Development and validation of size exclusion column-high performance liquid chromatography for purity determination of bulk material of freeze-dried rabies vaccine for human use / 中国生物制品学杂志

@#Objective To develop and validate size exclusion column-high performance liquid chromatography(SEC-HPLC)for determination of the purity of bulk material of freeze-dried rabies vaccine for human use.Methods Chromatography column TSK-gel G6000PW_(XL)(7.8 mm × 30 cm,13 μm)was used for the determination(column temperature 30 ℃)with mobile phase of 0.1 mol/L PB buffer(pH 7.8)at a flow rate of 0.5 mL/min.The detection wavelength was 280 nm and injection volume was 20 μL.The method was validated for system suitability,specificity,precision and durability and determined for detection limit and quantitation limit,which was applied to analyze bulk material purity of freeze-dried rabies vaccine for human use of 3 batches of Vero cells and 1 batch of human diploid cells.Results The resolution of target protein spectrum peak of bulk material of reference sample and freeze-dried rabies vaccine for human use prepared with two substrates was more than 1.5 with a tailing factor less than 1.5;The blank solvent showed no absorption peak at the position of target protein peak with no interference in the determination;The RSDs of retention time and peak area in precision verification were both less than 2.0%;The quantitative limit was 10 μg/mL,and the detection limit was 4 μg/mL;The reference sample was injected three times continuously at three different detection wavelengths of 278,280 and 282 nm,and the RSDs of retention time and peak area were also less than 2.0%.The purity of 4 batches of freeze-dried rabies vaccine bulk material for human use was all more than 97%.Conclusion The developed SEC-HPLC for determination of the purity of freeze-dried rabies vaccine bulk material for human use showed good specificity,precision and durability,which provided a reliable method for the quality control of human rabies vaccine.

Research process of reactive oxygen species-based tumor immunomodulation / 药学学报

Reactive oxygen species (ROS) is defined as the electron reduction product of oxygen with high reactivity which can maintain normal physiological functions and redox homeostasis. The tumor microenvironment is in a state of oxidative stress. ROS can affect multiple processes of tumor immune response by modulating the phenotype and functions of tumor cells and immune cells. With the rapid development of immunology, ROS-based tumor immunomodulation has been widely concerned and studied. In this review, the mechanism of ROS participating in tumor immune response is elaborated. Meanwhile, the research process and application of ROS in tumor immunomodulation in recent years are reviewed and analyzed.

Effect of different parts of Pinelliae Rhizoma Decoction against airway inflammation and analysis of effective components / 中国中药杂志

This study investigated the material basis and mechanism of Pinelliae Rhizoma Decoction in the treatment of airway inflammation. The cigarette smoke combined with lipopolysaccharide(LPS) was used to induce an airway inflammation model in mice. The expression levels of IL-6 and IL-8 in the bronchoalveolar lavage fluid(BALF) and the phosphorylation levels of p38 and IκB in the lungs of mice were taken as indexes to screen the effective extracts by system solvent extraction from Pinelliae Rhizoma Decoction(dichloromethane extract, ethyl acetate extract, n-butanol extract, etc.). Meanwhile, the human bronchial epithelial(16-HBE) cell model of cigarette smoke extract(CSE)-induced injury was established, and the mRNA expression levels of IL-6 and IL-8 and the phosphorylation levels of p38 and IκB proteins were also taken as indexes to evaluate the anti-inflammatory effect of different extracts of Pinelliae Rhizoma Decoction. The results showed that Pinelliae Rhizoma Decoction significantly antagonized airway inflammation in mice by down-regulating the expression levels of IL-6 and IL-8 in mice with airway inflammation and 16-HBE cells with CSE-induced injury and inhibiting the phosphorylation levels of p38 and IκB. The dichloromethane and ethyl acetate extracts of Pinelliae Rhizoma Decoction showed significant anti-inflammatory effects, while such effects of other extracts were not prominent. Furthermore, the database of Pinelliae Rhizoma composition was constructed, and the components in effective extracts were analyzed by HPLC-TOF-MS and Nano-LC-MS/MS. As revealed by the results, the compositions of the two effective extracts were similar with 36 common components. They were combined and then divided into Pinelliae Rhizoma alkaloids(PTAs) and Pinelliae Rhizoma non-alkaloids(PTNAs) by 732 cation-exchange resin. Further in vitro investigation confirmed the significant anti-inflammatory effect of PTNAs, while such effect of PTAs was not manifest. The MS analysis showed 172 peptides and 7 organic acids in PTNAs. The peptide content in PTNAs was 63.5% measured by quantitative analysis of BCA assay, and the organic acid content was 9.92% by potentiometric titration method. The findings of this study suggested that Pinelliae Rhizoma Decoction could antagonize airway inflammation in mice by inhibiting phosphorylation of p38 and IκB and blocking the activation of MAPK and NF-κB signaling pathways, and the effective components were related to the peptides and organic acids in PTNAs. The above results lay a foundation for the research on the mechanism and material basis of Pinelliae Rhizoma in antagonizing airway inflammation.

Knockdown of long non-coding RNA HOTAIR reverses cisplatin resistance of ovarian cancer cells through inhibiting miR-138-5p-regulated EZH2 and SIRT1

BACKGROUND: Cisplatin resistance (DDP-resistance) remains one of the major causes of poor prognosis in females with ovarian cancer. Long non-coding RNAs (lncRNAs) have been shown to participate in the regulation of cellular processes, including chemoresistance. The aim of this study was to explore the role of HOX transcript antisense RNA (HOTAIR) in DDP-resistant ovarian cancer cells. METHODS: DDP-resistant ovarian cancer cell lines (SKOV3/DDP and A2780/DDP) were established. Real-time PCR, western blot, dual-luciferase reporter assay, and flow cytometry were then used to evaluate the effect of HOTAIR/miR-138-5p axis on chemoresistance of DDP-resistant ovarian cancer cells to DDP. RESULTS: We found that HOTAIR was upregulated in DDP-resistant cells, while miR-138-5p was downregulated. Knockdown of HOTAIR increased the expression of miR-138-5p in DDP-resistant cells and miR-138-5p is directly bound to HOTAIR. Upregulation of miR-138-5p induced by HOTAIR siRNA or by its mimics enhanced the chemosensitivity of DDP-resistant cells and decreased the expression of EZH2 (enhancer of zeste 2 polycomb repressive complex 2 subunit) and SIRT1 (sirtuin 1). Furthermore, the HOTAIR silencing-induced chemosensitivity of DDP-resistant cells was weakened by miR-138-5p inhibitor. CONCLUSIONS: These data demonstrate that HOTAIR acts as a sponge of miR-138-5p to prevent its binding to EZH2 and SIRT1, thereby promoting DDP-resistance of ovarian cancer cells. Our work will shed light on the development of therapeutic strategies for ovarian cancer treatment.

The effect of environment on the serum concentrations of adrenocorticotropic hormone, cortisol and testosterone / 中华物理医学与康复杂志

Objective:To investigate any effect of different environments on the levels of adrenocorticotropic hormone (ACTH), cortisol and testosterone using SAMP8 senescence-accelerated mice.Methods:Thirty SAMP8 mice were randomly divided into an enriched environment (EE) group, an impoverished environment (IE) group and a standard environment (SE) group, each of 10. The serum levels of ACTH, cortisol and testosterone were measured in all of the mice after they had lived in their respective environments for 8 weeks.Results:The average ACTH concentrations of the IE, EE and SE groups were (60.54±16.22), (48.98±15.30) and (28.49±8.24)pg/ml respectively. The average cortisol concentrations were (5.37±0.81), (4.09±0.92) and (3.19±0.88)ng/ml. The average testosterone concentrations being (2.35±0.90), (7.07±1.57) and (3.16±1.10)ng/ml. The average ACTH and cortisol levels were significantly different between the SE and IE groups. The average ACTH and testosterone levels differed significantly between the SE and EE groups and between the EE and IE groups.Conclusions:An impoverished environment can increase the secretion of ACTH and cortisol and activate the hypothalamic-pituitary-adrenal axis, at least in SAMP8 mice. An enriched environment can promote the secretion of ACTH and testosterone, but not that of cortisol.

Association between mandibular prognathism and the candidate gene GHR in the Han Chinese in Hong Kong / 医学研究生学报

Objective　Studies on the location of the susceptibility genes of mandibular prognathism are mostly conducted by association analysis. The aim of this study was to investigate the correlation between the single nucleotide polymorphism (SNP) of the candidate gene of the growth hormone receptor (GHR) and mandibular prognathism in the Han Chinese in Hong Kong. Methods We selected 7 tag SNPs (tSNP) in the GHR gene from 211 cases of mandibular prognathism and 224 controls among the Han Chinese in Hong Kong. We genotyped the tSNPs using the MassARRAY System and analyzed the association of individual SNPs and the relevant haplotypes with mandibular prognathism.　Results　The SNP rs6898743 of the GHR gene was found associated with mandibular prognathism in genotypic frequency (P = 0.036) and allelic distribution (P = 0.015), and the G allele of rs6898743 associated with a significantly decreased risk of mandibular prognathism (OR: 0.72; 95% CI: 0.55-0.94). Linkage disequilibrium analysis revealed the association of mandibular prognathism with the haplotypes GAA (P = 0.014) and GAG (P = 0.012).　Conclusion　The SNP locus rs6898743 of the GHR gene is associated with mandibular prognathism in the Han Chinese in Hong Kong

The antidiabetic effect of jejunal exclusion surgery for T2DM rats / 中华内分泌外科杂志

Objective To study the curative effects of jejunal exclusion surgery for STZ-induced T2DM SD rats.Methods 60 SD rats were induced to be the T2DM SD rats by intraperitoneal injection of streptozotocini.As a result,55 T2DM SD rats were successfully acquired which were randomly divided into 3 groups,20 rats in the jejunal exclusion group (A),20 rats in the sham operation group (B) and 15 rats in the control group (C).Jejunal exclusion surgery was performed in group A,jejunojejunostomy was performed in group B,and group C were fed normally.The body weight,fasting blood glucose,fasting plasma insuhn level and GLP-1 level were measured before operation and at the 1st,2rid,4th,8th and 16th week after operation.Results As compared with that before operation and that of the control group,the body weight in group A markedly declined at the 2nd,4th,8th and 16th week (352.14±9.00,342.84±8.90,336.64±10.26,330.34±9.12,P＜0.05).The fasting plasma glucose levels in group A markedly declined at the 2nd,4th,8th and 16th week (14.62±1.10,12.12±1.38,8.75± 1.06,7.55±1.00,P＜0.05).The fasting plasma insulin level in group A was maikedly increased at the 2nd,4th,8th and 16th week (14.62±3.10,16.12±3.38,17.75±4.06,17.55±3.10,P＜0.05).GLP-1 level in group A was markedly increased at the 1st,2nd,4th,8th and 16th week (11.02±0.85,14.42±1.18,16.02±1.59,17.62±1.02,18.12±0.71,P＜0.05).Conclusions The jejunal exclusion surgery is effective in controlling blood glucose,which is an ideal and lasting method.This surgery has also showed influence on body weight.