OBJECTlVE To study the anti-HBV activity of prepared recombinant human serum aIbu-min-interferon α-2b fusion protein(HSA-IFNα-2b) in vitro. METHODS HepG2 ceIIs were infected with recombinant adenovirus with green fIuorescence protein and 1.6-foId HBV DNA(AdGFP-HBV). The ex-pression of HBV antigens,HBsAg and HBeAg in cuIture medium was detected by ELISA assay. The tox-icity of HSA-IFNα-2b on HepG2 ceIIs was evaIuated by mTT assay.The reIative expression of HBV RNA in ceIIs and the absoIute quantity of HBV DNA in cuIture supernatant were determined by quantitative PCR assay. The activity of HBV enhancer Ⅰ was detected by DuaI-Reporter gene assay. RESULTS HBV couId repIicate and express in HepG2 ceIIs after infection with AdGFP-HBV. The expression of HBsAg and HBeAg in cuIture serum of HepG2 ceIIs infected with AdGFP-HBV decreased by 51.32%(P﹤0.01)and 50.26%(P﹤0.01),respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The same concentration of HSA-IFNα-2b didn't inhibit the proIiferation of HepG2 ceIIs,but inhibited HBsAg in a concentration-dependent manner. The regression formuIa between HBsAg inhibitory rate(Y)and con-centration of HSA-IFNα-2b(X)was Y=21.11 IgX+11.91(r 2 = 0.954),IC50 = 63.76 kU·L-1 . HBV RNA in ceIIs and HBV DNA in the cuIture serum decreased by 52.83%(P﹤0.01)and 53.07%(P﹤0.01), respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The activity of enhancer Ⅰ decreased by 40.04%(P﹤0.01)when HSA-IFNα-2b 500 kU·L-1 was added. CONCLUSlON The ceII modeI of HBV repIication for evaIuating anti-HBV agents is successfuIIy estabIished. HSA-IFNα-2b exhibits noticeabIe anti-HBV effect invitro.

Objective To compare and analyze the clinical characteristics of acute central cervical spinal cord injury with only upper extremity involvement and with both upper and lower extremity involvement.Methods A retrospective case control study was made on clinical data of 76 patients with acute central cervical spinal cord injury hospitalized from January 2010 to December 2013.Nerve injury involved was only upper extremity in 39 patients (upper extremity group),but both upper and lower extremities in 37 patients (upper-and lower-extremity group).In upper extremity group,there were 35 males and four females,age was 21-80 years [(52.5 ± 13.4) years],injury resulted from traffic accidents in 24 patients,ground-level falls in eight,high-level falls in six and heavy-object hit in one,and level of injury was C3/4 in 16 patients,C4/5 in 14 and C5/6 in nine.In upper-and lower-extremity group,there were 30 males and seven females,age was 36-78 years [(59.6 ± 9.7) years],injury resulted from traffic accidents in 16 patients,ground-level falls in 11,high-level falls in seven and heavyobject hit in three,and level of injury was C3/4in nine patients,C4/sin 18 and C5/6in 10.Sagittal diameter of the cervical spinal canal,maximal canal compromise,maximal spinal cord compression,degenerating factors of the cervical spine and treatment protocols were determined.Upper extremity function was assessed with the American spinal injury association (ASIA) score.Results There were significant differences between upper extremity group and upper-and lower-extremity group in sagittal diameter of the cervical spinal canal [(7.5 ± 1.5)mm ∶ (6.8 ± 1.2)mm],maximal canal compromise [(28.9 ±9.6)％ ∶ (34.9 ± 10.6)％],ASIA score at admission[(31.6 ± 11.8)points ∶ (22.7± 11.3)points)] and ASIA score at last follow-up [(46.2 ± 4.2) points ∶ (40.2 ± 4.0) points] (P ＜ 0.05),while the maximal spinal cord compression in upper extremity group [(15.7 ± 11.9)％] had no significant difference from that in upper-and lowerextremity group [(17.0 ± 10.6) ％] (P ＞ 0.05).Lower prevalence of posterior osteophyte of the vertebral body was noted in upper extremity group than upper-and lower-extremity group (15％ ∶ 51％) (P ＜0.01).Twenty patients (49％) in upper extremity group were surgically treated,while 31 patients (84％) in upperand lower-extremity group (P ＜ 0.05).Conclusions Compared to acute central cervical spinal cord injury with both upper and lower extremity involvement,the injury with only upper extremity involvement is much common in younger patients and is characterized by lowered frequency of osteophyte,large buffer space,mild nerve damage,preferred non-operation treatment and good prognosis.

OBJECTIVETo construct a dual-reporter gene system that will be applicable for use as a tool to screen and evaluate therapeutic drug compounds that inhibit transcription of the gene encoding collagen I, chain at1 (COL1A1).

METHODSThe full-length eDNA of transforming growth factor beta1 (TGFbeta1) was cloned by RT-PCR and inserted into two vectors, pcDNA3.1 and pJW4303, for construction of two eukaryotic expression vectors, pcDNA3.1-TGFbeta1 and pJW4303-TGFbeta1.Next, the promoter region of COL1A1, cloned by PCR using human genome DNA as template, was inserted into the vector pGL4.29 to construct the reporter gene vector, pGL4.29-COL1A1 promoter.All three recombinant vectors were verified by restriction enzyme digestion and DNA sequencing.Either the pcDNA3.1-TGFbeta1 or pJW4303-TGFbeta1 vector along with the pGL4.29-COL1A1 promoter vector or a pRL-null, control reporter, vector were co-transfected into the LX-2 human hepatic stellate cells to establish the transcription-activated dualreporter gene system.This system was used as a cell model for screening anti-liver fibrosis compounds that inhibit the transcription of COL1A1.Dexamethasone, a model drug that is known to inhibit the expression of COL1A1, was used as a control to validate the dual-reporter gene system.

RESULTSThe two TGFbeta1-expressing vectors and the reporter gene vector containing the promoter region of COL1A1 were successfully constructed.The results of a dual-reporter gene assay showed that TGFbeta1 co-expression increased the activity of the COL1A1 promoter by above 200-fold (t =21.78, P =0.0001), whereas in the absence of TGF31 co-expression the activity was below 2-fold (t =3.396, P =0.0274).The transcriptionactivated dual-reporter gene system was successfully established.The model drug, dexamethasone, effectively inhibited the activity of the COL 1A1 promoter in dose-dependent manner; the activity decreased 29.6% with 10 mumol/L dexamethasone (t =4.140, P =0.0144) and 53.9% with 100 mumol/L (t =6.193, P =0.0035).

CONCLUSIONThe dual-luciferase reporter system of TGFbeta1 and COL1A1 co-expression developed here can be used as a cell model to screen and evaluate anti-liver fibrosis compounds that inhibit activity of the COL1A1.

Base Sequence ; Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; Genes, Reporter ; Genetic Vectors ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Transfection ; Transforming Growth Factor beta1

Objective: To understand the current situation and associated factors of myopia in junior middle school students, and to provide scientific basis for prevention and control of myopia in junior middle school students. Methods: A total of 5 393 junior middle school students were selected from middle schools in Guangdong, Guangxi, Guizhou, Liaoning, Shandong, Shanxi provinces. The visual acuity of middle school students was examined, and the data of general population, economy, sociology and natural environment were obtained through statistical yearbook of each province. The influencing factors of myopia of middle school students were analyzed by univariate and multivariate Logistic regression. Results: The results of single factor analysis showed that the myopia rate of junior high school students was different by gender, grades, parents average wage, sunshine duration, temperature, altitude, longitude and latitude(χ2=47.76，59.05，10.79，106.19，53.56，85.02，76.23，107.07，P<0.05). The results of multi factor analysis showed that gender, grade, average wage, temperature and latitude was positively associated with myopia vision; sunshine duration and longitude were negatively associated with the risk for myopia(OR=1.54，1.34，1.62，7.58，27.10，0.42，0.39，P<0.05). Conclusion The myopia of junior high school students is affected by a variety of factors, economic and social factors and natural environmental factors have an impact on the screening of sexual myopia in junior high school students. Economic and social factors and natural environmental factors should be taken into account in the formulation of myopia prevention and control measures.

Objective: To understand the current situation and associated factors of myopia among primary school students, and to provide scientific basis for prevention and control of myopia among primary school students. Methods: In Gansu, Guangdong, Guangxi, Guizhou, Liaoning, Shandong, Shanxi and other provinces, 1 prefecture was selected, and a number of primary schools were selected from each region. All the students in the class were selected as the object of this survey. A total of 8 365 middle school students were examined for their eyesight, and the data of general population economic indicators and natural environment indicators were obtained through the statistical yearbook of various provinces and cities. The influencing factors of primary school students myopia were analyzed by chi-square test and multivariate Logistic regression. Results: Univariate analysis showed that different provinces and different sex, different nationalities, different grade, parents average salary, sunshine duration, air temperature, altitude, longitude, latitude, different economic zone(χ2=116.22, 18.08, 26.33, 1 059.04, 14.86, 10.28, 16.95, 10.01, 23.15, 29.43, 88.14, P<0.05). Multivariate analysis results showed that gender, grade, sunshine duration, longitude were risk factor for poor vision(OR=1.31, 1.71, 1.45, 1.54, P<0.05); Economic zone and parents salary were protective factors for poor eyesight of students (OR=0.65, 0.86, P<0.05). Conclusion Myopia of primary school students is affected by a variety of factors, economic and social factors and natural environmental factors have an impact on the screening.