Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.

Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.

OBJECTIVE:To establish a method for the contents determination of tanshinol,protocatechuic aldehyde,ferulic ac-id and salvianolic acid B in Yixinshu capsule. METHODS:Dual-wavelength HPLC was performed on the column of Eclipse XDB-C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280 nm(tanshinol,protocatechuic aldehyde,salvianolic acid B)and 320 nm(ferulic acid),column tempera-ture was 30℃and volume was 10 μl. RESULTS:The linear range of tanshinol,protocatechuic aldehyde,ferulic acid and salvianolic acid B were respectively 9-144μg/ml(r=0.999 9),0.5-8μg/ml(r=0.999 9),0.65-10.4μg/ml(r=0.999 9)and 221.25-3 540μg/ml (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.90%;the average recovery was respective-ly 100.8%(RSD=1.65%,n=9),100.1%(RSD=2.87%,n=9),100.1%(RSD=3.01%,n=9) and 99.4%(RSD=2.05%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Yixinshu capsule.

Objective To investigate effect of the mild CYP3A4 inhibitor aprepitant on the pharmacokinetics of orally administered controlled-release ( CR) oxycodone.Methods This study designed was an single-sequence with two phases in cancer patients with pain who continued to be administered orally with multiple doses of CR oxycodone every 8 or 12 hours.Plasma concentration of oxycodone and its metabolites were measured up to 8 hours after administration as follows: on day 1, CR oxycodone was administered alone; on day 2, CR oxycodone was administered with aprepitant (125 mg, at the same time of oxycodone dosing in the morning).The steady-state trough concentrations ( Css) were measured from day 1 to day 3.Results Aprepitant increased the area under the plasma concentration-time curve (AUC0-8) of oxycodone by 25%(P<0.001) and of oxymorphone by 34% (P<0.001), as well as decreased the AUC0-8 of noroxycodone by 14% (P<0.001).Moreover, aprepitant increased Css of oxycodone by 57%(P=0.001) and of oxymorphone by 36% (P <0.001) and decreased Css of noroxycodone by 24% (P =0.02) at day 3 compared to day 1.Conclusion The clinical use of aprepitant in patients receiving multiple doses of CR oxycodone for cancer pain significantly altered plasma concentration levels, but would not appear to need modification of the CR oxycodone dose.

OBJECTIVE:To establish a method for the content determination of ephedrine hydrochloride in Luofu mountain rheumatism plaster. METHODS:HPLC was performed on the column of Eclipse XDB-C18 with the mobile phase of acetoni-trile-0.1%phosphoric acid(9∶91,V/V),at the flow rate of 1.0 ml/min. The detection wavelength was 207 nm,the column tempera-ture was 40 ℃,and the volume was 5 μl. RESULTS:The linear range of ephedrine hydrochloride was 81.6-408 ng(r=0.999 7);RSDs of precision,stability and repeatability tests were lower than 2%;and the average recovery rate was 96.73%(RSD=1.9%, n=6). CONCLUSIONS:The method is fast and accurate,and can be used for the content determination of ephedrine hydrochlo-ride in Luofu mountain rheumatism plaster.

OBJECTIVE:To establish a method for simultaneous determination of tanshinol,protocatechuic aldehyde,paeoni-florin,ferulic acid and salvianolic acid B in Shenshao oral liquid. METHODS:RP-HPLC was performed on the column of Eclipse XDB C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280,230 and 320 nm,column temperature was 30 ℃ and volume was 10 μl. RESULTS:Under the chro-matographic conditions,5 kinds of components could be completely separated,the linear range of tanshinol,protocatechuic alde-hyde,paeoniflorin,ferulic acid and salvianolic acid B were respectively 24-384 μg/ml(r=0.999 9),1.25-20 μg/ml(r=0.999 9), 40.5-648 μg/ml(r=0.999 8),1.5-24 μg/ml(r=0.999 9),145-2 320 μg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were no more than 2.2%;the average recovery was respectively 100.7%(RSD=1.23%,n=9),100.0%(RSD=2.19%,n=9),99.6%(RSD=0.87%,n=9),100.3%(RSD=1.11%,n=9) and 99.3%(RSD=2.46%,n=9). CONCLUSIONS:The method is specific with good precision and reproducibility,and can be used for the content determination of 5 components in Shenshao oral liquid.

The successful cryopreservation of reproductive cells has important practical significance in many fields. In order to improve the recovery rate and viability of cryopreserved cells, it is necessary to study the permeability characteristics of cell membrane to both water and cryoprotectant. In this paper we review the studies on membrane permeability of animal reproductive cell for the recent years. We firstly list the typical permeability data of spermatozoa and oocyte membrane for water and cryoprotectant. We then analyze the effects of these characteristics on the design of cryopreservation protocol. We also introduce the latest experimental methods to measure the cell membrane permeability.
Animals ; Cell Membrane Permeability ; physiology ; Cryopreservation ; methods ; Female ; Humans ; Male ; Oocytes ; cytology ; Spermatozoa ; cytology

Cryopreservation is essential for the long-term storage and banking of cartilage grafts. This paper reviews the developments on the cryopreservation of cartilage and transplantation of cryopreserved cartilage grafts during the past 10 years. It is stated that the current technologies for cryopreservation of cartilage grafts are not mature. Further systematic studies are necessary.
Animals ; Cartilage ; transplantation ; Cryopreservation ; Humans ; Tissue Preservation ; methods