BACKGROUND:Currently, bone graft is mainly used for repair of bone defects, and tricalcium phosphate is the most used artificial bone material. But the effectiveness of the tricalcium phosphate bone graft is stil controversial, and there is also no detailed report about its function during the healing of bone defect. ＜br> OBJECTIVE:To observe the concentration changes of bone morphogenetic protein-2 and vascular endothelial growth factor as wel as bone healing after tricalcium phosphate graft in bone defects. ＜br> METHODS:Forty-eight C57 mice were randomly divided to experimental group and control group. A 2-mm-long diaphyseal segment and periosteum from the middle of the right femur was cut to prepare unilateral bone defect models. Tricalcium phosphate bone graft was used in the experimental group, and no bone graft in the control group. During the fol owing 4 weeks, X-ray examination was done once a week to observe the bone healing, and then the animals were executed for col ecting samples in the graft area. The concentrations of bone morphogenetic protein-2 and vascular endothelial growth factor in samples which were taken from the bone graft area were determined by using ELISA assay. ＜br> RESULTS AND CONCLUSION:X-ray films showed that 2 weeks later, bone fracture healed mostly in the experimental group except a smal part of cortical bone;3 weeks later, bone fracture was basical y healed, and only a smal amount of tricalcium phosphate remained;4 weeks later, bone fracture was completely healed, and the cal us grew obviously, and the tricalcium phosphate was nearly absorbed. In the control group, the fracture line was stil visible at 1-2 weeks, but it became vague at 3 weeks;then, the fracture was healed at 4 weeks except some of the cortical bone. The levels of bone morphogenetic protein-2 and vascular endothelial growth factor were significantly higher in the experimental group than in the control group at different time points (P＜0.05). These results suggest that tricalcium phosphate bone graft can up-regulate the expression of bone morphogenetic protein-2 and vascular endothelial growth factor and accelerate bone healing.

Objective To investigate the clinical and genetic characteristics in two patients with androgen insensitivity syndrome. Methods Clinical features and laboratory data were collected from the patients and their families. All exons of the androgen receptor gene were amplified by PCR and PCR products were sequenced. Results Patient 1 presented with unambiguous female external genitalia, unilateral gynecomastia and primary amenorrhea. He did not have axillary hairs or pubic hairs. Patient 2 presented with undervirilization including scanty body hairs, gynecomastia and hypospadias. A missense mutation of

Objective To explore the antibacterial effect and mechanisms of platelets (PLT)on Staphylococcus epidermidis (SE).Methods Built infectious model of SE in vitro,whose final concentration was 105 CFU/ml in the reaction.Separately cocultured these models with PLT (the final concentration 400 × 109/L)-plasma (positive control)and BHI medium 12 hours later,and detected the antibacterial effect of PLT by making a liquid thinner and spreading counting method,and draw-ing antibacterial curve.Meanwhile,observed the bacterial structure by transmission electron microscope (TEM),and initially explored the antibacterial mechanism of PLT.Results The study showed the antibacterial effect of PLT on SE was very ob-vious,which appeared later than the plasma (M)group,but enduring.The images of TEM showed an electronlight region appeared in the centre of bacterium,contained condensed DNA molecules and leaded to slower fission.Conclusion PLT can damage the DNA structure of SE,and then affect the fission of SE,finally inhibit the proliferation of SE.

Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.

Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.

Objective To evaluate the relationship between CD4+ T lymphocyte count and results of enzyme-linked immunospot (ELISPOT) assay in human immunodeficiency virus (HIV)-Mycobacterium tuberculosis (M.tb) coinfected patients.Methods A total of 193 HIV-infected individuals in Yunnan Province and Shanghai were enrolled.T-SPOT.TB assay was employed to detect M.tb specific T lymphocyte in the peripheral blood mononuclear cells (PBMC).CD4+ T lymphocyte in PBMC from the enrolled subjects was detected by flow cytometry.Data were analyzed using t test.ResultsThe incidence of latent tuberculosis in HIV-infected individuals was 30.6％.The CD4+ T lymphocyte counts in HIV-infected individuals with active tuberculosis were 190×106/L,which were significantly lower than those in HIV-infected individuals with latent tuberculosis (484×106/L; t=6.665,P＜0.01).The HIV-infected individuals were stratified according to CD4+ T lymphocyte counts of ＞500×106/L,200×106-500×106/L,and ＜200×106/L and the constituent ratios of active tuberculosis/latent tuberculosis were 1∶16.2,1∶1.3 and 5.6∶1,respectively.Among 79 subjects with positive T-SPOT.TB results,20 were coinfected with active tuberculosis,in which 14 had CD4+ T lymphocyte counts of ＜200 ×106/L,5 had 200×105-500×106/L and 1 had ＞500×106/L.Fifty-two in 59 HIV/latent tuberculosis patients individuals had CD4+ T lymphocyte counts of ＞200×106/L.ConclusionsThe prevalence of latent tuberculosis in HIV-infected individuals is high in China.Cellular immunity in HIV-infected individuals with active tuberculosis is severely impaired.With the decrease of CD4 ′ T lymphocyte counts,patients with latent tuberculosis are prone to develop active tuberculosis in HIV-infected individuals.The negative predictive value of T-SPOT.TB is significantly diminished in patient with low CD4+ T lymphocyte counts,especially less than 200×106/L.

Objective To elucidate the effects of SP600125 at different concentrations on the proliferation and osteo-differentiation of human adipose-derived stem cells (hASCs).Methods The hASCs harvested were cocuhured with SP600125 at concentrations of 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L in growth medium (OM group) and in osteogenesis medium (OM group),respectively.The DNA quantitative assay was carried out to evaluate proliferation of the hASCs;flow cytometry was used to determine the effect of SP600125 on the cell cycles of hASCs;Alkaline phosphatase level (ALP) and calcium deposition tests were conducted to observe the effects of SP600125 at different concentrations on osteogenic differentiation of the hASCs.Results The proliferation of hASCs was inhibited by 42.1％ when the cells were cocultured with SP600125 at the concentration of 10 μmol/L;the suppression decreased with decreased concentration of SP600125.The hASCs of phase G0/G1 in GM cocultured with SP600125 at the concentration of 10 μmol/L were more than those in GM cocultured with dimethylsulfoxide at the same concentration.ALP test revealed that after 10 days of culture in vitro the staining was more and more weakened and scattered and the ALP activity was more and more decreased with the increased concentration of SP600125.The extracellular calcium deposition of hASCs after 14 days of culture in vitro showed that the size and number of calcium nodules decreased with the increased concentration of SP600125.Conclusion SP600125 can suppress the proliferation and osteogenic differentiation of hASCs in vitro.

Dysglycemia is independently associated with the mortality of critically ill patients. Therefore, the management of blood glucose plays an important role in comprehensive therapy. It is suggested that the same target value of blood glucose (7.8-10.0 mmol/L) should not be set for all critically ill patients. Instead, it should be individually set based on the causes of the patient's admission and the status of blood glucose before admission. For this reason, there is an urgent need for a convenient protocol and method to regulate the dosage of insulin. The first hospital of Jiaxing, collaborating with information engineers, developed a modified eProtocol-insulin for domestic population with mathematical modeling and developed an Application Software (APP), which is convenient for clinical use. This is the first eProtocol-insulin and smart device APP for critically ill patients in China.

OBJECTIVETo construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.

METHODSLentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.

RESULTSThe recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).

CONCLUSIONVHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Apoptosis ; Carcinoma, Renal Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

OBJECTIVETo explore the influence of recombinant human growth hormones (rhGH) postburn systemic metabolism.

METHODSTwenty-four burn patients were randomly and equally divided into treatment and control groups. Same amount of rhGH (9 U/d) or isotonic saline was injected subcutaneously to respective patients during 3 approximately 17 postburn days (PBDs). Blood samples were harvested at 3, 10 and 17 PBDs for the determination of serum growth hormone, insulin-like growth factor (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), serum proteins, plasma insulin, plasma glucagons and blood glucose, which were then compared and analyzed between two the groups.

RESULTSThe serum levels of GH, IGF-1, IGFBP-3, serum prealbumin and transferrin in rhGH treatment group were evidently higher than those in control groups at 10 and 17 PBDs (P < 0.05 approximately 0.01). But there was no obvious difference in serum albumin, plasma insulin, glucagon and blood glucose (P > 0.05).

CONCLUSIONSmall dose of rhGH could promote systemic protein synthesis with no side effects on blood glucose levels.

Adolescent ; Adult ; Blood Proteins ; drug effects ; metabolism ; Burns ; blood ; Female ; Growth Hormone ; blood ; pharmacology ; Humans ; Insulin ; blood ; Insulin-Like Growth Factor I ; analysis ; drug effects ; Male ; Middle Aged ; Recombinant Proteins ; pharmacology