OBJECTIVETo study mRNA expression of receptor activator nuclear factor kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG) in peri-implant tissue during unloading period.

METHODSAn animal model of dental implant was established in 6 male Beagle dogs of 1-2 years old. Bone remodeling was tested at 3, 7, 15, 30, 60 and 90 days since the placement of implants. RANKL and OPG mRNA expression were quantified by real-time polymerase chain reaction (PCR). Then mandibular bones were taken out and the morphological changes were observed by X-ray, bone tissue was tested by immunohistochemistry stain.

RESULTSThe most prominent period of bone remodeling occurred at 7th day after the placement of implants. The expression of RANKL and OPG increased in a time-dependent manner in both soft and hard tissue. After 7 days they gradually decreased.

CONCLUSIONRANKL and OPG can express in soft tissue, and the changing tendency is consistent with the change of bone remodeling, it indicates that RANKL and OPG play an important role in the bone remodeling.

Animals ; Bone Remodeling ; Bone and Bones ; Carrier Proteins ; Dogs ; Male ; NF-kappa B ; Osteoprotegerin ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B

Objective To explore miR-362-3 p expression in laryngeal cancer tissues and its effect on migration of Hep2 cells. Methods miR-362-3 p expression in 50 pairs of tumor and adjacent normal tissues was detected by real-time PCR after sample collection. The relationships between miR-362-3 p expression and clinical pathological characteristics in patients with laryngeal cancer were analyzed.mi R-362-3 p mimic, inhibitor, and control microRNA were transfected into Hep-2 cells. Transfection efficiency was determined by real-time PCR. Wound-healing and transwell assays were used to evaluate Hep-2 migration. Results miR-362-3 p expression was significantly higher in cancer tissues than in adjacent normal tissues (P ＜ 0.05). miR-362-3 p expression was statistically significantly related to node metastasis and clinical stage. Transfection with miR-362-3 p mimic and inhibitor significantly increased and decreased Hep-2 cell migration, respectively (both P ＜ 0.05). Conclusion miR-362-3 p is up-regulated in laryngeal cancer tissues and promotes laryngeal cancer cell migration, suggesting that it acts as a potential oncogene in laryngeal cancer.

Objective To explore the efficacy of the single versus combination drug therapies for benign prostatic hyperplasia(BPH) combined with overactive bladder(OAB).Methods A total of 471 outpatients with BPH and OAB meeting the inclusion/exclusion criteria were enrolled in this prospective cohort study from March 2012 to October 2015.Patients were divided into two groups:(1) the single alpha-blocker treatment group (prostate volume ＜ 30 ml),and (2) the 5 alpha reductase inhibitors(5-ARIs) plus alpha-blocker combination treatment group(prostate volume ≥ 30 ml).The 318 patients were treated with alpha blockers for 4 weeks,and then received a continuing alpha-blocker treatment for 8 weeks if IPSS score changes were less than 30％ (i.e.single alpha-blocker treatment group).And 153 patients were treated with 5-ARIs for 12 weeks,then received 5-ARIs plus alpha-blocker combination treatment for another 4 weeks(a total of 16 weeks)if IPSS score changes were less than 30 ％ (i.e.combination treatment group).The improvements of post-voiding residual(PVR),PV,maximum urinary flow rate(Qmax),international prostate symptom score(IPSS),overactive bladder symptom score (OABSS),quality of life (QOL),urine storage period symptom score (USPSS) and voiding symptom score(VSS)were compared between the two groups.Results The values of IPSS,OABSS,QOL,USPSS and VSS index in the two groups were improved after treatment as compared with pre-treatment(all P≤0.05).Patients in combination treatment group had little improvement in PVR and Qmax after treatment.The OAB symptom remission rates of BPH patients with OAB in single alpha-blocker treatment group were 70.5％ (206/292)and 78.6％ (165/210)after 4 and 12 weeks of treatment respectively.The OAB symptoms remission rates of BPH patients with OAB in combination treatment group were 54.5 ％ (64/122) and 67.1％ (53/79) after 12 and 16 weeks of treatment respectively.Conclusions Both single alpha-blocker treatment and alpha-blocker plus 5ARIs combination treatment,which identification was based on prostate volume,have good effects on BPH patients with OAB.The single alpha-blocker treatment can improve PVR and Qmax,and the alpha-blockers plus 5ARIs combination treatment can improve the prostate volume in BPH patients with OAB.

OBJECTIVE：To establish the method for the con tent determination of related substances in belinstat. METHODS ： HPLC method was adopted and the principal component self-control comparison method with correction factor was used to calculate the contents of related substances. The determination was performed on ODS-AM column with 1.02％ potassium dihydrogen phosphate buffer （pH value adjusted to 3.5 with phosphoric acid ）-acetonitrile（85∶15，V/V）as mobile phase A ，1.02％ potassium dihydrogen phosphate buffer （pH value adjusted to 3.5 with phosphoric acid ）-acetonitrile（30 ∶ 70，V/V）as mobile phase B （gradient elution ），at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and the detection wavelength was 220 nm. The sample size was 10 μL. RESULTS：The linear ranges of belinstat and impurities A ，D，F，G，H were 0.113-1.693， 0.050-1.496，0.117-1.750，0.098-1.471，0.120-1.799，0.100-1.506 μ g/mL（r≥0.999 7）. The correction factors of the last 5 impurities were 1.0，1.0，1.2，1.5，1.0；the detection limits were 0.250，0.590，0.490，0.600，0.500 ng，respectively. The quantification limits were 0.500，1.170，0.980，1.200，1.000 ng，respectively. The recoveries were 90.18％-111.48％（RSD＝ 1.52％-4.78％，n＝9）. RSDs of stability （100 h）and precision tests were no more than 16％，and the durability was good. Impurities A ，D and H were detected in 3 batches of belinlestat ，the contents were 0.030％-0.038％，0.019％-0.022％ and 0.012％-0.013％，respectively. The contents of other maximum monomer impurities were 0.012％-0.013％ and the total impurities were 0.075％-0.084％. Impurities B ，C，F，G were not detected. CONCLUSIONS ：The method for the content determination of related substances in belinstat has been successfully established ，and the method is accurate and specific.

Objective To establish the quality standard of Gansu granules (GGs) and effectively control the quality of GGs. Methods TLC identification method of Gansu granules was improved by using pinocembrin-7-O-β-D-glucoside as the indicator. In HPLC analysis, Agilent ZorbaxSB-C₁₈ (4.6 mm×250 mm, 5 μ m) was used as a chromatographic column, and acetonitrile-0.5% formic acid solution was used as the mobile phase. The detection wavelength of 280 nm was used to analyze the content of pinocembrin-7-O-β-D-glucoside in GGs. Results The separation of characteristic spots was good and clear when the established TLC method was used to identify Gansu granules. Quantitative analysis showed that the concentration of pinocembrin-7-O-β-D-glucoside has a good linear relationship with the peak area in the range of 2.4-240 µg/ml (r= 0.9999). The average sample recovery rate of pinocembrin-7-O-β-D-glucoside is 100.76% with RSD value of 0.66%. The accuracy was good. Conclusion The TLC identification method and HPLC content determination method established in this experiment have strong specificity and good reproducibility, and can be used as an improved standard for the quality control of Gansu granules.

Objective To establish an HPLC method for simultaneous assay of macrocyclic polyphenols from Penthorum chinense Pursh, pinocembrin-7-O-[4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PHG), pinocembrin-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PGHG) and pinocembrin dihydrochalcone-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucoside or thonningianin A (THA), and optimize the extraction process. Methods The total extraction rate of PHG, PGHG, THA was used as an investigated index to analyze the extracts from Penthorum chinense Pursh. Orthogonal design was applied to evaluate solvent amount, extraction time, solvent concentration and extraction times as the influencing factors for the optimal extraction process of macrocyclic polyphenols from Penthorum chinense Pursh. Results When this content assay method was adopted, there were good linear relationships for PHG, PGHG, THA in the linear range. The recoveries were between 100.90% to 102.04% with the RSDs below 1.5%. The optimal extraction process was involved in cutting Penthorum chinense Pursh into 3-5 cm, adding 10 times 80% ethanol aqueous solution by volume and refluxing 2 hours twice. The extraction rate of macrocyclic polyphenols was above 90% with this process. Conclusion This assay method is accurate, stable, and repeatable. The optimized extraction process is stable and feasible for further development and utilization.