The in vivo Therapeutic Effect of Free Wanderer Powder (逍 遙 散 xiāo yáo sǎn, Xiaoyaosan) on Mice with 4T1 Cell Induced Breast Cancer Model

ABSTRACTIn the present study, we investigated the therapeutic effect of a classical TCM formula, Free Wanderer Powder (逍遙散 xiāo yáo sǎn), in a breast cancer mouse model induced with estrogen-insensitive breast cancer 4T1 cells. Ovariectomized Balb/c mice (6-8 weeks) or sham mice were injected into the fourth mammary fat pad with 4T1 cells in which tumors were palpable 7days after injection. On the eighth day, the mice were divided into 4 groups and tubefed daily with vehicle, Free Wanderer Powder (逍遙散 xiāo yáo sǎn) formula or tamoxifen for 28days. Tumor growth inhibition and the decrease of the average tumor mass were most evident in mice treated with Free Wanderer Powder (逍遙散 xiāo yáo sǎn). Free Wanderer Powder (逍遙散 xiāo yáo sǎn) treatment significantly reduced Bcl-2 and elevated Bax and p53 protein expressions in breast cancer tumor. These results were further confirmed by immunohistochemisty. Tamoxifen could decrease spleen mass and Bcl-2 protein expression, increase the Bax protein expression as well as exert uterotrophic effects by increasing uterus index and inducing the gene expressions in the uterus. Taken together, these results show that Free Wanderer Powder (逍遙散 xiāo yáo sǎn) treatment induced apoptosis at protein level and inhibited the tumor growth in 4T1-induced ovariectomized Balb/c female mice, indicating the possibility of its future use for treatment of estrogen-insensitive breast caner

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Biodegradable nanoparticles of amphiphilic triblock copolymers based on poly(3-hydroxybutyrate) and poly(ethylene glycol) as drug carriers

New amorphous amphiphilic triblock copolymers of poly(3-hydroxybutyrate)–poly(ethylene glycol)–poly(3-hydroxybutyrate) (PHB–PEG–PHB) were synthesized using the ring-opening copolymerization of β-butyrolactone monomer. They were characterized by fluorescence, SEM and 1H NMR. These triblock copolymers can form biodegradable nanoparticles with core–shell structure in aqueous solution. Comparing to the poly(ethylene oxide)–PHB—poly(ethylene oxide) (PEO–PHB–PEO) copolymers, these nanoparticles exhibited much smaller critical micelle concentrations and better drug loading properties, which indicated that the nanoparticles were very suitable for delivery carriers of hydrophobic drugs. The drug release profile monitored by fluorescence showed that the release of pyrene from the PHB–PEG–PHB nanoparticles exhibited the second-order exponential decay behavior. The initial biodegradation rate of the PHB–PEG–PHB nanoparticles was related to the enzyme amount, the initial concentrations of nanoparticle dispersions and the PHB block length. The biodegraded products detected by 1H NMR contained 3 HB monomer, dimer and minor trimer, which were safe to the body

Role of Physiotherapy in Preventing Failure of Primary Anterior Cruciate Ligament Reconstruction

Background/Purpose: Anterior cruciate ligament (ACL) reconstruction is routinely performed in sports medicine. We aimed to determine if there is any protective effect of postoperative physiotherapy in preventing graft rupture after primary ACL reconstruction (ACLR). Methods: A retrospective case–control study was carried out, with demographic data, concomitant meniscal injury, and intraoperative fixation methods matched. The number of sessions of physiotherapy attended by the rupture group and nonrupture group were compared using binary logistic regression. Results: No significant relationship between the frequency of postoperative physiotherapy and occurrence of graft rupture after primary ACLR was identified. Conclusion: Further research is needed to verify the effect of physiotherapy in the prevention of graft rupture after primary ACLR

In vitro cytotoxicity, hemolysis assay, and biodegradation behavior of biodegradable poly(3-hydroxybutyrate)–poly(ethylene glycol)–poly(3-hydroxybutyrate) nanoparticles as potential drug carriers

Nanoparticles based on amorphous poly(3-hydroxybutyrate)–poly(ethylene glycol)–poly(3-hydroxybutyrate) (PHB-PEG-PHB) are potential drug delivery vehicles, and so their cytotoxicity and hemolysis assay were investigated in vitro using two kinds of animal cells. The PHB-PEG-PHB nanoparticles showed excellent biocompatibility and had no cytotoxicity on animal cells, even when the concentrations of the PHB-PEG-PHB nanoparticle dispersions were increased to 120 μg/mL. Moreover, no hemolysis was detected with the PHB-PEG-PHB nanoparticles, suggesting that the PHB-PEG-PHB nanoparticles were obviously much hemocompatible for drug delivery applications. In the presence of intracellular enzyme esterase, the biocompatible PHB-PEG-PHB nanoparticles might be hydrolyzed, and their biodegradable behavior was monitored by the fluorescence spectrum and the pH meter. The initial biodegradation rate of the PHB-PEG-PHB nanoparticles was closely related to the enzymatic amount and the PHB block length. Compared with that obtained from the fluorescence determination, the initial biodegradation rate from pH measurement was faster. The biodegraded products mainly consisted of 3HB monomer and dimer, which were the metabolites present in the body