1.Development of a miniaturized CRISPR/Cas gene editing tool for human gut Bacteroides.
Yiwen ZHAN ; Linggang ZHENG ; Juntao SHEN ; Yucan HU ; Xuegang LUO ; Lei DAI
Chinese Journal of Biotechnology 2025;41(6):2360-2372
: Bacteroides, as one of the most abundant and diverse genera in the human gut, is regarded as a window into the study of gut microbiota-host interactions. Currently, CRISPR/Cas-based gene editing systems targeting Bacteroides have been widely applied, while the large size of Cas nucleases limits their potential application scenarios (such as in situ gut Bacteroides editing based on phage delivery). Therefore, this study aims to develop a compact and highly efficient genetic editing tool in Bacteroides., We developed a miniaturized CRISPR/Cas gene editing system for human gut Bacteroides. First, the editing capabilities of different miniaturized CRISPR/Cas systems, including AsCas12f, CasΦ2, and ISDge10, were evaluated in Bacteroides fragilis. Subsequently, the editing capability of AsCas12f was assessed across various Bacteroides species, and the size of this system was further optimized. The results demonstrated that the CRISPR/AsCas12f genome editing system exhibited the highest editing efficiency in B. fragilis. The CRISPR/AsCas12f system achieved efficient genome editing in B. fragilis, Bacteroides thetaiotaomicron, and Phocaeicola vulgatus. Furthermore, with a repair template of 500 bp homologous arms, the editing efficiency remained as high as 94.7%. In conclusion, CRISPR/AsCas12f can serve as a chassis tool enzyme for the development of Bacteroides-based miniature gene editors and derivative technologies, laying a foundation for the further development of gene editing technology for Bacteroides.
CRISPR-Cas Systems/genetics*
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Gene Editing/methods*
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Bacteroides/genetics*
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Humans
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Gastrointestinal Microbiome/genetics*
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Bacteroides fragilis/genetics*
2.Clinical effect of unilateral superior gluteal artery perforator propeller flap combined with contralateral centripetal advancement flap in repairing huge pressure ulcers in the sacrococcygeal region
Jun LI ; Mengdong LIU ; Liang ZHU ; Yuming LUO ; Qiying YANG ; Xiaowen GAO ; Juntao HAN
Chinese Journal of Burns 2024;40(6):551-556
Objective:To explore the feasibility and clinical effects of unilateral superior gluteal artery perforator propeller flap combined with contralateral centripetal advancement flap in repairing huge pressure ulcers in the sacrococcygeal region.Methods:The study was a retrospective observational study. From June 2020 to April 2023, 15 patients with stage Ⅳ pressure ulcers with sacrococcygeal defect area greater than 10.0 cm×10.0 cm who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University, including 8 males and 7 females, aged from 30 to 86 years. The pressure ulcers before debridement were all accompanied by different degree of infection and necrosis. Debridement and negative pressure sealing and irrigation treatment were performed in stage Ⅰ. After debridement, the skin and soft tissue defect area was 12.0 cm×10.5 cm to 20.0 cm×17.0 cm. After the wound bed infection was controlled, unilateral superior gluteal artery perforator propeller flap combined with contralateral centripetal advancement flap was used to repair the pressure ulcer wounds in stage Ⅱ. The perforator flap area was 12.0 cm×7.0 cm to 16.0 cm×10.5 cm. The donor area wound was sutured directly. After operation, the survival, complications, and wound healing of flap donor area were observed. During regular follow-up, the recurrence of pressure ulcers, the appearance and texture of the flap, and the scars in the donor site were observed.Results:After operation, 1 patient had fluid accumulation under the flap and survived after drainage and dressing change. The flaps of the other patients survived well without infection, local necrosis, and sinus formation under the flap. The wounds in the donor area healed well. All patients were followed up for more than 6 months, and there was no recurrence of pressure ulcers. The appearance of the flap was not bloated, the texture was soft, and the compression resistance and elasticity were good. The donor site wound healed well without obvious scar.Conclusions:The surgical method of repairing giant sacrococcygeal pressure ulcers with unilateral superior gluteal artery perforator propeller flap combined with contralateral centripetal advancement flap is simple and easy to operate. It can repair large defect area with the donor area being sutured directly, which is worthy of clinical promotion.
3.Numerical Simulation of Biomechanics of Two Aortic Root Replacement Procedures with Valve Preservation
Qian WANG ; Xinjin LUO ; Juntao QIU ; Shengyi HU ; Xuechao DING ; Tianming DU ; Yanping ZHANG ; Qianwen HOU ; Aike QIAO
Journal of Medical Biomechanics 2024;39(4):691-698
Objective To elucidate the influence of two procedures aortic root remodeling using a straight tubular artificial vessel while preserving the aortic valve and the Florida sleeve procedure on the biomechanics of the aortic root.Methods Five finite element models of the aortic root were reconstructed using computed tomography angiography images,including two cases of aortic root remodeling(A1 and A2),two cases of the Florida sleeve procedure(B1 and B2),and one control group without aortic root pathology(C).Numerical simulations were performed to obtain the blood flow and pressure distribution result to assess the differences in the hemodynamics of the aortic root.Results There were no significant differences in the peak systolic velocity between the two procedures and the control.However,the flow velocity after aortic root remodeling was smoother,similar to the model of the control group,with a more stable average aortic pressure and wall shear stress.In the Florida sleeve procedure,high-speed blood flow affected the vessel wall,leading to various degrees of wall shear stress and pressure concentrations along the aortic wall.Conclusions After aortic root replacement with valve preservation,blood flow patterns in the reconstructed aortic root depended on postoperative changes in sinus geometry.Both surgical procedures showed favorable blood flow patterns;however,the flow pattern after aortic root remodeling was more stable than that after the Florida sleeve procedure.
4.A Self-Developed Mobility Augmented Reality System Versus Conventional X-rays for Spine Positioning in Intraspinal Tumor Surgery: A Case-Control Study
Wenyao HONG ; Xiaohua HUANG ; Tian LI ; Juntao LUO ; Yuqing LIU ; Shengyue HUANG ; Zhongyi CHEN ; Bingwei HE ; Yuxing WEN ; Yuanxiang LIN
Neurospine 2024;21(3):984-993
Objective:
To evaluate the efficacy of a self-developed mobile augmented reality navigation system (MARNS) in guiding spinal level positioning during intraspinal tumor surgery based on a dual-error theory.
Methods:
This retrospective study enrolled patients diagnosed with intraspinal tumors admitted to Fujian Provincial Hospital between May and November 2023. The participants were divided into conventional x-rays and self-developed MARNS groups according to the localization methods they received. Position time, length of intraoperative incision variation, and location accuracy were systematically compared.
Results:
A total of 41 patients (19 males) with intraspinal tumors were included, and MARNS was applied to 21 patients. MARNS achieved successful lesion localization in all patients with an error of 0.38±0.12 cm. Compared to x-rays, MARNS significantly reduced positioning time (129.00±13.03 seconds vs. 365.00±60.43 seconds, p<0.001) and length of intraoperative incision variation (0.14 cm vs. 0.67 cm, p=0.009).
Conclusion
The self-developed MARNS, based on augmented reality technology for lesion visualization and perpendicular projection, offers a radiation-free complement to conventional x-rays.
5.A Self-Developed Mobility Augmented Reality System Versus Conventional X-rays for Spine Positioning in Intraspinal Tumor Surgery: A Case-Control Study
Wenyao HONG ; Xiaohua HUANG ; Tian LI ; Juntao LUO ; Yuqing LIU ; Shengyue HUANG ; Zhongyi CHEN ; Bingwei HE ; Yuxing WEN ; Yuanxiang LIN
Neurospine 2024;21(3):984-993
Objective:
To evaluate the efficacy of a self-developed mobile augmented reality navigation system (MARNS) in guiding spinal level positioning during intraspinal tumor surgery based on a dual-error theory.
Methods:
This retrospective study enrolled patients diagnosed with intraspinal tumors admitted to Fujian Provincial Hospital between May and November 2023. The participants were divided into conventional x-rays and self-developed MARNS groups according to the localization methods they received. Position time, length of intraoperative incision variation, and location accuracy were systematically compared.
Results:
A total of 41 patients (19 males) with intraspinal tumors were included, and MARNS was applied to 21 patients. MARNS achieved successful lesion localization in all patients with an error of 0.38±0.12 cm. Compared to x-rays, MARNS significantly reduced positioning time (129.00±13.03 seconds vs. 365.00±60.43 seconds, p<0.001) and length of intraoperative incision variation (0.14 cm vs. 0.67 cm, p=0.009).
Conclusion
The self-developed MARNS, based on augmented reality technology for lesion visualization and perpendicular projection, offers a radiation-free complement to conventional x-rays.
6.A Self-Developed Mobility Augmented Reality System Versus Conventional X-rays for Spine Positioning in Intraspinal Tumor Surgery: A Case-Control Study
Wenyao HONG ; Xiaohua HUANG ; Tian LI ; Juntao LUO ; Yuqing LIU ; Shengyue HUANG ; Zhongyi CHEN ; Bingwei HE ; Yuxing WEN ; Yuanxiang LIN
Neurospine 2024;21(3):984-993
Objective:
To evaluate the efficacy of a self-developed mobile augmented reality navigation system (MARNS) in guiding spinal level positioning during intraspinal tumor surgery based on a dual-error theory.
Methods:
This retrospective study enrolled patients diagnosed with intraspinal tumors admitted to Fujian Provincial Hospital between May and November 2023. The participants were divided into conventional x-rays and self-developed MARNS groups according to the localization methods they received. Position time, length of intraoperative incision variation, and location accuracy were systematically compared.
Results:
A total of 41 patients (19 males) with intraspinal tumors were included, and MARNS was applied to 21 patients. MARNS achieved successful lesion localization in all patients with an error of 0.38±0.12 cm. Compared to x-rays, MARNS significantly reduced positioning time (129.00±13.03 seconds vs. 365.00±60.43 seconds, p<0.001) and length of intraoperative incision variation (0.14 cm vs. 0.67 cm, p=0.009).
Conclusion
The self-developed MARNS, based on augmented reality technology for lesion visualization and perpendicular projection, offers a radiation-free complement to conventional x-rays.
7.A Self-Developed Mobility Augmented Reality System Versus Conventional X-rays for Spine Positioning in Intraspinal Tumor Surgery: A Case-Control Study
Wenyao HONG ; Xiaohua HUANG ; Tian LI ; Juntao LUO ; Yuqing LIU ; Shengyue HUANG ; Zhongyi CHEN ; Bingwei HE ; Yuxing WEN ; Yuanxiang LIN
Neurospine 2024;21(3):984-993
Objective:
To evaluate the efficacy of a self-developed mobile augmented reality navigation system (MARNS) in guiding spinal level positioning during intraspinal tumor surgery based on a dual-error theory.
Methods:
This retrospective study enrolled patients diagnosed with intraspinal tumors admitted to Fujian Provincial Hospital between May and November 2023. The participants were divided into conventional x-rays and self-developed MARNS groups according to the localization methods they received. Position time, length of intraoperative incision variation, and location accuracy were systematically compared.
Results:
A total of 41 patients (19 males) with intraspinal tumors were included, and MARNS was applied to 21 patients. MARNS achieved successful lesion localization in all patients with an error of 0.38±0.12 cm. Compared to x-rays, MARNS significantly reduced positioning time (129.00±13.03 seconds vs. 365.00±60.43 seconds, p<0.001) and length of intraoperative incision variation (0.14 cm vs. 0.67 cm, p=0.009).
Conclusion
The self-developed MARNS, based on augmented reality technology for lesion visualization and perpendicular projection, offers a radiation-free complement to conventional x-rays.
8.A Self-Developed Mobility Augmented Reality System Versus Conventional X-rays for Spine Positioning in Intraspinal Tumor Surgery: A Case-Control Study
Wenyao HONG ; Xiaohua HUANG ; Tian LI ; Juntao LUO ; Yuqing LIU ; Shengyue HUANG ; Zhongyi CHEN ; Bingwei HE ; Yuxing WEN ; Yuanxiang LIN
Neurospine 2024;21(3):984-993
Objective:
To evaluate the efficacy of a self-developed mobile augmented reality navigation system (MARNS) in guiding spinal level positioning during intraspinal tumor surgery based on a dual-error theory.
Methods:
This retrospective study enrolled patients diagnosed with intraspinal tumors admitted to Fujian Provincial Hospital between May and November 2023. The participants were divided into conventional x-rays and self-developed MARNS groups according to the localization methods they received. Position time, length of intraoperative incision variation, and location accuracy were systematically compared.
Results:
A total of 41 patients (19 males) with intraspinal tumors were included, and MARNS was applied to 21 patients. MARNS achieved successful lesion localization in all patients with an error of 0.38±0.12 cm. Compared to x-rays, MARNS significantly reduced positioning time (129.00±13.03 seconds vs. 365.00±60.43 seconds, p<0.001) and length of intraoperative incision variation (0.14 cm vs. 0.67 cm, p=0.009).
Conclusion
The self-developed MARNS, based on augmented reality technology for lesion visualization and perpendicular projection, offers a radiation-free complement to conventional x-rays.
9.Hepatic DDAH1 mitigates hepatic steatosis and insulin resistance in obese mice: Involvement of reduced S100A11 expression.
Xiyue SHEN ; Kai LUO ; Juntao YUAN ; Junling GAO ; Bingqing CUI ; Zhuoran YU ; Zhongbing LU
Acta Pharmaceutica Sinica B 2023;13(8):3352-3364
Dimethylarginine dimethylaminohydrolase 1 (DDAH1) is an important regulator of plasma asymmetric dimethylarginine (ADMA) levels, which are associated with insulin resistance in patients with nonalcoholic fatty liver disease (NAFLD). To elucidate the role of hepatic DDAH1 in the pathogenesis of NAFLD, we used hepatocyte-specific Ddah1-knockout mice (Ddah1HKO) to examine the progress of high-fat diet (HFD)-induced NAFLD. Compared to diet-matched flox/flox littermates (Ddah1f/f), Ddah1HKO mice exhibited higher serum ADMA levels. After HFD feeding for 16 weeks, Ddah1HKO mice developed more severe liver steatosis and worse insulin resistance than Ddah1f/f mice. On the contrary, overexpression of DDAH1 attenuated the NAFLD-like phenotype in HFD-fed mice and ob/ob mice. RNA-seq analysis showed that DDAH1 affects NF-κB signaling, lipid metabolic processes, and immune system processes in fatty livers. Furthermore, DDAH1 reduces S100 calcium-binding protein A11 (S100A11) possibly via NF-κB, JNK and oxidative stress-dependent manner in fatty livers. Knockdown of hepatic S100a11 by an AAV8-shS100a11 vector alleviated hepatic steatosis and insulin resistance in HFD-fed Ddah1HKO mice. In summary, our results suggested that the liver DDAH1/S100A11 axis has a marked effect on liver lipid metabolism in obese mice. Strategies to increase liver DDAH1 activity or decrease S100A11 expression could be a valuable approach for NAFLD therapy.
10.Outcomes and preoperative evaluation of transfemoral transcatheter aortic valve replacement (TAVR) in the treatment of pure native aortic valve regurgitation
Donghui XU ; Xinjin LUO ; Xu WANG ; Yuetang WANG ; Xiang FENG ; Xuanshu LI ; Juntao QIU ; Wei WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2021;28(09):1054-1060
Objective To assess outcomes of transcatheter aortic valve replacement (TAVR) for pure native aortic valve regurgitation. Methods A total of 129 patients underwent transfemoral TAVR in Fuwai Hospital from May 2019 to October 2020 were retrospectively analyzed. There were 83 males and 46 females with an average age of 72.26±8.97 years. The patients were divided into a pure native aortic valve regurgitation group (17 patients) and an aortic valve stenosis group (112 patients). Results The incidence of valve in valve was higher in the pure native aortic valve regurgitation group (47.0% vs. 16.1%, P<0.01). There was no statistical difference between the two groups in conversion to surgery, intraoperative use of extracorporeal circulation, intraoperative left ventricular rupture, postoperative use of extracorporeal membrane oxygenation (ECMO), peripheral vascular complications, disabled stroke, death, or pacemaker implantation. There was no statistical difference in the diameter of annulus (25.75±2.21 mm vs. 24.70±2.90 mm, P=0.068) or diameter of outflow tract (25.82±3.75 mm vs. 25.37±3.92 mm, P=0.514) between the pure native aortic valve regurgitation group and the aortic valve stenosis group. Conclusion Transfemoral TAVR is a feasible method for patients with pure native aortic valve regurgitation. The diameter of annulus plane, the diameter of outflow tract and the shape of outflow tract should be evaluated.

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