3.Feasibility of bacterial cellulose membrane as biological scaffold for construction of tissue engineering corneal epithelium
Jingjie, CAO ; Chen, ZHANG ; Shaozhen, ZHAO ; Yizao, WAN ; Da, HU
Chinese Journal of Experimental Ophthalmology 2016;34(2):121-124
Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100%.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.
5.Extracorporeal Shock Wave Promotes Postnatal Human Bone Marrow Stromal Cells Osteogenesis In vivo
Jun HU ; Da XING ; Aibin ZHANG ; Jiangnan ZHOU
Progress in Biochemistry and Biophysics 2006;0(05):-
It is known that extracorporeal shock wave (SW) may promote healing of fracture. A previous study reported that SW promoted human bone marrow stromal cells (hMSCs) towards osteoblasts in vitro. To study the osteogenesis ability of hMSCs treated by shock wave in porous hydroxyapatite (HA) in vivo, primary hMSCs of SW group and control group were cultured in the porous HA for 2 weeks and then implanted into subcutaneous sites of nude mouse. These implants were harvested and prepared for the biochemical analysis of alkaline phosphatase activity by AKP kit, histological analysis of decalcified and undecalcified sections and morphology by scan electric microscope (SEM), as well as osteocalcin mRNA expression by RT- PCR 4 weeks and 8 weeks after implantation. It showed that cells of SW and control group almost covered the rough surface of HA before implantation and the extracelluar matrix of SW group was abundant by SEM photomicrograph . The histological analysis and SEM photomicrograph showed active bone formation 4 weeks and 8 weeks after implantation, as well as tetracycline labeling under fluoroscopy analysis in SW group. Alkaline phosphatase in supernatants of the implants detected 4 weeks and 8 weeks after implantation in SW group was higher than in control group (P
6.Isolation and differential expression of a novel MAP kinase gene DoMPK4 in Dendrobium officinale.
Gang ZHANG ; Yi-Min LI ; Ben-Xiang HU ; Da-Wei ZHANG ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2014;49(7):1076-1083
Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence
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DNA, Complementary
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genetics
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DNA, Plant
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genetics
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Dendrobium
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enzymology
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genetics
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Gene Expression Regulation, Plant
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Mitogen-Activated Protein Kinases
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genetics
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metabolism
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Phylogeny
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Plant Leaves
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metabolism
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Plant Proteins
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genetics
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metabolism
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Plant Roots
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metabolism
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Plant Stems
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metabolism
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Seeds
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metabolism
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Sequence Alignment
7.Study of the follow-up management strategies after installation of improved stoves to prevent coal-burning type endemic fluorosis in Guizhou province
Bo-you, ZHANG ; Da-sheng, LI ; Yin, LIANG ; Xiao-qiang, HU ; Rui-zhi, ZHANG
Chinese Journal of Endemiology 2011;30(6):697-700
Objective To study the follow-up management strategies after improving stoves for controlling coal-burning type endemic fluorosis in Guizhou and to provide a scientific basis for exploring the follow-up management measures.Methods In 2006 - 2009,three counties of Puding,Bijie and Liuzhi with improved stoves in 2005 and implemented follow-up management measures for subsequent three years were chosen,3 towns were chosen randomly in each chosen county,2 villages were selected randomly in each chosen town,10 households were investigated randomly in each chosen village,and 20 students were investigated of the knowledge of fluorosis control.The investigation included also the usage of the stoves,dehydration methods of grain and related conditions.Results ①The head of the family and student awareness of fluorosis control were 82.6%(743/900)and 91.7% (1650/1800),respectively,and 35.0% ( 385/1100 ) and 61.0% (6605/10 835 ),respectively,before the test,and the difference was statistically significant( x2 =33.04,1189.12,all P < 0.01 ).②The rates of chimney out of the house were 88.5%(69/78) and 100.0%(102/102),respectively,compared with those before the trial[74.4%(2125/2856) and 2.1%(104/4984),respectively],the difference was statistically significant(x2 =720.56,4295.38,all P < 0.01).The iron stove and the table stove's airtight utilization rates were 85.9%(67/78) and 100.0%(102/102),respectively.③The corn and the hot pepper's correct drying rates were 100.0%(180/180).Compared with those[27.2%(49/180) and 32.2%(58/180),respectively]before the trial,the difference was statistically significant(x2 =26.68,37.38,all P < 0.01 ).The corn and the hot pepper's washing rates before eating were 95.0%(57/60) and 98.3%(177/180),respectively.Compared with those[85.0%(153/180) and 77.8%(140/180)]before the trial,the difference was statistically significant(x2 =135.00,490.82,all P < 0.01 ).Conclusions The target population's awareness and knowledge of fluorosis prevention and related behavior are significantly elevated and enhanced after implementation of the three years post-management,which has reached the desired goal.The strategies of the follow-up management suits Guizhou province and widespread application is suggested.
8.Usefulness of Ankle Brachial Index to Predict the Extent of Coronary Stenosis
Shi-wei YANG ; Da-yi HU ; Xian WANG ; Shouyan ZHANG ; Jian ZHANG ; Shi GUO
Chinese Journal of Rehabilitation Theory and Practice 2006;12(6):461-463
ObjectiveTo examine the relationship between ankle brachial index (ABI) and the extent of coronary stenosis and evaluate the usefulness of ABI to predict the extent of coronary stenosis in old patients.Methods118 patients with coronary angiography were examined by ABI and hemostatic factors evaluation in addition to history collection.ResultsABI was inversely and significantly associated with Gensini score. ABI reduced significantly (P<0.001) in the patients with 3-vessel or left main coronary artery disease (CAD). But there were no significant differences in ABI among the patients with no CAD, 1-vessel or 2-vessel CAD. The corresponding area under the ROC curve was 0.75±0.045, with 95% CI=0.67~0.84 (P<0.001) in ABI in 3-vessel or left main CAD. When ABI≤0.9, it had a relatively high specificity (89.1%) and sensitivity (55.6%) for predicting the presence of 3-vessel disease or left main CAD.ConclusionIn the old patients, ABI is inversely and significantly associated with the extent of coronary stenosis, and ABI≤0.9 has a relatively high specificity and sensitivity for predicting the presence of 3-vessel or left main CAD.
9.Exploring the relationship between red blood cell levels and emotional regulation through the miR191-Riok3-Mxi1 pathway
Da Zhang ; Xiaoqing Zhang ; Li Yang ; Yan Zhao ; Xingang Hu
Journal of Traditional Chinese Medical Sciences 2024;11(1):101-110
Objective:
To assess emotional fluctuations, physical and mental health status, and indicators closely related to red blood cells, such as RIO kinase 3 (Riok3), MAX interactor 1 (Mxi1), and microRNA 191 (miR191), in participants with different levels of red blood cells.
Methods:
Participants who underwent physical examinations at Dongfang Hospital between April and October 2019 were divided into healthy, blood deficiency, and anemia groups (30 individuals in the healthy and blood deficiency group respectively, and 13 in the anemia group). The physical and mental conditions of the participants were evaluated through questionnaires, and emotional fluctuations were assessed through an emotion-inducing experiment, in which participants watched video segments designed to induce specific emotions. Relative expression levels of miR191, Riok3, and Mxi1 from venous blood samples were also determined.
Results:
The main psychological factors identified in the anemia and blood deficiency groups were obsessive-compulsive symptoms, depression, anxiety, and other negative emotions. Relative gene expression levels indicated that miR191 was upregulated and Riok3 and Mxi1 were downregulated in both the blood deficiency and anemia groups. Regarding the emotional score of disgust on video stage, the main effect was significant (F = 335.58, P < .001), which showed that watching the three videos caused participants to have a dominant emotion, and there is a difference on group (F = 5.35, P = .01), with higher disgust scores in the anemia and blood deficiency groups. The symptoms of blood deficiency and anemia, such as weakness in limbs were significantly negatively correlated with Riok3 and Mxi1 expression (r = −0.38 and −0.31 respectively), but was significantly positively correlated with miR191 expression (r = 0.29).
Conclusion
We determined that a close relationship exists between red blood cell levels and emotional status. Our findings suggest that individuals with anemia and blood deficiency are more likely to experience psychological problems and negative emotions, particularly disgust. We also demonstrate that emotional regulation is related to mir191-Riok3-Mxi1 pathway activity, identifying these pathway components are potential targets for genetic therapies in combination with psychological therapy.
10.Protective effect of melatonin on oxidative stress inducing hair follicle injury in scald rat.
Jun ZHANG ; Da-Hai HU ; Gang CHEN ; Xiao-Zhi BAI ; Chao-Wu TANG
Chinese Journal of Burns 2009;25(2):129-132
OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.
METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.
RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).
CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.
Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley