Advances in analytical methods for endogenous bile acids based on UPLC-MS/MS technology / 药学学报

Bile acids (BAs) are a group of endogenous steroid molecules that regulate lipid, glucose and energy metabolism. They play an important role in maintaining body homeostasis and physiological functions as key signaling molecules for host and gut microbial metabolism. The accurate characterization and quantification of BAs in vivo is of great importance in basic and clinical research. Over the past decades, enzymatic assay, enzyme-linked immunoassay, nuclear magnetic resonance (NMR), chromatography, and other related techniques have been developed and applied to the detection of BAs. The diverse structures of BAs, the existence of isomers and the complex matrix of biological samples pose great challenges for the detection of endogenous BAs. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) is a robust analytical technique that combines the rapid separation capacities of UPLC with the powerful structural identification capabilities of MS/MS, facilitating the more rapid separation, characterization and accurate quantitative of target analytes in biological samples. UPLC-MS/MS has been widely used in the quantitative analysis of BAs in recent years for its high selectivity, high sensitivity, and high accuracy. This paper summarized the biosynthetic pathways of BAs, sample pretreatment methods, common analytical detection techniques, and highlights the current status of the application of UPLC-MS/MS technology in the analysis of endogenous BAs over the past five years, to provide a reference for the accurate detection of endogenous BAs and further research development and application.

Mechanism of seahorse ameliorating depression-like behavioral and pathological changes caused by chronic stress in zebrafish / 中国药理学通报

Aim To investigate the effect of marine herbal seahorse on chronic unpredictable mild stress ( CUMS ) -induced depression-like model in zebrafish. Methods Adult zebrafish were divided into control, Stress,Stress + low dose (Stress +0.044% SH) and Stress + high dose (Stress +0. 22% SH) seahorse intervention groups, and depression-like behavior was identified by novel tank test (NTT), cortisol, interleukin ( IL )-6 and interferon (IFN )-γ levels were detected by ELISA. The levels of dopamine (DA) ,norepinephrine (NE), 5-hydroxytryptamine (5-HT) and 5-hydroxyin-doleacetic acid (5-HIAA) were determined by high performance liquid chromatography. The mRNA expression levels of tryptophan hydroxylase(TPH)-2 and 5-HT2A receptor were measured by real-time quantitative PCR. Results Compared with the control group, the Stress group showed significantly longer latency to reach the top in NTT, significantly reduced number of transfers to the top region and top residence time, significantly increased levels of cortisol and IL-6, IFN-γ protein, significantly reduced levels of DA and 5-HT in brain as well as increased metabolism rate of 5-HT, while 5-HT2A mRNA expression was up-regulated and TPH2 mRNA expression was down-regulated. In contrast, low-dose seahorse intervention effectively reduced anxiety, decreased cortisol and IL-6 and IFN-γ concentrations, increased monoamine neurotransmitter levels and reversed dysregulation of the 5-HT ergic system in CUMS zebrafish. Conclusion Seahorse may exert an-tidepressant effects through anti-inflammation and mod¬ulation of monoamine neurotransmitter levels.

Quality evaluation of Compound Cheqian Tablets based on UPLC-Q-TOF-MS/MS, network pharmacology and "double external standards" QAMS / 中国中药杂志

The Compound Cheqian Tablets are derived from Cheqian Power in Comprehensive Recording of Divine Assistance, and they are made by modern technology with the combination of Plantago asiatica and Coptis chinensis. To investigate the material basis of Compound Cheqian Tablets in the treatment of diabetic nephropathy, in this study, the chemical components of Compound Cheqian Tablets were characterized and analyzed by UPLC-Q-TOF-MS/MS, and a total of 48 chemical components were identified. The identified chemical compounds were analyzed by network pharmacology. By validating with previous literature, six bioactive compounds including acteoside, isoacteoside, coptisine, magnoflorine, palmatine, and berberine were confirmed as the index components for qua-lity evaluation. Furthermore, the content of the six components in the Compound Cheqian Tablets was determined by the "double external standards" quantitative analysis of multi-components by single marker(QAMS), and the relative correction factor of isoacteoside was calculated as 1.118 by using acteoside as the control; the relative correction factors of magnoflorine, palmatine, and berberine were calculated as 0.729, 1.065, and 1.126, respectively, by using coptisine as the control, indicating that the established method had excellent stability under different conditions. The results obtained by the "double external standards" QAMS approximated those obtained by the external standard method. This study qualitatively characterized the chemical components in the Compound Cheqian Tablets by applying UPLC-Q-TOF-MS/MS and screened the pharmacodynamic substance basis for the treatment of diabetic nephropathy via network pharmacology, and primary pharmacodynamic substance groups were quantitatively analyzed by the "double external stan-dards" QAMS method, which provided a scientific basis for clarifying the pharmacodynamic substance basis and quality control of Compound Cheqian Tablets.

Research progress in the pathogenesis mechanism of dental fluorosis / 中华口腔医学杂志

Dental fluorosis is a developmental disturbance of dental enamel caused by excessive fluoride intake during tooth development, leading to the changes in morphology, structure and function of tooth enamel, which can affect the aesthetics and function of teeth. There are many factors which may account for the occurrence of dental fluorosis. However, the pathogenesis mechanism underlying dental fluorosis has not been fully clarified.In recent years, researches in the fields of fluoride-induced stress response pathways, signaling pathways and apoptosis at the molecular and genetic level had provided extensive knowledge of dental fluorosis. This article focuses on the latest research progress in the mechanism of dental fluorosis, which include the effects of fluoride on ameloblasts and enamel matrix proteins, genetic polymorphism and dietary nutrients, in order to provide new references for the targeted prevention and treatment of dental fluorosis.

Research progress in the association of peri-implant diseases and metabolic syndrome / 中华口腔医学杂志

Peri-implant disease, an important group of diseases that cause implant failure, are associated with metabolic abnormality. Metabolic syndrome (MetS) is a common metabolic disorder comprising abdominal obesity, hyperglycemia, systemic hypertension and atherogenic dyslipidemia. Previous studies had reported that MetS and its diversified clinical manifestations might be associated with peri-implant diseases, but the relationship and underlying mechanisms were unclear. This review aims to explore the relationship between MetS and peri-implant disease, in order to provide beneficial reference for the prevention and treatment of peri-implant disease in patients with MetS.

Impact of nonsteroidal anti-inflammatory drugs on efficacy of anti-PD-1 therapy for primary liver cancer / 南方医科大学学报

OBJECTIVE@#To assess the impact of nonsteroidal anti-inflammatory drugs (NSAIDs) on clinical outcomes of patients receiving anti-PD-1 immunotherapy for hepatocellular carcinoma.@*METHODS@#We conducted a retrospective study among 215 patients with primary liver cancer receiving immunotherapy between June, 2018 and October, 2020. The patients with balanced baseline characteristics were selected based on propensity matching scores, and among them 33 patients who used NSAIDs were matched at the ratio of 1∶3 with 78 patients who did not use NSAIDs. We compared the overall survival (OS), progression-free survival (PFS), and disease control rate (DCR) between the two groups.@*RESULTS@#There was no significant difference in OS between the patients using NSAIDs (29.7%) and those who did not use NSAIDs (70.2%). Univariate and multivariate analyses did not show an a correlation of NSAIDs use with DCR (univariate analysis: OR=0.602, 95% CI: 0.299-1.213, P=0.156; multivariate analysis: OR=0.693, 95% CI: 0.330-1.458, P=0.334), PFS (univariate analysis: HR=1.230, 95% CI: 0.789-1.916, P=0.361; multivariate analysis: HR=1.151, 95% CI: 0.732-1.810, P=9.544), or OS (univariate analysis: HR=0.552, 95% CI: 0.208-1.463, P=0.232; multivariate analysis: HR=1.085, 95% CI: 0.685-1.717, P=0.729).@*CONCLUSION@#Our results show no favorable effect of NSAIDs on the efficacy of immunotherapy in patients with advanced primary liver cancer, but this finding still needs to be verified by future prospective studies of large cohorts.

Investigation of Effect of Polyethylene Glycol 400 on Pharmacokinetics and Anti-inflammatory Effect of Baicalin Based on UPLC-MS/MS and Molecular Docking Techniques / 中国实验方剂学杂志

Objective:To investigate the effects of polyethylene glycol 400 （PEG400） on the pharmacokinetics and anti-inflammatory effect of baicalin， and to preliminarily explore the anti-inflammatory effects of baicalin and its main metabolite baicalein 6-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucuronide （B6G） by molecular docking. Method:Rats were randomly divided into two groups with water and PEG400 as the dissolving matrix， and rats were administrated the equal dose of baicalin aqueous solution （baicalin+water group） and baicalin PEG400 solution （baicalin+PEG400 group）. After the plasma samples were processed at different time periods， the concentrations of baicalin and B6G in rat plasma were determined by ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and pharmacokinetic parameters were processed by DAS 3.2.2 software. Mice were randomly divided into a blank group （normal saline， 20 mL·kg^-1）， aspirin group （dose of 0.2 g·kg^-1）， baicalin/baicalin+PEG400 high and low dose （3.0， 1.5 g·kg^-1） groups， after continuous administration for 7 days， the mouse ear swelling and foot swelling models were established， and the swelling degree and swelling inhibition rate were calculated. Result:The pharmacokinetic study showed that compared with baicalin+water group， the plasma concentrations of baicalin and B6G increased after administration of baicalin PEG400 solution， and the area under the curve （AUC_0-<italic>_t</italic>） increased by 2.36， 1.97 times， and the peak concentration （<italic>C</italic>_max） increased by 2.12， 1.65 times， respectively. The results of mouse ear and foot swelling inflammation models showed that the anti-inflammatory effect was enhanced after intragastric administration of baicalin PEG400 solution. In addition， molecular docking results showed that baicalin and B6G could site bind to multiple target proteins ［tumor necrosis factor （TNF）-<italic>α</italic>， interleukin （IL）-6， IL-1<italic>β</italic>， prostaglandin E₂ （PGE₂） and nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）］ with higher affinity， which was superior to the positive drug aspirin. Conclusion:PEG400 can increase the plasma concentration of baicalin and its main metabolite B6G， and enhance the anti-inflammatory effect. Baicalin and B6G can form strong hydrogen bonds with various inflammatory factors and of nuclear transcription factors， it is speculated that baicalin and B6G jointly play an anti-inflammatory role.

Effect of molecular weight of polyethylene glycol on pharmacokinetics of baicalin / 药学学报

The aim of this study was to investigate the effects of polyethylene glycol (PEGs) with different molecular weights (MW: 400, 1 000, 4 000) on the pharmacokinetics of baicalin, and preliminarily analyze its mechanism. Rats were gavaged with baicalin (168 mg·kg-1) + aqueous solution or baicalin + PEGs solution and plasma samples were collected from 0 to 24 h after administration. The concentration of baicalin and its main metabolite baicalein 6-O-β-D-glucuronide (B6G) were determined at different time points by UPLC-MS/MS, and the pharmacokinetic parameters were calculated with DAS 3.0 software. The results showed that PEGs with different molecular weights could effectively increase the AUC0-t of baicalin and B6G, increase the Cmax, and prolong the t1/2, effectively increasing the concentration of baicalin and B6G in vivo. The mechanism may be by promoting the activity of uridine diphosphate glucuronosyl-transferases 1A8 (UGT1A8) and 1A9 (UGT1A9), thereby increasing the transformation rate of baicalin and B6G. The rate of metabolism of B6G was faster than that of baicalin, suggesting that PEGs had a higher affinity for UGT1A8, and PEG400 had the most significant effect. The purpose of this study was to provide a basis for the clinical safe use of baicalin and other flavonoids and the design of new dosage forms with the participation of PEGs. The animal experiment protocol in this study was approved by the Experimental Animal Ethics Committee of Guizhou Medical University.

Simultaneous determination of multiple bioactive constituents in Abelmoschi Corolla by UFLC-QTRAP-MS/MS / 中国中药杂志

A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 38 active components in Abelmoschi Corolla, including flavonoids, organic acids, nucleosides and amino acids, so as to investigate the effects of different harvesting and processing methods on multi-active components in Abelmoschi Corolla. The chromatographic separation was performed on a XBridg®C_(18) column(4.6 mm×100 mm, 3.5 μm) with(0.1% formic acid water) methanol-acetonitrile(1∶1) as the mobile phase for gradient elution at 30 ℃. The flow rate was 0.5 mL·min~(-1). The components were detected in a multiple-reaction monitoring(MRM) mode. The gray relational analysis(GRA) was used to comprehensively evaluate the multiple active components of Abelmoschi Corolla at different harvesting times and drying temperatures. The results showed that 38 components had a good linearity with correlation coefficients all above 0.999 0. The method featured a good precision, repeatability and stability with the relative stan-dard deviations(RSDs) of less than 5.0%. Recoveries ranged from 98.06% to 104.4% with RSD between 0.22% and 4.9%. The results of GRA indicated that a better quality in the samples collected on September 9 th. Samples dried at 90 ℃ had a better quality. The established method is accurate and reliable, and can be used to assess the internal quality of Abelmoschi Corolla. This study can provide basic materials for determining appropriate harvesting time and processing method of Abelmoschi Corolla.

Research progress in molecular biology of Lonicerae Japonicae Flos / 中国中药杂志

Molecular biology is a new subject that clarifies the phenomena and nature of life at the molecular level. Its development provides new biotechnology and methods for the study of traditional pharmacognosy. The formation of molecular biology has brought the development of pharmacognosy into a new era of gene research. Lonicerae Japonicae Flos is a classical Chinese medicine. Many scholars of home and abroad have carried out relevant studies on its molecular biology on the basis of the in-depth study with traditional methods, and have achieved certain results. In order to provide references on the method, technical for promoting the modernization of Lonicerae Japonicae Flos, and the development, protection, and utilization of other traditional Chinese medicine resources. This article summarized the application status of molecular biology methods and techniques on the identification, biosynthesis of active constituents, and molecular mechanism of secondary metabolite under stress conditions of Lonicerae Japonicae Flos in recent years. In hybridization technology of tag(RFLP), molecular markers based on PCR(RAPD, AFLP, SSR and ISSR), based on DNA sequence analysis of SNP and DNA barcode for the variety identification, diagnosis, identification of Lonicerae Japonicae Flos, and so forth in detail. At the same time, it is proposed that multi-omics technology can be used to build systems biology technology and platforms, and establish related models of secondary metabolite biosynthesis, so as to deepen acknowledge the molecular mechanism of the active component biosynthesis of Lonicerae Japonicae Flos and the accumulation of metabolites, life activities of other medicinal plants under adverse environment, then to regulate them.

Analysis and evaluation of dynamic accumulation of multiple bioactive constituents in Spatholobi Caulis / 中国中药杂志

A method was established for simultaneous determination of 21 active constituents including flavanols, isoflavones, flavonols, dihydroflavones, dihydroflavonols, chalcones, pterocarpan, anthocyanidins and phenolic acids in Spatholobi Caulis by ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS). Then, it was employed to analyze and evaluate the dynamic accumulation of multiple bioactive constituents in Spatholobi Caulis. The chromatographic separation was performed on a XBridge®C_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.3% formic acid aqueous solution-methanol, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. A comprehensive evaluation of the multiple bioactive constituents was carried out by gray correlation analysis(GRA). The 21 target components showed good linearity(r>0.999 0) in the range of the tested concentrations. The average recovery rates of the 21 components were from 97.46% to 103.6% with relative standard deviations less than 5.0%. There were differences in the contents of 21 components in Spatholobi Caulis at diffe-rent harvest periods. Spatholobi Caulis had high quality from early November to early December, which is consistent with the local tradi-tional harvest period. This study reveals the rule of the dynamic accumulation of 21 components in Spatholobi Caulis and provides basic information for the suitable harvest time. At the same time, it provides a new method reference for the comprehensive evaluation of the internal quality of Spatholobi Caulis.

Stimultaneous determination of multiple bioactive constituents in Panacis Japonici Rhizoma processed by different methods and grey relational analysis / 中国中药杂志

A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC₁₈(4.6 mm×100 mm, 3.5 μm) at 30 °C with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min⁻¹, using multiple-reaction monitoring (MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.

Clinical study of cancer-related fatigue in patients with advanced gastric cancer / 肿瘤研究与临床

Objective To study the relationship between cancer-related fatigue (CRF) and clinicopathological features and quality of life in patients with advanced gastric cancer. Methods The clinical and pathological data of 182 patients with advanced gastric cancer were collected, The CRF level and quality of life were assessed by the brief fatigue inventory (BFI), European Organization for Research on Treatment of Cancer (EORTC) life quality scale QLQ-C30 (V3.0). The correlations between CRF and clinicopathological features and quality of life were analyzed. Results One hundred twenty-two patients with advanced gastric cancer were associated with different degrees of fatigue. The CEF BFI score had correlation with the age(F=6.637,P=0.001),PS score(F=15.559,P=0.000),the World Health Organization(WHO) pathologic type (F=7.352, P=0.000). The CRF index of the 50-59 age group was higher than that of other age groups; the CRF BFI scores of PS score 1 and 2 were higher than that of 0 group; CRF BFI score of gastric signet ring cell carcinoma was significantly higher than that of gastric squamous cell carcinoma. The Pearson correlation analysis showed that the CRF BFI score was positively correlated with mood (r=0.423, P=0.008). There were significant differences in body function and mood (F=-16.254, F=-13.256, both P<0.05)and fatigue,pain and loss of appetite among 3 CRF groups(F=12.075,F=14.361,F=13.837, all P<0.05). There was a significant difference in the mean CRF BFI score between the three groups before and after chemotherapy(tSD=6.981,tPR=6.903,tPD=7.437,all P<0.05). Compared with pre-chemotherapy, the mean CRF BFI score of SD and PR group decreased after 3 months follow-up, score of PD group increased, the differences were statistically significant(t=2.371,t=6.912,t=18.237,all P<0.05).Conclusions The incidence of CRF in patients with advanced gastric cancer is very high,the degree of CRF has correlation with age, PS score and pathological type. Relieve symptoms such as fatigue, pain and loss of appetite and control adverse reactions could help to reduce the degree of CRF.

Clinical study of cancer-related fatigue in patients with advanced gastric cancer / 肿瘤研究与临床

Objective To study the relationship between cancer-related fatigue (CRF) and clinicopathological features and quality of life in patients with advanced gastric cancer. Methods The clinical and pathological data of 182 patients with advanced gastric cancer were collected, The CRF level and quality of life were assessed by the brief fatigue inventory (BFI), European Organization for Research on Treatment of Cancer (EORTC) life quality scale QLQ-C30 (V3.0). The correlations between CRF and clinicopathological features and quality of life were analyzed. Results One hundred twenty-two patients with advanced gastric cancer were associated with different degrees of fatigue. The CEF BFI score had correlation with the age(F=6.637,P=0.001),PS score(F=15.559,P=0.000),the World Health Organization(WHO) pathologic type (F=7.352, P=0.000). The CRF index of the 50-59 age group was higher than that of other age groups; the CRF BFI scores of PS score 1 and 2 were higher than that of 0 group; CRF BFI score of gastric signet ring cell carcinoma was significantly higher than that of gastric squamous cell carcinoma. The Pearson correlation analysis showed that the CRF BFI score was positively correlated with mood (r=0.423, P=0.008). There were significant differences in body function and mood (F=-16.254, F=-13.256, both P<0.05)and fatigue,pain and loss of appetite among 3 CRF groups(F=12.075,F=14.361,F=13.837, all P<0.05). There was a significant difference in the mean CRF BFI score between the three groups before and after chemotherapy(tSD=6.981,tPR=6.903,tPD=7.437,all P<0.05). Compared with pre-chemotherapy, the mean CRF BFI score of SD and PR group decreased after 3 months follow-up, score of PD group increased, the differences were statistically significant(t=2.371,t=6.912,t=18.237,all P<0.05).Conclusions The incidence of CRF in patients with advanced gastric cancer is very high,the degree of CRF has correlation with age, PS score and pathological type. Relieve symptoms such as fatigue, pain and loss of appetite and control adverse reactions could help to reduce the degree of CRF.

Comparison on sensitivity and specificity of serum miR-122 as biomarkers of drug-induced liver injury / 中国药理学与毒理学杂志

OBJECTIVE To make a comparison of sensitivity and specificity between serum miR-122 and traditional biomarkers of drug-induced liver injury. METHODS Acetaminophen (APAP, 1250 mg · kg-1, ig), α-naphthylisothiocyanate (ANIT, 150 mg · kg-1, ig), methionine-choline deficient diet (MCDD, free feeding) and carbon tetrachloride (CCl4, 50%, ip) were used to establish hepatocellular injury, cholestasis, steatosis and fibrosis models, respectively, which were used to evaluate the sensitivity of serum miR-122 as a biomarker of drug-induced liver injury, when compared with the traditional biomarkers. Isoprenaline hydrochloride (IH) and gentamicin (GM) were used to establish myocardial and renal injury models, respectively, which were used to evaluate the specificity of serum miR-122, when compared with the tradi-tional biomarkers. Serum and liver tissues were collected at different time points during the study. Tradi-tional biomarkers such as glutamic-pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT) and total bilirubin (TBIL) were measured with an automatic biochemistry analyzer. MiR-122 was detected with real- time quantitative PCR. Histopathological examination with HE staining was performed for liver tissues. RESULTS Serum miR-122 increased significantly earlier [miR-122 was signifi-cantly increased (>2 fold) at 1.5 h, 3 h, 2 weeks and 4 weeks after treatment in the four models respec-tively, while no significant increase (<2 fold) was observed for GPT, GOT and TBIL at 6 h, 12 h, 3 weeks and 6 weeks after treatment in the four models respectively] and more signficantly (the maximum fold change for miR-122 was 235.8, 202.7, 73.8 and 600.3 for the four models, respectively. For the GPT, the maximum fold change was 9.5, 3.9, 2.5 and 6.6, respectively; 6.0, 2.4, 1.4 and 3.5 respectively for GOT; 2.6, 2.3, 1.2 and 1.8 respectively for TBIL) than that of traditional biomarkers in hepatocellular injury, cholestasis, steatosis and fibrosis models, when compared with the control group. In the myocar-dial injury model, a significant increase of GOT was observed after IH treatment (2.1 fold), while no change was observed for serum miR-122. In the renal injury model, no false positive results were observed for serum miR-122. CONCLUSION Serum miR-122 can be used as a biomarker of drug-induced liver injury and serum miR-122 is more sensitive and specific than traditional biomarkers (such as GPT, GOT and TBIL).

Study of dose variations induced by MLC weight effect for IMRT delivery / 医疗卫生装备

Objective To evaluate dose variations induced by gravity of multi-leaf collimator to provide references for clinical intensity-modulated radiotherapy.Methods Two-dimensional dose distributions in the central plane of IMRT fields were measured by use of a 2D ion chamber array.All measurements were repeated at two collimator angles (C=0 and 90°),for each of the following gantry angles:G=0 and 270°.Comparisons were made to dose distributions generated at G=0°and their differences were analyzed using gamma index analysis (3％/3 mm and 1％/1 mm).Results Under the radiation field of 10 cm×10 cm,the gamma passing rate was higher than 99％ for 3％ 3 mm anch close to 95％ for 1％/1mm Under a 3％/3mm error standard,the average matching rate for step & shoot fields was (96.46±0.33)％ and for DMLC fields was (94.67±0.54)％ at C=0°;The average matching rate for step & shoot fields was (94.59±0.47)％ and for DMLC fields was (92.60±0.52)％ at C=90°.Under a 1％/1mm error standard,the average matching rate for step & shoot fields was (89.83 ±1.06)％ and for DMLC fields was (85.84±0.57)％ at C=0°;The average matching rate for step & shoot fields was (86.91 ±1.71)％ and for DMLC fields was (83.89±0.69)％ at C=90°.Concusion MLC weight effect affects IMRT delivery dose,and DMLC fields are more sensitive to gravity than step & shoot fields.

Construction and in vitro study of eukaryotic expression vector carrying GITRL and IL-21 gene / 中国实验血液学杂志

The aim of this study was to construct the eukaryotic expression vector carrying glucocorticoid-induced tumor necrosis factor receptor ligand (GITRL) and interleukin-21 (IL-21) gene for transfection into chronic myeloid leukemia (CML) dervied dendritic cell (DC), so as to provide an effective platform for exploring the function of target gene in CML. The recombinant eukaryotic expression vector was transfected to the purified DC by Liposome-mediated method, the interleukin-2 (IL-2) and Interferon-γ (IFN-γ) expression of transfected DC were analyzed by ELISA. Further, the transfected DC with purified NK were mixed and cultured to be DC-CIK, the lactate dehydrogenase release assay was performed to measure the killing activity of DC-CIK. The results indicated that the sequence of cloned target gene was same as that in GenBank. The size of endonuclease products by restriction enzyme were same as the predict one. The concentration of Interleukin-2 (IL-2) and interferon-γ (IFN-γ) in transfected DC all increased. The NK kill activity became stronger while induced by transfected DC. It is concluded that DC transfected by IL-21 and GITRL gene has the ability of self-activation, up-regulate cytokine secretion. Further, the results would be help to provide the theoretical evidence of advanced immunotherapy for treatment of CML patients who showed no reaction to tyrosine kinase inhibitor.

Surveillance and early diagnosis of small hepatocellular carcinoma with contrast-enhanced ultrasonography / 中国癌症杂志

Background and purpose:The incidence of liver cancer is high in China. Primary liver cancers usually occur in patients with liver cirrhosis, which is a challenge for the early diagnosis of liver cancer. Our purpose is to investigate the efifcacy of contrast-enhanced ultrasonography (CEUS) in the early identiifcation and diagnosis of small hepatocellular carcinoma (HCC) by regularly tracking and supervising the high risk population. Methods:A total of 320 high risk patients of HCC admitted in our hospital from February 2011 to November 2013 were enrolled in this prospective study. All patients underwent conventional ultrasound and hepatic CEUS. The differential diagnosis of malignant HCCs from benign ones was based on the enhancement patterns of hepatic lesions in different phases on CEUS. Results:Twenty patients were diagnosed as small HCC among 320 HCC high risk patients who were under regular surveillance using CEUS and all were pathologically conifrmed. Seven of the 20 HCC cases were smaller than 1.0 cm and 13 measured 1.1-2.0 cm. There were 6 (30.0%) HCCs presented as“early wash-in and slow wash-out”atypical pattern of HCC. The small size of the lesion and iso-echogenicity were the main factors of atypical pattern of HCC on CEUS.Conclusion:Ultrasonography and CEUS surveillance is a useful strategy for the early detection of small HCCs in high risk patients, which can help them to receive proper therapeutic management in time.

Monitoring the supporting role of traditional Chinese medicine intervention in the treatment of high-risk status of liver cancer by color doppler ultrasound / 国际中医中药杂志

Objective To study the supporting role of traditional Chinese medicine GanFuAn in the treatment of high-risk state of liver cancer by color Doppler ultrasound.Methods Based on a number of joint detection of tumor markers,85 patients of liver cancer were selected from patients with chronic hepatitis B.Among these 85patients,50 patients were selected in a treatment group and treated with Chinese herbal preparation GanFuAn,with ultrasound and tumor markers serving as joint monitoring; while the other 35 patients were selected in a control group and treated with conventional treatment.Results In the treatment group,the liver tumor markers or liver tumor continually diminished.5 cases showed tumor disappear.38 cases showed effective results,while 7 cases showed no effects.The total incidence rate of liver cancer was 14％;while in the control group,none case showed tumor disappear.20 cases showed effects and 15 cases showed none effects.The cumulative incidence rate of liver cancer induced by chronic hepatitis was 43％,with all the cases have been confirmed by pathology.Conclusion Color doppler ultrasound was an intuitive,non-invasive dynamic method to monitor liver tumor's growth and decline.

Specific targeting of angiogenesis in lung cancer with RGD-conjugated ultrasmall superparamagnetic iron oxide particles using a 4.7T magnetic resonance scanner / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Angiogenesis is an essential step for tumor development and metastasis. The cell adhesion molecule avβ3 integrin plays an important role in angiogenesis and is a specific marker of tumor angiogenesis. A novel avβ3 integrin- targeted magnetic resonance (MR) imaging contrast agent utilizing Arg-Gly-Asp (RGD) and ultrasmall superparamagnetic iron oxide particles (USPIO) (referred to as RGD-USPIO) was designed and its uptake by endothelial cells was assessed both in vitro and in vivo to evaluate the angiogenic profile of lung cancer.</p><p><b>METHODS</b>USPIO were coated with -NH3+ and conjugated with RGD peptides. Prussian blue staining was performed to evaluate the specific uptake of RGD-USPIO by human umbilical vein endothelial cells (HUVECs). Targeted uptake and subcellular localization of RGD-USPIO in HUVECs were confirmed by transmission electron microscopy (TEM). The ability of RGD-USPIO to noninvasively assess avβ3 integrin positive vessels in lung adenocarcinoma A549 tumor xenografts was evaluated with a 4.7T MR scanner. Immunohistochemistry was used to detect avβ3 integrin expression and vessel distribution in A549 tumor xenografts.</p><p><b>RESULTS</b>HUVECs internalized RGD-USPIO significantly more than plain USPIO. The uptake of RGD-USPIO by HUVECs could be competitively inhibited by addition of free RGD. A significant decrease in T2 signal intensity (SI) was observed at the periphery of A549 tumor xenografts at 30 minutes (P < 0.05) and 2 hours (P < 0.01) after RGD-USPIO was injected via the tail vein. Angiogenic blood vessels were mainly distributed in the periphery of tumor xenografts with positive avβ3 integrin expression.</p><p><b>CONCLUSIONS</b>RGD-USPIO could specifically label avβ3 integrin and be taken up by HUVECs. This molecular MR imaging contrast agent can specifically evaluate the angiogenic profile of lung cancer using a 4.7T MR scanner.</p>