1.Protective effect of transection of cervical sympathetic trunk on noise-induced hearing loss in guinea pigs
Xuefeng ZHANG ; Zhimin TAN ; Zongbao ZHANG ; Haitao WANG
The Journal of Practical Medicine 2015;31(23):3840-3842
Objective To investigate the protective effect of transection of cervical sympathetic trunk (TCST) on the noise-induced hearing loss (NIHL) in guinea pigs. Methods Thirty-six guinea pigs were ran-domly divided into three groups (n = 12): the operation group, the sham group and the blank control group. TCST was performed in the operation group , while the cervical sympathetic trunk in the sham group was only ex-posed and separated. The auditory brainstem response (ABR) thresholds was measured. Levels of superoxide dis-mutase (SOD) and malondialdehyde (MDA) in pasma were detected before operation (T0), before noise expo-sure (T1) and on the time of stopping noise exposure (T2), respectively. The damage situation of the outer cochlear hair cells was also checked. Results (1) At the time of T2, ABR thresholds of operation group and sham group were higher than that of the blank control group (P < 0.01), and ABR thresholds of the sham group are higher than the of the operation group (P < 0.05). (2) Levels of SOD and MDA post-noise exposure (T2) were significantly different among the three groups (P < 0.05). (3) Injured hair cells were more serious in the sham group than that in the operation group. Conclusion TCST has protective effect on NIHL in guinea pigs through increasing SOD and decreasing MDA.
2.Expression, purification and characterization of a novel fatty acid synthase from Rhodosporidium toruloides.
Zhiwei ZHU ; Sufang ZHANG ; Xinping LIN ; Wujun LIU ; Zongbao K ZHAO
Chinese Journal of Biotechnology 2014;30(9):1414-1423
Fatty acid synthase (FAS) catalyses the reaction between acetyl-CoA and malonyl-CoA to produce fatty acids. It is one of the most important enzyme in lipid biosynthesis. FAS of the oleaginous yeast Rhodosporidium toruloides has two acyl carrier protein (ACP) domains and a distinct subunit composition compared with FASs of other species. As ACP is a protein cofactor crucial for fatty acid chain elongation, more ACPs in the FAS may facilitate the reaction. To study the biochemical and structural properties of this novel FAS from R. toruloides, plasmids were constructed and transformed into Escherichia coli BL21 (DE3). The strain ZWE06 harboring plasmids pET22b-FAS1 and pET24b-FAS2 could co-overexpress the two subunits. The recombinant FAS was purified by sequentially using ammonium sulphate precipitation, sucrose density gradient centrifugation and anion exchange chromatography. The specific activity of the recombinant FAS was 548 mU/mg. The purified complex would be used to study enzyme kinetics and protein structure of FAS, and heterogeneous expression and purification will facilitate revealing the mechanism of this novel FAS with double ACPs.
Acyl Carrier Protein
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Basidiomycota
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enzymology
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Chromatography
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Escherichia coli
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metabolism
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Fatty Acid Synthases
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biosynthesis
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genetics
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Fatty Acids
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biosynthesis
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Plasmids
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Recombinant Proteins
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biosynthesis
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genetics
3.Cross Frequency Coupling Characteristic Analysis in Subthalamic Local Field Potentials of Parkinson's Disease.
Zongbao WANG ; Yongzhi HUANG ; Xinjing ZHANG ; Xinyi GENG ; Xiao CHEN ; Shouyan WANG
Journal of Biomedical Engineering 2015;32(4):874-880
Pathological neural activity in subthalamic nucleus (STN) is closely related to the symptoms of Parkinson' s disease. Local field potentials (LFPs) recordings from subthalamic nucleus show that power spectral peaks exist at tremor, double tremor and tripble tremor frequencies, respectively. The interaction between these components in the multi-frequency tremor may be related to the generation of tremor. To study the linear and nonlinear relationship between those components, we analyzed STN LFPs from 9 Parkinson's disease patients using time frequency, cross correlation, Granger casuality and bi-spectral analysis. Results of the time-frequency analysis and cross-frequency correlation analysis demonstrated that the power density of those components significantly decreased as the alleviation of tremor and cross-correlation (0.18-0.50) exists during tremor period. Granger causality of the time-variant amplitude showed stronger contribution from tremor to double tremor components, and contributions from both tremor and double tremor components to triple tremor component. Quadratic phase couplings among these three components were detected by the bispectral approaches. The linear and nonlinear relationships existed among the multi-components and certainly confirmed that the dependence cross those frequencies and neurological mechanism of tremor involved complicate neural processes.
Action Potentials
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Electromyography
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Humans
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Parkinson Disease
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physiopathology
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Subthalamic Nucleus
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physiopathology
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Tremor
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physiopathology
4.Effects of acupuncture and moxibustion on histamine and inflammatory factors in the skin tissue at Tianshu (ST25) of rats with Crohn disease
Jingying ZHOU ; Yitian LAI ; Zongbao YANG ; Xuan XU ; Zhiyi LIANG ; Panting DING ; Guoshan ZHANG ; Mi LIU
Journal of Acupuncture and Tuina Science 2023;21(2):101-108
Objective: To explore the potential mechanism of acupuncture and moxibustion in treating Crohn disease (CD) by evaluating the changes in histamine and inflammatory factors in the skin tissue at Tianshu (ST25) of rats.Methods: Fifty-eight Sprague-Dawley rats were randomly divided into a normal group (n=14) and a CD-modeling group (n=44). Rats in the CD-modeling group received enema with 2,4,6-trinitrobenzene sulfonic acid plus ethanol to establish CD models. The enema was repeated once every 7 d for a total of 4 times. After modeling, four modeled rats and four normal rats were randomly selected for model identification. After the CD model was successfully established, the remaining rats in the CD-modeling group were randomly divided into a model group, an acupuncture group, a moxibustion group, and a Western medication group, with ten rats in each group. The rats in the acupuncture and moxibustion groups were treated with acupuncture or moxibustion at Tianshu (ST25) and Shangjuxu (ST37); the rats in the Western medication group were treated with mesalazine enteric-coated tablets by gavage for continuous 7 d. After the intervention, the colon tissue of rats in each group was collected. After gross observation, hematoxylin-eosin (HE) staining was performed to further observe the pathological changes. The expression of histamine in the skin tissue at Tianshu (ST25) was detected by enzyme-linked immunosorbent assay. The expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-18, IL-10, and IL-6 in the skin tissue at Tianshu (ST25) was detected by Western blotting. Results: Compared with the normal group, the colonic wall of rats in the model group showed cobblestone-like changes, local ulcers, and polyps in dark red and thickening and hardening. HE staining showed local loss of mucosal epithelial layer and formation of slit-like ulcers, destruction of mucosal glands, edema, and infiltration of inflammatory cells in lamina propria and submucosa, and occasional formation of sarcoid-like granuloma. The levels of histamine and IL-6 were significantly up-regulated (P<0.01, P<0.05), and the levels of TNF-α, IL-18, and IL-10 were significantly down- regulated (P<0.01 or P<0.05) in the skin tissue at Tianshu (ST25) of rats in the model group. Compared with the model group, the pathomorphological damage of the colon tissue of rats in the acupuncture group, moxibustion group, and Western medication group was significantly improved. The levels of histamine and IL-6 were significantly down- regulated (P<0.01, P<0.05), and the level of IL-10 was significantly up-regulated (P<0.01) in the skin at Tianshu (ST25) of rats in the acupuncture group. The levels of histamine and IL-6 were significantly down-regulated (P<0.01, P<0.05), and the levels of TNF-α, IL-18, and IL-10 were significantly up-regulated (P<0.01 or P<0.05) in the skin tissue at Tianshu (ST25) of rats in the moxibustion group. The level of histamine was significantly down-regulated (P<0.01), and the levels of IL-18 and IL-10 were significantly up-regulated (P<0.05, P<0.01) in the skin tissue of rats in the Western medication group. Compared with the acupuncture group, the level of IL-10 in the skin tissue at Tianshu (ST25) of rats in the moxibustion group was significantly up-regulated (P<0.01). Conclusion: The inflammatory responses in the skin tissue at Tianshu (ST25) may be the external manifestation of CD. Significant differences in the regulation of inflammatory responses in the skin tissue at Tianshu (ST25) between acupuncture and moxibustion exist, which may be caused by the differences in the stimulation characteristics between acupuncture and moxibustion.
5.Dynamic changes of rat cartilage ultrastructure in the early process of papain-induced knee osteoarthritis
Wenxiu DUAN ; Zongbao WANG ; Hao ZHANG ; Zhiwei YANG ; Zhilun HU ; Fangjun XU ; Yalin XU ; Dan LIU ; Yan XIE
Chinese Journal of Tissue Engineering Research 2015;(18):2789-2793
BACKGROUND:Papain-induced rat knee osteoarthritis is a common modeling method, which can obtain a stable osteoarthritis model.
OBJECTIVE:To observe the change of ultrastructure of chondrocytes in the early process of papain-induced rat knee osteoarthritis under transmission electron microscope.
METHODS:A total of 18 Sprague-Dawley rats were randomly divided into three groups. Two rats were considered as a normal control group, without intervention. The mixture of papain and L-cysteine was injected in right knee joint cavity of 16 rats to induce osteoarthritis models (osteoarthritis model group). Physiological saline was injected in the left side (physiological saline control group). At 1, 2, 4 and 6 weeks after injection, samples were col ected. Transmission electron microscope was used to observe the change of cartilage ultrastructure of the medial femoral condyle joint.
RESULTS AND CONCLUSION:For the normal control group and physiological saline control group, their cytoplasm contained abundant rough endoplasmic reticulum and mitochondria. After 1 week of injection,
mitochondria vacuoles and light expanded rough endoplasmic reticulum were visible. Two weeks later, lipid droplets appeared, mitochondria degeneration was distinct, vacuolization was serious and its number was reduced, and rough endoplasmic reticulum expansion was obvious. Four weeks later, lipid droplets became increased, and the number of mitochondria decreased significantly. Most of the rough endoplasmic reticula were highly expanded, and part of the rough endoplasmic reticula were dissolved and fractured. Six weeks later, a number of lipid droplets were visible in cytoplasm, most of the mitochondria disappeared, only a smal number of mitochondria existed, and most of the rough endoplasmic reticula were dissolved and fractured. These results confirmed that cartilage ultrastructure changes gradual y in the early process of papain-induced rat knee osteoarthritis under transmission electron microscope.
6.Microbial lipid production by Rhodosporidium toruloides in a two-stage culture mode.
Jintao LIN ; Hongwei SHEN ; Zehui ZHANG ; Cuimin HU ; Guojie JIN ; Haidong TAN ; Zongbao K ZHAO
Chinese Journal of Biotechnology 2010;26(7):997-1002
To shorten the cultivation time and reduce the consumption of raw materials for microbial lipid production, oleaginous yeast Rhodosporidium toruloides AS 2.1389 was cultivated using a two-stage culture mode, in which the cell propagation and lipid accumulation were separated. The yeast cells recovered from the propagation culture were re-suspended in glucose solution for lipid accumulation, through which lipid content over 55% of the dry cell weight was achieved, the longer the propagation stage was, the higher the lipid content. Analysis of the lipid indicated that the long-chain fatty acids with 16 and 18 carbon atoms were major components, suggesting that the lipid can be an alternative feedstock for biodiesel production.
Basidiomycota
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growth & development
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metabolism
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Biofuels
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Cell Culture Techniques
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methods
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Fermentation
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Industrial Microbiology
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methods
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Lipids
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biosynthesis
7.Time-effect relationship and molecular mechanism of electroacupuncture for repair of gastric mucosal lesion.
Yuan ZHANG ; Caichun LIU ; Linyu LIAN ; Jiacheng SHEN ; Qida HE ; Jiaolong CHEN ; Fuqiang MA ; Zongbao YANG
Chinese Acupuncture & Moxibustion 2018;38(7):747-752
OBJECTIVETo dynamically observe the effects of electroacupuncture (EA) on repair of gastric mucosal lesion in rats with gastric ulcer, and to explore the time-effect relationship and molecular mechanism of EA for gastric ulcer.
METHODSA total of 72 SD rats were randomly assigned to a normal group, a model group, a acupoint group and a sham acupoint group, and each group were further divided into a 1-day subgroup, a 4-day subgroup and a 7-day subgroup, 6 rats in each subgroup. The rat model of gastric ulcer was established by using intragastric administration of ethyl alcohol. The rats in the acupoint group were treated with EA at"Zusanli"(ST 36) and"Liangmen"(ST 21); the rats in the sham acupoint group were treated with EA at points 5 mm next to"Zusanli"(ST 36) and"Liangmen"(ST 21); the EA was given 30 min per treatment, once a day. The rats in the normal group and model group were treated with immobilization for 30 min per day, and no EA was given. PR-PCR method was applied to test the expression of proliferating cell nuclear antigen (PCNA) and substance P (SP); Western blot method was applied to test the expression of neurotensin (NT).
RESULTSAfter 1-day treatment, the ulcer index in the model group was higher than that in the normal group (<0.01), and the expression of PCNA, SP and NT was decreased (<0.01, <0.05); compared with the model group and sham acupoint group, the ulcer index was decreased in the acupoint group (both <0.05), and the expression of PCNA and SP was up-regulated (all <0.05) while that of NT was up-regulated (both <0.01). After 4-day treatment, the ulcer index in the model group was reduced but still higher than that in the normal group (<0.05), and the expression of PCNA, SP and NT was up-regulated and higher than that in the normal group (all <0.01); the ulcer index in the acupoint group was similar to that in the normal group (>0.05), and the expression of PCNA and SP was lower than that in the model group (<0.01, <0.05), and the expression of NT was not significantly different from that in the model group (>0.05). After 7-day treatment, the differences of indexes above were not significant among the four groups (all >0.05).
CONCLUSIONEA at acupoints of stomach meridian has two-way regulation on PCNA and SP and improve the expression of NT in different pathological state of gastric ulcer, which could further improve the repair of gastric ulcer.