1.Influence of pregnant rats' prenatal chronic stress on hippocampal dendritic spines and the function of learning and memory in their offsprings
Xicheng TAO ; De WU ; Jiulai TANG ; Ziyun ZHOU ; Jing ZHU ; Wencheng XU
Chinese Journal of Applied Clinical Pediatrics 2015;30(12):909-912
Objective To study the influence of pregnant rats' prenatal chronic stress (PS) on learning and memory of their offspring rats and its possible molecular mechanisms.Methods Pregnant females were individually restrained for 45 min 3 times a day during pregnancy from day 14 to day 21.Control pregnant females were left undisturbed in their home cages.The rat offsprings were randomly assigned to PS group or control group.Males and females were kept for the study separately.The learning and memory of the developing rat offspring in the Morris water maze were examined.The basal levels of corticosterone (COR) and adreno-cortico-tropic-hormone (ACTH) were analyzed by using radioimmunoassay.The Golgi-Cox impregnation technique was used to compare density and morphology of the CA1 hippocampal dendritic spines.Results The escape latency (EL) to find the platform in the control group was significantly less than that in the PS group in female rat offspring (F =4.533,P < 0.05),and the difference was statistically significant on the 5th day (t =2.788,P < 0.01).EL to find the platform in the control group was significantly less than that in the PS group in male rat offspring (F =6.101,P <0.05),and the difference was statistically significant on the second day (t =3.051,P < 0.01).In the space exploration experiments of the water maze,the retention time observed for the control group and the PS group in the goal quadrant was similar(P > 0.05).The basal levels of the serum COR in the PS group were higher than those in the control group of female rat offspring(t =3.658,P < 0.01) and the basal levels of the serum ACTH in the PS group were higher than those in the control group of male rat offsprings(t =2.319,P < 0.05).A simplified pattern was observed in the CA1 hippocampal dendritic spines in the PS group,showing a less extent of dendritic arborization and the density was significantly lower than that in the control group(t =-3.072,P < 0.01).Conclusions Altered function of the hypothalamic-pituitary-adrenal axis in the offspring mediates the cognitive alterations observed following prenatal stress should to be associated with the lower density and simplified pattern of CA1 dendritic spines.
2.To optimize dual molecular beacon detection system for rapid identification of Mycobacterium tuberculosis
Chuanxing YU ; Huan ZHANG ; Mingxiang HUANG ; Ziyun ZHAO ; Ling ZHU
International Journal of Laboratory Medicine 2018;39(9):1029-1033
Objective To optimize the experimental system of dual molecular beacon to rapidly detect My-cobacterium tuberculosis and its resistant strains.Methods Fluorescence quantitative PCR was carried out by selecting different magnesium ion concentration,annealing temperature and primer concentration respectively. Finally,the optimum reaction conditions were obtained.Results In order to ensure the efficiency of amplifica-tion and no non-specific amplification,the final selection of the best conditions were as follows,the concentra-tion of Mg2+was 3.0 mmol/L,annealing temperature was 60 ℃,and the concentration of primers was 0.3 mmol/L.Conclusion The optimal condition of dual molecular beacon experiment was established,which en-sured that the detection of Mycobacterium tuberculosis by molecular beacon quantitative PCR had the advanta-ges,such as simple operation,rapid speed,high sensitivity(the minimum detection limit was 1 CFU/mL)and specificity(only Mycobacterium tuberculosis complex including drug-resistant strains could be detected),good reproducibility(coefficient of variation was < 5%)and other advantages.The study provides the necessary conditions for the dual molecular beacon detection of Mycobacterium tuberculosis.
3.Short-term follow-up for unruptured wide-necked intracranial aneurysms treated with Pipeline embolization device
Xiang XIAO ; Guohua MAO ; Jianming ZHU ; Ziyun GAO ; Xianliang LAI ; Shuxin SONG ; Minhua YE ; Xingen ZHU
Chinese Journal of Cerebrovascular Diseases 2017;14(12):628-632,647
Objective To evaluate the short-term effect of Pipeline embolization device (PED)for the treatment of unruptured wide-necked intracranial aneurysms. Methods From October 2015 to September 2016,15 consecutive patients with unruptured wide-necked intracranial aneurysm (aneurysm neck and aneurysm body ratio ≥0. 5)treated with PED at the Department of Neurosurgery,the Second Affiliated Hospital of Nanchang University were enrolled retrospectively. Their clinical and imaging data were analyzed. Kamran scale was used to evaluate the embolization rate of aneurysms and the changes of the parent arteries. DSA examination was performed again at 6 -12 months after operation. Results Fifteen PED were implanted in 15 patients with unruptured wide-necked intracranial aneurysms,including 13ophthalmic artery aneurysms,1 posterior communicating artery aneurysm,and 1 cavernous sinus aneurysm. The technical success rate was 100% . Immediately after PED implantation,Karman rating of 15 cases were aneurysm grade 2 embolization,parent artery grade A (grade 2a). DSA examination was performed again at 6 - 12 months after operation showed that 14 patients were aneurysm grade 4,parent artery was grade A (grade 4a). One patient (ophthalmic artery aneurysm)underwent the second DSA examinations at 6 and 12 months after operation showed that the residual development of aneurysms. The aneurysm embolization was grade 3, and the parent artery was grade A (grade 3a). No branch artery occlusion was observed. Non of them had neurological deficit. The modified Rankin scale score was 0 in all 15 patients. Conclusion The use of PED in the treatment of unruptured wide-necked intracranial aneurysms has a higher occlusion rate. Its long-term effect still needs further follow-up.
4.Establishment of a high performance liquid chromatography method for the detection of CD38 enzyme activity in blood
Xianghui ZHAO ; Xiaoxiao BU ; Hanyu ZHU ; Hongxia LI ; Jihua SHI ; Siming WANG ; Hongna MU ; Ziyun LI ; Zhu WU ; Jun DONG ; Wenxiang CHEN ; Ruiyue YANG
Chinese Journal of Laboratory Medicine 2023;46(10):1074-1082
Objective:A high performance liquid chromatography (HPLC) method was developed to determine the enzymatic activity of CD38 in blood, which was the major enzyme responsible for consuming nicotinamide adenine dinucleotide (NAD). Additionally, the study aimed to detect the differences in CD38 enzymatic activity among individuals of varying ages and health statuses.Methods:A 50 μl whole blood matrix and enzyme reaction substrate of 150 μl β-NAD at a concentration of 500 μmol/L were selected for the analysis. To eliminate the impact of endogenous β-NAD, the whole blood sample was pre-incubated at 37 ℃ for 20 minutes before adding the substrate. The reaction was terminated by perchloric acid (PCA) after incubation at 37 ℃ for 40 min. The change in product nicotinamide (NAM) before and after the enzymatic reaction was measured by HPLC to calculate the CD38 activity. The linearity, limit of detection, limit of quantification, precision, and stability of the method were evaluated. The CD38 enzymatic activities in 60 healthy volunteers and 30 colorectal cancer patients in blood were determined by the developed method.Results:Pre-incubation at 37 ℃ for 20 minutes eliminated the effect of endogenous β-NAD. The correlation coefficient of NAM was 0.999 in the concentration range of 0.1-3.2 μmol/L, with limit of detection of 0.5 nmol/L and limit of quantification of 2.1 nmol/L. The average within-run imprecision ( CV) and total CV were 3.22%-4.03% and 2.91%-4.70%, respectively. The recovery rate ranged from 94.82% to 96.81%. The CD38 activity of whole blood was stable by storage at 4 ℃ for 48 hours, storage at room temperature for 8 hours, thawing of frozen whole blood at room temperature for 2 hours, or repeated freeze-thawing three times. NAM, NAD standards, and pre-treatment samples were stable after 48 hours at 4 ℃ and 8 hours at room temperature. CD38 activity gradually decreased with increasing concentration of the added CD38 inhibitor 4-aminoquinoline derivative (78c). Measurement of 60 healthy physical examination population samples showed significantly higher CD38 enzyme activity in the elderly group than that in the young group ( t=-2.776, P=0.007) and measurement of 30 colorectal cancer patients showed significantly higher CD38 enzyme activity than that in healthy people ( t=-2.572, P=0.012). Conclusion:The established HPLC method for determining CD38 enzymatic activity is characterized by its simplicity, efficiency, accuracy, and reproducibility. This technique serves as a valuable tool for investigating aging and aging-related diseases.
5.Recovery of proprioception after lateral ankle sprain
Renjie XU ; Zhou LI ; Yuting GUO ; Xiqin YU ; Jingming MA ; Xiangyang GE ; Ziyun ZHU ; Yuxin ZHANG ; Feng ZHOU
Chinese Journal of Rehabilitation Theory and Practice 2023;29(7):844-848
ObjectiveTo observe the recovery of proprioception of the affected ankle over time after lateral ankle sprain accepting routine rehabilitation. MethodsFrom June, 2020 to June, 2022, 18 patients with lateral ankle sprain in Kunshan Rehabilitation Hospital underwent routine rehabilitation for twelve weeks. They were measured active and passive position sense of bilateral ankles using an isokinetic dynamometer before treatment, and four, eight and twelve weeks after treatment, respectively. ResultsThe active presentation difference of affected ankle reduced after treatment (F = 22.533, P < 0.001), but it was more than that of the healthy ankle at the same time (t > 4.419, P < 0.001). No significant improvement was found in passive presentation difference of affected ankle after treatment (F = 1.175, P > 0.05), and it was not significantly different from those of the healthy ankle at the same time (|t| < 0.646, P > 0.05). ConclusionProprioception of affected ankle has been impaired after lateral ankle sprain, and it can be recovered after rehabilitation, but cannot achieve the healthy level even after three months of training. Passive position sense as an index of proprioception needs more researches.