Hypophosphatemia is a common metabolism disease in humans. Fibroblast growth factor 23 (FGF23) inhibits phosphate reabsorption by targeting on the renal tubules. FGF23^C-tail contains 73 amino acids from C-terminus of FGF23, serves as an inhibitor of FGF23, and can increase phosphate reabsorption. Therefore, FGF23^C-tail is an important drug for hypophosphatemia. In this paper, we constructed a fusion protein of FGF23^C-tail with HSA, and investigated the expression of the fusion protein in the Pichia pastoris system. The recombinant gene was constructed by fusion PCR. A high-yield strain was selected by G418 resistance and fermentation yield, and the expression yield was 43.7 mg·L^-1 in flask. In 5 L fermenters, the highest expression yield could reach 265.6 mg·L^-1. FGF23^C-tail-HSA could be used as an inhibitor for FGF23, and could significantly increase blood phosphorus levels in rats. The procedures for care and use of animals were approved by the Ethics Committee of YiChun University. This paper provided a basis research for further studying physiological activity of FGF23^C-tail-HSA.

AIM: To explore the antibacterial activity and underlying mechanism of ursolic acid combined with fusidic acid against Staphylococcus aureus (SA) ATCC29213 and Methicillin-Resistant Staphylococcus aureus (MRSA) ATCC43300 in vitro. METHODS: The minimum inhibitory concentration (MIC) of the combined use of ursolic acid and fusidic acid on SA, MRSA was determined by the micro broth dilution method and the micro checkerboard method, and the partial inhibitory concentration index (FICI) was calculated to determine the combined effect. And the bactericidal effect of fusidic acid combined with ursolic acid was studied by the time-killing curves. The agar double dilution method was used to determine the anti-drug resistance mutation concentration (MPC) and anti-drug resistance mutation selection window (MSW) of fusidic acid alone and in combination with ursolic acid. The viable count of biofilm carrier were determined by serial dilution method and the semi-quantitative biofilm by crystal violet staining method. RESULTS: The combined use of ursolic acid and fusidic acid for SA and MRSA FICI were 0.312 5 and 0.375, respectively. The time-killing curve showed that 1MIC ursolic acid combined with 1MIC fusidic acid has a synergistic bactericidal effect on SA and MRSA. The MPC of fusidic acid to MRSA was 256 μg/mL and the MSW was 256. After fusidic acid combined with ursolic acid, the MPC decreased to 8 μg/mL. The combined group was significantly reduced compared to the fusidic acid group. The semi-quantitative and biofilm bacterial counts of combined group were markedly decreased compared to the fusidic acid group after the biofilm cultivate for 48 h and 72 h.CONCLUSION: The combined use of UA and FA has a synergistic effect on SA and MRSA.

OBJECTIVE: To observe effects of three-dimensional EH composite template on repairing cranial bone defect. METHODS: A total of 53 inpatients were analyzed retrospectively at the Department of Neurosurgery, Taixing People's Hospital from July 2004 to May 2007, comprising 31 males and 22 females, aged 19-68 years. They were in accordance with cranial repairing indication. Reasons for skull defect: traumatic brain injury (41 cases), hypertension (7 cases), intracranial cancer (3 cases), intracranial aneurysm (2 cases). Thirteen patients developed bilateral cranial defect. The defect region: frontal (5 slices, comprising 2 with severe orbital part defect), temporal (9 slices), frontotemporal (42 slices), temporoparietal (3 slices), parietoocipital (4 slices),frontoparietal (3 slices). The smallest single-chip defect size was 4 cm × 6 cm, and the largest one was 12 cm×18 cm. All the patients were mended by three-dimensional EH composite template using spiral CT (≤5 mm thickness) scanning, CAD three dimensional reconstruction, quickly modeling technique. Infection of incisional wound, rejection and complications were observed. RESULTS: A total of 53 patients were followed up, with an average follow-up period of 18 months, and all of them were satisfied with the shapes (100%). Mild collection of the fluid under the scalp was found in 2 cases 4 days and 1 week following surgery, and disappeared after suction and pressure dressing. No other frequent complications were observed after cranioplasty. CONCLUSION: Three-dimensional EH composite template is an ideal material for cranial bone defect, because of its good biocompatibility, easy operation, good postoperative shape and less complications.

OBJECTIVETo explore the relationship of vascular endothelial growth factor C (VEGF-C) and collagen triple helix repeat containing 1 (CTHRC1) expression with the carcinogenesis and prognosis of rectal cancer.

METHODSCancer tissue samples from 120 rectal cancer patients confirmed by pathology in the People's Hospital of Yichun City from September 2005 to September 2010 were included in the study. Expressions of CTHRC1 and VEGF-C were examined by immunohistochemistry and their correlations with clinicopathological features and prognosis were analyzed.

RESULTSThe expression of VEGF-C was positively correlated with tumor size (r=0.943), TNM stages (r=0.784) and tumor differentiation (r=0.773) (all P<0.05). Similarly, the expression of CTRHC1 was also positively correlated with tumor size (r=0.829), TNM stages (r=0.632) and tumor differentiation (r=0.532) (all P<0.05). Rectal cancer patients with low expression of VEGF-C and CTHRC1 had significantly longer survival than those with high expression of VEGF-C and CTHRC1 [(40.0±1.3) vs. (35.4±0.5) months, P<0.01, (39.0±0.5) vs. (35.0±0.5) months, P=0.014].

CONCLUSIONVEGF-C and CTHRC1 may synergistically promote the invasion and metastasis of human rectal cancer, and provide evidence in predicting the prognosis of patients.

Adult ; Aged ; Aged, 80 and over ; Extracellular Matrix Proteins ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Rectal Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; metabolism ; Young Adult

OBJECTIVETo establish an accurate, fast and simple screening method for AZF microdeletions using capillary technology and use it for clinical testing.

METHODSFor each pair of primers, the 5' end of either forward or reverse primer was labeled with a FAM, JOE or TAMRA fluorescence dyes to establish multiplex quantitative fluorescence PCR systems for the establishment of a screening method of Y chromosome AZF microdeletions by capillary technology. The detection of Y chromosome AZF microdeletion was carried out on 725 cases of non-obstructive azoospermia, oligospermia or asthenospermia.

RESULTSA screening method for Y chromosome AZF microdeletions using capillary technology was established. Thirty eight cases of AZF microdeletions were found among 725 cases of non-obstructive azoospermia, oligospermia or asthenospermia, which gave a deletion rate of 5.24%. Y chromosomal microdeletions were found in 8.62% of the azoospermia group, 6.75% of the oligozoospermic group, and 2.23% of the asthenospermia group.

CONCLUSIONAn accurate, fast and simple screening method of Y chromosome AZF microdeletions by capillary technology has been established, which may have an important clinical value.

Adult ; Azoospermia ; genetics ; Capillary Action ; Chromosome Deletion ; Chromosomes, Human, Y ; Humans ; Infertility, Male ; Male ; Multiplex Polymerase Chain Reaction ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; diagnosis

BACKGROUND: Radiation (IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment (TME). Wee1, a cell cycle regulator, can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1 (CDK1). Meanwhile, programed death-1/programed death ligand-1 (PD-1/PDL-1) blockade is closely related to TME. This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody (anti-PD-1 Ab) on radiosensitization of hepatoma. METHODS: The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay on human and mouse hepatoma cells HepG2, Hepa1-6, and H22. The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro . A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice, which were divided into control group, IR group, AZD1775 group, IR + AZD1775 group, IR + anti-PD-1 Ab group, and the IR + AZD1775 + anti-PD-1 Ab group. Cytotoxic CD8 + T cells in TME were analyzed by flow cytometry. RESULTS: Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro . AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage. AZD1775 treatment also reduced the proportion of PD-1 + /CD8 + T cells in the spleen of hepatoma subcutaneous xenograft mice. Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferon γ (IFNγ) + or Ki67 + CD8 T cells and decreasing the levels of CD8 + Tregs cells in the tumor and spleen of the hepatoma mice model, indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγ expression, enhancing CD8 + T cells proliferation, and weakening CD8 + T cells depletion. CONCLUSIONS This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improving cytotoxic CD8 + T cells in TME.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/radiotherapy* ; Cell Cycle Proteins/metabolism* ; Protein-Tyrosine Kinases/genetics* ; Apoptosis ; Programmed Cell Death 1 Receptor ; Cell Line, Tumor ; G2 Phase Cell Cycle Checkpoints ; Liver Neoplasms/radiotherapy* ; Tumor Microenvironment ; Pyrazoles ; Pyrimidinones

Anti-Inflammatory Agents ; therapeutic use ; Antioxidants ; therapeutic use ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Herbal Medicine ; methods ; Humans ; Hypoglycemic Agents ; therapeutic use