Treatment with b-AP15 to Inhibit UCHL5 and USP14 Deubiquitinating Activity and Enhance p27 and Cyclin E1 for Tumors with p53 Deficiency.

Background: TP53 protein is lost or mutated in about half of all types of human cancers and small molecules to regulate mutant p53 repair, or interrupt ubiquitination degradation of p53 induced by E3-ubiquitin ligase Mdm2 have a potential application in clinical application. Methods: To inhibit the deubiquitinase activity of 19S proteasome and restore the p53 protein level, in this study, we utilized p53 knockout mice to test the anti-cancer effect of a specific USP14 and UCH37 inhibitor b-AP15. Results: Our results show that UCHL5, USP14 and COPS5 are upregulated in p53-related tumors, and higher expression of these genes results in a shorter overall survival in patients with p53 deficiency. Treatment with b-AP15, a UCHL5 and USP14 deubiquitinating activity inhibitor in 19S regulatory subunit, induces tumor regression and prolong the survival period of tumor-loaded mice through down-regulation of COPS5 and its downstream AP-1 and E2F1, and up-regulation of the cell cycle-related proteins p27 and Cyclin E1. Conclusions: Thus, our results suggested that inhibition of UCHL5 and USP14 deubiquitinating activity in 19S proteasome may contribute an extensive approach to preventing tumor progress due to p53 deficiency.

Wnt/ß-catenin signaling plays an important role in the protective effects of FDP-Sr against oxidative stress induced apoptosis in MC3T3-E1 cell.

Strontium fructose 1,6-diphosphate (FDP-Sr) is a new strontium-containing compound. The primary aim of this study was to clarify whether the structure component of FDP-Sr, FDP could benefit the protective effect of Sr (II) against oxidative stress induced apoptosis, and meanwhile to further explore the important role of Wnt/ß-catenin signaling in the anti-apoptosis effect of FDP-Sr in response to oxidative stress induced by H2O2 in an osteoblastic MC3T3-E1 cell line. Results showed that FDP-Sr could improve the osteoblastic differentiation under oxidative stress with induced cell proliferation and improved mineralization. The inhibition effect of FDP-Sr on cell apoptosis induced by H2O2 was proved by reduced reactive oxygen species production and activated caspase3. Under oxidative stress, mRNA and protein levels of phospho-ß-catenin reduced, while ß-catenin increased in the FDP-Sr treatment cell, leaded to the up-regulations of Runx2 and OPG at both mRNA and protein levels, finally improved the differentiation of osteoblasts. By the engagement of Wnt/ß-catenin pathway's inhibitor (XAV-939), the protective effects of FDP-Sr on osteoblastic differentiation against oxidative stress were repressed along with inhibited wnt/ß-catenin signaling and reduced mRNA and protein levels of Runx2 and OPG. In conclusion, FDP-Sr was demonstrated to protect osteoblast differentiation from oxidative damage induced by H2O2 through up-regulation of Wnt/ß-catenin signaling, and FDP in FDP-Sr was able to directly improve the oxidative stress injury through its ROS scavenging ability.

[Influence of electroacupuncture intervention on serum ghrelin content and bone mineral density in ovariectomized rats].

OBJECTIVE: To observe the influence of electroacupuncture (EA) on serum ghrelin content and bone mineral density in ovariectomized (OVX) rats so as to explore its mechanism underlying improvement of postmenopausal osteoporosis. METHODS: Thirty ovariectomized SD rats with osteoporosis were randomized into model, EA and estrogen groups (n = 10) and other 10 rats received sham operation were assigned to be control group. For OVX rats of the EA group, EA (2 Hz, 1-3 mA) was applied to "Sanyinjiao" (SP 6) + "Guanyuan" (CV 4) or "Shenshu" (BL 23) + "Housanli" (ST 36), alternately for 20 min, once daily for consecutive 3 months. For rats of the estrogen group, subcutaneous injection of estradiol benzoate (0.1 mg/kg) was given once a week for 3 months. The bone mineral density (BMD) of the lumbar vertebrae and femur were detected by using a bone densitometer, and serum ghrelin content was assayed by ELISA. RESULTS: In comparison with the sham operation group, serum ghrelin level of the model group was significantly increased, and BMD of both lumbar vertebrae and femur were significantly decreased (P < 0.05). Compared with the model group, the serum ghrelin levels were considerably decreased in both EA group and estrogen group, while BMD levels were significantly upregulated in the EA and estrogen groups (P < 0.05). The effects of the EA group were obviously inferior to those of the estrogen group (P < 0.05). CONCLUSION: Acupuncture intervention can improve bone density and lower serum ghrelin level in OVX rats, which may contribute to its effect in improving osteoporosis.

[Clinical observation on acupuncture for treatment of male osteoporosis].

OBJECTIVE: To compare the therapeutic effects of acupuncture combined with drug and simple drug for treatment of male osteoporosis. METHODS: Fifty-five cases were divided into an observation group (25 cases) and a control group (30 cases) randomly. The observation group was treated with acupuncture and moxibustion at Pishu (BL 20), Shenshu (BL 23), Mingmen (GV 4), Shenque (CV 8) and so on combined with taking Alendronate, while the control group was treated with taking Alendronate simply. The improvement of both Integral of Clinical Symptoms (ICS) and Bone Mineral Density (BMD) of two groups was observed after 6 months treatment. RESULTS: The ICS of two groups after treatment both decreased significantly (both P < 0.001), and the decreasing degree in observation group was more significant than that in control group (P < 0.001). The BMD of lumbar vertebrae and femur in observation group increased obviously than that before treatment (P < 0.01, P < 0.05). The increasing degree of BMD of lumbar vertebrae in observation group after treatment was more obvious than that in control group (P < 0.05). There were abdominal pain, diarrhea, nausea, vomiting, dyspepsia and other adverse reaction in control group, while the degree and occurrence rate of those in observation group alleviated and decreased obviously. CONCLUSION: The effect of acupuncture combined with drug for treatment of male osteoporosis is good with little adverse reaction. This method is better than taking Alendronate.