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Objective@#To explore the allogeneic mouse adipose-derived mesenchymal stem cell (ADSC)-microporous sheep acellular dermal matrix (ADM) on healing of wound with full-thickness skin defect in mouse and the related mechanism.@*Methods@#One Kunming mouse was sacrificed by cervical dislocation to collect adipose tissue from inguinal region. Mouse ADSCs were isolated from the adipose tissue and cultured in vitro. Cells of the third passage were identified by cell adipogenic and osteogenic differentiation. The expressions of CD73, CD90, CD105, and CD34 were analyzed by flow cytometry. After one sheep was sacrificed, microporous sheep ADM was prepared from sheep back using decellularization method and freezing-thawing method. A 12 mm diameter, round, full-thickness skin defect wound was made on the back of each one of 36 Kunming mice. The wounds were covered by microporous sheep ADM. The mice were divided into group ADSC and control (C) group with 18 mice in each group according to the random number table after surgery. A volume of 0.2 mL DMEM/F12 culture medium containing 1×106 ADSCs was injected between microporous sheep ADM and wound of mice in group ADSC. While 0.2 mL DMEM/F12 culture medium was injected between microporous sheep ADM and wound of mice in group C. On post surgery day (PSD) 12 and 17, wound healing rates of mice in the 2 groups were calculated. On PSD 7, 12, and 17, wound vascularization of mice in the 2 groups was observed under reverse irradiation of backlight. On PSD 7, 12, and 17, the wound granulation tissue of mice in group ADSC was observed by hematoxylin and eosin staining. On PSD 7, the thicknesses of granulation tissue of mice in the 2 groups was measured. On PSD 12 and 17, expressions of VEGF in wounds of mice in the 2 groups were detected by immunohistochemical method. The sample number was 6 in each group at each time point in the above experiments. Data were processed with t test and analysis of variance of factorial design.@*Results@#(1) After 7 days of adipogenic induction, lipid droplet was observed in cytoplasm using oil red O staining. After 21 days of osteogenic induction, black deposits of calcium salts were detected using silver nitrate staining. Expression rates of CD73, CD90, CD105, and CD34 in cells were 97.82%, 99.32%, 97.35%, and 5.88% respectively. The cells were identified as ADSCs. (2) The wound healing rates of mice in group ADSC on PSD 12 and 17 [(78±6)%, (98±3)%] were significantly higher than those in group C [(60±9)%, (90±4)%, t=4.26, 4.46, P<0.01]. (3) On PSD 7, no vessel obviously grew into the center of wounds of mice in the 2 groups, while the granulation tissue has covered the wounds of mice in group ADSC. On PSD 12, the vessels were more abundant in wounds of mice in group ADSC than those in group C. On PSD 17, big vessels crossing the whole wounds was observed in wounds of mice in group ADSC, while big vessels were observed without crossing the whole wounds in wounds of mice in group C. (4) The wounds were covered with thin granulation tissue on PSD 7, and the granulation tissue began to thicken on PSD 12 and were covered by epidermis on PSD 17 in wounds of mice in group ADSC. On PSD 7, the granulation tissue in wounds of mice in group ADSC [(0.62±0.05) mm] was significantly thicker than that in group C [ (0.31±0.04) mm, t=12.27, P<0.01]. (5) On PSD 12 and 17, expressions of VEGF in wounds of mice in group ADSC [(80.7±2.2), (0.98±0.03)/mm2] were significantly than those in group C [(59.5±2.4), (81.5±2.6)/mm2, t=15.95, 14.14, P<0.01].@*Conclusions@#Allogeneic mouse ADSC-microporous sheep ADM can accelerate angiogenesis and growth of granulation tissue, thus promoting wound healing, which may be due to the increase of expression of VEGF.
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Objective To monitor and compare the breathing mechanics on PC,VC and PRVC during pneumoperitoneum,and to discuss the significance of the clinic use of PRVC.Method Ninety laparoscopic cholecystectomy patients were equally divided into 3 groups (PC,VC,PRVC).Levels of PES,PAWM,PAP,PaCO2,ETCO2,TV MAP and HR were detected before pneumoperitoneum,and at 5,10,15 and 20 minutes postpneumoperitoneum.Results Pneumoperitoneum made three respiratory patterns with different levels of PAWM,PAP,and PES.PES post-pneumoperitoneum in the VC model was obviously higher than that in the PC and PRVC group.At 10 min post-pneumoperitoneum,levels of PaCO2 and ETCO2 increased obviously in the PC and VC group(P ＜ 0.05).Levels of PaCO2 and ETCO2 were increased in the PC group,but TV level post-pneumoperitoneum was significantly lower than that in the other two groups (P ＜ 0.05).Level of PaCO2 and ETCO2 were increased in the PC and VC group post-pneumoperitoneum,along with increases of MAP and HR (P ＜ 0.05).Levels of MAP and HR in the PRVC group post-pneumoperitoneum were significantly lower than those in the PC and VC group (P ＜ 0.05).Conclusion PRVC mode can effectively reduce the increases of pneumoperitoneum-induced PAWM,PAP,PES,without the unusual increase of PaCO2 and ETCO2 during surgeries,contributing to the stability of vital signs of perioperative patients.
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Objective To evaluate the effect of resveratrol on the growth of an established A431 xenogratt tumor in nude mice.Methods The model of human skin squamous cell carcinoma was established by inoculating A431 cells in log-phase growth into the left axillary fossa of Balb/c (nu/nu) nude mice.After 7-8 days,60 mice bearing human A431 skin squamous cell carcinoma xenografts were randomly and equally divided into 6 groups:blank control group receiving no treatment,negative control group treated with intraperitoneal sodium chloride physiological solution,positive control group treated with intraperitoneal cyclophosphamide,high-,medium-and low-dose resveratrol groups treated with intraperitoneal resveratrol of 40,20 and 10 μg per gram body weight per day,respectively.Tumor size was measured at a 4-day interval during the treatment course.After 14-day treatment,the mice were sacrificed.Xenograft tumors were removed from these mice and subjected to weight measurement,pathological examination by hematoxylin and eosin (HE) staining and apoptosis detection by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Western blot was conducted to quantify the protein expression of apoptosis-related factors,including phosphorylated extracellular signal-regulated protein kinase (p-ERK),p53 and caspase 3.Data were processed by SPSS 13.0 software,and statistical analysis was carried out by analysis of variance and Pearson correlation analysis.Results By the end of treatment,the xenograft tumor volume was (1153.56 ± 255.41) mm3,(1001.69 ± 115.08) mm3,(1206.80 ± 175.88) mm3,(1342.28 ± 211.12) mm3,(1642.34 ± 225.85) mm3 and (1564.32 ± 156.49) mm3,and the weight was (1.84 ±0.30) g,(1.72 ± 0.39) g,(1.96 ± 0.40) g,(2.67 ± 0.73) g,(3.16 ± 0.52) g,and (3.33 ± 0.59) g,respectively in the positive control group,high-,medium-and low-dose resveratrol group,negative control group and blank control group.Significant differences were observed in the xenograft tumor volume (F =16.00,P ＜ 0.05) and weight (F =19.15,P ＜ 0.05) among the 6 groups.According to the tumor weight,the growth of tumor was inhibited by 45.57％,37.97％ and 15.51％ respectively in the high-,medium-and low-dose resveratrol groups.Increased apoptotic index was observed in the positive control group,high-,medium-and low-dose resveratrol groups compared with the negative control group and blank control group (36.79 ± 8.86,33.15 ± 6.00,18.09 ±3.92 and 10.53 ± 4.20 vs.3.87 ± 1.63 and 2.73 ± 1.61,F =93.26,P ＜ 0.05).Analysis of variance showed that the protein expressions of p-ERK,p53 and caspase 3 were all higher in the three resveratrol groups than in the negative control group and blank control group (F =6.65,6.78,11.56,respectively,all P ＜ 0.05).The protein expression of p53 was statistically correlated with p-ERK (r =0.68,P ＜ 0.05) and caspase 3 (r =0.56,P ＜0.05).Conclusions Resveratrol shows an inhibitory effect on the growth of human A431 skin squamous cell carcinoma xenografts in nude mice,likely by increasing p53 expression and inducing tumor cell apoptosis via the activation of MAPK/ERK pathway.
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Objective To explore the changes on BDNF mRNA in transected spinal cord and associated motor cortex and skeleton muscle following cord injury in rats.Methods 20 adult Sprague Dawleys rats were performed spinal cord transected operation at T11 level,then rats in each group(n=5) were sacrificed on 1,3,7 and 14 days post operation respectively.Other 5 rats were used as normal control without operation.The tissues from the rostral,caudal segments near injury site,cerebral cortex and linea obliqua tibiae were harvested.Total RNA was extracted with Trizol reagent separately.The BDNF mRNA expression in each group was detected by RT-PCR.Results(1)BDNF positive bands were seen in the tissues of the rostral,caudal segments near injury site,cerebral cortex and linea obliqua tibiae.Moreover,BDNF level in cerebral cortex is more than in the spinal cord at normal control(P