The colorimetric method has been well developed to detect the sensitivity of most antiproliferative drugs with the advantages of objectivity and accuracy. Many parameters including the dosage and the absorbance of crystalline violet, the extraction time of the solvent, plate effect and marginal effect were investigated, then the optimized method was applied further to measure the biological activity during the refolding of recombinant human IFN-? inclusion bodies.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
Anti-Inflammatory Agents ; pharmacology ; Carthamus tinctorius ; Computational Biology ; Drugs, Chinese Herbal ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Humans ; Inflammation ; drug therapy ; Medicine, Chinese Traditional

A novel multi-components microcapsule--SA/CS-CaCl2/PMCG system was introduced. The effects of PMCG and SA/CS-CaCl2/PMCG microcapsules on the growth of free E. coli and Saccharomyces cerevisiae were studied respectively. In addition, the growth of immobilized E. coli and Saccharomyces cerevisiae were also investigated. The results showed that: Just like other synthetic polycations, PMCG above certain concentration (0.5%) strongly inhibited the growth of free E. coli and Saccharomyces cerevisiae, but SA/CS-CaCl2/PMCG microcapsules almost had no effects on their growth and on the consumption of glucose concentration by Saccharomyces cerevisiae. What's more, immobilized E. coli and Saccharomyces cerevisiae grew almost as normally as free cultivation. As a whole, SA/CS-CaCl2/PMCG microcapsules had good biocompatiability and can be used as a new immobilization system.
Alginates ; toxicity ; Biocompatible Materials ; toxicity ; Calcium Chloride ; toxicity ; Cellulose ; analogs & derivatives ; toxicity ; Escherichia coli ; drug effects ; growth & development ; Glucuronic Acid ; Hexuronic Acids ; Polymers ; toxicity ; Saccharomyces cerevisiae ; drug effects ; growth & development

The 11-hydroxylation of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione as a useful intermediate for the preparation of hormones can be achieved by the mycelium of Absidia coerulea at higher conversion rate than using other strains. In this paper 16alpha,17alpha-epoxy-4-pregenene-3,20-dione mixed with a little water, beta-cyclodextrin, Tween-80 was introduced into the fermentation broth after ultrasonication to increase pseudo-water-solubility of the hydrophobic substrate. This pseudo-crystallo feed could avoid the toxicity of organic solvents and was more available for the microbial transformation. The multi layer feed-forward neural network was used to setup a model which indicated the relationship between medium and feed components and the conversion rate. Particle swarm optimization (PSO), which was a stochastic global optimization algorithm and of which the convergence speed was high, was applied to obtain the optimal concentration of the medium and feed components. At optimum conditions with the pseudo-crystallo feed, the conversion rate of 16alpha,17alpha-epoxy-4-pregenene-3,20-dione at an initial concentration of 10 g/L was 87.5% in shaking flasks. The conversion rate of the substrate was up to 86.6% at higher concentration of 20 g/L feed in a 3.7 L fermentor.
Absidia ; metabolism ; Fermentation ; Hydroxylation ; Pregnenediones ; metabolism

Disulfide bond formation protein A, DsbA, is one of the important proteins located in E. coli periplasm, which is a foldase facilitating the folding of nascent secreted proteins, especially for those with many pairs of disulfide bonds. The crystal structure and phylogenetic analysis of DsbA and DsbA-mediated protein folding, alternatively in vivo and in vitro, are summarized. Both the extremely low pKa of Cys30 , about 3.5, and the destabilizing effect of the active site disulfide contribute to its strong oxidizing power. The Cys30 is also considered as the most important residue closely related to its activity using site-directed mutagenesis methodology. DsbA could effectively assist proteins folding, both in vivo coexpressed with the target protein, and in vitro replenished as foldases. Moreover, DsbA also has the chaperone-like activity in the assistant refolding of genetically engineered inclusion bodies.
Amino Acid Sequence ; Disulfides ; chemistry ; metabolism ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; chemistry ; classification ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Disulfide-Isomerases ; chemistry ; classification ; metabolism ; Protein Folding ; Protein Structure, Tertiary ; Sequence Homology, Amino Acid

Tumor brings great threat to human public health. In recent years, incidence rate and mortality of tumor were rapidly increased in the world. Anti-tumor therapies have undergone the development of cytotoxic therapy, targeted therapy, and immunotherapy. Among them, tumor immunotherapy is rapidly developed and becomes an important anti-tumor therapy in recent years, although it also brings some related side effects. Tumor microenvironment (TME) is composed of immune cells, vascular vessels, fibroblasts, the extracellular matrix, etc. TME significantly affects the efficacy of immunotherapy. Macrophages in the TME are named as tumor associated macrophages (TAMs). Recently, increasing studies have shown that TAMs play an important role in the regulation of tumor immunity, especially in tumor immune surveillance and immune escape. Currently, more and more anti-tumor immunotherapy strategies targeting TAMs are at the development stage. Based on the important role of TAMs in the TME and their potential as therapeutic targets in tumor immunotherapy, we first reviewed the subtypes and functions of TAMs, as well as the roles of TAMs in tumors. Furthermore, we summarized the research progress on anti-tumor strategies targeting TAMs and the current status of drug targeting TAMs. The current review will provide new ideas and novel insights for tumor immunotherapy.

OBJECTIVETo report the clinical, genetic, and molecular characterization of two Chinese families with Leber's hereditary optic neuropathy (LHON).

METHODSOphthalmological examinations showed that only probands in two families exhibited visual loss at the age of 10 and 17 years respectively. The entire mitochondrial genome of two probands was PCR amplified in 24 overlapping fragments using sets of oligonucleotide primers.

RESULTSMutational analysis of mitochondrial DNA (mtDNA) in these pedigrees revealed the absence of three common LHON associated G11778A, G3460A and T144484 mutations but the presence of homoplastic LHON associated ND4 G11696A mutation, which was present in one out of 167 Chinese healthy controls.

CONCLUSIONSequence analysis of the complete mitochondrial genomes in two pedigrees showed the distinct sets of mtDNA polymorphisms, belonging to Eastern Asian haplogroup D4. The incomplete penetrance of visual loss and the presence of one in 167 controls suggested that this mutation itself is insufficient to produce a clinical phenotype and other modifier factors play a role in the phenotypic manifestation. The lack of functional mtDNA variants in these pedigrees ruled out the role of mitochondrial background in the phenotypic expression of visual loss. Therefore, nuclear modifier gene(s) or environmental factor(s) may play a role in the phenotypic expression of the LHON-associated G11696A mutation in two Chinese pedigrees.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; DNA, Mitochondrial ; genetics ; Family ; Female ; Humans ; Male ; Mutation ; Optic Atrophy, Hereditary, Leber ; genetics ; Pedigree ; Phenotype

Objective: To analyze associations of MRI-lesions and clinical features with disability in patients with multiple sclerosis (MS) in Shanghai, China. Methods: We studied patients with MS, identified from a survey in Shanghai, whose sites of lesions in the CNS was based on the MRI examinations. Associations between MRI-lesions, various clinical variables and the severity of disability were analyzed with univariate and multivariate logistic regression analysis. Results: There were 210 patients in this study. The disability of the patients with lesions confined to the spinal cord was significantly more severe than those with lesions in the brain (p < 0.008). Current age (OR: 1.041, 95% CI: 1.007~1.077), MS duration (OR: 1.082, 95% CI: 1.011~1.159) and MRI-lesions in the spinal cord (OR: 2.441, 95% CI: 1.039~5.737) were significantly associated with severity of disability on multivariate logistic regression analysis. Conclusion: MRI-lesions in the spinal cord, older age, a longer MS duration were significantly associated with a more severe disability in this MS study in Shanghai China.

Leber's hereditary optic neuropathy (LHON) is a maternally inherited disorder leading to rapid, painless, bilateral and usually permanent central vision loss in young adults, males are preferentially affected. The maternal transmission of this visual dysfunction in LHON families suggested that mutations in the mitochondrial DNA (mtDNA) are the molecular bases of the disorder. The ND1 G3460A, ND4 G11778A and ND6 T14484C mutations in the genes encoding the subunits of respiratory chain complex I, account for more than 50% of LHON families worldwide. These three mutations are designated to be primary mutations because they impart a high risk for LHON expression. However, matrilineal relatives within and among families, despite carrying the same LHON-associated mtDNA mutation(s), exhibit a wide range of onset, severity, and the progression of visual impairment. These findings strongly indicated that the LHON-associated primary mutation(s) are the primary factors underlying the development of vision loss, but they themselves are insufficient to produce a clinic phenotype. The prone to male, incomplete penetrance, and phenotypic variability of vision loss suggest that other modifier factors including personal factors, environmental factors, nuclear modifier genes and mitochondrial haplotypes contribute to the phenotypic expression of these mtDNA mutations. In particular, the mitochondrial haplotypes may play a synergic role in the development of vision loss in the families carrying the LHON-associated primary mtDNA mutation(s).
DNA, Mitochondrial ; genetics ; Genome, Human ; genetics ; Genomics ; Haplotypes ; Humans ; Mitochondria ; genetics ; Optic Atrophy, Hereditary, Leber ; genetics ; pathology

OBJECTIVETo study the paracrine effect of the neuroendocrine differentiation cells and its influence on androgen receptor expression of prostate carcinoma cells.

METHODSEstablished an in vitro induced model of neuroendocrine differentiation of prostate carcinoma (LNCaP(NE), PC-3M(NE)), and investigate the proliferation effect of neuroendocrine phenotype cells (LNCaP(NE), PC-3M(NE)) on other non-neuroendocrine phenotype cells (LNCaP, PC-3M) by feeding non-neuroendocrine phenotype cells with the medium pre-incubated with induced neuroendocrine phenotype cells. It was also tested the regulation of androgen receptor mRNA and androgen receptor protein expression of LNCaP(NE) cells on LNCaP cells by reverse transcriptase polymerase chain reaction and Western Blot in the presence or absence of androgens.

RESULTSThe medium pre-incubated with PC-3M(NE) cells could promote the proliferation of PC-3M cells. The medium pre-incubated with LNCaP(NE) cells could promote the proliferation of LNCaP cells, and reduce the expression of androgen receptor in the latter in the absence of androgens, but the negative results were observed in the presence of androgens.

CONCLUSIONSThe neuroendocrine phenotype cells of prostate cancer can reduce the expression of androgen receptor in prostate cancer cells and promote them to proliferate by means of paracrine in the blockade of androgens.

Androgens ; pharmacology ; Cell Division ; drug effects ; Cell Line, Tumor ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Receptors, Androgen ; metabolism