Objective To construct the recombinant RIP3 over-expressed plasmids and transfect them in breast cancer MCF7 cells, and identify the expression and localization of fusion protein, as well as its effect on the death way of MCF7 cells. Methods The expression levels of RIP3 mRNA in four breast cancer cell lines and normal mammary epithelial cells were detected by reverse transcription polymerase chain reaction (RT-PCR). The RIP 3 coding sequence was amplified by polymerase chain reaction and subcloned into mCherry vector to construct recombinant plasmids. The plasmids were transfected into MCF7 cells by lentivirus after DNA sequencing, then screened by basticidin (4 mg/L) for 1 week. The efficiencies of RIP3 expression were validated by Western blotting assay. The death way of mCherry-RIP3-MCF7 cells was observed under the treatment of TNF-αand Z-VAD-FMK. Results The lowest expression of RIP3 mRNA was found in MCF7 cells. The sequencing results validated the well recombinant plasmids. The expression of mCherry-RIP3 fusion pro-tein with a molecular weight of 85 ku was detected by Western blot assay. The mCherry-RIP3 expression enhanced the sensi-tivity of MCF7 cells to TNF-αand Z-VAD-FMK induced cell death. Conclusion The recombinant RIP3 over-expressed plasmids were successfully constructed, and the stable MCF7 cells with ectopic RIP3 transfection were obtained. The mCher-ry-RIP3 fusion protein was expressed in the cytoplasm and was conformed to mediate TNF-αinduced necroptosis.

Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.

Objective • To investigate the effect of 500 kHz pulsed ultrasound combined with SonoVue microbubbles on the vascular permeability of hind limbs in rats and the permeability dynamics for the duration. Methods • Eighty male Sprague Dawley rats were randomly divided into eight groups (n=10), including control group, only microbubble group (MB group), and microbubble combined with ultrasound groups (MB+US group). And MB+US groups classified into time subgroups according to the interval from the endpoint of ultrasound exposure to the injection of Evans blue (EB), included 2.5 min group, 5 min group, 10 min group, 20 min group, 30 min group and 60 min group. The change of vascular permeability was quantitively analyzed by detecting the content of EB after ultrasound irradiation. Meanwhile, four rats in each group were randomly selected. Two of them were performed hematoxylin-eosin (H-E) staining; and the rest of them underlied transmission electron microscope analysis. Results • The results of EB content detected from muscle samples demonstrated that the exposure of ultrasound in conjunction with microbubbles could alter the vascular permeability of hind limbs of rats. The optimal time window for extravasation was within 30 min after ultrasound exposure. And the permeability could return to the normal status 60 min after ultrasound irradiation. There was no apparent damage on arteries and muscles in the slices with H-E staining. From the observation of transmission electron microscope, the tight junctions between endothelial cells on the arterial wall were significantly widened in MB+US groups. Conclusion • The application of ultrasound with the frequency of 500 kHz combined with SonoVue microbubbles can reversibly change the vascular permeability of hind limbs in rats without obvious damage to the tissues.

Dihydrofolate reductase (DHFR) is a well-known key target in the treatment of tumors, bacterial infections, and parasitic infections; and it plays a critical role in the biosynthesis of cellular DNA. DHFR inhibitors interfere with one-carbon metabolism by inhibiting substrate binding to DHFR, thereby inhibiting cell proliferation. Research on DHFR inhibitors has continued since the 1940s. To date, a variety of DHFR inhibitors have come into the market, primarily used for anti-tumor, antibacterial, antiparasitic, and anti-inflammatory therapy. This review summarizes the research progress of DHFR inhibitors with antitumor or antibacterial effects in recent years based on the classification of single-target and dual-target and looks forward to the opportunities and challenges faced by the work in this field.

The virtual colonoscopy, virtual flattening and virtual splitting method are enhanced by the GPGPU model. The novel virtual eversion method is integrated for fast polyp detection. The experimental result showed that the system and various visualization methods can represent the colon inner-surface clearly and exactly, supporting real-time man-machine interaction. The proposed system is promising in human gastrointestinal cancer and polyp inspection.
Colonic Polyps ; diagnosis ; Colonography, Computed Tomographic ; instrumentation ; methods ; Humans ; Imaging, Three-Dimensional ; Software Design ; Tomography, X-Ray Computed

OBJECTIVETo observe the effective of the injectable artificial bone combined with plate fixation for reconstructing the collapse tibial plateau fracture.

METHODSFrom June 2005 to January 2008,21 cases of collapse tibial plateau fracture of type Schatzker II, III were treated by injectable calcium sulfate bone substitute combined with supportive plate reconstruction including 16 males and 5 females with an average age of 35.3 years ranging from 27 to 62 years. The disease course was from 3 to 7 days (means 4 days). According to Schatzker classification, there were 17 cases of type II, 4 of type III. All patients preoperatively underwent radiography, CT scanning and three-dimensional reconstruction in order to accurately understand the extent of fracture and fracture collapse and the shattered fragments of the flip direction. All the fracture with collapse > 3 mm, without joint degeneration were selcected for surgical treatment. The knee joint function after fracture healing and recovery were evaluated by Lysholm scoring.

RESULTSAll patients were followed-up for from 6 months to 2.5 years (means 1.5 years). The X-ray films and features of all fractures showed anatomic reduction or near anatomic reduction, except one case of grade II severe comminuted fracture occurred a high degree of loss and platform reset ineffective after 6 months. The Lysholm scoring of knee function showed that the mean score was (88.3 +/- 5.2). The results were excellent in 12 cases,good in 7 cases, fair in 2 cases.

CONCLUSIONMinimally invasive injectable calcium sulfate bone combined with plate fixation for reconstructing the collapse tibial plateau fracture of type II, III can effectively prevent the further loss after reduction, to improve the long-term results. Minimally invasive injectable calcium sulfate as an artificial bone substitute materials has good prospects for clinical application.

Adult ; Bone Plates ; Bone Substitutes ; administration & dosage ; Calcium Phosphates ; administration & dosage ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Injections ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Tibial Fractures ; surgery

Spinal cord injury(SCI)can lead to varying degrees of somatic and autonomic dysfunction and seriously affect the quali-ty of life of patients.The neurobiology and function promotion effects of brain-derived neurotrophic factor(BDNF)after SCI have been extensively studied,including neuroprotection,neuroplasticity and process regulation of neuroinflamma-tion,etc.,as well as functional recovery.However,the adverse effects of BDNF after SCI,such as neuropathic pain and spasticity,were also studied.

OBJECTIVE: To investigate the expression of fibroblast growth factor receptor 2 (FGFR2) in clear cell renal cell carcinoma (ccRCC; or kidney renal clear cell carcinoma, KIRC), to analyze the relationship between the expression of FGFR2 and the clinical pathological features and prognosis of ccRCC, to study the relationship between the expression of FGFR2 and other molecules, and to explore its role in the development of ccRCC. METHODS: Gene expressional and clinical information of ccRCC patients were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus(GEO) database. Next, the data were transformed and collated. In the study, 104 clinical ccRCC samples and corresponding paracancerous normal tissue samples were collected from Department of Urology, Peking University First Hospital. Immunohistochemistry (IHC) was performed and the staining results were scored, so as to compare the expression of FGFR2 in ccRCC and paracancerous normal tissues. Besides, quantify real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression level of FGFR2 in normal renal epithelial cell lines (293) and ccRCC cell lines (786-O, 769-P, OSRC-2, Caki-1, ACHN, and A498). In addition, the relationship between FGFR2 expression and clinical pathological characteristics (including TNM staging and pathological grading) and survival prognosis in ccRCC patients was further analyzed. Furthermore, the relationship between FGFR2 expression and B cells, T cells, natural killer (NK) cells and neutrophil infiltration in the ccRCC patients was analyzed, and the Biological General Repository for Interactionh Datasets (BioGRID) was used to builds protein-protein interaction (PPI) networks to study molecules that interacted with the FGFR2 protein. RESULTS: In the TCGA database, the expression of FGFR2 was down-regulated in ccRCC tissue samples compared with normal tissue samples, and the expression in the GEO database also showed this differences. Furthermore, FGFR2 expression was downregulated in ccRCC clinical samples and ccRCC cell lines, compared with corresponding paracancerous normal tissue or normal renal epithelial cell lines. In addition, FGFR2 high expression was associated with earlier, lower-level ccRCC and was associated with a better prognosis in the patients with ccRCC. Moreover, FGFR2 expression was not significantly related to B cells, T cells, NK cells and neutrophil infiltration, and the PPI network showed that FGFR2 protein interacted with certain molecules. CONCLUSION Our work sheds light on the potential role of FGFR2 in the development of ccRCC, suggesting that FGFR2 may serve as a prognostic marker and potential therapeutic target for patients with ccRCC.
Biomarkers, Tumor/analysis* ; Carcinoma, Renal Cell/pathology* ; Humans ; Kidney Neoplasms/pathology* ; Prognosis ; Receptor, Fibroblast Growth Factor, Type 2/genetics*

OBJECTIVETo evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis (PGD) in couples at risk of having child with beta-thalassemia.

METHODSSingle cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia.

RESULTSIn six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out(ADO) rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination.

CONCLUSIONThis is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene(HumTHO1) could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.

Adult ; Female ; Humans ; Male ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis ; methods ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVETo establish a monoclone cell line of squamous cell carcinoma (SCC) in rat buccal mucosa and to study its biological characteristics.

METHODSSCC in rat oral mucosa was induced by adding 4-nitroquinoline-1-oxide (4NQO) into the SD rats' drinking water, and the cancer cells were then cultured to obtain mixed cells in vitro. The mixed tumor cells were purified by mono cell cloning method. The biological characteristics of the cells were studied by microscope and electronic microscope observation, chromosome analysis, Methyl thiazolyl tetrazolium (MTT) test, flow cytometry assay and immunohistochemistry staining. Hypodermic inoculations of the cells in nude mice and injection of the cells by nude mice tail veins were performed to observe the tumor formation and long distance metastasis.

RESULTSThe morphology proved that the cell line was squamous cell carcinoma cells, which were cultured from one cell. The population doubling time for passage 65 cells was 25.44 hours. The cells in S-phase accounted for 20.13% of the cell cycle. The chromosome modal number was 84. All the cells expressed the proteins of cytokeratin and vimentin. The xenograft rate and the tumor metastatic rate to the lung were 100% in nu/nu BALB/C mice, but the homograft rate was zero in SD Rats.

CONCLUSIONSRca-B was a typical oral squamous cell carcinoma cell line derived from Sprague-Dawley rat buccal mucosa carcinoma, and the cell line has high metastatic potential and its biological characteristics were well ascertained.

4-Nitroquinoline-1-oxide ; toxicity ; Animals ; Carcinoma, Squamous Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Clone Cells ; pathology ; Female ; Mice ; Mice, Nude ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; pathology ; Rats ; Rats, Sprague-Dawley