BACKGROUND:Diabetic osteoporosis is gaining public attention.However,few studies have reported the effect of a high-glucose environment on the osteogenic differentiation of human umbilical cord mesenchymal stem cells and the corresponding therapeutic strategies. OBJECTIVE:To investigate whether vitamin D3 can restore the osteogenic differentiation potential of human umbilical cord mesenchymal stem cells in a high-glucose environment. METHODS:The viability of human umbilical cord mesenchymal stem cells was detected by CCK-8 assay to screen the appropriate vitamin D3 intervention concentration.Under the high-glucose environment,RT-qPCR,western blot assay,immunofluorescence,JC-1 mitochondrial membrane potential,alizarin red staining,and β-galactosidase staining were used to evaluate the osteogenic differentiation potential,intracellular reactive oxygen species accumulation,mitochondrial membrane potential alteration,and cell senescence of human umbilical cord mesenchymal stem cells after vitamin D3 intervention.The underlying mechanism was also discussed. RESULTS AND CONCLUSION:(1)Vitamin D3 significantly promoted the proliferation of human umbilical cord mesenchymal stem cells in the range of 0.1 μmol/L to 1 mmol/L.(2)High-glucose environment down-regulated the mRNA and protein level expressions of osteogenic-related genes α1-I collagen,alkaline phosphatase,Runt-associated transcription factor 2,and osteocalcin in human umbilical cord mesenchymal stem cells,which induced oxidative stress and cellular senescence.(3)Vitamin D3 at an intervention concentration of 10 μmol/L significantly restored the osteogenic phenotype of human umbilical cord mesenchymal stem cells under high-glucose conditions and attenuated intracellular oxidative stress and cellular senescence by activating the Nrf2/HO-1 signaling pathway.(4)These findings suggested that the osteogenic differentiation ability of human umbilical cord mesenchymal stem cells was reduced in the high-glucose environment,and vitamin D3 could partially improve their osteogenic differentiation ability and reduce cell damage.

BACKGROUND:Osteoporosis has a high incidence,leading to fracture and other complications.However,existing drugs have great side effects and are difficult to meet the clinical application. OBJECTIVE:To explore the effect and potential mechanism of fucoxanthin on osteoporosis induced by glucocorticoid. METHODS:Primary rat osteoblasts were inoculated in 6-well plates.When the cell fusion reached 80%,the cells were divided into four groups:the control group was cultured alone for 24 hours,the glucocorticoid group was intervened with dexamethasone for 24 hours,the fucoxanthin group was intervened with fucoxanthin for 24 hours,and the glucocorticoid + fucoxanthin group was intervened with dexamethasone and fucoxanthin at the same time for 24 hours.After intervention,cell proliferation,apoptosis,intracellular reactive oxygen species level,and protein expression of apoptosis-related proteins,bone formation-related proteins,and nuclear factor erythroid-2-related factor 2 were detected. RESULTS AND CONCLUSION:Cell counting kit-8 results showed that the cell viability was decreased in the glucocorticoid group compared with the control group(P＜0.05)but increased in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).JC-1 mitochondrial membrane potential staining and flow cytometry assay showed that the percentage of apoptosis increased in the glucocorticoid group compared with the control group(P＜0.05)but decreased in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).Western blot assay showed that compared with the control group,the protein expression of BAX and cleaved poly(ADP-ribose)polymerase was elevated in the glucocorticoid group(P＜0.05),and the protein expression of BCL2,type Ⅰ collagen α1 peptide chain,alkaline phosphatase,osteocalcin,and RUNX2 was decreased in the glucocorticoid group(P＜0.05).Compared with the glucocorticoid group,the protein expression of BAX and cleaved poly(ADP-ribose)polymerase was decreased(P＜0.05),and the protein expression of BCL2,type Ⅰ collagen α1 peptide chain,alkaline phosphatase,osteocalcin,and RUNX2 was elevated(P＜0.05)in the glucocorticoid+fucoxanthin group.Fluorescent probe assay showed an increase in reactive oxygen species level in the glucocorticoid group compared with the control group(P＜0.05)and a decrease in reactive oxygen species level in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).Immunofluorescence staining and western blot assay showed that the protein expression of nuclear factor erythroid-2-related factor 2 in the glucocorticoid group was decreased compared with that in the control group(P＜0.05);and the protein expression of nuclear factor erythroid-2-related factor 2 in the glucocorticoid+fucoxanthin group was elevated compared with that in the glucocorticoid group(P＜0.05).To conclude,fucoxanthin can improve glucocorticoid-induced osteoblast apoptosis and the expression of bone formation-related molecules by activating nuclear factor erythroid-2-related factor 2.

Objective:To investigate the effects and the mechanism of renal protection of curcumin on oxidative stress and lipid metabolism in diabetic rats.Methods:60 Wistar rats were randomly divided into normal control group ( n=20) and diabetic model preparation group ( n=40). Rats in the normal control group were fed with normal diet, while rats in the diabetes model group were fed with high sugar and high fat diet. After 4 weeks of feeding, rats in the diabetic model preparation group were injected with streptozotocin (60 mg/kg) intraperitoneally once. A total of 35 rats were successfully modeled and then randomly divided into diabetic model group ( n=17) and curcumin group ( n=18). The diabetic model group continued to be fed with high sugar and high fat diet, and the curcumin group was given high sugar and high fat diet and 200 mg/(kg·d) curcumin (1% carboxymethyl fiber sodium prepared into suspension) intervention every day; while the normal control group and the diabetic model group were given 1% carboxymethyl fiber sodium of the same volume. The body weight, kidney index, blood glucose , serum creatinine, urea nitrogen, blood lipid and 24-hour urinary microalbumin excretion rate (24 h UAER) of rats in each group were measured. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and m-alondialdehyde (MDA) in serum and kidney tissues were measured. Results:Compared with the control group, the levels of the body weight, kidney index, blood glucose, serum creatinine, urea nitrogen, triglyceride, total cholesterol and 24 h UAER in the diabetic model group increased significantly. The level of MDA in serum and kidney tissues increased significantly ( P<0.05), while the levels of SOD, CAT and GSH-Px activities in serum and kidney tissues decreased significantly ( P<0.05). Compared with diabetic model group, the levels of the body weight increased, while the kidney index, blood glucose, serum creatinine, urea nitrogen , triglyceride, total cholesterol and 24 h UAER in the curcumin group decreased significantly, the level of MDA in serum and kidney tissues decreased significantly ( P<0.05); the levels of SOD, CAT and GSH-Px in serum and kidney tissues increased significantly ( P<0.05) in the curcumin group. Conclusions:Curcumin may play a renal protective role in diabetic rats by inhibiting oxidative stress in serum and kidney and reducing blood lipid.

OBJECTIVETo investigate the feasibility and safety of membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection for advanced distal gastric cancer.

METHODSThe clinical data of 41 patients with advanced distal gastric cancer who underwent laparoscopic gastrectomy using membrane-based right-sided approach for laparoscopic suprapancreatic lymph node dissection at the Department of Gastrointestinal Surgery, Zhongshan Hospital of Xiamen University from January 2016 to January 2018 were retrospectively analyzed. There were 24 males and 17 females with a mean age of 56.8 years and a mean body mass index of 22.6 kg/m². Membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection contained 4 steps briefly: (1) dissection of mesenteria above the head of pancreas: the tri-junction of pancreas-duodenum was cut to expose and identify the logo of Benz;clearance of the membrane of No.5a was performed towards the left, and then expanded to the posterior layer of No.12a. (2) dissection of the V shape dorsal mesogastrium: membrane bridge at splenic artery trunk root was cut; in suprapancreatic space, clearance was performed towards to the left to the middle of the splenic artery trunk and expanded to the posterior Tolds plane upwards to the posterior phrenic angle and retroperitoneal esophagus, then the surrounding tissue of anterior abdominal aorta. (3) dissection of the U shape mesenteria:membrane bridge at common hepatic artery root was cut; mesentery was separated; the left gastric vein was freed and ligated at its root; in posterior pancreatic space, the mesentery of No.7, No.9 and No.8 was dissected in turns; the left gastric artery was high ligated and cut; the portal vein and posterior dorsal mesogastrium Toldt plane was routinely exposed; clearance was performed to right for No.8a and upward to the hepatic portal meeting at posterior mesentery No.12 plane. (4) dissection of the upper triangular area of pylorus: the trigone mesentery was cut along the upper edge of the pylorus; No.12a was swept upward along the gastric ventral mesentery; the upper boundary(No.8a) on the right side of the U-shaped membrane was joined. Intraoperative and postoperative presentations were analyzed.

RESULTSLaparoscopic gastrectomy for advanced distal gastric cancer with membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection was successfully carried out in all the 41 patients. Distal gastric mesenteria en bloc resection was successfully performed. The operation time was (145.2±25.4) minutes and intraoperative blood loss was (53.3±18.3) ml without massive bleeding and severe complication. Number of lymph nodes dissected was 41.1±6.4, and number of suprapancreatic lymph node dissected was 23.3±3.7 without residual cancer at cut margin by pathology. Postoperative drainage volume was (65.8±21.7) ml; time to withdraw of catheter was (7.0±1.7) days; time to fluid intake was (3.5±1.8) days; postoperative hospital stay was (10.4±2.8) days; time to postoperative anal exhaust was (3.3±1.1) days. No complications, such as chyle leakage, postoperative massive bleeding, anastomotic leakage, abdominal cavity infection or gastroplegia occurred within 30 days after surgery.

CONCLUSIONMembrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection for advanced distal gastric cancer can achieve en bloc resection and conform to the radical principle of oncology, and is safe and feasible.

Objective To investigate the value of KIM-1 and NGAL in predicting the early diabetes nephropathy(DN).Meth-ods 132 cases of type 1 and type 2 diabetic patients were recruited in this study,patients were divided into four groups based on the urine albumin/creatinine and serum creatinine levels,as follows:normal albuminuria group (45 cases),mi-croalbuminuria group (36 cases),clinical albuminuria group (30 cases),renal failure group (21 cases),65 healthy subjects were recruited as control group.Urinary KIM-1 and NGAL levels were measured by enzyme linked immunosorbent assay (ELISA).Results With the progress of DN,urinary KIM-1 and NGAL levels gradually increased (P<0.05),the urinary KIM-1/Ucr level was positively correlated with ACR (r=0.822,P<0.01),and the urinary NGAL/Ucr was positively cor-related with ACR (r=0.842,P<0.01).Conclusion The urinary KIM-1 and NGAL level can predict the occurrence of ear-ly DN,and also monitor the progression of DN.

Objective To investigate the effects of whole grain-bean mixed staple food on intestinal microecology,anthropometric,and metabolic parameters of obese people.Methods Totally 87 obese people were divided with random number table into whole grain-bean mixed staple food group (test group) and refined grains staple food group (control group).Body weight (BW),body mass index (BMI),waist circumference (WC),hip circumference (HC),waist/hip ratio (WHR),fasting plasma glucose (FPG),triglyceride (TG),total cholesterol (TC),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),and colony count of intestinal flora (bacillus bifidus,lactobacillus,clostridium perfringens,enterobacteria,enterococcus,bacteroides) were measured at baseline and 3 months after intervention.Results Ten people were excluded from this research,6 in the test group and 4 in the control group.Mter 3 months of dietary intervention,BW,BMI,WC,HC,and WHR were significantly lower in the test group than in the control group [(69.45 ± 7.07) kgvs.(72.42 ±5.79) kg,P=0.000; (26.08 ±1.48) kg/m2 vs.(27.27 ±0.81) kg/m2,P=0.000; (82.48±9.30) cm vs.(86.96±4.93) cm,P =0.000; (90.08 ±6.57) cm vs.(92.42 ±6.67) cm,P =0.000; 0.92 ±0.11vs.0.95 ±0.10,P =0.003].The levels of FPG,TC,and LDL-C in the test group were significantly lower than those in the control group [(4.92 ± 0.75) mmoL/L vs.(5.41 ± 1.21) mmoL/L,P=0.037; (3.85±1.13) mmoL/L vs.(4.38 ±1.26) mmoL/L,P=0.046; (3.55 ±1.04) mmoL/L vs.(4.16 ± 1.40) rnmoL/L,P =0.033] ; the level of HDL-C in the test group was significantly higher than that in the control group [(1.13 ± 0.37) mmoL/L vs.(0.92 ± 0.26) mmoL/L,P =0.004].The colony counts of intestinal bacillus bifidus,lactobacillus,and bacteroides in the test group were significantly higher than in the control group [(7.94 ± 0.58) lg CFU/g vs.(6.95 ± 0.38) lg CFU/g,P =0.000 ; (7.67 ± 0.46) lg CFU/g vs.(6.96 ± 0.57) lg CFU/g,P =0.000 ; (5.53 ± 0.44) lg CFU/g vs.(4.80 ±0.23) lg CFU/g,P =0.000],while the colony count of clostridium perfringens was significantly lower than in the control group [(4.40 ± 0.49) lg CFU/g vs.(5.11 ± 0.63) lg CFU/g,P =0.000].Conclusions In obese people,whole grain-bean mixed staple food can improve anthropometric parameters,some lipid metabolic parameters,and intestinal flora.The underlying mechanism may involve promoting the survival and proliferation of probiotics,thereby improving glucose and lipid metabolism.

Objective To compare the differences of atria and atrial myocyte structure in patients with atrial fibrillation (AF) and sinus rhythm(SR) .To evaluate the influence of AF on cardiac function .Methods 79 patients without heart failure undergoing car-diopulmonary bypass surgery were divided into the AF group (n=39) and the SR group(n=40) .Echocardiography was performed for analysis of left ventricular end diastolic dimension(LVDd) ,left ventricular end-systolic dimension(LVDs) ,left ventricular poste-rior wall(LVPW) ,interventricular septum(IVS) and left atrial diameter(LAD) .Part of left atrial appendages was taken freshly for HE staining in order to observe atrial tissue structure .Results LVDd ,LVDs ,LVPW and IVS of the AF group were lower than that of the SR group .But LAD of the AF group was higher than the SR group .There was statistical significance in IVS and LAD between the two groups(P<0 .05) ,and there were no significant differences in other aspects (P>0 .05) .Compared with the SR group ,the AF group had thinner myocardial atrophy ,more obvious fibrosis ,smaller nucleus and darker HE staining .Conclusion The incidence of AF was mainly in rheumatic heart disease .The development of AF was mostly accompanied with the enlargement of LAD and the change of atrial tissue structure which showed that AF can reduce cardiac function .

Objective To investigate the relationship of urinary liver-type fatty-acid binding protein and progression of ne-phropathy in diabetic patients .Methods A total of 132 cases of type 1 and type 2 diabetic patients were recruited in this study , and were divided into four groups based on the urine albumin /creatinine and serum creatinine levels , including normal albuminuria group ( n =45), microalbuminuria group ( n =36), clinical albuminuria group ( n =30), and renal failure group ( n =21), Additional 65 healthy subjects were recruited as control group .The levels of urinary liver-type fatty-acid binding protein ( L-FABP) were measured with enzyme-linked immunosorbent assay (ELISA), and serum creatinine, urine creatinine, Liver function, hemoglobin, glycated he-moglobin (HbA1c), albumin (Alb), and 24h urinary protein were measured by respective biochemical or immunological methods . Results With the progression of diabetic nephropathy , the level of urinary L-FABP was gradually increased ( P <0.01 ) , subgroup analysis showed that the level of urine L-FABP of normal albuminuria group was significantly higher than that of healthy group ( P <0.05 ) .The level of urine L-FABP of microalbuminuria group was significantly higher than that of normal albuminuria group ( P <0.01 ) .The level of urine L-FABP of clinical albuminuria group was significantly higher than that of microalbuminuria group ( P <0.01 ) .The level of urine L-FABP of renal failure group was significantly higher than that of clinical albuminuria group ( P <0.01 ) . The levels of urinary L-FABP was negatively correlated with estimated glomerular filtration rate ( eGFR) ( r =-0.812 , P <0.01 ) , was positively correlated with Scr ( r =0.901, P <0.01), and was no correlation with HbA1c ( P >0.05).Conclusions The uri-nary L-FABP level can predict the occurrence of early diabetic nephropathy , and also monitor the progression of diabetic nephropathy .

OBJECTIVE: To explore the expression profile of Ephrin-B2 in the ischemic penumbra after transient focal cerebral ischemia in rats, and to clarify the mechanism of Ephrin-B2 triggering angiogenesis. METHODS: Sprague-Dawley rats were randomly divided into a normal group, a sham operation group and ischemic-reperfusion 1, 3, 7, 14, and 28 d groups. Suture-occluded method was used to establish the focal middle cerebral artery occlusion model and the ischemic brain was reperfused 2 h after the occlusion. Western blot and quantitative real-time reverse-transcription polymerase chain reaction were used to detect the dynamic expression profile of Ephrin-B2 in the penumbra cortex. Double immunofluorescence was used to speculate the location and the co-expression of Ephrin-B2 in blood vessels, neurons and astrocytes. Microvessel density was quantified by the number of CD31+ cells. Rats were subjected to neurologic functional tests by modified neurological severity scores (mNSS) before sacrifice. RESULTS: Compared with the sham group, Ephrin-B2 protein and mRNA level of the penumbra cortex in the ischemic group increased 3 days (P<0.05) after the reperfusion, peaked at day 7 and 14 (P<0.01), and declined at day 28. Double immunofluorescence indicated that Ehprin-b2 was expressed in the neurons, blood vessels and astrocytes; mNSS peaked at day 7, and gradually declined at day 14. The microvessel density of penumbra cortex in the ischemic group increased 3 days (P<0.05) after the reperfusion, peaked at day 14 (P<0.01), and gradually declined at 48 h. CONCLUSION Cerebral ischemia reperfusion induces the over-expression of Ephrin-B2, with a dynamic trend, suggesting that Ehprin-b2 may improve post-stroke functional recovery by enhancing angiogenesis and neurogenesis.
Animals ; Astrocytes ; metabolism ; Brain ; pathology ; Brain Ischemia ; metabolism ; Cerebral Cortex ; metabolism ; Ephrin-B2 ; metabolism ; Infarction, Middle Cerebral Artery ; Ischemic Attack, Transient ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

Objective To evaluate the bone healing effect of vascularized tissue-engineered bone induced by endothelial progenitor cells(EPCs)in repair of large segmental radius defects in rabbits.Methods A total of 68 healthy New Zealand white rabbits were enrolled in the study and randomized into three groups,ie,experimental group(EPCs group):EPCs plus bone marrow mesenchymal stem cells (BMSCs)plus decalcified bone matrix(DBM);control group:BMSCs plus DBM;sham control group:pure DBM.Materials mentioned above were implanted into middle radius defects for 15 mm.At 12 and 16 weeks post-operatively,X-ray test,bone mineral density test,histological light microscopic test,osteocalcin immunohistochemical staining test and biomechanical test were carried out.Results Growth and plasticity of callus,speed of medullary cavity recanalization,bone healing speed and biomechanical intensity in the experimental group were all significantly better than those of control group.Conclusions Vascularized tissue-engineered bone induced by EPCs has strong osteegenic ability,can accelerate bone healing and hence is an effective method for repair of large segmental bone defects.