Objective
To investigate the effect of liver X receptor α (LXRα) gene silencing on lipid metabolism-related genes in HepG2.2.15 cells.

Methods
HepG2.2.15 cells were divided into blank control group (without transfection), negative control group (transfected with HK plasmid), and shLXRα group(transfected with shLXRα plasmid). The shLXRα plasmids carrying LXRα gene were constructed and were used to transfect HepG2.2.15 cells using PolyJetTM reagent. Green fluorescent protein and LXRα protein expression were examined by fluorescence microscope and Western blotting analysis 24-96 h after transfection, so as to identify the best interference time.

Then cells were treated with agonist T0901317 for 24 h or 48 h; and the content of triglyceride (TG) was observed to detect the degree of steatosis by biochemical assay. The expression of sterol regulatory element binding protein-1c (SREBP-1c) mRNA was detected by RT-PCR and the expression of hepatitis B virus X (HBx) protein and fatty acid synthase (FAS) protein was tested by Western blotting analysis.

Results
The shLXRα plasmid was constructed and transfected into HepG2.2.15 cells successfully. Compared with blank and negative control groups, LXRα protein was markedly decreased in the shLXRα group, with the lowest level found at 48-72 h after transfection (P＜0.01).

After cells were stimulated with T0901317, HBx and FAS protein expression, the content of TG, and SREBP-1c mRNA expression gradually increased with the prolongation of stimulation period, and there was no significant difference in HBx expression at the same time point between different groups. FAS protein, TG contents, and SREBP-1c mRNA in shLXRα group were significantly lower than those in the other two groups (P<0.01).

Conclusion
HBx can regulate lipid metabolism through LXRα/SREBP-1c/FAS pathway.

Keywords:liver X receptor α,hepatitis B virus X gene,lipid metabolism disorders,RNA interference
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Objective
To investigate a new method for creating experimental model of noninvasive fungal sinusitis in rabbits.

Methods
Twenty-five New Zealand white rabbits(50 sides)were divided into seven groups: control group (n=6 sides),sham-operation group (n=6 sides),mucosal-scratched group (n=6 sides),ostia-blocked group(n=6 sides), Aspergillus fumigatus inoculated group(n=8 sides), mucosal-scratched plus Aspergillus fumigatus inoculated group(n=9 sides),and ostia-blocked plus Aspergillus fumigatus inoculated group(n=9 sides).

The experiment was divided according to whether the maxillary sinus mucosa was wounded or not and whether the natural ostia were blocked or not. On days 14 and 28 after Aspergillus fumigatus inoculation, histopathologic and mycological examinations of both maxillary cavities and mucous membrane were performed to confirm the establishment of the model.

Results
Aspergillus fumigatus plus ostial obstruction group had a positive rate of 88.9% (8/9 sides) on day 14 and 77.8% (7/9 sides) on day 28 in fungal culture. Aspergillus fumigatus plus mucosa wound yielded a positive rate of 44.4%(4/9 sides) on day 14 and 22.2%(2/9 sides) on day 28.

Conclusion
Aspergillus fumigatus inoculation combined with an blocked ostium can be used to establish experimental model of noninvasive maxillary fungal sinusitis.

Keywords:sinusitis,fungi,animal models,noninvasive

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Objective
To create transient receptor potential vanilloid 6 (Trpv6) gene knockout mouse model, so as to pave a way for further research of its biological function and its role in bone metabolism in vivo.

Methods
Mouse genomic DNA sequence of Trpv6 gene was obtained from Ensembl database. Trpv6 gene knockout vector (pBR322-MK-Trpv6) was constructed. Trpv6 knockout vector was transferred into the embryonic stem (ES) cells by electroporation and screening of both G418 and Ganciclovoir resistant clones were performed routinely.

The homologous recombined ES cell clones were identified by PCR. The correct homologously recombined ES cells were microinjected into C57BL/6J mouse blastocysts to obtain chimera mouse. Male mice with a chimera rate of 50% were mated with C57BL/6J female mice; the offsprings with gray fur were obtained, which were identified as heterozygote mice by PCR. Heterozygote mice were intercrossed to generate homozygote mice.

Results
Targeting vector pBR322-MK-Trpv6 were successfully constructed. A total of 24 correct homologously recombined clones were gained after electroporation. The efficiency of homologous recombination was 25%. Four male mice with a chimera rate of more than 50% were acquired after homologously recombined clones through microinjection.

After the chimera mice were mated with C57BL/6J mice, 57 grey-fur mice originated from ES cell were gained, including 17 (29.8%) with heterozygous genotype. Heterozygote mice were intercrossed to generate homozygote mice. Western blotting analysis showed no Trpv6 protein expression in homozygote mice.

Conclusion
We have successfully established Trpv6 gene knockout mouse model, and there is no embryonic lethality in homologous mutant mice.

Keywords:trpv6 gene,gene knockout mice,bone remodeling

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Objective
To study the effect of tubulin tyrosine ligase like 12 (TTLL12) and nitrotyrosine on the growth of prostate cancer cells.

Methods
TTLL12 and α-tubulin nitrotyrosination (N-tubulin) were detected by Western blotting analysis in normal prostate epithelial cell lines PWR1E and RWPE1, and prostate cancer cell line DU145; the proliferation of cells was observed by MTT assay.

Results
The expression of N-tubulin was significantly lower in DU145 cells than those in PWR1E and RWPE1 cells after treatment with nitrotyrosine (P＜0.05). The proliferation of PWR1E and RWPE1 cells treated with nitrotyrosine (cultured with 800 μmol/L nitrotyrosine) were significantly inhibited compared with the control (cultured with 800 μmol/L hydrochloric acid, P<0.05), while there was no visible change in the proliferation of DU145 cells.

N-tubulin level was significantly higher in the experimental group (transfected with siTTLL-12) than the control group (transfected with siControl) after effective silence of TTLL12 (P＜0.05); the proliferation of DU145 cells treated with nitrotyrosine in the experimental group was significantly inhibited (P<0.05),while there was no visible change in cell proliferation in the control group (P>0.05).

Conclusion
TTLL12 allows an escape of prostate cancer cells from the treatment of nitrotyrosine,leading to the escape of prostate cancer cells from the body’s surveillance system and acquirement of their abnormal proliferation.

Keywords:tubulin tyrosine ligase like 12,nitrotyrosine, prostatic neoplasms

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