Objective:To study the clinical effect of Yang Supplementing Five Returning Decoction(YSFRD) on diabetic peripheral neuropathy. Methods: The curative effect of YSFRD on diabetic peripheral neuropathy and its influence on nerve conduction were comparatively studied. Results: YSFRD was superior to the control group in the curative effect on diabetic peripheral neuropathy ( P

OBJECTlVE To observe the effect of soybean isofIavone( SI)on the expression of nephrin and investigate its protection mechanism in the kidney of diabetic rats. METHODS A diabetic rat modeI was induced after streptozocin(STZ)60 mg·kg-1 was injected into Sprague-DawIey rats. After 72 h,SI 40,120 and 360 mg·kg-1 was ip given to rats in SI groups,respectiveIy,once daiIy,for 8 weeks. The concentration of fasting bIood gIucose( FBG)was determined by a bIood gIucose meter. Urine protein(UP)of 24 h was determined using ELISA. The concentration of serum creatinine(SCr),bIood urea nitrogen(BUN),maIondiaIdehyde(mDA)and superoxide dismutase(SOD)was determined by an automatic biochemicaI anaIyzer. The expression of nephrin,tumor necrosis factor-α(TNF-α)and NF-κB protein in renaI tissue was measured by Western bIotting,respectiveIy. RESULTS Compared with normaI controI group,the concentration of FBG,24 h UP,BUN,SCr and mDA in modeI group was aII siginificantIy increased(P﹤0.05,P﹤0.01),but SOD was decreased( P﹤0.01).Compared with modeI group,BUN in SI 40,120 and 360 mg·kg-1 groups decreased from(24.3±6.3)mmoI·L-1 to(16.8±4.9), (13.4±5.4)and(7.7±1.2)mmoI·L-1 ,respectiveIy,24 h UP in SI 360 mg·kg-1 group decreased from (1.26±0.45)mg to(0.88±0.15)mg( P﹤0.01),SCr decreased from(56.4±8.4)μmoI·L-1 to(35.3± 4.3)μmoI·L-1(P﹤0.01),mDA decreased from(8.32±1.40)μmoI·L-1 to(5.33±0.95)μmoI·L-1(P﹤0.01), but SOD in SI 120 and 360 mg·kg-1 groups increased from(125.5 ±2.4)kU·L-1 to 144.2 ±9.2 and (169.2±3.2)kU·L-1(P﹤0.01). Compared with SI 120 mg·kg-1 ,SCr,BUN and mDA in SI 360 mg·kg-1 group were aII siginificantIy Iower(P﹤0.05,P﹤0.01),but SOD was higher(P﹤0.01). Compared with normaI controI group,the expression of nephrin protein greatIy decreased(P﹤0.01),whiIe the expres-sion of TNF-α protein greatIy increased in modeI group( P ﹤0.01). Compared with modeI group,the expression of nephrin protein greatIy increased in SI 40,120 and 360 mg·kg-1 groups(P﹤0.01),whiIe the expression of TNF-α and NF-κB protein greatIy decreased in SI 120 and 360 mg·kg-1 groups(P﹤0.01). CONCLUSlON The mechanism by which SI protects the kidney may be partIy reIated to anti-infIammation,antioxidation and increasing the expression of nephrin in diabetic rats.

Objective To analyze the characteristics of breast medullary carcinoma in CEUS and to compare with pathologic features.Methods Morphologic characteristics of 13 breast medullary carcinomas in CEUS were analyzed.The diameter of mass before and after CEUS were compared.Parameters from time-intensity curves of masses were analyzed in contrast with peripheral breast parenchyma.All the results from CEUS analysis were compared with pathological manifestations.Results Breast medullary carcinoma was characterized as irregular shape (n=10),clear margins (n=11) and uniform enhancement (n=11) in CEUS.These characteristics were in accordance with their morphologic characters in pathology.The diameter of mass before and after CEUS had no significant defference (P=0.61),which was in accordance with expansive growth in pathology.In contrast with peripheral breast parenchyma,the arrival time and time to peak of breast medullary carcinoma were significantly shorter (P=0.034,0.021),and peak enhancement intensity was significantly stronger (P=0.005),which were in accordance with the increased vascular density and their uniform distribution,big arteries at the margin of masses in pathology.Conclusion Breast medullary carcinoma has distinguished characteristics in CEUS,which are in accordance with characters in pathology,and can be used as the basis in clinical diagnosis and differential diagnosis of breast medullary carcinoma.

Objective To investigate the mechanism of Smad ubiquitination-related factor 2 (Smurf2)neddylation. Methods The Smurf2 protein level was tested by overexpression of Nedd8,while the method of immunoprecipitation(IP) and Western blotting were used to analyz Smurf2-Nedd8 modification.The GST-pulldown experiment was conducted to prove protein interactions.The protein was obtained by grinding mouse tissue and Western blotting was used to test the protein expression level.Results Over expression of Nedd8 could lead to the down regulation of the Smurf2′s protein level.Smurf1 and Smurf2 could interact in the GST-pulldown experiment. Smurf1 could enhance Smurf2-Nedd8 modification.The Smurf2′s protein level was up-regulated in mouse tissue of Smurf1 C426A.Conclusion There is some relationship and also difference between Smurf1 and Smurf2.Smurf1 can enhance Smurf2-Nedd8 modification.

Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.

Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.

Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells.Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad 3 were designed and synthesized .pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques .Overexpression of Smad 3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A 549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed .Overexpression of Smad 3 significantly up-regulated the migration rate of A549 and HeLa cells.Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate .Conclusions Smad3 promotes cell migration of A549 and HeLa cells.

Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration.Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively.After 7 days,the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening,enrichment and expansion.The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot.Transwell assay was used to assess the effect of Smurf1 silencing on cell migration.Results The stable cell lines with Smurf1 silencing are constructed successfully.Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay.Conclusion Smurf1 promotes cell migration.

Summary: The levels of serum TNF-α and IL-8 in the patients with allergic asthma during acute attack period and remission period, and the effects of glucocorticoid (GC) on them were investigated. By using ELISA, the levels of TNF-α and IL-8 were detected in the healthy volunteers (group C, n=40), the patients with allergic asthma (n=40) during acute attack period (group A) and remission period (group B) and those taking GC for a week (n=28). The results were compared among them. It was found that the levels of TNF-α and IL-8 in group A were higher than in group B and group C. In the patients subject to GC therapy, the levels of TNF-α and IL-8 were decreased as compared with those in group A. In group B, the level of TNF-α was higher than in group C, but there was no significant difference in the level of IL-8 between group B and group C. It was concluded that the inflammatory cytokines, TNF-α and IL-8, played important roles in the bronchus allergic inflammation. GC could reduce the levels of serum TNF-α and IL-8 to exert the anti-inflammatory effects.

Objective To establish a patient-derived xenografts (PDX) mouse model of liver cancer (LC) and to explore its role in precision medicine.Methods PDX model was established by subcutaneous implantation of tumor tissues in NCG mice.The morphological structure of tumor tissue was exaimed using HE staining.Fifteen BALB/c nude mice were subcutaneously inoculated with tumor cell suspension from the PDX models.The xenograft mice were randomly divided into 5-fluorouracil (5-FU) group, sorafenib group and negative control group.The tumor volume and body weight of the tumor-bearing mice were measured regularly, the tumor inhibition rate was calculated and the curative effect was evaluated.Results The success rate was 33.3% (6/18) in the establishment of liver cancer PDX mouse model, and the model well retained the characteristics of the primary tumor.In one case of PDX mouse model, the tumor inhibition rates of 5-FU and sorafenib group were 63.7% and 29.6%, with a statistically significant differece between them (P< 0.05), and there was no significant difference between the sorafenib group and negative control group, consistent with clinical observation.Conclusions The PDX mouse model of liver cancer can maintain the histological structure of primary tumor, and can be applied to precision medicine for patients with liver cancer.