1.Effects of chronic ethanol consumption on field potential of cerebellar molecular layer in mice and the NO signal mechanism
Guanghui DONG ; Wenjing LI ; Liangyan LIU ; Chengquan LIN ; Songbiao CUI
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(5):391-396
Objective:To investigate the effect of chronic ethanol consumption on sensory information transmission in the cerebellar molecular layer and reveal the mechanism of chronic alcoholism on sensory information transmission and integration in the cerebellar cortex.Methods:Fifty healthy male ICR mice aged 6-8 weeks were randomly divided into saline group(control group)and ethanol consumption group(alcohol group) according to the random number table, with 25 mice in each group.The mice in alcohol group were injected intraperitoneally with 20% ethanol daily, while the mice in control group were injected with the same dose of normal saline. All mice were injected intraperitoneally once a day for 28 days.Through electrophysiological technology, patch-clamp amplifier and data acquisition software were used to record the changes in cerebellar molecular layer field potential of mice in the alcohol group and control group induced by sensory stimulation.Clampfit 10.3 software was used to record and analyze the electrophysiological data. SPSS 22.0 software was used for statistical analysis. Paired t-test and one-way ANOVA were used to analyze the differences before and after treatment. Results:After giving the stimulation of wind blowing, the amplitude of P1 in alcohol group was significantly higher than that in control group ((121.31±3.5)%, (97.2±2.7)%; t=26.08, P<0.05), and the area under the P1 curve (AUC) of the alcohol group was significantly lower than that of the control group ((127.1±4.2)%, (102.2±3.5)%; t=22.95, P<0.05). There was no significant difference in N1 amplitude between the two groups (P>0.05). When L-NNA, an inhibitor of nitric oxide synthase, was perfused into the brain surface of mice, the amplitude of P1 in alcohol group was significantly lower than that before administration ((76.2±4.8)%, (103.5±3.6)%; t=22.60, P<0.05), but there was no difference of the amplitude of P1 before administration and after elution ((101.5±4.6)%) ( t=1.70, P>0.05). After the L-NNA was perfused, the AUC of P1 was significantly lower than that before administration((72.4±5.6)%, (102.7±2.66)% ( t=24. 58, P<0.05), and there was no significant difference between before administration and after elution( (100.6±3.5)%, t=1.81, P>0.05). When L-NNA was perfused into the brain surface of mice, the amplitude of P1 in control group was (104.3±1.6)% and it had no differences compared with before administration(102.2±5.6)%, t=1.84, P>0.05) and after elution(102.5±4.5)%, t=1.92, P>0.05). And the AUC of P1 in control group after perfused L-NNA had no differences compared with before administration(103.5±2.6)%, (102.5±4.6)%) and after elution((101.9±3.7)%, t=0.99, 1.81, both P>0.05). When the mouse brain surface was perfused with NO donor SNAP, the amplitude of P1 in the control group was significantly higher than that before administration( (128.2±3.4)%, (103.5±2.6)%; t=28.89, P<0. 05) and there was no difference between before administration and after elution( (105.4±4.2)% , t=1.93, P>0.05). The AUC of P1((125.4±4.4)%) was higher than before administration((104.3±4.6)% , t=16.60, P<0.05) and there was no difference between before administration and after elution(103.5±4.2)%, t=0.65, P>0.05). Conclusion:Chronic ethanol consumption significantly enhances the inhibitory response, and the enhancement of inhibitory components stems from the activation of the NO signaling pathway.
2.Quantitative structure-activity relationship of synthetic pyrethroids and antibody activity
Xiujin CHEN ; Sihai HAN ; Chuanlai XU ; Jinying GUO ; Guoyan REN ; Daomin LI ; Songbiao LI ; Yuze HOU
Chinese Journal of Immunology 2015;(2):204-209
Objective:To research on the immune recognition mechanism of synthetic pyrethroids and generic specific antibody.Methods:We studied on quantitative structure-activity relationship ( QSAR ) of synthetic pyrethroids and their analogs as well as antibody activity ( IC50:fifty percent inhibition concentration ) using stepwise multiple linear regression method.Based on calculating structure descriptors of synthetic pyrethroids and their analogs , two-demensional QSAR ( 2D-QSAR ) model was established.The main factors affecting antibody activity were screened using 2D-QSAR,and predictive ability of QSAR models were evaluated by the method of leave-one-out( LOO) cross-validation.Meanwhile, the structure parameters of synthetic pyrethroid fragments were calculated and then analyzed using partial least squares ( PLS) assay.And then hologram QSAR ( H-QSAR) model was constructed on molecular substructure and antibody activity.The fragments contribution to antibody activity were illustrated by encoding different colors.Results:Decision coefficent (R2) of 2D-QSAR model and HQSAR model were 0.920 and 0.917 individually,cross-validation coefficient ( Q2 ) of two QSAR models were 0.875 and 0.660 respectively ,which showed two models had good predictive abil-ity.The result from 2D-QSAR model was also obtained that smaller was hydrophobicity of pyrethroids , easier was recognized by antibody.In addition,the optimum HQSAR model was constructed after we tried many combinations of these parameters .The fragment size in optimum HQSAR model was between 4 to 10,a hologram length was 61,optimum principle component was 4,and the fragment type of B/C/Ch was selected.However ,the fingerprint encoded results of synthetic pyrethroids weren′t consistent completely with exper-imental IC50 values.Conclusion:Hydrophobicity of synthetic pyrethroids is the largest correlation factors in antibody recognization .
3.Construction of Salmonella Pullorum C79-13ΔcrpΔasd mutant balanced-lethal host-vector system and its biological features
Songbiao CHEN ; Jing LI ; Ke SHANG ; Chuan YU ; Chunjie ZHANG ; Xiangchao CHENG ; Yinju LI ; Zhanqin ZHAO
Chinese Journal of Immunology 2014;(8):1083-1087,1092
In order to develop a safer vaccine strain exploit Salmonella Pullorum vaccine strain as a live vaccine vector.Methods:AΔcrpΔasd mutant of S.pullorum C79-13 strain was constructed and it was developed E.coli donor strain mutant was generated through the two-step method introduced by χ7213 ( pREΔasd) was conjugated with the recipient of C 79-13Δcrp.The C79-13ΔcrpΔasd the transduction of recombinant suicide plasmid .Results:PCR and sequencing results showed that ΔsipBSL1344 was suc-cessfully constructed.The further study indicated that foreign DAP must be supplied for the ΔcrpΔasd mutant to grow,in addition,the asd gene was transmitted stably .Compared with C79-13 strain,the O antigens was identical to C79-13 strain,but the growth velocity was reduced significantly ,significantly reduced virulence .Conclusion: The ΔcrpΔasd mutant can accept asd+plasmid to construct host-vector balance lethal system , and this system can be used to express foreign gene efficiently and to develop potential oral multivalent vaccines.
4.Paeoniflorin ameliorates neuropathic pain-induced depression-like behaviors in mice by inhibiting hippocampal neuroinflammation activated via TLR4/NF-kB pathway
Hualei BAI ; Shize CHEN ; Tiezheng YUAN ; Dongyuan XU ; Songbiao CUI ; Xiangdan LI
The Korean Journal of Physiology and Pharmacology 2021;25(3):217-225
Neuropathic pain (NP) that contributes to the comorbidity between pain and depression is a clinical dilemma. Neuroinflammatory responses are known to have potentially important roles in the initiation of NP and depressive mood. In this study, we aimed to investigate the effects of paeoniflorin (PF) on NP-induced depression-like behaviors by targeting the hippocampal neuroinflammation through the toll-like receptor 4 (TLR4)uclear factor-kappa B (NF-kB) signaling pathway. We used a murine model of NP caused by unilateral sciatic nerve cuffing (Cuff ). PF was injected intraperitoneally once a day for a total of 14 days. Pain and depression-like behavior changes were evaluated via behavioral tests. Pathological changes in the hippocampus of mice were observed by H&E staining. The levels of proinflammatory cytokines in the hippocampus were detected using ELISA. Activated microglia were measured by immunohistochemical staining. The TLR4/NF-kB signaling pathwayassociated protein expression in the hippocampus was detected by western blotting. We found that the PF could significantly alleviate Cuff-induced hyperalgesia and depressive behaviors, lessen the pathological damage to the hippocampal cell, reduce proinflammatory cytokines levels, and inhibit microglial over-activation. Furthermore, PF downregulated the expression levels of TLR4/NF-kB signaling pathwayrelated proteins in the hippocampus. These results indicate that PF is an effective drug for improving the comorbidity between NP and depression.
5.Paeoniflorin ameliorates neuropathic pain-induced depression-like behaviors in mice by inhibiting hippocampal neuroinflammation activated via TLR4/NF-kB pathway
Hualei BAI ; Shize CHEN ; Tiezheng YUAN ; Dongyuan XU ; Songbiao CUI ; Xiangdan LI
The Korean Journal of Physiology and Pharmacology 2021;25(3):217-225
Neuropathic pain (NP) that contributes to the comorbidity between pain and depression is a clinical dilemma. Neuroinflammatory responses are known to have potentially important roles in the initiation of NP and depressive mood. In this study, we aimed to investigate the effects of paeoniflorin (PF) on NP-induced depression-like behaviors by targeting the hippocampal neuroinflammation through the toll-like receptor 4 (TLR4)uclear factor-kappa B (NF-kB) signaling pathway. We used a murine model of NP caused by unilateral sciatic nerve cuffing (Cuff ). PF was injected intraperitoneally once a day for a total of 14 days. Pain and depression-like behavior changes were evaluated via behavioral tests. Pathological changes in the hippocampus of mice were observed by H&E staining. The levels of proinflammatory cytokines in the hippocampus were detected using ELISA. Activated microglia were measured by immunohistochemical staining. The TLR4/NF-kB signaling pathwayassociated protein expression in the hippocampus was detected by western blotting. We found that the PF could significantly alleviate Cuff-induced hyperalgesia and depressive behaviors, lessen the pathological damage to the hippocampal cell, reduce proinflammatory cytokines levels, and inhibit microglial over-activation. Furthermore, PF downregulated the expression levels of TLR4/NF-kB signaling pathwayrelated proteins in the hippocampus. These results indicate that PF is an effective drug for improving the comorbidity between NP and depression.
6.Construction and characterization of invasion protein B gene deleted mutant of Salmonella typhimurium SL1344 strain
Songbiao CHEN ; Jing LI ; Chuan YU ; Lei HE ; Chunjie ZHANG ; Xiangchao CHENG ; Yinju LI ; Yunfei YAN ; Xiuzhe JIN ; Guihua CHEN ; Zhanqin ZHAO
Chinese Journal of Immunology 2015;(2):215-220
Objective:In order to develop a safer vaccine strain exploit Salmonella typhimurium vaccine strain .A ΔsipB mutant of Salmonella typhimurium SL 1344 strain was constructed.Methods: Firstly, the recombinant suicide plasmid containing the missing 585 bp sipB ( PREΔsipB ) was built by homologous recombination , and screenned by two-step method.Results: PCR and sequencing results showed that SL 1344ΔsipB was successfully constructed.It was no significant changes compared with SL 1344.But compared with the parent strains SL 1344 , the mutant strain had obvious change in its virulence , oral challenge of bacteria in mice revealed that LD50 of the mutant strain was 1.70 ×108 CFU,the toxicity reduced about 1.4%.The protection rate induced by the sipB mutant was 50%,and the serum antibody peaked 14 d post-immunization.Conclusion:The SL1344ΔsipB mutant was constructed suc-cessfully,and genetic stability ,significantly reduced virulence.The study provides a new approach for further study of the relationship between the gene and pathogenicity of Salmonella typhimurium.It is likely that the ΔsipB mutant could be adapted to develope attenuated Salmonella vaccine.
7.Effects of long-term alcohol consumption on synaptic plasticity of mossy fiber-granule cells in cerebellar cortex and motor coordination function in mice
Guanghui DONG ; Wenjing LI ; Delai QIU ; Songbiao CUI
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(3):205-211
Objective:To investigate the long-term alcohol consumption on synaptic plasticity of mossy fiber-granule cells in cerebellar cortex and motor coordination function in mice.Methods:Thirty healthy male ICR mice aged 6-8 weeks were divided into saline group (control group)and alcohol consumption group(alcohol group) according to random number table with 15 in each group. The mice in alcohol group were injected intraperitoneally with 15% ethanol (1.6 g/kg), while the mice in control group were injected with the same volum of normal saline, all mice were injected intraperitoneally once a day for 28 consecutive days. Walking obstacle test and rotating rod fatigue test were used to observe the motor coordination ability and learning ability of mice. Electrophysiological patch clamp technique was used to detect the field potential changes of long-term synaptic plasticity induced by blowing stimulation. SPSS 22.0 software was used for statistical analysis.Independent sample t-test, paired t-test and repeated measurement analysis of variance were used for comparison between the two groups before and after intervention. Results:The electrophysiological results showed that the amplitude percentage of field potential N1 wave in the control group after blowing stimulation was (130.4±3.3)%, which was higher than that before stimulation ((100.6±2.7)%) ( t=27.07, P<0.01). And the percentage of area under N1 standardized waveform after stimulation ((128.8±4.5)%) was greater than that before stimulation ((100.2±3.5)%) ( t=19.43, P<0.01). There was no significant difference in the amplitude percentage of N1 wave in alcohol group ((97.8±4.3)%) after blowing stimulation compared with that before stimulation ((99.5±5.6)%) ( t=0.93, P>0.05). And also there was no significant difference in the area percentage under N1 wave after stimulation ((96.8±3.6)%) compared with that before stimulation ((100.2±4.2)%) ( t=2.38, P>0.05). The results of walking obstacle test showed that the total number of errors (3.14±0.19) in the alcohol group was higher than that in the control group(1.52±0.29) ( t=17.87, P<0.01), and the total error time ((63.85±9.34) ms) was longer than that in the control group ((28.93±7.21) ms) ( t=11.45, P<0.01). The results of repeated measurement analysis of variance showed that there was an interaction between time and group in the falling speed and falling latency of the two groups of mice in the rotating rod fatigue experiment ( F=4.5, 455.1, both P<0.05). The drop speed of mice in the alcohol group was significantly lower than that in the control group from day 1 to 7 (all P<0.05). The fall latency of mice in the alcohol group from day 1 to 7 was shorter than that in the control group, and the difference was statistically significant (all P<0.05). Conclusion:Long-term alcohol consumption impairs synaptic plasticity in the granular layer of mice and leads to a significant decline in motor coordination and motor learning ability.
8.The effects of urapidil on myocardial perfusion and cardiac function in acute myocardial infarction
Daokuo YAO ; Sanqing JIA ; Lei WANG ; Songbiao YAN ; Hongwei LI ; Yuchen ZHANG ; Hui CHEN ; Yongliang WANG ; Yupeng WANG ; Lin ZHAO ; Rongjing DING
Clinical Medicine of China 2008;24(9):932-935
Objective To evaluate the effect of urapidil on myocardial reperfusion and cardiac function during primary percutenons coronary intervention (PCI) in acute myocardial infarction (AMI) patients.Methods 54 AMI patients undergoing primary PCI were randomized to intracoronary urapidil (n=27) and nitroglycerin (n=27) group.When blood flow grade TIMI-l or more appeared in the infarct related artery (IRA) before or after percutenous transluminal coronary angioplasty (PTCA),urapidil (12.5 mg) or nitroglycerin (0.2 mg) was given intracoronary and then stents were implanted as needed.TIMI blood flow,no reflow/slow floW,corrected TIMl frame count (cTFC),myocardial blush grade (MBG),ST resolution (STR) on ECG, peak of creatine kinase (CK),cardiac troponin T (cTnT) were observed before and after PCI.Left ventricular ejection fraction (LVEF) was measured 24 hours and 30 days after PCI and MACE including death,reinfartion,revascularization in hospital were observed. Resuits Urapidil group compared to the nitroglycerin group,cTFC (18.38±3.30 vs.21.44±4.26,P=0.005) decreased and MBG (P=0.040) improved.STR in the urapidil group improved significantly compred to the nitroglycerin group (93% vs.70%,P=0.038).LVEF of 24 hours and 30 days after PCI in the urapidil group was higher than that ofthe nitroglycerin group (0.55±0.05 vs.0.52±0.06,P=0.021 and 0.58±0.06 vs 0.54±0.06,P=O.041,respectively).Peak CK (1895.26±1239.02 vs.1269.96±515.84,P=0.021) and peak TnT (5.81±5.27 vs 3.64±2.35,P=0.050) in the urapidil group decreased more significantly than that of the nitroglycerin group.No difference of MACE was found in the two groups.Conclusion Intracoronary urapidil administration adjunct to primary PCI in AMI ameliorates coronary blood flow and myocardial perfusion, improves ventricular function,and reduce the infarct size and does not incllase the incidence of complications during hospitalization.
9.Construction and characterization of invasion protein B gene deleted mutant asd host-vector balanced lethal system
Chunjie ZHANG ; Songbiao CHEN ; Xiangchao CHENG ; Chengshui LIAO ; Jing LI ; Lei HE ; Mingliang ZHANG ; Chuan YU ; Zuhua YU ; Yanyan JIA ; Zhanqin ZHAO
Chinese Journal of Immunology 2016;(2):210-213
Objective: To develop an oral live vaccine vector which stably carries exogenous genes.Methods:SL1344ΔsipBΔasd host-vector balanced lethal system was constructed by the method of recombinant suicide plasmid-mediated allelic exchange on the basis of attenuated Salmonella typhinurium SL1344ΔsipB.Then,the biological characteristics of SL1344ΔsipBΔasd was analyzed.Results:The results showed that the mutant was stabile with the Δasd gene in vitro;the serotype and growth rate of SL1344ΔsipBΔasd strain was almost same as the parent SL1344ΔsipB and SL1344 strain.And the mutant strains remain swim ming zones.Virulence test in mice showed that the virulence of SL1344ΔsipBΔasd which carried complementary plasmid pYA3493 by electro-transformation decreased by 1.4%compared with SL1344.Conclusion: These results showed that the SL1344ΔsipBΔasd mutant was successfully constructed.It is likely that this mutant should be used as a live vector to express foreign genes.