AIM: To investigate the effect of 3-methyladenine (3-MA) combined with trichostatin A (TSA) on triple-negative breast cancer cells and the mechanism involved.METHODS: The viability of MDA-MB-231 cells was detected by MTT assay, the migration ability was determined by scratch assay, and the expression of autophagy-related proteins was detected by Western blot.RESULTS: TSA significantly inhibited the viability of MDA-MB-231 cells in a time-and dose-dependent manner.The results of scratch assay showed that TSA inhibited cell migration ability.Western blot data indicated that TSA resulted in a moderate increase in LC3-Ⅱ expression.Moreover, 3-MA inhibited cell autophagy induced by TSA, and combination of 3-MA and TSA further inhibited the viability of MDA-MB-231 cells.CONCLUSION: Combination of 3-MA and TSA may effectively inhibit the growth of triple-negative breast cancer cells.

The stable and controllable quality of decoction pieces is an important factor to ensure the efficacy of clinical medicine. Considering the dilemma that the existing standardization of processing mode cannot effectively eliminate the variability of quality raw ingredients, and ensure the stability between different batches, we first propose a new strategy for Chinese medicine processing technologies that coupled with individuation processed and cybernetics. In order to explain this thinking, an individual study case about different grades aconite is provided. We hope this strategy could better serve for clinical medicine, and promote the inheritance and innovation of Chinese medicine processing skills and theories.
Chemistry, Pharmaceutical ; methods ; standards ; Cybernetics ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; toxicity ; Humans ; Quality Control

OBJECTIVETo prepare andrographolide composite particles, and evaluate their particle structure and dissolution.

METHODThe mechanical crushing method was adopted to prepare andrographolide and polyethylene glycol (PEG) 6000 composite particles. The structures were characterized by the scanning electron microscope (SEM) and the differential scanning calorimeter (DSC). The contact angles were determined by the contact angle analyzer. The in vitro dissolution curve was detected.

RESULTAndrographolide and PEG 6000 gave rise to coated composite particle structures, with the decrease in the crystallinity of andrographolide. The in vitro dissolution rate of composite particles was significantly obvious than that of its raw materials, ultrafine powder and their physical mixtures.

CONCLUSIONAndrographolide composite particles based on the mechanical crushing method could notably enhance the in vitro dissolution of andrographolide.

Calorimetry, Differential Scanning ; Chemistry, Pharmaceutical ; methods ; Diterpenes ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Particle Size ; Solubility ; Torsion, Mechanical

In this paper, Rhodiolae Crenulatae Radix et Rhizoma extract,with high hygroscopic,was selected as research model, while lactose was selected as modifiers to study the effect of the grinding modification method on the hygroscopic. Subsequently, particle size distribution, scannin electron microscopy, infrared spectroscopy and surface properties were adopted for a phase analysis. The results showed that the modified extract, prepared by Rhodiolae Crenulatae Radix et Rhizoma extract grinding 5 min with the same amount of lactose UP2, which hygroscopic initial velocity, acceleration, and critical relative humidity moisture were less than that of Rhodiolae Crenulatae Radix et Rhizoma extract and the mixture dramatically. In addition, compared with the mixture, the size distribution of modified extract was much less, the microstructure was also difference, while the infrared spectroscopy and surface properties were similar with that of lactose. It is the main principle that lactose particle adhered to the surface of Rhodiolae Crenulatae Radix et Rhizoma extract after grinding mofication to decress the moisture obviously.
Drugs, Chinese Herbal ; chemistry ; Lactose ; chemistry ; Particle Size ; Rhizome ; chemistry ; Rhodiola ; chemistry ; Spectrophotometry, Infrared ; Surface Properties ; Wettability

BACKGROUNDTo construct recombinant retroviral vector expressing HBcAg in eukaryotic cells.

METHODSThe HBV core gene fragment was amplified by using PCR from pADR which contains complement nucleotide sequence of HBV subtype adr and cloned into retroviral expression plasmid pLXSN, then transfected into packing cell (PT67) with lipofec AMINE. After 2-3 weeks selection with G418, large colonies were isolated and transferred to individual plates. Virus-containing medium was collected and used to infect NIH3T3, EL4 and mouse bone marrow derived dendritic cells(DC). DNA was extracted from infected cells and tested by PCR. Indirect immunofluorescence and FACS were used to detect the expression of HBcAg. Cell mediated immunity of immunized C57BL/6 mice with transduced DC was analyzed.

RESULTSThe insertion of HBV core gene fragment in the recombinant retroviral plasmid was confirmed by PCR as well as enzyme digestion with EcoR1 and BamH2. The viral titer reached 3 x 10(5) CFU/ml. The result of PCR showed that the HBV core gene had been integrated into the genome of infected NIH3T3 cells. Indirect immunofluorescence and FACS analysis showed that HBcAg had been expressed in these cells. HBcAg specific CTL responses could be generated in mice immunized with retrovirus transduced DC.

CONCLUSIONSHBV core gene had been integrated into eukaryotic cells with retroviral vector and target gene had been expressed efficiently. These results may have some impact on gene therapy of chronic hepatitis B.

3T3 Cells ; Animals ; Cloning, Molecular ; Dendritic Cells ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Mice ; Recombination, Genetic ; genetics ; Retroviridae ; genetics ; Transfection

OBJECTIVETo investigate the distribution of tetracycline-arginine-glycine-aspartate-tyrosine (T-RGDY) in mice and its effect on bone.

METHODS125-labeled T-RGDY was studied for its distribution in mice and for its effects on bone by histomorphometry in ovariectomized rats.

RESULTSThe 125I-labeled T-RGDY was more concentrated in the osteoporotic bone than in the normal bone. Compared with ovariectomy group, the morphologic index such as trabecular bone volume/total tissue volume (TBV/TTV), TBV/sponge bone volume (SBV), and mean trabecular plate thickness (MTPT) in T-RGDY group significantly increased (P < 0.05). As compared with sham operation group, MTPT significantly increased in T-RGDY group (P < 0.05), while TBV/SBV and mean trabecular plate density significantly decreased (P < 0.05), and TBV/TYV and mean trabecular plate spacing were almost the same as those in sham operation group (P > 0.05).

CONCLUSIONT-RGDY may concentrate in bone tissue to a certain degree, which is closely related with the status of bone remodeling. T-RGDY may inhibit the bone loss caused by ovariectomy.

Animals ; Bone Density ; drug effects ; Bone Remodeling ; drug effects ; Female ; Mice ; Oligopeptides ; pharmacokinetics ; pharmacology ; Osteoporosis ; metabolism ; prevention & control ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Tetracycline ; pharmacokinetics ; pharmacology ; Tissue Distribution ; Tyrosine ; pharmacokinetics ; pharmacology

Nucleoside phosphorylases (NPases) were found to be induced in Enterobacter aerogenes DGO-04, and cytidine and cytidine 5'-monophosphate (CMP) were the best inducers. Five mmol/L to fifteen mmol/L cytidine or CMP could distinctly increase the activities of purine nucleoside phosphorylase (PNPase), uridine phosphorylase (UPase) and thymidine phosphorylase (TPase) when they were added into medium from 0 to 8 h. In the process of enzymatic synthesis of adenine arabinoside from adenine and uracil arabinoside with wet cells of Enterobacter aerogenes DGO-04 induced by cytidine or CMP, the reaction time could be shortened from 36 to 6 h. After enzymatic reaction the activity of NPase in the cells induced remained higher than that in the cells uninduced.
Cytidine ; pharmacology ; Cytidine Monophosphate ; pharmacology ; Enterobacter aerogenes ; enzymology ; Enzyme Induction ; Pentosyltransferases ; biosynthesis ; Vidarabine ; biosynthesis

Objective:To explore the incidence and risk factors of perioperative deep vein thrombosis (DVT) in patients with intertrochanteric femur fractures.Methods:A retrospective case-control study was used to analyze the clinical data of 485 patients with intertrochanteric femur fracture treated from June 2014 to September 2018 in Honghui Hospital affiliated to Xi'an Jiaotong University Medical College. There were 196 males and 289 females, aged 16 to 102 years, with an average of 74.6 years. All patients were divided into preoperative DVT group, non-DVT group, and postoperative DVT group and non-DVT group according to the results of preoperative and postoperative lower limb vein color Doppler ultrasound. The preoperative and postoperative incidence of DVT was counted and the preoperative to postoperative changes of the DVT was detected. The risk factors related to the preoperative and postoperative DVT were evaluated, including general patient information, operation time, intraoperative blood loss, intraoperative infusion volume, Surgical methods, intraoperative blood transfusion, serological indicators, etc. Single factor analysis was used to screen for risk factors, and then multivariate logistic regression analysis was used to determine the main independent risk factors.Results:The incidence of DVT before and after operation was 36.5%, 57.1%, respectively ( P<0.05). The incidence of unilateral DVT before and after operation was 30.5%, 37.1%, respectively ( P<0.05). The incidence of bilateral DVT before and after operation was 6.0%, 20.0% ( P<0.05). The preoperative rates of proximal, mixed, and distal DVT were 2.8%, 6.8% and 90.4%, respectively. The postoperative rates of proximal, mixed, and distal DVT were 2.2%, 4.7% and 93.1%, respectively. There were 293 patients (60.4%) showing no change in thrombosis, 164 patients (33.8%) aggravation, and 28 patients (5.8%) reduction. Univariate analysis showed the preoperative DVT group and non-DVT group had significant differences in gender, age, time from injury to operation, hemoglobin, hematocrit, D-dimer, previous history of venous thromboembolism (VTE), combination with injuries in other sites ( P<0.05), while not in fracture side, American Society of Anesthesiologists (ASA) score, time from injury to admission, fibrinogen, fracture AO classification, combination with medical disease, and combination with multiple medical diseases ( P>0.05). Postoperative DVT group and non-DVT group were significantly different in gender, operation method, time from injury to operation, and previous history of VTE ( P<0.05), while not in age, fracture side, operation time, intraoperative blood loss, intraoprative blood transfusion, ASA score, time from injury to admission, time from injury to operation, fracture AO classification, combination with medical disease, combination with multiple medical diseases, and combination with injuries in other sites ( P>0.05). Preoperative multivariate logistic regression analysis showed that gender, hemoglobin, hematocrit, previous history of VTE, and combination with injuries in other sites were significantly associated with preoperative DVT ( P<0.05). Postoperative multivariate logistic regression analysis showed gender, time from injury to operation and previous history of VTE were significantly associated with postoperative DVT ( P<0.05). Conclusion:The incidence of DVT is high in patients with intertrochanteric femur fractures, with a significant increase after operation. The type of thrombosis is mainly distal thrombosis and the effect of preventive anticoagulation is not satisfactory. Female, hemoglobin, hematocrit, previous history of VTE, and combination with injuries in other sites are independent risk factors for DVT before operation. Female, time from injury to operation, and previous history of VTE are independent risk factors for DVT after operation.

OBJECTIVETo observe the influence of adding anti-human thymocyte globulin (ATG) into conditioning regimen on graft-versus-host disease (GVHD) and life quality of the patients of allo-peripheral blood stem cell transplantation (PBSCT).

METHODSPatients were distributed into study (19 cases) and control (24 cases) groups at random. Median dose of rabbit ATG was added to the conditioning regimen based on the fludara, busufan and cyclophosphamide (FBC) in study group, and no ATG in the control group. Acute and chronic GVHD disease and Karnofsky scores were compared between two groups after allo-PBSCT.

RESULTSThe patients in the study group received a mean of 6.0 (3 - 9) x 10(8)/kg mononucleated cells and 5.5 (4.5 - 7.5) x 10(6)/kg in the control group. The mean CD(34)(+) cells number was 5.5 (3.0 - 6.5) x 10(5)/kg in the study and 5.0 (3.0 - 7.0) x 10(6)/kg in the control group respectively. Eighteen patients in the study group and in the control group were successfully engrafted. The mean time of absolute neutrophil count recovered more than 500/ micro l was 13 days and 12 days respectively. Acute GVHD occurred in 6 patients of the study group, and 15 of the control group. Seven patients suffered from chronic GVHD and 14 got 90 Kanrofsky scores in a mean of 250 days follow-up in the study group, and 19 patients GVHD and 4 patients respectively in a mean of 440 days follow-up in the control group. There was a significant difference for acute and chronic GVHD and life quality between the two groups.

CONCLUSIONSAddition of anti-thymocyte globulin to the FBC conditioning regimen had no effect on stem cells engraftment but could decrease acute and chronic GVHD and improve patients life quality.

Adolescent ; Adult ; Antilymphocyte Serum ; therapeutic use ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; etiology ; prevention & control ; Humans ; Leukemia ; psychology ; therapy ; Male ; Peripheral Blood Stem Cell Transplantation ; Quality of Life ; T-Lymphocytes ; immunology ; Transplantation Conditioning ; Transplantation, Homologous

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG. The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration-dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2-O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.
Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Ions ; chemistry ; Metals ; chemistry ; Plant Roots ; chemistry ; Polygonaceae ; chemistry ; Stilbenes ; chemistry