Objective:To construct the retroviral vector containing HBx gene so as to investigate the effects of HBx gene in the pathogenesis of HBV-dependent hepatic cell carcinoma(HCC)and further construct animal models of hepatic cell carcinoma.Methods:HBx gene was amplified from the HBx adenovirus Plasmid by polymerase chain reaction(PCR)technique whose primer was designed according to the announced sequences of HBx gene in gene bank,then cut by two endonucleases and directionally cloned into the pseb-hus vector and analyzed by gene sequencing analyzer,the correct constructed vectors were transfected into L02 cell line.At last,HBx protein was detected by Western Blotting.Results:Through PCR,endonuclease cutting and gene sequencing,the target gene was verified into be correctly cloned to retroviral plasmid pSEB-HUS and the high expression of green fluorescence protein(GFP)in L02 cell line was observed under fluorescent microscope.The expression of HBx protein was also observed by Western Blotting.Conclusion:Recombinant retroviral vector expressing HBx protein was successfully constructed.

OBJECTIVE: To investigate the protective effect of berberine on CCl4-induced acute liver injury in mice.METHODS: Kunming mice were included and equally divided into 6 groups: normal group,model group,berberine groups(40,20,10 mg?kg-1).After intragastric administration of corresponding drugs for 5 consecutive days,the mice were intraperitoneally injected with CCl4(10 mL?kg-1) to establish liver injury mice model.Alanine transaminase(ALT) and aspartate transaminase(AST) in serum,activities of superoxidase(SOD) and malondialdehyde(MDA) content in liver homogenate were measured.RESULTS: In Berberine-treated groups compared the model group,serum ALT level and AST activity decreased significantly(P

According to the registered product standards specification of medical devices, combined with the standard reviewing work, the common problems of standards during medical devices registration were analyzed and corresponding suggestions were proposed to standardize the standard of the registered product, accelerate the standardization and promote the industry standardized.
Device Approval ; Equipment and Supplies ; standards

For the existing measurement methods of residual voltage which can't turn the power off at peak voltage exactly and simultaneously display waveforms, a new residual voltage detection method is put forward in this paper. First, the zero point of the power supply is detected with zero cross detection circuit and is inputted to a single-chip microcomputer in the form of pulse signal. Secend, when the zero point delays to the peak voltage, the single-chip microcomputer sends control signal to power off the relay. At last, the waveform of the residual voltage is displayed on a principal computer or oscilloscope. The experimental results show that the device designed in this paper can turn the power off at peak voltage and is able to accurately display the voltage waveform immediately after power off and the standard deviation of the residual voltage is less than 0.2 V at exactly one second and later.
Electric Power Supplies ; Equipment Design ; Microcomputers ; Signal Processing, Computer-Assisted

Objective To evaluate the relationship between the risk of breast cancer and abortions among nulliparous women. Methods Searched the data of Cochrane Library and PubMed before June 2014 to identify potentially studies which involved the relationship between the risk of breast cancer and abortions among nulliparous women. Data was extracted by two independent authors from each study. STATA software was used for statistical analysis. Calculated the pooled RR and 95% CI as the assessment of the link between abortions and breast cancer in fixed effects models. Results 13 studies were included. The study showed the RR and 95%CI of the relationship between the risk of breast cancer and abortions was 0. 98[0. 89,1. 08],P>0. 05 in nulliparous women, and the number of abortions was not associated with the risk of breast cancer. The RR and 95%CI of the relationship between the risk of breast cancer and induced abortions or spontaneous abor-tions were 0. 96[0. 88,1. 04],1. 01[0. 88,1. 14], respectively. Conclusion There is no correlation between breast cancer and abortions a-mong the nulliparous women, and the risk of breast cancer would not increas as the number of abortions increase.

This study aimed to investigate the effects of fusion proteins GnRH-GRP(G3G6)and HSP65-STEAP1(HST1)on dendritic cells(DC)and the sensitization of DCs to B16F10 melanoma. The fusion proteins G3G6 and HST1 were obtained using the previous engineering strains in our laboratory. Group by unsensitized DC(US-DC), the G3G6 fusion protein sensitized DC, the HST1 fusion protein sensitized DC(HST1-DC)and the combined sensitized DC(GH-DC), the mouse bone marrow-derived DCs were sensitized with fusion protein to obtain the fusion protein sensitized DC vaccines. B16F10 melanoma cells were transplanted into C57BL/6J male mice to construct a melanoma model(1×106 cells per mouse), and DC vaccine was injected for treatment. The antitumor efficacy of DC vaccine was explored by in vitro and in vivo experiments. Flow cytometry analysis showed that the fusion protein can effectively stimulate DC into differentiation and maturation; in the animal experiment, the inhibition rate of melanoma treated with G3G6-DC was 35. 75%, that of HST1-DC group and combination group were 34. 03% and 55. 74%. It was initially proved that both G3G6-DC and HST1-DC can effectively inhibit the growth of transplanted tumors of melanoma B16F10 cells in mice, and the combination therapy is superior to the single therapy.

Objective To investgate role of TLR2 in the activation, the innate immune and inflammation of human platelets. Methods Human washed platelets were separated from healthy people(n=5) and were stimulated with different concentrations(1μg/ml, 5μg/ml, 10μg/ml) of TLR2 agonistPam3CSK4(a synthetic bacterial lipoproteins). Then the platelet aggregation rate, the expression of CD62p and TLR2 on the platelet surface were measured. Results The platelet aggregation rate were (28.32±5.67)%, (52.56±8.54)% and (76.24±11.23)%, respectively, at concentration of 1μg/ml, 5μg/mland 10μg/ml of Pam3CSK4, more than (12.83±2.43)% at 0μg/ml of it. In addition, the expression of CD62p were (18.45±2.66)%, (22.45±2.04)%, (29.53±4.08)%, respectively at above concentration of Pam3 CSK4, more than (11.20±1.67)% of CD62p at control group(P<0.01). The expression of TLR2 was not significantly increased at a lower concentration of Pam3CSK4(1μg/ml) with (16.85±6.10)% compared with(10.81±3.99)% at the control group. However, it were (21.15±9.90)% and (22.52±9.26)%, respectively, at a higher concentration(5μg/ml, 10μg/ml)more than(10.81±3.99)% at the control group(P<0.05). Conclusion Pam3CSK4 induce aggregation, activation and the up-regulation of TLR2 of platelet by stimulating TLR2 receptor of it. Thereafter, TLR2 play an important role in the innate immune of platelet.

Objective To investigate the relationship between antipituitary antibodies (APAs) and hypopituiarism following traumatic brain injury (TBI),as well as the severity of brain injury.Methods The study included 73 patients who suffered TBI 9 to 12 months ago and were diagnosed with hypopituitarism during the follow-up.Based on their Glasgow Coma Scale (GCS) on admission they were categorized into three groups:A (3-8),B (9-12) and C (13-15).Levels of plasma pituitary hormones (GH,TT,FT3 and FSH/LH) and APAs were measured in all patients with chemiluminescence assays and ELISA,respectively.Results Patients in group A had higher levels of APAs and lower levels of hormones compared with those in group B(P＜0.001) or group C(P＜0.001),while no significant difference was found between group B and group C for levels of either APAs (P＞0.05) or hormones (P＞0.05).Levels of APAs were negatively correlated with both GH (r=-0.64071,P＜0.001) and GCS (r=-0.50132,P＜0.001).Conclusion The present investigation provides preliminary evidence that APAs may be associated with the development of TBI-induced hypopituiarism.It suggests that the severity of hypopituiarism following TBI could be predicted by measuring the level of APAs.

ObjectiveTo investigate the correlation between the methylation status of HLA class Ⅰ genes(HLA-A, -B and -C) in psoriatic epidermis and disease severity in patients with psoriasis vulgaris. MethodsDNA specimens were obtained from the lesional and nonlesional epidermis of 46 patients with psoriasis vulgaris and from the normal skin of 28 human controls. Methylation specific PCR (MSP) was conducted to detect the methylation status of CpG islands in the promoter region of HLA-A, -B and -C genes. The severity of psoriasis was evaluated by psoriasis area and severity index(PASI) scores. ResultsThe percentage of promoter methylation of HLA-B and HLA-C genes was 4.35％(2/46) and 21.74％(10/46), respectively in nonlesional epidermis, 4.35％ (2/46) and 4.35％ (2/46), respectively in lesional epidermis from these patients. No methylation was observed for the promoter of HLA-A, -B or -C gene in the normal control epidermis or for that of HLA-A gene in the nonlesional or lesional epidermis from the patients. The frequency of HLA-C gene promoter methylation in the nonlesional epidermis was significantly higher than that in the lesional epidermis and control epidermis, but was uncorrelated to the disease severity. No significant difference was observed for the methylation frequency of HLA-A or -B gene promoter among the three groups of specimens. Conclusion Abnormal methylation of HLA-C gene promoter is observed in patients with psoriasis vulgaris.

ObjectiveTo evaluate the relationship between the proliferation of parathyroid cell in rabbit with primary hyperparathyroidism (PHPT) and the bone mineral density (BMD). MethodsEighty adult Chinese rabbits were randomly and equally divided into two groups. The contrast group was fed with normal diet ( Ca ∶ P, 1.0 ∶ 0. 7 ) and the experimental group was fed with high phosphate diet ( Ca ∶ P,1.0∶7.0) to establish the animal model of PHPT. At 3, 4, 5, and 6 months after the diet, bone mineral density of the rabbits was measured by the quantity CT (QCT). Then, the parathyroid and bone of the rabbits were removed for pathological examination. The number of parathyroid cell in PHPT was calculated.Proliferation was determined by immunohistochemistry of proliferation cell nuclear antigen ( PCNA ) and Bcl-2. The t test and Logistic regression was used to analyze the difference of data of two groups. ResultThe number of parathyroid cell in PHPT group was 1.61 times than that in the contrast group[ (673 ± 151 ) HP,(418 ± 25 ) HP,P ＜0. 01]. The rate of PCNA positive-cell was significantly increased in PHPT group than that in contrast group [(50.52 ± 11.62)％o, (26.70 ± 2. 78 )％, P ＜ 0.01], and so was Bcl-2[ (460. 37 ± 190. 05 )‰, (67. 02 ±:4. 38 )％‰,P ＜0. 05]. The value of BMD was significantly decreased in PHPT group than that in contrast group [ ( 152. 5 ± 34. 3 ), ( 188.6 ± 12. 2 ) g/cm3, P ＜ 0. 05]. There was a negative correlation between BMD and PCNA (r ＝ -0. 749, P ＜ 0. 05 ) and between BMD and Bcl-2 (r ＝-0.800, P ＜ 0. 05 ) in PHPT group. ConclusionThe BMD of PHPT is related to the parathyroid cells proliferation which provide a reliable method for early diagnosis of PHPT.