Objective To construct an expression vector harboring CYP2B1 suicide gene, and detect its expressions in tumor cell lines. Methods PCR amplification was performed using primers based on murine CYP2B1 gene sequence from gene bank and pc3/2B1 as template. PCR product was directly inserted an eukaryotic expression plasmid pcDNA3.0. The recombinants were analyzed and identified by restriction enzyme analysis, PCR and sequencing. Then the recombinant vector pcDNA3.0/CYP2B1 was transfected into three tumor cell lines by liposome-mediated method. The expressions of CYP2B1 gene in all the cell lines were detected by RT-PCR method. Results pCDNA3.0/CYP2B1 vector was successfully constructed, and could express CYP2B1 mRNA in the three tumor cell lines. Conclusion Eukaryotic expression vector pcDNA3.0/CYP2B1 containing CYP2B1 gene under the control of a CMV promoter is an novel effective expression vector for tumor gene therapy.

Objective To examine and evaluate the ability of spontaneous regeneration following various extent of incomplete injury in adult rats. Methods The quantities and reduplicated various extent incomplete injured model of adult rats optic nerve was established using different wounding time with constant wounding force produced by across action forceps at 2mm behind eyeball. GAP-43 (growth associated protein-43) and its mRNA expressions were detected with immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR). All data were analyzed by ANOVA. Results GAP-43 and its mRNA expression levels revealed that there was negative response at the distal area contrast to the strong positive response at the proximal in earlier period after injury until half mouth. One mouth later, the increased GAP-43 and its mRNA expression levels became more and more high and reached the climax at second month post injury. Then it decreased gradually. The result of RT-PCR showed there are significant difference among the various extent incomplete injured models and different time after injury. Conclusion Spontaneous regeneration of adult rat’s optic nerve can be detected and identified following incomplete injury, and the extent of regenerating ability is correlated with the extent of injury.

Objective To optimize the conditions for the extraction of volatile oil from Fructus Tsaoko and Radix Saposhnikoviae in Leifengguan granules,and for the preparation of their ?-CD inclusion compounds.Methods With the percentages of extracted volatile oil of Fructus tsaoko and Radix saposhnikoviae as the target marker,single-element experiments were conducted to determine the optimum conditions for extracting the volatile oil.The application rate of included volatile oil was used as the target marker to optimize the conditions for including the oils with ?-CD.Results The optimized extraction conditions were: with 8-fold water added,the sliced herbal medicine of Fructus tsaoko and Radix saposhnikoviae should be extracted for six hours without advanced soaking with water.The optimized inclusion conditions were: applying the saturated aqueous solution method,15-fold 85 % ethanol was used to dissolve the volatile oil,the proportion of the above solution and ?-CD being 1︰8;at 25 ℃,the volatile oil was included for 30 minutes with ultrasonic wave.Conclusion The established process can be used for the extraction and the inclusion of volatile oil from Fructus Tsaoko and Radix Saposhnikoviae in Leifengguan granules.

OBJECTIVE To evaluate the effects of arsenic trioxide (As2O3) on the cell cycle and microfilament cytoskeleton in human nasopharyngeal carcinoma cell line(CNE1),as well as possible mechanisms. METHODS The variation of cell cycle and microfilament cytoskeleton in CNE1 were observed using the flow cytometry (FCM),the laser scanning confocal microscopy(LSCM)and technology of fluorescence.RESULTS FCM showed that the proportion of G1 phase cells significantly increased in cells exposed to 2 and 4?mol/L As2O3(P

Objective To investigate the clinical efficacy of buttress plating for patients with posterior pilon fracture.Methods From April 2012 to January 2015,12 patients with posterior pilon fracture of the distal tibia were treated in our hospital.They were 7 men and 5 women,30 to 56 years of age (average,41.2 years).According to the CT classification by Haraguchi et al.,5 cases belonged to type I,3 to type Ⅱ and 4 to type Ⅲ.All the patients underwent open reduction and internal fixation with buttress plate via either a posterolateral approach or a dual posterolateral-posteromedial approach.All the patients were available for follow-up.The clinical outcomes were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and the visual analogue scale (VAS).The radiological evaluation was performed using the osteoarthritis-score (OA-score).Results The patients obtained an average follow-up of 21.2 months (range,from 12 to 30 months).Bone fractures united after an average of 15 weeks (range,from 13 to 19 weeks).The time for full weight walking averaged 16 weeks,ranging from 15 to 23 weeks.The ankle plantar flexion ranged from 36° to 42° (average,40.4°);the ankle dorsal extension ranged from 38° to 44° (average,42.6°).At the final follow-ups,the AOFAS scores ranged from 82 to 97 (average,88.2);the OA-score ranged from 0.6 to 0.8 (average,0.71);the VAS scores during rest,active motion and weight-bearing walking ranged from 0.5 to 0.8 (average,0.66),from 0.6 to 0.9 (average,0.82) and from 1.2 to 1.8 (average,1.41),respectively.No fracture malunion,implant loosening,pain or stiffness of the affected ankle was observed at the final follow-ups.Conclusion Buttress plating for posterior pilon fractures can lead to satisfactory clinical outcomes,because it ensures rigid fixation which in turn enables earlier postoperative mobilization.

Objective ToinvestigatetherelationshipbetweenmuscleRASoncogenehomolog (MRAS)genepolymorphismandyoungpatientswithischemicstroke.Methods Atotalof243young patients with ischemic stroke from Nanjing Stroke Registry Program from December 2009 to October 2012 were enrolled retrospectively. At the same time,512 age-and sex-matched healthy controls were selected. The polymorphisms of MRAS genes rs3755751 and rs9289559 loci were analyzed by the modified multiplex PCR-ligase detection reaction assay. The genotype of each locus and the allele frequencies were analyzed and compared. Results (1)The frequencies of AA,AG and GG genotypes for rs3755751 in the stroke group (n=243)were 7. 4%(n=18),37. 0%(n=90),and 55. 6%(n=135),respectively. There were no significant differences compared with those (6. 4%[n=18],36. 9%[n=189]and 56. 6%[n=190]) in the control group (n=512)(P>0. 05). The frequencies of AA,AG and GG genotypes for rs9289559 in stroke group were 7. 0 (n=17)%,42. 0%(n=102),and 51. 0%(n=124),respectively. There were no significant differences compared with 6. 1%,(n=31)37. 9%(n=194),and 56. 1%(n=287)in the control group (P>0. 05). (2)Further construction of an effect model (AA vs. AG+GG and GG vs. AG+AA),there was no significant difference between the stroke group and the control group (P >0. 05 ). Analyzing the effects of different genotypes on plasma lipid levels showed that the high-density lipoprotein cholesterol level of the GG genotype subgroup in the young ischemic stroke group was significantly higher than that rs3755751 of the AG +AA genotype subgroup (OR,6. 80,95%CI 2.18-21.27, P=0. 001 ). Conclusions MRAS gene polymorphism may have no significant correlation with the genetic susceptibility in young patients with ischemic stroke. Polymorphism of rs3755751 may be correlated with the level of serum high-density lipoprotein cholesterol.

AIM:To explore an ideal method to induce the differen-tiation of human umbilical cord mesenchy-mal stem cells (hUCMSCs) into neuron-like cells and to provide some evidence for the transplantation of hUCMSCs for spi-nal cord injury .METHODS:The hUCMSCs were isolated from human umbilical cord digested with collagenase Ⅱ.The hUCMSCs was verified by flow cytometry analysis .The passage 5 cells were randomly divided into 4 groups.The differentiation of hUCMSCs was induced by bFGF in group A , bFGF and BDNF in group B, or BHA, bFGF and BDNF in group C, while the cells in group D served as a control group cultured with DMEM-F12 and 10%FBS.Two weeks later , the expression of nestin , neurofilament protein H ( NEFH) and glial fibrillary acidic protein ( GFAP) was detected by real-time PCR and immunocytochemistry .The morphological changes of cells were observed under an atomic force microscope . RESULTS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by enzyme digestion .hUCMSCs expressed CD29, CD44 and CD105, but no CD34, CD45 or HLA-DR.After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuron-like cells.The appearance of the cells had great change .The induced hUC-MSCs developed round cell bodies with multiple neurite-like extensions observed under an atomic force microscope .The re-sult of real-time PCR showed that nestin was positive in A , B and C groups , and NEFH was positive in A and B groups , but GFAP was negative in 4 groups.The difference of nestin and NEFH expression among the induced groups was signifi -cant (P<0.05).CONCLUSION:Mesenchymal stem cells were isolated and cultured from human umbilical cord by en-zyme digestion in vitro, and all the hUCMACs presented stable biological properties .Moreover, hUCMSCs were induced to differentiate into neuron-like cells in vitro via bFGF combined with BDNF .

Objective To investigate the geographical distribution of plague host animals in residential areas and the association between plague and its distribution pattern in Yunnan Province.Methods A systematic investigation on fauna and community ecology of rodents was carried out in residential areas of 17 counties(cities) of Yunnan Province,southwestern China with different longitude,latitude and elevation from May 2007 to November 2012.The characteristics of the spatial distribution of flea communities along environmental gradients were analyzed using community structure indexes.Results A total of 390 small mammals were trapped in seventeen counties (cities),and the mammals were classified into 11 species and 7 genus in 4 families.Among all small host mammals,Rattus tanezumi,Rattus nitidus and Rattus norvegicus were dominant species of host animals in residential areas,accounting for 33.85％ (132/390),20.77％ (81/390) and 16.92％ (66/390),respectively.The horizontal distribution of rodents showed that Rattus tanezumi was the widest species in residential areas,which was found at the longitude 98°-105°,followed by Rattus norvegicus,Rattus yunnanensis and Mus musculus.The vertical and latitudinal distributions of rodents were similar in residential areas.Rattus tanezumi was also the widest distributed species,which was observed at the latitude 21°-＜ 28°N and at the altitude 500-＜ 3 500 m; the constituent of Rattus tanezumi showed similar trends of leaning peak curves,first gradually increasing and then decreasing with the increase of latitude and elevation; Rattus tanezumi gradually changed into Rattus nitidus and Apodemus chevrieri,et al.The richness spatial distribution patterns of host animals showed similar trends of leaning peak curves which gradually increased and then decreased with increasing of longitude and latitude; the highest richness was observed at the longitude 98°-＜ 101°E and at the latitude 23°-＜ 28°N.While,the richness spatial d istribution patterns of host animals were relatively poor in the low or the high altitude range.The highest richness was observed at the altitude between 1 000-＜ 1 500 m and 2 000-＜ 2 500 m.Conclusions All spatial patterns of indoor host animals have shown an aggregated spatial distribution in Yunnan Province.Moreover,important environmental factors such as longitude,latitude,and altitude play a substantial role on the distribution patterns of indoor host animals among various focus,therefore directly impact on the modes of prevalence and transmission of plague.

OBJECTIVE: Radial artery hemostasis devices of TR Band and RDP made in Japan has used in China and abroad. However,comparisons of application effects are rarely reported. The aim of this paper is to investigate the safety and effectiveness of radial artery hemostasis devices of TR Band and RDP after transradial intervention procedures. METHODS: A total of 300 cases with transradial route coronary arteriongraphy and percutaneous coronary intervention, were randomly divided into 3 groups, with 100 cases in each group. TR Band, RDP, and "8" character Bandaging were used respectively. The hemostasis effectiveness, hemostasis time, patients' degree of comfort, disorder of venous return,saturation of blood oxygen of thrums of compressed hands, as well as ischemia and necrosis status in local skin of access site were compared.RESULTS: Two kinds of devices were able to hemostasis effectively. The saturation of blood oxygen of cases in each group was within normal limits, which had no significantly difference (P> 0.05). Patients' degree of comfort was significantly different among three groups (P< 0.05), but there was no insignificantly different between TR Band and RDP groups (P> 0.05).Swelling in arms and ischemia and necrosis status of local skin at access site were notably different among three groups (P <0.05), however, ischemia and necrosis status of local skin at access site had no difference between TR Band and RDP groups (P>0.05).CONCLUSION: Application of radial artery hemostasis devices of TR Band and RDP-700(800) after transradial intervention procedures is safe and effective.

Objective To examine the frequency and mode of distal spread of low and middle rectal cancer in the mesorectum and rectal wall to determine the optimal distal clearance in situ. Methods Thirty-four specimens with low and middle rectal cancer were collected in the pathologic study between August 2004 and December 2005 in Cancer Center of Sun Yat-sen University,Twenty-eight specimens with low and middle rectal cancer were enrolled in the pathologic study between October 2006 and October 2007 in Shandong Provincial Hospital of Shandong University.Logistic regression wag used to analyze clinicopathoiogic factors related to distal spread of low and middle rectal cancer in the mesorectum and rectal wall. Results Two types of disial spread of the tumor were identified in rectal wall:submucosa invasion and muscularis propda invasion.Distal spread in rectal wall was observed in 16%(10/62)of the patients.The length of distal spread in rectal wall was found from 0.5 cm to 1.0 cm.Four types of distal spread of the tumor were identified in mesorectum:lymph node invasion,blood and lymphatic vessel invasion,perineural invasion,isolated neoplastic microfoci.Distal spread in mesorectum was observed in 19%(12/62)of the patients.The length of distal spread in mesorectum was found from 0.5 cm to 4.0 cm.Univariate analysis showed that serum CEA,lymph node invasion.circumferential margin involvemenl and Dukes stage were correlated with distal spread of low and middle rectal cancer in the mesorectum and rectal wall.Dukes stage was shown to be independent impact factor by multivariate analysis(Wald=8.386,P=0.004).Conclusion Dukes stage is an independent impaet factor for distal spread of low and middle rectal cancer in the mesorectum and rectal wall.Resection of 1.5 cm for distal rectal wall mandatory for a curative resection,provided that the clearance for distal mesorectum is no less than 5.0 cm.