The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

BACKGROUND:Neurodegenerative diseases are closely related to the imbalance of mitochondrial autophagy regulation.Previous studies by the research group have shown that lycium barbarum polysaccharide has neuroprotective effects,but whether it can improve the damage of SH-SY5Y cells induced byβ-amyloid protein 1-42 by regulating mitochondrial autophagy is still unclear.OBJECTIVE:To explore the protective effect and mechanism of Lycium barbarum polysaccharide on SH-SY5Y cells induced by β-amyloid protein 1-42.METHODS:An Alzheimer's disease cell model was established by inducing SH-SY5Y cells with β-amyloid protein 1-42,and then intervening with Lycium barbarum polysaccharide.SH-SY5Y cells were divided into three groups:control group,β-amyloid protein 1-42 group(20 μmol/L β-amyloid protein 1-42 for 24 hours),and Lycium barbarum polysaccharide group(1 g/L Lycium barbarum polysaccharide was added 1 hour in advance to form a protective effect,and then 20 μmol/L β-amyloid protein 1-42 was added to intervene with Lycium barbarum polysaccharide for 24 hours).CCK8 assay was used to detect cell viability.Mitochondrial membrane potential was detected by JC-1.TUNEL staining was used to detect cell apoptosis.Immunofluorescence and western blot assay were used to detect the expression of synaptic,apoptosis,and mitophagy-related indicators.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability of the β-amyloid protein 1-42 group decreased(P＜0.05);cell apoptosis rate increased(P＜0.05);mitochondrial membrane potential decreased(P＜0.05);the expressions of pro-apoptotic proteins Bax and Caspase3 increased(P＜0.05);the expression of anti-apoptotic protein Bcl-2 decreased(P＜0.05);the expression levels of synaptic-related proteins Syn and PSD-95 decreased(P＜0.05);the expression levels of mitochondrial autophagy-related proteins Pink1,LC3A/B,Parkin,and Beclin-1 decreased(P＜0.05);and the expression of P62 increased(P＜0.05).(2)Compared with the β-amyloid protein 1-42 group,the cell viability in the Lycium barbarum polysaccharide group was increased(P＜0.05);the apoptosis rate was decreased(P＜0.05);the mitochondrial membrane potential was increased(P＜0.05);the expression levels of Bax and Caspase3 were decreased(P＜0.05);the expression of Bcl-2 was increased(P＜0.05);the expressions of Syn and PSD-95 were increased(P＜0.05);the expression levels of Pink1,LC3A/B,Parkin,and Beclin-1 were increased(P＜0.05),and the expression of P62 was decreased(P＜0.05).These findings indicate that Lycium barbarum polysaccharide may inhibit β-amyloid protein 1-42-induced damage to SH-SY5Y cells by regulating mitophagy,reduce cell apoptosis,and increase neuronal synaptic plasticity.

Objective:To investigate the neuroprotective effect of nicotinamide riboside(NR)on experimental autoimmune en-cephalomyelitis(EAE)mice.Methods:C57BL/6 female mice were induced by myelin oligodendrocyte glycoprotein(MOG35-55)to pro-duce EAE model and were randomly divided into EAE group and NR group.From day 3 to day 27 after immunization,each mouse in EAE group was given normal saline(200 μl/d)and each mouse in NR group was given NR(500 mg/kg,200 μl/d)by intragastric administration.Clinical score and body weight of mice in EAE group and NR group were recorded every day.On the 28th day after immunization,the spinal cord protein of mice in each group was extracted and the frozen sections of spinal cord of mice in each group were prepared.LFB staining was used to detect demyelination,immunofluorescence staining was used to detect the expression of MAP-2 and the number of positive cells of NeuN,BDNF,GDNF,NGF and NT-3,and Western blot was used to detect the expressions of BDNF,GDNF,NGF and NT-3 of spinal cord.Results:Compared with EAE group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05),reduced weight loss,alleviated spinal cord demyelination(P<0.05),increased the expression of MAP-2(P<0.01)and the number of NeuN positive cells(P<0.01),and up-regulated the expressions of BDNF,GDNF,NGF and NT-3(P<0.05).Conclusion:NR shows a good neuroprotective effect on EAE mice.The mechanism may be related to NR significantly increasing the expression of spinal neurotrophic factors,improving the microenvironment of the central nervous sys-tem,nourishing nerves,promoting nerve repair and nerve growth,etc.

The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

BACKGROUND:Neurodegenerative diseases are closely related to the imbalance of mitochondrial autophagy regulation.Previous studies by the research group have shown that lycium barbarum polysaccharide has neuroprotective effects,but whether it can improve the damage of SH-SY5Y cells induced byβ-amyloid protein 1-42 by regulating mitochondrial autophagy is still unclear.OBJECTIVE:To explore the protective effect and mechanism of Lycium barbarum polysaccharide on SH-SY5Y cells induced by β-amyloid protein 1-42.METHODS:An Alzheimer's disease cell model was established by inducing SH-SY5Y cells with β-amyloid protein 1-42,and then intervening with Lycium barbarum polysaccharide.SH-SY5Y cells were divided into three groups:control group,β-amyloid protein 1-42 group(20 μmol/L β-amyloid protein 1-42 for 24 hours),and Lycium barbarum polysaccharide group(1 g/L Lycium barbarum polysaccharide was added 1 hour in advance to form a protective effect,and then 20 μmol/L β-amyloid protein 1-42 was added to intervene with Lycium barbarum polysaccharide for 24 hours).CCK8 assay was used to detect cell viability.Mitochondrial membrane potential was detected by JC-1.TUNEL staining was used to detect cell apoptosis.Immunofluorescence and western blot assay were used to detect the expression of synaptic,apoptosis,and mitophagy-related indicators.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability of the β-amyloid protein 1-42 group decreased(P＜0.05);cell apoptosis rate increased(P＜0.05);mitochondrial membrane potential decreased(P＜0.05);the expressions of pro-apoptotic proteins Bax and Caspase3 increased(P＜0.05);the expression of anti-apoptotic protein Bcl-2 decreased(P＜0.05);the expression levels of synaptic-related proteins Syn and PSD-95 decreased(P＜0.05);the expression levels of mitochondrial autophagy-related proteins Pink1,LC3A/B,Parkin,and Beclin-1 decreased(P＜0.05);and the expression of P62 increased(P＜0.05).(2)Compared with the β-amyloid protein 1-42 group,the cell viability in the Lycium barbarum polysaccharide group was increased(P＜0.05);the apoptosis rate was decreased(P＜0.05);the mitochondrial membrane potential was increased(P＜0.05);the expression levels of Bax and Caspase3 were decreased(P＜0.05);the expression of Bcl-2 was increased(P＜0.05);the expressions of Syn and PSD-95 were increased(P＜0.05);the expression levels of Pink1,LC3A/B,Parkin,and Beclin-1 were increased(P＜0.05),and the expression of P62 was decreased(P＜0.05).These findings indicate that Lycium barbarum polysaccharide may inhibit β-amyloid protein 1-42-induced damage to SH-SY5Y cells by regulating mitophagy,reduce cell apoptosis,and increase neuronal synaptic plasticity.

Objective:To investigate the neuroprotective effect of nicotinamide riboside(NR)on experimental autoimmune en-cephalomyelitis(EAE)mice.Methods:C57BL/6 female mice were induced by myelin oligodendrocyte glycoprotein(MOG35-55)to pro-duce EAE model and were randomly divided into EAE group and NR group.From day 3 to day 27 after immunization,each mouse in EAE group was given normal saline(200 μl/d)and each mouse in NR group was given NR(500 mg/kg,200 μl/d)by intragastric administration.Clinical score and body weight of mice in EAE group and NR group were recorded every day.On the 28th day after immunization,the spinal cord protein of mice in each group was extracted and the frozen sections of spinal cord of mice in each group were prepared.LFB staining was used to detect demyelination,immunofluorescence staining was used to detect the expression of MAP-2 and the number of positive cells of NeuN,BDNF,GDNF,NGF and NT-3,and Western blot was used to detect the expressions of BDNF,GDNF,NGF and NT-3 of spinal cord.Results:Compared with EAE group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05),reduced weight loss,alleviated spinal cord demyelination(P<0.05),increased the expression of MAP-2(P<0.01)and the number of NeuN positive cells(P<0.01),and up-regulated the expressions of BDNF,GDNF,NGF and NT-3(P<0.05).Conclusion:NR shows a good neuroprotective effect on EAE mice.The mechanism may be related to NR significantly increasing the expression of spinal neurotrophic factors,improving the microenvironment of the central nervous sys-tem,nourishing nerves,promoting nerve repair and nerve growth,etc.

Objective To observe the relationship between functional connectivity(FC)of frontoparietal network(FPN)and cognitive function in patients with cerebral small vessel disease(CSVD)using resting-state functional MRI(rs-fMRI).Methods rs-fMRI of 50 CSVD patients with cognitive impairment(CI group),65 CSVD patients with normal cognition(NC group)and 60 healthy controls(HC group),as well as outcomes of neuropsychological tests were retrospectively analyzed.Brain regions with different FC of FPN were compared among 3 groups and between each 2 groups.Partial correlation analysis was used to evaluate the correlations of FC of brain regions value being statistically different between CI and NC groups and cognitive scores.Results Significant differences of FC in bilateral cingulate gyrus,left middle frontal gyrus,right supramarginal gyrus,right inferior parietal lobule and right medial superior frontal gyrus were found among groups(FWE correction,all P＜0.05).Compared with NC group,FC of left cingulate gyrus decreased,of right inferior frontal gyrus and right medial superior frontal gyrus increased in CI group(FWE correction,all P＜0.05).The decreased FC value of left cingulate gyrus was negatively correlated with clock drawing test score in CSVD patients(r=-0.159,P=0.049).Conclusion CSVD patients with or without CI had extensive abnormal FC of FPN,and the left cingulate gyrus was associated with patient's cognitive function.

Objective To explore the relationship between serum heme oxygenase-1(HO-1),CD39 levels and cardiac function and major adverse cardiovascular events(MACE)in patients with successful cardiopulmo-nary resuscitation.Methods A total of 160 patients admitted to the Zigong First People's Hospital from De-cember 2019 to December 2022 who underwent successful cardiopulmonary resuscitation due to sudden heart-beat and respiratory arrest were selected as the study group.The study group was divided into cardiac function grade Ⅱ group,cardiac function grade Ⅲ group and cardiac function grade Ⅳ group using the New York Heart Association(NYHA)cardiac function grading method.In addition,100 healthy subjects who underwent physical examination in the same period were selected as the control group.The levels of serum HO-1 and CD39 in two groups were detected.After 6 months of follow-up,there were 91 cases in non-MACE group and 69 cases in MACE group.The predictive value of serum HO-1 and CD39 levels for MACE in patients with successful cardiopulmonary resuscitation was evaluated by receiver operating characteristic curve.Binary Lo-gistic stepwise regression analysis was used to investigate the influencing factors of MACE in patients with successful cardiopulmonary resuscitation.Results The serum levels of HO-1 and CD39 in the study group were higher than those in the control group,and the difference was statistically significant(P＜0.05).The levels of serum HO-1 and CD39 in cardiac function grade Ⅳ group were higher than those in cardiac function grade Ⅲ group and cardiac function grade Ⅱ group,and the difference was statistically significant(P＜0.05).Spearman correlation analysis showed that serum HO-1 and CD39 levels were positively correlated with NY-HA cardiac function classification in patients with successful cardiopulmonary resuscitation(r=0.534,0.496,P＜0.001).The serum levels of HO-1 and CD39 in MACE group were higher than those in non-MACE group,and the difference was statistically significant(P＜0.05).The area under the curve(AUC)of serum HO-1 and CD39 to predict MACE in patients with successful CPR were 0.725(95％CI:0.682-0.774)and 0.864(95％CI:0.814-0.917),respectively,and the AUC of the combined prediction was 0.901(95％CI:0.851-0.946).The time from cardiac arrest to resuscitation in non-MACE group was lower than that in MACE group,and the left ventricular ejection fraction(LVEF)was higher than that in MACE group,the difference was statistically significant(P＜0.05).Binary Logistic stepwise regression analysis showed that LVEF＜50％(OR=3.466,95％CI:1.537-7.818),serum HO-1≥5.13 ng/mL(OR=3.804,95％CI:1.706-8.479)and serum CD39≥29.87％(OR=4.345,95％CI:1.852-10.192)were the risk factors for MACE in patients with successful cardiopulmonary resuscitation(P＜0.05).Conclusion Serum HO-1 and CD39 levels have high predictive value for MACE in patients with successful cardiopulmonary resuscitation.

Objective To explore the differences between baduanjin and ninth set of broadcast gymnastics in their own pulse parameters before and after intervention for 12 weeks,and to provide objective indicators for the function of Baduanjin from the perspective of pulse diagnosis.Methods 60 healthy subjects were recruited,and 50 subjects were finally included according to the dropout criteria.The subjects were divided into the experimental group and the control group by the random number table method,and different interventions were adopted.Then,the pulse diagnosis of the two groups of subjects was collected,and the impact of the interventions on the viscera was judged by analyzing the Pulse Graph Parameters and contribution.Results The characteristic parameters of the experimental group include 8 time domain parameters(left Chi,left Guan,right Cun,etc.)and 9 frequency domain parameters(left Guan,left Chi,right Guan,etc.),which are mainly distributed in the left Cun,left Guan,left Chi,and right Guan;the characteristic parameters of the control group included 6 time domain parameters(left Cun,left Guan,right Chi,etc.)and 11 frequency parameters(left Guan,left Chi,right Guan,etc.),which were mainly distributed in left Chi,right Chi,and left Guan.Conclusion Practicing Baduanjin can significantly affect pulse diagram,and the accuracy and specificity after intervention are higher than those of the control group.Baduanjin has a certain influence on the circulation of qi and blood in the human meridians,and its mechanism of action may be related to affecting the overall circulation of qi and blood.In the experimental group,Baduanjin had a significant effect on the heart,liver,gallbladder,and kidneys,while the control group had the most obvious changes in the kidneys.For the overall intervention and coordination of the viscera,the Baduanjin group had better effects than the ninth broadcast gymnastics group.

ObjectiveTo study the effect of anodal transcranial direct current stimulation (atDCS) combined with mirror therapy (MT) on upper limb function for stroke patients. MethodsFrom July, 2018 to June, 2019, 54 stroke patients from the Department of Rehabilitation Medicine, Xuanwu Hospital, Capital Medical University, were randomly divided into groups A, B and C, with 18 cases in each group. All the patients received routine medication and rehabilitation training, while group A accepted 60 minutes of MT, group B accepted 20 minuts of atDCS first and then 40 minuts of MT, and group C accepted 40 minutes of MT first and then 20 minuts of atDCS in addittion; for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Action Research Arm Test (ARAT), Motor Assessment Scale (MAS) and modified Barthel Index (MBI) before and after treatment. ResultsThe main effect of time was significant in scores of FMA-UE (F = 159.811, P < 0.001), ARAT (F = 353.227, P < 0.001), MAS (F = 513.494, P < 0.001) and MBI (F = 85.982, P < 0.001), and all of them improved after treatment. The main effect of groups was significant in scores of FMA-UE (F = 12.502, P < 0.001), ARAT (F = 20.095, P < 0.001), MAS (F = 16.371, P < 0.001) and MBI (F = 11.882, P < 0.001). The interacted-effect between time and groups was significant in scores of FMA-UE (F = 37.659, P < 0.001), ARAT (F = 78.681, P < 0.001), MAS (F = 97.997, P < 0.001) and MBI (F = 48.015, P < 0.001); while all the scores were the best in group B (P < 0.01), and there was no significant difference between groups A and C (P > 0.05). ConclusionThe combination of atDCS first and then MT can promote the recovery of motor function of the affected limbs for stroke patients.