1.Effect of peroxisome proliferator-activated receptor-gamma excitomotor on the expression of nuclear factor-kappa B and apoptosis of retinal ganglion cells in rat retina with diabetes mellitus
Wen-jun, GOU ; Ke, OU-YANG ; Hong-bin, LV ; Qing-lan, LI ; Qi, ZHOU ; Jun, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;30(8):709-714
Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P<0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P> 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P<0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P<0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P<0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P<0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.
2.PRELIMINARY STUDY ON AN ANTIBIOTIC-PRODUCING BACTERIUM
Xi-Qian LAN ; Jun-Hua HU ; Hong-Xiu WEN ; Jia-Lian CHEN ; Ze-Yang ZHOU ;
Microbiology 1992;0(05):-
An antibiotic-producing bacterium, which was numbered as 20 #-5, was separated from the soil in Chongqing. It was identified as the member of pseudomonas. Gram positive bacteria are badly suppressed by it. The antibiotic secreted by 20 #-5 can endure 100℃ for half an hour, and it can also go through the ultrafiltration membrane with pores of 0.22?m.
3.Regulation of microRNA-199a on adhesion, migration and invasion ability of human endometrial stromal cells
Lan DAI ; Liying GU ; Jie ZHU ; Jun SHI ; Yao WANG ; Fang JI ; Wen DI
Chinese Journal of Obstetrics and Gynecology 2011;46(11):817-821
Objective To study the regulation of microRNA 199a (miR-199a) on adhesion,migration and invasion ability of human eutopic endometrial stromal cells (ESC) from patients with endometriosis.Methods ESC were transfected with miR-199a mimics or negative control (NC) RNA by lipofectamine 2000.The adhesion,migration and invasion ability of ESC were detected by cell adhesion assay,scratch assay,cell migration assay and matrigel invasion assay,respectively.Luciferase reporter assay was used to evaluate whether IKKβ was the target gene of miR-199a.The expression of ikappa B kinase beta (IKKβ),inhibitory kappa B alpha (IκB-α),phospho-IκB-α (p-IκB-α) and nuclear factor-kappa B (NF κB) protein were measured by western blot.Results ( 1 ) Adhesion potential:the adhesion inhibitory rates were ( 14 ± 4 )% in miR-199a group and 0 in control group,which showed significant difference (P<0.01 ).(2) Migration and invasion:in the scratch assay,ESC transfected with miR-199a exhibited a lower scratch closure rate than that of controls.In migration and invasion assays,the migration and invasion ability of miR-199a group were significantly decreased compared with those of NC group [ 130 ± 31 vs.247±36 (P<0.01); 63 ± 15 vs.133 ± 17 (P<0.01),respectively].(3) The luciferase activity of miR-199a group was significantly lowered than that of control group [ 0.160 ± 0.006 vs.0.383 ± 0.083 ( P <0.01 ) ].The protein levels of IKKβ,p-IκB-α,IκB-α and NF-κB of 0.350 ±0.195,0.443 ±0.076,1.970 ±0.486 and 0.454 ± 0.147 in miR-199a group were significantly different compared with the NC group in which the protein levels were set at 1.000 ( P < 0.01 ).Conclusions miR-199a can inhibit the adhesion,migration and invasion of the ESC.IKKβ is the target gene of miR-199a in ESC.One of the mechanisms of the inhibition effect is probably that miR-199a inhibits the activation of NF-κB signaling pathway by targeting IKKβ gene.
4.Cytotoxic effects of cytokine-induced killer cells transfected with the interleukin-2 gene on malignant melanoma cells
Lan LU ; Conghua XIE ; Haozhong ZHANG ; Lyuye XU ; Xingwei SHI ; Jun XIE ; Biao CHE ; Wen DING
Chinese Journal of Dermatology 2017;50(4):257-262
Objective To evaluate cytotoxic effects of cytokine-induced killer cells (CIK cells) transfected with the interleukin-2 (IL-2) gene on malignant melanoma cells.Methods Mouse spleen cells were extracted,lymphocyte cells were separated,and CIK cells were prepared from these lymphocyte cells.PEGF-N1 plasmids containing IL-2 gene (PEGF-NI-IL-2) were transfected into CIK cells.Fluorescence microscopy was used to observe transfection products,and reverse transcriptase-polymerase chain reaction (RT-PCR) was conducted to determine the IL-2 mRNA expression.Then,effector cells such as CIK cells and IL-2-transfected CIK cells were separately co-cultured with target cells (B16 melanoma cells) at effector-target ratios of 10∶ 1,20∶1 and 40∶1,then 4-hour lactate dehydrogenase release assay was performed to evaluate cytotoxic effects of the two kinds of CIK cells on B 16 cells.After effector cells were cocultured with target cells at the effector-target ratio of 40∶1 for 48 hours,enzyme-linked immunosorbent assay (ELISA) was conducted to detect levels of IL-2,interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the supernatant of the two kinds of CIK cells.Finally,mouse models of melanoma were established,and a total of 28 melanoma-bearing mice were divided into 4 groups to be peritumorally injected with 0.2 ml sodium chloride physiological solution (control group),100 IU IL-2 solution (IL-2 group),CIK cell suspension at a cell density of 1 × 106 cells per milliliter (CIK group) and IL-2-transfected CIK cell suspension at a cell density of 1 × 106 cells per milliliter (IL-2-transfected CIK group) respectively.Tumor morphology,tumor inhibition rate and cell apoptosis rate were used to evaluate tumor growth in the above groups.If data were normally distributed,t-test was used for comparing means between two groups,and analysis of variance and least significant difference (LSD)-t test were used for comparing means among multiple groups.Results Fluorescence microscopy and RT-PCR both showed that IL-2 was successfully transfected into CIK cells.The cytotoxic effect of IL-2-transfected CIK cells on B16 cells was strongest at the effector-target ratio of 40:1.Levels of IL-2,IFN-γ and TNF-α were also significantly higher in the supernatant of IL-2-transfected CIK cells [(1107.26 ± 6.49) pg/ml,(50.01 ± 3.35) pg/ml,(39.86 ± 3.25) pg/ml] than those in that of CIK cells [(51.09 ± 3.85) pg/ml,(32.71 ± 2.43) pg/ml,(30.11 ± 3.08) pg/ml,t =442.60,14.93,6.89,all P < 0.01].Animal experiments showed that the tumor volume obviously increased in the control group (P < 0.05),but significantly decreased in the IL-2 group,CIK group and IL-2-transfected CIK group (all P < 0.001) after intervention compared with those before intervention.Furthermore,the tumor volume in the IL-2-transfected CIK group was significantly less than that in the other three groups (all P < 0.01),but no significant difference was observed between the IL-2 group and CIK group (P > 0.05).In addition,the apoptosis rate was significantly higher in the IL-2 group,CIK group,and IL-2-transfeeted CIK group than that in the control group (all P < 0.01).The apoptosis rate and tumor inhibition rate were significantly higher in the IL-2-transfected CIK group than those in the IL-2 group and CIK group (all P < 0.01),but insignificantly different between the IL-2 group and CIK group (P > 0.05).Conclusion IL-2-transfected CIK cells had stronger killing effects on malignant melanoma.
5.Bone Marrow Mesenchymal Stem Cells Differentiated into Hepatocyte-like Cells Induced by Rebirth Liver Tissue Lixivium of Mouse in Vitro
Jun-lan YU ; Rong LI ; Wen-jie WANG ; Li LI ; Juchao LIU ; Jinjing WANG ; Yingxin XU
Chinese Journal of Rehabilitation Theory and Practice 2006;12(12):1056-1057
Objective To induce bone marrow mesenchymal stem cells (BMSCs) differentiated into hepatocyte-like cells with rebirth liver tissue lixivium of mouse in vitro.Methods Mouse BMSCs were isolated and directionally induced with rebirth liver tissue lixivium of mouse 36 h after partial hepatectomy (PH). The morphology of cells was observed under an invert microscope, and the characteristics of differentiated cells was identified by immunofluorescence test.Results 7 days after induced by liver lixivium, the spindle shaped BMSCs became round and resembled hepatocyte-like cells. 1~2 weeks later, the differentiated cells expressed albumin, CK8 and CK18, which were known as characteristic markers of the hepatocyte.Conclusion BMSCs of mouse can be differentiated into hepatocyte-like cells under induction of liver tissue lixivium of mouse.
6.Effect of Qidan Granule on PMC Derived Peptide Content and Structure of Hippocampal CA1 Region in Microwave Radiated Rats.
Lan-fang GUAN ; Ya-wei LI ; Jun-jie XU ; Xiu-hong ZHONG ; Hong ZHANG ; Wen-he ZHU ; Shi-jie LV
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(4):471-475
OBJECTIVETo explore the protection of high intensity microwave radiation on hypothalamo-pituitary-adrenal axis (HPAA) activity and hippocampal CA1 structure in rats and the protectiveeffect of Qindan Granule (QG) on radiation injured rats.
METHODSTotally 48 Wistar rats were randomlydivided into 8 groups, i.e., the normal control group, post-radiation day 1, 7, and 10 groups, 7 and 10days prevention groups, day 7 and 10 treatment groups, 6 in each group. Rats in prevention groups wererespectively administered with QG liquid (1 mL/100 g, 4. 75 g crude drugs) for 7 days and 10 days bygastrogavage and then microwave radiation. Then preventive effect for radiation injury was statisticallycalculated with the normal control group and the post-radiation day 1 group. Rats in treatment groupswere firstly irradiated, and then administered with QG liquid (1 mL/100 g, 4.75 g crude drugs). Finally preventive effect for radiation injury was statistically calculated with the normal control group, post-radiation day 7 and 10 groups. Contents of corticotrophin releasing hormone (CRH), beta endorphin (beta-EP), adrenocorticotropic hormone (ACTH), and heat shock protein 70 (HSP70) were detected. Morphological changes and structure of hippocampal CA1 region were observed under light microscope.
RESULTSCompared with the normal control group, contents of CRH and beta-EP significantly decreased in each radiation group. Serum contents of ACTH and beta-EP significantly increased in post-radiation day 1 and 7 groups (P < 0.05). Compared with radiation groups, beta-EP content in serum and pituitary significantly increased, and serum ACTH content significantly decreased in prevention groups (P < 0.05). Pituitary contents of CRH and beta-EP significantly increased in prevention groups. Serum contents of ACTH, beta-EP, and HSP70 were significantly lower in day 7 treatment group than post-radiation day 7 group (P < 0.05). Morphological results showed that pyramidal neurons in the hippocampal CA1 region arranged in disorder, with swollen cells, shrunken and condensed nucleus, dark dyeing cytoplasm, unclear structure. Vessels in partial regions were dilated with static blood; tissues were swollen and sparse. In prevention and treatment groups pathological damage of hippocampal CA1 region was obviously attenuated; neurons were arranged more regularly; swollen, pycnotic, or deleted neuron number were decreased; vascular dilatation and congestion was lessened.
CONCLUSIONQG could affect HPAA function and activity of high intensity microwave radiated rats, showing certain preventive and therapeutic effects of microwave radiated rats by adjusting synthesis and release of partial bioactive peptides and hormones in HPAA, improving pathological injury in hippocampal CA1 region.
Adrenocorticotropic Hormone ; blood ; Animals ; CA1 Region, Hippocampal ; drug effects ; pathology ; radiation effects ; Corticotropin-Releasing Hormone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; HSP70 Heat-Shock Proteins ; blood ; Hypothalamo-Hypophyseal System ; drug effects ; radiation effects ; Microwaves ; adverse effects ; Pituitary-Adrenal System ; drug effects ; radiation effects ; Random Allocation ; Rats ; Rats, Wistar ; beta-Endorphin ; blood ; metabolism
7.A new chromone glycoside from roots of Polygonum multiflorum.
Hui-Nan ZHAO ; Li-Li CHEN ; Xiao-Jun HUANG ; Ying WANG ; Yao-Lan LI ; Wen-Cai YE
China Journal of Chinese Materia Medica 2014;39(8):1441-1444
Several kinds of column chromatography methods were used to investigate the chemical constituents of roots of Polygonum multiflorum. The structures of the isolated compounds were identified based on their physicochemical properties, spectral data and chemical methods. A new chromone glycoside was isolated and its structure was identified as (S)-2-(2'-hydroxypropyl)-5-methyl7-hydroxychromone-7-0-alpha-L-fucopyranosyl (1-->2)-beta-D-glucopyranoside (1).
Drugs, Chinese Herbal
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chemistry
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Glycosides
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chemistry
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Molecular Structure
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Plant Roots
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chemistry
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Polygonum
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chemistry
8.Alterations in cardiac structure and function in a modified rat model of myocardial hypertrophy.
Wen-Jun, DAI ; Qi, DONG ; Min-Sheng, CHEN ; Lu-Ning, ZHAO ; Ai-Lan, CHEN ; Zhen-Ci, LI ; Shi-Ming, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(5):626-33
This study was aimed to establish a stable animal model of left ventricular hypertrophy (LVH) to provide theoretical and experimental basis for understanding the development of LVH. The abdominal aorta of male Wistar rats (80-100 g) was constricted to a diameter of 0.55 mm between the branches of the celiac and anterior mesenteric arteries. Echocardiography using a linear phased array probe was performed as well as pathological examination and plasma B-type natriuretic peptide (BNP) measurement at 3, 4 and 6 weeks after abdominal aortic constriction (AAC). The results showed that the acute mortality rate (within 24 h) of this modified rat model was 8%. Animals who underwent AAC demonstrated significantly increased interventricular septal (IVS), LV posterior wall (LVPWd), LV mass index (LVMI), cross-sectional area (CSA) of myocytes, and perivascular fibrosis; the ejection fraction (EF), fractional shortening (FS), and cardiac output (CO) were consistently lower at each time point after AAC. Notably, differences in these parameters between AAC group and sham group were significant by 3 weeks and reached peaks at 4th week. Following AAC, the plasma BNP was gradually elevated compared with the sham group at 3rd and 6th week. It was concluded that this modified AAC model can develop LVH, both stably and safely, by week four post-surgery; echocardiography is able to assess changes in chamber dimensions and systolic properties accurately in rats with LVH.
9.A Corpus-Based Stylistic Analysis on Structured Abstract of Literature Review Papers
lan-ying, WANG ; lian-zhu, WANG ; jia-qin, WANG ; rui-jun, ZHANG ; wen-ming, YONG ; hui-shao, ZHENG
Journal of Applied Clinical Pediatrics 2006;0(23):-
To introduce the writing techniques for structured Abstract of literature review papers based on the knowledge of corpus linguistics,so as to summarize the stylistic,structural,syntactic and lexical characteristics,objectively and systematically.This research will also faci-litate the authors and translators,who have difficulties in writing structured English Abstract,to publish their articles via mastering its stylistic characteristics,writing format and language.
10.Efficacy and safety of pre-hospital use of hemagglutinin in patients with craniocerebral trauma and scalp laceration
Hua-Lei TANG ; Lan YI ; Pei-Wen ZHANG ; Shan PENG ; Wen-Jun CHEN
Journal of Regional Anatomy and Operative Surgery 2018;27(4):300-303
Objective To discuss the efficacy and safety of pre-hospital use of hemagglutinin in patients with craniocerebral trauma and scalp laceration.Methods Totally 100 patients with craniocerebral trauma and scalp laceration who were admitted into our hospital from De -cember 2014 to December 2017 were selected as the object of study.These patients were divided into the hemagglutinin group and the control group with 50 cases in each group according to the random number table method.The control group was given conventional first aid treatment, while the hemagglutinin group was given pre-hospital use of hemagglutinin on the basis of conventional first aid treatment.Then,the efficacy and safety of the two groups were compared.Results The hemoglobin(Hb),hematocrit(HCT),platelet aggregation rate(PAR)of the he-magglutinin group were significantly higher than those of the control group after treatment,and the difference was statistically significant(P<0.05).The prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(FIB)levels and embolism complication rates of the hemagglutinin group and the control group were basically the same after treatment,and the difference was not statistically significant(P>0.05).The hypotension rate and poor prognosis of Glasgow outcome score(GOS)of the hemagglutinin group were significantly lower than those of the control group with statistically significant difference(P<0.05).Conclusion Pre-hospital use of hemagglutinin can effectively improve the hemostatic effect in patients with traumatic brain injury and scalp laceration,and it is beneficial to improve the hypotension and prognosis of patients with high safety.