Objective To report the clinical effect of repair of significant tongue defect after tongue cancer radical resection using anterolateral thigh perforator free flap with CTA three-dimensional reconstruction technique assistance.Methods From January,2012 to November,2015,13 cases were performed using anterolateral thigh perforator free flap in the repair of significant tongue defect after tongue cancer radical resection.Preoperation CT scan of the free flap donor site was done to obtain the three-dimensional images of arterial blood area by Digital Three-dimensional Reconstruction CT Technique to determine the origin of the anterolateral thigh flaps,direction,classification,length,diameter and the position of pedicle perforator.According to the condition of the defect of the tongue,flap of area from10 cm×9 cm to 8 cm×6 cm was design,the flap arteries and veins were anastomosed with the external maxillary artery or the superior thyroid artery,anterior vein,internal jugular vein or external jugular vein respectively.The shape and recovery of function of the reconstructed tongue were observed regularly after operation.Results All the flaps in the 13 cases survived,in the postoperative 4 to 18 months followed up,the appearance of the reconstructed tongue was close to the normal one,the patients were satisfied with the mastication and feeding function.Conclusion CTA three-dimensional reconstruction technique assisted anterolateral thigh perforator free flap technique is useful in obtaining good clinical effect,and it is one of the most best methods for repairing significant tongue defect after tongue cancer radical resection.

Objective To establish a novel glucocorticoid (GC)-resistant human diffuse large B lymphoma(DLBCL) cell line Toledo/dexamethasone (DEX) by the exposure to DEX,and observe the biological characteristics of resistant and parental cell line,investigate the mechanisms of glucocorticoid-resistance.Methods Toledo/DEX was established by the exposure to DEX,the dose of which was increased gradually and intermittently for long periods of time.Toledo,in the logarithmic growth phage,was incubated in the culture medium containing DEX at the concentration of 1×10-8 mol/L at first.The medium without DEX was replaced after for 96 hours until the cell line re-entered the logarithmic growth phase.Repeat the above steps to acquire the ultimate concentration of DEX in the medium as 1.024 ×10-5 mol/L.The biological characteristics of resistant and parental cell lines were evaluated.Results Toledo/DEX was more invasive in the aspects of ultrastructure,tumorigenicity and drug sensitivity.Meanwhile,Toledo/DEX achieved some stable biological characteristics such as morphology,karyotypes and immunophenotype.Furthermore,GC receptor (GR) α and GR β protein expression analysis showed that GR was involved in the mechanism of the GCresistance.Conclusions Toledo/DEX is a drug-resistant cell line with a stable biology backgroud.These results may help shed light on the knowledge of GC-resistance and lay the groundwork for searching new therapeutics to reverse drug-resistance.

Aim To investigate the protective effect and mechanism of madecassoside from hydrocotyle sibthorpioides (MHS)on learning and memory impair-ment induced by D-galactose (D-gal)in mice.Meth-ods Totally 75 SPF Kunming male mice were divided into normal control group,model group,low-,middle-and high-doses of MHS treated groups.The dementia models were induced with D-gal.The learning and memory functions were tested by Morris water maze, the level of Aβ1 -42 and its related proteins in the hip-pocampus was determined by Western blot,and the ex-pression of Aβ-related genes were determined by RT-PCR.Results Compared with model group,MHS markedly decreased the content of Aβ1 -42 ,inhibited the expression of APP,BACE1 and CatB,but promo-ted the expression of NEP and IDE.In addition,AHS significantly increased the expression of plasticity-relat-ed proteins including PSD-95,p-NMDAR1,p-CaMKII, p-PKACβ,PKCγ,p-CREB and BDNF.Conclusions MHS could remarkably ameliorate the learning and memory impairment induced by D-gal in mice,which may be due to its ability to inhibit the Aβgeneration and deposition and promote synaptic plasticity related protein expression.

Objective: To study the attenuating effect of angiotensin I type 1 receptor antagonist losartan and an-giotensin converting enzyme inhibitor captopril on aortic atherosclerosis in rabbits. Methods: Thirty-one male New Zealand rabbits were randomly divided into 5 groups:control group,high cholesterol diet group,losartan group, captopril group and combined drug administration groupdosartan+captopril). The animals were killed after 16 weeks and the serum total cholesterol ,triglyceride, high and low density cholesterol .atherosclertic ratio,endothelin,NO,plaque area percentage,aortic cholesterol content and vascular smooth muscle cell (VSMC) apoptosis were determined. Results:The plaque area percentage,aortic cholesterol contents and endothelin levels of 3 drug treatment groups were significantly lower than that of high cholesterol group,NO contents and VSMC apoptosis were significantly higher. Conclusion:Losartan and captopril can attenuate aortic atherosclerosis induced by high cholesterol diet .combined administration of the 2 drugs at low doses are more effective. The mechanism may be related to the protection of endothelial function and the effect on apoptosis of VSMC.

Objective To study the effect of Hydrogen on ischemia/reperfusion(I/R)-induced cardiocyte apoptosis and apoptosis related proteins expression in diabetic rats .Methods Diabetes mellitus was induced by intraperitoneal injection of streptozotocin (STZ) ,then feed 4 weeks before build ischemia/reperfusion model .60 SD male rats ,weight during 300-350 g ,were randomly divid-ed to six groups :non-diabetic sham-operated group (NS) ,non-diabetic I/R group(NI/R) ,non-diabetic hydrogen treated group (NH) ,diabetic sham-operated group(DS) ,diabetic I/R group(DI/R)and diabetic hydrogen treated group(DH) ,each group has 10 rats .The cardiac muscle I/R model was made by 30 min occlusion of the left anterior descending coronary artery and 2 h reperfu-sion .The rats in Hydrogen group were treated with 5 mL/kg hydrogen by intraperitoneal administration at the beginning of reper-fusion .The apoptosis index (AI) was calculated by TUNEL .The positive expressions of Bcl-2 ,Bax ,Caspase-3 in cardiomyocytes were respectively detected by immunohistochemistry .Results The apoptotic rates of cardiomyocytes and the positive expressions of Bcl-2 ,Bax ,Caspase-3 were significantly increased(P<0 .01) ,Compared with I/R group ,the apoptotic rates of cardiomyocytes in hydrogen treatment group were obviously decreased(P<0 .01) ,and the positive expressions of Bcl-2 were increased(P<0 .01) ,at the same time ,the positive expressions of Bax ,Caspase-3 were decreased(P<0 .01) .Conclusion Hydrogen inject by intraperitoneal method on myocardial ischemia-reperfusion injury of diabetic rats has a protective effect ;Its mechanism may be related to its inhibi-ting myocardial apoptosis by advanced the Bcl-2 protein expression and reduced the Bax 、Caspase-3 protein expression .

Objective To establish a nude mice model for NB4/ShHMGA2 xenograft and explore the effect of HMGA2 knockdown on hematological malignancies. Methods NB4/ShHMGA2 or NB4/ShControl cell lines were established by transfecting the recombinant Lentivirus-HMGA2shRNA and the vacant Lentivirus-NC-marked into NB4 cells. The knockdown of HMGA2 was identified by RT-PCR and Western blot. Ten male BALB/c nude mice aged 4 ~ 5 weeks were equally divided into two groups. The mice irradiated by 4 Gy 60 Co were subcutaneously injected with 8 × 106 NB4/ShHMGA2 or NB4/ShControl cells into one side of axilla. The volumes of xenograft tumor were evaluated using the equation volume (mm3) = (L × W2)/2. The xenograft tumor section was detected by IHC with Ki-67 antibody. Results NB4 cell xenograft tumors developed in all mice of both the two groups. The NB4/ShHMGA2 cells in the nude mice grew at a lower rate than those in the controls. There were statistically significant differences in the volume and weight of xenograft tumor between the two groups [(1 484.25 ± 156.342)mm3 vs (3 228.674 ± 285.64)mm3, P < 0.05] and [(2 135.33 ± 198.05) mg vs (650.46 ± 85.12)mg, P < 0.05]. The Ki-67 protein level in NB4/ShHMGA2 cells xenografts was lower than that in the controls. Conclusion The knockdown of HMGA2 could inhibit proliferation of NB4 cells in NB4 cells xenograft tumor.

Objective: To study the chemical constituents of Changyanning Tablets, a high performance liquid chromatography-quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS/MS) was established to recognize and classify the ingredients accurately and rapidly. Methods: Agilent ZORBAX Eclipse XDB-C18 chromatographic column (250 mm × 4.6 mm, 5 μm) was employed and the separation was performed with the mobile phase consisting of methanol-0.05% acetic acid aqueous solution. The information of accurate mass and multistage fragment ions were obtained by the monitored simultaneously for positive and negative ions. The main chemical constituents of Changyanning Tablets were identified by high resolution mass spectrometry data, combining with Pubmed, Hmdb, Massbank network database, reference literature and comparing the reference. Results: Fifty-one chemical components were finally identified in this study, including two phenylpropanoids, eight iridoids, 12 flavonoids, four tannins, 23 organic acids, and two other classes. Conclusion: This study comprehensively studies the material basis in Changyanning Tablets, which provides a basis for improving the quality evaluation system of Changyanning Tablets and lays the foundation for elucidating the active components mechanism.

We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.

[ ABSTRACT] AIM:To observe the effect of high glucose on the protein expression of calreticulin ( CRT) and its association with cell apoptosis and mitochondrial dysfunction in the cardiomyocytes.METHODS: AC-16 cardiomyocytes were randomly divided into normal glucose group, high glucose group, high glucose+CRT siRNA group and isotonic con-trol group.The cell apoptotic rate, reactive oxygen species (ROS), mitochondrial membrane potential level, respiratory enzyme activity, and protein expression of CRT were observed.RESULTS: Compared with the cardiomyocytes in normal glucose group, the apoptotic rate and ROS production of cardiomyocytes increased in high glucose group, accompanying with the decreases in the mitochondrial membrane potential level and enzyme activitiy of the respiratory chain.The protein expression of CRT was significantly increased in high glucose group.However, compared with high glucose group, high glucose+CRT siRNA decreased the expression of CRT and attenuated the damage of mitochondria, but CRT siRNA did not reduce the ROS level in cardiomyocytes.CONCLUSION:High glucose brings about CRT over-expression to induce mito-chondrial injury, thus increasing myocardial apoptosis.

Objective To explore the clinical value of applying digital technology to make the course of toe vein clear before toe free flap transplantation for thumb reconstruction.Methods Before operation,scan the donor and recipient site by CT,and apply digital three-dimensional CT angiography reconstruction technology to got threedimensional images of the arteries and veins in the donor site for 22 various defected thumbs which were going to have toe free flap transplantation operation,thus indentifyed the types of FDMA and the distribution and course of the toe vein from June,2012 to March,2013.Used computer assisted anatomical modeling to analyze three-dimensional images of blood supply of the donor site and degree of defect of the recipient site and guide the operative design in the donor site according to the digital three-dimensional reconstruction technology.Conducted toe free flap transplantation for thumb reconstruction in accordance with the design.Results It shows that 64-slice dual-source spiral CT could well display the size,distribution and course of toe vein.The toe veins were clarified to two groups,deep one and superficial one whose diameter was larger and was the main reflux vein of the toe.Dorsal vein of the toe origind from venous plexus in the nail bed and it travels along the back of the toe to reach to the near side of metatarsophalangeal joint.And then it beeomes three to four dorsal metatarsal veins to infused dorsal venous arch of foot.The rule of its course was:centralize-decentralize-connect-decentralize again.The great saphenous vein origins from the inner side of dorsal venous arch of foot,rises along the crus before malleolus medialis and collect three larger tributaries:medial malleolar branches-anterior crural branch-outside crural branch.The small saphenous vein derives from the outside of dorsal venous arch of foot and winds behind lateral malleolus up to the back of the curs and infuses to the vein.Conclusion It is useful for venous return,reducing the incidence of vascular crisis by applying toe flap to reconstruct thumbs and clarifying the course and distribution of toe vein with the aid of digital three-dimensional technology to design operation,thus improving the survival rate of toe flap transplantation.