Latest research and prospect of CD47 in kidney transplantation / 器官移植

CD47 is a transmembrane protein widely expressed on cell surface, which is considered as a key molecule for immune escape. With an increasing number of related studies, the role of CD47 and its ligands in immunomodulatory effects has been gradually understood. Recent studies have investigated the role of CD47 in ischemia-reperfusion injury of allogenetic kidney transplantation, rejection and xenotransplantation. Nevertheless, the specific role and the key mechanism remain elusive. In this article, the structure and function of CD47, common CD47 ligands, the relationship between CD47 and kidney transplantation, and the application of CD47 in kidney transplantation were reviewed, the latest research progress of CD47 in kidney transplantation was summarized, and the limitations of current research and subsequent research direction were analyzed, aiming to provide reference for subsequent application of CD47 in allogeneic and kidney xenotransplantation.

Clinical Significance of Thrombospondin Type 1 Domain-Containing 7A and Neural Epidermal Growth Factor-Like 1 Protein in M-Type Phospholipase A2 Receptor-Negative Membranous Nephropathy / 中国医学科学院学报

Objective To investigate the clinical significance of thrombospondin type 1 domain-containing 7A (THSD7A) and neural epidermal growth factor-like 1 protein (NELL1) in phospholipase A2 receptor (PLA2R)-negative membranous nephropathy (MN). Methods A total of 116 PLA2R-negative MN patients treated in Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University from 2014 to 2021 were enrolled in this study.Immunohistochemistry was employed to detect THSD7A and NELL1 in the renal tissue.The pathological characteristics,treatment,and prognosis were compared between positive and negative groups. Results The 116 PLA2R-negative MN patients included 23 THSD7A-positive patients and 9 NELL1-positive patients.One patient was tested positive for both proteins.The THSD7A-positive group showed higher positive rate of IgG4 (P=0.010),more obvious glomerular basement membrane (GBM) thickening (P=0.034),and higher proportion of stage Ⅱ MN and lower proportion of stage I MN (P=0.002) than the THSD7A-negative group.The NELL1-positive group had lower positive rates of C1q and IgG2 (P=0.029,P=0.001),less obvious GBM thickening (P<0.001),more extensive inflammatory cell infiltration (P=0.033),lower proportion of deposits on multi-locations (P=0.001),and lower proportion of atypical MN (P=0.010) than the NELL1-negative group.One patient with THSD7A-positive MN was diagnosed with colon cancer,while none of the NELL1-positive patients had malignancy.Survival analysis suggested that THSD7A-positive MN had worse composite remission (either complete remission or partial remission) of nephrotic syndrome than the negative group (P=0.016),whereas NELL1-positive MN exhibited better composite remission of nephrotic syndrome than the negative group (P=0.015).The MN patients only positive for NELL1 showed better composite remission of nephrotic syndrome than the MN patients only positive for THSD7A (P<0.001). Conclusions THSD7A- and NELL1-positive MN is more likely to be primary MN,and there is no significant malignancy indication.However,it might have a predictive value for the prognosis of MN.

Predictive value of serum thrombospondin-1 and transforming growth factor-β1 levels in preterm infants with bronchopulmonary dysplasia / 中国小儿急救医学

Objective:To study the early predictive values of serum thrombospondin-1(TSP-1)and transforming growth factor-β1(TGF-β1) for bronchopulmonary dysplasia(BPD)in preterm infants.Methods:From September 2020 to April 2022, preterm infants with gestational age<32 weeks and ≥28 weeks as well as birth weight<1 500 g admitted to neonatal intensive care unit within 2 hours after birth were enrolled in the study.The dynamic changes of serum TSP-1 and TGF-β1 levels in preterm infants were observed on 1st, 7th, 14th, and 28th day after birth.Preterm infants were divided into BPD group and non-BPD group according to the diagnostic criteria of BPD.Receiver operating characteristic(ROC) curve and area under curve(AUC)was used to analyze the predictive value of serum TSP-1 and TGF-β1 for preterm infants with BPD.Results:According to the diagnostic criteria of BPD, 38 cases were in the BPD group and 52 cases in the non-BPD group.There was no significant difference in gestational age, birth weight and gender between the two groups( P>0.05). The levels of TSP-1 and TGF-β1 in the serum of BPD group were gradually increased, which were significantly higher than those of non-BPD group on the 1st, 7th, 14th, and 28th day( P<0.001). ROC results showed that AUC of TSP-1, TGF-β1 and their combination for predicting BPD were 0.889(95% CI 0.819~0.959), 0.826(95% CI 0.743~0.910), and 0.923(95% CI 0.870~0.976), respectively.The sensitivity were 86.80%, 86.70%, 89.50%, and the specificity were 86.50%, 73.10%, 80.80%, respectively.Cutoff values of TSP-1 and TGF-β1 for predicting BPD were 44.50 μg/L and 6.13 μg/L, respectively. Conclusion:Combined detection of serum TSP-1 and TGF-β1 on the first day after birth has an early predictive value for BPD in preterm infants.

Improving cancer immunotherapy via co-delivering checkpoint blockade and thrombospondin-1 downregulator

The use of checkpoint-blockade antibodies is still restricted in several malignancies due to the modest efficacy, despite considerable success in anti-tumor immunotherapy. The poor response of cancer cells to immune destruction is an essential contributor to the failure of checkpoint therapy. We hypothesized that combining checkpoint therapy with natural-product chemosensitizer could enhance immune response. Herein, a targeted diterpenoid derivative was integrated with the checkpoint blockade (anti-CTLA-4) to improve immunotherapy using thermosensitive liposomes as carriers. In vivo, the liposomes enabled the co-delivery of the two drug payloads into the tumor. Consequently, the regulatory T cell proliferation was restrained, the cytotoxic T cell infiltration was enhanced, and the profound immunotherapeutic effect was achieved. In addition, the immunotherapeutic effect of another clinically used checkpoint antibody, anti-PD-1, also benefited from the diterpenoid derivative. Of note, our mechanism study revealed that the targeted diterpenoid derivative increased the sensitivity of cancer cells to immune attack via THBS1 downregulation and the resultant destruction of THBS1-CD47 interaction. Collectively, co-delivering THBS1 inhibitor and checkpoint blockade is promising to boost cancer immunotherapy. We first time discovered that THBS1 suppression could strengthen checkpoint therapy.

Effect of Xiaoxuming Decoction on activation of astrocytes in acute cerebral ischemia/reperfusion injury / 中国中药杂志

This study investigated the effect of Xiaoxuming Decoction(XXMD) on the activation of astrocytes after cerebral ischemia/reperfusion(I/R) injury. The model of cerebral IR injury was established using the middle cerebral artery occlusion method. Fluorocitrate(FC), an inhibitor of astrocyte activation, was applied to inhibit astrocyte activation. Rats were randomly divided into a sham group, a model group, a XXMD group, a XXMD+FC group, and a XXMD+Vehicle group. Neurobehavioral changes at 24 hours after cerebral IR injury, cerebral infarction, histopathological changes observed through HE staining, submicroscopic structure of astrocytes observed through transmission electron microscopy, fluorescence intensity of glial fibrillary acidic protein(GFAP) and thrombospondin 1(TSP1) measured through immunofluorescence, and expression of GFAP and TSP1 in brain tissue measured through Western blot were evaluated in rats from each group. The experimental results showed that neurobehavioral scores and cerebral infarct area significantly increased in the model group. The XXMD group, the XXMD+FC group, and the XXMD+Vehicle group all alleviated neurobehavioral changes in rats. The pathological changes in the brain were evident in the model group, while the XXMD group, the XXMD+FC group, and the XXMD+Vehicle group exhibited milder cerebral IR injury in rats. The submicroscopic structure of astrocytes in the model group showed significant swelling, whereas the XXMD group, the XXMD+FC group, and XXMD+Vehicle group protected the submicroscopic structure of astrocytes. The fluorescence intensity and protein expression of GFAP and TSP1 increased in the model group compared with those in the sham group. However, the XXMD group, the XXMD+FC group, and XXMD+Vehicle group all down-regulated the expression of GFAP and TSP1. The combination of XXMD and FC showed a more pronounced effect. These results indicate that XXMD can improve cerebral IR injury, possibly by inhibiting astrocyte activation and down-regulating the expression of GFAP and TSP1.

Expression of CD47 and its ligands in pregnant mice infected with Toxoplasma gondii during pregnancy / 中国血吸虫病防治杂志

Objective To investigate the dynamic expression of cluster of differentiation 47 (CD47) and its ligands signaling regulatory protein α (SIRPα) and thrombospondin-1 (TSP-1) in mice infected with Toxoplasma gondii in the second and third trimesters.. Methods C57BL/6J mice (6 to 8 weeks old) were used for modeling T. gondii infection in the first trimester, and the pregnant mice were randomly divided into the normal control and infection groups, of 10 mice in each group. Pregnant mice in the infection group were intraperitoneally injected with 150 T. gondii tachyzoites on gestational day (Gd) 6.5, while pregnant mice in the normal control group were intraperitoneally injected with the same volume of physiological saline at the same time. The uterine and placental specimens were collected from all pregnant mice on Gd12.5 and Gd18.5, and the pregnant outcomes were recorded. The pathological damages of mouse uterine and placental specimens were observed using hematoxylin-eosin (HE) staining on Gd12.5 and Gd18.5. The relative expression of CD47, SIRPα, TSP-1, surface antigen 1 (SAG1), interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4 and IL-13 mRNA was quantified in mouse uterine and placental specimens using real-time fluorescence quantitative PCR (qPCR) assay, and the CD47, SIRPα, TSP-1 expression was determined in mouse uterine and placental specimens using immunohistochemical staining. Results As compared with those in the normal control group, the pregnant mice in the infection group showed back arching, bristling, trembling and listlessness during pregnancy, and several mice presented virginal bleeding and abortion. Pathological examinations showed inflammatory cell infiltration, congestion and necrosis in uterine and placental specimens of pregnant mice in the infection group, a higher abortion rate of pregnant mice was seen in the infection group than in the normal control group on Gd12.5 (χ2 = 20.405, P < 0.001) and Gd18.5 (χ2 = 28.644, P < 0.001). qPCR assay showed significant differences in the expression of CD47, SIRPα, TSP-1, SAG1, INF-γ, IL-2, IL-4 and IL-13 genes in mouse placental specimens between the normal control and infection groups on Gd12.5 and Gd18.5 [F′ (F) = 37.511, 29.337, 97.343, 53.755, 67.188, 21.145, 8.658 and 13.930, all P values < 0.001]. Higher CD47, SIRPα and TSP-1 gene expression was quantified in mouse placental specimens in the infection group than in the normal control group on Gd12.5 (all P values < 0.01), and lower CD47, SIRPα and TSP-1 gene expression was quantified in the infection group than in the normal control group on Gd18.5 (all P values < 0.001), while higher SAG1 gene expression was detected in placental specimens of pregnant mice in the infection group than in the normal control group on Gd12.5 and Gd18.5 (both P values < 0.01). In addition, higher INF-γ and IL-2 expression and lower IL-4 and IL-13 expression was detected in mouse placental specimens in the infection group than in the normal control group on Gd12.5 and Gd18.5 (all P values < 0.001), and there were significant differences in the CD47, SIRPα, TSP-1, SAG1, INF-γ, IL-2, IL-4 and IL-13 gene expression in uterine specimens of pregnant mice between the normal control and infection groups on Gd12.5 and Gd18.5 [H(F′ and F) = 14.951, 25.977, 18.711, 48.595, 39.318, 14.248 and 15.468, all P values < 0.01], and higher CD47 and TSP-1 expression was detected in mouse uterine specimens in the infection group than in the control group on Gd12.5 and Gd18.5 (all P values < 0.01); however, no significant difference was found in the SIRPα expression (P > 0.05). Higher SAG1 expression was detected in uterine specimens of pregnant mice in the infection group than in the normal control group on Gd12.5 and Gd18.5 (both P values < 0.01), and higher INF-γ and IL-2 gene expression and lower IL-4 and IL-13 gene expression was found in the placental specimens of pregnant mice in the infection group than in the normal control group on Gd12.5 and Gd18.5 (all P values < 0.001). Spearman correlation analysis showed that the CD47 gene expression correlated positively with IFN-γ (rs = 0.735, P < 0.05) and IL-2 (rs = 0.655, P < 0.05) and negatively with IL-4 (rs = −0.689, P < 0.05) and IL-13 expression (rs = −0.795, P < 0.05) in the placental specimens of pregnant mice in the infection group on Gd12.5, and the CD47 gene expression correlated negatively with IFN-γ (rs = −0.745, P < 0.05) and IL-2 expression (rs = −0.816, P < 0.05) and positively with IL-4 (rs = 0.704, P < 0.05) and IL-13 (rs = 0.802, P < 0.05) in the placental specimens of pregnant mice in the infection group on Gd18.5. Immunohistochemical staining showed mild CD47, SIRPα and TSP-1 expression in uterine and placental specimens of pregnant mice in the normal control group on Gd12.5 and Gd18.5, strong CD47, SIRPα and TSP-1 expression in the placental specimens of pregnant mice in the infection group on Gd12.5 and strong CD47 and TSP-1 expression in the uterine specimens of pregnant mice in the infection group on Gd12.5. Conclusions T. gondii infection in the first trimester may cause abnormal expression of CD47 and its ligands SIRPα and TSP-1 in the maternal-fetal interface of pregnant mice in the second and third trimesters, which may be associated with the immune escape of T. gondii at the maternal-fetal interface.

THBS4 Promotes the Metastasis of Hepatocellular Carcinoma Cells by Inducing M2 Polarization of Tumor-associated Macrophages / 中国生物化学与分子生物学报

Thrombospondin 4 (THBS4), a member of the THBS family, is a protein secreted by the extracellular matrix and is involved in regulating various physiological processes, such as cell proliferation, adhesion and angiogenesis. Recent studies have shown that the inflammation stimulates THBS4 production and induces the adhesion and accumulation of macrophages. Our previous study confirmed that THBS4 acts as an oncogene in hepatocellular carcinoma (HCC), the effect of THBS4 on the immune microenvironment of HCC remains unclear. This study aims to analyze the role of THBS4 in promoting the metastasis of HCC cells by inducing M2-type polarization of tumor-associated macrophages. We simulate the tumor microenvironment through HCC conditioned medium (HCM) and found that the expression of THBS4 in macrophages increased in a time-dependent manner under the action of HCM (P<0.05); THBS4 knockdown promotes the expression of M1 macrophages markers IL-1β and CD86 (P<0.01), while the expression of M2-type markers IL-10 and CD206 were decreased (P<0.01). Transwell co-culture assay was used to further detect the effect of THBS4-induced M2-type macrophages on HCC metastasis. Results from co-culture of THBS4-downregulated M2 macrophages with HepG2 cells showed that THBS4-downregulated M2-TAMs significantly inhibited the invasion and migration ability of HepG2 cells (all P < 0.01). In conclusion, the tumor microenvironment promotes the expression of THBS4 in macrophages, and THBS4 may promote the invasion and metastasis of HCC cells by inducing M2-type polarization of macrophages. This study provides some new experimental basis for exploring the establishment of THBS4-induced HCC immune microenvironment.

The Changes of Peripheral Blood FⅫa-AT, TSP-1, LA Ratio in Patients with Systemic Lupus Erythematosus and the Clinical Value of Combined Diagnosis of Thrombotic Events / 中国实验血液学杂志

OBJECTIVE@#To explore the changes of Ⅻ antithrombin (FⅫa-AT), thrombospondin-1 (TSP-1), and lupus anticoagulant (LA) ratio in the peripheral blood factor of patients with systemic lupus erythematosus (SLE) and the clinical value of combined diagnosis of thrombotic events.@*METHODS@#A total of 133 SLE patients treated in Xingtai People's Hospital were selected and divided into simple SLE group (105 cases) and SLE complicated with thrombosis group (28 cases) according to whether thrombotic events occurred, and 102 cases of healthy people in the same period were selected as control. The clinical data of the 3 groups, the level of peripheral blood FⅫa-AT, TSP-1, and LA ratio were compared, the relationship between each peripheral blood index and SLE disease activity index (SLEDAI) score were analyzed. The influencing factors of thrombotic events in SLE patients were analyzed, and the value of each peripheral blood index in the diagnosis of SLE complicated with thrombotic events were evaluated.@*RESULTS@#The proportion of the patients with age ≥60 year, hypertension, and smoking history in SLE complicated with thrombosis group was higher than those in simple SLE group and control group (P＜0.05). The SLEDAI score, peripheral blood FⅫa-AT, TSP-1, LA ratio levels of the patients in SLE complicated with thrombosis group were significantly higher than those in simple SLE group and control group, and the simple SLE group was significantly higher than the control group (P＜0.05). FⅫa-AT, TSP-1, LA ratio in peripheral blood of SLE patients were positively correlated with SLEDAI score (r=0.663, 0.578 and 0.625). Age, blood pressure, smoking history, peripheral blood FⅫa-AT, TSP-1, LA ratio were the important influencing factors of thrombotic events in SLE patients (P＜0.05). The AUC diagnosed by the FⅫa-AT, TSP-1, and LA ratio in peripheral blood was 0.881, the 95% CI was 0.813-0.931, the sensitivity was 82.14%, and the specificity was 91.43%, which was superior to each index alone (P＜0.05).@*CONCLUSION@#Peripheral blood FⅫa-AT, TSP-1, LA ratio level changes in SLE patients are significantly related to disease activity, and the combined diagnosis of thrombotic events is more reliable.

Effects of plasma lipopolysaccharide changes on platelet release of vascular endothelial growth factor and thromobospondin-1 in patients with cirrhotic portal hypertension after TIPS procedure / 中华肝脏病杂志

Objective: To investigate the effects of plasma lipopolysaccharide (LPS) concentration changes on platelet release of vascular endothelial growth factor (VEGF) and thrombospondin (TSP)-1 in patients with decompensated cirrhotic portal hypertension after transjugular intrahepatic portosystemic shunt (TIPS) procedure. Methods: 169 cases with cirrhotic portal hypertension were enrolled, of which 81 cases received TIPS treatment. LPS, VEGF, and TSP-1 concentrations with different Child-Pugh class in peripheral blood plasma of patients were measured. After pre-incubation of normal human platelets with different concentrations of LPS and stimulated by collagen in vitro, platelet PAC-1 expression rate, VEGF, and TSP-1 concentrations were detected. PAC-1 expression rate and the concentrations of LPS, VEGF and TSP-1 in peripheral blood plasma of patients before and after TIPS procedure were detected. The relationship between plasma LPS, VEGF and TSP-1 concentrations and Child-Pugh score changes in patients after TIPS procedure was analyzed. Statistical analysis was performed by t-test, one-way ANOVA or Pearson's rho according to different data. Results: Plasma LPS and TSP-1 concentrations were significantly higher in Child-Pugh class C patients than class A and B, but the concentration of plasma VEGF was significantly lower than class A and B (P < 0.01). In vitro experiments showed that concentration of LPS, TSP-1, and platelet PAC-1 expression rate was higher in the supernatant, but the difference in the concentration of VEGF in the supernatant was not statistically significant. Portal vein pressure and platelet activation were significantly decreased (P < 0.01) in patients after TIPS procedure. Portal venous pressure, platelet activation, plasma LPS, and TSP-1 levels were significantly decreased continuously, while VEGF levels were significantly increased continuously after TIPS procedure. Plasma LPS concentration was positively correlated with TSP-1 concentration (r = 0.506, P < 0.001), and negatively correlated with VEGF concentration (r = -0.167, P = 0.010). Child-Pugh score change range was negatively correlated with change range of plasma VEGF concentration (r = -0.297, P = 0.016), and positively correlated with change range of plasma TSP-1 concentration (r = 0.145, P = 0.031) after TIPS. Conclusion: Portal venous pressure gradient, plasma LPS concentration and corresponding platelet activation was decreased in cirrhotic portal hypertension after TIPS procedure, and with TSP-1 reduction and VEGF elevation it is possible to reduce the liver function injury caused by portal venous shunt.

A case of acral fibrochondromyxoid tumor / 中华皮肤科杂志

A 44-year-old male patient presented with a subcutaneous nodule in the left little finger for 3 years. Skin examination showed a subcutaneous nodule with rubber-like hardness but no tenderness on palpation, measuring 0.4 cm × 0.4 cm in size at the dorsal distal aspect of the left little finger, and the movement of the distal interphalangeal joint was unrestricted. Postoperative histopathological examination revealed that the tumor contained abundant stroma consisting of variable fiberous, chondroid and myxoid materials; tumor cells were oval to short spindle-shaped with inconspicuous nucleoli but no mitosis; cells were arranged haphazardly or in small clusters. Immunohistochemical study showed positive staining for vimentin, CD34 and transcription factors ERG and SOX9, but negative staining for S100, P63, broad-spectrum cytokeratin AE1/AE3, epithelial membrane antigen, smooth muscle actin and desmin in tumor cells, and the Ki67 labeling index was below 1%. Finally, the patient was diagnosed with acral fibrochondromyxoid tumor.

Study on the correlation of integr in associated protein and disease activity and bone destruction in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective:To analyze the serum levels of integrin-associated proteins (CD47) in patients with rheumatoid arthritis (RA), and to explore its association with disease activity and bone destruction in RA.Methods:Serum and clinical data were collected from 65 RA patients and 25 healthy subjects. RA patients were grouped into low, moderate, and high bone erosion groups according to 7-joint ultrasonography score (US7). The levels of serum CD47, thrombospondin-1 (TSP-1) and receptor activator of nuclear factor-κB ligand (RANKL) were measured by enzyme-linked immunosorbnent assay (ELISA) in patients with RA and healthy subjects. The statistical analysis was carried out with independent t-test, analysis of variance, nonparametric rank sum test, pearson or Spearman correlation and logistic regression. Results:① The Serum levels of CD47, TSP-1, and RANKL were higher in the RA group than in the healthy controls ( P<0.01). ② In RA patients, serum CD47 level was positively correlated with disease course ( r=0.301, P<0.05), C-reactionprotein (CRP)( r=0.316, P<0.05), number of tender joints (TJC) ( r=0.254, P<0.05), number of swollen joints (SJC) ( r=0.316, P<0.05), disease activity score in 28 joints (DAS28) ( r=0.255, P<0.05), RANKL ( r=0.252, P<0.05) and TSP-1 ( r=0.260, P<0.05). Serum TSP-1 level was positively correlated with CRP ( r=0.299, P<0.05), TJC ( r=0.335, P<0.01), DAS28 ( r=0.315, P<0.05), RANKL ( r=0.305, P<0.05). ③ The disease course [ OR(95% CI)=1.048(1.033, 1.017)] and TSP-1 [ OR(95% CI)=1.013(1.000, 1.026)] were independently relevant factors affecting bone destruction. Conclusion:CD47 levels is significantly higher in RA patients than in healthy controls, and is associated with disease activity and bone destruction. CD47 may be involved in the bone destruction process of RA by acting on TSP-1.

Correlation between serum NLR, TSP-1, miR-210 and systemic lupus erythematosus disease activity index and their combined prognostic value / 中国医师杂志

Objective:To investigate the correlation between serum neutrophil to lymphocyte ratio (NLR), thrombospondin 1 (TSP-1), miR-210 and systemic lupus erythematosus disease activity index (SLEDAI) and the prognostic value of their combination.Methods:The medical records of 126 patients with systemic lupus erythematosus (SLE) in Haikou People′s Hospital (Haikou Hospital Affiliated to Xiangya Medical College of Central South University) from February 2018 to February 2020 were retrospectively analyzed. According to the recurrence of prognosis 6 months after treatment, they were divided into recurrence group ( n=23) and non recurrence group ( n=103). The general data, serum NLR, TSP-1, miR-210 levels and SLEDAI score before and after treatment of the two groups were compared. The relationship between the levels of serum indicators before and after treatment, SLEDAI score, prognosis and recurrence of SLE patients were analyzed, and the efficacy of single and combined serum indicators in predicting prognosis was explored. Results:The levels of serum NLR, TSP-1, miR-210 and SLEDAI score in the recurrence group were higher than those in the non recurrence group before and after treatment (all P<0.05); After treatment, the levels of serum NLR, TSP-1, miR-210 and SLEDAI score in the two groups were lower than those before treatment (all P<0.05). Pearson correlation analysis showed that the levels of serum NLR, TSP-1 and miR-210 in SLE patients were positively correlated with SLEDAI scores (all P<0.05); Cox regression analysis showed that after adjusting other factors such as complement C3, complement C4 levels and SLEDAI scores before and after treatment, serum NLR, TSP-1 and miR-210 before and after treatment were still significantly correlated with the risk of recurrence in SLE patients (all P<0.05); The receiver operating characteristic (ROC) curve showed that the area under curve (AUC) of serum NLR, TSP-1 and miR-210 combined to predict recurrence was 0.907 (95% CI: 0.842-0.951), the sensitivity was 86.96%, and the specificity was 83.50%, which was significantly higher than that of each index alone. Conclusions:Serum NLR, TSP-1, miR-210 levels in SLE patients are positively correlated with SLEDAI scores, and the combined detection of these indicators has a high predictive value for prognosis and recurrence, which can provide references for the diagnosis and treatment of SLE.

Effect of Modifide Wenjingtang on JAK2/STAT3 Signal Pathway and Downstream Factors Protein Expression in Endometriosis Rats / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect and mechanism of modified Wenjingtang on endometriosis （EM） rats with kidney deficiency and blood stasis. Method:The 10 from 105 SPF female healthy SD rats were randomly selected as the blank group. The rest constructed the rat model of kidney deficiency and blood stasis by compound factorial method. After the model was successfully established， 10 rats were randomly selected as the sham operation group， with only laparotomy and no intima suture， and the remaining rats were established with EM kidney deficiency and blood stasis type by autologous intimal transplantation. Fifty rats which were randomly selected from 56 successful rats were treated with the modified Wenjingtang （5，10，20 g·kg^-1） and danazol group（63 mg·kg^-1）， 1 time daily ， for 4 weeks. The endometrial tissues of each group were stained with hematoxylin eosin （HE） to observe the histopathology. The levels of inflammatory factors interleukin-10 （IL-10） and interleukin-17 （IL-17） in serum supernatant were detected by enzyme linked immunosorbent assay （ELISA）. Measuring the length（D₁），width （D₂） and height （D₃） of the heterotopic foci in each group before and after treatment. Then calculating the volume of them. The expression of tyrosine kinase 2（JAK2），transcription factor 3 （STAT3），phosphorylation transcription factor 3 （p-STAT3）， vascular endothelial growth factor （VEGF）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） and thrombospondin-1 （TSP-1） were detected by immunohistochemistry （IHC）. The expression of VEGF，TNF-<italic>α</italic> and TSP-1 was detected by Western blot. Result:Microscopic pathological observation showed that the endometrial glandular cells of the blank group were arranged in order， and the glandular and stromal cells grew well， compared with the blank group， the endometrial structure of the model group was complete， showing a cavity like or annular closed structure， with cyst formation， and the epithelium was cubic or columnar epithelium， most of the epithelial cells had secretion， the stroma was dense， and the matrix showed a little fibrosis There were a few glands and inflammatory cell infiltration. Compared with the blank group， the content of IL-10 in serum of model group was significantly decreased （<italic>P</italic><0.01）， and the content of IL-17 was significantly increased （<italic>P</italic><0.01）， the protein expression of JAK2， STAT3，p-STAT3， VEGF， TNF-<italic>α</italic> in endometrial tissue of model group was significantly increased （<italic>P</italic><0.05）， and the expression of TSP-1 protein was significantly decreased （<italic>P</italic><0.05）. Compared with the model group， the serum IL-10 content of rats in modified Wenjingtang treatment group increased significantly （<italic>P</italic><0.01）， the IL-17 content decreased significantly <italic>（P</italic><0.01）， and the volume of ectopic foci decreased significantly （<italic>P</italic><0.01）. While the level of JAK2，STAT3，p-STAT3，TNF-<italic>α</italic>，VEGF protein in intimal tissue of modified Wenjingtang high and middle dose group decreased significantly （<italic>P</italic><0.05） and the level of TSP-1 protein increased significantly （<italic>P</italic><0.05）. Conclusion:Modified Wenjingtang can inhibit the invasion of ectopic foci in EM rats with kidney deficiency and blood stasis， the mechanism may be related to the intervention of immune barrier and block angiogenesis function mediated by JAK2/STAT3 signaling pathway activation.

A rare case of PLA2R- and THSD7A-positive idiopathic membranous nephropathy / Positividade tecidual dupla para PLA2R e THSD7A em nefropatia membranosa idiopática: raro relato de caso

ABSTRACT Idiopathic membranous nephropathy (IMN) is a frequent cause of nephrotic syndrome in adults. In terms of etiology, the condition may be categorized as primary/idiopathic or secondary. Literature on the pathophysiology of IMN has indicated the presence of autoantibodies (PLA2R and THSD7A) directed against podocyte antigens. The detection of antibodies against a domain favors IMN. The presence of autoantibodies against one of the domains would in theory exclude the possibility of there being autoantibodies against the other domain. However, cases of patients with PLA2R- and THSD7A-positive disease have been recently reported, showing that antibodies against two targets may be concomitantly produced via yet unknown pathophysiological mechanisms. This study reports the case of a 46-year-old male patient with nephrotic-range proteinuria, hematuria, hypoalbuminemia, and hypercholesterolemia submitted to biopsy and histopathology examination (LM, IF, IHC, and EM) eventually diagnosed with PLA2R- and THSD7A-positive IMN associated with IgA nephropathy, stressing our experience with the use of IgG subclasses, PLA2R, and THSD7A in the workup for MN and the relevance of adopting a broad and adequate approach to elucidating and acquiring knowledge of the pathophysiology of IMN.

RESUMO A Nefropatia Membranosa Idiopática (NMi) é uma frequente causa de síndrome nefrótica em adultos e sua etiologia pode ser estratificada em primária/idiopática ou secundária. O conhecimento da fisiopatologia da NMi sugeriu a presença de autoanticorpos (PLA2R e a THSD7A) direcionados contra antígenos existentes nos podócitos. A detecção de anticorpos contra um domínio favorece NMi. A presença de autoanticorpos contra um desses domínios autoexcluiria a possibilidade de autoanticorpos contra o outro domínio; no entanto, recentemente foram descritos casos que apresentaram dupla positividade para PLA2R e THSD7A, comprovando que, por mecanismos fisiopatológicos ainda não conhecidos, raramente pode existir produção concomitante de anticorpos contra os dois alvos. O presente estudo tem por objetivo relatar o caso de um paciente de 46 anos de idade, do sexo masculino, que apresentou quadro de proteinúria nefrótica, hematúria, hipoalbuminemia e hipercolesterolemia submetido a biópsia e exame histopatológico (ML, IF, IHQ e ME), confirmando um caso raro de NMi com positividade dupla para os anticorpos anti-PLA2R e anti-THSD7A e associação à nefropatia por IgA, mostrando nossa experiência com a utilização de subclasses de IgG, PLA2R e THSD7A na rotina laboratorial para a investigação da GNM e enfatizando a importância de uma abordagem ampla para adequada elucidação e conhecimento dos mecanismos fisiopatológicos na NMi.

Evaluation of thrombospondin�gene polymorphisms in corneal allograft rejection in Asian Indian patients

Purpose: To evaluate the frequency and the association of Thrombospondin 1 (THBS1) gene single nucleotide polymorphisms (SNPs) in Asian Indian patients with optical full thickness corneal grafting surgery. Methods: Prospective case朿ontrol analysis of optical penetrating keratoplasty patients with and without immune rejection and controls for genotyping of 3 THBS1 gene SNPs (rs1478604 A>G; rs2228261 C>T; rs2228262 A>G) by Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS PCR). Results: Among 58 patients [45 with immune allograft rejection (DNA isolation was possible in 38 samples) and 13 without immune corneal allograft rejection] and 65 controls, allele frequencies observed for rs1478604 (A>G) are A: 69.7% and 72.6%, G: 30.2% and 27.3%; for rs2228261 (C>T) are T: 70.2% and 62.3%, C: 29.7% and 37.6%; and for rs2228262 (A>G) A: 97.4% and 98.4%; G 2.5% and 1.5% respectively. Genotype frequencies were rs1478604 (A>G) AA: 57.8% and 59.3%, AG 23.6% and 26.5%; GG 18.4% and 14%; for rs2228261 (C>T) TT: 40.5% and 33.8%, TC: 59% and 56.9%, CC: 0% and 9.2%; for rs2228262 (A>G) AA: 94.8% and 96.8%, AG: 5.1% and 3.1% in rejection and controls respectively. The allele and genotype frequency for the 3 described THSB1 SNPs did not show any difference between the corneal graft immune rejection patients and controls. Conclusion: Asian Indian population evaluated for THBS1 gene SNPs by ARMS PCR genotyping in Asian Indian population did not show any genetic association to immune rejection occurrence in our study.

Clinical significance of Mtype phospholipase A2 receptor and thrombospondin Type 1 domaincontaining 7A in primary membranous nephropathy / 中南大学学报(医学版)

OBJECTIVES@#To evaluate the value of thrombospond in Type I domain-containing 7A (THSD7A) and M-type phospholipase A2 receptor (PLA2R) in primary membranous nephropathy (PMN) and to explore the relationship between their antibody levels and prognosis.@*METHODS@#Renal tissues in 128 patients with membranous nephropathy in the Second Xiangya Hospital of Central South University were collected from February 2015 to August 2017, including 108 patients with primary membranous nephropathy (PMN group) and 20 patients with secondary membranous nephropathy (SMN) (SMN group). Indirect immunofluorescence method was used to detect the expression of PLA2R antigen in kidney tissues,and the glomerular expression of THSD7A antigen was examined by immunohistochemistry and indirect immunofluorescence. The serum levels of anti-PLA2R antibodies and THSD7A antibodies were also detected by ELISA. According to the results of PMN examination,the patients were also divided into a PLA2R-related membranous nephropathy group and a THSD7A-related membranous nephropathy group.@*RESULTS@#The positive rate of PLA2R in the renal tissues in the PMN group was higher than that in the SMN group (78% in the PMN group, 35% in the SMN group, <0.01),while the positive rate of anti-PLA2R antibody in the PMN group was also higher than that in the SMN group (50% in the PMN group, 25% in the SMN group, <0.05).The serum level of anti-PLA2R antibody was positively correlated with 24 h urine protein (=0.254, <0.05) and negatively correlated with serum albumin (=-0.236, <0.05). The expression of THSD7A was positive in glomeruli in 7 cases of the PMN group (6%) by immuno-histochemistry, and which was positive in 1case of the SMN group (5%).The serum levels of anti-THSD7A antibody in the PMN group were higher than those in the SMN group [(0.49±0.26) pg/mL in the PMN group,(0.34±0.27) pg/mL in the SMN group, <0.05]. There was no difference in the clinical characteristics between the PLA2R-related membranous nephropathy group and the THSD7A-related membranous nephropathy group.@*CONCLUSIONS@#PLA2R and THSD7A are the target antigen of PMN, and the associated autoantibodies are helpful for the differential diagnosis of PMN. The anti-PLA2R antibody levels can reflect the severity of the disease and evaluate the effect of treatment. The incidence of THSD7A membranous nephropathy is low, and monitoring the serum anti-THSD7A antibody levels can assess patients' condition and predict disease outcome.

Prognostic significance of high expression of thrombospondin 2 in pancreatic cancer and its effect on proliferation and migration of cancer cells / 中国肿瘤生物治疗杂志

@# Objective:To explore the prognostic significance of thrombospondin 2 (THBS2) expression and its effects on the proliferation and migration of pancreatic cancerASPC-1 cells for patients with pancreatic cancer, and to investigate its possible molecular mechanism. Methods: The expression of THBS2 in pancreatic cancer tissues and its effects on overall survival rate in patients were analyzed by online database. THBS2 expression in pancreatic cancerASPC-1 cells was detected by Western Blotting; RNAinterference was used to knockdown the expression of THBS2 inASPC-1 cells, and then the effects of THBS2 knockdown on cell proliferation and migration were detected by MTT and Transwell assays, while its effects on protein expression levels (MMP, E-cadherin,AKT and PI3K) were detected by Wb. Results: Expression of THBS2 in pancreatic cancer tissues was significantly higher than that in normal pancreatic tissues (P<0.01), and the high expression of THBS2 could lead to the decrease of overall survival rate in pancreatic cancer patients. The expression of THBS2 in pancreatic cancer cell lines was significantly up-regulated; however, after interference on the expression of THBS2, the proliferation (P<0.01) and migration ability (P<0.01) of ASPC-1 cells were significantly decreased, and the expression of AKT and PI3K in cells was significantly down-regulated (P<0.01). Conclusion: THBS2 is highly expressed in pancreatic cancer tissues and cells, and is negatively correlated with the prognosis of patients. The mechanism is possibly related with the proliferation and migration of ASPC-1 cells that regulated byAKT/PI3K signaling pathway.

Electroacupuncture down-regulates expression of ADAMTS-4 of intervertebral disc in rats with lumbar intervertebral disc degeneration / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation on the levels of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) protein in the lumbar intervertebral disc tissue and serum in prolapsed lumbar intervertebral disc degeneration (PLIDD) rats, so as to explore its mechanism underlying improvement of intervertebral disc degeneration (IDD). METHODS: A total of 48 Sprague-Dawley rats were divided into sham operation (n＝12), model (n＝18) and EA (n＝18) groups. The PLIDD model was established by puncturing the lumbar discs (L4-L5, L5-L6) with a gauge-22 syringe needle. After modeling, EA stimulation was applied to "Pangguangshu"(BL28), "Zusanli"(ST36) and "Zhijian"for 20 min, 6 times per week for 4 weeks. The lumbar intervertebral disc tissue and blood samples were collected at the 4th, 6th and 8th week after modeling, respectively. The expression of ADAMTS-4 in the lumbar intervertebral disc tissue was detected by immunohistochemistry and Western blot (WB), separately. The content of ADAMTS-4 in the serum was detected by ELISA. RESULTS: Both immunohistochemical stain and WB showed that the expression levels of lumbar ADAMTS-4 at the 4th, 6th and 8th week were significantly up-regulated in the model group relevant to the sham operation group (P<0.01). Following EA treatment, the expression levels of ADAMTS-4 on day 14 and 28 were notably lower in the EA group than in the model group (P<0.05). The serum ADAMTS-4 contents were significantly increased in the model group than in the sham operation group at the 3 time-points (P<0.05, P<0.01), and considerably decreased in the EA group than in the model group on day 28 after EA intervention(P<0.05)．. CONCLUSION: EA can down-regulate the ADAMTS-4 expression of lumbar intervertebral disc tissue in PLIDD rats, which may contribute to its effect in relieving lumbar intervertebral disc degeneration.

Expressions of Versican and ADAMTS-1 in polycysticovary syndrome patients and their clincal significances / 吉林大学学报(医学版)

Objective: To determine the expressions of Versican and a distintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1) in serum in the polycystic ovary syndrome (PCOS) patients, and to clarify their roles in the pathogenesis of PCOS. Methods: A total of 80 patients with PCOS (PCOS group) and 100 healthy women (control group) were selected The heights and body weights of the subjects in two groups were measured; and the body mass index (BMI) was calculated The levels of serum Versican and ADAMTS-1 of the subjects in two groups were measured by enzyme-linked immunosorbent assay (ELISA) method The levels of serum follicle stimulating hormone (FSH), luteinizing hormone (L H), testosterone (T), fasting blood glucose (FBG), fasting insulin of the subjects in two groups were detected; insulin resistance was evaluated according to Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). The correlations between Versican, ADAMTS-1 and the metabolic indexes were analyzed by Pearson linear correlation analysis Results: The serum Versican level of the patients in PCOS group was significantly decreased compared with control group (P = 0 . 004); the serum ADAMTS-1 level was also decreased (P < 0 . 01). The sensitivities, the specificities and the area under receiver operating characteristic (ROC) curve (AUC) of Versican and ADAMTS-1 in prediction of PCOS were 76.74% vs 63. 64% (P = 0 . 018), 52.94% 1)5 40.73% (P = 0 . 009) and 0. 675 (0.550-0.795) vs 0.714 (0 . 6 0 1 - 0. 827) (P = 0 . 032), respectively. The correlation analysis showed that in PCOS group the serum level of Versican of the patients was negatively correlated with FBG (r = - 0 . 7 3 8, P= 0.022), and ADAMTS-1 was negatively correlated with FBG (r = -0.524, P = 0.043). Conclusion: Versican and ADAMTS-1 lowly express in the patients with PCOS. They are negatively correlated with the insulin resistance. They may play inhibitory effects in the occurrence of PCOS.

Diagnostic value of pathological features of atypical membranous nephropathy / 中华肾脏病杂志

Objective To evaluate the diagnostic value of pathological features of atypical membranous nephropathy (AMN). Methods Ninety - one patients with AMN diagnosed by renal biopsy during 2011 and 2017 were enrolled in this study. On the basis of M - type phospholipase A2 receptor (PLA2R) and thrombospondin type - 1 domain - containing 7A protein (THSD7A) by immunohistochemistry, patients were divided into AMN group (25 cases without PLA2R and THSD7A) and idiopathic membranous nephropathy (IMN) group (66 cases with positive PLA2R or THSD7A). The results of immunofluorescence (IF), light microscopy (LM) and electron microscopy (EM) of these two groups were compared, and the parameters with statistical difference were screened out in order to assess their value in the diagnosis of AMN in fourfold table. Results IF results showed that in AMN group the proportions of IgG deposition on capillary wall and mesangial area as well as positive otherIgG subclasses and complement C1q but negative IgG4 were significantly higher than those in IMN group (respectively, 56.0％ vs 12.1％ , 44.0％ vs 0, both P＜0.05). Their diagnostic specificities for AMN were 87.9％ and 100.0％, respectively. However, the positive rates of IgG accompanied with IgA and/or IgM, predominant IgG4 with other IgG subclasses and complement C1q in two groups were not significantly different (all P＞0.05). LM results showed that the proportions of false double track sign on basement membrane and fuchsinophilic proteins under epithelium, endothelium, basement membrane and mesangial region in AMN group were significantly higher than those in IMN group (respectively, 36.0％ vs 0, 44.0％ vs 1.5％, both P＜0.05). Their diagnostic specificities for AMN were 100.0％ and 98.5％ , respectively. However, the scores of mesangial cell proliferation of these two groups showed no significantly difference (P＞0.05). EM results showed that the rate of endothelial electron dense deposits in AMN group was significantly higher than that in IMN group (36.0％ vs 1.5％, P＜0.05), and its diagnostic specificity for AMN was 98.5％. Conclusions IgG deposition on both capillary wall and mesangial area, positive other IgG subclasses and C1q with negative IgG4, false -double contour sign, multi - site fuchsinophilic deposits and endothelial electron dense deposits may help for the AMN diagnosis in the absence of PLA2R and THSD7A related data.