MetFab-DOX reduces doxorubicin-resistance of HepG2 / DOX human hepatocellular carcinoma cells / 医学研究生学报

Objective MetFab-DOX can inhibit the proliferation of hepatocellular carcinoma HepG2 cells, but few researches have been conducted on the effect of MetFab-DOX on doxorubicin-resistant HepG2 cells. This study aimed to constructed doxorubicin-resistant HepG2 cell lines and explored the effect of MetFab-DOX on their drug resistance.Methods Using high-dose intermittent induction, we constructed the doxorubicin-resistant hepatocellular carcinoma cell model HepG2/DOX and divided the cells into a blank control, a DOX (5 μg/mL), and an MetFab-DOX group (containing 5 μg/mL doxorubicin). After treatment, we detected the effects of MetFab-DOX on the proliferation, apoptosis, internalization and biological function of the HepG2/DOX cells by CCK8 assay, FCM, cell immunofluorescence, wound healing assay and Transwell invasion assay, respectively. We also established a tumor-bearing model in the nude mouse and examined the effects of MetFab-DOX on the volume and morphology of the tumor.Results The drug resistance index of the HepG2/DOX cells treated with DOX and MetFab-DOX was markedly reduced, with statistically significant difference between the HepG2 and HepG2/DOX cells (P<0.05). After 24 hours of treatment, the cell apoptosis rate was remarkably higher in the MetFab-DOX than in the DOX group (19.87% vs 8.09%, P<0.05), and so was it at 48 hours (41.27% vs 16.15%, P<0.01). The internalization in the cells showed no statistically significant difference between the MetFab-DOX and DOX groups at 30 minutes, while the fluorescence intensity of doxorubicin was markedly higher in the former than in the latter group at 60 and 120 minutes. The cell scratch healing rate was lower in the MetFab-DOX than in the DOX and blank control groups at 24 hours (14.46% vs 16.80% and 19.88%, P<0.05), but higher in the former than in the latter two groups at 48 hours (22.60% vs 36.96% and 56.43%, P<0.01). The number of the membrane-penetrating cells per visual field was significantly decreased in the MetFab-DOX and DOX groups as compared with that in the blank control (646.18 and 880.51 vs 1043.52, P<0.05), and even lower in the MetFab-DOX than in the DOX group (P<0.05). After 40 days of treatment, the tumor inhibition rate was remarkably higher in the MetFab-DOX than in the DOX group (64% vs 35.27%, P<0.05). In the blank control group, the transplanted tumor cells were irregularly arranged and proliferative tumors varied in volume and constituted a larger proportion. The proliferation of the cells was slightly reduced in the DOX group as compared with that in the control. In the MetFab-DOX group, the tumor cells showed a significant shrinkage and a decreased number.Conclusion MetFab-DOX can effectively reduce the doxorubicin-resistance of hepatocellular carcinoma, and the underlying mechanism may be associated with its abilities of increasing the accumulation in drug-induced cells and inducing cell apoptosis.

The action mechanisms of Morus alba leaves extract for the treatment of diabetes based on plasma metabolomics / 药学学报

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.

Effect of different penetration enhancers on percutaneous absorption of active components groups in Shaofu Zhuyu Transdermal Patches / 中草药

Objective: To study the effect of different penetration enhancers on the percutaneous absorption of nine active components in Shaofu Zhuyu Transdermal Patches (SZTP) across skin of mice and optimize the proper penetration enhancers in order to provide the scientific basis for SZTP research, clinical medication, and reform of the traditional forms. Methods: Using improved Franz diffusion cells and taking isolated rat abdomen skins as transdermal barrier, the concentration of these components was determined by UPLC-MS/MS, then the total factor scores of the concentrations at different time were calculated using PCA and employed instead of the concentration to compute the cumulative amounts and steady fluxes. Results: The results showed that compared to the control group, the steady fluxes of the other groups increased significantly, and furthermore, 2.5% azone with 2.5% propylene glycol manifested the best effect. The nine components could penetrate through skin well under the action of penetration enhancers. Conclusion: Azone and propylene glycol could significantly promote the percutaneous penetration effect of the nine active components in SZTP, and it could provide a scientific basis for the preparation research of SZTP.

Fast identification of mycobacteria in microtiter liquid culture / 中华预防医学杂志

<p><b>OBJECTIVE</b>This research was to establish a method for fast identification of mycobacteria in microtiter liquid culture and to evaluate its clinical value.</p><p><b>METHODS</b>2-thiophenecarboxylic acid hydrazide (TCH) and paranitrobenzoic acid (PNB) at different concentrations were added into liquid culture in 96-well plate. Different mycobacterium standard strains were incubated in liquid culture with PNB and TCH for 7 to 10 days. According to the growth assay for 15 mycobacterium strains and 30 mycobacterium tuberculosis strains, the best PNB and TCH concentration were determined. A total of 424 clinical mycobacterium isolates were identified by microtiter liquid culture at the best PNB and TCH concentration. The results of microtiter liquid culture were compared with those of PCR and DNA sequencing.</p><p><b>RESULTS</b>The best concentration of PNB was 200 µg/ml in microtiter liquid culture. Compared with the results of PCR, the sensitivity and specificity for identification of mycobacterium tuberculosis complex in microtiter liquid culture were 97.8% (306/313) and 100.0% (107/107) respectively and those for non-tuberculosis mycobacteria in microtiter liquid culture were 100.0% (107/107) and 96.5% (306/317) respectively. The best concentration of TCH was 0.5 µg/ml. Compared with the results of PCR, the sensitivity of mycobacterium tuberculosis in microtiter liquid culture was 100.0% (305/305). The specificity remained under and more studies were needed.</p><p><b>CONCLUSION</b>In microtiter liquid culture with PNB and TCH, mycobacteria can be identified in 7 to 10 days. The results were accurate and the process was simple without expensive equipments. This method meets clinical needs and can be used in all level hospitals in China.</p>

Evaluation of microscopic observation drug susceptibility for drug susceptibility testing of mycobacterium tuberculosis in smear-positive sputum / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate microscopic observation drug susceptibility (MODS) for mycobacterium tuberculosis drug susceptibility in smear-positive sputum.</p><p><b>METHODS</b>Drug susceptibility of mycobacterium tuberculosis in 275 smear-positive sputum samples collected from TB patients were detected directly by MODS. The susceptibility of seven antimicrobials including streptomycin, isoniazid, rifampicin, ethambutol, levofloxacin, amikacin and capromycin were detected MODS. At the same time the sputum sample were cultured in MGIT 960 tube and the positive isolates were tested for drug susceptibility by MGIT 960 system. The results of MODS were analyzed and compared with that of MGIT 960.</p><p><b>RESULTS</b>Of 275 smear-positive sputum, MODS detected 235 (85.45%). Results of MODS were obtained in a median time of 18 days (5 - 39 d). For the 235 MODS-positive samples, the compliance rates of MODS to MGIT of 7 drugs were 90.21% (212/235), 88.09% (207/235), 93.62% (220/235), 87.23% (205/235), 92.34% (217/235), 88.51% (208/235) and 86.81% (204/235) respectively. The sensitivity of MODS method were 83.33% (90/108), 85.11% (120/141), 90.74% (98/108), 85.71% (78/91), 86.73% (85/98), 76.92% (40/52) and 77.08% (37/48). The specificities of MODS method were 96.06% (122/127), 92.55% (87/94), 96.06% (122/127), 88.19% (127/144), 96.35% (132/137), 91.80% (168/183) and 89.30% (167/187) respectively.</p><p><b>CONCLUSION</b>MODS is an optimal alternative method for direct and rapid drug susceptibility of sputum with high accuracy in a timely and affordable way in resource-limited settings.</p>