1.Vinpocetine alleviates cerebral ischemia-reperfusion injury in rats by regulation of the expressions of nuclear factor κB p65, peroxisome proliferator-activated receptor γ and cyclooxygenase-2
Xiaobo QIU ; Jian WANG ; Lanying HE ; Yong LUO
International Journal of Cerebrovascular Diseases 2015;23(7):517-521
Objective To investigate neuroprotective mechanisms of vinpocetine by observing the effects of vinpocetine injection on the expressions of peroxisome proliferators-activated receptor γ(PPARγ),nuclear factor (NF)-κB p65,cyclooxygenase-2 (COX-2) in the ischemic cortex,and infarct volume after focal cerebral ischemia-reperfusion in rats.Methods A focal cerebral ischemia-reperfusion injury model was induced by suture method.The rats were randomly divided into a normal control,a cerebral ischemiareperfusion and a vinpocetine groups.They were also divided into either a day 7 subgroup or a day 14 subgroup (n =6 in each subgroup) according to the reperfusion time.Western blot was used to detect the expression levels of PPARγand NF-κB P65 in the ischemic cortex.Triphenyl tetrazolium staining was used to detect the volume of cerebral infarction.Results Western blot showed that at day 7 and 14 after cerebral ischemia-reperfusion,expression levels of PPARκ (all P<0.001) and NF-κB p65 (all P<0.001) in the cerebral ischemia-reperfusion group were significantly higher than those in the sham operation group,the expression levels of PPARκ (all P <0.05) in the vinpocetine group were significantly higher than those in the cerebral ischemia-reperfusion group,but the expression levels of NF-κB p65 (all P <0.05) were significantly lower than those in the cerebral ischemia-reperfusion group.Reverse transcription polymerase chain reaction showed that COX-2 mRNA expression levels were upregulated significantly at day 7 and 14 after cerebral ischemia-reperfusion compared with the sham operation group (all P < 0.001),the expression levels of COX-2 mRNA in the vinpocetine group were significantly downregulated compared with the cerebral ischemia-reperfusion group (all P< 0.05).The infarct volumes at day 7 (134.308± 9.954 mm3vs.185.543 ± 9.100 mm3;q=10.659,P<0.001) and at day 14 (137.865 ± 9.094 mm3vs.183.210±4.368 mm3;q=11.166,P<0.001) in the vinpocetine group were significantly less than those in the cerebral ischemia-reperfusion group.Conclusions Vimpocetine significantly reduces infarct vohme after focal cerebral ischemia-reperfusion,its mechanism may be associated with upreguhtion of PPARγexpression and downreguhtion of the expressions of NF-κB p65 and COX-2.
2.STUDY ON THE EFFECT OF IODINE EXCESS ON CATHEPSINS IN RAT THYROID CELLS
Xiaoli LIU ; Jian XU ; Lanying LI ; Yun SUN ; Liping HAO ; Xiufa SUN
Acta Nutrimenta Sinica 2004;0(05):-
Objective: To explore the effect of iodine overdose on cathepsin B and D in thyroidcell line (FRTL). Method: The cells were treated with potassium iodide (0, 1, 5, 10, 50 and 100mmol/L). After treatment 6, 12, 24, 48 and 72 hours, the cells were collected. The activities of cathepsin B (CB) and cathepsin D (CD) were assayed by fluorospectrophotometry and Lowry, respectively. The mRNA levels of CB and CD were determined with real-time PCR. Results: After 12 and 24 hours, the activities of CB and CD, treated by 50mmol/L potassium iodide were decreased significanty. The mRNA levels of CB and CD of 50mmol/L potassium iodide group were decreased significanty after treatment 24 hours. Conclusion: The decreased activities of CB and CD were one of reasons of thyroid colloid retention resulted from iodine excess.
3.THE INTERVENTION EFFECT OF SELENIUM ON DECREASE OF CATHEPSIN B,D ACTIVITIES OF THYROID FOLLICULAR EPITHELIAL CELL LINE INDUCED BY IODINE EXCESS
Jian XU ; Xiaoli LIU ; Xuefeng YANG ; Lanying LI ; Yun SUN ; Xiufa SUN
Acta Nutrimenta Sinica 2004;0(05):-
Objective: To detect the damage of excessive iodine on the activities and mRNA of cathepsin B and D and observe the effects of selenium intervention. Method: Thyroid follicular epithelial cell line FRTL was cultured in Kaighn’s modified Ham’s F-12 medium. ① The cells were divided into 8 groups for detection of activities of cathepsin B and D (CB and CD). The medium of NI group did not contain selenium or iodine, while EI group contained 50 mmol/L KI, and IS group contained 50mmol/L KI and different dosages of 0.1,0.2,0.5,1.0,2.0 and 5.0 ?mol/L selenium (sodium selenite). ②The mRNA levels of cathepsin B and D were detected by real-time PCR. The medium of NI group and EI group was same as above, but IS group only contained 0.1?mol/L selenium besides 50mmol/L KI. Results: The activities of CB decreased in EI group (50mmol/L KI treated 12 h), while CD decreased after 24 h and both activities were improved in IS groups with 0.1~0.5?mol/L selenium. The mRNA levels of CB and CD in EI group decreased, while in IS group were improved. Conclusion: Excessive iodine resulted in goiter through decreased levels of activities and mRNA of CB and CD. The intervention of selenium on goiter mayinduce its amelioration effect on decrease of CB and CD activities and the optimal dosage ranges from 0.1?mol/L to 0.5?mol/L.
4.A study of inductive factors of embryonic stem cells differentiating into cardiac myocytes in vitro
Lanying LIU ; Kun YANG ; Zhengyu ZHU ; Yuchuan LIU ; Xiyong YU ; Jian TANG ; Wei YIN ; Jianquan MA ; Jun GU
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To study cellular and molecular mechanisms of cardiac development associated genes ex- pression and its function during early stage cardiomyogenesis. METHODS: (1 ) Mouse embryonic stem cells (ESC) line D3 culture. (2) Inductive culals of ESC differentiated into cardiac myocytes in vitro.(3) Identification of ESC -derived cardiac myocytes: RNA isolation; synthesis of specific primer and RT - PCR; Label of RT - PCR products with [? - 32P] dATP as probes, purifyed by sephadex G - 50 columns, determined the yield of DNA. RNA dot hy- bridization. RESULTS: 80% of ESC differentiated into cardiomyocytes by improved conditional medium. Cardiomy- ocytes contraCted in a synchronous manner. The results of RT - PCR and RNA blot showed that cardiac genes were expressed abundantly and specifically during the early cardiomyogenesis. CONCLUSIONS: ESC were able to be dif- ferentiate into cardiomyocytes. Different concentrations and components of RA, DMSO and FCS affected ESC car- diomyogenesis in de. The optimal result obtained was from the conditional medium, a mixturce of 2 nmol/L retinoic acid (RA), 0.6% dimethyl sulfoxide (DMSO) and 20% fend calf serum (FCS).
5.Clinical features and nursing strategy of 8 patients with rhinocerebral mucormycosis
Lanying WEN ; Jian′guo HUANG ; Fengjiao YAN
Chinese Journal of Modern Nursing 2015;21(6):686-688
Objective To study the clinical features and nursing strategy of rhinocerebral mucormycosis , and to improve the knowledge of the disease and the quality of care .Methods The data of 8 cases with rhinocerebral mucormycosis were reviewed and analyzed .The clinical features and nursing intervention were analyzed throughout the course of disease .Results The underlying immunosuppressive condition of the 8 cases of mucormycosis had diabetes , diabetic ketoacidosis , anaemia or adiotherapy with headache and orbitial apex syndrome as the early symptoms .Nurses focused on local signs observation , perioperative change of intracranial pressure and general conditions , drug adverse reaction observation and nursing care of original disease , mental care, and basic nursing care .There were certain effects for corresponding nursing measures without occurrence of nursing events duration of hospital stay .1 patient died within one week in hospital;4 patients had survived from 3 to 12 months; and 3 patients had been alive over 3 years until now.Conclusions Rhinocerebral mucormycosis was the secondary onset severe diseases for patients with poor immunologic function , implicated in the orbit and brain , and the fatality rate was high .It could hold the development of the disease by clearing up the focus and wiping off the condition of fungi thrive together with medical management and adequacy nursing intervention .
6.Distribution and significance of CD1a+and CD83+dendritic cells in lung tissue of COPD mice
Lanying ZHANG ; Fuan ZHANG ; Maomao LIU ; Jie CHEN ; Jian ZHOU ; Yuting LIU ; Yao OUYANG
Tianjin Medical Journal 2024;52(9):913-916
Objective To study the distribution and significance of CD1a and CD83 positive dendritic cells(DCs)in lung tissue of chronic obstructive pulmonary disease(COPD)mice.Methods Twenty C57BL/6 mice were randomly divided into the air control group and the smoked COPD group(n=10 for each group).COPD mouse model was established using cigarette smoking method.Mice were executed within 24 h after the last cigarette smoking,and right lower lung was collected.Body mass changes and lung histopathological changes of mice were observed in two groups.Mean linear intercept(MLI)was measured,and expression levels of CD1a+and CD83+DCs in lung tissue were detected by immunohistochemistry.Results The body mass of mice at 7,14,21 and 28 d after modeling was lower in the smoked COPD group than that in the air control group(P<0.05).HE staining showed that the normal alveolar structure of lung tissue of mice in the smoked group was disrupted,with multiple alveoli fused with each other to form a larger alveolar lumen,a large number of inflammatory cells infiltrated in alveolar intervals,and walls of the alveoli were thickened.COPD modeling was successful.Compared with the air control group,MLI values(μm)increased in the smoked COPD group(28.30±3.47 vs.50.40±3.60),and the number of CD1a+DCs(per field of view)in lung tissue increased(9.58±2.18 vs.17.08±3.67),while the number of CD83+DCs(per field of view)decreased(19.78±4.95 vs.8.02±3.30)(all P<0.05).Conclusion The number of CD1a+DCs in lung tissue is increased and the number of CD83+DCs in lung tissue is decreased in the smoked COPD group of mice,and cigarette smoking may have impaired DC maturation.
7.miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1
Shixiang SU ; Yuyang WANG ; Zongshuai QIN ; Guixiang HUANG ; Jian XU ; Lanying CEN ; Yueqiu QIN
Basic & Clinical Medicine 2024;44(1):23-30
Objective To investigate the effect of miR-142-3p on the apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1.Methods AR42J cells were divided into blank group(blank),acute pancreatitis model group(AP,100 nmol/L cerulein for 24 h),and then transfected with miR-142-3p mimics,mimics NC,miR-142-3p inhibitor and inhibitor NC,respectively.The cells in the model group were recorded as miR-142-3p mimics group,mimics NC group,miR-142-3p inhibitor group and inhibitor NC.The expression of miR-142-3p in cells was detected by RT-qPCR.The protein expressions of HMGB1,caspase-3,Bax and Bcl-2 were detected by Western blot.Hoechst staining was used to determine cell apoptosis.The apoptosis rate of cells was detected by flow cytometry.The targeting relationship between miR-142-3p and Hmgb1 was determined by dual luciferase reporter gene assay.Results Compared with blank control group,the expression level of miR-142-3p in the AP group was significantly down-regulated(P<0.01),the expression level of HMGB1 and caspase-3 proteins was up-regulated(P<0.05),the expression level of Bax protein was significantly up-regulated(P<0.01),the expression level of Bcl-2 protein was significantly decreased(P<0.01)and the apoptosis rate increased significantly(P<0.01).Compared with the mimics NC group,the level of miR-142-3p in the miR-142-3p mimics group was significantly up-regulated(P<0.01),the expression of HMGB,caspase-3 and Bax proteins was significantly down-regulated(P<0.01),the expression of Bcl-2 protein was up-regulated(P<0.05),and the apoptosis rate decreased signifi-cantly(P<0.01).Compared with inhibitor NC group,the expression level of miR-142-3p in miR-142-3p inhibitor group was down-regulated(P<0.05),the expression levels of HMGB1,caspase-3 and Bax proteins were signifi-cantly up-regulated(P<0.01),the expression level of Bcl-2 protein was decreased(P<0.05)and the apoptosis rate increased significantly(P<0.01).The dual luciferase reporter gene assay showed that Hmgb1 was the target gene of miR-142-3p.Conclusions 1)The expression of miR-142-3p was low in the model group.2)miR-142-3p can inhibit the apoptosis of AR42J cells by inhibiting the expression of Hmgb1.
8.Experimental study on the in vitro induction of regulatory T cells by umbilical cord mesenchymal stem cells with positive human leukocyte antigen-G
Jian BAI ; Li XIAO ; Lanying MIAO ; Dayong LIN ; Hong LIU ; Yu GAO ; Wen CHEN ; Lili BI ; Xiangrui KONG ; Haiyan HUANG ; Bingyi SHI
Organ Transplantation 2018;9(2):97-102
Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.
9.Application value of different language assessment tools in language development assessment of 12-month-old high-risk infants
Lanying JIAN ; Yan LUO ; Yan ZHU ; Chengwei SHEN ; Kexin TU ; Huiling QIN ; Lifang JIN
Chinese Journal of Child Health Care 2024;32(2):149-153
【Objective】 To explore the application value of different language assessment tools in the assessment of language development of 12-month-old high-risk infants, and to screen out simple and valid language assessment tools. 【Methods】 A total of 217 11- to 13-month-old high-risk infants who were followed up at the outpatient service for high-risk infants at the child health clinic of Guiyang Maternal and Child Health Hospital from March 2022 to May 2023 were selected as the study subjects. Their language was evaluated by Early Language Milestone Scale (ELMS), Putonghua Communicative Development Inventory (PCDI) and Ages and Stages Questionnaire-Third Edition (ASQ-3). With Gesell as the gold standard for the assessment of language, the area under receiver operating characteristic curve (AUC), sensitivity, specificity, accuracy, positive predictive value, negative predictive value, Youden index and Kappa value of the three tools were calculated. Spearman correlation analysis was used to analyze the correlation between the different language assessment scales. The Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) was used to evaluate the three tools. 【Results】 1)Among 217 high-risk infants, 78 preterm infants was the most (35.94%). The rate of delayed language development detected by Gesell, ELMS, PCDI-comprehension, PCDI-expression, ASQ-3 were 5.5%, 7.8%, 36.4%, 30.0% and 11.5%, respectively. 2)ASQ-3 had the strongest correlation with Gesell language region (r