1.Cloning of rat neuroglobulin gene and construction of its eukaryotic expression vector
Journal of Third Military Medical University 1984;0(02):-
Objective To clone rat neuroglobulin (NGB) gene and construct its eukaryotic expression vector. Methods The total RNA was extracted from Wistar rat brain and the full length cDNA encoding NGB was obtained by RT-PCR. After the sequence was confirmed by sequencing and BLAST, it was inserted in the eukaryotic expression vector pCDNA3.1 (+), then the sequence and reading frame were confirmed by two restriction endonucleases and sequencing. Results The NGB gene was cloned with four bases mutated and its eukaryotic expression vector was constructed. Conclusion NGB expressed in Wistar rat brain. NGB gene was successfully cloned and inserted in eukaryotic expression vector.
2.Protective role of recombinant plasmid pCDNA3.1(+)/Ngb during focal cerebral ischemia of rat brain
Journal of Third Military Medical University 2003;0(11):-
Objective To observe the protective effect of recombinant plasmid pCDNA3.1(+)/Ngb during focal cerebral ischemia in rat brain.Methods Fifty-four male Wistar rats were randomly divided into three groups:normal saline(NS)control group,plasmid control group,and recombinant neuroglobulin group.NS,plasmid pCDNA3.1(+)and recombinant plasmid pCDNA3.1(+)/Ngb were respectively injected into two sites of the rat cerebra1 cortex 24 hours before induction of neocortical focal ischemia by occlusion of the right middle cerebral artery for 24 hours.The condition of local ischemic damage,expression of bcl-2 and the apoptosis in neural cells were confirmed by staining with 2% 2.3.5-triphenyltetrazolium chloride,in-site cell apoptosis detection,indirect immunofluorescent staining and Western blotting,respectively.Results The extent of cerebral infarction tissue and the apoptosis cells in the pCDNA3.1(+)/Ngb group were significantly reduced than those in other control groups(P
3.Effects of early application of pediatric amino acids on extrauterine growth and development of preterm and small for gestational age infants
Jianfang GAI ; Yong JI ; Wenyuan NIU ; Rongwei MA ; Yingru ZHAI ; Jianhong YAO
Journal of Chinese Physician 2013;15(8):1037-1040
Objective To explore the effect of early application of pediauic amino acids on extrauterine growth and development of preterm and small for gestational age infants.Methods A total of 106preterm and small for gestational age infants was selected in our NICU from June 2011 to May 2013,and randomly divided into two groups:treatment group (group T) and control group (group C).Next,each group was divided into two subgroups according to gestational age and birth weight.Such as ≥34W group (group T1,group C1),< 34W group (group T2,group C2),< 1.5 kg group (group T3,group C3) and ≥ 1.5 kg group (group T4,group C4).They were observed the effect of extrauterine growth and correlative serum biochemical indicators with application of pediatric amino acid in postnatal 6 hours and 12 hours.Resuits After 2-week treatment,the head circumference and weight growth of group T were higher than that of group C (P < 0.05,or P < 0.01),and the time of birth weight recovery was significantly shortened(P < 0.05,or P < 0.01).The incidence rate of extrauterine growth retardation(EUGR) in the group T was lower than that in group C,there is significantly statistical differences(P < 0.01).The concentration of blood urea nitrogen (BUN) in group T was obviously increased than that in the group C(P <0.01).The levels of ALT,aspartate aminotransferase (AST),total bilirubin (TBIL),cholesterol (CH) and triglycerides (TG) were not different between the group T and group C.The comparison of alanine aminotransferase (ALT) levels were statistically significant between group C4 and group T4.In the different gestational age groups,the concentration of BUN in group T was obviously higher than that in the group C after 2-week treatment(P <0.05),the levels of AST,TBIL,CH and TG were not different between the group T and group C (P > 0.05).The incidence rate of EUGR in the 4th group C was increased significantly than that in the 4th group T(P <0.05).Conclusions The preterm and small for gestational age infants were safe after the pediatric amino acid was used at the 6 h after birth.Amino acid can promote growth of head circumference and weight,shorten the birth weight recovery time and reduce the incidence of EUGR.
4.Management of obstructive hydrocephalus before posterior fossa tumor resection in children.
Wenyuan JI ; Ping LIANG ; Yudong ZHOU ; Lusheng LI ; Xuan ZHAI ; Zuozhong XIA
Journal of Southern Medical University 2013;33(11):1696-1698
OBJECTIVETo explore the management of obstructive hydrocephalus caused by posterior fossa tumors before tumor resection in children.
METHODSThe clinical data were reviewed of 162 pediatric patients of posterior fossa tumors with obstructive hydrocephalus undergoing surgical tumor removal between January 2008 and June 2012. Ninety children received preoperative Ommaya external drainage (group A) and 72 underwent preoperative ventriculo-peritoneal shunting (V-Ps) (group B). The therapeutic effects were evaluated and compared between the two groups.
RESULTSPostoperative complications found in a total of 67 cases including infection (27), shunt blockage (19), subdural hematoma or effusion (16), ventricle fissure syndrome (5), and tumor hernia (4). Significant differences were found in the incidences of shunt blockage (P=0.047) and subdural hematoma or effusion (P=0.039) but not in the incidences of intracranial infection (P=0.478) or tumor hernia (P=0.462) between the two groups.
CONCLUSIONOmmaya reservoir can produce good results through simple surgical procedures for treatment of acute hydrocephalus in children with posterior fossa tumors and is associated less trauma and complications.
Adolescent ; Astrocytoma ; complications ; surgery ; Brain Diseases ; etiology ; Child ; Child, Preschool ; Drainage ; adverse effects ; methods ; Female ; Hematoma, Subdural ; etiology ; Humans ; Hydrocephalus ; etiology ; surgery ; Infant ; Infection ; etiology ; Infratentorial Neoplasms ; complications ; surgery ; Male ; Medulloblastoma ; complications ; surgery ; Preoperative Period ; Retrospective Studies ; Ventriculoperitoneal Shunt ; adverse effects
5.Determination of non-steroidal anti-inflammatory drugs in the environmental water samples by a polyvinylimide-modified magnetic nanoparticles-based solid phase extraction coupled with high-performance liquid chromatography
Xiaohang TANG ; Huilin SONG ; Liying YAO ; Guowen QIN ; Xingchen WANG ; Wenyuan LIU ; Shunli JI
Journal of China Pharmaceutical University 2024;55(4):485-492
The long-term presence of non-steroidal anti-inflammatory drugs (NSAIDs) in the environmental water samples not only affects the life safety of aquatic organisms and disturbs the ecoenvironment, but also poses a serious threat to human health. In this study, amino-functionalized Fe3O4 nanoparticles (Fe3O4-NH2) were firstly prepared by solvothermal method. Subsequently, polyethyleneimine (PEI) with a branched chain structure was successfully grafted onto Fe3O4 nanoparticles by Schiff base reaction in aqueous solution at room temperature using glutaraldehyde as a cross-linking agent, and a recyclable PEI-grafted magnetic nano-sorbent (Fe3O4@PEI) was synthesized and applied for the detection of NSAIDs in the environmental water samples. The compositional properties of Fe3O4@PEI were investigated by different characterization methods and the parameters affecting the extraction of NSAIDs were optimized. Due to high adsorption of Fe3O4@PEI for NSAIDs, the quantitative analysis of four NSAIDs in the environmental water samples, ketoprofen, naproxen, diclofenac and tolfenamic acid, was performed in combination with high-performance liquid chromatography. A good linear relationship between the chromatographic peak area and concentration was observed in the range of 1−500 µg/mL. The recoveries of the samples at three different spiked levels ranged from 85.6% to 107.8%; the intra-day precision was less than 7.8% (n=6); and the inter-day precision was less than 9.5% (n=3). The method is simple, rapid, accurate and reliable, and can be used for the analysis of NSAIDs in the environmental water samples.
6.Inhibition of islet allograft rejection by Qa-1/PD-L1 artificial liposome
Meng GUO ; Yuanyu ZHAO ; Hao YIN ; Jiayong DONG ; Junsong JI ; Qi LU ; Hang YUAN ; Fei TENG ; Wenyuan GUO ; Guoshan DING
Chinese Journal of Organ Transplantation 2019;40(2):72-77
Objective To explore the effects of Qa-1 and PD-L1 loaded artificial liposomal treatment in allograft rejection and its outcomes .Methods The extracellular domains of Qa-1 and PD-L1 were loaded on liposome surface by streptavidin-biotin system . Mixed lymphocyte reaction (MLR) was performed for measuring Qa-1/PD-L1 liposome biological function .Then liposome was co-transplanted with allo-islets via portal vein .The levels of blood glucose and C-peptide were detected daily after transplantation .Also hepatic lymphocytes after transplantation were isolated for determining the proportion of activated cells and signaling pathway changes .Results Artificial liposome could be easily loaded with biotinylated peptide and its diameter was between 50 to 500 nm . Qa-1/PD-L1 liposome could significantly suppress lymphocyte proliferation , activation and secretion of IFN-γ in MLR by an activation of SHP1/2 and an inhibition of Syk pathway .Qa-1/ PD-L1 liposomes could suppress the activation of hepatic lymphocytes in vivo by activating SHP1/2 ,protecting islet allografts and maintaining a normal level of blood glucose in recipients .Conclusions Qa-1/PD-L1 loaded liposome can effectively suppress allograft rejection and improve the outcomes of islet transplantation .
7.Effects of Urtica dioica extract on malignant biological behaviors of breast cancer cells and its possible mechanism
JI Wenyuan ; WEI Shaoyin ; LIU Wei
Chinese Journal of Cancer Biotherapy 2021;28(8):803-809
[摘 要] 目的:探讨大荨麻提取物对乳腺癌细胞增殖、凋亡和细胞周期的影响,并初步探讨其可能的作用机制。方法:用不同质量浓度的大荨麻提取物(0、1、2、4、8、16、32、64 mg/ml)处理乳腺癌细胞MCF-7和MDA-MB-231 24 h,MTT法检测细胞增殖活力,选择中位抑制浓度附近的浓度(5和10 mg/ml)作为给药浓度分别处理MCF-7和MDA-MB-231细胞24 h后,平板克隆形成实验和流式细胞术分别检测大荨麻提取物对乳腺癌细胞增殖、周期和凋亡的影响,WB法检测对细胞周期和凋亡相关蛋白以及PI3K/AKT信号通路相关蛋白表达的影响。在MCF-7细胞用5 mg/ml大荨麻提取物处理的同时转染过表达AKT质粒(大荨麻+AKT组),转染空载质粒为对照组(大荨麻+vec组),WB法检测过表达效率,比较过表达AKT对细胞增殖、周期和凋亡的影响。结果:各大荨麻提取物处理组MCF-7和MDA-MB-231细胞增殖活力均显著低于对照组(P<0.05或P<0.01);与对照组比较,5或10 mg/ml大荨麻处理组乳腺癌细胞的克隆形成数显著减少,G0/G1期细胞占比和凋亡率显著增加(P<0.05或P<0.01),P21、BAX蛋白表达显著升高而Cyclin D1、CDK4、Bcl2蛋白以及p-PI3K、p-AKT蛋白表达显著降低(P<0.05或P<0.01)。大荨麻+AKT组p-AKT和AKT蛋白表达显著高于大荨麻+vec组,克隆数、S期和G2/M期细胞占比均高于大荨麻+vec组(P<0.05或P<0.01),G0/G1期细胞占比和凋亡率低于大荨麻+vec组(P<0.05或P<0.01)。结论:大荨麻提取物可以抑制乳腺癌细胞增殖、促进凋亡且阻滞细胞在G0/G1期,其作用机制可能与抑制PI3K/AKT信号通路相关。