Objective To establish a nude mouse model of low rectal cancer with lymph node metastasis, to detect the expression of E-calcium (E-cad) in the serum and transplanted tumor tissue of the mice, and to analyze the role of E-cad in lymph node metastasis of rectal cancer.Methods Human colorectal adenocarcinoma cells (SW-480) were inoculated into the rectal submucosa of nude mice to establish a model of low rectal cancer with lymph node metastasis.120 nude mice were randomly divided into three groups (40 mice in each group).Group A (rectal cancer model group) was injected with dimethylhydrazine and implanted with SW480 cells.Group B (dimethylhydrazine control group) was injected with dimethylhydrazine alone.The group C (control group) received no treatment.Serum E-cad was detected by ELISA.The mRNA and protein expression of E-cad in the tumor tissue was detected by RT-PCR and western blot, respectively.Results The serum level of E-cad in group A was 19.48±1.25 mg/L, significantly higher than those in groups B (2.36±0.18 mg/L) and C (2.15±0.12 mg/L) (t =8.28, 9.01, P < 0.05).The serum levels of E-cad were significantly higher in the nude mice with lateral lymph node metastasis than those without lymph node metastasis (t =10.28, P < 0.05).The protein expression of E-cad in the group A was lower than that in the groups B and C (t =9.81, 7.69, P < 0.05).In the group A, the protein expression of E-cad in the nude mice with lateral lymph node metastasis was significantly lower than that without lateral lymph node metastasis (t =9.36, P < 0.05).The mRNA expression of E-cad in the group A was significantly lower than that in the groups B and C (P < 0.05), and the mRNA expression of E-cad in the nude mice with lateral lymph node metastasis was significantly lower than that in the mice without lateral lymph node metastasis (t =7.85, P < 0.05).Conclusions The serum level of E-cad may be closely associated with the lymphatic metastasis of rectal cancer, and the mRNA and protein expressions of E-cad in colorectal cancer tissue were weak or absent, leading to a decrease of adhesion ability between cancer cells, and promote the invasion and metastasis of cancer cells.

Objective To evaluate the dose distribution of target volume and normal tissues with different treatment planning such as three dimensional conformal radiotherapy(3DCRT) ,simplified intensity modulated radiotherapy(sIMRT) ,and intensity modulated radiotherapy (IMRT) for patients with radically resected rectal cancer. Methods Ten male patients with stage Ⅱ and Ⅲ rectal cancer after radical resection (Dixon surgery) were enrolled in this study. 3-field or 5-field 3DCRT,slMRT and 5-field or 7-field IMRT plans were performed for each patient. The dose distributions of target volume and normal tissues,conformal index(CI) and heterogeneous index(HI) were analyzed using the dose-volume histogram(Dvit). The prescription dose was 50 Gy in 25 fractions. Results The CI for PTV of IMRT and sIMRT was superior to 3DCRT. Conversely,the HI for PTV of 3DCRT was superior to sIMRT and IMRT. sIMRT and IMRT can protect the organs at risk better than 3DCRT. The mean of total MU for 3DCRT3f,3DCRT5f, sIMRT, IMRT5f and IMRTT7f was 482±13,504±11,455±42,841±36 and 884±46, respectively. Conclusions Comparing with 3DCRT plans and IMRT plans, sIMRT plan was the optimal plan for clinical practice. All of the three radiotherapy techniques can protect the rectal stump and anal canal well with the prescription dose of 50 Gy.

It is generally believed that liposomes modified with polyethylene glycol (PEG) have no or lower immunogenicity. However, based on many recent literatures, when the PEGylated liposomes were repeatedly applied to the same animal, the immune responses occurred. The first injection of PEGylated liposomes resulted in a reduction in the circulation time and an increase in hepatic and splenic accumulation of the second dose of PEGylated liposomes in a time-interval, which was called "accelerated blood clearance (ABC)" phenomenon. Such immunogenicity of PEGylated liposomes presents a barrier in the research of liposomal formulations and their use in the clinics. This review focused on the definition, the method of verification, the development of the reason for ABC phenomenon, influencing factors of ABC phenomenon, and discussed if other PEGylated nanocarriers also induce ABC phenomenon.

Objective To study the clinical, radiologic and pathologic characteristics and diagnostic methods of telangiectatic osteosarcoma for further improving the diagnostic ability.Methods The data of 10 patients with histologically proved telangiectatic osteosarcomas were retrospectively reviewed, and the clinical, radiologic and pathologic characteristics were further analysed in combination with the literature.All 10 patients were examined with X-ray and MRI, and 2 patients with CT.Results Telangiectatie osteosarcomas originated from inferior femur in 5 patients, femur neck in 1 patients, superior humerus in 2 patients and superior segment of tibia in 2 patients.The lesions showed osteolytic bone destruction on X-ray films (n = 10) and CT images ( n = 2), with mild bone expansion in 4 patients.The majority of the edge of the destroyed bone areas was unclear but without sclerotic rim.There were Codman's triangle and soft tissue mass in each patient but no obvious neoplastic bone forming.On MRI, all the lesions were mostly or completely constituted by the multiple cysts with periostnal reaction, and several scatteredly smaller liquid-liquid levels were found within cystic cavity in 7 patients.In all 10 cases, there were pathologic hemocoele similar to aneurysmal bone cysts (ABC), but malignant tumor cells and some neoplastic bones were found in cystic walls or septations.Only a small number of neoplastic bone tissue were seen by microscopy in 6 patients.Conclusions The radio-pathologic characteristics of telangiectatic osteosarcoma include the similar imaging findings of ABC, the common growth patterns of malignant tumors,and the pathologic hemocoele, malignant tumor cells within cystic wails or septations.The comprehensive analysis of clinical, radiologic and pathological data may help clinicans to make a correct diagnosis for telangiectatic nsteosareoma.

Objective:To explore the effect of calcium release-activated calcium channel modulator 1 (ORAI1) on the migration and invasion of colon cancer cell line SW480 and its mechanism. Methods:The SW480 cells were infected with ORAI interference lentivirus. The expression of ORAI1 mRNA and protein was confirmed by quantitative real-time polymerase chain reaction and Western blot. Transwell chamber, adhesion, angiogenesis, and vasculogenic mimicry experiments were conducted to detect the ability of cell invasion, migration, and angiogenesis and the intercellular adhesion of homogeneous and heterogeneous cells among each group. Confocal microscopy was employed to detect the difference of store-operated Ca2+entry (SOCE) in each group. Western blott was used to detect the expression of ERK1/2, p-ERK1/2, MMP-2, VEGF, and E-cadherin protein. Results:After the infection of SW480 with the ORAI1 interference lentivirus for 72 h, significant fluorescence expression was observed. Compared with the empty vector group and control group, the expression of ORAI1 was lower in the interference group (P<0.01). Invasion and migration ability decreased (P<0.01); the intercellular adhesion ability of homogeneous cells increased (P<0.05); the intercellular adhesion ability of heterogeneous cells decreased (P<0.05);the angiogenesi and vasculogenic mimicry were enhanced (P<0.01);the internal flow peak of SOCE was low (P<0.05); the expression of p-ERK1/2, MMP-2, and VEGF proteins decreased (P<0.01); and the expression of E-cadherin protein increased (P<0.01). Conclusion:ORAI1 may promote the migration and invasion of SW480. This mechanism may be associated with the increase of SOCE.

Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P <0.05 ) , homogeneous cells intercellular adhesion increased and heterogeneous cells intercellular was opposite ( P <0.05 ) , meanwhile , the expression of E-cadherin was declined significantly ( P<0.05 ) ,the expression of N-cadherin , vimentin raised significantly ( P<0.05 ) .Conclusions Silencing Gli2 gene can reverse the EMT of the hepatocellular carcinoma cell line SMMC-7721 and inhibit cell invasion ,its mecha-
nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.

Objective To understand the genetic characteristics of Enterovirus 71 ( EVT1 ) circu-lating strains of Guangdong province in 2008. Methods We isolated an EV71 strain from the fatal case of the hand, foot and mouth disease during an epidemic of Guangdong in 2008. Its complete genome was se-quenced and analyzed comparatively. Results The results showed that the full length of EV71 GDFS-3 ge-nome( not including poly A tail ) is 7405 bp. No insertion or deletion is detected in the coding region. There are several insertions and deletions in 5'and 3'UTR. Phylogenetic analysis of GDFS-3 and reference strains showed GDFS-3 strain shares the highest nueleotide homology with TW984 strain(96.0% ) but low homology with SIN5865, MS and BrCr( about 81.0% ). GDFS-3 strain also shares the highest amino acid homology with TW984 strain(99.0% ). It clustered with reference strains of CA subgenotype in the phylogenetie tree. The nucleotide identity with CA reference strains is 91.0% -95.0%. Conclusion The phylogenetic analysis based on the entire genome demonstrates that GDFS-3 strain has the nearest genetic relationship with TW984 strains ( isolated in 2004). GDFS-3 may belong to the same subgenogroup ( CA ) with Taiwan predominant strains. Otherwise,Some mutations in 5'UTR of EV71 may play the very important role in heightened viru-lence.

Objective To study the molecular characters of porA and porB genes which encode outer membrane proteins (OMP), and predominated clonal complex of Neisseria meningitidis isolates from Guangdong province. Methods Eighteen Neisseria meningitidis isolates from Guangdong province during year 1967 to 2007 were recovered and reconfirmed by API NH biochemical system, and serogrouped by antiserum. The characters of porA and porB gene were analyzed by DNA sequencing. The allele profiles and the sequence types (ST) were determined by multilocus sequence typing (MLST). Based on their allelic profiles, the evolution relationship was analyzed by PHYLIP software. The predominant clonal complex was determined through comparing with the information of reference strains from the PubMLST database. Results For porA gene, type 20 was more frequently in the variable region (VR) 1 and type 9 in VR2 before year 2004. However, for porB gene, type 4 was more frequently in VR Ⅰ, type7 in VR Ⅳ, type 11 in VR Ⅴ,and type 10 in VR Ⅵ, respectively. The multi-types character was presented in VR Ⅴ and VR Ⅵ after2004. VR Ⅶ and VR Ⅷ can not be found among all the isolates except for one W135 isolate in 2007. Among the seven housekeeping genes, the polymorphism of abcZ was the lowest one with 4 allele numbers, while pgm was the highest one with 13 allele numbers. The predominant clonal complex was ST-5 before 2004. The ST-4821 clone complex appeared in 2004 and caused cases every year since then. More important, highly invasive ST-11 clonal complex firstly appeared in Guangdong in 2007. Conclusion The molecular characteristic of OMP genes presents polymorphism for the Neisseria meningitidis isolates from patients in Guangdong province during 1967 to 2007. ST-5 is the predominant clonal complex before 2004 and the highly invasive clonal complex is circulating in recent 3 years. It suggests that the surveillance based on laboratory should be further enhanced.

BACKGROUND: Strontium-doped calcium polyphosphate (SCPP), as a new repairing material, has been demonstrated to have favorable biocompatibility and biodegradability and some effects on promoting self-angiogenesis. However, the mechanism remains still unknown. OBJECTIVE: Endothelial cells were cultured with SCPP scaffolds in vitro, as well as the cell proliferation and angiogenic factor matrix metalloproteinase-2 (MMP-2) secretion were observed. DESIGN,TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from September 2008 to April 2009. MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared.METHODS: ① Materials were plated on 24-well culture plate,and endothelial cell suspension (300 μL) were seeded on 24-well culture plate at the concentration of 3×10~7/L and cultured with 200 uL RPMI1640 culture media. Endothelial cell proliferation was observed using MTT method at days 1,3,5, and 7 after culture. ② CPP and 8% SCPP were plated on 24-well culture plate, and endothelial cell suspension (300 uL) was then incubated in 24-well culture plate at the concentration of 1x10~8/L and cultured with 600 uL RPMI1640 culture media. The morphology of endothelial cells was observed by scanning electron microscopy (SEM) at day 5 after culture.③ Endothelial cells were co-cultured with SCPP of various Sr~(2+) contents for 5 days. After confluence, cells were centrifuged to obtain supernatant. Angiogenic factor MMP-2 secretion was evaluated by ELISA assay.MAIN OUTCOME MEASURES: The proliferation and morphology of endothelial cells on SCPP and CPP were observed. The amount of endothelial cells-derived MMP-2 protein secretion was detected. RESULTS: MTT method demonstrated that the proliferation of endothelial cells on the 8% SCPP scaffold showed a higher level compared to CPP, and other SCPP groups. Scanning electron microscope results suggested that endothelial cells on 8% SCPP had a better growth status and biological activity. ELISA results indicated that angiogenic factor MMP-2 expression on the SCPP was promoted compared with that of CPP, and 8% SCPP showed the highest expression (P < 0.05). CONCLUSION: SCPP has good compatibility with endothelial cells,promoting angiogenesis and enhancing the angiogenic factor MMP-2 expression.

Objective To explore the effectiveness of our self-designed 3D printing relay template in the treatment of malunion of irregular intra-articular ankle fractures.Methods From October 2013 to October 2015,our self-designed 3D printing relay template was used in 14 patients with severe malunion of intra-articular ankle fracture to assist their intra-articular osteotomy.They were 10 males and 4 females,with an average age of 40.5 years (from 34 to 59 years).Thin slice CT scan was preformed preoperatively for all the ankles involved.The computer fictitious tibial and fibular models for location and osteotomy navigation templates were acquired from the data of CT scan after processing,design and reconstruction by software.After the navigation templates which fit the size of solid ankle were printed by a 3D printer,they were sterilized and reserved.The guide plates were used in turn during operation.The irregular malunited bone blocks in the posterior malleolus were completely resected along the osteotomy template.After fracture reduction,the bone blocks were stabilized with internal fixation.Fracture union time was recorded.Scores of visual analogue scale (VAS),American Orthopaedic Foot and Ankle Society (AOFAS) and the short form-36 (SF-36),ankle ROM and complications were also recorded at the final follow-ups.Results The 14 patients obtained an average follow-up of 28.6 months (from 14 to 35 months).Bony union was achieved after an average of 3.7months (from 3 to 6 months).At the final follow-ups,on average,VAS score (2.2 ± O.8),AOFAS ankle and hindfoot score (81.1 ± 6.8),SF-36 score (82.5 ± 5.2),dorsal extension (27.5 ° ± 2.5°),and plantar flexion (24.2° ±6.3°) were significantly improved compared with the preoperative values (6.3 ± 1.8,43.1 ± 12.0,37.6 ± 9.9,14.1° ± 3.8° and 14.4° ± 3.9°,respectively) (P ＜ 0.05).No implant failure,soft tissue complications or post-traumatic arthritis occurred during the follow-up.Conclusion Since our self-designed 3D printing relay templates for osteotomy can accurately position the fracture lines of the distal tibia via limited incision and invasion,they help a lot in precise osteotomy and anatomical reduction of malunited intra-articular ankle fracture,preventing incidence of traumatic arthritis.