Transplantation of Gelatin Microspheres Loaded with Wharton's Jelly Derived Mesenchymal Stem Cells Facilitates Cartilage Repair in Mice

Chen Xiaolin,Huang Sunxing,Niu Yongxia,Luo Mingxun,Liu Haiying,Jiao Yiren,Huang Junjiu 한국조직공학과 재생의학회 2024 조직공학과 재생의학 Vol.21 No.1

Background: Knee osteoarthritis (KOA) is a prevalent chronic joint disease caused by various factors. Mesenchymal stem cells (MSCs) therapy is an increasingly promising therapeutic option for osteoarthritis. However, the chronic inflammation of knee joint can severely impede the therapeutic effects of transplanted cells. Gelatin microspheres (GMs) are degradable biomaterial that have various porosities for cell adhesion and cell–cell interaction. Excellent elasticity and deformability of GMs make it an excellent injectable vehicle for cell delivery. Methods: We created Wharton’s jelly derived mesenchymal stem cells (WJMSCs)-GMs complexes and assessed the effects of GMs on cell activity, proliferation and chondrogenesis. Then, WJMSCs loaded in GMs were transplanted in the joint of osteoarthritis mice. After four weeks, joint tissue was collected for histological analysis. Overexpressing-luciferase WJMSCs were performed to explore cell retention in mice. Results: In vitro experiments demonstrated that WJMSCs loaded with GMs maintained cell viability and proliferative potential. Moreover, GMs enhanced the chondrogenesis differentiation of WJMSCs while alleviated cell hypertrophy. In KOA mice model, transplantation of WJMSCs-GMs complexes promoted cartilage regeneration and cartilage matrix formation, contributing to the treatment of KOA. Compared with other groups, in WJMSCs+GMs group, there were fewer cartilage defects and with a more integrated tibia structure. Tracking results of stable-overexpressing luciferase WJMSCs demonstrated that GMs significantly extended the retention time of WJMSCs in knee joint cavity. Conclusion: Our results indicated that GMs facilitate WJMSCs mediated knee osteoarthritis healing in mice by promoting cartilage regeneration and prolonging cell retention. It might potentially provide an optimal strategy for the biomaterial-stem cell based therapy for knee osteoarthritis. Background: Knee osteoarthritis (KOA) is a prevalent chronic joint disease caused by various factors. Mesenchymal stem cells (MSCs) therapy is an increasingly promising therapeutic option for osteoarthritis. However, the chronic inflammation of knee joint can severely impede the therapeutic effects of transplanted cells. Gelatin microspheres (GMs) are degradable biomaterial that have various porosities for cell adhesion and cell–cell interaction. Excellent elasticity and deformability of GMs make it an excellent injectable vehicle for cell delivery. Methods: We created Wharton’s jelly derived mesenchymal stem cells (WJMSCs)-GMs complexes and assessed the effects of GMs on cell activity, proliferation and chondrogenesis. Then, WJMSCs loaded in GMs were transplanted in the joint of osteoarthritis mice. After four weeks, joint tissue was collected for histological analysis. Overexpressing-luciferase WJMSCs were performed to explore cell retention in mice. Results: In vitro experiments demonstrated that WJMSCs loaded with GMs maintained cell viability and proliferative potential. Moreover, GMs enhanced the chondrogenesis differentiation of WJMSCs while alleviated cell hypertrophy. In KOA mice model, transplantation of WJMSCs-GMs complexes promoted cartilage regeneration and cartilage matrix formation, contributing to the treatment of KOA. Compared with other groups, in WJMSCs+GMs group, there were fewer cartilage defects and with a more integrated tibia structure. Tracking results of stable-overexpressing luciferase WJMSCs demonstrated that GMs significantly extended the retention time of WJMSCs in knee joint cavity. Conclusion: Our results indicated that GMs facilitate WJMSCs mediated knee osteoarthritis healing in mice by promoting cartilage regeneration and prolonging cell retention. It might potentially provide an optimal strategy for the biomaterial-stem cell based therapy for knee osteoarthritis.

Effect of Laminated Structure on the Mechanical Properties of Reclaimed Bamboo Chopsticks-Wood Veneer Hybrid Laminated Composite

Haiying Zhou,Xin Wei,Fuming Chen,Ge Wang,Lee M. Smith 한국섬유공학회 2019 Fibers and polymers Vol.20 No.7

The waste created by disposable bamboo chopsticks used in dine in restaurants, fast food, and takeaway has anincreasingly enormous impact. The object of this study was to investigate the effect of the type of laminate structure on themechanical properties of a novel type of biodegradable bamboo-wood hybrid composite processed from the reclaimedbamboo chopsticks and wood veneer hybrid laminate composite (BWHC). Four types of laminate structures were chosen forcomparison namely: bamboo chopsticks parallel laminate (Type I), bamboo chopsticks cross-laminate (Type II), bamboochopsticks-wood veneer parallel laminate (Type III), and bamboo chopsticks-wood veneer cross-laminate (Type IV). Theresults revealed that the modulus of rupture MOR of Type I>Type III>Type IV>Type II. The optimum bonding strength forBWHC was displayed by the structural Type III, followed by Types I, IV and II. The horizontal wood veneers layer betweenthe bamboo chopsticks layers effectively improve the bonding strength of BWHC which was confirmed by SEM observation. The dynamic mechanical properties of BWHC were examined by performing low velocity impact loading tests. The impactperformance of BWHC was significantly greater in the samples designed with a cross-laminate structure. Mixed failuremodels for the BWHC under the impact test were found, including debonding, delamination, fiber tensile fracture, andstructural collapse. Structural collapse was main failure model of BWHC with a cross hybrid lamination and their values ofthe total energy absorbed was far greater than BWHC with parallel laminate. This study suggest that bamboo-wood hybridcomposite with low cost, biodegradability, and excellent physical and mechanical properties can be manufactured by therecycling utilization and structural design.

Whole-genome identification and salt- and ABA-induced expression trends of the Nicotiana tabacum CKX gene family

Chen Wenwen,Wang Genhong,Yi Meiqin,Gao Tianke,Song Chunman,Gao Qian,Xia Qingyou,Xiang Haiying 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.3

Cytokinin hormones are indispensable for plant growth and development. Cytokinin oxidase/dehydrogenase (CKX) helps regulate the dynamic balance of endogenous cytokinin levels. However, little is known about the CKX genes of Nicotiana tabacum (NtCKXs). In silico analyses were used to isolate, characterize, and elucidate the phylogenetic relationships of 15 NtCKX genes. Multi-species phylogenetic tree construction placed NtCKX1–15 on five of the eight branches of the CKX phylogenetic tree. Protein structure and network analyses revealed that NtCKX genes located on the same phylogenetic branch generally contained several conserved motifs and have highly similar structures, with structural domains related to flavin adenine dinucleotide (FAD) and cytokinin-binding found on all of the predicted NtCKX-encoded proteins. The upstream promotor region of NtCKX genes contained many abiotic stress-responsive cis-acting elements, including DRE, ERE, MBS, MYB, and MYC. Gene expression analysis revealed that each NtCKX gene responded differently to salt stress and exogenous abscisic acid (ABA) treatment. Four NtCKX genes exhibited ABA-induced expression trends with varying peak times. Under salt stress, NtCKX expression was significantly suppressed in two genes and upregulated in five others. In summary, we provided basic information regarding the CKX gene family of N. tabacum and elucidated their gene expression patterns under abiotic stresses, including ABA treatment and salt stress. The findings of this work can serve as a foundation for future study of the functions of NtCKX genes.

Effects of rumen-protected biotin on the growth performance, nitrogen utilization and blood parameters of yearling Liaoning cashmere doelings

Liu Haiying,Lin Ying,Chen Xuhui,Yang Guiqin 아세아·태평양축산학회 2023 Animal Bioscience Vol.36 No.11

Objective: This study was conducted to investigate the effects of rumen-protected biotin (RPB) on growth performance, nutrient digestibility, nitrogen utilization and plasma biochemical parameters of Liaoning cashmere goats during the cashmere fiber growing period. Methods: Sixteen 6-month-old Liaoning cashmere twin-doelings (24.8±1.20 kg) were allocated to 2 diet groups that were individually ad libitum fed 30% concentrate and 70% forage diet (dry matter [DM]) by a paired experimental design. Goats of the control group were fed the basal diet, while goats belonging to the RPB group were fed the basal diet with 10 mg RPB/d per animal. The duration of the experiment was 16 weeks with two 8-week periods. Digestibility was determined at weeks 7 and 15, and other measures were taken every four weeks. Results: Compared with the control group, the average daily gain of the RPB group increased by 10.94% (p<0.05), and the intake of neutral detergent fiber was increased (p = 0.045). There were some increasing tendencies for the intake of DM, acid detergent fiber and ether extract (p = 0.070, 0.088, and 0.070, respectively). The intake and digestibility of N tended to increase (p = 0.062 and 0.093, respectively), while the N fecal excretion percentage of N intake was decreased (p = 0.093) in the RPB compared with the control group. N retention tended to increase (p = 0.084) with the addition of adding RPB to the diet. Plasma total protein was increased (p = 0.037), whereas the urea-N concentration was decreased (p = 0.049) in the RPB diet group compared with the control diet group. The levels of propionyl-CoA carboxylase (p<0.001) and methylmalonyl-CoA (p = 0.013) were increased in the RPB group. Conclusion: Supplementation of rumen-protected biotin in the diet of cashmere goats can enhance the utilization of N and improve daily weight gain during cashmere fiber growing period.

Interaction between dietary digestible tryptophan and soy oligosaccharides in broiler chickens: effects on caecal skatole level and microflora

Jing Chen,Hansong Jing,Haiying Liu,Xin Zhu,Guiqin Yang Asian Australasian Association of Animal Productio 2023 Animal Bioscience Vol.36 No.3

Objective: This study was conducted to evaluate the interactive effects of dietary digestible tryptophan (dTry) and soy oligosaccharides (SO) on growth performance, caecal skatole level, and microflora of broiler chickens aged from 14 to 42 days. Methods: Three hundred and sixty broiler chicks were allocated equally to 36 cages at 14-day-of-age according to body weight and gender. Using a 3×2 factorial arrangement, 3 dietary dTry levels (0.18%, 0.23%, and 0.28%) supplemented with 0 or 3.5 g/kg of SO were used to create 6 diets (treatments). Each diet was fed to six replicates of 10 birds (60 birds/treatment), growth performance was measured. Caecal content samples were collected at 42 days of age. Results: Results showed that significantly different dTry level×SO interactions were found for average daily gain (ADG), caecal levels of indole, propionic acid, and butyric acid, and microbial Shannon index (p<0.05). Birds fed diet containing 0.23% dTry level with SO supplementation had higher ADG and lower feed/gain ratio than those fed the other diets (p<0.05). Broilers fed diets containing 0.28% dTry increased their caecal levels of indole and skatole compared with those containing 0.18% or 0.23% dTry (p<0.01), regardless of SO addition. SO supplementation to diets decreased the caecal skatole level by 16.17% (p<0.05), and increased the relative frequency of Clostridium IV (p<0.05), regardless of dietary dTry level. Conclusion: These results indicated that diets containing 0.23% dTry with SO supplementation positively promoted ADG, and decreased caecal skatole levels of broiler chickens. The dietary dTry level, SO affected the caecal skatole level, however, there was no interaction between them.

Severe Tick Infestation in a Hare and Potential Risk for Transmitting Pathogens to Humans

Weiqing Zheng,Haiying Chen,Xiaoqing Liu,Xuejian Guo,Renlong Fu 대한기생충학열대의학회 2011 The Korean Journal of Parasitology Vol.49 No.4

Cloning and sequence analysis of Wild Argali short palate, lung and nasal epithelium clone 1 cDNA

Wen Shen,Kaili Chen,Yanming Sun,Haiying Guo,Dongmei Chen,Yang Cao 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.5

Objective: Experiments were conducted to clone the sequence of Wild Argali short palate, lung and nasal epithelium clone 1 (SPLUNC1) cDNA, and to lay the foundation for further study the biological function of Wild Argali SPLUNC1. Methods: The complete sequence of Wild Argali SPLUNC1 cDNA was generated by rapid amplification of cDNA ends. The entire coding sequence was inserted into the pPIC9K vector and expressed in Pichia pastoris (P. pastoris) GS115. The recombinant SPLUNC1 protein was detected by Western blot and purified by Ni2+ chelate affinity chromatography. The test of effect of the protein on Mycoplasma ovipneumoniae (MO) was performed with real-time polymerase chain reaction. Results: The Wild Argali SPLUNC1 cDNA was 1,076 bp with an open reading frame of 768 bp, which encoded a 26.49 kDa protein composed of 255 amino acids. Its amino acid sequence shared 98.4%, 96.9%, 94.5%, 90.2%, 80.8%, 78.4%, 78.3%, 72.5%, 72.3%, 68.8% identity with those of SPLUNC1 cDNA from Ovis aries (accession no. NP_001288334.1), Capra hircus (accession no. XP_005688516.1), Pantholops hodgsonii (accession no. XP_005979709.1), Bos taurus (accession no. NP_776851.1), Felis catus (accession no. XP_006929910.1), Homo sapiens (accession no. NP_001230122.1), Sus scrofa (accession no. NP_001005727.1), Chinchilla lanigera (accession no. NP_001269294.1), Mus musculus (accession no. NP_035256.2), and Rattus norvegicus (accession no. NP_742028.1), respectively. The recombinant protein corresponded to the expected molecular mass of 25.47 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was detected in the supernatant of P. pastoris, and it could be purified. The results from the test of inhibition effect of argali recombinant SPLUNC1 protein on MO showed that the product could inhibit MO very well (p<0.01). Conclusion: The amino acid sequence of Wild Argali SPLUNC1 was different from other organisms. The recombinant SPLUNC1 protein has good biological activity.

Electromagnetic Analysis and Optimization for a Dual-Rotor Switched Reluctance Machine

Xing Wang,Rui Nie,Hao Chen,Haiying Wang,Tong Xu 한국자기학회 2019 Journal of Magnetics Vol.24 No.2

A dual-rotor switched reluctance motor is proposed in this paper. The structure and working principle of the motor are introduced in detail. The structural parameters have great influence on the performance of the motor, so sensitivity analysis of several important structural parameters to the output torque is carried out based on the two-dimensional finite element method, which improves the average torque of the rotor in the outer air gap. Additionally, the performance of the motor with various pole distributions is analyzed and compared. The results show that when the magnetic polarity distribution of the outer stator is NSNSNSNSNSNS and the magnetic polarity distribution of the inner rotor is NSNSNS, the motor has the best performance. The torque of the motor is compared with that of the traditional single-rotor switched reluctance motor. It shows that the performance of the motor proposed in this paper is better.

Vibrio parahaemolyticus cqsA controls production of quorum sensing signal molecule 3-hydroxyundecan-4-one and regulates colony morphology

Kui Wu,Yangyun Zheng,Qingping Wu,Haiying Chen,Songzhe Fu,Biao Kan,Yongyan Long,Xiansheng Ni,Junling Tu 한국미생물학회 2019 The journal of microbiology Vol.57 No.12

In order to adapt to different environments, Vibrio parahaemolyticus employed a complicated quorum sensing system to orchestrate gene expression and diverse colony morphology patterns. In this study, the function of the putative quorum sensing signal synthase gene cqsA (VPA0711 in V. parahaemolyticus strain RIMD2210633 genome) was investigated. The cloning and expression of V. parahaemolyticus cqsA in Escherichia coli system induced the production of a new quorum sensing signal that was found in its culture supernatant. The signal was purified by high performance liquid chromatography methods and determined to be 3-hydroxyundecan- 4-one by indirect and direct mass spectra assays. The deletion of cqsA in RIMD2210633 changed V. parahaemolyticus colony morphology from the classical ‘fried-egg’ shape (thick and opaque in the center, while thin and translucent in the edge) of the wild-type colony to a ‘pancake’ shape (no significant difference between the centre and the edge) of the cqsAdeleted colony. This morphological change could be restored by complementary experiment with cqsA gene or the signal extract. In addition, the expression of opaR, a well-known quorum sensing regulatory gene, could be up-regulated by cqsA deletion. Our results suggested that V. parahaemolyticus used cqsA to produce 3-hydroxyundecan-4-one signal and thereby regulated colony morphology and other quorum sensing-associated behaviors.

Description of Corynebacterium poyangense sp. nov., isolated from the feces of the greater white-fronted geese (Anser albifrons)

Liu Qian,Fan Guoying,Wu Kui,Bai Xiangning,Yang Xi,Song Wentao,Chen Shengen,Xiong Yanwen,Chen Haiying 한국미생물학회 2022 The journal of microbiology Vol.60 No.7

Two novel Gram-positive, non-spore-forming, facultatively anaerobic, non-motile, and short rods to coccoid strains were isolated from the feces of the greater white-fronted geese (Anser albifrons) at Poyang Lake. The 16S rRNA gene sequences of strains 4H37-19T and 3HC-13 shared highest identity to that of Corynebacterium uropygiale Iso10T (97.8%). Phylogenetic and phylogenomic analyses indicated that strains 4H37-19T and 3HC-13 formed an independent clade within genus Corynebacterium and clustered with Corynebacterium uropygiale Iso10T. The average nucleotide identity and digital DNA-DNA hybridization value between strains 4H37-19T and 3HC-13 and members within genus Corynebacterium were all below 95% and 70%, respectively. The genomic G + C content of strains 4H37-19T and 3HC-13 was 52.5%. Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidylcholine, and phosphatidyl inositol mannosides (PIM) were the major polar lipids, with C18:1ω9c, C16:0, and C18:0 as the major fatty acids, and MK-8 (H4), MK-8(H2), and MK-9(H2) as the predominant respiratory quinones. The major whole cell sugar was arabinose, and the cell wall included mycolic acids. The cell wall peptidoglycan contained meso-diaminopimelic acid (meso-DAP). The polyphasic taxonomic data shows that these two strains represent a novel species of the genus Corynebacterium, for which the name Corynebacterium poyangense sp. nov. is proposed. The type strain of Corynebacterium poyangense is 4H37-19T (=GDMCC 1.1738T = KACC 21671T).