Epigenetics change has been considered to be the most promising new strategy for disease control and prevention. TCM regulates gene expression through epigenetics, participating in pathological and physiological process including cell apoptosis, proliferation, differentiation, cell cycle regulation, immunity, inflammation, and metabolism. This article reviewed the application of DNA methylation, histone modification and the miRNA regulation in TCM research.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

BACKGROUND: Strontium-doped calcium polyphosphate (SCPP) is a new type of bone repair materials with good biocompatibility and controlled degradation. The preliminary studies of our group indicate their role in promoting angiogenesis,but its mechanism is unclear.OBJECTIVE: By co-culturing osteoblasts ROS17/2.8 with SCPP in vitro to observe cell proliferation and the secretion of vascular endothelial growth factor (VEGF).DESIGN, TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2008 to June 2009.MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 0%, 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared. ROS17/2.8 osteoblastic cell strain was provided by Laboratory of Transplantation Immunity and Transplantation Engineering, West China Hospital, Sichuan University.METHODS: ①Preparation of cell scaffold complexes: The materials were placed in 24-well plates, then 300 μL cell suspension with a concentration of 2×10~7 cells/Lwas inoculated into each hole. These complexes were cultured for 14 days and the liquid was changed every two days. ②These complexes were measured by MTT assay to observe the proliferation of osteoblasts on the 1~(st), 3~(rd), 5~(th), 7~(th), 10~(th) and 14~(th) days, respectively. ③ The centrifugal supernatant of the complex cultured for seven days was measured by ELISA assay to check the secretion of VEGF.MAIN OUTCOME MEASURES: The proliferation of osteoblastic cells on SCPP and CPP was observed. The amount of VEGF protein secreting from osteoblastic cells was detected.RESULTS: The results of MTT showed that, compared with the CPP group, SCPP groups could promote the proliferation of osteoblasts, and 8% SCPP group was the best; ELISA results showed that, compared with the CPP group, SCPP groups could increase the amount of VEGF protein secretion, of which the promoting role of 8% SCPP was the most obvious (P < 0.05).CONCLUSION: When cultured with osteoblasts, SCPP can promote cell proliferation, and can significantly increase the secretion of VEGF; moreover, 8% SCPP is the best, which reveals a certain mechanism of its promoting angiogenesis.

Chondrosarcoma is extremely rare in maxillofacial soft tissue. A case diagnosed as well-differentiated chon- drosarcoma in the left parotid region was reported. The clinic pathological features, diagnosis, treatment, and prognosis were discussed with the literature review.
Bone Neoplasms ; Chondrosarcoma ; Humans ; Parotid Neoplasms ; Parotid Region

Ossifying fibromyxoid tumor is an uncommon neoplasm with uncertain histogenesis. This tumor is usually characterized by a small, painless mass in the subcutaneous tissue or limb muscles. In this case, an ossifying fibromyxoid tumor of the mandible was reported, and relevant literature was reviewed.
Fibroma ; Fibroma, Ossifying ; Humans ; Mandible

Tongue osteocartilaginous choristoma is the disease that there are well-developed bone and cartilage in the tongue. This article reported a case of tongue osteocartilaginous choristoma in the oral-cavity,which is rare.
Cartilage ; Choristoma ; Humans ; Tongue Diseases

Objective To investigate the role of transcriptional-coactivator with PDZ-binding motif( TAZ) in genistein-induced osteoblastogenic differentiation of mouse bone marrow-derived mesenchymal stem cells ( BMSCs) .Methods Mouse BMSCs were cultured in phenol red-freeα-MEM containing osteogenic supplements for inducing osteogenic differentiation.BMSCs were transfected with siRNA-TAZ and treated with genistein.The temporal sequence of osteoblastic differentiation in BMSCs cultures was assayed by measuring alkaline phosphatase activity (ALP) and calcium deposition.The mRNA expression of bone sialoprotein ( BSP) and osteocalcin ( OC) were detected by reverse transcription-polymerase chain reaction(RT-PCR).The binding interaction between TAZ and cbfa1 was identified by co-immunoprecipitation.Results TAZ expression was detected during the induction of osteogenic differentiation, the ALP activity and calcium deposition were significantly decreased in BMSCs which were transfected with siRNA-TAZ.Genistein(0.01-1 μmol/L) exhibited a dose-dependent effect on TAZ expression in mouse BMSCs cultures.Treatment with genistein ( 1 μmol/L ) resulted in increased ALP avtivity and calcium deposition of BMSC cultures as function of time.Genistein(1μmol/L) also promoted the nuclear localization of TAZ and augmented the interaction between TAZ and cbfa1, and by which upregulated cbfa1-mediated gene expression such as BSP and OC.However, the ALP avtivity and calcium deposition, as well as the expression of BSP and OC were not promoted by genistein in BMSCs transfected with siRNA-TAZ.Conclusion These data suggest that the TAZ plays an important role in genistein-induced osteoblastic differentiation of mouse BMSCs cultures.

OBJECTIVETo analyze the rules for acupoint selection of acupuncture and moxibustion in domestic clinical treatment of perimenopausal syndrome based on data mining technology in modern times.

METHODSThe relevant literature were retrieved from Chinese Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI) and Wanfang database on this disease treated with clinical acupuncture and moxibustion in China from 1978 to 2013. The database of acupuncture-moxibustion prescription was set up. The relevant regulations of data mining technology were used to analyze the rules for acupoint selection.

RESULTSTotally, 211 papers, 254 acupuncture-moxibustion prescriptions and 130 acupoints were included. The total frequency of acupoints application was 2193 times, with 14 meridians involved. The utilization of the acupoints in the lower limbs and on the back were 33.0% (723/2193) and 23.8% (521/2193) and those of yin and yang meridians were 51.8% (1136/2193) and 44.0% (965/2193), respectively. The utilization of the specific acupoints accounted for 88.7% (1946/2193).

CONCLUSIONIn clinical treatment of perimenopausal syndrome with acupuncture and moxibustion in modern times, the acupoint selection from involved meridians is the basis, associated with multiple methods of acupoint combination; yin and yang meridians are equally important and the specific acupoints are considered particularly critical in application.

Purpose To investigate the relationship and clinical significance of high-risk human papillomavirus infection and expression of c-jun and c-fos protein in cervical carcinoma.Methods Cervista HR-HPV testing,immunohistochemical SP method were employed to detect the infection of HR-HPV,c-jun and c-fos protein in 70 cases of CSCC,60 cases of cervical intraepithelial neoplasia and 20 cases of chronic cervicitis.The correlation between c-jun and c-fos protein expression and of HR-HPV infection was analyzed.Results Of the 70 cases of CSCC,the positive HR-HPV was 69 cases,the positive rate in HR-HPV in A9 group was the highest (85％,59/69).The positive expression of c-jun and c-fos were 80％ (56/70),85.7％ (60/70) in 70 cases CSCC,70％ (21/30) and 70％ (21/30) in 30 cases of cervical high grade intraepithelial neoplasia,and 20％ (6/30),23.3％ (7/30) in 30 cases cervical low grade intraepithelial neoplasia,but the positive expression rates of c-jun and c-fos in the 20 cases of chronic cervicitis were 0.Expression of c-jun and c-fos in cervical cancer group was higher than that in the chronic cervicitis group and low grade intraepithelial neoplasia (P ＜ 0.05).The expression of c-jun and c-fos was statistically significant in different groups of clinical stage and pathologic grading in the CSCC (P ＜ 0.05).Additionally,the expression of c-jun and c-fos was positively correlated with the infection of HR-HPV in CSCC (P ＜ 0.01).Conclusion The infection of HR-HPV has significant subtype-specific and has a positive correlation with the expression of c-jun and c-fos in CSCC,which suggests that AP-1 pathway activation after HRHPV infection may be associated with the occurrence and development of cervical carcinoma.