Objective To study the relationship between the inherit gene and the pharmic efficacy by observing the apolipoprotein E polymorphism and the nylestroil effect in lowering serum lipids in postmenopausal women. Methods Ninety-four postmenopausal women were divided into three groups according to their types of the apoE gene: E2, E3 and E4. Thirty-six women who received nylestroil therapy were compared with controls, 58 women who had not received the therapy, in lipids, apolipoprotein E and lipoprotein(a)〔LP(a)〕. Results Compared with the control groups, the treatment groups showed an increase in HDL-C (high-density lipoprotein) and decrease in LDL-C(low-density lipoprotein) and LP(a). Though Group E2 showed an increase in the HDL-C and TG and a decrease in the LDC-C and LP(a), no significance was shown compared with the controls. Group E3 was increased in the HDL-C(2.7?0.9)mmol/L and decreased in the LDL-C (3.2?1.0)mmol/L and Lp(a) (0.4?0.2)g/L, and the differences were significant in HDL-C(1.7?0.5)mmol/L and Lp(a) (0.5?0.3)g/L compared with the controls. Group E4 was increased in the HDL-C (2.6?1.0)mmol/L and decreased in the LDL-C(2.5?1.6)mmol/L, TC and LP (a), and the differences were statistically significant(P

Objective:To study the possible immune escape mechanisms of HCoV-OC43 from human dendritic cells(DC).Methods:HCoV-OC43 was isolated from clinical specimen using BSC-1 cells and identified by Real-time PCR,and the cytopathic effect was observed by phase contrast microscope.DCs were induced in vivo using hu-GM-CSF and IL-4 cytokines,and after 7 days of differentiation,DCs were infected by HCoV-OC43.The morphology of HCoV-OC43 infected DC was observed by transmission electron microscope,and the cytokines related to DC functions were detected by Real-time PCR after infection.DC proportion and function related co-stimulatory molecules were analyzed by flow cytometry.Results:In vitro HCoV-OC43 infected human DC model was successfully built.HCoV-OC43 can infect DC and generate immune response of DC in vitro,but no virus nucleonic acid could be detected in culture supernatant.The DC expression of IFN-α,IFN-β,CCL3 and CCL5 were significant decreased when infected with HCoV-OC43,but the expression of costimulatory molecules including HLA-DR,CD1c and CD86 were not affected by HCoV-OC43 infection.Conclusion:Human DC could be infected by HCoV-OC43 and generate immune response,but could not produce progeny virus.HCoV-OC43 may escape from immune response by suppressing the expression of IFN-α and other inflammatory cytokines and chemokines in DC.

Objective To investigate the precision and stability of optical surface imaging (OSI)system Catalyst in guiding radiotherapy positioning.Methods A total of 52 patients with five different tumor sites who underwent cone-beam computed tomography (CBCT)-guided radiotherapy were recruited in this investigation.For the first treatment fraction,the setup error was recorded as C after online CBCT correction,and the surface images of patients taken by Catalyst were set as the reference images Cref.For the following treatment fraction,patients were pre-corrected according to the Catalyst Cref image with the acceptable errors within 2 mm/ 2,and the pre-corrected errors were recorded as C1.Then,after online CBCT correction,the setup errors were recorded as C.The errors between post-corrected Catalyst surface image and Cref image were recorded as C2.For each treatment fraction,the difference between Catalyst correction errors C1 and CBCT corrected errors C was recorded as d1,and the difference between the post-corrected Catalyst errors C2 and Cref image was recorded as d2.d3=d1-d2.The values of d1 and d3 in the 6 dimensions were analyzed using single sample t-test.The correlation between C-C1 and d1-d2 was statistically analyzed by Pearson correlation analysis.Results The mean value of d1 and d3 for 52 patients were within 2 mm/2 °.CBCT-C1 and d1-d2 were both significantly correlated (R =3,7,P=0.00,0.01).Conclusions OSI system yield high accuracy and stability in radiotherapy positioning,which is of certain significance in radiotherapy positioning for cancer patients.

In order to study the molecular mechanism and physiological significance of the interaction between PGRN and Rev-erbβ, the PGRN gene in HEK293 (Rev-erbβ-/-) marked as C3-6 cell lines was knocked out by CRISPR/Cas9 system to generate the Rev-erbβ and PGRN double genes knockout HEK293 cell lines. First, four sgRNAs were designed for PGRN gene, and PGRN sgRNA2 and sgRNA3 with the higher activity were used to construct the Lentiviral vector, pLenti/CMV-Loxp-Cas9-sgRNA2-U6-sgRNA3-U6-Loxp-EF1α-Puro. Then, the lentivirus vector carrying Cas9 and double PGRN sgRNA were used to infect HEK293 C3-6 cells. Through drug screening, cloning and sequencing, we obtained the monoclonal HEK293 (Rev-erbβ-/-; PGRN-/-) marked as C3-6/23 cell lines. Using qRT-PCR and Western blotting, we detected PGRN mRNA and protein expression in C3-6/23 cell lines. Finally, genetic complementation was used to study the effect of PGRN-mediated Rev-erbβ on the regulation of the target gene promoter transcriptional activity in the C3-6/23 cell lines. In HEK293 C3-6/23 cell lines, the two DNA chains of PGRN gene were both deletion mutagenesis, and the expression mRNA and protein of PGRN did not reach the detection level. At the same time, the interaction between PGRN and Rev-erbβ enhanced the regulation of Rev-erbβ on the transcription of target gene promoter in the cell lines. Using CRISPR/Cas9 system, we successfully constructed the double knockout HEK293 (Rev-erbβ-/-; PGRN-/-) monoclonal cell lines. The study found that PGRN could affect Rev-erbβ on the regulation of target gene promoter transcription in the C3-6/23 cell lines; however, the mechanism of PGRN involvement in mediating Rev-erbβ in transcriptional regulation remains to be further studied.

Objective To investigate the changes of autophagy in rat lung tissues after acute spinal cord injury (ASCI) and the possible mechanisms.Methods Sixty-three female SD rats were divided into sham operation group (n =21) and spinal cord injury group (n =42),according to the random number table.The modified Allen method with the impact energy of 10 × 25 g · mm at the T10 vertebra was used for preparation of ASCI models.The rats were sacrificed at 6,12,24,48,72 hours,1 and 2 weeks after injury.Lung tissue damage and apoptosis were detected by HE staining and TUNEL method.The changes of autophagy and expressions of LC3-Ⅱ,P62,Beclin-1,interleukin (IL)-17A and Bcl-2 in lung were detected by Western blot and immunofluorescence.Results The pulmonary alveoli maintained normal structure in sham operation group,with no inflammation or pulmonary hemorrhage.Slight lung tissue damages were observed in spinal cord injury group at 12 h postinjury.Alveolar stroma widening,inflammatory infiltration,hemorrhage,and alveolar collapse became ingravescent at 24-72 hours postinjury.Numbers of apoptotic cells in spinal cord injury group were 551.22 ± 135.94,905.11 ±92.64,and 141.78 ± 30.86 respectively at 24,72 hours and 1 week postinjury,and were significantly increased at 24 and 72 hours postinjury,compared with sham operation group (P ＜ 0.05).Expression of LC3-II in spinal cord injury group was increased at 24-72 hours postinjury,compared with sham operation group (P ＜ 0.05).Expression of P62 in spinal cord injury group was up regulated at 24-72 hours postinjury,compared with sham operation group (P ＜ 0.05).Expression of Beclin-1 in spinal cord injury group was increased at 24 h postinjury and then dropped at 48-72 hours,compared with sham operation group (P ＜ 0.05).Expression of IL-17A in spinal cord injury group was increased at 24-48 hours,compared with sham operation group (P ＜ 0.05).Expression of Bcl-2 in spinal cord injury group was increased from 24 hours to 72 hours,compared with sham operation group (P ＜ 0.05).Conclusion Autophagosome formation is increased and accumulated in the lung tissues after ASCI,which might be related to the increased interaction between Beclin-1 and Bcl-2 because of the up regulation of Bcl-2 by IL 17A,ultimately leading to the inhibition of autophagy.

Objective: To investigate the molecular evolution characteristics of human coronavirus (HCoV) subtypes in patients with fever and respiratory tract infection in Guangzhou from 2010 to 2012. Methods: Partial fragments of NP, RdRp and S genes of HCoV-OC43, HCoV-229E and HCoV-NL63 positive samples were amplified by RT-PCR and sequencing. Bioinformatics software, including Bio-edit, Mega4.0 and Clustal1.83 were used for comparison and analysis of NP, RDRp and S gene sequences. Molecular evolutionary tree of different gene regions of HCoV-OC43, HCoV-229E and HCoV-NL63 were built. Results: No remarkable variation or recombinant strain of HCoV-OC43, HCoV-229E and HCoV-NL63 was found in Guangzhou during 2010—2012. The HCoV-OC43 substrains were genetically closest to the strains found in Belgium and Hong Kong (GenBank accession number JN129834 and AY903460). HCoV-229E substrains were genetically closest to those found in Amsterdam (GenBank accession number JX503060) and HCoV-NL63 most genetically close to those in Amsterdam and Beijing (GenBank accession number JX104161 and DQ445911). The NP and RDRp genes of all subtypes were highly conserved, while S gene was more variable. Conclusions There were at least 3 substrains of HCoV-OC43, HCoV-229E and HCoV-NL63 epidemic in Guangzhou during 2010—2012, and no remarkable variation or recombinant viral strain was found. The NP and RDRp genes of all subtypes were highly conserved and can be used in virus detection, while S gene was more variable and suitable for phylogenetic and variation study.