1.Development and verification of high performance liquidchromatography for determination of ethylenediaminetetraacetic acid disodium salt residue in NMM tumor DNA vaccine
Chinese Journal of Biologicals 2023;36(7):855-861
Objective To develop and verify a high performance liquid chromatography(HPLC) method for the determination of ethylenediaminetetraacetic acid disodium salt(EDTA-2Na) residues in the bulk of NMM tumor DNA vaccine for the quality control of DNA vaccine.Methods After NMM tumor DNA vaccine bulk was complexed with copper sulfate,a HPLC method for the determination of EDTA-2Na residues was developed with Agilent ZORBA XSB-C18(150 mm × 4.6 mm,5 μm) as the chromatographic column,water,tetrabutylammonium hydroxide 10% and acetonitrile solution(74.5:0.5:25)as the mobile phase.The detection method was as follows:the detection wavelength was 254 nm,the flow rate was 0.8 mL/min,the column temperature was 20 ℃ and the injection volume was 20 μL.The method was verified for the specificity,linear range,limit of detection(LOD),limit of quantification(LOQ),solution stability,durability,accuracy and precision,and used to detect the EDTA-2Na residues in several batches of DNA vaccine bulk.Results When EDTA-2Na control and DNA vaccine bulk with EDTA-2Na reacted with copper sulfate,the absorption peak appeared at around 5.3 min,while no absorption peak was observed when DNA vaccine bulk reacted with copper sulfate;In the range of 4~400 μg/mL,the control solution concentration showed a good linear relationship with the peak area,R~2=0.999 9;The LOD of the method was 10 ng/mL,and the LOQ was 40 ng/mL;The solution of control and sample was stable after placed for 12 h;When the detection conditions changed slightly(different mobile phase ratio,flow rate and column temperature),the influence on the detection results was within acceptable range;The average recovery rate of EDTA-2Na in low,medium and high concentration standard added samples was 101.38% with the RSD of 0.39%;0.1 mg/mL control solution was injected continuously for 6 times,and the peak area RSD was 0.04%.EDTA-2Na was not detected in 6 sample solution,and the peak area RSD of DNA vaccine bulk with EDTA-2Na solution was 0.02%,indicating a good intermediate precision.EDTA-2Na residue was not detected in these batches of DNA vaccine bulk.Conclusion The developed method is simple,accurate,reliable with good specificity,which can be used for the determi-nation of EDTA-2Na residues in DNA vaccine bulk.
2.Mechanism of Xiao Yao San influences concentration of Ca2+ intracellular in cultured rat hippocampal neurons in the state of chronic stress
Haiqing AO ; Qi SUN ; Zhiwei XU ; Wenzhu WANG ; Wenjun FU
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(9):820-822
Objective To investigate the effects of Xiao Yao San on intracellular Ca2 + concentration in cultured rat hippocampal neurons in the state of chronic stress and study the mechanism of chronic stress injuring and XiaoYao San protecting. Methods MK-801 acts as tool,cultured rat hippocampal neurons were divided into seven groups, those were group 1 (control), group 2 (normal serum), group 3 (normal serum + Glu), group 4 (model serum + Glu), group 5 (model serum + Glu + MK-801), group 6 (Xiaoyaosan + Glu), group 7 (Xiaoyaosan + Glu + MK-801). to detect intracellular Ca2+ concentration in cultured hippocampal neurons in the simulated micro - environment of chronic stress and after intervention with the serum treated with Xiao Yao San by confocal laser microscope at the same period of time. Results Compared with group 1 (779.97 ± 36.81), concentration of Ca2+ intracellular of group 2 (1092.38 ± 36.41), group 3 (1472.49 ± 76. 19), group 4 (1509.52 ±104.69) and group 5 (1186.97 ±41.92) all increased significantly (P<0.01) ,group 6 (908.74 ±40.24) increased too (P < 0.05), compared with group 2, concentration of Ca2 + intracellular of group 3,4 and 5 all increased significantly (P < 0.01), but group 7 (721.99 ± 60.33) decreased significantly (P < 0.01). Compared with group 4, concentration of Ca2+ intracellular of group 6 and 7 decreased significantly (P< 0.01), group 5 decreased too (P < 0.05), compared with group 6, concentration of Ca2 + intracellular of group 5 increased significantly (P < 0.01),when group 7 decreased significantly (P<0.01). Conclusion Serum of chronic stress treated with Xiao Yao San has the effect of inhibiting Ca2+ overload in hippocampal neurons,it may work through a variety of signaling pathways including Glu-NR-Ca2+ to maintain the steady-state of Ca2+ concentration in hippocampal neurons, and then to protect neurons from the neurotoxic effects of excitatory.
3.Expression of Golgi Phosphoprotein 3 in Human Hepatocellular Carcinoma and Its Clinicopathological Significance
Chengshuo ZHANG ; Baomin CHEN ; Ao JIAO ; Ning SUN ; Jialin ZHANG
Journal of China Medical University 2017;46(3):214-218
Objective To investigate the expression characteristics of Golgi phosphoprotein 3(GOLPH3)in human hepatocellular carcinoma (HCC)and explore its clinicopathological significance. Methods The expressions of GOLPH3 protein was detected in 132 cases of paired paraf-fin embedded HCC specimens and pericarcinoma tissues using immunohistochemical staining ,and the relation of the expression of GOLPH3 to clinicopathologic features was analyzed. Meanwhile,the expression and distribution of GOLPH3 in HCC cells was observed by laser confocal mi-croscopy. Results The positive expression rates of GOLPH3 in HCC and pericarcinoma tissues were 70.0%(92/132)and 42.4%(56/132) (P<0.001),respectively. The incidence of portal vein tumor thrombus in high and low GOLPH3 expression groups of HCC were 21.2%(14/66) and 6.1%(4/66)(P<0.05),respectively. The expression rate of GOLPH3 in HCC was significantly higher than that in pericarcinoma tissues, and the expression of GOLPH3 in HCC was positively related to portal vein tumor thrombus. In addition ,GOLPH3 was mainly expressed in cyto-plasm of HCC cells,and there was also scattered distribution in the nucleus. Conclusion GOLPH3 acts as an oncogene and may play vital roles in the carcinogenesis and development of HCC.
4.The Analysis Based on Quality Function Deployment (QFD) Method of Clinical Trial Management System Requirements
Qian ZHANG ; Ruihua SUN ; Lin LI ; Yumeng WANG ; Ao HUANG
Chinese Journal of Medical Science Research Management 2014;27(3):329-332
It is imperative to apply information technology in the area of management of clinical research so as to ensure the quality of clinical trials and to improve management efficiency.In this study,the based analysis method was the quality function deployment (QFD).This methodology is used to analysis the clinical trials management information system on a hospital directly under the Ministry of Health.It ensured user participation,lay a solid foundation for software engineers on system design.
5.Cultivation,identification and sorting of bronchoalveolar stem cells derived from mouse lung
Shen QIAN ; Jianghong AN ; Xujun AO ; Jianguo SUN ; Zhengtang CHEN
Journal of Third Military Medical University 2003;0(09):-
Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P
6. Effects of recombinant human granulocyte macrophage colony stimulating factor gel on treatment of full-thickness frostbite wounds on foot and hand
Zhan′ao SUN ; Xiuhang ZHANG ; Yan XUE ; Xin ZHOU ; Xinxin CHEN ; Xinxin GAO ; Jia′ao YU
Chinese Journal of Burns 2020;36(2):117-121
Objective:
To explore the effects of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) gel on treatment of thefull-thickness frostbite wounds on foot and hand.
Methods:
From November 2013 to April 2017, a total of 45 patients of 71 full-thickness frostbite wounds on foot and hand meeting the inclusion criteria were admitted to the First Hospital of Jilin University and the prospective randomized controlled study was done. The patients were divided into rhGM-CSF group of 24 patients with 35 wounds and control group of 21 patients with 36 wounds according to the random number table. There were 20 males and 4 females, aged (38±13) years among patients in rhGM-CSF group, and there were 19 males and 2 females, aged (36±14) years among patients in control group. Patients in 2 groups were performed with the same systemic treatment of rewarming, anti-inflammation, pain relief, anti-infection, anti-coagulation, and thrombolysis. Wounds of patients in rhGM-CSF group and control group were respectively treated with rhGM-CSF gel and aloe vera gel for external usage with 10 mg for every square centimeter and dressing change once every 24 hours, until wounds healed completely. The wound inflammatory response was scored on treatment day (TD) 1, 3, 7, 14, wound secretion was collected for bacteria culture and positive bacteria detection rate was calculated before treatment and on TD 6 and 12, adverse drug reaction after drug use was observed, and the complete wound healing time was recorded. Data were processed with Fisher′s exact probability test, analysis of variance for repeated measurement,
7.The status of anti-assess on peer review of scientific research in China
Huan LI ; Ruihua SUN ; Cunxia YANG ; Zelong GU ; Yumeng WANG ; Ao HUANG
Chinese Journal of Medical Science Research Management 2015;28(1):13-15,19
Peer review anti assessment includes the evaluation of expert and index system in two parts.The current status of anti-assessment study mainly focused on expert,much of the index system anti-assessment study reports.As can be seen from the status,the anti-assessment system not yet formed and the application is rare.In urgent need of further research to improve our peer-reviewed scientific research.
8.Remnant stomach-jejunal dual pathways reconstruction after laparoscope-assisted radical proximal gastrectomy
Feng QIAN ; Bo TANG ; Yan SHI ; Yongliang ZHAO ; Huaxin LUO ; Gang SUN ; Ao MO ; Peiwu YU
Chinese Journal of Digestive Surgery 2008;7(3):174-176
Objective To investigate the value of remnant stomach-jejunal dual pathways reconstruction after laparoscope-assisted radical proximal gastrectomy in the treatment of upper gastric cancer. Methods Twenty-five patients with upper gastric cancer underwent laparoscope-assisted radical proximal gastrectomy and the remnant distal stomach was preserved for side-to-side remnant stomach-jejunal anastomosis and end-to-side jejuno-jejunal anastomosis to reconstruct dual pathways. Results The mean operation time was (240±35) minutes, the mean number of lymph nodes dissected were 22±5, and all the incised margins were negative. No anastomotic leakage, obstruction or stenosis occurred. All patients received postoperative barium meal examination. A large amount of barium directly entered the jejunum, leaving a small amount of barium entered the jejunnum via the route of remnant stomach-duodenum, and was detained in the remnant stomach for 30-60 minutes. No esophageal reflux of barium was observed. All the patients were followed up for 4-18 months, no reflux esophagitis was detected and the short-term life quality was satisfactory. Conclusions Remnant stomach-jejunal dual pathways reconstruction prevents the reflux esophagitis and dumping syndrome, preserves the pathway of duodenum and promotes the life quality of patients.
9.Effects of extract of Ginkgo biloba on apoptosis of neurocytes and expression of inducible nitricoxide synthuse after spinal cord injury in rats
Qiang AO ; Guoqiang CHEN ; Shijie WANG ; Bo XIU ; Zhaohui SUN ; Huancong ZUO
Chinese Journal of Trauma 2009;25(1):77-81
Objective To investigate the effect of extract of Ginkgo biloba (Egb) on apoptosis of nerve cells and its mechanism after spinal cord injury (SCI) in rats. Methods Forty eight adult SD rats weighing 200-230 g were divided equally and randomly into Egb group and normal saline (NS) group. After hemisectian of spinal cord at T9 vertebrae level, rats in Egb group were lavaged with 2 ml EGB (20 mg) daily and those in NS group with 2 ml NS daily. Tissue sections were collected and stained with Nissl's staining, myelin sheath staining, and inducible nitric oxide synthase (iNOS) immunohisto-chemistry as well as terminal deoxynueleotidyl transferase-mediated dUTP nick end lebeling (TUNEL) at days 1,7, 14 and 21 respectively to evaluate the injured spinal cord tissues after six rats from each group were sacrificed Results Nissl's staining manifested less swelling of the nerve cells near the injury epi-center ( rostral and caudal ), smaller cavity and demyelinated area and higher ratio of bilateral anterior horn neurons of transection side to normal side in Egb group, compared with NS group ( P <0.05). Ap-optotie index (AI) and expression of iNOS in NS group were higher than those in Egb group ( P <0.01 or P <0. 05). Furthermore, the rate of iNOS-positive cells was positively correlated with the AI (r = 0.729, P<0.01) after SCI. Conclusion Egb can prevent nerve cells from apoptosis after SCI in rats, as may be related with inhibition of expression of iNOS.
10.Primary research on the mechanisms of Xiaoyao powder influences hypothalamus-pituitary-adrenal axis of chronic stress rats
Haiqing AO ; Zhiwei XU ; Wenjun FU ; Junfang SU ; Qi SUN ; Jin HUANG ; Wenzhu WANG
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(8):680-682
Objective To observe the mechanisms that Xiaoyao powder influences the hypothalamus-pituitary-adrenal (HPA) axis of chronic-stress rats. Methods Chronic stress rats were as researching object,and RU-38486 acted as tool drugs. The serum-GC density of rats were tested with ELISA,and the glucocortcoid(GR) in hippocampus neuron were tested with immunofluorescence,the CRH mRNA in hypothalamus were tested by in situ hybridization (ISH). Results Compared with normal group ( 1.09 ± 0.11 ;0.57 ± 0.10), the expression of GR in hippocampus of model group decreased(0.65 ± 0. 10; P < 0. 01 ), and the expression of CRH mRNA in hypothalamus of model group increased ( 1.12 ±0. 11; P<0. 0l ) ,the GR in hippocampus of RU-38486 group increased ( 1.59 ± 0. 11; P < 0. 01 ), and the expression of CRH mRNA in hypothalamus of RU-38486 group reduced (0.48±0.10; P<0.05) ,but both the expression of GR in hippocampus and the CRH mRNA in hypothalamus of Xiaoyao powder group were no change (0.62 ±0.08;0.97 ±0.13; P>0.05). Compared with model group,both the expression of GR in hippocampus of RU-38486 and Xiaoyao powder group increased (P<0. 01) ,and both the expression of CRH mRNA in hypothalamus of RU-38486 and Xiaoyao powder group reduced (P<0.01). Conclusion Multi-stress can result in the expression of GR in hippocampus of rats decreasing and the expression of CRH mRNA in hypothalamus increasing, but those changes can be restrained by Xiaoyao powder, and it is the maybe mechanism of Xiaoyao Powder resisting chronic stress in HPA axis.