1.Extraction and Separation of Carotenoids of Hydrogen-Producing Rhodobacter sp.
Xiao-Rong ZHANG ; Chun-Gui ZHAO ; Su-Ping YANG ;
Microbiology 1992;0(05):-
Carotenoids play an important role in regulating the hydrogen production of hydrogen-producing Rhodobacter sp. The carotenoids of hydrogen-producing Rhodobacter sp. grown in acetate medium were extracted by using acetone-methanol (7∶2,V/V) solvent and were separated by using thin-layer chromatography on silica-gel plate. The qualitative and quantitative of the carotenoids were analyzed by spectrometry. The results showed that the carotenoids were completely extracted three times with acetone-methanol (7∶2,V/V) in two hours. The ultrasonication had little effect on yield of carotenoids. The yield of carotenoids was 2.81mg/g wet cell. There were 4 spots on the silica-gel plate in the order of yellow, red, light red and light yellow. Yellow spot and red spot were the dominant composition of carotenoid in Rhodobacter sp. The spectrometry data showed that the yellow and red component might be the spheroidene and spirilloanthin respectively.
2.Therapeutic efficacy and mechanism of heat-sensitive moxibustion for adjuvant treatment of depression in Parkinson disease
Xiao-Mei WANG ; Chun-Jiao RONG ; Fu-Qing ZHANG
Journal of Acupuncture and Tuina Science 2020;18(1):33-39
Objective: To observe the clinical efficacy of heat-sensitive moxibustion for adjuvant treatment of depression in Parkinson disease and explore its mechanism. Methods: A total of 80 patients with Parkinson disease coupled with depression were randomized into an observation group and a control group, with 40 cases in each group. The control group was treated with levodopa and benserazide hydrochloride tablets and paroxetine tablets, while the observation group was treated with heat-sensitive moxibustion on the basis of the medications in the control group. The treatment course was 2 months. The Hamilton depression scale-17 (HAMD-17), unified Parkinson's disease rating scale (UPDRS) and Parkinson's disease quality of life questionnaire-39 (PDQ-39) were scored before and after the treatment, and the efficacy was evaluated after treatment. Levels of patients' serum dopamine (DA), 5-hydroxytryptamine (5-HT), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected before and after the treatment. Results: After treatment, the total effective rate of the observation group was higher than that of the control group (P<0.05). The HAMD-17 scores in the two groups decreased significantly after treatment (both P<0.05), and the score in the observation group was obviously lower than that in the control group (P<0.05). The component scores and total scores of UPDRS in both groups decreased significantly (all P<0.05), and the scores in the observation group were lower than those in the control group (all P<0.05). The score of PDQ-39 in the observation group decreased significantly (P<0.05), and was lower than that in the control group (P<0.05). After treatment, the serum DA and 5-HT levels in the observation group increased significantly (both P<0.05) and the TNF-α and IL-6 levels decreased significantly (both P<0.05), which were statistically different from those in the control group (all P<0.05). Conclusion: Heat-sensitive moxibustion has certain auxiliary effect in treating depression in Parkinson disease, significantly improving clinical symptoms and the quality of life, which may be related to the up-regulation of DA and 5-HT levels and down-regulation of TNF-α and IL-6 levels.
3.A Simple and Dependable Approach to Establish Hypoxic-Ischemic Encephalopathy Model in Neonatal Rat
xiao-juan, YIN ; rong, JU ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To investigate a simple and dependable approach to establish the hypoxic-ischemic encephalopathy model in neonatal rat. Methods Twenty-one neonatal rats of 7 days old were randomly divided into control group,hypoxic group,and hypxic-ischemic group.Every group was randomly divided into 3 hours,6 hours,1 day,3 days,7 days, 14 days,and 21 days group,according to the time of killing.Left common carotid artery of neonatal rats at age of 7 days in hypoxic-ischemic group were ligated.Then,the rats in hypoxic and hypoxic-ischemic group were put in a state of 8% oxygen for 2.5 hours. Brain tissues of rats in 3 groups were observed with HE staining under light microscope.Results In hypoxic-ischemic group,there was found mild brain damage after hypoxic-ischomic 3 hours,the brain lesion was most severe at 1 day,glial cell proliferation was found at 3 days,much neur were losed at 14,21 days,and colloid scar was formed in cortex,striatum and hippocampi.Conclusion The method that left common carotid ontery of neonatal rats were ligated and then put in 8% oxygen for 2.5 hours is simple, rapid and dependable, which can be applied widely.
4.Characters of Culture of Neural Stem Cells from Hippocampus of Neonatal Rats in Different States in Vitro
xiao-juan, YIN ; rong, JU ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To investigate the characters of culture of neural stem cells(NSCs) from neonatal rats in different states in vitro.Methods Forty-two neonatal rats at age 7 days were divided randomly into 3 groups as control group,hypoxic group and hypoxic-ischemic group,each having 14 rats.Forteen rats of every group divided randomly into 7 small groups,each including 3 h,6 h,1 d,3 d,(7 d),14 d and 21 d,according to the time to put to death,each having 2 rats.After builting rat models of hypoxic-ischemic encephalopathy,NSCs from hippocampus in 3 groups were isolated,then cultured,passed,differentiated and differentiated with single cell clone and immunocytochemistry tecnique.Results NSCs in hippocampus from 3 groups were cultured in form of typical neuraospheres in suspension.The cells could be cloned,passed continuously and induced.There were differences among 3 groups when primary NSCs were cultured at 3 h and 6 h time points.But at 1 d,3 d,7 d,14 d and 21 d time points,clony neuraospheres from primary NSCs in hypoxic group were the most among 3 groups while clony neuraospheres from primary NSCs in hypoxic-ischemic group were the lest.Conclusions NSCs from hippocampus of neonatal rats in different states remain to be cultured,meanwhile,NSCs are decreased with increase of age,elongation of illness course and progress of state of an illness.
6.Expression of STEAP4 Gene during the Period of Human Preadipocyte Differentiation
xiao-hui, CHEN ; ya-ping, ZHAO ; chun-lin, GAO ; chun-mei, ZHANG ; chun, ZHU ; jin-gai, ZHU ; xi-rong, GUO
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa
9.Effect of Bushen Tongdu Capsule on RANK/RANKL/OPG pathway of collagen induced arthritis rats.
Yang-Chun ZHU ; Lin LIN ; Xiao-Li ZHOU ; Rong-Fang LI ; Li-Ping HOU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(12):1487-1491
OBJECTIVETo study the effect of Bushen Tongdu Capsule (BTC) on RANK/RANKL/ OPG pathway of collagen induced arthritis (CIA) rats, thereby laying theoretic evidence for treating rheumatic arthritis (RA) by Chinese medicine.
METHODSRA model was induced by CIA. Totally 42 rats were randomly divided into six groups, i.e., the normal control group, the model group, the low dose BTC (BSL) group, the medium dose BTC (BSM) group, the high dose BTC (BSH) group, and the Tripterygium Glycosides (TG) group, 7 in each group. BTC at the daily dose of 120, 240, and 480 mg/kg was given by gastrogavage to rats in the BSL, BSM, and BSH group respectively from the 13th day of modeling. TG at the daily dose of 24 mg/kg was given by gastrogavage to rats in the TG group. All medication was given once daily, 2 mL each time. Two mL normal saline was administered to rats in the normal control group and the model group. All medication lasted for 18 days. Samples were taken at day 31. The TRAP section of the ankle joint was fixed in 10% formalin for TRAP stain. Serum levels of osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand (RANKL), and macrophage colony-stimulating factor (M-CSF) were detected using ELISA.
RESULTSCompared with the normal control group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree were significantly improved in the model group, serum levels of RANKL and M-CSF were up-regulated, levels of OPG and OPG/RANKL were significantly lowered (all P < 0.01). Compared with the model group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree also significantly decreased in the BSH group and the TG group (all P < 0.01). RANKL and M-CSF were significantly down-regulated in each medicated group, while levels of OPG and OPG/RANKL were significantly up-regulated (all P < 0.01). Compared with the TG group, M-CSF was lower, but levels of OPG and OPG/RANKL were significantly up-regulated in the normal control group (all P < 0.01). RANKL and M-CSF were significantly up-regulated, while levels of OPG and OPG/RANKL were significantly down-regulated in the model group and each BS group (all P < 0.01).
CONCLUSIONBTC could relieve bone damage of CIA rats possibly through regulating and controlling osteoclasts.
Animals ; Arthritis, Experimental ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Macrophage Colony-Stimulating Factor ; metabolism ; Osteoclasts ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Tripterygium
10.Cloning,Sequence Analysis and Expression in E.coli of the EP0 Gene of Pseudorabies Virus Ea Strain
Liu-Rong, FANG ; Huan-chun, CHEN ; Shao-bo, XIAO ; Xiang-Ru, MA ; Ge-fei, WANG
Virologica Sinica 2001;16(2):183-187
The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea,and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.