1.Preparation and characterization of 3D plant-based scaffold based on decellularization method in liver tissue engineering
Jingjing HU ; Songlin HE ; Daxu ZHANG ; Shuo ZHAO ; Xiaonan SHI ; Weilong LI ; Shujun YE ; Jingyi WANG ; Quanyi GUO ; Li YAN
Chinese Journal of Tissue Engineering Research 2024;28(29):4645-4651
BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.
2.Experience of diagnosis and treatment of fibro-adipose vascular anomaly
Chong XIE ; Huaijie WANG ; Zhengtuan GUO ; Weilong LIN ; Jinbang ZHOU ; Weijia YANG
Chinese Journal of Plastic Surgery 2023;39(11):1175-1182
Objective:To explore the diagnosis, staging, and treatment of fibro-adipose vascular anomaly(FAVA).Methods:The data of the patients with FAVA admitted to Xi’an International Medical Center Hospital between October 2019 and February 2023 were retrospectively reviewed. Ultrasound and magnetic resonance imaging (MRI) were routinely performed. X-ray and CT were performed for patients with unequal length of lower limbs, lesions involving joints, and obvious joint deformities. The treatment plan was made according to the stage: stage Ⅰ (pain stage), open or laparoscopi radical resection of the lesion was performed; stage Ⅱ (contracture stage), radical surgery was performed to remove the lesion, sometimes combined with Achilles tendon lengthening or tenolysis, and rehabilitation training was performed 2 weeks after surgery; stage Ⅲ(deformity stage), comprehensive treatment based on surgical resection was adopted, combined with joint capsule release, Achilles tendon lengthening or dissection, tendon transfer and oral sirolimus (each time 0.08 mg/m 2, twice daily ) before and after the operation. For patients with lesions involving multiple anatomical regions, staged surgery was performed, and in principle, only one anatomical region was operated per time. Patients were followed up for pain, joint activity and recurrence. Results:A total of 42 patients were admitted, including 18 males and 24 females. The onset age was (7.3±5.0) years, but the average age of diagnosis was (12.5±6.0) years. The lower limbs were involved in 38 cases, the upper limbs in 4 cases. There were 17 cases of stage Ⅰ, 17 cases of stage Ⅱ and 8 cases of stage Ⅲ. Only 4 cases had no misdiagnosis experience, and the misdiagnosis rate was 90.5%(38/42). Persistent pain, muscle contractures and joint deformities were the main clinical symptoms of the disease. MRI showed heterogeneous high and low signal intensity on T1-weighted images, and the high signal intensity was the same as that of subcutaneous adipose tissue. T2 fat-suppressed sequences showed stronger heterogeneous hyperintensity. The follow-up time was (14.6±10.8) months. Patients who took sirolimus orally before or after surgery experienced significant relief of pain symptoms. Of the 42 patients, the symptoms of 31 patients were completely relieved after the operation, and 11 patients still had residual pain or joint movement disorder or even deformity after the operation. Sixteen of 17 stage Ⅰ patients were cured, the lesion was further expanded and the pain recurred in 1 case after the operation. Of the 17 stage Ⅱ patients, 15 were cured, and 2 had mild limitation of ankle movement after the operation. Eight stage Ⅲ patients had varying degrees of pain or joint movement disorder after surgery, postoperative oral sirolimus significantly relieved symptoms. All 10 patients with stage Ⅰ and Ⅱ who underwent endoscopic resection were cured.Conclusion:FAVA usually occurred in school-age and adolescent children. Pain, muscle contracture and joint deformity were the characteristic clinical features. Combined with MRI features, the diagnosis can be confirmed. FAVA staging system could guide treatment and predict prognosis. For stage Ⅰ and Ⅱ patients, surgery should be performed as soon as possible and the prognosis was good. For stage Ⅲ patients, surgery-based comprehensive treatment could improve symptoms, but it was difficult to cure them radically. Oral sirolimus was also required after surgery.
3.Experience of diagnosis and treatment of fibro-adipose vascular anomaly
Chong XIE ; Huaijie WANG ; Zhengtuan GUO ; Weilong LIN ; Jinbang ZHOU ; Weijia YANG
Chinese Journal of Plastic Surgery 2023;39(11):1175-1182
Objective:To explore the diagnosis, staging, and treatment of fibro-adipose vascular anomaly(FAVA).Methods:The data of the patients with FAVA admitted to Xi’an International Medical Center Hospital between October 2019 and February 2023 were retrospectively reviewed. Ultrasound and magnetic resonance imaging (MRI) were routinely performed. X-ray and CT were performed for patients with unequal length of lower limbs, lesions involving joints, and obvious joint deformities. The treatment plan was made according to the stage: stage Ⅰ (pain stage), open or laparoscopi radical resection of the lesion was performed; stage Ⅱ (contracture stage), radical surgery was performed to remove the lesion, sometimes combined with Achilles tendon lengthening or tenolysis, and rehabilitation training was performed 2 weeks after surgery; stage Ⅲ(deformity stage), comprehensive treatment based on surgical resection was adopted, combined with joint capsule release, Achilles tendon lengthening or dissection, tendon transfer and oral sirolimus (each time 0.08 mg/m 2, twice daily ) before and after the operation. For patients with lesions involving multiple anatomical regions, staged surgery was performed, and in principle, only one anatomical region was operated per time. Patients were followed up for pain, joint activity and recurrence. Results:A total of 42 patients were admitted, including 18 males and 24 females. The onset age was (7.3±5.0) years, but the average age of diagnosis was (12.5±6.0) years. The lower limbs were involved in 38 cases, the upper limbs in 4 cases. There were 17 cases of stage Ⅰ, 17 cases of stage Ⅱ and 8 cases of stage Ⅲ. Only 4 cases had no misdiagnosis experience, and the misdiagnosis rate was 90.5%(38/42). Persistent pain, muscle contractures and joint deformities were the main clinical symptoms of the disease. MRI showed heterogeneous high and low signal intensity on T1-weighted images, and the high signal intensity was the same as that of subcutaneous adipose tissue. T2 fat-suppressed sequences showed stronger heterogeneous hyperintensity. The follow-up time was (14.6±10.8) months. Patients who took sirolimus orally before or after surgery experienced significant relief of pain symptoms. Of the 42 patients, the symptoms of 31 patients were completely relieved after the operation, and 11 patients still had residual pain or joint movement disorder or even deformity after the operation. Sixteen of 17 stage Ⅰ patients were cured, the lesion was further expanded and the pain recurred in 1 case after the operation. Of the 17 stage Ⅱ patients, 15 were cured, and 2 had mild limitation of ankle movement after the operation. Eight stage Ⅲ patients had varying degrees of pain or joint movement disorder after surgery, postoperative oral sirolimus significantly relieved symptoms. All 10 patients with stage Ⅰ and Ⅱ who underwent endoscopic resection were cured.Conclusion:FAVA usually occurred in school-age and adolescent children. Pain, muscle contracture and joint deformity were the characteristic clinical features. Combined with MRI features, the diagnosis can be confirmed. FAVA staging system could guide treatment and predict prognosis. For stage Ⅰ and Ⅱ patients, surgery should be performed as soon as possible and the prognosis was good. For stage Ⅲ patients, surgery-based comprehensive treatment could improve symptoms, but it was difficult to cure them radically. Oral sirolimus was also required after surgery.
6.Comparing the effect of different concentrations of contrast media in CT enterography
Yu LIU ; Jiuhong GUO ; Xiaoqing DAI ; Weiqing GAO ; Weilong DING ; Ya CHEN ; Yingjie YAN ; Xiaofeng TAO
Chinese Journal of Radiology 2020;54(5):417-422
Objective:To explore the enhancement effect of different concentrations of contrast media on blood vessels and hollow organs in CT enterography.Methods:Sixty patients with CT enterography were enrolled from January to August 2019 at Shanghai Ninth People's Hospital, and were prospectively randomly divided into three groups (group A: 90 ml 400 mg/ml contrast media, group B: 90 ml 350 mg/ml contrast media, and group C: 79 ml 400 mg/ml contrast media). Evaluation parameters included CT value, signal noise ratio (SNR) and contrast noise ratio (CNR) of main abdominal vessels (abdominal aorta, superior and inferior mesenteric atery, jejunal artery, ileum artery, superior and inferior mesenteric vein), jejunum, ileum and inflammatory bowel disease. The overall image quality and direct jejunum ileum artery quality scores were evaluated. One-way ANOVA was used to compare the parameters among the three groups.Results:There was no statistical difference in CT value, SNR and CNR of each measurement index among the three groups in plain scan ( P>0.05). The CT value, SNR and CNR of abdominal aorta, superior mesenteric artery, inferior mesenteric artery, jejunal artery and ileal artery in group A and C were higher than those in group B in arterial phase ( P<0.05), but there was no significant difference between group A and group C ( P>0.05). The CT value, SNR and CNR of superior mesenteric vein, inferior mesenteric vein, normal jejunum, ileum and inflammatory lesions in group A were higher than those in group B and C in venous phase ( P<0.05), but there was no significant difference between group B and group C ( P>0.05). The overall image quality scores of group A, B and C were (3.8±0.7), (3.4±0.6), (3.4±0.6), respectively, with no significant difference ( F=3.075, P=0.054). The direct jejunum ileum artery scores of the three groups were (3.5±0.5), (3.1±0.5), (3.4±0.5), respectively, the difference was statistically significant ( F=3.684, P=0.031). Conclusion:At the same injection rate and scanning parameters, contrast media at 400 mg/ml can provide better vessel and hollow organ enhancement.
7.Major depressive disorder with childhood trauma:Clinical characteristics, biological mechanism, and therapeutic implications.
Weilong GUO ; Jin LIU ; Lingjiang LI
Journal of Central South University(Medical Sciences) 2020;45(4):462-468
Major depressive disorder (MDD) is a main type of mood disorder, characterized by significant and lasting depressed mood. Until now, the pathogenesis of MDD is not clear, but it is certain that biological, psychological, and social factors are involved. Childhood trauma is considered to be an important factor in the development of this disease. Previous studies have found that nearly half of the patients with MDD have experienced childhood trauma, and different types of childhood trauma, gender, and age show different effects on this disease. In addition, the clinical characteristics of MDD patients with childhood trauma are also different, which often have more severe depressive symptoms, higher risk of suicide, and more severe cognitive impairment. The response to antidepressants is also worse. In terms of biological mechanisms and marker characteristics, the serotonin transporter gene and the FKBP prolyl isomerase 5 have been shown to play an important role in MDD and childhood trauma. Moreover, some brain imaging and biomarkers showed specific features, such as changes in gray matter in the dorsal lateral prefrontal cortex, and abnormal changes in hypothalamic-pituitary-adrenal axis function.
Child
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Depressive Disorder, Major
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Gray Matter
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Humans
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Hypothalamo-Hypophyseal System
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Pituitary-Adrenal System
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Suicide
8.A new butterfly femoral artery compression device vs manual compression for hemostasis of femoral artery puncture point after peripheral endovascular interventions
Weilong LU ; Bing WANG ; Ying WANG ; Heng XU ; Jinxuan PAN ; Mengyu WANG ; Peng GUO ; Yanling WANG ; Wenxuan SHANG
Journal of Interventional Radiology 2019;28(3):288-292
Objective To compare the hemostatic safety and efficacy of a new butterfly femoral artery compression device (FACD) with those of manual compression (MC) in patients undergoing percutaneous peripheral endovascular interventions via femoral artery. Methods A total of 283 patients, who received percutaneous endovascular interventions via femoral artery during the period from September 2016 to December 2017, were enrolled in this study. After endovascular intervention, 167 patients received FACD to make hemostasis (FACD group), and 116 patients received MC hemostasis (MC group) . The patient's comfortableness, time used for hemostasis (min), limb immobilization time (h), and the incidence of vascular complications in both groups were analyzed. Results All 283 patients were included in analysis, the results indicated that the hemostatic success rates in FACD group and MC group were 96.4% (161/167) and 94.0% (109/116) respectively, the difference between the two groups was not statistically significant (P>0.05) . The postoperative Kolcaba Comfort Scale score of FACD group was (85.0 ±11.2) points, which was remarkably higher than (58.4±11.7) points of MC group (P<0.05), the time used for hemostasis in FACD group was (9.2 ±2.2) min, which was strikingly shorter than (18.5 ±2.9) min in MC group (P <0.05) . The limb immobilization time in FACD group was (10.4±2.4) hours, which was obviously shorter than (23.1±4.1) hours in MC group (P <0.05) . The incidence of vascular complications in FACD group was 3.6%, which was dramatically lower than 9.5% in MC group (χ2=4.206, P=0.04) . Conclusion The use of the new butterfly FACD can promptly, safely and effectively stop bleeding of femoral artery puncture site. The new butterfly FACD is superior to MC in shortening hemostatic time and limb immobilization time, in reducing incidence of vascular complications, as well as in improving patient's comfortableness degree.
9.A new method for measuring pulmonary function parameters.
Jianming YANG ; Shenghe ZHONG ; Feixue LIANG ; Jinsong GUO ; Weilong LI ; Jinming LI
Journal of Southern Medical University 2013;33(9):1352-1356
OBJECTIVETo propose a new method for acquiring pulmonary function parameters based on measurement of volume changes of thoracic impedance.
METHODSWe studied the relationship between the volume changes of thoracic impedance and pulmonary function parameters during forced breathing based on bioimpedance measurement, and developed an instrument for measuring thoracic impedance. Using this instrument and a MRI spirolab III lung function test instrument, both based on flowmeter measurement, we measured such pulmonary function parameters including forced vital capacity (FVC), forced expiratory volume in one second/FVC (FEV1/FVC), and peak expiratory flow in 10 healthy volunteers and compared the measurement results.
RESULTSThe differences in the parameters measured using the two instruments were all within two folds of the positive and negative standard deviations of the average values, demonstrating good consistency in the measurement between the two methods.
CONCLUSIONSThe measurement results of the bioimpedance-based instrument we developed show good consistency with those by the commercially available pulmonary function test instrument.
Adult ; Electric Impedance ; Forced Expiratory Volume ; Humans ; Peak Expiratory Flow Rate ; Respiratory Function Tests ; methods ; Thorax ; physiology ; Vital Capacity ; Young Adult
10.Expression of human retinol-binding protein 4 in insect baculovirus system and preparation of its polyclonal antibody.
Yuying REN ; Dan CHEN ; Yuzheng GUO ; Hongna SHI ; Juan LIU ; Jingyang BAN ; Yaning LIU ; Xiaofang WU ; Weilong WANG ; Hai CHENG ; Dingfeng LI ; Yong LIU ; Liliang WANG
Chinese Journal of Biotechnology 2013;29(7):974-985
To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals
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Baculoviridae
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genetics
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Blotting, Western
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli
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Genetic Vectors
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Humans
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Immune Sera
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Insecta
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Rabbits
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Recombinant Proteins
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biosynthesis
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immunology
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Retinol-Binding Proteins, Plasma
;
biosynthesis
;
immunology
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Sf9 Cells
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metabolism
;
Transfection

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