.08, P=0.043). Levels of CH4 and H2 were higher in the case group than in the control group [CH4: 3(2,7) vs. 3(1,3) ppm, H2: 11(4,22) vs. 10(5,15) ppm],and the difference in CH4 levels was statistically significant (Z=6.22,P=0.001). ROC curves were generated based on whether the subjects had gastrointestinal symptoms. The areas under the ROC curves were 0.633 for CH4 alone,0.531 for H2 alone, and 0.620 for CH4 combined with H2. The cut-off values were fasting CH4≥4 ppm,fasting H2≥13 ppm,and fasting CH4 ≥5 ppm (or CH4≥4 ppm and H2≥24 ppm),respectively. Measuring CH4 alone and CH4 combined with H2 was effective for determining the presence of gastrointestinal symptoms (P<0.05). When CH4 alone or CH4 combined with H2 were used as diagnostic indicators of SIBO, the respective SHMBT-positive rates in the validation set were 34.2% and 30.4%. These rates did not significantly differ from the SIBO-positive rate of 32.0% obtained via the international recommended diagnostic criteria (P>0.05). The specificity of CH4 alone was 79.9%,and the accuracy of CH4 alone was 68.8%. The specificity of CH4 combined with H2 was 85.0%,and the accuracy of CH4 combined with H2 was 71.7%. Conclusion: Rapid one-time determination of CH4 and H2 in exhaled breath may a viable diagnostic method for SIBO, and using CH4 combined with H2 (i.e.,fasting CH4≥5 ppm, or CH4 ≥4 ppm and H2 ≥24 ppm) as cutoff values may be feasible.

[Role of CT and MRI image fusion and computer assisted simulation technique in guiding type â ¢ and â £ primary pelvic sarcoma surgeries].

Objective: To explore the possible clinical benefits of CT/MRI image fusion and computer-assisted simulation techniques in guiding type Ⅲ and Ⅳ primary pelvic bone tumor surgeries. Methods: The clinic data of primary bone sarcomas patients treated at Department of Bone and Soft Tissue,Zhenghzhou University Affiliated Cancer Hospital from January 2019 to December 2021 were retrospectively analyzed. Based on whether the CT and MRI image fusion technique was utilized for tumor evaluation and surgical planning,the patients were divided into image fusion group (n=21) or control group (n=27). There were 7 male and 14 female patients included in the image fusion group, with the age of (37.0±10.4) years(range: 18 to 67 years). In the control group, there were 10 males and 17 females with the age of (39.7±15.2) years (range: 16 to 65 years). Both groups included osteosarcoma,chondrosarcoma and undifferentiated polymorphic sarcoma as the pathological diagnosis. Clinical information such as gender,age,pathological diagnosis,location of disease,and metastasis at diagnosis were collected. Surgical related information such as duration of surgery,blood loss,surgical margin,and wound complications were also obtained. Periodical follow-ups every 3 months were performed for all patients to monitor the status of local recurrence,distant metastasis,and survival information. Independent t test and χ² test were used for data comparison between groups. Results: Significant reduced duration of surgery was observed in the image fusion group in comparison with control group both in type Ⅲ and Ⅳ surgeries ((144.0±31.6)min vs. (248.2±56) min,t=-8.084, P<0.01); (173.0±42.0)min vs. (306.1±62.0)min, t=-4.518, P<0.01). Blood loss was significantly reduced in the image fusion group compared with the control group ((484.8±226.3)ml vs. (836.1±359.8)ml,t=-4.130, P<0.01). In addition, significant lower ratio of R1 margin and recurrence rates of type Ⅲ and Ⅳ surgeries were found in the image fusion group comparing with the control group (4.8%(1/21) vs. 22.2%(6/27), χ²=4.214, P=0.040; 4.8%(1/21) vs. 22.2%(6/27), χ²=4.214, P=0.040).In the image fusion group, there were 3 cases of incision infection, 1 of which underwent secondary debridement.And in thecontrol group there were 7 cases of incision infection, 3 of which underwent secondary debridement. There was no significant difference in the incidence of complications between the two groups (14.2%(3/21)vs. 25.9%(7/27), χ²=0.645, P=0.422). Up to the last follow-up, 1 patient died in the image fusion group and 2 patients died in the control group, the difference was not statistically significant (χ²=1.885, P=0.220). Conclusion: Compared with the traditional operation,the image fusion technique can significantly reduce the duration of surgery,blood loss and lower the recurrence rate by achieving better surgical margins.

[Research progress of micronucleus visualization analysis and artificial intelligence detection strategy].

The micronucleomics test can comprehensively display a variety of harmful endpoints, such as DNA damage and repair, chromosome breakage or loss and cell growth inhibition, with fast, simple and economical feature. Micronucleomics is not only widely used in the comprehensive assessment of the types and modes of genetic action of exogenous chemicals (such as drugs, food additives, cosmetics, environmental pollutants, etc.), but also plays an important role in the screening and risk assessment of cancer population at high risk. However, the traditional micronucleomics image counting method has the characteristics of time-consuming, low accuracy, and high cost, which cannot meet the current analysis requirements of large-scale, multi-index, rapidity, high precision and visualization. In recent years, with the rapid development of the era of precision medicine based on big data, visualized analysis of new micronucleomics based on machine learning and detection strategies based on deep learning have shown a good application prospect. This review, based on the application value of micronucleomics, systematically compares the traditional and new artificial intelligence counting of micronucleus images, and discusses the future direction of micronucleus image detection.

[Inhibition of receptor-interacting protein 3 improves experimental autoimmune hepatitis].

Objective: To evaluate the potential of receptor-interacting protein 3 (RIP3) as a therapeutic target for autoimmune hepatitis (AIH). Methods: Immunofluorescence assay was used to observe the activated expression levels of RIP3 and its downstream signal mixed lineage protein kinase domain-like protein (MLKL) in the liver tissues of patients with AIH and hepatic cyst. Concanavalin A (ConA) was injected into the tail vein to induce acute immune-mediated hepatitis in mice. Intervention was performed by intraperitoneal injection of RIP3 inhibitor GSK872 or solvent carrier. Peripheral blood and liver tissues were collected. Serum transaminases level, qPCR and flow cytometry were analyzed. The intergroup comparison was performed with an independent sample t-test. Results: The expression level of p-RIP3 (the activated forms of RIP3) and phosphorylated p-MLKL (MLKL after phosphorylation) downstream signal were significantly higher in the liver tissue of AIH patients than those of controls. Compared with the control group, the expression levels of RIP3 and MLKL mRNA were significantly increased in the liver tissue of AIH patients (relative expression levels 3.28±0.29 vs. 0.98±0.09, 4.55±0.51 vs. 1.06±0.11), and the differences were statistically significant (t=6.71 and 6.77, respectively, and P<0.01). The expression levels of RIP3 and MLKL mRNA were significantly higher in the mice liver tissue of ConA-induced immune hepatitis than those in the control group (relative expression levels 2.35±0.09 vs. 0.89±0.11,2.77±0.22 vs. 0.73±0.16,t=10.4,6.33, P<0.01). RIP3 inhibitor GSK872 had significantly attenuated ConA-induced immune liver injury and inhibited the expression of tumor necrosis factor-α, interleukin-6, interleukin-1ß and NLRP3 in liver. Compared with the control group, the proportions of CD45+F4/80+ macrophages, CD4+ IL-17+ Th17 cells, CD4+ CD25+ regulatory T (Treg) cells and CD11b+ Gr-1+ myeloid derived suppressor cells (MDSCs) were significantly increased in the liver of ConA + Vehicle group. Compared with ConA + Vehicle group, the proportion of CD45+F4/80+ macrophages and CD4+ IL-17+ Th17 cells were significantly decreased, while the proportion of CD4+ CD25+Treg cells and CD11b+ Gr-1+ MDSCs with immunomodulatory functions were significantly increased in mice liver of ConA+GSK872 group. Conclusion: AIH patients and ConA-induced immune hepatitis mice have activated RIP3 signal in liver tissues. Inhibition of RIP3 reduces the expression and proportion of proinflammatory factors and cells, and promotes the accumulation of CD4+ CD25+ Treg cells and CD11b+ Gr-1+ MDSCs with immunomodulatory functions in the liver of mice with immune hepatitis, thereby alleviating liver inflammation and injury. Therefore, the inhibition of RIP3 is expected to be a new approach for the treatment of AIH.

[Albumin-to-bilirubin scores for assessing the prognosis in autoimmune hepatitis-related cirrhosis].

Objective: To investigate the prognostic value of albumin-to-bilirubin scores in the assessment of autoimmune hepatitis-related cirrhosis. Methods: The receiver operating characteristic curve was used to evaluate the accuracy of ALBI, Child-Pugh and model for end-stage liver disease (MELD) for prognosis prediction. Survival analysis was performed according to the ALBI classification. Spearman correlation analysis was performed on the ALBI score and the Child-Pugh score. Survival curves were plotted by Kaplan-Meier method, and Log-rank method was used to compare the survival difference curves between different groups. Results: 149 patients were recruited in the study. The ROC analysis showed that the ALBI scores (0.861, 0.826, 0.779, 0.744)was superior to Child-Pugh scores(0.703, P = 0.006; 0.672, P < 0.001; 0.613, P < 0.001; 0.583, P < 0.001)and MELD score(0.774, P = 0.031; 0.731, P = 0.007; 0.669, P < 0.001; 0.631, P < 0.001) for predicting 6, 12, 24, and 36 months mortality. Patients with ALBI grade 3 had a significantly lower survival rate than those with ALBI grade1 and grade 2. Conclusion: ALBI score may be useful to evaluate the long-term prognosis of patients with autoimmune hepatitis-related cirrhosis.

MicroRNA: a novel biomarker and therapeutic target to combat autophagy in diabetic nephropathy.

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease worldwide and is associated with increased morbidity and mortality in patients with both type 1 and type 2 diabetes. Early treatment of DKD can prevent or slow its progression. Some studies suggest that traditional risk factors such as albuminuria do not effectively predict DKD progression, and other predictors have yet to be characterized and validated. Therefore, there is an urgent need to identify sensitive and easily detectable biomarkers to monitor the decline in renal function. MicroRNAs (miRNAs) have recently emerged as important regulators that are ubiquitous in human tissues and bodily fluids, numerous diseases, including early DKD. Recent developments have revealed that miRNAs-mediated post-transcriptional regulation of gene expression represents an integral part of the autophagy regulatory network. In this review, we explored the utility of miRNAs as biomarkers for the early detection and progression of DKD. We also examined some of the molecular mechanisms by which miRNAs manipulate the autophagic machinery to maintain cellular homeostasis during DKD. A better understanding of the interaction between miRNAs and autophagy may ultimately benefit future DKD diagnosis and therapy.

[Characteristics of initial diagnosis in autoimmune liver disease: an 18-year, retrospective single-center study].

Objective: To analyze the chorological changes of diagnosis in patients with autoimmune liver disease (AILD) and related factors for early diagnosis. Methods: A total of 581 patients with age ranged from 16 to 81 were retrospectively analyzed, who were admitted to Tianjin Medical University General Hospital with AILD during January 2000 to December 2017. Age at diagnosis, diagnostic method and cirrhosis at diagnosis were compared in different groups according to admission period as 2000-2005, 2006-2011, 2012-2017. Results: The diagnostic rate of AILD showed an upward trend during the past near two decades. The proportion of AILD patients diagnosed via health examination was increasing year by year mainly by elevated transaminases (P<0.001). The mean age at diagnosis in our AILD patients were younger at present, especially in men (P=0.044). The proportion of cirrhosis at diagnosis was gradually reduced in three different periods respectively [77.78%(21/27), 41.58% (79/190), 25.00%(91/364), P<0.001], which were coincident in patients with autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) (P<0.001). The shrinking trend of cirrhosis at diagnosis was significantly correlated with the increasing application of health examination (r=-0.549, P<0.001). Conclusions: Extensive application of health examination expands the diagnostic rate of AILD. During the past 18 years, more young patients are diagnosed with AILD. The proportion of severe cases such as cirrhosis at diagnosis is decreasing. Screening of immunological examinations in patients with abnormal transaminases is needed and critical to the early diagnosis of asymptomatic AILD.

MiR-485-5p inhibits metastasis and proliferation of osteosarcoma by targeting CX3CL1.

OBJECTIVE: To investigate the potential effect of miR-485-5p on the development of osteosarcoma (OA) and its relevant mechanism. PATIENTS AND METHODS: The expression level of miR-485-5p was detected in OA tissues and cells (MG-63) comparing with corresponding adjacent normal tissues and normal human osteoblastic cell lines (Hfob1.19), respectively. Luciferase assay was performed to evaluate the interaction between miR-485-5p and CX3CL1, the effects of miR-485-5p on MG-63 cells were determined by subsequent experiments including cell proliferation, expression level of CX3CL1, detection of invasion and migration capacities. RESULTS: In our present research, miR-485-5p was down-regulated in OA tissues and we got the same result in OA cells. In order to obtain potential target of miR-485-5p, we checked it in three publicly available algorithms, TargetScan, miRDB and microRNA. We found that CX3CL1 is a direct target of miR-485-5p, and Luciferase assays confirmed our hypothesis. The results showed that decreased expression of CX3CL1 resulting from the up-regulation of miR-485-5p could decelerate cell proliferation, invasion and migration in OA cells. CONCLUSIONS: We showed the suppressor function of miR-485-5p in OA by targeting CX3CL1, indicating that miR-485-5p/CX3CL1 axis might be a potential therapeutic target for the treatment of OA.

[Standard treatment of autoimmune hepatitis: experience from rheumatoid arthritis].

The treatment of autoimmune hepatitis is similar to that of rheumatic immune disease, which requires the use of hormones and immunosuppressive agents to induce and maintain remission therapy. As one of the most common diseases of rheumatology, rheumatoid arthritis has a definite treatment strategy and gradually becomes a new concept of rheumatoid disease. However, the current treatment of autoimmune hepatitis is still lack of standard compliance treatment strategies, and for the disease activity and immunosuppressive treatment of the efficacy of no uniform standard evaluation criteria, there is no clear evidence of the need to increase the hormone dose or the timing of treatment for patients with substandard treatment, so we consider the standard treatment of autoimmune hepatitis from the experience of rheumatoid arthritis of rheumatism, in order to provide reference for perfecting the standardized treatment of autoimmune hepatitis.

[Role of Rac1 signaling pathway of azathioprine and peptidoglycan in the regulation of monocyte-macrophage apoptosis in Crohn's disease].

Objective: To evaluate the changes of macrophages and expression of Rac1 in the inflammatory site of Crohn's disease, and to investigate the effects of 6-thioguanine (6-TG) and peptidoglycan on apoptosis of human peripheral blood monocyte-macrophage by regulating Rac1 signaling pathway. Methods: Ten patients with Crohn's disease and eight healthy controls diagnosed were enrolled at Department of Gastroenterology and Hepatology, Tianjin Medical University General Hospital from January 2013 to January 2014. The number of macrophages, apoptosis and expression of Rac1 in the inflammation sites and non-inflammation sites of intestinal mucosa were detected in both patients and controls. Peripheral blood mononuclear cells (PBMCs) were sorted by CD14 immunomagnetic beads. The apoptosis of monocytes, expression of Rac1 and related apoptosis signaling molecules were detected in patients treated with peptidoglycan, 6-TG and Rac1 inhibitor NSC23766 and another 15 healthy donors. Results: The number of macrophages and apoptotic cells significantly increased in the inflammatory group of Crohn's disease patients compared with the non-inflammatory group. The expression of PAK1, downstream molecular of Rac1 signaling pathway of macrophages was also significantly higher in the inflammatory group of Crohn's disease patients than that in healthy controls and non-inflammatory group. Compared with control group, anti-apoptotic signals (NF-κB, Bcl-xL and STAT-3) in PBMCs increased in the peptidoglycan group, while slightly decreased in 6-TG group. 6-TG and NSC23766 significantly promoted peptidoglycan-related anti-apoptosis [peptidoglycan group (8.6±3.7)%, peptidoglycan+ 6-TG group (42.0±2.7)%, peptidoglycan+ NSC23766 group (58.5±6.9)%, P<0.05]. Conclusions: Peptidoglycan plays a role in the pathogenesis of Crohn's disease by recruiting macrophages. However, 6-TG inhibits peptidoglycan-induced activation of Rac1 signaling pathway leading to macrophage apoptosis.

[Advances in clinical differentiation between immunological and drug-induced liver injury].

The differentiation between autoimmune hepatitis (AIH) and drug-induced liver injury (DILI) is a difficult task in clinical practice. Some AIH patients had a medication history before disease onset, and some DILI patients may have positive serum antibody. In addition, these two groups of patients have similar clinical symptoms, serological examination results, and liver histopathology, which lead to the difficulties in differentiation. However, correct differential diagnosis is of great significance in making clinical treatment decisions and preventing liver cirrhosis. Therefore, it is necessary to investigate the association between immunological and drug-induced liver injury from the perspectives of pathogenesis, similarities and differences in clinical features, serological examination results, and histological changes, prospects of new biomarkers in differentiation, and the significance of hormone therapy and clinical follow-up in differential diagnosis and treatment, in order to provide a reference for clinical decision-making and research in future.

[The study of liver macrophages polarization in patients with autoimmune hepatitis].

To explore the functional phenotype of liver macrophages in patients with autoimmune hepatitis (AIH). Compared with patients with nonalcoholic fatty liver disease (NAFLD) and chronic hepatitis B (CHB), the absolute CD(68)(+) cell count in patients with AIH was significantly higher (all P<0.05). It was positively correlated with ALT and IgG levels (the correlation coefficients 0.600 and 0.700, P=0.285 and 0.188 respectively). Additionally, compared with patients with NAFLD, the absolute iNOS positive cell count in patients with AIH and CHB were significantly higher (all P<0.05). The expression of TNFα, iNOS and IL-1ß in patients with AIH and CHB were significantly higher than in patients with NAFLD (all P<0.05). Interestingly, compared with patients with AIH and CHB, the absolute CD(206)(+) cell count in patients with NAFLD were significantly higher (all P<0.05). CD(206) expression in patients with NAFLD was higher than patients with AIH and CHB, but with no statistical significance. M1 type macrophages over-expressed and played a major role in the inflammatory reaction and liver injury in patients with AIH.

[Programmed necrosis mediated by receptor-interacting protein 3: a new target for liver disease research].

Hepatocyte death mainly includes apoptosis and necrosis and is a critical process in the pathophysiological mechanism of liver injury caused by various reasons. Recent studies have shown that key regulatory molecules in the inhibition of apoptosis such as caspase cannot be used as targets for inhibiting disease progression in clinical practice. In recent years, programmed necrosis mediated by receptor-interacting protein 3(RIP3)becomes a new hot research topic. It not only plays an important role in inducing inflammatory response, but also is closely regulated by intracellular signal factors, and it is a type of active cell death which can be interfered with. Compared with apoptosis, programmed necrosis is accompanied by the release of various inflammatory factors, which significantly affects local immune microenvironment. RIP3-mediated programmed necrosis has been taken seriously in many diseases. Although its mechanism of action in liver disease remains unclear, the results of recent studies confirmed its important role in the development of liver disease. This article reviews the research advances in the role of RIP3-mediated programmed necrosis signaling pathway in liver disease of various causes and investigates the possibility of RIP3-mediated programmed necrosis as a new target in the treatment of liver disease.