Objective To investigate the genetic relationship of the Dengue virus strains isolated in Guangdong province in 2006 and 2007, and to find the sources of these virus. Methods Serum samples of the dengue fever patients from 5 cities of Guangdong province in 2006 and 4 cities in 2007 were collected. Three pairs of primers that specific for amplifying the 3 overlap fragments of E gene of Dengue virus type Ⅰ were designed. RNAs were extracted from C6/36 cells treated with patients' serum. E genes were amplified by RT-PCR, purified and then sequenced directly. To obtain the E gene complete sequences, the raw sequences were assembled and edited. Obtained E genes were compared with E genes of other Dengue virus type Ⅰ published in the GenBank, analyzed by MEGA version 4.1 software. Results In 2006, virus circulating in Guaogzhou2006(EF508203) was closest to Vietnam2006(EU482539) with 99.3% nucleotides homology, Chaozhou2006 (EF508206) and Shantou2006 (EF508207) strains were closest to Japan2004 (AB178040) and Singapore2006 (EU081280) with 99.5% nucleotides homology, while Yangjiang2006 (EF508205) and Yangjiang2001 (EF508200) were closest to each other and both with 99.5% nucleotides homology to Thailand2001 (AY732482). All 4 Dengue virus strains circulated in 2007 were closest to Singapore2005(EU081276) with 99.7% nueleotides homology. Conclusion The Dengue viruses prevalent in 2006 were from different sources while those in 2007 came from the same origin. The data also showed that there was an endemic area of Dengue virus in Guangdong province.

Objective:To study the effects of tamoxifen on proliferation and ER expression of human hepatocellular carcinoma cells.Methods:HepG2 cells were treated with tamoxifen at different concentration and different action time.MTT was used to determine the suppression rate of human hepatocellular carcinoma cell.The effects of tamoxifen on human hepatocellular carcinoma cell ER performance were observed by immunohistochemistry.Results:Tamoxifen inhibited the proliferation of human hepatocellular carcinoma cells and suppressed human hepatocellular carcinoma cell ER performence.Conclusions:Tamoxifen may suppress human hepatocellular carcinoma cell proliferation ER performance.

Objective: To detect the expression of sparc/osteonectin, cwcv and kazal-like domains proteoglycan 1 (SPOCK 1) gene in bladder cancer cells, and further to investigate the effects of down-regulation of SPOCK 1 gene expression on the proliferation, migration and invasion of bladder cancer cells. Methods: The expression level of SPOCK 1 gene in human ureteral epithelial immortalized cell line SV-HUC-1 and bladder cancer cell lines 5637 and T24 was detected by real-time fluorescent quantitative PCR (RFQ-PCR) and Western blotting, respectively. The siRNA fragments targeting SPCOK 1 gene were constructed and transfected into bladder cancer 5637 cells by liposome (as the experimental group); at the same time, the 5637 cells were transfected without anything as the blank control group and transfected with scramble siRNA as the negative control group. Then the expressions of SPOCK1 mRNA and protein in each group were detected by RFQ-PCR and Western blotting, respectively. The changes of proliferation, migration and invasion abilities of bladder cancer 5637 cells after SPOCK1 expression down-regulation were detected by CCK-8 method, Wound healing assay and Transwell chamber assay, respectively. Results: As compared with the ureteral epithelial immortalized cells, the expression levels of SPOCK1 mRNA and protein were higher in bladder cancer cells, especially in 5637 cells (both P < 0.05). After transfection with SPOCK1 siRNA, the expression of SPOCK 1 gene was downregulated (P < 0.05), the proliferation, migration and invasion of bladder cancer 5637 cells were significantly inhibited (all P < 0.05). Conclusion: SPOCK 1 gene is overexpressed in bladder cancer cells. The down-regulation of SPOCK 1 gene expression can reduce the proliferation, migration and invasion of bladder cancer 5637 cells.

In this study, the bioactivity of a novel BMP2-derived oligopeptide P24 was investigated by using the model of rabbit femoral defect after loaded in the biodegradable poly (lactic acid / glycolic acid / asparagic acid-co-polyethylene glycol) (PLGA-[ASP-PEG]). A 1.5-cm unilateral segmental bone defect was created in the left femoral diaphysis in each of the 30 new zealand white rabbits. The defects of 18 legs filled with BMP2-derived peptide P24 combined with PLGA-[ASP-PEG] scaffold serves as the experimental group, and the defects in the rest 12 rabbits filled with (PLGA-[ASP-PEG]) without P24 as control group. The bone-repairing capability in the target region of the two group was grossly, radiologically, histopathologically and biomechanically evaluated 4, 8 and 12 weeks after the operation. Our results showed that in each group, primary healing of incision was achieved in the two groups. Radiographically, in experimental group, defects were filled with induced callus within 8 weeks, and a cortical bone-like structure was observed in some animals at the 12th week. According to the standardized stage of bone defect repair, 9 (64.28%) achieved grade-4 healing. In contrast, little bone formation was seen in the defects even 12 weeks after the operation, and 5 (62.50%) had grade 0 healing in this group. Histologically, tissue engineering material was mostly absorbed and cartilage was found around implants in the experimental group at the 4th week; 8 weeks after operation, the engineering material was completely absorbed, and formation of woven bone was observed and typical trabecular bone structure could be seen. In control group, 8 weeks after operation, the defect was filled with fibrous tissues, and no bone-like structure was observed. Statistical analysis showed very significant difference in biomechanical indicators between the two groups (P<0.05). It is concluded that new oligopeptide P24 can induce excellent bone regeneration and promote bone repair.

Objective To study the factors influencing epidemiological characteristics and virulence of Enterovirus 71 (EV71) in Guangdong province from 2008 to 2010. Methods RNA was extracted from collected samples or cultured virus , then reversing transcription into cDNA. We amplified full-length EV71-VP1 using poly-merase chain reaction technology , then conducted sequence alignment and established phylogenetic tree with MEGA software (version 5.0) to confirm the genotype of EV71. The association between severity of clinical symp-toms and sex, age, viral genotype and VP1 variation was also analyzed using Logistic regression. Results The genotype of the predominant epidemical strain was C4a in Guangdong from 2008 to 2010. However , this subtype had already differentiated into 4 subgroups (C4a1- C4a4). There was no correlation between clinical syndrome and sex or viral genotype; the severity of symptoms was negatively correlated with age: before 4 years old, varia-tion A289T can easily lead to severe cases, increasing the risk of infection (P<0.05, OR = 2.360, 95%CI:1.163~ 4.659). Conclusion The main epidemical EV71 strain is C4a1 in Guangdong province. The emerging differen-tiation and simultaneous prevalence should merit attention to strengthen relevant surveillance; and the protection of the susceptible population should be reinforced during EV71 prevalence.

Objective To explore the preparation and quality control of As2 O3 nanoparticle .Methods PEG‐PLA was used as the vector material to prepare As2 O3 nanoparticle with ultrasonic emulsification method ,and the VEGFR‐2 was coupled to obtain VEGFR‐2/As2 O3‐PEG‐PLA nanoparticle .The particle size distribution ,Zata potential ,loading efficiency (LE) ,encapsulation effi‐ciency(EE) ,drug release in vitro and stability was determined ,and morphological characteristics was observed by transmission elec‐tron microscope(TEM) .Tweety‐four hepatocellular carcinoma nude mices were randomly divided into VEGFR‐2/As2O3‐PEG‐PLA nanoparticles group and As2 O3‐PEG‐PLA nanoparticles group ,by tail vein injection of nanoparticles .High performance liquid chro‐matography was used to determine content of As2 O3 .After 21 d ,six nude mices in each group were killed ,and the immunohisto‐chemistry and western blot method was used to detect the expression of VEGFR‐2 .Results The particle size of VEGFR‐2/As2 O3‐PEG‐PLA was determined to be (141 .9 ± 13 .2)nm ,Zata potential was (10 .2 ± 1 .1)mV .It was found to spherical or oval shape , with uniform size and dispersibility under TEM .LE and EE was (5 .51 ± 1 .83)% and (62 .12 ± 5 .98)% ,respectively .Drug release in vitro showed that VEGFR‐2/As2 O3‐PEG‐PLA exhibited controlled release effect ,with half of the release time as 10 h .Besides , VEGFR‐2/As2 O3‐PEG‐PLA showed a good stability in 3 days .Compared with As2 O3‐PEG‐PLA nanoparticles group ,the concen‐tration of As2 O3 in tumor and liver tissue was high ,the concentration of As2 O3 in blood ,heart ,kidney tissue was low ,the expression of VEGFR‐2 in tumor tissue was low in VEGFR‐2/As2O3‐PEG‐PLA nanoparticles group(P< 0 .05) .Conclusion The prepared As2 O3 nanoparticle using PEG‐PLA as vector and VEGFR‐2 as target showed uniform size ,high EE and LE ,good stability .And it preliminarily proved that VEGFR‐2 could be targeted in nude mice .

. Conclusions Two multiplex RT-PCR assays show high consistency with common RT-PCR. The multiplex RT-PCR assays were initially established.

Objective To establish a method for the isolation of Zika virus and to gather experi-ences for viral isolation. Methods Suckling mice at age 1-3 days were inoculated with serum samples posi-tive for Zika virus through intracranial injection. All mice were sacrificed 6 days after the injection. Viral nu-cleic acids were extracted from brain, heart, liver, spleen, lung, kidney, muscle, skin and intestine tissue samples and analyzed by real-time RT-PCR. The supernatants of brain tissues positive for Zika virus were used for subculturing. Nested PCR was performed to amplify the NS5 gene of the isolated virus. The se-quences of NS5 gene were analyzed by using MEGA6. 0 software. Results All of the tissue samples were positive for Zika virus. Higher viral loads were detected in heart and brain tissue samples with cycle thresh-old (Ct) values of 24. 4 and 25. 3, respectively. The second generation of Zika virus was identified in suck-ling mice brain tissues 2 days after infection by using real-time RT-PCR. The amplified product of nested PCR was 972 bp in length. Sequencing analysis showed that the isolated Zika virus ( GDZ16002 strain) be-longed to the Asian lineage. Conclusion A strain of Zika virus was successfully isolated in China by using intracranial injection via a suckling mouse model. The isolated Zika virus belonged to the Asian lineage.

Objective To investigate the expression and clinical significance of β2 microglobin (β2-MG)protien and vascular endothelial growth factor (VEGF) protien in diffuse large B-cell lymphoma (DLBCL).Methods The expressions of VEGF protien and β2-MG protien were evaluated in 49 DLBCL patients which started the initial treatment by luminex suspension array.Results Among 49 DLBCL patients,expression of β2-MG protein was high in 37 cases and the expression of VEGF protein was high in 23 cases.The expression of VEGF protien and β2-MG protien were not related with gender,age,B symptoms,clinical stage and lactic acid (LDH).There was positive correlation between the high expression of β2-MG protien and chemotherapy (P =0.037).There was relevant trend between the higher expression of VEGF protien (P =0.067).Conclusion The expressions of VEGF protien and β2-MG protien are detected in DLBCL,both proteins may be the potencial markers of DLBCL and therapeutic targets for DLBCL.