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.

Objective To study possible clinical relationships of myeloperoxidase (MPO) and carotid atherosclerosis in diabetic patients on maintenance hemodialysis(MHD).Methods Levels of plasma MPO and serum elastase (ELT) of diabetic patients on MHD were determined by enzyme-linked immunosorbent assay.Carotid intima media thickness(CIMT),quantity and area of carotid plaque were measured by B-mode ultrasonography.Results Plasma MPO was significantly higher after hemodialysis(HD) than before HD(414.6 (198.9,671.5) μg/L vs 176.4 (69.9,243.3) μg/L,Z =-4.51,P ＜ 0.01).There was no significant difference of serum ELT before and after HD(198.0(146.9,270.5) μg/L vs 230.9(40.2,308.0) μg/L,Z =-1.87,P ＞ 0.05).There was positive correlation between the age and CIMT,quantity or area of carotid plaqueor(correlation coefficient:0.764,0.416 and 0.446 respectively,P ＜ 0.01 or P ＜ 0.05),There was positive correlation between MPO level before HD and age,Levels of serum ELT before HD,or CIMT(correlation coefficient:0.592,0.476 and 0.810 respectively,P ＜ 0.01).There was positive correlation between MPO level after HD and levels of serum ELT after HD (correlation coefficient:0.364,P ＜ 0.05).There was no correlation between the levels of plasma MPO after HD and CIMT,quantity or area of carotid plaqueor (P ＞ 0.05).Conclusion (1) Age and MPO level before HD contribute to carotid atherosclerosis in patients on MHD.(2)There was no relation between higher MPO level after HD and carotid atherosclerosis.

Objective To investigate the G protein-coupled receptor kinase 2 (GRK 2) level in peripheral blood lymphocytes with cardiac func-tion in elderly patients with acute myocardial infarction. Methods This study enrolled 40 patients with acute ST-segment elevation myo-cardial infarction (STEMI) and 40 patients with unstable angina. All patients were 65 years or older. Cardiac function was evaluated by echocardiography, and the GRK 2 level in peripheral blood lymphocytes was measured. Patients with STEMI were followed up for 2 years. Results The GRK 2 level in peripheral blood lymphocytes was significantly higher in patients with STEMI than in patients with unstable angina, and was negatively correlated with left ventricular ejection fraction, cardiac output, stroke volume, and left ventricular fractional shortening. The GRK 2 level was significantly elevated in some patients with acute STEMI and poor cardiac function. Conclusions In-creased GRK 2 level in patients with acute STEMI may contribute to poor myocardial systolic function and myocardial remodeling. Meas-urement of the GRK 2 level in peripheral blood lymphocytes may assist in the evaluation of cardiac function and myocardial remodeling in elderly patients with acute STEMI.

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

Objective To investigate the association between donor complement factor component 7 (C7) rs6876739 gene polymorphisms and risk of early bacterial infection following orthotopic liver transplantation (OLT).Methods A total of 113 patients who had undergone OLT in Shanghai General Hospital between July 2007 and January 2011 were included.A single nucleotide polymorphism (SNP),donor C7 rs6876739 was genotyped and analyzed together with clinical data.Results We demortstrated that donor C7 rs6876739 CC genotype had higher risk of early bacterial infection than TT genotype following OLT (55.6％ vs.26.5％,P =0.021).The multivariate logistic regression analysis revealed that gender (P =0.018),biliary complications (P =0.018),ICU stay after LT (P＜0.001) and donor C7 rs6876739 genotype (P =0.001) were identified as independent factors of early bacterial infection.Conclusion Donor C7 rs6876739 genotype polymorphism is associated with early bacterial infection following OLT and may be a new marker of risk for the development of potentially serious bacterial infection after liver transplantation.

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.

Acute promyelocytic leukemia (APL) is characterized by the generation of the prototypic promyelocytic leukemia-retinoicacid receptor alpha (PML-RARα), an oncogenic fusion protein due to chromosomal translocation. In a human myeloid cell line,PML-RARα is cleaved by neutrophil elastase (NE) to produce the mutational PML [nuclear localization signal (NLS) deletion andRARα (NLS-RARα, containing NLS of PML), both of which may play an important role in APL pathogenesis. The yeast two-hybridtechnique was used to screen the intracellular proteins interacting with NLS-RARα, which may be involved in NLS-RARα signaling. The NLS-RARα coding sequence was amplified by polymerase chain reaction method and was cloned into the bait plasmid pGBKT7vector, which, after the confirmation by sequencing, was transformed into yeast AH109 and the subsequent expression of bait plasmidwas proved by Western-blot. The transformed yeast AH109 was mated with yeast Y187 (containing leukemia cDNA library plasmidspACT2) in medium. Diploid yeast was plated on synthetic dropout nutrient medium containing X-α-gal for screening. After beingreintroduced into yeast AH109 and sequenced to verify the expression of ORF, eight positive colonies were obtained, among whichonecontaining JTV-1 was cloned. The interaction between NLS-RARα and JTV-1 was further supported by indirect immunofluorescence,GST pull-down and co-immunoprecipitation, respectively. These findings brought some new clues for the further exploration ofNLS-RARα signaling to APL.

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)/mm^2, 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.

Objective:To evaluate the effect of adductor canal block(ACB)and local infiltration anesthesia(LIA)around the knee joint on inflammatory responses in the patients undergoing total knee arthroplasty(TKA).Methods:Sixty American Society of Anesthesiologists physical status Ⅱor Ⅲ patients of both sexes, aged 54-76 yr, scheduled for elective TKA, were divided into 2 groups ( n=30 each) using a random number table method: ACB group (group A) and ACB combined with LIA around knee joint group (group AL). ACB was performed with 0.5% ropivacaine 15 ml after endotracheal intubation in group A and group AL, and in addition LIA was performed around the knee joint after the osteotomy was completed during surgery in group AL.The patient-controlled ACB analgesia was applied at the end of surgery in both groups.The analgesic solution contained ropivacaine 400 ml (in 0.9% normal saline 200 ml), and the analgesic pump was set up to deliver a 5 ml bolus dose with a 30-min lockout interval and background infusion at 5 ml/h.When visual analog scale score>4, and pain was still not relived at 30 min after pressing by patients, pethidine hydrochloride 100 mg was intramuscularly injected as rescue analgesic.Peripheral venous blood samples were collected immediately before surgery (T 0) and at 24, 48 and 72 h after surgery (T 1-3) for determination of serum interleukin-6 (IL-6) and IL-10 concentrations by enzyme-linked immunosorbent assay.The muscle strength on the affected side was assessed at T 1-3.The patients′ satisfaction score, requirement for rescue analgesia, and adverse effects were recorded. Results:Compare with group A, the serum IL-6 concentrations were significantly decreased and serum IL-10 concentrations were increased at each time point after surgery, postoperative patients′ satisfaction scores were increased, the requirement for rescue analgesia was decreased ( P<0.05), and no significant change was found in the quadriceps strength of the affected limb and incidence of adverse reactions after surgery in group AL ( P>0.05). Conclusion:ACB and LIA around the knee joint can mitigate postoperative inflammatory responses in the patients undergoing TKA.