In this article we report the results on the synthesis and accumulation of PHAs by Rhodocista pekingensis (strain 3-p), a phototroph that was isolated from wastewater treatment plant. Our results showed that the optimal conditions for PHAs accumulation with strain 3-p were as following: 0.01% Yeast Extract, 0.01% NH 4Cl, Acetate 5 g/L, and medium pH of 7.0～7.2.Under optimized conditions, strain 3-p accumulated PHAs up to 60% of its cellular dry weight. Enzymatic activities of ?-ketothiolase, acetoacetyl-CoA reductase, and PHA polymerase were detected and their activities increased as PHA synthesis initialized. Based on these study, we proposed the metabolic pathway of this strain should be:Acetate (or other fatty acids) - Acetyl-CoA --- thiolase Acetoacetyl-CoA --- reductase D (-)-?-Hydroxybutyryl-CoA ----- PHA polymerase PHAs.

12 isolates of thermoacidophilies were obtained from samples of hot springs of Southern China's Guangdong and Yunna provinces. All isolates are heterotrophic. Cells are rods, Gram positive or variable. The optimal pH and temperature for growth are 3.5-5.5 and 43℃-52℃, respectively. Based on their morphological physiological properties and their 16S rDNA sequences, they were identified as members of Alicyclobacillus.

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.
Animals ; Chemistry, Pharmaceutical ; Diffusion ; Ethanol ; Fatty Alcohols ; Gels ; Injections, Intra-Articular ; Liquid Crystals ; Morphinans ; administration & dosage ; chemistry ; Rats ; Rheology ; Water ; alpha-Tocopherol

In our study, 198 types of pesticides in 120 types 333 lots of traditional Chinese medicine (TCM), which were reasonably classified according to its matrix property, were determined by using the pretreatment platform and gas chromatography-mass spectrometry method. As a result, 158 were contaminated with pesticides. However, the content of pesticides in most TCM was very low. In addition, types of pesticides were different in different part of materia medica. In conclusion, the current status of pesticide residues pollutants in TCM was summarized, and the result can provide proof for the formulation of maximum residue limit. The new species of herbs and the new detecting index should be electively monitored in Chinese Pharmacopeia.
China ; Drug Contamination ; prevention & control ; Pesticide Residues ; analysis ; Plants, Medicinal ; chemistry ; Quality Control

Objective To explore the causes of death and life expectancy after elimination of main causes of disease in residents of Tianjin. Methods The death registry data of Tianjin residents in 2014 were collected and coded in“international classification of disease, 10th edition”. The crude death rate and life expectancy after elimination of main causes of disease were calculated, respectively. Results In 2014, the crude death rate in Tianjin residents was 70.708 per million, while in male and female were 78.728 and 62.637 per million respectively. The main cause of death in Tianjin residents was non-communicable disease. The top four death causes were heart disease, cancer, cerebrovascular disease and respiratory disease, accounting for 31.5%, 23.6%, 22.2% and 8.3% of the total death. The top four life expectancy lost diseases were heart disease, cerebrovascular disease, cancer and respiratory disease, with a 6.46 year, 3.28 year, 3.11 year and 1.25 year life increase respectively. Conclusion Non-communicable diseases are the major reason of death and life expectancy lost disease in Tianjin residents, which needs urgent effective intervention to control.

Objective:To analyze the long-term efficacy and safety of re-irradiation for recurrent glioma.Methods:The data of 52 patients with recurrent gliomas were collected from 2009 to 2019. The median planned targetvolume (PTV) was 73.5 cm 3(49.9-102.7 cm 3) and the median dose was 45.0 Gy (43.0-48.8 Gy). Kaplan-Meier method was used for survival assessment, log-rank test for difference assessment, and Cox’s regression model for multivariate prognostic analysis. Results:The median follow-up time was 32.6 months. The median overall survival (OS) and progression-free survival (PFS) time were 16.1 months (95% CI, 4.1-28.1) and 8.0 months (95% CI, 4.0-12.0). The 1-, 2-and 3-year survival rates were 67%, 43% and 29%, respectively. The 6-month, 1-year and 2-year PFS rates were 67%, 40%, 26%, respectively. Multivariate analysis showed that KPS score and recurrence time significantly affected the OS ( P=0.012, P=0.001). KPS score and time interval between two radiotherapies significantly impacted the PFS ( P=0.003, P=0.018). Stratified analysis showed that KPS score was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅱ initial pathology and reoperation after recurrence ( P<0.001, P=0.012); clinical manifestation was the independent prognostic factor of OS and PFS in patients with WHO grade Ⅲ and Ⅳ initial pathology ( P=0.006, P=0.044). The overall incidence of adverse reactions was 30.8%. Grade 1 adverse reactions accounted for 25.0%, and 5.8% for grade 2. Conclusions:Re-irradiation for recurrent glioma yields good long-term clinical efficacy and tolerable adverse reactions.

Objective To report the phototoxicity effects of a novel photosensitizer ZnPcS4-BSA on photodynamic therapy (PDT) towards human U251 glioma cells in vitro. Methods The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra to determine the optimal incubation time. Human U251 glioma cells were incubated with ZnPcS4-BSA of various concentrations and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay.Flow cytometer was used to detect apoptosis.Gene expressions of vascular endothelial growth factor (VEGF) were detected by Real-Time PCR in the U251 cells after PDT and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results The uptake of ZnPcS4-BSA by U251 glioma cells reached the maximum after incubation for 4 hours.ZnPcS4-BSA of different concentrations without laser irradiation had no significant effects on cell survival rates (P＞0.05).Without ZnPcS4-BSA incubation,compared with 0,25,50,100,200 J/cm2 groups, the cell survival rate of the 400 J/cm2 group was significantly lower (P＜0.05), whereas no significant difference was found between any other two groups. When the U251 glioma cells incubated with 30 μ mol/L ZnPcS4-BSA for 4 hours underwent laser irradiations of 25,50,100,200 J/cm2,the cellular survival rates significantly decreased with the increased energy densities (P＜0.05). When the U251 glioma cells incubated with ZnPcS4-BSA of 20,40,60,80,100 μ mol/L for 4 hours underwent laser irradiation of 200 J/cm2, the cellular inhibition rates significantly increased with the increased concentrations (P ＜0.05). Compared with controls, the cellular apoptosis and VEGF expression significantly increased in the U251 glioma cells incubated with ZnPcS4-BSA of 20 μmol/L after laser irradiation of 100 J/cm2 (P＜0.05). Conclusion The novel ZnPcS4-BSA is a good photosensitizer for PDT towards U251 glioma cells,because the ZnPcS4-BSA-mediated PDT can induce effective apoptosis of the targeted cells.

OBJECTIVETo investigate the effect of lithium chloride (LiCl) on cell cycle of HK-2 cells and explore the possible pathways involved.

METHODSHK-2 cells were treated with LiCl at different concentrations (5, 12.5, 20, and 25 mmol/L) for 12, 24, 48, or 72 h, and the changes in cell cycle and viability were detected using flow cytometry and CCK-8 assay, respectively. Western blotting was used to analyze the changes in the expressions of cyclin B1 and CDK1 (the two G2 phase-related proteins) and those of AKT/GSK-3β signaling pathway-related proteins in the treated cells.

RESULTSLiCl treatment time- and concentration-dependently increased HK-2 cell percentage in G2 phase and decreased the cell vitality. The expressions of cyclin B1, CDK1, p-GSK-3β, and β-catenin increased and the expression of p-AKT decreased significantly in the cells as LiCl treatment time and concentration increased.

CONCLUSIONLiCl may cause HK-2 cell cycle arrest in G2 phase through activation of the AKT/GSK-3β signaling pathway.

To investigate the effects of human anti-BAFF scFv-Fc against the hsBAFF, ICR mice were randomly divided into six groups: control, hsBAFF (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (2 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + human IgG (1 mg x kg(-1)) and hsBAFF (1 mg x kg(-1)) + human IgG (2 mg x kg(-1)) groups. The effects of scFv-Fc administration on the proliferation of B lymphocytes were evaluated using an MTT assay. The titres of antibody in the serum and B lymphocytes differentiation were assessed by ELISA and flow cytometry, respectively. The results showed that administration of scFv-Fc to mice injected with hsBAFF significantly prevented human BAFF-induced increases in splenic B cell numbers and serum immunoglobulin levels. Furthermore, this fully human antibody would avoid inducing the human anti-mouse antibody (HAMA) response when used in humans. These findings suggest that the compact antibody may be useful in therapeutic or diagnostic application of the BAFF-associated autoimmune diseases in human.
Animals ; B-Cell Activating Factor ; immunology ; metabolism ; B-Lymphocytes ; cytology ; Body Weight ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Female ; Humans ; Immunoglobulin Fc Fragments ; immunology ; metabolism ; Immunoglobulin G ; blood ; immunology ; Immunoglobulin M ; blood ; Mice ; Mice, Inbred ICR ; Random Allocation ; Recombinant Fusion Proteins ; immunology ; metabolism ; Single-Chain Antibodies ; immunology ; metabolism ; Spleen ; cytology

Aim To observe the effects of realgar nano-particles on B cell non-Hodgkin's lymphoma Raji cells in vitro. Methods Realgar nanoparticles and crude realgar particles were characterized with a laser particle size analyzer, a transmission electron microscopy (TEM)and an atomic force microscopy(AFM). The morphological changes of proliferation of Raji cells brought about by the use of realgar naoparticles and crude realgar particles were observed with a light mi-croscope. The membrane changes of Raji cells treated with realgar naoparticles and crude realgar particles were observed with AFM. The ultrastructures of Raji cells were observed with TEM. The inhibitory effects of Raji cells treated with realgar naoparticles and crude realgar particles were measured with MTT. The nuclear apoptosis morphologies of Raji cells were observed with fluorescence microscopy. The apoptosis rates and the cell cycle distributions of Raji cells treated with real-gars were measured with flow cytometry. Results The size of realgar nanoparticles and crude realgar particles was (79 ± 8)nm and (1. 89 ± 0. 2)μm,respectively. Light microscopy showed that realgar nanoparticles could inhibit the aggregation growth of Raji cells. AFM showed that Raji cells treated with realgar nanoparticle became shrank, had smaller volume and lost the growth state of stretching out. Raji cells treated with crude realgars did not change significantly. TEM showed Raji cells treated with realgar nanoparticle had damaged subcellular organelles and mitochondria with increased vacuoles. The Raji cells treated with crude realgar did not change significantly. MTT assay showed that when treated with the final concentration of 50 mg ·L - 1 of realgar nanoparticle for 24 h,the cell survival rate of Raji cells was (40 ± 2)% . When treated with the same concentration of crude realgar,its survival rate was (65 ± 3)% . When treated with 50 mg·L - 1 of realgar nanoparticle for 48 h,its survival rate was only 10 % ,and when treated with crude realgar ,its survival rate was (42 ± 2 )% . Fluorescence micro-scope indicated that the Raji cells treated with realgar nanoparticle had obvious nuclear apoptosis,which was not obvious in crude realgar group. Flow cytometry showed that the total apoptosis rate of Raji cells in-duced by realgar nanoparticles and by crude realgar was 11. 14%,15. 9%,respectively. Compared with those treated with crude realgar,the Raji cells treated with realgar nanoparticles presented a significantly higher ratio cell distribution in G1 phase and an obvious decreased ratio in S phase. Conclusion Compared with crude realgar particles,the same dose of realgar nanoparticles can significantly inhibit the proliferation of Raji cells,destroy their sub-cellular structure,and induce the cell apoptosis of Raji cells.