Activated mTOR signaling pathway in myofibers with inherited metabolic defect might be an evidence for mTOR inhibition therapies / 中华医学杂志(英文版)

BACKGROUND@#Abnormally activated mechanistic target of rapamycin (mTOR) pathway has been reported in several model animals with inherited metabolic myopathies (IMMs). However, the profiles of mTOR pathway in skeletal muscles from patients are still unknown. This study aimed to analyze the activity of mTOR pathway in IMMs muscles.@*METHODS@#We collected muscle samples from 25 patients with mitochondrial myopathy (MM), lipid storage disease (LSD) or Pompe disease (PD). To evaluate the activity of mTOR pathway in muscle specimens, phosphorylation of S6 ribosomal protein (p-S6) and p70S6 kinase (p-p70S6K) were analyzed by Western blotting and immunohistochemistry.@*RESULTS@#Western blotting results showed that p-p70S6K/p70S6K in muscles from LSD and MM was up-regulated when compared with normal controls (NC) (NC vs. LSD, U = 2.000, P = 0.024; NC vs. MM: U = 6.000, P = 0.043). Likewise, p-S6/S6 was also up-regulated in muscles from all three subgroups of IMMs (NC vs. LSD, U = 0.000, P = 0.006; NC vs. PD, U = 0.000, P = 0.006; NC vs. MM, U = 1.000, P = 0.007). Immunohistochemical study revealed that p-S6 was mainly expressed in fibers with metabolic defect. In MM muscles, most p-S6 positive fibers showed cytochrome C oxidase (COX) deficiency (U = 5.000, P = 0.001). In LSD and PD muscles, p-S6 was mainly overexpressed in fibers with intramuscular vacuoles containing lipid droplets (U = 0.000, P = 0.002) or basophilic materials (U = 0.000, P = 0.002).@*CONCLUSION@#The mTOR pathway might be activated in myofibers with various metabolic defects, which might provide evidence for mTOR inhibition therapy in human IMMs.

Clinical, Neuroimaging, and Pathological Analyses of 13 Chinese Leigh Syndrome Patients with Mitochondrial DNA Mutations / 中华医学杂志(英文版)

Background@#Leigh syndrome (LS) is a rare disease caused by mitochondrial defects and has high phenotypic and genotypic heterogeneity. We analyzed the clinical symptoms, neuroimaging, muscular histopathology, and genotypes of 13 Chinese LS patients with mitochondrial DNA (mtDNA) mutations.@*Methods@#Mutations in mtDNA were identified by targeted sequencing. The brain imaging features on magnetic resonance imaging (MRI) were analyzed. The levels of lactate in fasting blood and cerebrospinal fluid (CSF) were routinely tested. The levels of urinary organic acids, plasma amino acids, and acylcarnitines were examined with gas chromatography-mass spectrometry and tandem mass spectrometry. The histopathological traits of skeletal muscles were analyzed under microscope.@*Results@#Among 13 patients, mutations of MT-NDs (n = 8) and MT-ATP6 (n = 4) genes were most common. Strabismus (8/13), muscle weakness (8/13), and ataxia (5/13) were also common, especially for the patients with late-onset age after 2 years old. However, respiratory distress was common in patients with early-onset age before 2 years old. The most frequently affected brain area in these patients was the brain stem (12/13), particularly the dorsal part of midbrain, followed by basal ganglia (6/13), thalamus (6/13), cerebellum (5/13), and supratentorial white matter (2/13). Besides, the elevated lactate levels in CSF (6/6) were more common than those in serum (7/13). However, the analysis of abnormal plasma amino acid and urinary organic acid showed limited results (0/3 and 1/4, respectively). Muscular histopathology showed mitochondrial myopathy in the three late-onset patients but not in the early-onset ones.@*Conclusions@#Noninvasive genetic screening is recommended for mtDNA mutations in MT-NDs and MT-ATP6 genes in patients with ophthalmoplegia, muscle weakness, ataxia, and respiratory disorder. Furthermore, the lactate detection in CSF and the brain MRI scanning are suggested as the diagnosis methods for LS patients with mtDNA mutations.

Preliminary study on biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the biological characteristics of CD(44)(+) stem cells in human laryngeal carcinoma.</p><p><b>METHODS</b>Tumor samples were obtained from 5 patients, and then the human laryngeal carcinoma cells were cultured in vitro by primary tissue culture technique. Taking CD44 molecule as a marker to isolate CD(44)(+) subpopulation cells from laryngeal carcinoma cells for further study. CD(44)(+) and CD(44)(-) cells were cultured and observed fex their development. CD(44)(+) and CD(44)(-) cells were compared in their functional status (mRNA), cell cycles (G0/G1), their degree of differentiation (CK14 and Involucrin expression) and their morphologic character of the clone.</p><p><b>RESULTS</b>The percentages of CD(44)(+) cells were about 49.8% approximately 53.5% and the median was 51.3%. After culturing CD(44)(+) cells isolated from laryngeal carcinoma could proliferate and the percentage of CD(44)(+) remained the same. CD(44)(+) tumor cells contained much less RNA, more G0/G1 cells, expressed more CK14 protein and less Involucrin protein (less differentiated state). CD(44)(+) cells were multangular in shape with protuberances; CD(44)(-) cells showed a sharp and spindle feature. In comparison with CD(44)(-) cells, CD(44)(+) cells could create heterogeneous offspring by single cell culture of limiting dilution. By observing clone forming rate after single cell planting, it was found that the CD(44)(+) cells had stronger proliferation ability.</p><p><b>CONCLUSIONS</b>CD(44)(+) cells possess some characteristics of stem cells, laryngeal carcinoma stem cells maybe exist in CD(44)(+) cells.</p>

Microstructure of novel solid lipid nanoparticle loaded triptolide / 药学学报

Novel solid lipid nanoparticle (SLN) system is prepared with Compritol ATO 888 and tricaprylic glyceride. DSC, XRD, SAXS and NMR are employed to study the novel carrier property and microstructure. When the peak melting point decreased from 70.8 degrees C to 61.4 degrees C, the enthalpy sharply decreased. It could be concluded that the regular crystal lattices in the novel carriers are broken out for the oil joined in them. Melting behavior is occurred at -17.7 degrees C while novel SLN is composed of oil and solid lipid mixture from the DSC measurement. Most alpha phase and least beta' phase are in the nano carrier system whether drug loading or not from the XRD investigation. There is only 0.1 nm change of long space among the novel SLN made of mixture and the lipid matrix and traditional SLN; therefore, it is impossible of the oil molecular insert into the solid glyceride structure. Since the different melting behavior (DSC measurements) and molecular move state (NMR investigations), two lipid matrix are still in two state of liquid and solid lipid in the novel SLN carrier. Presume the microstructure of the novel SLN prepared by our experiment would be that liquid oil has formed superfine nano accommodation encapsulated with solid lipid, but the whole particle is still in nano size range.

Isopropyl myristate molecular gels and drug-loaded transdermal capability / 药学学报

<p><b>AIM</b>To prepare of isopropyl myristate (IPM) molecular gels and investigate of its transdermal capability.</p><p><b>METHODS</b>Microstructure of IPM gels was studied by scanning electron microscope (SEM) and optical microscope (OM). The rheology and thixotropy of IPM gels were investigated by viscosity. Triptolide was used as model drug to investigate its transdermal capability.</p><p><b>RESULTS</b>The microstructure of IPM gels was a three-dimension network formed by the aggregation of Span 60 in IPM, which was rod-like tubular aggregate. It has good rheology and thixotropy. There was a good linear correlation between the accumulative permeated amount per unit area and the time for triptolide-loaded IPM gels. The permeation process agreed with zero order pharmacokinetics. The average permeability through rat skin for triptolide was 19.26 ng x cm(-2) x h(-1), which was 2.92 times of triptolide unguents obtained commercially available.</p><p><b>CONCLUSION</b>Isopropyl myristate molercular gel can be formed by span 60 assemblies. Transdermal capability drug-loaded IPM gels was better than that of triptolide unguents.</p>

Progress in research on triptolide / 中国中药杂志

To further understand triptolide, this paper has introduced the pharmacology, pharmacokinetics, toxicity, the clinic application and semi-synthesis of triptolide on basis of importance and significant contents of reference which have been consulted in the past twenty years. Presently triptolide and Tripterygium wilfordii have been a hot spot of modernization of Chinese traditional medicine. It is very important to develop a new dosage form of high effect and low toxicity by making use of advanced technology according to its characteristics.

Preparation and toxicity of triptolide-loaded poly (D,L-lactic acid) nanoparticles / 药学学报

<p><b>AIM</b>Investigations on reducing the toxicity of triptolide through poly(D, L-lactic acid) nanoparticles as a drug carrier by oral administration to Wistar rats.</p><p><b>METHODS</b>Triptolide-loaded poly (D, L-lactic acid) nanoparticles (TP-PLA-NPs) were prepared by modified spontaneous emulsification solvent diffusion (modified-SESD). The shape of nanoparticles was observed by transmission electron microscope (TEM). The size distribution and mean diameter were measured by laser light scattering technique. The entrapment efficiency and contents of drug loading were determined by RP-HPLC. The physical state of drug loaded in nanopartiles were primarily investigated by X-ray powder diffractometry. TP-PLA-NPs release behavior in vitro was carried out. After oral administration of the nanoparticles to Wistar rats in 15d, the toxicity for liver and kidney were studied by determining aspartate transaminase (AST), alanine transaminase (ALT) and blood urea nitrogen in serum and concentration of protein in urine.</p><p><b>RESULTS</b>The preparation process adapted to the formulation was as follows: the volume ratio of the aqueous and organic phases was 40/15; the surfactant concentration was 1%; the drug concentration was 0.3%; triptolide-PLA was 1:15 (w/w). The mean diameter was 149.7 nm and the polydispersity index was 0. 088 for the nanoparticles prepared by above conditions. The entrapment efficiency and content of drug loading were 74.27% and 1.36%, respectively. The release behavior of drug in vitro showed an initial burst effect, subsequently a slower rate stage. The results indicated that the liver toxicity (P < 0.01) and kidney toxicity (P < 0.05) caused by triptolide could be decreased significantly by nanoparticles carrier.</p><p><b>CONCLUSION</b>PLA-NPs might be used as a new oral carrier for triptolide.</p>

Characterization of Me. PEG-PLA copolymer nanoparticles prepared by modified spontaneous emulsion-solvent evaporation / 药学学报

<p><b>AIM</b>Characterization of poly (D, L-lactic acid)/monomethylether terminated/polyethylene glycol (Me. PEG-PLA) block copolymers nanoparticles.</p><p><b>METHODS</b>Me. PEG-PLA block copolymers were prepared by bulk polymerization. A series of nanoparticles were made from Me. PEG-PLA block copolymer by modified spontaneous emulsion-solvent evaporation technique.</p><p><b>RESULTS AND CONCLUSION</b>The structure of copolymer was performed by means of 1HNMR and FT-IR. The morphological examination of nanoparticles was performed by means of atomic force microscope (AFM). Results indicated that nanoparticles exhibited a smooth spherulite and core-shell structure. The hydrophilic shell is consisted of PEG segments and hydrophobic core is consisted of PLA segments. Zeta potential of nanoparticles was zero and further indicated core-shell structure. The particle size and size distribution of nanoparticles were measured by laser light scattering technique. The effective particle size range was from 70 to 160 nm and showed a normal distribution.</p>

Influence of pulmonary metastasis of three bisdioxopiperazines on Lewis lung carcinomas in vivo / 中国药理学通报

AIM To compare the effects of probimane( Pro), bimolane ( Bim) and razoxane( Raz) on animal tumor metastasis in vivo . METHODS A biological inoculation method for assessment of pulmonary metastasis of Lewis lung carcinoma(3LL) was employed. RESULTS Pro and Bim inhibited the pulmonary metastasis of 3LL both from d 2 and from d 8 injections, but Raz only inhibited the pulmonary metastasis of 3LL from d 2 injections. Pro inhibited the pulmonary metastasis of 3LL more potently than Bim did at equitoxic dosage. CONCLUSION Pro is better in the treatment of pulmonary metastasis of 3LL than Raz for its possible novel molecular mechanisms.