1.Clinical features and diagnosis in two cases general paresis of insane
Fuhua PENG ; Wei QIU ; Lianfang BIAN ; Xueqiang HU
Chinese Journal of Neurology 2001;0(02):-
Objective To analyse the clinical features in general paresis of insane(GPI)as to offering an early diagnosis. Methods A retrospective analysis of 2 cases of GPI was focused on their clinical manifestations. Results The clinical features of GPI showed: (1) Chronic onset, progressive development in 2 cases; (2) Dementia was key symptom.Hasegawa dementia scales were 4 and 17 in 2 cases.2 cases accompanied with abnormal mentality,such as delusion of grandeur ,euphoria and so on;(3)GPI have features of pupil change,dysarthria,muscular tension,abnormal reflection; (4) Serum and cerebrospinal fluid (CSF) syphilis antibody reaction manifested positive, CSF protein content was increased significantly, and cells increased ( with lymph cell mainly ); (5) Head MRI manifested: cerebral atrophy in 2 cases, multiple abnormal signals in cerebral parenchyma in 1 case. Conclusions GPI misdiagnosis rate is high. Important basis of diagnosis remains on clinical manifestations, laboratory and imaging examinations.
2.Cryopreservation of Cyclamen persicum Mill. Callus.
Fuhua BIAN ; Xueqin GONG ; Cuirong YOU ; Caixia ZHENG ; Funing QU
Chinese Journal of Biotechnology 2008;24(3):504-508
In this paper we studied cryopreservation of Cyclamen persicum Mill. callus to avoid variations produced by sub-culture. The callus in the logarithmic phase after sub-culture were used for experiments. Firstly, the callus were pre-cultured in culture-medium containing 4%, 6% or 8% sucrose for different time periods, transferred to different cryoprotectants to directly cryopreserve or incubated for 2 hours at -20 degrees C, then submersed in liquid nitrogen, lastly thawed rapidly in a waterbath at 37 degrees C, and washed with liquid culture-medium containing the corresponding concentration of sucrose. Cell survival rate was computed after stained by Neutral Red, and SPSS 13.0 software was used for statistical analysis. The results showed that sucrose concentration, pre-culture time, cryoprotectants had various impacts on cell survival rate. We have developed a simple but effective protocol for the cryopreservation of callus of C. persicum. Of the different protocols tested, 4%sucrose, pre-culturing for 3 days, No. 9 cryoprotectant and freezing directly after 30 minutes at 0 degrees C results in the highest cell survival rate.
Cryopreservation
;
Culture Techniques
;
methods
;
Cyclamen
;
growth & development
3.Systematic comparison of two kinds of Bufonis Venenum derived from different Bufo gargarizans subspecies based on metabolomics and antitumor activity.
Bo SUN ; Ming-Li LI ; Yao-Hua DING ; Yong ZHANG ; Bo XIA ; Sheng-Ya GUO ; Shi-Lu WANG ; Bao-Lin BIAN ; Nan SI ; Hai-Yu ZHAO
China Journal of Chinese Materia Medica 2023;48(5):1280-1288
This paper compared the differences between two kinds of Bufonis Venenum produced by Bufo gargarizans gargarizans and B. gararizans andrewsi, and verified the rationality of the market value orientation of Bufonis Venenum based on the zebrafish mo-del. Twenty batches of Bufonis Venenum from Jiangsu province, Hebei province, Liaoning province, Jilin province, and Liangshan, Sichuan province, including B. gargarizans gargarizans and B. gararizans andrewsi, were collected. The UHPLC-LTQ-Orbitrap-MS combined with principal component analysis was used to compare the differences between two kinds of Bufonis Venenum. According to the limiting conditions of VIP>1, FC<0.5 or FC>2.0, and peak total area ratio>1%, 9 differential markers were determined, which were cinobufagin, cinobufotalin, arenobufagin, resibufogenin, scillaredin A, resibufagin, 3-(N-suberoylargininyl)-arenobufagin, 3-(N-suberoylargininyl)-marinobufagin, and 3-(N-suberoylargininyl)-resibufogenin. The content of 20 batches of Bufonis Venenum was determined according to the Chinese Pharmacopoeia(2020 edition) by high-performance liquid chromatography, and the 2 batches of Bufonis Venenum, CS7(8.99% of total content) and CS9(5.03% of total content), with the largest difference in the total content of the three quality control indexes of the Chinese Pharmacopoeia(bufalin, cinobufagin, and resibufogenin) were selected to evaluate their anti-liver tumor activity based on the zebrafish model. The tumor inhibition rates of the 2 batches were 38.06% and 45.29%, respectively, proving that only using the quality control indexes of the Chinese Pharmacopoeia as the value orientation of Bufonis Venenum market circulation was unreasonable. This research provides data support for the effective utilization of Bufonis Venenum resources and the establishment of a rational quality evaluation system of Bufonis Venenum.
Animals
;
Zebrafish
;
Bufanolides/analysis*
;
Bufonidae
;
Chromatography, High Pressure Liquid
;
Quality Control
;
Cell Line, Tumor
4.Study on anti-inflammatory activity and mechanism of indolealkylamines in toad skin on LPS-activated neutrophils.
Yu ZHANG ; Yao-Hua DING ; Xiao-Lu WEI ; Yan-Yan ZHOU ; Nan SI ; Jian YANG ; Hong-Jie WANG ; Hai-Yu ZHAO ; Bao-Lin BIAN
China Journal of Chinese Materia Medica 2021;46(18):4774-4781
Indolealkylamines(IAAs) are the main hydrophilic substances in toad skin, mainly including free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine. In this study, the LPS-activated neutrophils were used to investigate the structure-activity relationship and anti-inflammatory mechanism of the above-mentioned five monomers from the toad skin in vitro. The neutrophils were divided into the control group, model group(1 μg·mL~(-1) LPS), positive drug group(100 μg·mL~(-1) indometacin), as well as the low-(50 μg·mL~(-1)), medium-(100 μg·mL~(-1)) and high-dose(200 μg·mL~(-1)) free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine groups. The levels of IL-6, TNF-α and IL-1β in the neutrophil supernatant of each group was measured by enzyme-linked immunosorbent assay(ELISA) after LPS stimulation, followed by the detection of apoptosis in each group after Annexin V/PI staining. The protein expression levels of caspase-3, Bax, Bcl-2, beclin1, LC3-I, and LC3-Ⅱ were assayed by Western blot. The results showed that IAAs reduced the excessive secretion of inflammatory cytokines caused by LPS compared with the model group. Besides, the activity of each free IAAs(N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine and dehydrobufotenine), especially bufotenine, was stronger than that of the binding bufothionine. As revealed by Annexin V/PI staining, LPS delayed the early apoptosis of neutrophils compared with the control group, while bufotenine promoted the apoptosis of neutrophils in a dose-dependent manner, which might be related to the elevated expression of apoptosis-related protein Bax/Bcl-2. In addition, LPS activated the autophagy pathways in neutrophils. This study confirmed the efficacy of IAAs in reducing the secretion of inflammatory cytokines in neutrophils induced by LPS for the first time. For instance, bufotenine exerts the anti-inflammatory effect possibly by inducing the apoptosis of neutrophils.
Animals
;
Anti-Inflammatory Agents/pharmacology*
;
Apoptosis
;
Bufonidae
;
Lipopolysaccharides/toxicity*
;
Neutrophils
;
Skin