2.Electrophysiological characteristics of early-differentiated neural stem cells in embryonic rats
Jie LENG ; Li JIANG ; Hengsheng CHENG
Journal of Third Military Medical University 1983;0(03):-
Objective To study the electrophysiological characteristics of early-differentiated neural stem cells(NSC)in embryonic rats.Methods NSC were isolated from hippocampus of embryonic rats and cultured.The cells were plated onto poly-llysine hydrochloride glass cover-slips for 1 h or differentiated for 1-3 d in a differentiation medium.Whole cell patch-clamp technique was used to record the voltage-gated ion channel current.Results No inward Na+ current was detected in any undifferentiated NSC.One day after induction of differentiation,the cells exhibited a voltage-gated Na+ current,but could not induce an action potential.Two types of outward K+ current(delayed rectifier K+ and transient K+ current) and inward T-type Ca2+ current were detected in NSC and early-differentiated NSC.Conclusion The functional expression of Na+ current is a marker of early differentiation of NSC following cell cycle exit.Early differentiation of NSC is an important development stage of their ion channels.
3.Effect of morphine on synaptic long-term potentiation in spinal dorsal horn evoked by electric stimulation of sciatic nerve in rats
Jiang WU ; Deying HUANG ; Jie CHENG ; Shouqin SHANGGUAN ; Qisheng HU
Chinese Journal of Anesthesiology 2009;29(4):346-348
Objective To evaluate the effect of morphine on synaptic long-term potentiation (LTP) in the spinal dorsal horn evoked by electric stimulation of sciatic nerve in rats. Methods Twenty-seven healthy male SD rats aged 60-90 d weighing 180-200 g were randomly divided into 4 groups: group Ⅰ control (group C, n=7), group Ⅱ morphine (group M, n=7), group Ⅲ naloxone (group N, n=6), and group Ⅳ morphine + naloxone (group MN, n=7). The animals were anesthetized with intraperitoneal 10% urethane 1 g/kg, intubated and then mechanically ventilated. The bipolar insulated stainless steel recording electrode (impedance 0.5-1 MΩ, diameter 0.1 mm) was inserted into the left side of the spinal dorsal horn at T13-L1 to stimulate the left side of the sciatic nerve. Single square pulses (15 V, 0.5 ms, 1/60 Hz for 30 min) was applied to evoke spinal field potentials. Normal saline 10 μl, morphine 10 μl (15 μg/μl), naloxone 10 μl (2.5 μg/μl), and the mixture 10 μl of naloxone 5 μl (2.5 μg/μl) and morphine 5 μl (15 μg/μl) was gradually instilled over 2 rain in the 4 groups respectively. Five minutes later, high-frequency and intensity tetanic stimulation (30-40 V, 0.5 ms, 100 Hz, given in 4 trains of 1-s duration at 10-s intervals) was used to induce LTP. Then single square stimuli (15 V, 5 ms, 1/60 Hz) were applied to the sciatic nerve for 210 min. The amplitude and latency period of the field potential were recorded 30 min before tetanic stimulation, and 0-30, 35-60, 65-120 and 125-210 min after titanic stimulation. Results Compared with group C, the amplitude of the field potential was significantly decreased and the latency period prolonged in group M and MN, but there was no significant difference in the above indices between group N and C. Compared with group M, the amplitude of the field potential was significantly increased and the latency period shortened in group MN. Compared with those 30 min before the tetanic stimulation, the amplitude of the field potential was significantly increased and latency period shorted at the time points after the tetanie stimulation in group C and N, the amplitude of the field potential was significantly decreased and latency period prolonged at the time points after the tetanie stimulation in group M, and the amplitude of the field potential was significantly increased 0-30 and 35-60 min after the tetanic stimulation and latency period shortened 0-30 min after the tetanie stimulation, the amplitude of the field potential was significantly decreased and latency period prolonged 65-120 and 125-210 min after the tetanic stimulation in group MN. Conclusion Morphine can inhibit synaptic LTP in the spinal dorsal horn evoked by electric stimulation of sciatic nerve in rats, and it may be one of the mechanisms of the central sensitization inhibition.
4.Value of combination of three-vessel views in ultrasound diagnosis of fetal aortic arch and pulmonary arterial branch anomalies
Yan XU ; Lianli JIE ; Zhirong JIANG ; Xiang GAO ; Jian CHENG
Chinese Journal of Ultrasonography 2015;(5):398-401
Objective To evaluate the role of combination of three-vessel views in ultrasound diagnosis of fetal aortic arch and pulmonary arterial branch anomalies.Methods The data of 66 cases of fetal aortic arch and pulmonary arterial branch anomalies were retrospectively analysed,including echocardiographic data,autopsy and operation records and postnatal follow-up results.Echocardiogaphic features and display frequencies on three vessels and trachea view(3VT),three vessels view(3VV)and three vessels and pulmonary arterial branches view(3VP)were summarized.Results There were 52 cases of aortic arch abnormalities,including 32 cases of right aortic arch with left ductus arteriosus,4 cases of double aortic arch,7 cases of aberrant right subclavian artery,6 cases of coarctation of aorta and 3 cases of interruption of aortic arch.Fourteen cases were diagnosed pulmonary aterial branch abnormalities,including 1 0 cases of crossed pulmonary arteries,2 cases of anomalous origin of left pulmonary artery from aortic arch and 2 cases of pulmonary artery sling.The display frequencies of fetal aortic arch and pulmonary arterial branch anomalies on 3VT,3VV and 3VP were 80.3%,19.7% and 39.4%.Conclusions The combination of three-vessel views is of great value in prenatal diagnosis of fetal aortic arch and pulmonary arterial branch anomalies.
5.Study on melatonin in suppression osteoblast proliferation and reducing the ratio of OPG/RANKL expression
Peng JIANG ; Jie JIANG ; Jiabo WANG ; Chao GAO ; Haibo FENG ; Yedong CHENG
Chinese Journal of Primary Medicine and Pharmacy 2016;23(15):2241-2243
Objective To investigate the effect of melatonin on osteoblast proliferation and osteogenic poten-tial in vitro cultured.Methods In vitro cultured human osteoblasts by different concentrations of melatonin (0, 1.0E -7,1.0E -6,1.0E -5,1.0E -4,1.0E -3mol/L)intervention 24 h,48 h,72 h,96 h.MTT assay was meas-ured by changes in cell proliferation,intervention in 96 hours,melatonin was measured by RT -PCR in cells OPG, RANK,RANKL influence,PTHLH mRNA expression.Results Compared with other groups,OD490 of the control group had statistical significance(all P <0.05).Compared with the control group,there were statistically significant differences in the ratio of OPG/RANKL of the dosing groups(all P <0.05).Conclusion Melatonin can promote the growth of bone into the cells and inhibit the action of bone cells into bone.
6.Effect of dominant accessory atrioventricular pathways on QRS complex terminal vector
Zhenfeng WU ; Jie HUI ; Xujie CHENG ; Bin JIANG ; Tingbo JIANG ; Zhihua LIU ; Jianping SONG ; Xiangjun YANG ; Wenping JIANG
Clinical Medicine of China 2008;24(8):785-787
Objective To analyze the effect of dominant accessory atrioventricular pathways (AP) on the end vector of ventricular depolarization. Methods All patients had single AP confirmed by radiofrequency cathteter abalation (RFCA) and were free from organic heart disease (including 102 cases of dominant accessory AP and 38 cases of concealed AP). The AP was divided into posterior septal(P3) ,mediate septal (MS) ,anterior septal (AS), left posterior free wall (LP), left anterior free wall (LA), right posterior free wall (RP) and right anterior free wall (RA). Results The end 40 ms vector of QRS wave changed in 102 patients with manifested AP and in 4 patients with concealed AP (P < 0. 05). Conclusion The end 40 ms vector of QRS wave of any site manifested AP can change and the changes have the specihty of leads.
7.The pathogen surveillance of rubella virus in Beijing from 2007 to 2010
Tiegang ZHANG ; Jie YANG ; Meng CHEN ; Cheng GONG ; Ming LUO ; Jiang WU ; Yingchun HUANG
Chinese Journal of Microbiology and Immunology 2011;31(5):403-405
Objective To clarify the pathogen for rubella in Beijing from 2007 to 2010. Methods Beijing Center for Disease Preventipn and Control ( CDC ) collected the specimens (including blood, urine and throat swab specimens) frqm clinically diagnosed rubella cases for serological test and virus isolation. The nucleic acid of rubella virus in clinical specimens and isolations was detected by real-time PCR. Results Fifty-five out of 99 blood specimens were positive for anti- rubella IgM. Fifty-one out of 99 clinically diagnosed rubella cases were confirmed as rubella cases by virus isolation. Seventy-two were confirmed as rubella virus infections with real-time PCR method for detecting the nucleic acid of rubella virus in clinical specimens. Compared with the sequences of reference strains of rubella virus, all of detected rubella virus belonged the IE gene type. Conclusion This study indicates that IE gene type virus was the predominant endemic rubella virus in Beijing.
8.Establishment of ischemic precondition model and the protective effect of nitric oxide on PC12 cell line
Lei JIANG ; Jun WANG ; Bin HE ; Fei HUAN ; Jie CHENG ; Rong CAO ; Hang XIAO ; Jinsong ZHANG
Chinese Journal of Emergency Medicine 2009;18(5):475-478
Objective To establish the ischemic precondition ([PC) model of PC12 cell line in vitro, and to explore the effect of nitric oxide (NO) on the IPC cerebral protection. Method PC12 cells were cultured and used for producing the model of ischemie precondition by the way of oxygen-glucose deprivation. Twenty dishes of cells were randomly divided into four groups (5 dishes for each group): control group, ischemic precondition group (IPC),non-ischemic precondition group (NIPC) and L-NAME treatment group (L-NAME). In control group, the cells were in-cubated with low glucose (<1 g/L) and2% FBS medium in normal oxygen; in IPC group, the cells were administrated with oxygen-glucose deprivation (OGD) for 6 hours, and then subjected with reperfuaion before OGD 15 hours; in NIPC group, the cells were treated the same as control group for 6 hours, and then subjected with reperfusion before OGD 15 hours; in L-NAME group, the cells received L-NAME (1 mmol/L) and cocultured for 30 minutes before OGD 6 hours, and then received the same treatment as the IPC group. To test whether the model was established, metabolic rate of MIT, LDH release were measured and the apoptosis rate was detected by flow cytometry following oxygen-glucose deprivation 15 hours. The activity of nitric oxide synthases (NOS) was as-sessed by biochemical assay. One-way ANOVA and LSD multiple comparison test were used to analyze differences among different groups, and P<0.05 was considered different. Results Compared with NIPC group, the metabolic rate of MTT increased (94.9%±35.1%, P<0.05), while LDH release and the cell apoptotic rate decreased significantly in IPC group (279.1%±28.1%, P<0.03). Compared with control group(100.0%± 13.5%),the activities of NOS increased both in NIPC and IPC groups (390.0%±14.6%, P<0.01;126.3% ±10.6%, P<0.01). Moreover, the apoptosis rates in each group (control group, IPC group, NIPC group and L-NAME group) were 5.90, 8.73, 38.62 and 11.73%,respectively. Conclusions IPC reduces the death and apoptosis rate of PC12 cell after oxygen-glucose deprivation injury. NO might be involved, but it is not the only factor.
9.Differental long non-coding RNA expression profiles in mice kidney during aging
Jie LI ; Fanfan GAO ; Lei CHEN ; Limin WEI ; Ning QU ; Xin CHENG ; Hongli JIANG
Chinese Journal of Nephrology 2021;37(3):209-216
Objective:To analyze the differentally expressed long non-coding RNA (lncRNA) among mice of different ages and explore the mechanism of kidney aging.Methods:Male C57BL/6 mice aged 3-month-old ( n=5), 12-month-old ( n=5) and 24-month-old ( n=5) (each weighting about 25 g) were randomly selected. PAS staining, Masson staining and senescence associated β-galactosidase (SA-β-gal) staining were used to detect the pathology and cell senescence of mice kidney. High throughput sequencing was performed to detect the differentially expressed lncRNA and their fragments per kilobase million. Real-time quantitative PCR was used to verify the differentially expressed lncRNA. Competitive endogenous RNA (ceRNA) network, which consisted of lncRNA, miRNA and mRNA was built. GO and KEGG enrichment analysis method were used to predict the biological function of differentially expressed lncRNA. Results:PAS staining and Masson staining showed the development of kidney fibrosis, and SA-β-gal staining positive region was increased significantly as age increased. There were 938 known lncRNA and 542 novel lncRNA differentially expressed among different ages' mouse kidney. Compared with 3-month-old mice, 33 lncRNA were up-regulated and 43 lncRNA were down-regulated in 12-month-old mice. Compared with 3-month-old mice, 130 lncRNA were up-regulated and 91 lncRNA were down-regulated in 24-month-old mice. Compared with 12-month-old mice, 36 lncRNA were up-regulated and 22 lncRNA were down-regulated in 24-month-old mice. The results of qRT-PCR about verified 10 lncRNAs with larger differential expression multiples and higer expression levels were consistent with the sequencing data. GO enrichment analysis showed that the target genes of lncRNA differentially expressed in the three groups were mostly located in the nucleus and cytoplasm, and might play a role by binding to proteins or participate in various protein phosphorylation, cell cycle, transcription, transcription regulation and other processes. KEGG enrichment analysis showed that the target genes of lncRNA differentially expressed in the three group were significantly enriched in Rap1 signaling pathway, FOXO signaling pathway and MAPK signaling pathway, which were closely related to kidney aging.Conclusion:There are significant differences in expression of lncRNA among the kidney of different ages mice, which are involved in the occurrence of renal senescence.
10.Simultaneous Determination of 12 Kinds of Organophosphates inWater and Sediment by High Performance LiquidChromatography-Tandem Mass Spectrometry
Meihong CHEN ; Huaizhou XU ; Ninghui SONG ; Shengmin WU ; Jie CHENG ; Jiang LI ; Shenghu ZHANG ; Lili SHI
Chinese Journal of Analytical Chemistry 2017;45(7):987-995
A method was developed for determination of 12 kinds of phosphate compounds in water and sediment by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) coupled with solid phase extraction (SPE) and ultrasonic extraction.The water samples were concentrated by HLB solid-phase extraction (SPE) column and eluted twice with ethyl acetate, ultrasonic solvent extraction for sediment samples and then repeated the operation of water samples after diluted with deionized water.The sample were separated on a ZORBAX Eclipse Plus C18 (150 mm × 2.1 mm, 3.5 μm) column by a gradient elution with 0.2% formic acid aqueous solution and methanol as the mobile phase.Ion mode analysis was monitored by high performance liquid chromatography mass spectrometer (MRM).The target compounds were quantified by external standard method.At the spiked levels (0.05, 0.1 and 0.5 μg/L), the average recoveries of 12 kinds of OPEs in water samples ranged from 66.4% to 115%, except for TMP (28.5%-47.8%) and TEHP (22.4%-73.8%).The relative standard deviation RSD (n=3) was 0.5%-9.09%, and the method quantification (MOQ) was 0.001-0.05 μg/L, However at the spiked levels of 5, 10 and 50 μg/kg, the average recoveries of 12 kinds of OPEs in sediment samples ranged from 65.4% to 120.0%, except for TMP (35.7%-44.9%) and TCEP (31.2%-48.9%).The relative standard deviation RSD (n=3) was 0.01%-9.54%, and the MOQ for sediment was 0.02-2.0 μg/kg dw.Based on the above methods, the detection and analysis of the targets in the water and sediments samples of Taihu Lake were carried out.The results showed that the concentrations of ΣOPEs were 0.1-1.7 μg/L and 8.1-420 μg/(kg dw), respectively.