OBJECTIVE:To prepare total flavonoids of Hippophae rhamnoides(TFH)-PVP K30 solid dispersion,and to char-acterize and study its in vitro dissolution. METHODS:Solvent method was used to prepare TFH-PVP K30 solid dispersion with dif-ferent drug-loading ratio of 1:1,1:2,1:3,1:4,1:5;single factor test was designed to screen drug-loading ratio using dissolution parameter Td as index;orthogonal test was designed to optimize ultrasonic time,temperature of water bath and drying time for prep-aration technology using in vitro dissolution rate as index,and then validated. SEM,DSC and FT-IR were used to characterize sol-id dispersion. RESULTS:Td of TFH-PVP K30 solid dispersion was the lowest when drug-loading ratio was 1:3. Optimal technolo-gy was ultrasonic time 10 min,temperature of water bath 60 ℃ and drying time 12 h. 90 min accumulative dissolution rate of pre-pared TFH-PVP K30 solid dispersion was 90.22% in average(RSD=1.74%,n=3). The results of SEM,DSC and FT-IR showed that the drug as amorphous form dispersed in the PVP K30,the formation of hydrogen bond of the both. CONCLUSIONS:TFH-PVP K30 solid dispersion is prepared successfully,and in vitro dissolution rate of it is improved significantly.

Objective To evaluate the effect of the bladder scanner upon maintaining the consistency of bladder filling in cervical cancer patients during the intensity-modulated radiotherapy.Methods The bladder volume change of 20 patients diagnosed with cervical cancer throughout radiotherapy were retrospectively analyzed to determine the timing of introducing the bladder scanner. Ten patients undergoing cervical cancer radiotherapy were selected to analyze the consistency between the bladder volume measured by bladder scanner and cone-beam CT (CBCT).The changes of bladder volume before and after the intervention of bladder scanner were statistically compared. Results In total,100 CBCT images of 20 patients were retrospectively analyzed. Nonparametric test demonstrated that the bladder volume significantly differed compared with the planning volume ( P< 0. 05). Bland-Altman plot illustrated high consistency between the bladder volume measured by the bladder scanner and CBCT images with a mean difference of-6. 66 cm3 (95％CI: - 53. 1-39. 83 cm3 ). Paired-t test showed there was statistical difference between the bladder volume before intervention and the planning bladder volume (P= 0. 000).The bladder volume after intervention did not significantly differ from the planning bladder volume (P= 0. 745). Conclusions The bladder volume significantly varies throughout the treatment process. Bladder scanner should be utilized prior to treatment. The bladder volume measured by the portable bladder scanner is consistent with the planning bladder volume.

BACKGROUND: Radiation (IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment (TME). Wee1, a cell cycle regulator, can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1 (CDK1). Meanwhile, programed death-1/programed death ligand-1 (PD-1/PDL-1) blockade is closely related to TME. This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody (anti-PD-1 Ab) on radiosensitization of hepatoma. METHODS: The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay on human and mouse hepatoma cells HepG2, Hepa1-6, and H22. The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro . A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice, which were divided into control group, IR group, AZD1775 group, IR + AZD1775 group, IR + anti-PD-1 Ab group, and the IR + AZD1775 + anti-PD-1 Ab group. Cytotoxic CD8 + T cells in TME were analyzed by flow cytometry. RESULTS: Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro . AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage. AZD1775 treatment also reduced the proportion of PD-1 + /CD8 + T cells in the spleen of hepatoma subcutaneous xenograft mice. Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferon γ (IFNγ) + or Ki67 + CD8 T cells and decreasing the levels of CD8 + Tregs cells in the tumor and spleen of the hepatoma mice model, indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγ expression, enhancing CD8 + T cells proliferation, and weakening CD8 + T cells depletion. CONCLUSIONS This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improving cytotoxic CD8 + T cells in TME.
Humans ; Animals ; Mice ; Carcinoma, Hepatocellular/radiotherapy* ; Cell Cycle Proteins/metabolism* ; Protein-Tyrosine Kinases/genetics* ; Apoptosis ; Programmed Cell Death 1 Receptor ; Cell Line, Tumor ; G2 Phase Cell Cycle Checkpoints ; Liver Neoplasms/radiotherapy* ; Tumor Microenvironment ; Pyrazoles ; Pyrimidinones

OBJECTIVE： To study the improvement effect and mechanism of methylated urolithin A on oleic acid-induced lipid accumulation in human liver cancer Huh-7 cells. METHODS： Oleic acid was adopted to induce lipid accumulation model cells. Huh-7 cells were divided into control group （culture medium）， model group （1 mmol/L oleic acid）， low-dose group （1 mmol/L oleic acid+10 μmol/L methylated urolithin A） and high-dose group （1 mmol/L oleic acid+20 μmol/L methylated urolithin A）. Oil red O staining was used to observe lipid accumulation in cells. Triglyceride（TG） enzyme assay was applied to determine the TG content in cells. PCR was employed to detect the mRNA expression of FASN， SREBP-1， PPAR-α and PPAR-γ in cells. Western blotting was used to determine the protein expression of FASN in cells. RESULTS： After induced by oleic acid， a large amount of lipid droplet accumulated around the cells； the intracellular lipid and TG content， mRNA expression levels of FASN， SREBP-1 and PPAR-γ， protein expression levels of FASN were increased significantly， while mRNA expression level of PPAR-α was decreased significantly （P＜0.01）. After intervened with methylated urolithin A， lipid droplet around the cells decreased significantly； the contents of lipid and TG in cells were decreased significantly， while the mRNA expression levels of FASN， SREBP-1 and PPARγ and protein expression level of FASN were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Methylated urolithin A can improve oleic acid-induced lipid accumulation in Huh-7 cells， the mechanism of which may be associated with inhibiting fat synthesis， promoting lipid metabolism and down-regulating the expression of metabolism-related factors as FASN， SREBP-1 and PPAR-γ.

L-2-aminobutyric acid (L-ABA) is an important chemical raw material and chiral pharmaceutical intermediate. The aim of this study was to develop an efficient method for L-ABA production from L-threonine using a trienzyme cascade route with Threonine deaminase (TD) from Escherichia. coli, Leucine dehydrogenase (LDH) from Bacillus thuringiensis and Formate dehydrogenase (FDH) from Candida boidinii. In order to simplify the production process, the activity ratio of TD, LDH and FDH was 1:1:0.2 after combining different activity ratios in the system in vitro. The above ratio was achieved in the recombinant strain E. coli 3FT+L. Moreover, the transformation conditions were optimized. Finally, we achieved L-ABA production of 68.5 g/L with a conversion rate of 99.0% for 12 h in a 30-L bioreactor by whole-cell catalyst. The environmentally safe and efficient process route represents a promising strategy for large-scale L-ABA production in the future.
Aminobutyrates ; chemical synthesis ; Bacillus thuringiensis ; enzymology ; Candida ; enzymology ; Escherichia coli ; enzymology ; Formate Dehydrogenases ; metabolism ; Leucine Dehydrogenase ; metabolism ; Threonine ; metabolism ; Threonine Dehydratase ; metabolism