Objective To find out a more rational pathological classification criteria for renal injury in patients with type 2 diabetes mellitus. Methods The renal clinicopathological features of forty-nine type 2 diabetic patients with maeroalbuminuria were collected and were compared retrospectively. The patients without diabetic renal disease were excluded. According to the pathological features, the patients were divided into two groups: typical diabetic glomerulopathy (DG) and atypical diabetes-related renal disease (ADRD). Results The renal biopsy revealed DG accounted for 59.2% of the patients, while the remaining 40.8% presented atypical renal injury defined as ADRD. In DG group, volume fraction of mesangium per glomerulus, glomerular basement membrane width, atrophic tubules index, intersititium injury index and prevalence of hyalinization of renal arteriole were higher; podocyte density per glomerulus was lower; duration of type 2 diabetes was longer; the level of fast blood glucose, systolic blood pressure, mean arterial pressure, proteinuria and prevalence of diabetic retinopathy (DR) were higher; glomerular filtration rate (GFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher; dyslipidemia was more severe. GFR was negatively correlated with glomerular global sclerosis rate in both DG and ADRD group. Proteinuria was positively correlated with volume fraction of mesangium per glomerulus in DG. No correlation between proteinuria and pathological features was found in ADRD. DR (94.8%) and duration of type 2 diabetes over five years (90.7%) had high negative predictive value for DG. Conclusions Renal injuries in type 2 diabetes patients are heterogeneous. ADRD is an atypical renal injury in type 2 diabetes patients whieh is different from DG. DR and duration of diabetes are more helpful in predicting DG separating from ADRD.

Objective To study the pathologic pattern and clinical feature of type 2 diabetes mellitus complicated with chronic kidney diseases (CKD). Methods Clinicopathological features of 155 type 2 diabetic patients complicated with CKD were collected and analyzed retrospectively. The patients were divided into four groups: typical diabetic glomerulopathy (DG),atypical diabetes-related renal disease (ADRD), non-diabetic renal diseases (NDRD) and DG complicated with NDRD. Results Renal biopsies revealed DG accounted for 18.7% of the patients, ADRD accounted for 12.9%, NDRD accounted for 60.0%, and DG complicated with NDRD accounted for 8.4%. In DG group, duration of type 2 diabetes was longer;the level of fast blood glucose, systolic blood pressure, mean arterial pressure and prevalence of diabetic retinopathy (DR) were higher;proteinuria was heavier and evaluated glomerular filtration rate (eGFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher;dyslipidemia was more severe. Gross hematuria and acute renal insufficiency could be only found in NDRD group.Without DR, duration of diabetes under 5 years, gross hematuria, acute renal insufficiency,evidences of autoimmune diseases and proteinuria≥3.5 g/24 h but eGFR ≥60 ml/min were specific valuable predictors for NDRD. Conclusions Renal injuries in type 2 diabetic patients are structural heterogeneous, in which NDRD is more common and is different from ADRD and DG.Renal biopsy should be considered when type 2 diabetic patients complicated with CKD present at least one characteristic as follows: duration of diabetes under 5 years, without DR, history of gross hematuria, acute decrease of renal function, evidences of autoimmune diseases and proteinuria ≥ 3.5 g/24 h but eGFR ≥ 60 ml/min.

Objective To establish a culture protocol for Th17 cells in vitro and evaluate the effect of curcumin on the differentiation and functions of Th17 cells and explore the related mechanisms. Methods Splenic CD4+CD25- T cells of C57BL/6 mice were isolated and purified with magnetic bead methods, and were co-cultured with plate-bound anti-CD3 and anti-CD28 antibodies and with transforming growth factor (TGF)-β, interleukin (IL)-6, IL-23, anti-interferon-γ antibody and anti-IL-2 antibody for Th17-polarization. The cultured Th17 cells were allocated into four groups: the control group, in which T cells were cultured on the basis of the above protocol; the low-concentration curcumin (CM-L, 5 μmol/L) and high-concentration (CM-H, 25 μmol/L) curcumin groups, and sirolimus (SRL, 100 ng/ml) group. The proportion of Th17 cells was detected with flow cytometry, and the mRNA expression of retinoic acid-related orphan receptor (ROR)γt, IL-17A, IL-21 was examined with real-time quantitative polymerase chain reaction. Finally, the protein expression of RORγt and the phosphorylation levels of signal transducer and activator of transcription (STAT) 3 were measured using western blotting analysis. Results The proportion of Th17 cells in the freshly isolated CD4+CD25- T cells was (3.1 ±0.4)%, whereas the proportion of the cells cultured with the protocol could get [(54.1±3.4)%, P<0.01]. As compared with the control group, the Th17 cells proportion in CM-L, CM-H and SRL groups was significantly decreased [CM-L (40.3±2.8)%, CM-H (25.8±2.3)%, SRL (25.0±2.0)%versus the control (54.1±3.4)%, t=6.15, 12.63, 12.97, P<0.01], and no marked difference was seen between CM-H group and SRL group (ft>0.05). Moreover, the mRNA levels of RORγt, IL-17A and IL-21, the protein expression of RORγt and the phosphorylation levels of STAT 3 in CM-L, CM-H and SRL groups were also lower than those in the control group (IL-17A mRNA: CM-L 0.81±0.05, CM-H 0.61±0.05, SRL 0.58±0.05, Control 1.01 ±0.11, t=4.81, 8.52, 8.89; IL-21 mRNA: CM-L 0.73±0.06, CM-H 0.49±0.03,SRL 0.59±0.03, Control 1.12±0.11, t=5.98, 9.22, 7.95, P<0.01). The mRNA level of IL-21 in the CM-H group was lower than that in the SRL group (P<0.05). Conclusion In vitro, curcumin can inhibit the differentiaton of splenic CD4+CD25- T cells into Th17 cells in the setting of Th 17-polarization and inhibit the expression of IL-17A and IL-21 mRNA, which is associated with the inhibitive effect of curcumin on RORγt expression and STAT 3 phosphorylation.

Objective To examine the expression of Cath D,SP-A and GLUT-1 in patients with bronchiogenic carcinoma.Methods A total of 41 patients were included in the study,10 of whom received the histological diagnosis of small cell lung cancer(SCLC).The other 28 were squamous cell carcinoma(SC) and 3 were inflammation.And all the samples were taken from the patients′ tunica mucosa bronchiorum through bronchofibroscope,then we detected the cathepsin D,SP-A and GLUT-1 following SP immunohistochemistry.Results ①Cath D: 6 samples(60%) in group SCLC were negtive expression(-6),4(40%)were moderately positive(++4),4(14%) were negative(-4),2(7%) were positive(+2),8(28%) were moderately positive(++8) and 14(50%) were intensive positive(+++14) in the other group.SCLC was significant different from SC in expressing cathepsin D(P

Objective To investigate the effect of curcumin on autophagy and inflammatory response in mice subject to renal ischemia reperfusion.Method Forty male C57BL/6 mice were randomly divided into four groups (n =10 each):sham-operated (SO) group (Abdominal incision was made to expose the kidneys,bilateral renal pedicle dissociated and the abdomen sewed),ischemia reperfusion (IR) group,curcumin (CM) group (given CM 10 mg/kg) and 3-methyladenine (3-MA) group (given 3-MA 15 mg/kg).Six and 24 h after reperfusion,renal function was tested by determining the serum creatinine (Scr) and blood urea nitrogen (BUN) levels,and the morphological changes in the kidney tissue were observed.The expression of LC3,Beclin 1,Rab7 and LAMP2 in kidney tissue was detected by fluorescence quantitative RT-PCR and Western blotting.Fluorescence quantitative reverse transcription PCR and enzyme-linked immunosorbent assay were used to examine the expression of IL-6,IL-10,IL-17 and TNF-α in kidney tissue and serum.Result As compared with IR group and 3-MA group,the Scr and BUN levels in the CM group were significantly decreased (P＜0.01),and the renal morphological changes were improved significantly (P＜0.01).The mRNA and protein expression of LC3,Beclin 1,Rab7 and LAMP2 was significantly increased in kidney tissue,and the expression of IL-6,IL-17 and TNF-α was reduced,while IL-10 was increased (P＜ 0.01) in the CM group as compared with IR group and 3-MA group.Conclusion Curcumin possesses a protective effect against renal ischemia reperfusion injury in mice,which is probably mediated by promoting autophagy and subsequently inhibiting inflammatory response.

Objective:To compare the safety and efficacy of ticagrelor and clopidogrel in dual antiplatelet therapy for stent-assisted embolization of unruptured intracranial aneurysms.Methods:Patients with unruptured intracranial aneurysms received stent-assisted embolization in the Department of Neurosurgery, Linyi People's Hospital from January 2021 to June 2022 were retrospectively included. According to the preprocedural dual antiplatelet therapy scheme, they were divided into aspirin+clopidogrel group (clopidogrel group) and aspirin+ticagrelor group (ticagrelor group). The incidence of ischemic and bleeding events was compared between the clopidogrel group and the ticagrelor group at 3 months after procedure. Multivariate logistic regression model was used to analyze independent risk factors for postprocedural ischemic and bleeding events. Results:A total of 195 patients were included. Their age was 58.15±10.11 years and 75 were males (38.5%). There was no statistically significant difference in the incidence of bleeding events (12.8% vs. 5.9%) and ischemic events (14.9% vs. 18.8%) at 3 months after procedure between the ticagrelor group ( n=94) and the clopidogrel group ( n=101). Multivariate logistic regression analysis showed that smoking (odds ratio [ OR] 6.085; 95% confidence interval [ CI] 1.589-13.012; P=0.019], hypertension ( OR 4.547, 95% CI 1.589-13.012; P=0.005), aneurysm at the branch vessel ( OR 3.089, 95% CI 1.122-8.504; P=0.029), and the use of flow diverter ( OR 3.111, 95% CI 1.062-9.110; P=0.038) were the independent risk factors for postprocedural ischemic events. Triglycerides might be an independent risk factor for postprocedural bleeding events ( OR 1.435, 95% CI 0.989-2.082; P=0.057), but did not reach statistical significance. Conclusions:In dual antiplatelet therapy for stent-assisted embolization of unruptured intracranial aneurysms, ticagrelor and clopidogrel have the same safety and efficacy.

OBJECTIVETo study mRNA expression of receptor activator nuclear factor kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG) in peri-implant tissue during unloading period.

METHODSAn animal model of dental implant was established in 6 male Beagle dogs of 1-2 years old. Bone remodeling was tested at 3, 7, 15, 30, 60 and 90 days since the placement of implants. RANKL and OPG mRNA expression were quantified by real-time polymerase chain reaction (PCR). Then mandibular bones were taken out and the morphological changes were observed by X-ray, bone tissue was tested by immunohistochemistry stain.

RESULTSThe most prominent period of bone remodeling occurred at 7th day after the placement of implants. The expression of RANKL and OPG increased in a time-dependent manner in both soft and hard tissue. After 7 days they gradually decreased.

CONCLUSIONRANKL and OPG can express in soft tissue, and the changing tendency is consistent with the change of bone remodeling, it indicates that RANKL and OPG play an important role in the bone remodeling.

Animals ; Bone Remodeling ; Bone and Bones ; Carrier Proteins ; Dogs ; Male ; NF-kappa B ; Osteoprotegerin ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B

Objective:To investigate the efficacy and safety of Tubridge flow diverter (TFD) in the treatment of unruptured intracranial aneurysms (UIAs).Methods:The clinical data of patients with UIAs treated with TFD in the Department of Neurosurgery, Linyi People's Hospital from July 2021 to July 2023 were retrospectively analyzed. The modified Rankin Scale (mRS) was used to evaluate functional outcomes during postoperative follow-up. Digital subtraction angiography (DSA) follow-up was performed at least 3 months after procedure, and the occlusion degree of the aneurysm was assessed with the O'Kelly-Marotta (OKM) grading system.Results:A total of 52 patients (71 aneurysms) were included and 54 TFDs were successfully implanted. Forty-one patients (78.8%) were treated with TFD alone, while 11 (21.2%) were treated with TFD combined with coil embolization. One patient experienced in-stent thrombosis during the procedure, and the symptoms disappeared after medical treatment. Postoperative hemorrhage and ischemic complications occurred in each 2 cases, respectively. No patient died during the follow-up period. DSA follow-up was performed on a total of 45 aneurysms in 34 patients. The followed-up time was 8.3±3.1 months (range, 3-15 months), including 2 OKM grade A (4.4%), 11 grade B (24.4%), 1 grade C (2.2%), and 31 grade D (68.9%). At the last follow-up, 51 patients had good outcomes (mRS score 0-2), and 1 patient had poor outcome (mRS score 4).Conclusion:TFD for UIAs has a high complete occlusion rate and a low complication rate.

Objective:To explore the predictive values of tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) in donor sera and lavage fluid on delayed graft function (DGF) in donation after circulatory death (DCD) kidney transplant recipients.Methods:A total of 33 eligible kidney donors and 33 corresponding recipients were recruited. Preoperative serum and renal perfusion fluid samples of donors were collected to determine the levels of TIMP-2 and IGFBP7. Patients were grouped according to whether DGF occurred after kidney transplantation and measured indicators analyzed. Independent sample t test was utilized for comparing the groups with normal distribution measurement data. And χ2 test was employed for comparing the groups with normal distribution counting data and Mann-Whitney test for comparing the groups with non-normal distribution measurement data. Receiver operating characteristic (ROC) curve and area under curve (AUC) were used for evaluating the diagnostic efficacy of indicators. Results:In donor-DGF group, lavage fluid TIMP-2, product of lavage fluid TIMP-2 and IGFBP7 (TIMP-2×IGFBP7), serum IGFBP7 and product of serum TIMP-2 and IGFBP7 (TIMP-2×IGFBP7) were higher than those in donor-non-DGF group ( P<0.05). The AUC of TIMP-2, TIMP-2×IGFBP7, serum IGFBP7 and serum TIMP-2×IGFBP7 in the diagnosis of DGF were 0.753 (95%CI 0.546~0.959), 0.747 (95%CI 0.510~0.984), 0.824 (95%CI 0.615~1.000) and 0.852 (95%CI 0.660~1.000) respectively. Conclusions:Donor serum IGFBP7, donor serum TIMP-2×IGFBP7, lavage fluid TIMP-2 and lavage fluid TIMP-2×IGFBP7 may be used for predicting the occurrence of early DGF after kidney transplantation. Among them, serum TIMP-2×IGFBP7 has the highest diagnostic efficiency and may be an excellent predictor of DGF occurrence.

Objective:To design a modified S1PR3 specific agonist, GPS-725.017, and investigate its protective effect on acute lung injury by promoting macrophage clearance of bacteria.Methods:A short peptide derived from the intracellular region of S1PR3 receptor was named GPS725.017, which was modified with norleucine (Nle) and myristicacid (myr) at its N terminus. Mice were divided into the sham operation group, solvent group and GPS-725.017 treatment group. The acute lung injury model was induced by endotracheal injection of E. coli (5×10 6 CFU), and the experimental group was treated with GPS-725.017 (10 mg/kg). The 48-h survival rate of mice was recorded. After 5 h of modeling, the bacterial load and inflammatory cytokines in peripheral blood and lung were detected, and Vps34 protein content in alveolar macrophages was determined by Western blot. After 12-h of modeling, lung tissues were collected for H&E staining and pathological scores. Results:Compared with the solvent group, the survival rate of mice in the GPS-725.017 treatment group was significantly improved ( P<0.01), the bacterial CFU in blood and alveolar lavage fluid was significantly lower than that in the solvent group ( P<0.001), and the levels of TNF-α and IL-1β in blood and alveolar lavage fluid were significantly lower than those in the solvent group ( P<0.001). Western blot showed that the expression level of Vps34 protein in alveolar macrophages was significantly higher than that in the solvent group ( P<0.01). Histopathology result showed that the pathological damage of lung in the treatment group was significantly less than that in the solvent group ( P<0.001). Conclusions:The modified synthetic S1PR3 specific agonist GPS-725.017 could specifically activate the S1PR3 receptor on the membrane of alveolar macrophages and up-regulate the expression level of intracellular Vps34 protein, which can promote the removal of bacteria in alveolar macrophages, significantly reduce the degree of lung injury and improve the survival rate in ALI mice.