Objective To investigate the care measures of microspheres arterial embolization in the treatment of primary liver cancer after TACE.Methods The chemotherapy drugs were injected through the catheter superselectively inserted to the left and right hepatic arteries,microspheres and super liquefied iodipin were used for tumor embolization,preoperative and postoperative care were provided to patients.Results Through the full nursing care of 50 cases of liver cancer patients,the occurrence of adverse effects and complications after surgery were reduced,enthusiasm of patients with treatment were confirmed,and the effect of the treatment was improved.Conclusions Treatment of liver cancer by hepatic artery perfusion chemo-embolization is an effective means,using scientific methods of effective care for patients is very important.

Purpose:To detect the expression of a novel i nh ibitor gene of apoptosis,survivin,in gastric cancer and in gastric carcinoma MGC -803 cell line,also to analyze its correlation with the expression of COX-2. Methods:In 93 stomach carcinoma tissues and 20 normal gastric tissues , the expression o f survivin and COX-2 were examined by using the streptavidin-biotin peroxidase (SP) method. Results:In contrast to negative expression in normal gastric mucosa,survivin was express ed in 66 of 93(71%) cases of gastric cancer samples,Overexpression of survivin i n gastric carcinoma MGC-803 cell line was also found,There was a relationship b etween survivin gene expression and degrees of differentiation,lymph node metast ases and TNM stages.The expression of survivin was positively correlated with th at of COX-2(liner index of Pearson=0.227 P

BACKROUND:Corneal hemangiogenesis occurs in 40%-60%patients after keratoplasty.Blood vessel is one of the high risk factors for corneal immunological rejection.To inhibit corneal hemangiogenesis would prolong the survival time of the grafts and promote the successful rate of the keratoplasty.OBJECTIVE:To explore the inhibitive effects of doxycycline on corneal angiogenesis after keratoplasty.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the State Key Laboratory of Ophthalmology(No.2006DA105054),Zhongshan Ophthalmic Center,Sun Yat-sen University from March to August 2007.MATERIALS:A total of 48 healthy dean Sprague Dawley rats served as recipients(right eye)and 24 Wistar rats as donors(both eyes).CD31-PEfluorescent antibody was obtained from Sigma,USA.Sandwich enzyme-linked immunosorbent assay(ELISA)kit for vascular endothelial growth factor(VEGF)was brought from RapidBio,USA.METHODS:Corneal allogenic transplantation models were established in rats.Recipients were equally and randomly divided into 2 groups:saline control group and doxycycline group.Twenty minutes prior to surgery,mydriasis was performed using 1%atropine,with a diameter of 2.75 mm of implant and 2.5 mm of implant bed.In the saline control group,conjunctiva of the right eye received saline,three times a day,following surgery.In the doxycycline group,conjunctiva of the right eye received 1%doxycycline,three times a day,till 30 days following surgery.MAIN OUTCOME MEASURES:The following parameters were measured:corneal angiogenesis using immunofluorescence,expression of VEGF protein by using ELISA.RESULTS:Compared with the survival time of saline control group[(9.67±2.73)days],the mean survival time of doxycycline group[(20.67±3.01)days]was significantly prolonged(P＜0.01).The mean percentages of neovascularized corneal area in the saline control group were(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%at 3,7 and 14 days after keratoplasty,respectively.The mean percentages of neovascularized corneal area in the doxycycline group were(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%at the same time point respectively.Compared with the saline control group,the mean percentages of neovascularized corneal area of the doxycycline group was significantly reduced at 7 and 14 days after keratoplasty(P ＜0.05).The expression of VEGF in the saline control group was(541.00±75.44)pg/mg,(960.00±90.14)pg/mg,(976.00±130.41)pg/mg at 3,7 and 14 days after keratoplasty,respectively,while expression of VEGF in the doxycycline group was(115.33±9.29)pg/mg,(239.00±41.62)pg/mg,(361.00±65.20)pg/mg,respectively.The difference of VEGF expression at all time points between the two groups was significant (P＜0.05).CONCLUSION:Doxycycline has a significant effect in inhibiting corneal angiogenesis and prolonging survival time of implants after keratoplasty.

BACKGROUND: Cervical lymph nodes are draining region of cornea. It is believed that aqueous fluid goes through a minor pathway named uveoscleral drainage, which will allow passage of antigen-presenting cells (APC) directly to the draining lymph nodes and induce allograft rejection after keratoplasty.OBJECTIVE: To explore the inhibitory effects of cervical lymphadenectomy in alkali induced high-risk corneal transplantation.DESIGN: A randomized controlled animal experiment.SETTING: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: The experiment was performed in the State Key Laboratory of Ophthalmology (No. 2006DA105054), Zhongshan Ophthalmic Center, Sun Yat-sen University from May 2005 to February 2007. 144 male animals (1-2 months old) including 104 SD rats and 40 Wistar rats were provided by the animal experimental center of Sun Yat-sen University. Sandwich enzyme-linked immunosorbent assay (ELISA) kits for interleukin-2 (IL-2) and interferon-γ (IFN-γ) were brought from BioSource International company (USA). The animal treatment in the experiment was accorded with the statement in Association for Research in Vision and Ophthalmology (ARVO) for animals.METHODS: With the SD rats as recipients, and Wistar rats as donors, all rats were subjected to corneal allografting. The recipient rats were randomly divided into 4 groups (n=20): group A (control group) which underwent corneal transplantation; group B which was subjected to bilateral cervical lymphadenectomy; group C, corneal transplantation 21 days after the alkali burn injury; group D, cervical lymphadenectomy following group C. The immune rejection of grafts was evaluated by detecting the expression of IFN-γ and IL-2 using ELISA. The time when allograft rejection occurred was recorded and mean survival time (MST) was compared among the groups. The development of corneal inflammation and new vessels was examined by slit lamp microscope and histopathological examination.MAIN OUTCOME MEASURES: ①The development of corneal inflammation after corneal alkaline burns. ②MST of rats in each group following transplant. ③The expression of IL-2 and IFN-γ in grafts of each group. RESULTS: ①Normal rat cornea was transparent without inflammation or neovascularization. There were many inflammatory cells invading to stroma on day 3 after burn. Then, the inflammation of cornea resolved gradually 3 weeks after the burn, but corneal neovascularization reached the peak at that time. Corneal blood vessels regressed completely at the end of 8 weeks after the burn. ②The MST of group A, B, C, and D was (10.40±1.14), (46.30±9.46), (7.00±1.58), and (15.00±3.39) days, respectively. Compared with the group A, the MST of group B was significantly longer (P < 0.05), and the MST of grafts in group D was also significantly longer than group C (P < 0.05). ③The expression of IFN-γ and IL-2 proteins was absent in group B. Compared with group C, the expression of IL-2 and IFN-γ proteins in group D significantly decreased on days 3, 7, 10, and 14 after keratoplasty (P < 0.05). CONCLUSION: Cervical lymphadenectomy therapy can effectively inhibit corneal allograft rejection in normal and high-risk corneal beds after alkali burn injury.

Objective To observe the effects of Tanshinone Ⅱ A sodium sulfonate (TSS ) on ischemia/reperfusion (I /R) induced cardiac injury in male (Sprague-Dawley,SD ) and explore its mechanisms.Methods Rats were subjected to a 30 min coronary arterial occlusion followed by 24 hours reperfusion.The survival rats were randomly (random number)divided into sham group (Sham group,n =10),ischemia reperfusion group (I /R group,n =10),low dose of TSS group (TSS-L group,n =10), medium dose of TSS group (TSS-Mgroup,n =9),high dose of TSS group (TSS-H group,n =9).A MAP heart function analysis system was used to measure hemodynamic variables,and TTC staining method was used to detect the myocardial infarct size.The levels of Bcl-2,Bax,Caspase-3,Lc3B/Lc3A,Beclin-1 and high mobility group box1 (HMGB1)were detected by western blot method.All data were analyzed by using One-way analysis of variance (ANOVA)(LSD-t test).Results Cardiac function in I /R group was lower than that in Sham group,and that was significantly improved by pretreated with TSS (P <0.05),but there were no significant differences in Pmin and DAP between Sham group and I /R group (P >0.05 ).The percentage of myocardial infarct size in TSS pretreatment group was significantly smaller than that in I /R group (P <0.05 ).Compared with Sham group,levels of Caspase-3 and Bax increased,and the Bcl-2 content was reduced obviously in I /R group (P <0.05).TSS pretreatment significantly down-regulated the levels of Caspase-3 and Bax protein (P <0.01).At the same time,the level of Bcl-2 was increased in all TSS pretreatment groups (P <0.01).Compared with Sham group,the ratio of Bcl-2 /Bax in I /R group was lower (P <0.05),and that was elevated in TSS groups (P <0.05 ).The change of autophagy related protein beclin-1 and Lc3B/Lc3A was in similar trend,and the levels of beclin-1 and Lc3B/Lc3A in I /R group were lower than that in Sham group (P <0.05),and those were raised in TSS pretreatment groups (P<0.05).The level of HMGB1 in I /R group was higher than that in Sham group (P <0.05),and compared with I /R group,the level of HMGB1 significantly decreased in TSS pretreatment groups (P <0.01 ). Conclusions The tanshinone ⅡA sodium sulfonate can protect the myocardium from ischemia/reperfusion injury and the mechanism may be attributed to the inhibition of cell apoptosis and activation of cell autophagy.

BACKGROUND:With the deepen understanding on the biological function of Rho/ROCK pathway, new ROCK inhibitors continue to be discovered, and ROCK inhibitors show good promoting effects on the survival, proliferation and migration of keratocytes. Research on ROCK inhibitors wil provide more donor materials or seed cels for regenerative medicine and clinical cel transplantation. OBJECTIVE:To summarize and explore the progress in the treatment and application of corneal disease using the ROCK inhibitors Y-27632 and Y-39983. METHODS:The PubMed database and CNKI database were retrieved by computer to search the relevant literature published between 2008 to 2015 using the key words of “corneal endothelial cel, corneal epithelial cel, ROCK inhibitor, Y-39983, Y-27632” in English and Chinese, respectively. Relevant articles in line with the theme were screened and analyzed. RESULTS AND CONCLUSION:Totaly 264 papers were initialy searched. At last, 45 papers were selected. Currently there are two main ROCK inhibitors: Y-27632 and Y-39983, but both of which are stil in basic research stage and clinical testing stage. Y-27632 promotes the proliferation and activity of corneal epithelial stem cel after resuscitation; Y-39983 as a novel ROCK inhibitor can be better to inhibit Rho kinases activity than Y-27632, thereby more effectively promoting the healing of the corneal endothelium. There are many studies on the application of ROCK inhibitors in corneal treatment, but not a stable method established to obtain seed cels. Each method has its own advantages and disadvantages, and how to overcome these disadvantages and to find fast and stable access to seed cels is the future direction of development.

Objective To explore the clinical characteristics of epidural hematoma in children. Methods A total of 120 children with epidural hematomas within recent three years were reviewed. Results The main cause of injury in infants and preschool children was falling or sliping, but traffic accident was the predominant cause in children over seven years old. About 65.8% children were complicated by skull fractures, with average Glasgow Coma Scale (GCS) score of 13.6. Except for acute hematoma treated with emergency surgical operation, the other hematoma was rechecked with CT scan at days 1 and 3 or so after it was found for the first time. Patients receiving operation accounted for 57.5% and those with hematoma due to diploe bleeding for 43.9%. Conclusions The primary cerebral injury is not severe relatively in children with epidural hematoma, in which the incidence of skull fracture is lower than that in adults. The main cause for hematoma formation is diploe bleeding. Sound prognosis can be obtained through recheck of CT scan and suitable therapy.

Objective:To examine the correlations of microRNA-34c(miR-34c) expression in the peripheral blood with the onset of diabetic foot ulcer(DFU)and diabetic foot osteomyelitis(DFO)in patients with type 2 diabetes mellitus(T2DM).Methods:Sixty newly-diagnosed T2DM patients without DFU(T2DM group), 112 T2DM patients with DFU(DFU group), and 60 healthy controls with normal glucose tolerance(NC group)were included. The 112 T2DM patients with DFU were further divided into DFO( n=64)and NDFO( n=48)groups. The levels of miR-34c were determined by quantitative real-time PCR, while clinical features and risk factors of DFU and DFO were explored. Results:A significant increase in the expression level of miR-34c in peripheral blood was observed in T2DM group compared with NC group[2.99(1.45-6.22) vs 1.01(0.89-1.52), P<0.05], and a markedly increased miR-34c expression level was noted in DFU group compared with T2DM group [9.65(6.15-18.63) vs 2.99(1.45-6.22), P<0.01]. Additionally, the expression level of miR-34c in peripheral blood significantly increased in DFO group compared to NDFO group [13.46(8.89-19.11) vs 6.02(5.93-14.72), P<0.01]. Moreover, there was a positive correlation between the expression level of miR-34c in peripheral blood and the amputation rate in patients in DFU group( P=0.030), and a negative correlation in the expression level of miR-34c( P=0.025)with healing rate of DFU after eight weeks. The multivariate logistic regression analysis confirmed that a high expression of miR-34c was an independent risk factor for DFU and DFO( OR=3.52, 4.13; both P<0.01). Conclusion:An increased expression of miR-34c in peripheral blood of T2DM patients might be closely related to the occurrence, development, and prognosis of DFU and DFO.

Objective To construct a recombinant adeno-associated virus(rAAV)vector containing a human anti-epidermal growth factor receptor(anti-EGFR)single-chain variable fragment antibody gene,and observe its inhibitory effects on pancreatic cancer cell lines.Methods Human anti-EGFR single-chain variable fragment antibody gene was inserted into the Kpn I and Bgl Ⅱ sites to construct a rAAV-anti EGFR vector,and then rAAV1-EGFP group and rAAV1-anti EGFR group were established.The expression of anti-EGFR antibody was observed.Antibody expression was detected by Western blot,and the inhibition and apoptosis rates of human pancreatic cancer cell lines(PCT-3,SW1990,Capan-1,ASPC-1,MiaPaCa-2 and PANC-1 cells)were detected by CCK-8 assay and flow cytometry,respectively.All data were analyzed using the t test.Results The results of Western blot assay demonstrated that anti-EGFR antibody was expressed in 6 pancreatic cancer cell lines.The inhibition rates of rAAV1-EGFP and rAAVl-anti EGFR on pancreatic ASPC-1 cells were 1.1%± 2.4% and 15.1%±3.5%,respectively,with a significant difference between the 2 groups(t =6.598,P ＜0.05).The apoptosis rates of PANC-1 cells were 7.0% ± 3.0% in the rAAV1-EGFP group and 1 1.4% ± 2.5% in the rAAV1-anti EGFR group,with no significant difference between the 2 grouvs(t = 1.952,P ＞0.05).The apoptosis rates of SW1990,ASPC-1,Capan-1,PCT-3,MiaPaCa-2 cells were 1.1% ± 0.8%,1.5% ± 0.7%,1.7% ± 1.2%,1.1%±0.7% and 2.2% ± 1.1% in the rAAV1-EGFP group,and 17.6% ± 2.2%,46.9% ± 3.9%,20.0% ±2.8%,12.1% ± 1.6% and 31.1% ±2.5% in the rAAV1-anti EGFR group,respectively,with significant differences between the 2 groups(t = 12.208,19.846,10.405,10.909,18.327,P ＜0.05).Conclusions A rAAV-anti EGFR vector with human anti-EGFR single-chain variable fragment antibody gene was constructed.Anti-EGFR antibody has obvious inhibition effects on pancreatic cancer cell lines.

Objective EGFR targeted therapy mediated by adeno-associated virus is a promising way to treat pancreatic cancer.This study aimed to assess the feasibility and activity of combining rAAV-anti EGFR,gemcitabine,and radiation in pancreatic cancer cells.Methods Aspc-1 human pancreatic carcinoma cells were divided into several groups,in vitro and in vivo,which were respectively exposed to gemcitabine alone,radiation alone,rAAV-anti EGFR alone,the combination of rAAV-anti EGFR with gemcitabine,the combination of rAAV-anti EGFR with radiation,and the combination of all three agents.The pancreatic cancer tumor growth and apoptotic rate were measured.Results The apoptotic rate was higher in cells treated with a single or combination of agents compared to the negative control (P＜0.05).The combination of rAAV-EGFR,gemcitabine,and radiation produced the highest induction of apoptosis compared to a single agent alone (P ＜ 0.05).Treatment with rAAV-anti EGFR greatly inhibited growth in the tumor xenografts (P＜0.05),and a synergistic effect of rAAV-anti EGFR,gemcitabine,and radiation was found.The number of tissue cancer cells that expressed cleaved caspase-3 after treatment with rAAV EGFR was more than that of the control group (P＜0.05).The combined treatment of rAAV-anti EGFR,gemcitabine,and radiation induced the highest numbers of cells expressing cleaved caspase-3 compared to that with a single agent alone (P＜0.05).Conclusions The rAAV-anti EGFR therapy in combination with chemotherapy and radiation therapy demonstrated a greater efficacy over therapy with a single agent alone.rAAV-anti EGFR increased the efficacy of gemcitabine and radiation in the treatment of pancreatic cancer cells.