The Wnt pathway plays a crucial role in skeletal development and is indispensable for determination of OS cell lines. In recent years, experimental evidences on Wnt signaling pathway in OS cell lines and OS animal models, and that of Wnt signaling pathway as a diagnosis and prognosis marker of OS suggested that Wnt signaling pathway plays an important role in the progression, invasion and metastasis of OS. In addition, the strategy and safety evaluation to target Wnt to treat OS are underway. Exploring the significant role of Wnt signaling pathway in OS may aid in personalizing therapeutics to increase patient survival.

Objective To investigate the relationship between edema and aquaporin 4 (AQP4) within traumatic penumbra (TP) of rats with brain trauma.Methods Eighty-eight healthy adult Wistar rats were randomly divided into control group (n =11) and trauma group (n =77),according to the random number table.Trauma group were further subdivided into seven time points (1 h,6 h,12 h,24 h,48 h,72 h and 7 d) of 11 animals each.Brain tissue samples from the moderate brain models were collected to evaluate brain edema with histological observation,blood brain barrier (BBB) permeability with semiquantitative immunohistohemical staining of IgG,and AQP4 expression with immunofluorescence and Western blotting.Results In control group pathology and IgG staining revealed no abnormalities and expression of AQP4 was few.In trauma group light edema zone was visualized at 1 h,began widening,reached the peak at 12 h [(1.589 ±0.020)mm],and then began narrowing.There were significances in width of the edematous band at each time point except for the comparison at 24 h vs.48 h and 72 h vs.7 d (P ＜ 0.05).After trauma,vasogenic edema was found in edema zone at 1 h,intracellular edema was found at 6 h,both aggravated at 12 h and alleviated slightly at 24 h,and intracellular edema predominated at 48 h.IgG showed intensively positive staining at 1,12 and 48 h,and weak staining at 6 h,24 h,72 h and 7 d.After trauma,expression of AQP4 decreased at 1 h (0.659 ± 0.021),returned slightly at 6 h (1.257 ±0.058),peaked at 12 h (2.499 ±0.136),declined again at 24 h (2.267 ± 0.068),re-raised at 72 h (2.078 ± 0.065),and returned to the baseline at 7 d (1.280 ± 0.065).There were significant differences in level of AQP4 at each time point except for the comparison at6h vs.7 d,24 h vs.72 h and 24 h vs.72 h (P＜0.05).Conclusions In the early phase vasogenic edema characterized by BBB damage is significant within TP,which leads to decreased expression of AQP4.However,the subsequent up-regulation of AQP4 results in intracellular edema,which accelerate the spreading of TP.AQP4 may involve in body's defense reaction.

OBJECTIVE:To prepare a nickel-titanium alloy stent coated by anti-tuberculotic isoniazid and study the dissolution of isoniazid in vitro.METHODS:The polyurethane which was used as film-former was mixed with isoniazid before being coated uniformly on the stent,then the stend was torrefied and dried so that the drug layer alloy stent was obtained.The content and accumulated dissolution in vitro of isoniazid from the stent was determined by ultraviolet spectrophotometry.RESULTS:The linear range of isoniazid was 5.0～30.0 ?g?mL-1(r=0.999 9)and its recovery rate was 99.67%(RSD=1.06%).A high accumluated dissolution rate of isoniazid was achieved,over 60% within the first 30 minutes,followed by a sustained dissolution in the following 8 h.CONCLUSION:The preparation method of the stent is simple and the quality is controllable.

In order to study the effect of nitric oxide (NO) on the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L-arginine (L-Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF-1alpha mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT-PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6 +/- 2.7 mmHg, 1 mmHg=0.133 kPa) was significantly lower than that in chronic hypoxic group (35.8 +/- 6.1 mmHg, t=0.2918, P<0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L-arginine group (24.4 +/- 3.8 mmHg, t=0.2563, P<0.05). ISH showed that the expression of HIF-1alpha mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076 +/- 0.0205) was markedly weaker than that in chronic hypoxic group (0.3317 +/- 0.0683, t=3.125, P<0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L-arginine group (0.1928 +/- 0.0381, t=2.844, P<0.05). RT-PCR showed that the content of HIF-1alpha mRNA in chronic hypoxic group (2.5395 +/- 0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781 +/- 0.3628) and hypoxia plus L-arginine group (1.4511 +/- 0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF-1alpha mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.
Anoxia/metabolism ; Arginine/pharmacology ; Hypertension, Pulmonary/*metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; Nitric Oxide/*pharmacology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Random Allocation ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors/*biosynthesis ; Transcription Factors/genetics

SUMOylation is an important post-translational modification of proteins. Similar to ubiquitylation, SUMOylation is the process that the small ubiquitin-like modifier (SUMO) proteins are specifically and covalently binding to lysine residues of substrate proteins. Through SUMOylation, the physiological functions and pathological processes of cells are well controlled and balanced, and its abnormal activation has been reported in various tumors. Therefore, SUMOylation has been a potential target for anti-tumor drug development. In this review, we summarize recent advances on development of inhibitors targeting SUMOylation pathway and their antitumor properties.

AIM:To observe the changes of nuclear factor-?B (NF-?B) activity and inducible nitric oxide synthase (iNOS) expression in hypoxic pulmonary hypertension(HPH). METHODS:The rat model of HPH was used. The NF-?B activity and iNOS expression in lung tissue were determined by immunohistochemistry(IHC), in situ hybridization (ISH), RT-PCR and Western blot.RESULTS:ISH showed that iNOS mRNA expression in intraacinar pulmonary arteriole (IAPA) in H28 d group(hypoxic treatment for 28 days) was stronger than that in normal group, H5d group and H14 d group. RT-PCR showed that the iNOS mRNA in H28 group was 2.1 times, 1.9 times and 1.8 times higher than that in normal group, H5d group and H14 d group, respectively. The nucleic anti-NF-?B stain was observed in H28 d group, which was significantly stronger, but the I-?B amount was 2.8 times, 2.7 times and 2.5 times lower than that in normal group, H5d group and H14 d group, respectively. CONCLUSION: The activity of NF-?B was correlated with the hypoxic pulmonary vessel structural remodeling and iNOS expression.

AIM: To investigate the expression of phosphatidylinositol 3-kinase (PI-3K) during the proliferation of the hypoxic pulmonary arterial smooth muscle cells(PASMC). METHODS: The cultured PASMC was divided into two groups: serum-free medium (SFM) group and 48 hours hypoxia (H48h) group. Immunohistochmistry(IHC) and Flowcytometer(FCM) were used to detect the cultured PASMC. RESULTS: The positive expression of PI-3K p110 existed in both cultured PASMC groups. The staining intensity in H48h group (0 1891?0 0301) was significantly stronger than that in SFM group (0 1025?0 0164, P

Objective To observe the protein expression related to cognitive and learning memory function, and to investigate the effect of aquaporin 4 (AQP4) silence on learning and memory function in traumatic brain injury (TBI) rats. Methods Ninety-six healthy adult male Wistar rats were divided into groups according to the random number table. ① Forty-eight rats were divided into sham operation (sham) group, TBI group (by using modified Feeney method), AQP4 RNA interference (RNAi) negative group [TBI+meaningless small interfering RNA (siRNA)-AQP4 liposome solution 10 μL], and AQP4 RNAi group (TBI+siRNA-AQP4 liposome solution 10 μL). In each group, brain tissues of 4 rats were harvested at 1, 6 and 12 hours respectively. The protein expressions of hippocampus AQP4, general control nonderepressible 2 kinase (GCN2), cyclic adenosine monophosphate response element binding protein (CREB) and phosphorylated CREB (p-CREB) were detected by Western Blot. ② In addition, 48 rats were divided into normal control group (control group), sham group, TBI group and AQP4 RNAi group, brain water content were measured in 6 of them after 12 hours of injury, and 6 were used in Morris water maze test. Results ① The protein expressions of hippocampus AQP4 and GCN2 in TBI group were significantly higher than those in sham group, and increased gradually with time with statistical difference at 12 hours (AQP4 protein: 5.03±0.09 vs. 1,GCN2 protein: 4.01±0.13 vs. 1, both 1 < 0.01);the protein expressions of hippocampus CREB and p-CREB were significantly lower than those in sham group, and decreased gradually with time with statistical difference at 12 hours (CREB protein: 0.38±0.03 vs. 1, p-CREB protein:0.38±0.03 vs. 1, both 1 < 0.01). Compared with TBI group, the protein expressions of AQP4 in AQP4 RNAi group was significantly decreased (1 hour: 1.02±0.04 vs. 2.23±0.05, 6 hours: 1.23±0.03 vs. 2.59±0.04, 12 hours: 2.20±0.08 vs. 5.03±0.09, all 1 < 0.01), but there were no significant difference in the expressions of GCN2, CREB or p-CREB. There was no significant difference in the expression of protein between AQP4 RNAi negative group and TBI group.② The brain water content in TBI group was significantly higher than that in control group and sham group [(83.7±0.4)% vs. (76.2±0.2)%, (76.2±0.3)%, both 1 < 0.01]. The brain water content in AQP4 RNAi group [(78.8±0.3)%] was significantly decreased as compared with that in TBI group (1 < 0.01). The latency of Morris water maze test was significantly prolonged in the day 11, 13 and 15 after the injury of the TBI group and AQP4 RNAi group, and the exploration time was significantly shortened. Compared with TBI group, the incubation period of AQP4 RNAi group was significantly shortened at 15 days (s: 60.2±11.1 vs. 62.0±11.5, 1 < 0.05), and the exploration time was significantly prolonged (s: 37.0±8.5 vs. 32.7±9.2, 1 < 0.05). Conclusions The impairment of cognitive and learning memory function in rats after TBI was significantly related to the changes in CREB and GCN2 in cognitive and learning memory function. After RNAi treatment, the cognitive and learning and memory function of rats was not improved obviously, but the brain edema could be alleviated.

The ultrasonographic appearances of osteosarcomas and the roles of ultrasonography in the diagnosis and surgical staging of osteosarcomas were investigated. A comparative study was performed on 45 cases of osteosarcomas by ultrasonography and radiography. Bony changes, periosteal reaction and soft tissue mass were evaluated for each lesion. The results showed that ultrasonography revealed a solid mass around bone in 42 patients, bone destruction in 24 patients and periosteal reaction in 16 patients. Plain radiographs showed bony changes in 44 patients and no bony change in remaining one patient, shadowing of soft tissue swelling in 30 patients, and pulmonary metastases in 3 patients. Surgical biopsy and pathological examination confirmed osteosarcoma in all 45 patients. Soft tissue mass was confirmed in 42 patients surgically. The diagnostic accuracy of soft tissue masses by ultrasonography and radiography was 100 % (42/42) and 71.4 % (30/42), respectively. The positive rate of ultrasonography and radiography in displaying bony changes was 53.3 % (24/45) and 97.8 % (44/45), respectively. In conclusion, in the detection of soft tissue mass of osteosarcoma, ultrasonography is superior to radiography, and in displaying bony changes of osteosarcomas, radiography is superior to ultrasonography. So it may come to a conclusion that plain radiography combined with ultrasonography can completely display the bony and soft tissue lesion of osteosarcomas.

Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P<0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P>0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P<0.01). The expression of RANKL was inhibited (P<0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P>0.05), but the expression of RANKL was higher in PMMA group than in control group (P<0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P<0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P<0.01) and the expression of OPG mRNA was significantly increased (P<0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P<0.05), but the expression of OPG mRNA had no significant difference between them (P>0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.