Objective To observe the effects of Angelica and Astragalus ultrafiltration membrane extracts on Keap1/Nrf2 signaling pathway-related factors in diabetes kidney disease(DKD)model rats;To explore its mechanism in treating oxidative stress injury of DKD.Methods Totally 70 SPF-grade male SD rats were randomly divided into 10 rats for the blank group and the remaining 60 rats were fed with high-sugar and high-fat diet,streptozotocin was intraperitoneally injected after 4 weeks to establish the DKD model.The established model rats were randomly divided into the model group,positive control group(GKT137831 30 mg/kg)and TCM high-,medium-and low-dosage groups(Angelica and Astragalus ultrafiltration membrane extracts 6,3,1.5 g/kg),which were orally administered with corresponding drugs.The blank group and model group were given equivalent volume of distilled water orally once daily for 12 consecutive weeks.Random blood glucose,body mass and 24 h urinary total protein content(24 h-UTP)were detected,biochemical assays were conducted to detect serum creatinine(SCr),blood urea nitrogen(BUN),triglycerides(TG),total cholesterol(TC)and albumin(ALB)contents,HE and Masson staining were used to observe the morphology in renal tissue,the contents of reactive oxygen species(ROS),glutathione(GSH)and malondialdehyde(MDA)were measured by ELISA,the protein expressions of nicotinamide adenine dinucleotide phosphate oxidase 4(Nox4),Kelch-like ECH-associated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),glutamate-cysteine ligase modifier subunit(GCLM)and glutathione peroxidase 4(GPX4)in renal tissue were detected by Western blot.Results Compared with the blank group,the random blood glucose at each time point of model group rats significantly increased(P<0.01),the 24 h-UTP significantly increased(P<0.01),the contents of SCr,BUN and serum TG,TC increased(P<0.01),and ALB content decreased(P<0.01);with loose arrangement and disordered structure of renal tissue,consolidation of glomeruli,shedding of renal tubular epithelial cells,and increased deposition of collagen fibers;the contents of serum ROS and MDA were significantly elevated(P<0.01),while GSH contents decreased(P<0.01);the protein expression of Nox4 and Keap1 in renal tissue significantly increased(P<0.01),the expression of Nrf2,GCLM and GPX4 protein significantly decreased(P<0.01).Compared with the model group,the random blood glucose were significantly reduced at 6,8,10,12 weeks in the treatment groups(P<0.01),24 h-UTP decreased(P<0.01),the contents of SCr,BUN,TG,TC in positive control group and TCM high-,medium-dosage group decreased,with increased ALB content(P<0.01,P<0.05);renal tubular and glomerular lesions were improved,and collagen fiber proliferation decreased;serum ROS and MDA contents were reduced,while GSH content increased(P<0.01,P<0.05);the protein expression of Nox4 and Keap1 in renal tissue decreased(P<0.01),and Nrf2,GCLM and GPX4 protein expression increased(P<0.01).Conclusion Angelica and Astragalus ultrafiltration membrane extracts can effectively alleviate injury of renal tissue in DKD rats,and its mechanism may be related to the regulation of the Keap1/Nrf2 signaling pathway and inhibition of oxidative stress response.

Objective To observe the effects of Angelica and Astragalus ultrafiltration membrane extracts on Keap1/Nrf2 signaling pathway-related factors in diabetes kidney disease(DKD)model rats;To explore its mechanism in treating oxidative stress injury of DKD.Methods Totally 70 SPF-grade male SD rats were randomly divided into 10 rats for the blank group and the remaining 60 rats were fed with high-sugar and high-fat diet,streptozotocin was intraperitoneally injected after 4 weeks to establish the DKD model.The established model rats were randomly divided into the model group,positive control group(GKT137831 30 mg/kg)and TCM high-,medium-and low-dosage groups(Angelica and Astragalus ultrafiltration membrane extracts 6,3,1.5 g/kg),which were orally administered with corresponding drugs.The blank group and model group were given equivalent volume of distilled water orally once daily for 12 consecutive weeks.Random blood glucose,body mass and 24 h urinary total protein content(24 h-UTP)were detected,biochemical assays were conducted to detect serum creatinine(SCr),blood urea nitrogen(BUN),triglycerides(TG),total cholesterol(TC)and albumin(ALB)contents,HE and Masson staining were used to observe the morphology in renal tissue,the contents of reactive oxygen species(ROS),glutathione(GSH)and malondialdehyde(MDA)were measured by ELISA,the protein expressions of nicotinamide adenine dinucleotide phosphate oxidase 4(Nox4),Kelch-like ECH-associated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),glutamate-cysteine ligase modifier subunit(GCLM)and glutathione peroxidase 4(GPX4)in renal tissue were detected by Western blot.Results Compared with the blank group,the random blood glucose at each time point of model group rats significantly increased(P<0.01),the 24 h-UTP significantly increased(P<0.01),the contents of SCr,BUN and serum TG,TC increased(P<0.01),and ALB content decreased(P<0.01);with loose arrangement and disordered structure of renal tissue,consolidation of glomeruli,shedding of renal tubular epithelial cells,and increased deposition of collagen fibers;the contents of serum ROS and MDA were significantly elevated(P<0.01),while GSH contents decreased(P<0.01);the protein expression of Nox4 and Keap1 in renal tissue significantly increased(P<0.01),the expression of Nrf2,GCLM and GPX4 protein significantly decreased(P<0.01).Compared with the model group,the random blood glucose were significantly reduced at 6,8,10,12 weeks in the treatment groups(P<0.01),24 h-UTP decreased(P<0.01),the contents of SCr,BUN,TG,TC in positive control group and TCM high-,medium-dosage group decreased,with increased ALB content(P<0.01,P<0.05);renal tubular and glomerular lesions were improved,and collagen fiber proliferation decreased;serum ROS and MDA contents were reduced,while GSH content increased(P<0.01,P<0.05);the protein expression of Nox4 and Keap1 in renal tissue decreased(P<0.01),and Nrf2,GCLM and GPX4 protein expression increased(P<0.01).Conclusion Angelica and Astragalus ultrafiltration membrane extracts can effectively alleviate injury of renal tissue in DKD rats,and its mechanism may be related to the regulation of the Keap1/Nrf2 signaling pathway and inhibition of oxidative stress response.

ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg^-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg^-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.

Objective To explore the effect mechanism of lipophagy in diabetic nephropathy model rats based on intervention of Dangui Hongqi ultrafiltration in diabetic nephropathy model rats.Methods Fifty male Wistar rats of SPF grade were randomly selected 7 rats as the blank group,and the other 43 rats were given a one-time high-dose intraperitoneal injection of streptozotocin combined with high-fat and high-sugar diet to prepare diabetes nephropathy model.The modeling rats were randomly divided into model group,irbesartan group(17.9 mg/kg),and TCM low-,medium-,and high-dosage groups(1.5,3,6 g/kg).The rats were continuously gavaged by corresponding drugs for 12 weeks.Randomized blood glucose,body mass,and 24 h urine protein content were detected;biochemical tests were used for GSP,SCr,BUN,TG,TC,HDL-C,LDL-C,FFA content;HE staining was used to observe morphology changes in renal tissue,Masson staining was used to observe fibrosis changes in renal tissue,renal tissue microstructure changes were observed under transmisson electron microscopy,RT-qPCR was used to detect the mRNA expressions of mTOR and LC3B in renal tissue,and Western blot was used to detect the protein expressions of mTOR,TFEB,p-TFEB,and LC3B in renal tissue.Results Compared with the blank group,the model group rats showed a significant increase in randomized blood glucose(P<0.01),a significant decrease in body mass(P<0.01),a significant increase in 24 h urine protein content(P<0.01),a significant increase in GSP,SCr,BUN,TG,TC,LDL-C,FFA content(P<0.01),and a significant decrease in HDL-C content(P<0.01);renal tubular epithelial cells underwent vacuolar degeneration,cytoplasmic vacuolization,and extensive infiltration of inflammatory cells,collagen fibers increased and were distributed randomly,the number of lipid droplets increased,and autophagosomes decreased;the mTOR mRNA expressions in renal tissue significantly increased(P<0.01),the LC3B mRNA expressions significantly decreased(P<0.01),the mTOR and p-TFEB protein expressions significantly increased(P<0.01),the expression of TFEB,LC3BⅡ protein significantly decreased(P<0.01).Compared with the model group,the randomized blood glucose and 24 h urine protein content of rats in each treatment group were significantly reduced,the body mass significantly increased,and the contents of GSP,SCr,BUN,TG,TC,LDL-C,FFA decreased at 12 week of treatment,while the content of HDL-C increased;the inflammatory cell infiltration,fibrosis,and lipid droplet deposition in renal tissue alleviated to varying degrees;the expressions of mTOR mRNA decreased,the expressions of LC3B mRNA increased,the expressions of mTOR and p-TFEB protein decreased,while the expression of TFEB and LC3BⅡ protein increased.There was a statistically significant difference in the irbesartan group and TCM high-dosage group(P<0.01,P<0.05).Conclusion Dangui Hongqi ultrafiltration can improve the disorder of glucose and lipid metabolism,antagonize heterotopic lipid deposition in the kidney,promote lipid phagocytosis,and delay the process of diabetic nephropathy in model rats.The mechanism may be related to mTOR/TFEB signaling pathway.

Objective To evaluate the effect of Bortezomib in the treatment of secondary plasma cell leukemia in serum M protein and clinical efficacy.Methods20 cases of patients with secondary plasma cell leukemia were randomly divided into the control group and treatment group, 9 cases in the control group,11 cases in the treatment group.The control group was given Prednisolone acetate tablets 60mg/(m2·d),oral, d1~d4 + Melphalan tablets, 8mg/(m2·d),oral, d1~d4+Thalidomide 100~200mg/d, oral, bedtime;on the basis of the control group, the experimental group was treated with Bortezomib for Injection, 1.3mg/(m2·d), intravenous injection, d1、d4、d8、d11.Before and after treatment, compared between the two groups of patients with the ratio of bone marrow plasma cells, serum monoclonicity Immunoglobulin A (IgA), monoclonicity immunoglobulin G (IgG) levels, efficiency of treatment and safety.ResultsAfter treatment, compared with the control group,the plasma cell ratio, the serum levels of IgA and IgG were lower in the experimental group (P<0.05);the treatment efficiency of the experimental group (54.55%) was significantly higher than that of the control group (11.11%), the results were statistically significant.ConclusionThe Bortezomib can significantly reduce the ratio of bone marrow plasma cells and the serum levels of monoclonicity IgA、IgG in patients with secondary plasma cell leukemia, improve clinical efficacy, and the safety was high.

Objective:To study the effects of Radix Angelicae Sinensis ( RASI) on expression of airway MUC5AC and related inflammatory factors in asthmatic mice with Yin deficiency syndrome.Methods:Injecting ovalbumin ( OVA) to sensitize,inhaling OVA to stimulate,using Thyroxin during late stimulation,the asthmatic mouse with Yin deficiency syndrome was established and evaluated through asthmatic behaviors, lung histopathology, active factors ( IL-13, TNF-αand MUC5AC ) in broncho-alveolar lavage fluid (BALF), and MUC5AC expression in lung tissue.Results: 2,4,8 g/kg RASI could reduce asthmatic behaviors score, relieve pathological changes of lung tissue,reduce the contents of IL-13,TNF-αand MUC5AC in BALF,and depress MUC5AC expression in lung tissue ( P<0.05,0.01).In addition,there was a certain synergy between RASI and dexamethasone ( DXM) on depressing the ex-pression of IL-13 and MUC5AC (P<0.05).Conclusion:RASI has certain anti-asthma effect and one of mechanisms is to regulate the MUC5AC expression through inhibit IL-13 and TNF-α.On the expression of IL-13 and MUC5AC,the compatibility of RASI with glu-cocorticoid has some synergy effect.

Objective To establish and evaluate a BALB/c mouse model of anaphylactic asthma with yin-deficiency syndrome. Methods Ovalbumin (OVA) was injected to sensitize and was inhaled to stimulate to replicate the BALB/c mouse model of anaphylactic asthma. Thyroxin was used for gavage during late stimulation to replicate the BALB/c mouse model of asthma with yin-deficiency syndrome. On the basis of monitoring food and water intake and body weight, asthma related indexes, such as asthmatic behaviors, respiratory function, lung histopathology, were detected, and yin-deficiency syndrome related indexes, such as cAMP and cGMP, and pulmonary fluid clearance were examined. Results The BALB/c mouse model of asthma with yin-deficiency syndrome showed obvious asthma symptoms; comprehensive scores of asthmatic behaviors increased significantly;inhaling peak flow, exsufflation peak flow, and tidal volume decreased significantly; lung tissue histopathology showed obvious inflammatory response. Meanwhile, food and water intake of BALB/c mouse model of asthma with yin-deficiency syndrome increased significantly;body weight increased slowly;wet/dry ratio of lung tissue decreased;cAMP and cAMP/cGMP increased, while cGMP decreased, which showed that the mice were in the condition of yin-deficiency and yang-excess. Conclusion Combining thyroxin at the basis of sensitivity induced by OVA is a good pattern to successfully replicate the BALB/c mouse model of anaphylactic asthma with yin-deficiency syndrome.

Objective To observe the effects ofLinggan Wuwei Jiangxin Decoction on contents of cAMP, cAMP dependent PKA and AQP5 of model rats with clod phlegm-fluid lying latent in the lung asthma;To discuss its mechanism.Methods Ovalbumin intraperitoneal injection and atomization motivation + cold stimulation were used to establish models. Ninety rats were randomly divided into blank control group, model group, hexadecadrol group, andLinggan Wuwei Jiangxin Decoction low-, medium-, high-dose groups. All administration groups were treated with relevant medicine. The levels of cAMP, PKA, and AQP5 were detected by ELISA.Results Compared with the blank control group, the levels of cAMP, PKA, and AQP5 of rats in model group decreased obviously (P<0.05). Compared with the model group, the levels of cAMP, PKA, and AQP5 of rats in all administration groups increased (P<0.05), which increased dramatically inLinggan Wuwei Jiangxin Decoction high-dose group.ConclusionLinggan Wuwei Jiangxin Decoction can regulate the normal secretion of cAMP, PKA, and AQP5 in serum of rats with clod phlegm-fluid lying latent in the lung asthma, increase secretion of air passage liquid and decrease mucoprotein concentration, through which plays the role of treating asthma.