[Objective] This paper sums up Professor Ye Bai's scholar thought and clinical experience of using jineijin couplet medicines in treating digestive disease. [Methods] From the aspects of the drug pairing, application characteristics and case examples, this paper sums up Professor Ye Bai's scholar thought and clinical experience of using jineijin couplet medicines in treating digestive disease. [Results] Jineijin and ezhu can remove blood stasis and promote di-gestion to help transportion; jineijin and haijinsha can clear heat and dampness and not only treat jaundice, but also remove stones.[Conclusions]Professor Ye Bai's scholar thought and clinical experience of using jineijin couplet medicines in treating digestive disease is effective and is trusted and welcomed by patients. His thought and clinical experience are worth to extend and apply.

Objective To investigate the roles of Arsenic Trioxide to the lupus nephritis in MRL/ lpr mice and the effect on the expression of TGF-pl in the renal tubule. Methods 45 MRL/lpr mice were chosen for such experiment, then separated in 3 different groups, including arsenic trioxide ( ATO) group, cyclophosphamide (CYC) group and sodium chloride (NS) group. Urine of these mice was kept after the treatment to detect the urine protein levels. The pathological changes and the expression of IgG were observed , and the complement 3 of the nephridial tissue as well as the TGF-pl were detected by immunohisto-chemistry. Result The levels of urine protein in ATO group and CTX group were lower than NS group after the treatment ( P <0.05). The expression of IgG along the glomerular mesangium and capillary loop in ATO and CTX groups was less than NS group ( P <0.05). The expression of C3 had no significant difference within three groups ( P > 0.05). The glomcrulus spherulous cells and the integral of activity in ATO group and CTX group were less than that in NS group ( P <0.05). TGF-β1 wildly presented in the epithelial cells endochylema of the nephric tubule, but it was rarely seen in glomcrulus in NS group. The expression in the nephric tubule of ATO group was less than NS group (20.28 ± 1. 90 vs 68. 23 ± 2. 87, P < 0.01) , and the expression in the nephric tubule in CTX group was less than that in NS group (23. 26 ± 1.71 vs 68. 23 ±2. 87, P <0. 01). Conclusion TGF-pl may take part in the immunopathogenesis of lupus nephritis in MRI/lpr mice. ATO can relieve the inflammation of nephric tubule by down regulation the expression of TGF-β1. ATO can also relieve the tissue damage of kidney in lupus nephritis through reducing the inflammation of glomcrulus and interstitial substance,as well as the expression of the immunocomplex.

Objective:To prepare the polyclonal antibodies against advanced oxidation protein products (AOPP),and to provide an effective agent for research on the pathogenesis of AOPP and assess exactly the relationship between AOPP and relative diseases.Methods:AOPP-rabbit serum albumin (AOPP-RSA) was prepared by treating RSA with hypochloric acid.The rabbit anti-AOPP-RSA polyclonal antibodies were generated and purified by affinity chromatography. The titers and the specificity of the antibodies were measured by ELISA.The plasma AOPP and the localization of AOPP in nephridial tissues of some patients with chronic kidney disease (CKD) were determined using rabbit anti-AOPP-RSA.Results:Titers of the antibodies were 10-6.Purified antibodies reacted specifically with oxidized albumin from different genus,but could not react with normal albumin and glycosylated albumin.The high level of AOPP in plasma from CKD patients was confirmed by Western blot.The antibodies could be used to immunostain AOPP deposition in different regions of kidney tissues from both CKD patients and CKD rat models.Conclusion:We successfully generate rabbit anti-AOPP polyclonal antibodies with high titers and striking specificity.The presence of plasma AOPP and localizations of AOPP in kidney tissues of CKD patients can be demonstrated using the antibodies.The development of anti-AOPP polyclonal antibodies may provide a new tool to explore the pathogensis of AOPP and assess exactly the relationship between AOPP and relative diseases.

AIM: To investigate the effect of advanced glycosylation end products(AGEs) modified protein on morphological changes of actin cytoskeleton in primary endothelial cells and the role of MAPK signaling pathways in this pathological procedure.METHODS: Human umbilical vein endothelial cells(HUVECs) were incubated with AGEs modified human serum albumin(AGE-HSA) at concentrations of 12.5,25,50,and 100 mg/L,respectively,for 2,4,8,12 and 24 hours.The cells were treated with different chemical compounds of inhibitors,or adenoviral deliver approach(either dominant positive or negative adenoviral constructs) of MAP kinases to specifically block or activate certain signal transduction pathways under above situations.As control,HSA of the same concentration was administered to cells at the same time.The treated cells were incubated with FITC-phalloidin to stain F-actin.RESULTS: F-actin in HUVECs was rearranged greatly by AGEs in a concentration and time-dependent manners.The unmodified HSA did not influence Factin distributions.The AGEs-induced changes were blocked by pretreatment with SB203580,PD98059 for 30 min,or pre-infection with recombinant virus of dominant negative form of MKK 6b [MKK6b(A)],MEK1 [MEK1(A)] for 24 h,while SP600125 and dominant negative form of MKK7 [MKK7(A)] failed to inhibit the effects of AGEs.Furthermore,the infection of recombinant virus of constitutive active form of MKK6b [MKK6b(E)] or MEK1 [MEK1(E)] could also induced re-arrangement of F-actin,respectively,while the effect elicited by [MKK6b(E)] was abolished by co-infection with recombinant adeno-virus of dominant negative p38?. CONCLUSION: AGEs-induced morphological changes of F-actin in endothelial cells are mediated by p38-and ERK MAPK signal pathways.

Objective: To study the effect of high-dose methylprednisolone intravenous pulse therapy on immunoglobulins and CD series in patients with active moderate-to-severe Graves′ orbitopathy. Methods: Twenty-seven patients with active moderate-to-severe Graves′ orbitopathy were enrolled in this study. All the patients received iv methylprednisolone pulse therapy for 12 weeks according to the 2016 European Thyroid Association/European Group on Graves′Orbitopathy(EUGOGO) Guidelines. Serum thyroidal autoantibodies, such as thyroid-stimulating hormone receptor antibody (TRAb), anti-thyroperoxidase antibody (TPOAb), and serum immunoglobulins, such as IgG, IgE, IgA, IgM were evaluated at the baseline, at the end of 4th and 12th week. Percentages of CD3⁺ T cells, CD4⁺ T cells, CD8⁺ T cells and CD19⁺ B cells, CD16⁺ or CD56⁺ NK cells were also evaluated at each time point. Results: TRAb, TPOA and IgE, IgG, IgA were significantly decreased both after 4th week and after 12th week (all P<0.05). Percentages of CD3⁺ T cells, CD4⁺ T cells and CD4⁺ /CD8⁺ ratio were gradually decreased after treatment. However, change of CD8⁺ T cells was not significant (P>0.05). Conclusion High-dose methylprednisolone may exert an immunosuppressive effect not only by modifying humoral and cellular immune functions, but also by decreasing serum immunoglobulins.

OBJECTIVE: To explore molecular etiology and clinical characteristics of two pedigrees affected with hereditary factor VII(FVII) deficiency. METHODS: The nine exons and flanking sequences of the F7 gene of the probands were amplified by PCR. The amplicons were analyzed by direct sequencing. Suspected mutations were subjected to SWISS-MODEL modeling and analysis of protein structure change by Pymol software and conservation of amino acids across various species. RESULTS: For proband of pedigree 1, the prothrombin time (PT), FVII activity (FVII:C) and FVII antigen (FVII:Ag) were 36.3 s, 3%, 53.56%, respectively. Sequencing revealed a compound heterozygous variants of c.80_81delCT and c.1371G>T(p.Arg439Ser). His son carried a heterozygous c.1371G>T (p.Arg439Ser) variant. For proband of pedigree 2, the PT, FVII:C and FVII:Ag were 22.3 s, 4%, 1.58%, respectively. Sequencing has revealed a compound heterozygous c.278G>T(p.Arg75Met) missense variant in exon 3 and c.1278T>G (p.His408Gln) in exon 9 of the F7 gene. His mother and son both carried a heterozygous c.278G>T(p.Arg75Met) variant. Three-dimensional simulation and homology analysis revealed that the p.Arg439Ser and p.Arg75Met can respectively alter part of hydrogen bonds and two highly conserved amino acids. CONCLUSION Two novel heterozygous missense variants of the F7 gene [c.1371G>T(p.Arg439Ser) and c.278G>T(p.Arg75Met)] probably account for the decrease of factor VII in the two pedigrees.
Asian Continental Ancestry Group ; Factor VII ; Factor VII Deficiency ; Genotype ; Heterozygote ; Humans ; Mutation ; Pedigree

Objective:To analyze the clinicopathological features in diabetic kidney disease (DKD) and non-diabetic kidney disease (NDKD) patients, and provide reference for patients who will receive renal biopsy with diabetes mellitus complicated with chronic kidney disease.Methods:The patients with type 2 diabetes mellitus complicated with chronic kidney disease who underwent renal biopsy were collected through the database at the Nanfang Hospital of Southern Medical University from February 2002 to June 2018. According to the results of renal biopsy, they were divided into DKD group and NDKD group (including DKD+NDKD). The clinical manifestations and pathological types were compared between the two groups.Results:A total of 507 patients were eventually included in the study. There were 114 cases (22.5%) with DKD and 393 cases (77.5%) with NDKD. Pathologically, the most common pathological types of NDKD were membranous nephropathy (30.0%) and IgA nephropathy (19.1%). Among NDKD patients, 5.6% patients had DKD combing with NDKD. In term of the clinical manifestations, DKD patients had a longer history of diabetes (>1 year, 76.3% vs 36.1%, P<0.001), higher quantity of urinary protein [3.69(1.70, 6.74) g/24 h vs 2.21(0.91, 4.97) g/24 h, P<0.001], higher serum creatinine [117.5(85.8, 194.5) μmol/L vs 89.0(68.0, 143.8) μmol/L, P<0.001] than NDKD patients. But the hemoglobin [(105.07±20.85) g/L vs (124.41±25.02) g/L, P=0.002] and cholesterol [(5.69±1.87) mmol/L vs (6.43±2.75) mmol/L, P=0.001] in DKD patients were lower than those in NDKD patients. Logistic regression analysis showed that diabetes mellitus history ( OR=4.162, 95% CI 1.717-10.098, P=0.002) , higer systolic pressure (every 1 mmHg, OR=1.028, 95% CI 1.011-1.045, P=0.001) , history of antihypertensive medication ( OR=3.141, 95% CI 1.496-6.591, P=0.002), diabetic retinopathy ( OR=5.561, 95% CI 2.361-13.100, P<0.001) and higher glycated hemoglobin level (every 1%, OR=1.680, 95% CI 1.333-2.118, P<0.001) were related factors of DKD, while hematuria ( OR=2.781, 95% CI 1.334-5.798, P=0.006) and higher hemoglobin level (every 1 g/L, OR=1.022, 95% CI 1.008-1.037, P=0.002) were related factors of NDKD. Conclusions:There are differences in clinical manifestations and pathological types between DKD and NDKD. The history of diabetes, antihypertensive medication, fundus examination, higher of proteinuria and glycosylated hemoglobin may predict DKD, while hematuria and higher level of hemoglobin may have certain guiding significance for the diagnosis of NDKD. The indication of renal biopsy in patients with diabetes mellitus complicated with chronic kidney disease should include comprehensive clinical manifestations.

Gaining a better understanding of autoprotection against drug-induced liver injury(DILI)may provide new strategies for its prevention and therapy.However,little is known about the underlying mechanisms of this phenomenon.We used single-cell RNA sequencing to characterize the dynamics and functions of hepatic non-parenchymal cells(NPCs)in autoprotection against DILI,using acetaminophen(APAP)as a model drug.Autoprotection was modeled through pretreatment with a mildly hepatotoxic dose of APAP in mice,followed by a higher dose in a secondary challenge.NPC subsets and dynamic changes were identified in the APAP(hepatotoxicity-sensitive)and APAP-resistant(hepatotoxicity-resistant)groups.A chemokine(C-C motif)ligand 2+endothelial cell subset almost disappeared in the APAP-resistant group,and an R-spondin 3+endothelial cell subset promoted hepatocyte proliferation and played an important role in APAP autoprotection.Moreover,the dendritic cell subset DC-3 may protect the liver from APAP hepatotoxicity by inducing low reactivity and suppressing the autoimmune response and occurrence of inflammation.DC-3 cells also promoted angiogenesis through crosstalk with endothelial cells via vascular endothelial growth factor-associated ligand-receptor pairs and facilitated liver tissue repair in the APAP-resistant group.In addition,the natural killer cell subsets NK-3 and NK-4 and the Sca-1-CD62L+natural killer T cell subset may promote autoprotection through interferon-y-dependent pathways.Furthermore,macrophage and neutrophil subpopulations with anti-inflammatory phenotypes promoted tolerance to APAP hepatotoxicity.Overall,this study reveals the dynamics of NPCs in the resistance to APAP hepatotoxicity and provides novel insights into the mechanism of autoprotection against DILI at a high resolution.

【Objective】 To investigate the effect of spleen on hepatic macrophages mediated activation of hepatic stellate cells (HSCs) in mice with liver fibrosis. 【Methods】 Eighteen male C57BL/6 mice were randomly divided into three groups. Mice in Group A and Group B were injected intraperitoneally with CCl4 to establish liver fibrosis mouse model, while those in Group C were injected with corn oil as normal control. Four weeks later, mice with liver fibrosis received splenectomy (Spx) or sham operation (Sham), respectively. After continuous injection for 2 weeks, liver homogenates (L-Homo) were prepared and liver cells were isolated from the three groups. Expressions of IL-1β, IL-13, TGF-β, TNF-α, PDGF-β and VEGF in the liver homogenates of the three groups were detected by Luminex multifactor analysis. The expressions of these cytokines in liver macrophages (L-Mφ) and other non-parenchymal cells of Sham and Spx mice were analyzed by Real-time quantitative PCR (RT-qPCR) and flow cytometry. Macrophage cell line RAW264.7 or bone marrow-derived macrophages (BMDMs) were treated with liver homogenates from the Sham and Spx groups. Then the differently treated RAW264.7 cells were analyzed for mRNA expressions of cytokines and glutamine metabolism-related molecules by RT-qPCR, or transwell co-cultured with hepatic stellate cell line JS1. After co-culture, the survival and extracellular matrix expression of JS1 cells were analyzed. For comparison, Student’s t test (between two groups) or one-way analysis of variance (among multiple groups) were used. 【Results】 Compared with normal control group, the concentrations of IL-1β, IL-13, TGF-β and TNF-α in the L-Homo of model group were significantly increased and showed higher levels in Sham group than in Spx group. Moreover, the hepatic macrophages were indicated as the major source of these cytokines. Consistently, macrophages treated with liver homogenate of Sham mice had increased expressions of IL-1β, TGF-β and TNF-α and glutaminase (GLS). After co-culture with macrophages treated with liver homogenate of Sham group rather than Spx group, JS1 expressed higher expressions of α-SMA and collagens. 【Conclusion】 The spleen is involved in regulating the secretion of cytokines by hepatic macrophages and enhancing their ability to activate hepatic stellate cells.