In order to reinforce the development and sharing of elaborate educational resources and improve the educational quality, the course team completed the transformation and upgrading of the original fine course for the construction of national high-quality resource sharing class. In the con-struction of courses the organic combination of course content, method, skills, quality was focused on, and the transformation and upgrading of the course achieved further optimization of the excellent resources and full sharing.

Objective:To observe the influence of isehemia-reperfusion(I/R)on hepatoma growth and on the expression of genes associated with tumor metastasis and recurrence(VEGF and MMP-9)in the adjacent tissues of cancer in nude mice. Methods:BALB/c nude mouse model bearing Hep3B-tumor in the liver was established and the model mice were evenly randomly into 5 groups:sham group and ischemia/reperfusion 1 h,6 h,5 d,and 7 d groups(n=8).I/R models were established by blocking porta hepatic;the sham group underwent the same treatment as the I/R model group except for blocking of porta hepatic.ALT and AST were detected in I/R 1 h and 6 h groups.Real-time-PCR was employed to detect the change of VEGF and MMP-9 in the adjacent tissues of cancer and the results were compared with that of the control group(n=6). Histopathological changes of liver were studied by H-E staining and necrotic areas were calculated in I/R 5 d and 7 d groups (n=6).The remnant tumor bearing mice were sacrificed 2 weeks after I/R to measure the volume and mass of the tumors. Results:Two weeks later,the tumor volume and mass in I/R group were increased compared with those in the sham group ([209.6?25.74]mm~3 vs[330.6?32.01]mm~3,[0.214?0.036]g vs[0.374?0.045]g,P

Objective:To explore the change of serum interleukin 16 (IL-16) level and the effects of highly-active antiretroviral therapy (HAART) on IL-16 in patients with HIV infection.Methods:77 patients with HIV infection were studied,with fifteen normal subjects studied as controls.The patients were subdivided into stages according to the standards by US Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO).Of all the individuals,the CD4+T cells and CD8+T cells and the amout of IL-16 were measured at each stage to find the difference among normal and groups of the patests,and between the groups with and without HAART.Results:The level of CD4+ T cells in the experimental group were lower than that of the control group in of the patients (P

Objective: To explore the effect of high glucose on NADPH oxidase (NOX) expression and intracellular reactive oxygen species (ROS) generation in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were divided into a control group, a mannite group, a glucose group and aglucose plus diphenylene iodonium (DPI) group. Intracellular ROS was detected by lfow cytometry. RNA and protein expression of NOX in HUVECs was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. Results: 1) Compared with the control group, the intracellular ROS were signiifcantly increased in the glucose group (P<0.05,n=3), but there were no signiifcant change in the glucose plus DPI group (P>0.05,n=3); 2) Compared with the control group, the mRNA and protein expression of NOX4 in the glucose group were signiifcantly increased (P<0.05), but there were no signiifcant change in the mRNA and protein expression of NOX2, p22phox, p67phox and rac (allP>0.05); 3) there were no signiifcant difference in the mRNA and protein expression of NOX2, NOX4, p22phox, p67phox and rac between the glucose plus DPI group and the control group (allP>0.05). Conclusion: High glucose may increases intracellular ROS generation by increasing the expression of NOX4 in HUVECs, which might mediate the oxidative stress.

Objective Treatment and histopathological observation of demodex canis in beagle dogs .Methods Using the method of direct smear for microscopic examination of demodex canis.Histopathological observation on the skin of the parasitic parts after routine paraffin section and HE staining .Results (1) Clinical observation: The red spots and hair removal was appeared on limbs , eyes, lower abdomen and other parts of the skin of canine patients .The skin of the limbs becomes thicker and wrinkles .(2) Blood routine examination:Basically normal.(3) Microscope observation:The results showed that a large number of worms and eggs of small demodex canis could be found .(4) Histopathological observation:Hair follicles showed a large number of demodex mites and eggs.The sebaceous glands and sweat glands have normal morphology and no mites was found .A large number of eosinophils and neutrophil infiltration were seen around the hair follicles.It was also found that the formation of multifocal granuloma:the granuloma was oval shaped .(5) Treatment programme:The combination of medication and the strengthening of environmental control has been shown to be effective . Conclusions Granuloma caused by demodex canis can be divided into immune granuloma .It may not be possible to destroy the sebaceous glands after infection with small demodex canis.Whether the sebaceous gland is infected with the demodex canis may be associated with the worm species or course of disease .

The effects of epidermal growth factor (EGF) on proliferation of G15 glioma cells and the possible mechanisms were investigated. GFAP and EGFR expression was detected by immunohistochemical method. After the cells were treated with EGF at different concentrations, cell count method was used to determine the proliferation of glioma cells, cell cycle and apoptosis were analyzed by flow cytometry (FCM), and laser scan confocal microscope (LSCM) was used to measure the cytoplasmic free calcium. The results showed that GFAP was diffusedly expressed in GL15 cells and EGFR was over-expressed. EGF at doses of < or =1 ng/mL could significantly stimulate cell proliferation, cells in phase G0/G1 decreased, and those in phase S increased. EGF at doses of 10 and 100 ng/ml could inhibit the cell proliferation significantly, and the apoptosis ratio in high dose of EGF group was higher than in control group. EGF could significantly induce a quick rise of intracellular free calcium, but the peak value of intracellular free calcium activated by high dose of EGF was higher than by low dose of EGF. It was suggested that EGF had a dual effect on gliomas: low dose of EGF could stimulate the cell proliferation of gliomas, but high dose of EGF could induce the cell apoptosis and inhibit the proliferation of gliomas, which might be contributed to the difference of intracellular free calcium.

This study examined the effect of GHRP-6, a known GHSs receptor agonist, on the phosphorylation of cAMP-responsive element-binding protein (CREB) and the underly mechanism. GH3 cells were cultured and subjected to different treatments as follows: GHRP-6, GHRP-6 plus GHRH, phorbol ester (PMA), an activator of PKC, alone or in combination with GHRP-6, Gö6983, a general inhibitor of PKCs, in the presence or absence of GHRP-6, rottlerin, an inhibitor of PKCs, alone or plus GHRP-6. The cells were transiently transfected with PKCsigma-specific siRNA and then treated with GHRP-6. GH level was measured by enzyme-linked immunosorbent assay (ELISA). The expression of phosphor-CREB, PKCsigma, PKCtheta and phosphor-PKCsigma was determined by Western blotting. The results showed that GHRP-6 stimulated GH secretion in both time- and dose-dependent manners and enhanced the effect of GHRH on GH secretion. GHRP-6 was also found to induce CREB phosphorylation. Moreover, GH secretion was enhanced by the PKC activator PMA and reduced by the PKC inhibitors (Gö6983, rottlerin) and knockdown of PKCsigma. PKCsigma could be activated by GHRP-6. It is concluded that PKC, especially PKCsigma, mediates CREB phosphorylation and GHRP-6-induced GH secretion.

Aim To explore the anti-apoptotic function of cardiac progenitor cells(CPCs)-derived exosome in vitro.Method CPCs were isolated from mouse heart using Magnetic Cell Sorting(MACS)system.Flow Cy-tometry(FC)determine the purity of stem cell surface antigen-1 positive(Sca-1 +)CPCs.Exosome was puri-fied from conditional medium,and confirmed by West-ern blot using CD63 as a marker,Nanoparticle Traffic-king Analysis(NTA)was used to detect the diameters and concentration of exosome.Then the cells were di-vided into control groups and CPC-exosome pre-protec-tion groups.H2 O2 was added into H9c2 cells to induce oxidative stress.Western blot was adopted to determine the expression of cleaved caspase-3.Results ① Im-munofluorescence showed that CPCs isolated by MACS were positively expressing Sca-1 protein;FC analysis showed that typical purity of Sca-1 +CPCs from the first
preparations was more than 95%.② WB demonstrated that CD63 of exosome isolated from CCMwas positively expressed,and NTA results showed that the diameters of exosome were (82.33 ±3.06)nm(n =3).Micro-scope detected PKH-26 labeled exosome appeared in the cytoplasma of H9c2 cells.③ Western blot showed the CPC-exosome pre-protection groups significantly down-regulated the levels of cleaved caspase-3 com-pared to the control groups(P <0.05).Conclusion CPC can secrete exosome which carries many important cargos,which can effectively gather in H9c2 cells. CPC-exosome can protect H9c2 cells from the oxidative stress induced by H2 O2 .Our results highlight a new perspective strategy for cardiac disease.

Objective To establish a mouse ’ s cross-territory ear flap that enables chronic , in-vivo observation of the change of vascular morphology .Methods 30 ICR mice, weighing 25~40 g, were used for this study .Commercial depilatory cream was used to first remove the hair of the mice , after which the vascular pattern in the ears was investigated . According to the observation of the vascular pattern in the mouse ’ s ear, the eye scissors were used the sever the outer 2/3 of the base of the ear , in which process a ear ’ s flap based on a vascular pedicle but crossed three vascular territories was created.After the creation of the flap , the mice were placed on an automatic controlled movable machine with the ear ’ s flaps spread over a customized Plexiglas .Then the flaps were photographed under the stereoscope ( ×25) at the following time points:1,2,3,5,7,10,14,21,30 d.the necrosis of the flap, and the morphological change of the vessels within theflap were analyzed .Results The ICR mouse ’ s ear was supplied three angiosomes , which were respectively named as the cephalic , median and caudal angiosomes from inside out .Five days after the flap’s creating, necrotic rate of(15 ±7)%was developed .The choke vessels between the medial and median angiosomes expanded rapidly in diameter , reaching the plateau 10d after flap creation, resulting the dilated choke veins and arteries at their peak being 3.9 ±0.5 and 3.5 ±0.7, respectively, than their initial sizes.The diameter of the choke veins began to shrink at approximately 10d, stabilizing after 21d.The diameter of the choke arteries plateaued and stabilized at around 10d.Conclusion ①after harvest of extended flap, the dilation of veins seemed to passive , whereas the dilation of arteries seemed to active;②the number of the choke vessels between the dynamic and potential territories that are involved in dilation and extent of the dilation are much smaller than that of the choke vessels between the anatomic and dynamic territories;③the mouse ’ s ear flap is an excellent model of further study of mechanism underlining the dilation of choke vessels and for the screening of vasoactive drugs that augment the survival of the large flap .

Objective To provide the anatomical basis for the flap based on the perforator of Plantar arch,through investigation of the morphological features of the perforator of the arch of the foot.Methods From November,2015 to March,2016,the first metatarsal base and the fifth metatarsal tuberosity were chosen as the observation point on 25 specimens of adult human feet perfused with red latex.The following contents were observed under surgical magnifier:①The origin,courses,branches and distribution of the perforator of Plantar arch.②The anastomoses among the perforator of Plantar arch and the fete arteriosum dorsale pedis.Mimic operation was performed on another fresh specimen perfused with red latex.Results There were 3 perforators in Plantar arch,which passed through the 2nd-4th metatarsal dorsal muscles to the dorsi pedis and then divided into an ascending branch and a descending branch.The ascending branch anastomosed with the rete arteriosum dorsale pedis,and the descending branch stretched to the 2nd-4th plantar arteries.The initiative outer diameters of the 1st-3rd dorsal perforators of Plantar arch were (1.5 ± 0.3)mm,(1.1 ± 0.4) mm and (0.9-± 0.3) mm respectively,and the lengths of the stem were (1.1 ± 0.2) cm,(1.5 ± 0.1) cm and (1.5 ± 0.5) cm respectively.Conclusion The flap can be used for repair of soft-tissue defects of dorsal and front foot through dorsal transposition or a V-Y advancing flap with the perforator of Plantar arch as its vascular pedicle.