Objective To observe the effects of Hedysarum Polybotrys polysaccharide(HPS)on the glucose metabolism of small intestinal smooth muscle in rats with diabetic gastroparesis(DGP)of spleen qi deficiency syndrome;To explore its mechanism based on AMPK/GLUT4 signaling pathway.Methods Totally 72 male Wistar rats were randomly divided into 12 in the blank group and the remaining 60 rats were assigned to the model group.The DGP spleen qi deficiency syndrome model was replicated by intraperitoneal injection of streptozotocin+irregular feeding with high-fat and high sugar feed combined with swimming exhaustion method.The model rats were randomly divided into model group,metformin group and HPS high-,medium-and low-dosage groups.The metformin group was given 100 mg/kg metformin hydrochloride by gavage,while the HPS high-,medium-and low-dosage groups were given 200,100 and 50 mg/kg HPS by gavage,respectively.The blank group and model group were given purified water by gavage once a day for 8 consecutive weeks.The gastric emptying rate and small intestine propulsion rate in rats were detected,HE staining was used to observe the smooth muscle morphology of gastric antrum and ileal tissue,immunofluorescence staining was used to detect the expression of glucagon like peptide-1(GLP-1)in ileal tissue,ELISA was used to detect the contents of adiponectin(APN),glucagon(Glu)and insulin(INS)in serum,Western blot was used to detect the protein expressions of glucose transporter(GLUT)4,GLUT1,AMP-activated protein kinase(AMPK)and p-AMPK in ileal tissue.Results Compared with the blank group,the model group rats showed significantly increased random blood glucose,significantly decreased body mass,gastric emptying rate and small intestinal propulsion rate(P<0.01);the edema in the submucosal layer,loose arrangement of connective tissue,infiltration of a small number of lymphocytes,granulocytes and macrophages,reduced number of goblet cells in the ileal tissue,shedding of intestinal villous epithelial cells and widening of the gap between the submucosal layer and the lamina propria;the expression of GLP-1 in ileal tissue was significantly decreased(P<0.05),the contents of serum APN and INS were significantly decreased,and the content of Glu significantly increased(P<0.01),the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue were significantly decreased,while the expression of GLUT1 protein significantly increased(P<0.01).Compared with the model group,rats in metformin group and HPS high-dosage group showed a random decrease in blood glucose,an increase in body mass,gastric emptying rate,and small intestine propulsion rate(P<0.05,P<0.01);a more regular arrangement of gastric antral tissue cells,a reduction of shedding cells and edema,the smooth muscle structure of the ileal tissue was relatively intact,with evenly distributed cells and reduced infiltration of inflammatory cells;the expression of GLP-1 in ileal tissue increased,the contents of serum APN and INS increased,and the content of Glu decreased,the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue increased,while the expression of GLUT1 protein decreased(P<0.05,P<0.01).Conclusion HPS may up-regulate GLUT4 and down-regulate GLUT1 expression through activates AMPK,promotes glucose uptake and utilization by small intestinal smooth muscle,and improves glucose metabolism of DGP rats with spleen qi deficiency syndrome.

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 Hedysarum Polybotrys polysaccharide(HPS)on the glucose metabolism of small intestinal smooth muscle in rats with diabetic gastroparesis(DGP)of spleen qi deficiency syndrome;To explore its mechanism based on AMPK/GLUT4 signaling pathway.Methods Totally 72 male Wistar rats were randomly divided into 12 in the blank group and the remaining 60 rats were assigned to the model group.The DGP spleen qi deficiency syndrome model was replicated by intraperitoneal injection of streptozotocin+irregular feeding with high-fat and high sugar feed combined with swimming exhaustion method.The model rats were randomly divided into model group,metformin group and HPS high-,medium-and low-dosage groups.The metformin group was given 100 mg/kg metformin hydrochloride by gavage,while the HPS high-,medium-and low-dosage groups were given 200,100 and 50 mg/kg HPS by gavage,respectively.The blank group and model group were given purified water by gavage once a day for 8 consecutive weeks.The gastric emptying rate and small intestine propulsion rate in rats were detected,HE staining was used to observe the smooth muscle morphology of gastric antrum and ileal tissue,immunofluorescence staining was used to detect the expression of glucagon like peptide-1(GLP-1)in ileal tissue,ELISA was used to detect the contents of adiponectin(APN),glucagon(Glu)and insulin(INS)in serum,Western blot was used to detect the protein expressions of glucose transporter(GLUT)4,GLUT1,AMP-activated protein kinase(AMPK)and p-AMPK in ileal tissue.Results Compared with the blank group,the model group rats showed significantly increased random blood glucose,significantly decreased body mass,gastric emptying rate and small intestinal propulsion rate(P<0.01);the edema in the submucosal layer,loose arrangement of connective tissue,infiltration of a small number of lymphocytes,granulocytes and macrophages,reduced number of goblet cells in the ileal tissue,shedding of intestinal villous epithelial cells and widening of the gap between the submucosal layer and the lamina propria;the expression of GLP-1 in ileal tissue was significantly decreased(P<0.05),the contents of serum APN and INS were significantly decreased,and the content of Glu significantly increased(P<0.01),the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue were significantly decreased,while the expression of GLUT1 protein significantly increased(P<0.01).Compared with the model group,rats in metformin group and HPS high-dosage group showed a random decrease in blood glucose,an increase in body mass,gastric emptying rate,and small intestine propulsion rate(P<0.05,P<0.01);a more regular arrangement of gastric antral tissue cells,a reduction of shedding cells and edema,the smooth muscle structure of the ileal tissue was relatively intact,with evenly distributed cells and reduced infiltration of inflammatory cells;the expression of GLP-1 in ileal tissue increased,the contents of serum APN and INS increased,and the content of Glu decreased,the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue increased,while the expression of GLUT1 protein decreased(P<0.05,P<0.01).Conclusion HPS may up-regulate GLUT4 and down-regulate GLUT1 expression through activates AMPK,promotes glucose uptake and utilization by small intestinal smooth muscle,and improves glucose metabolism of DGP rats with spleen qi deficiency syndrome.

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.

Diabetic gastroparesis （DGP） is a common diabetic neuropathy that affects the normal function of gastric motility and emptying. Clinically， it often manifests as abdominal distension， nausea and vomiting， early satiety， dyspepsia， etc. The pathogenesis of DGP is multifactorial， closely related to many factors， such as chronic hyperglycemia， neuropathy， autonomic nervous system disorders， inflammation， and oxidative stress. These factors can interact with each other， leading to delayed gastric emptying and the occurrence of related symptoms. Traditional Chinese medicine （TCM） has significant advantages in the prevention and treatment of DGP， including a long history， remarkable efficacy， individualized treatment， diverse therapeutic formulations， and improvement in the quality of life. Additionally， TCM is known for its low adverse reactions， good tolerance， and multi-targeted effects， making it an important approach in the management of DGP. Previous research has found that the main mechanisms of Chinese medicine in the prevention and treatment of DGP include the regulation of gastrointestinal hormones， inhibition of inflammatory responses， reduction of oxidative stress， enhancement of interstitial cells of Cajal activity， inhibition of pyroptosis， and modulation of related signaling pathways such as stem cell factor （SCF）/cellular growth factor receptor （c-Kit）， adenosine monophosphate-activated protein kinase （AMPK）， Ras homologous genome member A （RhoA）/Rho-associated coiled-coil forming kinase （ROCK）. This article primarily summarized the research progress on Chinese medicine in preventing and treating DGP through the inhibition of inflammatory responses， reduction of oxidative stress， enhancement of interstitial cells of Cajal activity， inhibition of pyroptosis， and regulation of related signaling pathways， aiming to provide a reference and basis for further research on the application value of Chinese medicine in the prevention and treatment of DGP.

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 study the epidemic and etiologic characteristics of influenza virus(Flu)and respiratory syncytial virus (RSV) infections in outpatient influenza-like illness(ILI)cases of children under 10 years of age in Gansu province in 2018 And to provide evidences for diagnosis, treatment, control and prevention of Flu and RSV infections in Gansu province.Methods:A total of 8 559 pharyngeal swab samples of ILI outpatients cases were tested with real-time fluorescent PCR to detect Flu, 3 436 of which were detected by RT-PCR for RSV.Results:Of the 8559 specimens, 934 (10.91%) samples were positive for Flu. Among them, 431 were positive for H1N1, 70 were positive for H3N2, 428 were positive for Flu B, 5 were mixed; 320 of the 3436 (9.31%, ) samples were positive for RSV. There were significant differences in the positive rates of Flu and RSV among 14 cities and prefectures ( χ2=56.99, χ2=263.34, Р< 0.01). Influenza reached its peak in January 2018 with a positive rate of 50.70%. Flu B/Yamagata strain (53.53%) and A H1N1 (39.93%) were prevalent simultaneously. The peak of RSV epidemic was from February to April, the positive rate was 13.98%. The RSV positive rate of children under 5 years of age was 10.11%, higher than that of children over 5 years of age was 6.94%. There was statistical significance ( χ2=7.67, Р<0.01). Conclusions:RSV and influenza viruses are the main pathogens in ILI cases of children under 10 years of age. There are epidemic peaks in winter and spring every year. It is suggested that the monitoring of RSV and the development and application of vaccine should be strengthened.