1.Effect of compound decotion on activities of Na+-K+-ATPase and Ca2+-ATPase and regulation of Ca2+ after traumatic brain injury
Jingkun MIAO ; Xiaomei WU ; Qixiong CHEN ; Xiaoping ZHANG
Chinese Journal of Trauma 2008;24(12):1002-1006
Objective To dynamically observe the effect of compound decotion on changes of Na+-K+-ATPase,Ca2+-ATPase activity,intracellular free Ca2+ contents and CaM expression jn bomogenate and mitochondria of rat brain tissues after traumatic brain injury(TBI)and investigate the molecular mechanism of neuroprotective effect of compound decotion.Methods Rat TBI models were made and divided into sham operation group,TBI group and compound decotion treatment group(treated with comof normal saline,twice per day for seven days.Rats were sacrificed at 24 h,72 h and 1 week after injury to dynamically observe activities of Na+-K+-ATPase and Ca2+-ATPase in bomogenate and mitochondria of rat brain tissues,concentration of free Ca2+ in neurocytes and expression change of CaM in brain tissues.Results The activities of Na+-K+-ATP ageand Ca2+-ATPase in homogenates and mitochondria of brain tissues markedly decreased at different time point and increased gradually after 72 hours in TBI group but wag still lower than that of sham operation group at one week after injury.However,compound decotion could significantly enhance the activities of Na+-K+-ATPageand Ca2+-ATPage(P<0.05).In TBI group,concentration of free Ca2+ in neurocytes and CaM expression in brain tissues were elevated at different degrees at different time point and reached peak at 24 hours after injury but still lower than that of sham operation group at 72 hours.While concentration of free Ca2+ in neurocytes and CaM expression in brain tissues were significantly lower than those of TBI group at different time point(P<0.05).Conclusions The neuroprotective effect of compound decotion may be related to its role in increasing activities of Na+-K+-ATPase and Ca2+-ATPase to facilitate cellular metabolism and decreasing concentration of free Ca2+ in neurocytes and CaM expression in brain tissues to mitigate secondary brain injury induced by Ca2+ over load.
2.Experimental study on the antiepileptic properties of alpha-asarone in different epilepsy models
Jingkun MIAO ; Xiaomei WU ; Qixiong CHEN ; Xiaoping ZHANG
Chinese Pharmacological Bulletin 2003;0(12):-
Aim To investigate the antiepileptic activity of alpha-asarone in three epilepsy models.Methods The MES mice,MST mice and Lithium-pilocarpine rats were divided randomly and respectively into groups each containing 20 animals(?-asarone groups,AED groups and normal control group).Different doses of alpha-asarone were administered to mice/rats in advance in alpha-asarone treated groups,one group received only saline,while the other groups received antiepileptic drug as a reference standard,2 times per day for 28 days.The seizure severity score,seizure latency and total number of animals with seizures were noted to observe whether alpha-asarone had anticonvulsant effect or not in three epilepsy models.Results Alpha-asarone possessed excellent anticonvulsant effect in MES and MST and lithium-pilocarpine models. It significantly decreased the seizure incidence 40%~100% in the MES models and 50%~90% in MST models,and 40%~80% in the Lithium-pilocarpine model. It significantly prolonged the seizure latency 70~180 s in MST mice and 4~15 min in Lithium-pilocarpine rats;It significantly reduced the seizure severity scores 1.96 in Lithium-pilocarpine rats.Conclusions Alpha-asarone had a positive antiepileptic activity.
3.Neonatal intrahepatic cholestasis caused by citrin deficiency:updated understanding
Jingying LIU ; Jingkun MIAO ; Yuanyuan HUA ; Huijuan LIU ; Qian MA ; Chun LI ; Jialin YU ; Qixiong CHEN
Journal of Clinical Pediatrics 2016;34(12):949-953,955
Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disease caused by SLC25A13 gene mutations, and is characterized by delayed jaundice clearance, liver dysfunction, and elevated aminoacidemia. The confirmed diagnosis depends on gene analysis. Citrin deficiency is one of the important causes of neonatal intrahepatic cholestasis in China. Recently more and more researches about NICCD were reported. The paper summarized the epidemiology, pathogenesis, clinical characteristics, and progresses in diagnosis and treatment of NICCD.
4.Human bone morphogenetic protein 3(hBMP3) inhibits growth of human osteosarcoma cells MG63 and U2OS
Limei WU ; Huanling HE ; Xingxing LI ; Yinghua CHEN ; Jia WEI ; Guowei ZUO ; Jingkun MIAO ; Yan WANG ; Lan ZHOU
Basic & Clinical Medicine 2006;0(07):-
Objective The biological effects of BMP3 on osteosarcoma were investigated by treating human osteosarcoma cell lines MG63,and U2OS with human BMP3(hBMP3).Methods Osteosarcoma cells in experimental groups were respectively treated with AdBMP-3 and rhBMP3-CM,control groups with AdGFP and rGFP-CM,the blank group with neither.Their ability of proliferation,apoptosis,transmigration and differentiation were respectively detected by trypan blue exclusion test,terminal deoynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) and acridine orange-ethidium bromide fluorescent stain(AO/EB),transwell-room and alkaline phosphatase(ALP) activity reagent kit.Results(1) All the indexes detected were not significantly different between two control groups.(2) Compared with control groups,the cell survival rate showed a significant decrease in experimental groups.(3) The apoptosis indexes increased.(4)The number of trans-membrane cell decreased.(5)The activity of alkaline phosphatase increased after treatment with AdBMP3 and rhBMP-3 for 3 days in MG63,5 days in U2OS.Conclusion hBMP3 inhibit osteosarcoma cells growth and promote bone formation.
5.Effect of human bone morphogenetic protein 2, 3, 6, and 12 on osteosarcoma cell line UMR106.
Xingxing LI ; Huanling HE ; Limei WU ; Wen MA ; Yinghua CHEN ; Jia WEI ; Jingkun MIAO ; Lan ZHOU
Journal of Central South University(Medical Sciences) 2010;35(5):464-469
OBJECTIVE:
To investigate the effect of human bone morphogenetic protein (hBMPs) 2/3/6 and 12 on osteosarcoma cell UMR106.
METHODS:
Adenovirus-BMP2/3/6 and 12 (AdBMP2/3/6 and12) were used to treat the cell line. Their proliferation, apoptosis, and transmigration were detected by Trypan blue exclusion test, TdT-mediated biotinylated-dUTP nick end labeling (TUNEL), acridine orange-ethidium bromide (AO/EB) double fluorescent dye staining, and transwell-room test, respectively. The alkaline phosphatase (ALP) activity was detected to reflect the differentiation of tumors.
RESULTS:
Compared with the control groups, the cell survival rate of the experimental groups treated with AdBMP2/3/6 and 12 showed a significant time-dependent decrease (P<0.01). The apoptosis indexes were increased significantly (P<0.01) and the results from TUNEL and AO/EB method were consistent. The cell numbers of transmembrane significantly decreased at 24,48, and 72 h (P<0.01). AdBMP2/3/6 and 12 treatment enhanced the activity of ALP activity from day 3 and this effect might still be observed up to day 9 of the treatment (P<0.01).
CONCLUSION
hBMPs2/3/6 and 12 can inhibit the proliferation and transmigration, and induce their apoptosis and differentiation in osteosarcoma cell line UMR106.
Adenoviridae
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genetics
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metabolism
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Apoptosis
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drug effects
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Bone Morphogenetic Protein 2
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pharmacology
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Bone Morphogenetic Protein 3
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pharmacology
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Bone Morphogenetic Protein 6
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pharmacology
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Bone Morphogenetic Proteins
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pharmacology
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Bone Neoplasms
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pathology
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Cell Line, Tumor
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Cell Transformation, Neoplastic
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drug effects
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Growth Differentiation Factors
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pharmacology
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Humans
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Osteosarcoma
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pathology
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Recombinant Proteins
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pharmacology
6.Covalent organic nanospheres as a fiber coating for solid-phase microextraction of genotoxic impurities followed by analysis using GC-MS
Zhao YANFANG ; Li JINGKUN ; Xie HANYI ; Li HUIJUAN ; Chen XIANGFENG
Journal of Pharmaceutical Analysis 2022;12(4):583-589
Covalent organic nanospheres(CONs)were explored as a fiber coating for solid-phase microextraction of genotoxic impurities(GTIs)from active ingredients(AIs).CONs were synthesized by an easy solution-phase procedure at 25℃.The obtained nanospheres exhibited a high specific surface area,good ther-mostability,high acid and alkali resistance,and favorable crystallinity and porosity.Two types of GTIs,alkyl halides(1-iodooctane,1-chlorobenzene,1-bromododecane,1,2-dichlorobenzene,1-bromooctane,1-chlorohexane,and 1,8-dibromooctane)and sulfonate esters(methyl p-toluenesulfonate and ethyl p-toluenesulfonate),were chosen as target molecules for assessing the performance of the coating.The prepared coating achieved high enhancement factors(5097-9799)for the selected GTIs.The strong affinity between CONs and GTIs was tentatively attributed to T-T and hydrophobicity interactions,large surface area of the CONs,and size-matching of the materials.Combined with gas chromatography-mass spectrometry(GC-MS),the established analytical method detected the GTIs in capecitabine and imatinib mesylate samples over a wide linear range(0.2-200 ng/g)with a low detection limit(0.04-2.0 ng/g),satisfactory recovery(80.03%-109.5%),and high repeatability(6.20%-14.8%)and reproducibility(6.20%-14.1%).Therefore,the CON-coated fibers are promising alternatives for the sensitive detection of GTIs in AI samples.
7.Alternative Biological Material for Tissue Engineering of the Vagina: Porcine-Derived Acellular Vaginal Matrix
Yanpeng TIAN ; Yibin LIU ; Yanlai XIAO ; Zhongkang LI ; Mingle ZHANG ; Liang CHEN ; Zhen LI ; Wangchao ZHANG ; Zhiqiang ZHANG ; Desheng KONG ; Li MENG ; Yanfang DU ; Jingkun ZHANG ; Jingui GAO ; Xianghua HUANG
Tissue Engineering and Regenerative Medicine 2024;21(2):277-290
BACKGROUND:
Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is a severe congenital disorder characterized by vaginal hypoplasia caused by dysplasia of the Müllerian duct. Patients with MRKH syndrome often require nonsurgical or surgical treatment to achieve satisfactory vaginal length and sexual outcomes. The extracellular matrix has been successfully used for vaginal reconstruction.
METHODS:
In this study, we developed a new biological material derived from porcine vagina (acellular vaginal matrix, AVM) to reconstruct the vagina in Bama miniature pigs. The histological characteristics and efficacy of acellularization of AVM were evaluated, and AVM was subsequently transplanted into Bama miniature pigs to reconstruct the vaginas.
RESULTS:
Macroscopic analysis showed that the neovaginas functioned well in all Bama miniature pigs with AVM implants. Histological analysis and electrophysiological evidence indicated that morphological and functional recovery was restored in normal vaginal tissues. Scanning electron microscopy showed that the neovaginas had mucosal folds characteristics of normal vagina. No significant differences were observed in the expression of CK14, HSP47, and a-actin between the neovaginas and normal vaginal tissues. However, the expression of estrogen receptor (ER) was significantly lower in the neovaginas than in normal vaginal tissues. In addition, AVM promoted the expression of b-catenin, c-Myc, and cyclin D1. These results suggest that AVM might promotes vaginal regeneration by activating the b-catenin/cMyc/cyclin D1 pathway.
CONCLUSION
This study reveals that porcine-derived AVM has potential application for vaginal regeneration.
8.Clinical characteristics of extremely premature infants at low risk for early-onset sepsis
Yuanyuan HUA ; Jingkun MIAO ; Xian ZOU ; Xin LAI ; Na CHEN ; Linchao YU ; Qixiong CHEN
Journal of Clinical Pediatrics 2019;37(1):1-5
Objective To explore and analyze the clinical characteristics of extremely premature infants at low risk for early-onset sepsis (EOS) , so as to avoid overuse of antibiotics. Method The clinical data of extremely premature infants hospitalized from January 1, 2010 to December 31, 2017 were collected. Extremely premature infants born from mothers without premature rupture of membranes and without maternal clinical manifestations of chorioamnionitis during pregnancy were classified assigned into the low-risk group, and those who did not meet the low-risk conditions were regarded assigned intoas the control group. EOS was diagnosed according to the results of blood culture within 72 hours after birth. The clinical characteristics, treatment and outcome of extremely premature infants between the two groups were retrospectively analyzed. Results A total of 245 extremely preterm infants were enrolled, including 153 (62.4%) in low-risk group. Compared with the control group, mothers in low-risk group had higher rates of gestational diabetes and hypertension, higher rates of antenatal hormone use and lower rates of antenatal antibiotics use; furthermore, neonates in low-risk group had lower rates of Apgar score < 5, higher rates of pulmonary surfactant use, respiratory support and mechanical ventilation, and lower risk of death and incidence of early-onset sepsis. The differences were statistically significant (P<0.05) . Among In extremely premature infants whose having survival time> 24 hours, compared with control group, infants in low-risk group had higher incidences of respiratory distress syndrome, patent ductus arteriosus, intracranial hemorrhage and bronchopulmonary dysplasia, and lower incidence of pulmonary hemorrhage than control group, and the differences were statistically significant (all P<0.05) . In low-risk group, the risks of death, respiratory distress syndrome, pulmonary hemorrhage and bronchopulmonary hemorrhage in long-term antibiotic group were higher than the short-term antibiotic group. Conclusion Early identification of extremely preterm infants at low risk of early-onset sepsis in extremely preterm infants is of clinical significance in reducing early empirical use of antibiotics therapy.
9.Preparation and Quality Evaluation of Zingiber officinale Oil Microcapsules
Xianzhe FAN ; Fulin HE ; Xiaowen LIU ; Jingkun LYU ; Xiaoming CHEN
China Pharmacy 2019;30(21):2920-2925
OBJECTIVE: To prepare Zingiber officinale oil microcapsules and to evaluate its quality. METHODS: Z. officinale oil microcapsules were prepared by spray drying method with sodium starch octenyl succinate as capsule material. The preparation technology was optimized by orthogonal test with mixing temperature of capsule material and capsule core, mass ratio of capsule material and capsule core, stirring speed as factors, using encapsulation efficiency as index. The drug loading, encapsulation efficiency, appearance, particle size distribution and stability of light, heat and humidity (using iodine value and peroxide value as indexes) were evaluated. RESULTS: The optimal preparation technology of Z. officinale oil microcapsules was that the mixing temperature of capsule material and core was 60 ℃; mass ratio of capsule material and capsule core was 10 ∶ 1; stirring speed was 12 000 r/min. Average drug-loading amount and encapsulation efficiency of Z. officinale oil microcapsules prepared by optimal technology were 17.97% and 73.57% (n=3). The morphology of Z. officinale oil microcapsules was round, smooth, non-sticky and uniform in size distribution. The average diameter of microcapsules was (6.30±0.27) μm. Under light, heat and humidity conditions, the iodine value and peroxide value of Z. officinale oil microcapsules changed slightly. CONCLUSIONS: The optimal preparation technology of Z. officinale oil microcapsules is simple and reproducible. The prepared microcapsules have good encapsulation efficiency, high drug loading amount and good stability.
10.BMSCs promote M2 macrophage polarization to attenuate acute radiation-induced lung injury
Xinhui ZHANG ; Shiying NIU ; Shutong YAO ; Xiaoyue ZHANG ; Xuetao CAO ; Xue GAO ; Guoli ZHAO ; Jingkun CHEN ; Yueying ZHANG
Chinese Journal of Radiological Health 2024;33(1):21-27
Objective To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) for radiation-induced lung injury (RILI) and the underlying mechanism. Methods Forty-five healthy adult male C57BL/6 mice were randomly divided into control, model, and BMSCs groups. The model and BMSCs groups received a single irradiation dose of 20 Gy to the chest, while the control group did not receive X-ray irradiation. For the BMSCs group, an injection of 1 × 106 BMSCs cells was administered via the tail vein within 6 h after irradiation. In the 5th week, the lung tissue was taken to observe pathological changes with HE staining; examine the expression of the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) with immunohistochemical staining; observe the polarization of macrophages with immunofluorescence staining; and measure the expression of the epithelial-mesenchymal transition markers E-cadherin, N-cadherin, and vimentin proteins by Western blot. Results After radiation, the model group developed pulmonary vasodilation and congestion with septal thickening and inflammatory cell infiltration, and these changes were markedly reduced in the BMSCs group. The model group showed significantly down-regulated expression of IL-6 and TNF-α compared with significantly increased levels in the model group (P < 0.01, P < 0.05). Treatment with BMSCs significantly increased the polarization of lung macrophages towards the M2 type, while significantly decreasing the abnormally increased N-cadherin and vimentin levels in RILI mice (P < 0.05, P < 0.01). Conclusion BMSCs have therapeutic effects for RILI mice, which may be through promoting macrophage polarization from M1 to M2.