Objective:To investigate the effect of empagliflozin on diabetic kidney disease in db/db mice and the potential mechanisms.Methods:db/db mice were randomly divided into db/db group and Empa group. C57BL/6J mice were used as normal control group. We measured the level of serum biochemistry and inflammatory cytokines. Pathological changes of kidney were observed by pathological staining. The protein expression levels of NLRP3, cleaved caspase-1 and GSDMD were detected.Results:Compared with db/db group, the level of fasting blood glucose, blood lipids, serum biochemistry, and urinary protein-to-creatinine ratio in Empa group were significantly decreased ( P<0.05). HE staining and Masson staining showed that empagliflozin could significantly improve glomerular pyknosis and renal interstitial fibrosis in db/db mice. Meanwhile, the expressions of NLRP3, cleaved caspase-1, and GSDMD protein were down-regulated ( P<0.05). Conclusion:Empagliflozin improves kidney damage in diabetic model mice, and the possible mechanism is to inhibit the cell pyroptosis signal pathway of NLRP3/caspase-1/GSDMD.

Objective To investigate the inhibitory effect on human ACHN cell line and its mice xenograft by using interferon α-1b combined with cyclooxygenase-2 inhibitor and the relevant mechanism in vitro and vivo experiment .Methods ACHN cell and the xenograft mice were devided into 4 groups(IFN-α1b,NS398,IFNα-1b+NS398 and control group).The inhibitory effects were tested by CCK8(Cell Counting Kit 8)assay after AHCN were treated for 24 h and 48 h.The expression of bcl-xl and COX-2 were detected by Western blot .The vol-ume of the xenografts of ACHN cell line and testing the expression of VEGF in xenografts were measured by immunohistochemistry assay .Re-sults Both IFNα-1b and NS398 exerted inhibitory effects on ACHN and this effects showed a rising trend with a increasing concentration of drugs.The combined group was more significant than monotherapy group (P<0.05).Western blot assay showed that IFNα-1b and NS398 downregulated the expression of bcl-xl and COX-2 in ACHN.The combined group was more significant than monotherapy group (P<0.05). The combined group has the greatest inhibitory effects on the xenografts of ACHN cell line compared with monotherapy group and control group(P<0.05).The expression of VEGF in tumor was obiviously inhibited in combined group compared with monotherapy group and con -trol group (P<0.05).Conclusion IFNα-1b combined with NS398 can inhibit the proliferation of ACHN and suppress the tumor growth .

The aim of the research is to investigate the genetic characteristics of Legionella pneumophila serogroup1 (LP1)in Sichuan Province.The sequence-based typing (SBT) and multiple-locus VNTR analysis (MLVA) were used to describe the genetic polymorphism of 42 strains which were isolated from 1989-2016 in Sichuan Province,China.According to the reference,PCR was used to detect the 8-VNTR loci and 7 housekeeping genes respectively.The VNTR results were determined by using capillary electrophoresis,and the SBT results were sequenced and compared with the database of EWGLI.Results showed that totally 42 stains were divided into 8 MLVA types with the advantage types were M08 (47.6 ％) and M07 (23.8 ％).Twelve ST types were obtained with 3 main clonal complex and 2 singleton,including 2 novel ST types,among those,ST1 was the predominant type,accounting for 52.3 ％,following by ST630 (14.2 ％).In conclusion,our results demonstrated MLVA and SBT were both applied to the research for molecular epidemiological investigation of LP1 and showed the high genetic polymorphism and the regional specificity.The results also suggest that the isolates are a potential threat to the public,effective control and prevention strategies are urgently needed.

Objective To explore the clinical application and effect of the transplantation of the cutaneous fibular flap combined with anterolateral thigh flap for the repair of complex tissue defect of the leg. Methods The cutaneous fibular flap combined with anterolateral thigh flap in series connection or parallel connection transfer were applied to repair complex tissue defect of the leg in 36 cases. 10 cases were fresh non-infectious wound 26 cases were delayed infectious wound. The area of wound ranged from 25 cm × 18 cm to 45 cm × 13 cm (36 cm × 16 cm on average). The area of anterolateral thigh flap ranged from 12 cm × 13 cm to 32 cm × 18 cm. The area of the cutaneous fibular flap ranged from 2.0 cm × 1.5 cm to 18.0 cm × 16.0 era. The length of fibular transplantation ranged from 10 cm to 24 cm. 30 cases were combined in parallel connection transfer, 6 cases were combined in series connection transfer, 5 cases were repaired in emergency, 5 cases were repaired in subemergency, 26 cases were repaired in delay. Results All cases were successfully repaired in 36 cases.35 cases were followed up. A mean follow-up was 29 months. Arterial crisis occurred in 1 case, venous crisis occurred in 2 cases 34 flaps survived completely and 2 cutaneous fibular flap survived partially in parallel connection which were later healed by skin transplantation.32 cases were healed in first stage, 4 cases were healed in second stage, (healing time ranged from 12 to 18 days), Bone healing time ranged from 3 to 6 months in fibula transplantation. The Enneking score system was applied to evaluate the leg function. Of the 35 cases, the mean scores was 26 (their scores ranged from 23 to 28).The functions of all supplied regions were not found malfunctional. Conclusion Transplantation of the cutaneous fibular flap combined with anterolateral thigh flap is an optimal method to repair the complex tissue defect of the leg.

Factor VII Deficiency ; diagnosis ; genetics ; Female ; Humans ; Middle Aged ; Pedigree ; Phenotype

Objective To examine the expression of CD93 in serous exosomes from patients with cryptococcal meningitis,fur-ther explore its clinical significance.Methods The 38 experimental serum samples were from patients who received the diag-nosis with cryptococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from November 2012 to De-cember 2016.The diagnosis standardization was that the cerebrospinal fluid dyeing was positive or the culturing was posi-tive.The 38 controls were collected from the health individuals examined at the same time.The exosomes in serum was sepa-rated by the ultracentrifuge method.Magnetic bead-capture combined with the flow cytometry method was used to identify antigens on the surface of exosomes.The protein level of cytokines in serum was quantified by ELISA method.The compari-son of tested values from experimental and controlled groups was measured by two independent samples't test,and the cor-relation between two variates was showed by Pearson coefficient.Results The results of flow cytometry showed that the ex-pression of CD93 on exosomes in the serum samples from experimental group was higher than that in the controlled group, which was(79.11±19.31 vs 23.98±6.56)%,with difference in statistics(t=16.66,P<0.000 1).The expression levels of IFN-γin serum samplefrom experimental and control groups were(39.78±10.77 vs 58.98±16.99)pg/ml,with differ-ence in statistics(t=5.884,P<0.000 1).The expression levels of IL-17 in serum sample from experimental and control groups were(16.32±4.03 vs 3.11±0.87)pg/ml,with difference in statistics(t=19.75,P<0.000 1).The expression lev-els of IL-1β in serum sample from experimental and control groups were(57.12 ± 12.98 vs 13.45 ± 4.78)pg/ml,with difference in statistics(t=19.46,P<0.000 1).The percentage of CD93+exosomes was positively correlative with IFN-γ, IL-17,IL-1β(r=0.488,0.456,0.532,P<0.01)in serum samples from the experimental group and there was statistical difference.Conclusion In the disease process of cryptococcal meningitis,the expression of CD93 on exosomes might involve in the diseases'processes by affecting the CD4+T cells.

Objective To explore the expression level of CD36 in peripheral blood mononuclear cells(PBMCs) from patients with crytococcal meningitis and its clinical significance.Methods The experimental group was the peripheral blood samples from 36 patients diagnosed as crytococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from September 2013 to February 2017,while the control group was the peripheral blood samples from 36 health individuals examined at the same time.PBMCs were separated by density gradient centrifugation method,the relative mRNA expression of CD36,TLR4 in PBMCs was tested by qRT-PCR,the expression level of cytokines such as TNF-α in plasma was tested by ELISA,the comparison of the examined values of experimental and control groups performed by two independent samples' t test,Pearson correlation analysis was performed on the relative mRNA expression of CD36 in PBMCs and cytokines such as TNF-α in experimental group.Results The relative mRNA expression of CD36 in PBMCs of experimental group was 2.01 ± 0.63,the control was 1.49 ± 0.47,and there was statistical difference (t =3.969,P =0.000 2).Pearson analysis results showed that in experimental group CD36 was positively related with the protein expression level of IFN-γ in plasma (r=0.384,P＜0.05) and negatively related with TGF-β(r=-0.487,P＜0.005).Conclusion CD36 might involve in the immunopathology process of crytococcal meningitic by influencing the expression of cytokines such as IFN-γ,TGF-β,and was the potential target for monitoring and treating this disease.

Objective To study the expression of CD69 in exosomes existed in serum from patients with autoimmune pancrea titis (AIP) and primarily discuss its affection on the pathology of disease progression.Methods Serum samples from 35 cases diagnosed as AIP and 35 healthy individuals examined at the same time were collected during October 2012 to December 2016 in Changhai Hospital,they were defined as experimental and control groups,separately.The cytokine levels in serum were measured by ELISA.The expression of surfaced molecules in exosomes existed in serum was tested by magnetic bead conjunct antibody method in flow cytometry.The comparison between the two groups' measurement data was measured by two independent samples' t test,correlation between two variables was showed by Pearson coefficient.Results The flow cy tometry analysis showed that the expression level of CD69 in serum exosomes in experiment and control groups were (85.76 ±19.45 vs 17.01±5.89)％,with statistical difference(t=20.01,P＜0.0001).CD69+ exosomes in experiment group were positively related with serum IL-4,IFN-γ and TGF-β (r 0.456,0.678,0.548,all P＜0.05),while CD69 exosomes were negatively related with serum IL-4,IFN-γ,IL-10 and TGF-β (r=0.589,-0.399,-0.784,-0.657,all P＜0.05).Conclusion The expression of CD69 in exosomes might participate in AIP progression through influencing the function of CD4 +T cell,and it was the potential examined biomarker and therapeutic target.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are confirmed to be expressed in bladder interstitial Cajal-like cells (ICC-LCs), but little is known about their possible role in cystitis-associated bladder dysfunction. The present study aimed to determine the functional role of HCN channels in regulating bladder function under inflammatory conditions. Sixty female wild-type C57BL/6J mice and sixty female HCN1-knockout mice were randomly assigned to experimental and control groups, respectively. Cyclophosphamide (CYP)-induced cystitis models were successfully established in these mice. CYP treatment significantly enhanced HCN channel protein expression and I(h) density and significantly altered bladder HCN1 channel regulatory proteins. Carbachol (CCH) and forskolin (FSK) exerted significant effects on bladder ICC-LC [Ca²⁺]i in CYP-treated wild-type (WT) mice, and HCN1 channel ablation significantly decreased the effects of CCH and FSK on bladder ICC-LC [Ca²⁺]i in both naive and CYP-treated mice. CYP treatment significantly potentiated the spontaneous contractions and CCH (0.001-10 µM)-induced phasic contractions of detrusor strips, and HCN1 channel deletion significantly abated such effects. Finally, we demonstrated that the development of CYP-induced bladder overactivity was reversed in HCN1 -/- mice. Taken together, our results suggest that CYP-induced enhancements of HCN1 channel expression and function in bladder ICC-LCs are essential for cystitis-associated bladder hyperactivity development, indicating that the HCN1 channel may be a novel therapeutic target for managing bladder hyperactivity.
Animals ; Carbachol ; Colforsin ; Cyclophosphamide ; Cystitis ; Female ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels* ; Mice* ; Telocytes* ; Up-Regulation* ; Urinary Bladder*

Objective To analyze the incidence of lung cancer in Qidong , Jiangsu Province of China from 1993 to 2012 . Methods The clinical data of lung cancer from 1993 to 2012 for patients with census registration in Qidong were sorted out from the cancer registration center in Qidong .The annual percent change ( APC) model was used to analyze the trend over time for the incidence of lung cancer .The SAS 9.4 software and the Joinpoint Regression Program 4.3.1.0 were used to implement data analyses . Results A total of 11 895 new cases of lung cancer were diagnosed in Qidong , Jiangsu province from 1993 to 2012 .Among them , 8 629 were male cases and 3 266 were female cases and the median age for these patients was 68 .41 .The crude incidence rate for lung cancer for males was 90 .06/100 000 , which was significantly higher than that for females, i.e., 29.94/100 000(Poisson distribution test, P<0.01). The standardized incidence rate of lung cancer for male was 52 .92/100 000 , which was significantly higher than that for females, i.e.18.52/100 000(Poisson distribution test, P<0.01).The APC for the crude incidence rate for lung cancer was 6 .1 from 1993 to 2012 and the APC for the standardized incidence rate for lung cancer was 5 .5 . Conclusion The incidence of lung cancer increases in Qidong on a yearly basis from 1993 to 2012, with the incidence thereof for males being significantly higher than that for females .