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Table of Content

    20 December 2019, Volume 15 Issue 6 Previous Issue    Next Issue
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    Original article
    Preliminary Research of Liposome in modRNA Transfection to Different Cell Types
    YAN Bingqian,WANG Huijing,AI Xuefeng,GONG Yiqi,TAN Yao,XU Xu,WANG Wei,FU Wei
    2019, 15 (6):  373-383.  DOI: 10.3969/j.issn.1673-0364.2019.06.001
    Abstract ( 561 )  
    Objective To screen a better liposome agent for modRNA transfection and to explore the feasibility and efficiency of using it to transfect various cell types and express target proteins. Methods After modGFP was transfected into MEF cells with RNAiMax and MessageMax, the transfection efficiency and protein expression effects of the two transfection agents were detected and compared by fluorescence microscope and flow cytometry. The better liposome agent was chosen for modGFP transfection into various cell types, and the transfection and expression efficiency were measured using the same methods. Besides, the co-transfection of modGFP and modmCherry, and the nuclear transfection of modnGFP and modmTBX5 into MEF cells were observed by fluorescence microscope. Results After MEF cells were transfected with modGFP for 24 h, flow analysis showed that the transfection efficiency of MessageMax(83.33%±3.23%) was slightly higher than that of RNAi Max(78.77%±6.12%), but there was no statistical difference. However, both flow analysis and fluorescent images found that MessageMax had a higher efficiency of protein expression within 1 week after transfection, making it a better modRNA transfection reagent. MessageMax could efficiently transfect modGFP into 3T3 cells, Hela cells and MCF-7 cells, achieve co-translocation of modGFP and modmCherry, and achieve nuclear localization of intranuclear factors nGFP and mTBX5 in MEF cells. Conclusion Compared to RNAi Max, modRNA transfection with MessageMax brings better results, which can not only achieve efficient and rapid expression of various target proteins including nuclear proteins in various cell types, but also realize simultaneous transfection and expression of multiple factors.
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    Fish Collagen in Constructing Tissue-Engineered Cartilage
    ZHAO Shaohua,WANG Yigong,XU Yong,DUAN Liang
    2019, 15 (6):  379-383.  DOI: 10.3969/j.issn.1673-0364.2019.06.002
    Abstract ( 464 )  
    Objective To explore the feasibility of fish collagen in constructing tissue-engineered cartilage. Methods Porous fish collagen was fabricated by lyophilization. Auricular chondrocytes derived from rabbit were seeded into the porous fish collagen. After cultured in vitro for 2 or 4 weeks, the cytocompatibility and the capacity of cartilage-specific extracellular matrix formation based on fish collagen were investigated. Thereafter, the cell-scaffold constructs were implanted into the rude mice subcutaneously. Gross observation, histological observation and quantitative evaluation of neocartilage test were carried out after cultured for 6 weeks. Results Fish collagen possessed satisfactory cytocompatibility, and facilitated cartilage-specific extracellular matrix formation both in vitro and in vivo. Notably, HE staining showed typical mature lacuna after cultured in vitro for 4 weeks or cultured in vivo for 6 weeks. After cultured in vivo for 6 weeks, Safranin-O staining showed abundant proteoglycan, biomechanical and biochemistry analyses indicated that neocartilage was comparable than native cartilage. Conclusion Fish collagen is a promising natural derived material as scaffold in constructing tissue engineering cartilage.
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    Porous Scaffold Based on Egg White for Cartilage Tissue Engineering
    JIA Zihao,LI Hao,XU Yong,ZHOU Guangdong,TAN Xiaoyan,ZHANG Wei
    2019, 15 (6):  384-388.  DOI: 10.3969/j.issn.1673-0364.2019.06.003
    Abstract ( 481 )  
    Objective To explore the feasibility of porous scaffold based on egg white for tissue-engineered cartilage regeneration. Methods Egg white was mixed with deionized water in different volume ratios(1:0, 1:1 and 1:2) to fabricate porous scaffolds after freeze-drying. Afterwards, the three kinds of scaffolds were conducted with gross observation, porous size and mechanical property examination. Chondrocytes derived from rabbit ear in the second passage were seeded into the scaffolds with different volume ratios and cultured in vitro. The examination of Living & dead staining and cell proliferation assay were conducted after in vitro cultivation for 1, 4 and 7 days. Moreover, the chondrogenesis was observed by histological examination after in vitro culture for 3 weeks. Results Porous scaffolds could be fabricated by all the three kinds of egg white with different volume ratios. The pore size presented significantly increasing trend with the increase of deionized water content, but the mechanical property presented the reversed trend. The examination of both living & dead staining and cell proliferation assay showed that the chondrocytes could attach and proliferate on all the three kinds of scaffold. When extended the in vitro culture time to 3 weeks, HE staining showed that egg white scaffolds of all three groups had immature neocartilage matrix formation, which confirmed the feasibility of constructing cartilage based on egg white scaffold in vitro.Notably, with the increase of deionized water ratio, the amounts of residual scaffolds decreased but the component of neocartilage matrix increased. Conclusion Porous scaffolds based on egg white are suitable for constructing tissue engineered cartilage.
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    Effect and Mechanism of Adenosine Triphosphate on Differentiation of Bone Marrow Mesenchymal Stem Cells into Chondrocytes
    LEI Jie,HE Chengjian,HE Mengyin
    2019, 15 (6):  389-392.  DOI: 10.3969/j.issn.1673-0364.2019.06.004
    Abstract ( 456 )  
    Objective To investigate the effect of adenosine triphosphate on the differentiation of BMSCs into chondrocytes and its mechanism. Methods The primary cells of BMSCs from 4-5 weeks old SD rats were cultured to the third generation in vitro. PNPP method was used to detect the effects of ATP with different concentrations(150, 300 and 600μmol/L) and BBG(receptor inhibitor of P2X7) of on alkaline phosphatase expression in BMSCs. The mRNA expression of multiple cartilage markers was detected by semi-quantitative reverse transcription polymerase chain reaction and real time PCR. Results After ATP was added, the expression of alkaline phosphatase in BMSCs was significantly increased(P<0.05).The highest expression of alkaline phosphatase was observed when ATP concentration was 150 and 300 μmol/L(P<0.05).The expression levels of indicators such as SOX6, SOX9 and COL2A1 were increased(P<0.05). After the addition of P2X7 receptor inhibitor, the promotion effect of ATP on alkaline phosphatase decreased(P <0.05), and the expressions of chondrogenic indicators such as SOX6, SOX9 and COL2A1 were also significantly decreased(P <0.05). Conclusion A certain concentration of ATP can promote the differentiation of bone marrow mesenchymal cells into chondrocytes, which may play a role through the mediation of P2X7 receptor.
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    Experimental Research of Tissue Engineering Cartilage Based on Electrospun Human Umbilical Cord Wharton’s Jelly
    LI Hao,XU Yong,XIA Huitang,ZHOU Guangdong,JIANG Gening,LI Chengde
    2019, 15 (6):  393-397.  DOI: 10.3969/j.issn.1673-0364.2019.06.005
    Abstract ( 585 )  
    Objective To explore the preparation of nanofiber membrane by human umbilical cord Wharton’s jelly and to explore its feasibility as a scaffold for cartilage tissue engineering. Methods The human umbilical cord Wharton’s jelly was decellularized and mixed with polycaprolactone(PCL) to prepare an electrospun nanofiber membrane. The cytocompatibility of the nanofiber membrane was verified by implantation with chondrocytes derived from rabbit ear in vitro. Thereafter, the chondrocytes-nanofiber membrane constructs were implanted in nude mice for 6 weeks to verify the feasibility of cartilage formation in vivo by histological examination. Results After decellularization of Wharton’s jelly, the cellular components were removed and most of the collagen was retained. The porous nanofiber membrane was successfully prepared by mixed Wharton’s jelly with PCL. Live and dead staining and CCK-8 assay confirmed that the nanofiber membrane possessed perfect cytocompatibility. In vivo histological observation confirmed that the nanofiber membrane-chondrocytes construct could regenerate mature cartilage tissue. Conclusion The human umbilical cord Wharton’s jelly can be fabricated into a suitable scaffold by electrospinning technology for cartilage tissue engineering.
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    Preparation of Single-Cell Suspensions from Human Keloid Tissue by Modified Combined Enzyme Digestion
    HUANG Xin,XU Xiangwen,GAO Yashan,LI Haizhou,GU Suchen,KUANG Yimin,ZAN Tao
    2019, 15 (6):  398-402.  DOI: 10.3969/j.issn.1673-0364.2019.06.006
    Abstract ( 522 )  
    Objective To investigate the effect of modified method of combined enzyme digestion for keloid dermis digestion and to explore the optimal digestion time. Methods Eight keloid tissue samples were collected. After the removal of the epidermis, dermis was homogenized with the scissor. The homogenates of the dermis were digested with group A or B protocol simultaneously. The group A formula was composed of traditional combined enzyme(type Ⅰ collagenase + type Ⅱcollagenase + type Ⅳ collagenase + hyaluronidase), while the group B formula was supplemented with DNase Ⅰ. Single-cell fraction and single-cell numbers were determined under the microscope. Cell viability was detected using Trypan blue staining and flow cytometry. The digestive efficiency between group A and B was compared. Meanwhile, the optimal digestion time for group B was investigated. Results After 2 hours and 4 hours digestion, the fraction(P0.05). Compared with 6 hours digestion, there was no significant increase in the single-cell fraction and single-cell number when the digestion time was extended to 8 hours or 10 hours(P>0.05), whereas the cell viability was decreased significantly(P<0.05). Conclusion The supplement of DNase Ⅰ to the traditional combined enzyme digestion can increase the digestive efficiency and thus, facilitate the preparation of singlecell suspension for keloid tissue. Tissue digestion for 6 hours can achieve satisfactory digestive results while maintaining the cell viability.
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    Effect of Hyaluronidase in Treating Hyaluronic Acid Induced Artery Embolism of Auricular Skin flap in Rabbits
    ZHUANG Yan,LIU Chunjun,YANG Mingyong
    2019, 15 (6):  403-421.  DOI: 10.3969/j.issn.1673-0364.2019.06.007
    Abstract ( 536 )  
    Objective To explore the effect of intravascular hyaluronidase(HAase) in treating rabbit auricular skin flap ischemia caused by HA induced artery embolism, and to compare the different effect of intravenous thrombolysis and selective intra-arterial thrombolysis with HAase. Methods The auricular skin flap model of 20 μL HA artery emblism were created in twenty-five rabbits and the rabbits were divided into five groups(n=5). Control group: no treatment; IVT-NaCl group: intravenous thrombolysis with saline; IVT-HAase group: intravenous thrombolysis with HAase; IAT-NaCl group:intra-arterial thrombolysis with saline; IAT-HAase group: intra-arterial thrombolysis with HAase. Fluorescein examination was performed 4 hours, 3 days and 7 days after operation. The difference of the area percentage of flap perfusion in the five groups was compared 7 days after operation. Results The mean area percentage of flap perfusion of IAT-HAase group was significantly higher than that in the other four groups(P 0.05). Conclusion The selective intra-arterial thrombolysis with HAase can effectively improve the ischemia of the flap caused by HA induced artery embolism, increase the tissue perfusion, and reduce the necrotic area of the flap. However, the effect of intravenous use of HAase as thrombolytic agent needs further investigation.
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    Clinical Efficiency and Mechanism Research of Far Infrared Radiation in the Treatment of Chronic Lymphedema of Extremities
    XIAO Wentian,LI Ke,ZHANG Zheng,XI Wenjing,LIU Ningfei,ZHANG Yixin
    2019, 15 (6):  408-412.  DOI: 10.3969/j.issn.1673-0364.2019.06.008
    Abstract ( 687 )  
    Objective To investigate the therapeutic effect of far infrared radiation in the treatment of chronic lymphedema of extremities and to explore the mechanism. Methods Thirty-two patients with chronic lymphedema of extremities in stage Ⅱ and Ⅲ were treated with far infrared radiation. Before and after the treatment, clinical effects related to extremities swelling were evaluated, including the amount of fluid, the measurement of limb circumference, the thickness of dermis and subcutaneous tissue, the quality of life and the adverse reactions. Eleven patients(34%) went through laboratory index evaluation including cytokines and macromolecules associated with the swelling(IL-6, leptin, hyaluronan and total protein) in serum and tissue fluid of the lymphedema extremities. Results After treatment, the clinical evaluation showed the decrease of extracellular fluid(0.763±0.259) Kg(P<0.001), the decrease of fluid in the extremities(1.204±0.737) Kg(P <0.001), the decrease of circumference of extremities(2.0 ±1.3) cm(P <0.001), the decrease of thickness of skin and subcutaneous tissue(0.07±0.10) cm(P<0.001);(0.32±0.35) cm(P<0.001), and the improvement of quality of life(1.375±0.942)(P<0.001). And the laboratory index evaluation found that the concentration of IL-6 in serum was increased(13.947±4.436) pg/mL(P <0.05), the level of leptin in tissue fluid was decreased(11456.210 ±12656.400) pg/mL(P <0.01), and the ExtraM-Hyaluronan and ExtraM-total protein in affected extremity were both decreased(0.267±0.171) g(P<0.01);(0.137±0.083) Kg(P <0.01). Conclusion Far infrared radiation treatment can effectively reduce the water, lipid, protein and hyaluronan deposited in the swollen tissues and improve limb swelling symptoms by increasing the concentration of IL-6 in the body and reducing the level of leptin in the limbs.
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    Clinical Study of the Personalized Nasal Framework Fabrication Aided with Digital Technology
    CHEN Lianjie,LI Kongying,WANG Jue,LI Binhang,ZHOU Xu
    2019, 15 (6):  413-415.  DOI: 10.3969/j.issn.1673-0364.2019.06.009
    Abstract ( 514 )  
    Objective To explore the efficacy of designing and fabricating personalized nasal framework with digital technology. Methods From November 2018 to August 2019, the three-dimensional nasal morphology of 14 patients was reconstructed by digital simulation technology, the preoperative and postoperative predicted three-dimensional nasal morphology was measured and compared, the volume of prosthesis needed for surgery was estimated, and the nasal prosthesis was designed. The nasal framework model was fabricated by three-dimensional printing technology to guide the fabrication of silicone framework. The changes of three-dimensional nasal morphology were measured by digital technique. Results The average follow-up period was 7 months. 12 patients and their parents were satisfied with the nasal shape, 2 cases thought that the shape of the nasal tip was general, and there was no obvious abnormal nasal shape and function. There was statistical difference in the distance between the nasion and nasal tip before and after operation(P =0.04). There was no significant difference in the distance between the nasion and the columella point and the nasolabial angle before and after operation(P=0.74; P=0.56). Conclusion Digital technology can realize the precise design and personalized preparation of nasal prosthesis, provide accurate guidance for the fabrication of nasal prosthesis scaffold, and provide objective evaluation for the three-dimensional structure of nose after plastic surgery.
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    Effectiveness of Three Surgical Methods in the Treatment of Moderate and Severe Blepharoptosis
    ZHANG Zhiyang,ZHOU Lei,LI Wei
    2019, 15 (6):  416-442.  DOI: 10.3969/j.issn.1673-0364.2019.06.010
    Abstract ( 471 )  
    Objective To explore the effect of three surgical methods for the treatment of moderate and severe ble-pharoptosis. Methods From May 2015 to April 2018, ninety patients(120 eyes) with congenital moderate and severe ble-pharoptosis were selected. According to the principle of random grouping, the patients were randomly divided into 3 groups:group A, B and C. Patients in group A were treated by frontal muscle fascia suspension, patients in group B were treated by traditional levator shortening, and patients in group C were treated with levator palpebrae muscle shortening and fascial sheath suspension. After 6 months of treatment, the differences of therapeutic effect, blepharoptosis correction rate, cosmetic effect, surgical indexes and complications were observed. Results The total effective rate and the correction rate of group C were both significantly higher than those of group A and group B(P0.05). Conclusion Combined with fascial sheath suspension in the treatment of moderate and severe ptosis, it can effectively improve the height of palpebral fissure with high safety and significant therapeutic and cosmetic effects.
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    Comparison of Mid-Term Results of Femoral Head Prostheses with Different Diameter for Total Hip Arthroplasty
    ZHANG Xifeng,LI Qiang,YANG Zonghua,LI Meng
    2019, 15 (6):  419-421.  DOI: 10.3969/j.issn.1673-0364.2019.06.011
    Abstract ( 615 )  
    Objective To compare the difference of mid-term outcomes of femoral head prostheses with different diameter for total hip arthroplasty. Methods Retrospective analysis of 42 patients who underwent total hip arthroplasty for end-stage hip disease from March 2010 to September 2013 was conducted. Among them, 21 cases adopted 36 mm large-diameter ceramic femoral head prosthesis(large diameter group), and 21 cases adopted 32 mm small-diameter ceramic femoral head prosthesis(small diameter group). All the patients were followed up for more than 5 years, and differences in efficacy were assessed by Harris hip score and X-ray. Results The mean postoperative follow-up time was(69.71±4.03) months.Up to the last follow-up, the Harris hip score of all patients was significantly higher than that before surgery(84.48±9.06 vs 40.21±9.05, P<0.001), and the Harris hip score of the large diameter group was significantly higher than that of the small diameter group(89.38±9.36 vs 79.57±5.51, P<0.001). The final follow-up X-ray showed that the prosthesis position of all patients was good, and no dislocation of prosthesis was observed in both groups. Conclusion The mid-term outcome of the large-diameter femoral head prosthesis is satisfactory, and the postoperative joint mobility was improved better than that of the small-diameter prosthesis.
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    Nursing Plan of Complications after Ear Reconstruction Using Nagata Technique
    YANG Min,XU Feng,WU Ying
    2019, 15 (6):  422-424.  DOI: 10.3969/j.issn.1673-0364.2019.06.012
    Abstract ( 441 )  
    Objective To explore the nursing measures for complications after ear reconstruction using Nagata technique in microtia patients. Methods Microtia patients undergoing ear reconstruction using autogenous rib cartilage with Nagata technique were retrospectively analyzed. In view of postoperative complications such as hematoma, skin flap necrosis, infec-tion and stent exposure, sequential nursing and surgical dressing change were combined for targeted treatment, and the prog-nosis of postoperative complications was observed. Results A total of 468 cases were retrospectively reviewed. Among all the cases, 18 patients had postoperative complications, including 10 cases of hematoma, 2 cases of skin flap necrosis, 5 cases of infection and 1 case of stent exposure. All of the reconstructed ears achieved wound healing(Class III, b) after careful se-quential nursing and dressing change, and reoperation was avoided. The patients were followed up for half a year and the re-constructed ear was in good shape. Conclusion For postoperative complications of total ear reconstruction using Nagata technique, targeted education, dressing change and sequential nursing can significantly reduce the recovery time of postoper-ative complications and avoid reoperation, which is worthy of clinical promotion and application.
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    Review
    Advances of Cryoprotective Agents for Stem Cells
    ZHANG Tianyu,ZHOU Shuangbai,LI Qingfeng
    2019, 15 (6):  425-427.  DOI: 10.3969/j.issn.1673-0364.2019.06.013
    Abstract ( 660 )  
    Stem cell can promote tissue repair and regeneration, and has a promising potential application in clinic.Cryopreservation is a common way to achieve long-term preservation of stem cells. An effective cryoprotective agents(CPA)is vital to prevent cells from cryodamage and keep cell viability during cryopreservation process. In this paper, the recent advances in CPA for stem cells were reviewed.
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    Animal Scar Model and Its Application in Laser Therapy Experiments
    WANG Jingyan,ZHANG Yixin
    2019, 15 (6):  428-430.  DOI: 10.3969/j.issn.1673-0364.2019.06.014
    Abstract ( 582 )  
    Scar is a difficult problem in plastic surgery. Laser treatment of scar has gradually become a hot spot in recent years. Animal models can help researchers elucidate the underlying mechanism of scar and explore more possible therapies for laser treatment of scar. In this paper, the commonly used animal scar models and their applications in laser therapy experiments were reviewed.
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    Application of Skin Flap in the Repair of Nasal Defect
    WANG Jue,LI Kongyin,CHEN Lianjie,ZHOU Xu
    2019, 15 (6):  431-434.  DOI: 10.3969/j.issn.1673-0364.2019.06.015
    Abstract ( 514 )  
    Nasal defects are common deformities of facial tissue defects in clinic, mainly repaired by flaps at present. To achieve successful repair, the selection of different flaps must consider the location, the layers involved, the size of the defect and the condition of the patients. After reviewing the literature published at home and abroad in recent years, the commonly used flaps were summarized into four types in this paper: local flap, adjacent flap, composite tissue flap and free flap. The indications, advantages, disadvantages and the new progress of various flaps in each type were reviewed.
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    The Clinical Measurement and Classification of Plagiocephaly
    Wong Ka Ioi,WEI Min,YU Zheyuan
    2019, 15 (6):  435-438.  DOI: 10.3969/j.issn.1673-0364.2019.06.016
    Abstract ( 533 )  
    The deformity of plagiocephaly is roughly divided into mechanical and congenital plagiocephaly, the previous one is caused by external force, while the latter can be caused by craniosynostosis or other craniofacial deformities. This kind of disease do not belong to any pathological changes whatever the reason of the plagiocephaly is, the appearance of the patient’s outlook is the main characteristic that takes our high considerations, so as the long-term concern to the mental problems, which may trigger a series of psychological diseases. Thus, early treatment is crucial. In order to have a better understanding of the oblique head deformity, it is necessary to classify the plagiocephaly of different degrees with the aim to give the most appropriate and effective treatment with the evidence provided for the clinical diagnosis and clinical use. In this paper, the clinical measurement and classification of plagiocephaly were reviewed.
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    Research Progress in Surgical Treatment of Alveolar Cleft
    HU Yuehan,SONG Qinggao
    2019, 15 (6):  439-442.  DOI: 10.3969/j.issn.1673-0364.2019.06.017
    Abstract ( 616 )  
    Alveolar cleft is a congenital craniofacial deformity caused by the fusion disorder of medial nasal process and maxillary process during embryonic development. Bone transplantation is the main treatment. At present, in the repair treatment of alveolar cleft, a large number of scholars are still exploring: when to carry out operation, the selection of different repair materials, the effect of adding various auxiliary materials on osteogenesis, and how to avoid the exposure and infection of the operation area. In this paper, the new research progress in the treatment of alveolar cleft was reviewed.
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