Objective To investigate the correlation between methylation status of UNC 5C gene promoter with colorectal cancer UNC5C .Methods 54 cases of sporadic colorectal cancer and related normal mucosal tissue ,as well as 6 colon cancer cell lines and fiber cell lines(FCL) in the oncology department of the Kailuan General Hospital from February 2010 to March 2013 were selected as the research objects and performed the mRNA and methylation analysis for exploring the correlation between the netrin‐1 receptor UNC5C defects with colorectal cancer disease .Results mRNA of UNC5A and UNC5B was expressed in the detected colorectal cancer cell lines .Except FCL for UNC5C ,all of the cancer cell lines had no mRNA expression .In colorectal cancer tissue , UNC5C methylation was significantly higher than that in the normal mucosa ,the difference was statistically significant (P<0 .05) . The methylation levels of UNC5C gene in the stage Ⅰ to Ⅱ ,and the stage Ⅲ to Ⅳwere significantly higher than non‐methylation levels ,the differences were statistically significant (P<0 .05) .The correlation analysis by the Spearman method showed that the UNC5C defects was positively correlated with colorectal cancer disease (r= 0 .856 ,P< 0 .05) .Conclusion Netrin‐1 receptor UNC5C defect has certain correlation with colorectal cancer disease .

Objective To investigate the effect of Yunnan Baiyao Plaster on serum related markers levels in the patients with stroke ,to evaluate its improvement effect on the neurological dysfunction and pain and to analyze its effect mechanism .Methods Eighty cases of hemorrhagic stroke treated in the neurosurgical department of our hospital from February 2015 to June 2016 were screened as the research subjects and divided into the control group and observation group according to the random number table method ,40 cases in each group .All subjects were given the routine treatment ,while the observation group received the external use of Yunnan Baiyao Plaster around the head operative incision and drilling ,to give Yunnan white ointment ,with the attaching area of 5 cm × 4 cm ,once daily ,for consecutive 7 d ,meanwhile took oral Yunnan Baiyao Plaster 0 .5 g with warm boiling water ,once daily for consecutive 7 d .The serum markers levels before and after treatment ,neural function defect score ,decline proportion of NIHSS scores and prognostic indicators were compared between the two groups .Results The mean pain value within 1 week in the obser-vation group was lower than that in the control group ,the difference was statistically significant (P<0 .05);the pain peak value and valley value within 1 weeks had no statistical difference between the observation group and control group ,the difference was not statistically significant (P> 0 .05);after 2 weeks ,ET ,TNF-α and IL-1β levels in the observation group and control group were lower than those before treatment in the intra-group comparison ,while the comparison between the two groups showed that the ob-servation group was lower than the control group ,the difference was statistically significant (P<0 .05);the decrease rate of NIH-SS scores within 2 weeks in the observation group was (74 .4 ± 5 .2)% ,which was higher than (66 .3 ± 10 .3)% in the control group ,the difference was statistically significant (P<0 .05);the occurrence rate of severe complications within 2 weeks in the ob-servation group was 7 .5% (3/40) ,the mortality rate was 2 .5% (1/40) ,which in the control group were 12 .5% (5/40) and 2 .5%(1/40) ,the difference was not statistically significant (P>0 .05) .Conclusion The combined treatment of Yunnan Baiyao Plaster in the patients with hemorrhagic stroke conld relieve pain and reduce the neurological deficit ,which might be related to the mechanism of anti-inflammation and analgesia .

Objective To address the medical problems, in treatment of avascular necrosis of the femoral head, on loosening and subsidence of the prosthesis of femoral head. A set of Mushroom Shaped Surface Prosthesis of Femoral Head was designed. Methods Based on biomechanical of the femoral head and the principle of transferring the force through femoral head while reducing stress-sheltered, nickel-titanium alloy and other new medical alloy materials were used to develop a prosthesis, which not only can be a weight-bearing device, but also promotes bone-growth. Results The prosthesis is novel, compact and little-volume, and the manipulation of surgery is convenient and secure. Conclusion The design of this prosthesis can provide new ways or ideas for designing the femoral head replacement prosthesis to solve problems on loosening and subsidence.

Objective To study the type variation of microglial activation in spinal dorsal horn of rats after sciatic nerve injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into the control and experimental groups, 24 rats in each group.The experimental group underwent ligation of sciatic nerve trunk to generate nerve injury in the rats.The pain behavior in the rats was measured at the 1th, 7th and 14th postoperative days, and the changes of microglial activation in the rat lumbar spinal cord dorsal horn was detected by immunofluorescence staining.qRT-PCR assay was used to validate the activation trends of M1 and M2 types of microglia cells.Results No significant changes were found in the microglial cells in the spinal cord dorsal horn of rats in the sham-operation group during 14 days after operation.In the sciatic nerve ligation group at 1 day after operation, no significant change was observed in the number of microglial cells, but the expression of marker of M1 microglia was significantly increased.At 7 and 14 days after operation, the number of microglial cells and the expression of M1 microglia marker in the spinal cord dorsal horn were increased significantly.Conclusions Microglia activation in the spinal dorsal horn starts at the first day after sciatic nerve injury, and lasts at least for two weeks after the operation.M1 microglia activation dominates during this period.

ObjectiveTo observe the effects of electroacupuncture on IL-1β in brain tissue of rats after total cerebral ischemia-reperfusion.MethodsThirty male Wistar rats were randomly divided into the sham operation group, model group and acupuncture group, in each group. In acupuncture group and model group, 4-vessel-occlusion method was used to make a rat model of cerebral ischemia reperfusion. Acupuncture group used electroacupuncture on points Quchi and Zusanli, 6 hours after reperfusion. The rats of each group were killed and ELISA was used for the measurement of IL-1β in brain tissues.ResultsIL-1β in brain tissue in model group increased significantly compared with sham operation group and acupuncture group(P<0-001). No significant differences were found between sham operation group and acupuncture group (P>0-05).ConclusionAcupuncture can lower IL-1β content of brain tissue of rats with brain ischemia/reperfusion injury.

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.

Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

ObjectiveTo investigate the efficacy of co-infusing cord blood (CB) as the third party cells on graft versus host disease (allo-GVHD) prophylaxis after unrelated or haploidentical donor allogeneic hematopoietic stem cell transplantation (allo-HSCT).MethodsFrom 2007 to 2011,41 patients receiving unrelated or haploidentical donor allo-HSCT were analyzed retrospectively.Twenty-five patients received one unit of HLA 4/6-6/6 matched CB one day before SCT as CB group,and median MNC dose was (1.64 ± 0.49) × 107/kg.Sixteen cases not receiving CB served as control group.All patients received antithymocyte globulin,cyclosporine,methotrexate,and mycophenolate mofetil as GVHD prophylaxis.The incidence and severity of aGVHD,and treatment-related mortality were compared between two groups.ResultsThe main clinical characteristics in both groups were comparable.The cumulative incidence of aGVHD in CB group and control group was 44.0％ versus 68.8％ respectively (x2 =2.403,P＞0.05).The cumulative incidence of grades Ⅲ to ⅣV aGVHD in CB group and control group was 16.0％ and 37.5％ respectively (x2 =2.445,P＞0.05).The 100-day treatment-related mortality in CB group and control group was 12.0％ and 12.5％ respectively (x2 =0.002,P＞0.05).ConclusionCord blood as the third party cells might reduce the incidence and severity of aGVHD in unrelated or haploidentical donor HSCT.The efficacy and the mechanism of this strategy need to be further explored by prospective randomized controlled trials.

Objective To evaluate the nucleated red blood cell (NRBC)count of Sysmex XN-9000 automatic hematology anal-ysis lines comparing with manual method,and verify the accuracy of the analyzer results.Methods 60 blood samples with more than 1% of NRBCs detected by XN-9000 were counted NRBCs by traditional manual microscopy in blood smears,and verified the analyzer results.Results According to the reliability analysis,the results of total 60 samples were all within the range of the reliability;correlation analysis showed that correlation coefficient (r)of group NRBC (%)1~10 and>10 were 0.972 1 and 0.996 2,respectively.There were no significant differences between them (P>0.05).Conclusion Compared with manual method,the results of NRBC count of XN-9000 were within the range of reliability,and showed good correla-tion.The analyzer test results of NRBC were accurate and reliable and could be applied to the detection of clinical samples.

Objective To investigate the expressions of CDX2 and β-catenin in acute lymphoblastic leukemia (ALL) and their correlation. Methods Real-time PCR was used to determine the expressions of CDX2 and β-catenin mRNA in 43 de novo ALL and 30 non-malignant patients (used as control). Results The positivity rate of CDX2 mRNA expression in ALL group was 93%, but CDX2 mRNA expression couldn′t be detected in the control group (P<0.01). The mRNA expression of β-catenin could be detected in patients in both two groups, butβ-catenin mRNA expression in the ALL group was significantly higher than in the control group (P < 0.01). And mRNA expressions of CDX2 andβ-catenin were significantly correlated with WBC counts and LDH level (P<0.01). When the ALL patients acquired complete remission (CR), the mRNA expressions of CDX2 and β-catenin were significantly decreased compared with their newly diagnosed status , while disease-relapsed the mRNA expressions of CDX2 andβ-catenin were increased again. There was significantly positive correlation between CDX2 and β-catenin mRNA expressions (r = 0.835, P = 0.000). Conclusion Up-regulation of CDX2 and activation of Wnt/β-catenin pathway coexist in the ALL patients and the mRNA expressions of CDX2 and β-catenin are positively correlated.