1.Spheroid formation induced by human lung adenocarcinoma cell line SPC-A1 and the tumorigenic ability of lung cancer stem cells
Guangjun HAO ; Qing LIU ; Juan WANG ; Yane MA ; Yanhui DING
Chinese Journal of Tissue Engineering Research 2015;(23):3681-3685
BACKGROUND:There is no clear conclusion on whether the lung cancer stem cels can induce to spheroid formation and have tumorigenicity. OBJECTIVE: To observe the spheroid formation induced by human lung adenocarcinoma cel line SPC-A1 and the tumorigenic ability of lung cancer stem cels. METHODS:SPC-A1 at proliferating phase was cultured in serum-free DF12 culture medium, and then recombinant human insulin-like growth factor-1, recombinant human epidermal growth factor, and recombinant human fibroblast growth factor-10 were added to induce spheroid cels. Immune fluorescence detection and PCR amplification were done to understand the expression of stem cel associated markers. NOD-SCID immunodeficient mice were subcutaneously implanted with lung spheroid cels to observe the tumor growth.In vivo fluorescence imager was used for radiography. RESULTS AND CONCLUSION:After 5-10 days, lung spheroid cels were harvested. RT-PCR results showed that lung spheroid cels were positive for CD24, CD221, CCSP and SP-C. In addition, the lung spheroid cels and purified CD24+, CD221+ lung cancer stem cels were both positive for TTF-1 of lung stem cels, OCT4 and Nanog of embryonic stem cels and TTF-1 of Bmi-1 lung stem cels. The fluorescence detection showed that over 80% lung spheroid cels expressed CCSP and OCT4; SPC-A1 cels had the characteristics of alveolar type II cels, and also expressed SP-C protein, but only about 5% of the cels expressed CCSP and OCT4. At 50 days after subcutaneous implantation of lung spheroid cels, in vivo fluorescence imaging showed that the diameter of tumor in mice was 1 cm, indicating human lung adenocarcinoma cel line SPC-A1 can induce the spheroid formation, and lung cancer stem cels rich in the cel spheres have the tumorigenic ability.
2.Party for killing: the social behaviors in Cryptococcus neoformans.
Hao DING ; Guangjun HE ; Linqi WANG
Chinese Journal of Biotechnology 2017;33(9):1555-1566
Fungal pathogens represent an important group of human pathogenic microbes that lead to an unacceptably severe global burden especially due to exceptionally high mortality. For many fungal pathogens, they are widespread saprophytes and human host is not the exclusive niche for their proliferation. Their exceptional capability to survive and thrive within infected host likely stems from their sophisticated strategies in adaptation to diverse biotic and abiotic stressors from natural niches or predators. Among these 'environmental pathogens', Cryptococcus neoformans as a model organism claims the lives of more than half a million annually. Some recent studies indicate that cryptococcal survival both inside and outside of hosts can be coordinated by a combination of social behaviors. In this review, we describe and discuss the social behaviors employed by C. neoformans and address their significant impact on biofilm formation, sexual reproduction and pathogenicity.
3.Investigation and research on ticks carrying spotted fever group rickettsia in the border area of Tumen River Basin
Jixu LI ; Roh JONG?YUL ; Park WON?LL ; Wen PIAO ; Guangjun JIN ; Zhonggao WU ; Zhenhai SONG ; Shenghuan QUAN ; Guangming PIAO ; Hao SONG ; Cho SHIN?HYEONG
Chinese Journal of Preventive Medicine 2019;53(11):1130-1135
Objective To understand the situation and genotype distribution of spotted fever group rickettsia (SFGR) in the border area of Tumen River Basin in free ticks in Yanbian Korean Autonomous Prefecture (Yanbian Prefecture), Jilin Province. Methods From April to September, 2017, ticks were collected using flagging method from Hunchun, Tumen, Helong and Longjing cities in the Tumen River basin of Yanbian Prefecture. Outer membrane protein A (ompA) was detected by Polymerase Chain Reaction (PCR), then, the species were identified by gene sequencing and analyzed systematically. The positive rate of pools and MIR(minimum infection rate per 100 ticks,MIR) of SFGR were calculated, and the difference of positive rate of pools among ticks with different characteristics was compared by Chi?square test. Results A total of 3 079 ticks were collected and divided into 536 pools. The positive rate of pools of SFGR nucleic acid was 39.7% (213 pools). The MIR of SFGR was 6.9%.The positive rate of pools of SFGR in Dermacentor silvarum, Haemaphysalis concinna, Haemaphysalis japonica, Haemaphysalis longicornis and Ixodes persulcatus were 80.4% (41/51), 14.0% (25/179), 20.2% (18/89), 78.9% (101/128) and 25.9% (21/81), and the difference was statistically significant (P<0.001). There was statistical difference in the positive rate of pools of SFGR in developmental stages of ticks (P<0.001); the positive rate of pools of female adults, male adults, nymph and larvae were 36.4% (95/261), 34.2% (67/196), 56.3% (40/71) and 7/8, and the MIR was 7.9%, 7.7%, 4.9% and 3.5%. The five genotype was detected which was Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis, Candidatus Rickettsia tarasevichiae,Rickettsia monacensis and have 98%-100% homology with known gene sequences. Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis and Candidatus Rickettsia tarasevichiae showed close evolutionary relationship with known specie (have 98%-100% homology with known gene sequences); Rickettsia monacensis showed Far from evolutionary relationship with known species (have 98% homology with known gene sequences). Conclusion SFGR infection of ticks is common in the border areas of the Tumen River Basin. There was high diversity in SFGR species and tick species in the areas surveyed.
4.Investigation and research on ticks carrying spotted fever group rickettsia in the border area of Tumen River Basin
Jixu LI ; Roh JONG?YUL ; Park WON?LL ; Wen PIAO ; Guangjun JIN ; Zhonggao WU ; Zhenhai SONG ; Shenghuan QUAN ; Guangming PIAO ; Hao SONG ; Cho SHIN?HYEONG
Chinese Journal of Preventive Medicine 2019;53(11):1130-1135
Objective To understand the situation and genotype distribution of spotted fever group rickettsia (SFGR) in the border area of Tumen River Basin in free ticks in Yanbian Korean Autonomous Prefecture (Yanbian Prefecture), Jilin Province. Methods From April to September, 2017, ticks were collected using flagging method from Hunchun, Tumen, Helong and Longjing cities in the Tumen River basin of Yanbian Prefecture. Outer membrane protein A (ompA) was detected by Polymerase Chain Reaction (PCR), then, the species were identified by gene sequencing and analyzed systematically. The positive rate of pools and MIR(minimum infection rate per 100 ticks,MIR) of SFGR were calculated, and the difference of positive rate of pools among ticks with different characteristics was compared by Chi?square test. Results A total of 3 079 ticks were collected and divided into 536 pools. The positive rate of pools of SFGR nucleic acid was 39.7% (213 pools). The MIR of SFGR was 6.9%.The positive rate of pools of SFGR in Dermacentor silvarum, Haemaphysalis concinna, Haemaphysalis japonica, Haemaphysalis longicornis and Ixodes persulcatus were 80.4% (41/51), 14.0% (25/179), 20.2% (18/89), 78.9% (101/128) and 25.9% (21/81), and the difference was statistically significant (P<0.001). There was statistical difference in the positive rate of pools of SFGR in developmental stages of ticks (P<0.001); the positive rate of pools of female adults, male adults, nymph and larvae were 36.4% (95/261), 34.2% (67/196), 56.3% (40/71) and 7/8, and the MIR was 7.9%, 7.7%, 4.9% and 3.5%. The five genotype was detected which was Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis, Candidatus Rickettsia tarasevichiae,Rickettsia monacensis and have 98%-100% homology with known gene sequences. Candidatus Rickettsia longicornii, Rickettsia raoultii, Rickettsia heilongjiangensis and Candidatus Rickettsia tarasevichiae showed close evolutionary relationship with known specie (have 98%-100% homology with known gene sequences); Rickettsia monacensis showed Far from evolutionary relationship with known species (have 98% homology with known gene sequences). Conclusion SFGR infection of ticks is common in the border areas of the Tumen River Basin. There was high diversity in SFGR species and tick species in the areas surveyed.
5.A prospective randomized controlled study of antibiotic bone cement in the treatment of diabetic foot ulcer
Tao CAO ; Peng JI ; Zhi ZHANG ; Dan XIAO ; Kejia WANG ; Na LI ; Wen LI ; Guangjun JIN ; Tong HAO ; Ke TAO
Chinese Journal of Burns 2023;39(4):311-318
Objective:To investigate the clinical effects and related mechanism of antibiotic bone cement in treating diabetic foot ulcer (DFU).Methods:A prospective randomized controlled study was conducted. From August 2020 to August 2022, 24 patients with DFU who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University. According to the block randomization, the patients were divided into 2 groups, with 12 patients in each group. In antibiotic bone cement group, there were 7 male and 5 female patients, aged (64±8) years, with the ulcer area of (41±21) cm 2. In silver sulfadiazine group, there were 8 male and 4 female patients, aged (62±8) years, with the ulcer area of (38±19) cm 2. Under the condition of ensuring the patency of at least one main inferior genicular artery in each patient, the continuous vacuum sealing drainage was performed for 3-5 days after thorough debridement. Thereafter, the wounds in antibiotic bone cement group were treated with gentamicin-laden bone cement, and the wounds in silver sulfadiazine group were treated with silver sulfadiazine cream for dressing change. After 3 weeks of dressing change, the wound was covered with split-thickness skin graft from the lateral thigh on the affected side. Before debridement and after 3 weeks of dressing change, the blood flow intensities of wound tissue and normal skin tissue in foot were measured using laser Doppler flowmeter, and then, the percentage of relative blood flow intensity of wound and the change rate of blood flow intensity were calculated. After 3 weeks of dressing change, the wound margin tissue was taken, the number of CD31-positive neovascular and the vascular morphology were observed and detected by immunohistochemical staining, the morphology of blood vessels surrounded by CD31 and α-smooth muscle actin (α-SMA) double-positive cells was observed by immunofluorescence staining, the cell proliferation activity was evaluated by immunofluorescence staining (denoted as the ratio of Ki67 positive cells), and the protein expression of vascular endothelial growth factor receptor 2 (VEGFR2) was detected by Western blotting. The skin graft survival was observed 3-5 days after skin grafting, and the wound healing time was recorded. Data were statistically analyzed with independent sample t test and Fisher's exact probability test. Results:The percentages of relative blood flow intensity of wounds of patients before debridement were similar between the two groups ( P>0.05). After 3 weeks of dressing change, the percentage of relative blood flow intensity of wounds and the change rate of blood flow intensity of patients in antibiotic bone cement group were (44.7±2.0)% and (129±12)%, respectively, which were significantly higher than (28.3±1.2)% and (41±8)% in silver sulfadiazine group (with t values of 24.15 and 20.97, respectively, P<0.05). After 3 weeks of dressing change, compared with those in silver sulfadiazine group, the number of CD31-positive neovascular in the wound margin tissue of patients in antibiotic bone cement group was significantly increased ( t=33.81, P<0.05) with larger diameter and more regular arrangement, the vascular wall continuity surrounded by CD31 and α-SMA double-positive cells was better, and the ratio of Ki67 positive cells and protein expression of VEGFR2 were significantly increased (with t values of 40.97 and 47.38, respectively, P<0.05). On post skin grafting day 3-5, all the patients in antibiotic bone cement group and 8 patients in silver sulfadiazine group had good skin graft survival, while 4 patients in silver sulfadiazine group showed spotted/patchy skin graft necrosis, which were cured after corresponding treatment. The wound healing time of patients in antibiotic bone cement group was (47.1±2.9) d, which was significantly shorter than (58.8±2.3) d in silver sulfadiazine group ( t=10.86, P<0.05). Conclusions:Compared with silver sulfadiazine, clinical application of antibiotic bone cement for treating DFU has the characteristics of accelerating wound healing and better reconstruction of local blood flow, which may be closely related to the fact that antibiotic bone cement promoted the local angiogenesis effectively in the wound through enhancing the expression of VEGFR2.
6. Investigation and research on ticks carrying spotted fever group rickettsia in the border area of Tumen River Basin
Jixu LI ; Roh JONG-YUL ; Park WON-LL ; Wen PIAO ; Guangjun JIN ; Zhonggao WU ; Zhenhai SONG ; Shenghuan QUAN ; Guangming PIAO ; Hao SONG ; Cho SHIN-HYEONG
Chinese Journal of Preventive Medicine 2019;53(11):1130-1135
Objective:
To understand the situation and genotype distribution of spotted fever group rickettsia (SFGR) in the border area of Tumen River Basin in free ticks in Yanbian Korean Autonomous Prefecture (Yanbian Prefecture), Jilin Province.
Methods:
From April to September, 2017, ticks were collected using flagging method from Hunchun, Tumen, Helong and Longjing cities in the Tumen River basin of Yanbian Prefecture.