1.Application of holographic imaging digital localization technology in robotic partial nephrectomy for completely endophytic renal tumor
Zhengsheng LIU ; Shuaishuai SONG ; Xuegang WANG ; Kaiyan ZHANG ; Wei LI ; Zhun WU ; Bin CHEN ; Tao WANG ; Jinchun XING
Chinese Journal of Urology 2025;46(5):376-382
Objective:To evaluate the clinical value of holographic imaging-based digital localization technology in robot-assisted partial nephrectomy(RAPN)for the treatment of completely endophytic renal tumors.Methods:A retrospective analysis was conducted on the clinical data of 23 patients with completely endophytic renal tumors who underwent RAPN at the First Affiliated Hospital of Xiamen University between December 2022 and December 2024. Patients were divided into two groups based on the use of holographic imaging:the holographic imaging group(16 cases)and the conventional group(7 cases). There was no statistically significant difference between the holographic imaging group and the conventional group in terms of age[(41.9 ± 13.4)years vs.(46.9 ± 13.4)years],body mass index[(25.6 ± 4.8)kg/m2 vs.(24.7 ± 3.1)kg/m2],maximum tumor diameter[(3.1 ± 0.9)cm vs.(3.0 ± 9.0)cm],tumor volume[(13.2 ± 9.0)cm3 vs.(34.9 ± 9.9)cm3],R.E.N.A.L. score[(9.4 ± 1.2)points vs.(9.9 ± 0.7)points],PADUA score[(10.4 ± 0.7)points vs.(9.4 ± 0.7)points],proportion of T 1a stage patients[12 cases(75.0%)vs. 6 cases(85.7%)]and preoperative serum creatinine[(67.4 ± 9.5)μmol/L vs.(78.0 ± 16.0)μmol/L]. In the holographic group,holographic models were reconstructed based on preoperative enhanced CT or MRI images and used for preoperative planning and intraoperative localization. In the conventional group,surgeons relied on preoperative CT or MRI images for cognitive fusion during RAPN. Perioperative parameters such as warm ischemia time,operative time,tumor localization time,positive surgical margin rate,and renal function were compared between the two groups. Results:The operative time in the holographic group was significantly shorter than that in the conventional group[(152.8 ± 12.9)min vs.(218.4 ± 105.5)min, P = 0.001]. Warm ischemia time[(26.9 ± 3.4)min vs.(30.7 ± 3.8)min, P < 0.001],localization time[(4.2 ± 0.9)min vs.(8.9 ± 1.7)min, P < 0.001],and estimated blood loss[(47.0 ± 17.7)ml vs.(128.6 ± 87.8)ml, P < 0.001]were also significantly lower in the holographic group. In the conventional group,one patient underwent radical nephrectomy,while no patient in the holographic imaging group required conversion to radical nephrectomy. No cases of positive surgical margins were identified in either group. Serum creatinine levels measured one month after surgery showed no statistically significant differences between the two groups[(79.5 ± 15.7)μmol /L vs.(104.9 ± 22.5)μmol /L]. Conclusions:The application of holographic imaging-based digital localization technology in RAPN for completely endophytic renal tumors significantly reduces operative time,localization time,warm ischemia time,and intraoperative blood loss. This technology improves surgical efficiency and success rates,offering distinct clinical advantages.
2.Expert consensus on local anesthesia application in pediatric dental therapies.
Yan WANG ; Jing ZOU ; Yang JI ; Jun WANG ; Bin XIA ; Wei ZHAO ; Li'an WU ; Guangtai SONG ; Yuan LIU ; Xu CHEN ; Jiajian SHANG ; Qin DU ; Qingyu GUO ; Beizhan JIANG ; Hongmei ZHANG ; Xianghui XING ; Yanhong LI
West China Journal of Stomatology 2025;43(4):455-461
Dental treatments for children and adolescents have unique clinical characteristics that differ from dental care for adults in terms of children's physiology, psychology, and behavior. These differences impose specific requirements on the application of local anesthesia in pediatric dental procedures. This article presents expert consensus on the principles of local anesthesia techniques in pediatric dental therapies, including the use of common anesthetic drugs and dosage control, safety and efficacy evaluation, and prevention and management of complications. The aim is to improve the safety and quality of pediatric dental treatments and offer guidance for clinical application by dentists.
Humans
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Child
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Anesthesia, Local/methods*
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Consensus
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Anesthesia, Dental/methods*
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Adolescent
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Anesthetics, Local/administration & dosage*
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Dental Care for Children
3.Effect of Modified Zhigancao Granules (炙甘草汤加味颗粒) on Early Recurrence Following Radiofrequency Ablation in Patients with Atrial Fibrillation of Qi-Yin Deficiency Syndrome:A Randomized,Double-Blind,Placebo-Controlled Trial
Yucai HU ; Boyong QIU ; Jingjing WEI ; Bin LI ; Zuoying XING ; Huixia PENG ; Mingjun ZHU ; Yongxia WANG
Journal of Traditional Chinese Medicine 2025;66(23):2460-2466
ObjectiveTo evaluate the clinical effectiveness and safety of Modified Zhigancao Granules (炙甘草汤加味颗粒) for preventing the early recurrence following radiofrequency ablation in patients with atrial fibrillation (AF) of qi-yin deficiency syndrome. MethodsA multi-center, randomized, double-blind, placebo-controlled trial was designed. A total of 116 patients with atrial fibrillation of qi-yin deficiency syndrome who underwent radiofrequency ablation for the first time were enrolled from 3 centers, and they were randomly divided into a treatment group (59 cases) and a control group (57 cases). Both groups received basic western medicine treatment after surgery. In addition, the treatment group was given oral Modified Zhigancao Granules, while the control group was given oral placebo granules. The dosage for both groups was 20 g each time, twice a day, with continuous treatment for 12 weeks. The recurrence of atrial fibrillation in both groups was recorded at 24 hours, 4 weeks, 8 weeks, and 12 weeks after surgery. The serum levels of B-type natriuretic peptide (BNP), interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor-α (TNF-α) were detected before treatment and 12 weeks after treatment in both groups. The scores of Atrial Fibrillation Effect on QualiTy-of-life (AFEQT) Questionnaire (including scores of the daily life dimension, symptom dimension, treatment worry dimension, treatment satisfaction dimension, and total score) and traditional Chinese medicine (TCM) syndrome scores were compared before treatment and at 4 weeks, 8 weeks, and 12 weeks between groups. Safety indicators such as blood routine, urine routine, liver function, and renal function were monitored before and after treatment. ResultsNine of the treatment group and seven of the control group dropped out. Finally, 50 patients in each group were included in the statistical analysis. At 24 hours, 4 weeks, 8 weeks, and 12 weeks after surgery, the recurrence rates of AF in the treatment group were 2.0% (1/50), 2.0% (1/50), 4.0% (2/50), and 10.0% (5/50), respectively; while those in the control group were 2.0% (1/50), 26.0% (13/50), 28.0% (14/50), and 34.0% (17/50), respectively. Compared with the control group at the same time points, the early recurrence rates of AF in the treatment group were significantly lower at 4 weeks, 8 weeks, and 12 weeks after surgery (P<0.01). Compared with the baseline within group, BNP, hs-CRP, IL-6, and TNF-α in the treatment group all decreased after 12 weeks of treatment (P<0.05); the difference in hs-CRP levels (before vs. after treatment) in the treatment group was higher than that in the control group (P<0.01). Compared with the baseline within group, both groups showed decreases in the total score of AFEQT Questionnaire, scores of the daily life dimension, treatment worry dimension, symptom dimension, and TCM syndrome scores at 4 weeks, 8 weeks, and 12 weeks after treatment. Meanwhile, the score of the treatment satisfaction dimension of AFEQT increased in both groups (P<0.01), and the improvements in all the above scores in the treatment group were superior to those in the control group at all time points (P<0.05 or P<0.01). All safety indicators of patients in both groups were within the normal range before treatment and at 12 weeks after treatment, and no adverse reactions or adverse events occurred in either group. ConclusionModified Zhigancao Granules can reduce the early recurrence rate following radiofrequency ablation in AF patients with qi-yin deficiency syndrome, improve clinical symptoms and quality of life, suppress inflammatory response, and show good safety.
4.Relationship between serum miR-9-5p,miR-21-5p and miR-206 and hemorrhagic transformation occurs and prognosis in patients with large artery atherosclerotic cerebral infarction after thrombolysis
Wenming XU ; Xing WEI ; Dacai GONG ; Bin GE ; Chunling GAO
International Journal of Laboratory Medicine 2025;46(16):1933-1940
Objective To investigate the relationship between serum microRNA(miRNA,miR)-9-5p,miR-21-5p and miR-206 and hemorrhagic transformation(HT)occurs and prognosis in patients with large ar-tery atherosclerotic cerebral infarction(ACI-LAA)after thrombolysis.Methods A total of 265 patients with ACI-LAA admitted to the hospital from April 2021 to November 2023 were selected as the research objects.According to whether HT occurred after intravenous thrombolysis,they were divided into non-HT group and HT group,and the expression levels of serum miR-9-5p,miR-21-5p and miR-206 were compared between the two groups.The general data of patients with ACI-LAA were collected.Univariate and multivariate Logistic regression analysis were used to analyze the influencing factors of HT occurs in patients with ACI-LAA after thrombolysis.The receiver operating characteristic(ROC)curve was used to analyze the efficacy of serum miR-9-5p,miR-21-5p and miR-206 expression levels in predicting HT occurs after thrombolysis in patients with ACI-LAA.The patients with ACI-LAA were followed up for 90 d.According to the modified Rankin scale(mRS)score,the patients were divided into a good prognosis group and a poor prognosis group,and the expression levels of serum miR-9-5p,miR-21-5p and miR-206 were compared between the two groups.Univa-riate and multivariate Logistic regression analysis were used to analyze the influencing factors of poor progno-sis in patients with ACI-LAA.ROC curve was used to analyze the efficacy of serum miR-9-5p,miR-21-5p and miR-206 expression levels in predicting poor prognosis of patients with ACI-LAA.Results HT occurred in 74(27.92%)of 265 patients with ACI-LAA after thrombolysis.Compared with the non-HT group,the National Institutes of Health Stroke Scale(NIHSS)score atadmission,the proportion of hypertension,the proportion of infarct size ≥10 cm2,and the expression levels of serum miR-21-5p and miR-206 were significantly in-creased in the HT group(P<0.05).However,the expression level of serum miR-9-5p was decreased(P<0.05).Hypertension,high NIHSS score at admission,the proportion of infarct size ≥10 cm2,high expression of miR-21-5p and miR-206 were risk factors for HT occurs in patients with ACI-LAA after intravenous thrombolysis,and high expression of miR-9-5p was a protective factor(P<0.05).The ROC curve showed that the area under the curve(AUC)and 95%CI of single and combined detection of serum miR-9-5p,miR-21-5p and miR-206 for predicting HT occurs were 0.796(0.726-0.866),0.779(0.711-0.846),0.784(0.714-0.854),0.891(0.839-0.943),respectively.Among 265 patients with ACI-LAA,83 patients(31.32%)had poor prognosis at 90 d of follow-up.Compared with the good prognosis group,the age,NIHSS score at admis-sion,the proportion of HT,the proportion of infarct size ≥10 cm2,and the expression levels of serum miR-21-5p and miR-206 were increased(P<0.05),and the expression level of serum miR-9-5p was decreased(P<0.05)in the poor prognosis group.Advanced age,high NIHSS score at admission,HT,infarct size ≥10 cm2,high expression of miR-21-5p,and high expression of miR-206 were risk factors for poor prognosis of ACI-LAA patients,and high expression of miR-9-5p was a protective factor(P<0.05).The ROC curve showed that The AUC(95%CI)of single and combined detection of serum miR-9-5p,miR-21-5p and miR-206 for predicting poor prognosis were 0.754(0.691-0.818),0.779(0.719-0.838),0.792(0.815-0.919),0.931(0.902-0.960),respectively.Conclusion Low expression of serum miR-9-5p,high expression of miR-21-5p and high expression of miR-206 are associated with the HT occurs and poor prognosis in patients with ACI-LAA after thrombolysis,and the combined detection of the three indicators has a high predictive value for the HT occurs and poor prognosis.
5.Protocol for clinical practice guideline for the diagnosis and treatment of osteoporotic distal radius fracture through integrated traditional Chinese medicine and Western medicine
Lyuyu LI ; Chuan LI ; Dan XING ; Bin WANG ; Ning LI ; Yan YAN ; Weiheng CHEN ; Yongqing XU ; Wei DONG
Chinese Journal of Trauma 2025;41(3):253-258
Osteoporotic distal radius fracture (ODRF), a common fragility fracture in elderly patients, poses a significant threat to patients′ life and health due to its high incidence and disability rate. Both traditional Chinese medicine and Western medicine have accumulated extensive clinical experience in the diagnosis, treatment and rehabilitation of ODRF. However, there is currently a lack of specific guideline on how to effectively integrate the strengths of the two medical systems. To standardize the collaborative diagnosis and treatment of ODRF using integrated traditional Chinese and Western medicine, the Clinical Practice Guideline Working Group for the clinical practice guideline for the diagnosis and treatment of osteoporotic distal radius fracture through integrated traditional Chinese medicine and Western medicine was established in September 2024, led by the Third Affiliated Hospital of Yunnan University of Chinese medicine (Kunming Hospital of Chinese Medicine) with the participation of many medical institutions across China. Protocol for the guideline was then formulated to detail each key aspect in the development of the guideline and explain the related procedures, aiming to ensure a standardized, orderly and transparent development.
6.Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique (version 2025)
Sihao HE ; Junchao XING ; Tongwei CHU ; Zhengqi CHANG ; Xigao CHENG ; Fei DAI ; Xiaobing JIANG ; Jie HAO ; Jiang HU ; Jinghui HUANG ; Tianyong HOU ; Fei LUO ; Bo LIAO ; Changqing LI ; Lei LIU ; Guodong LIU ; Peng LIU ; Sheng LU ; Weishi LI ; Yang LIU ; Zhen LIU ; Wei MEI ; Peifu TANG ; Bing WANG ; Bing WANG ; Ce WANG ; Hongli WANG ; Liang WANG ; Shengru WANG ; Xiaobin WANG ; Yang WANG ; Yingfeng WANG ; Zheng WANG ; Jianzhong XU ; Guoyong YIN ; Haiyang YU ; Qiang YANG ; Zhaoming YE ; Bin ZHANG ; Chengmin ZHANG ; Jun ZOU ; Qiang ZHOU ; Min ZHAO ; Rui ZHOU ; Xiaojun ZHANG ; Yongfei ZHAO ; Zhongrong ZHANG ; Zehua ZHANG ; Yingze ZHANG
Chinese Journal of Trauma 2025;41(11):1035-1047
For middle-aged and elderly patients with conditions such as spinal fractures and degenerative spinal diseases, spinal internal fixation is a core surgical procedure for reconstructing spinal stability, heavily relying on the biomechanical stability provided by pedicle screw systems. Whereas, these patients are often complicated by osteoporosis that can significantly compromise the stability of the bone-pedicle screw interface, leading to a marked increase in pedicle screw loosening and surgical failure rates. The bone cement-augmented pedicle screw technique, which involves injecting bone cement into the vertebral body or screw trajectory to optimize the mechanical properties of the bone-pedicle screw composite, has been proven to significantly enhance fixation strength and effectively prevent screw-related failures, thereby reducing the incidence of internal fixation failure in high-risk populations undergoing spinal fusion. However, the widespread clinical application of this technique has faced challenges such as inaccurate clinical decision-making (indication and contraindication selection), non-standardized operative practices, and insufficient awareness of complication prevention, resulting in considerable variability in clinical outcomes and even severe complications. To address this, Prof. Luo Fei from First Affiliated Hospital of Army Medical University initiated the project and the Chinese Association Orthopaedic Surgeons organized relevant experts to develop the Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique ( version 2025), based on current evidence. The guidelines put forward 8 recommendations regarding the clinical value, scope of application, and operational standards of the technique, aiming to provide evidence-based medical support and technical standardization for clinical decision-making.
7.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
8.Robot-assisted surgery for renal cell carcinoma in a pilot: a case report and literature review
Xintao LI ; Jinxuan ZHANG ; Haibo SHENG ; Jun WANG ; Di LI ; Jizhang XING ; Jingmin YAN ; Dawei MU ; Kaikai CHEN ; Xiaolong WEI ; Shuwei XIAO ; Bin SUN ; Jianye LI
Chinese Journal of Aerospace Medicine 2025;36(2):145-149
Objective:To investigate the selection of treatment methods for renal tumors in pilots as well as the clinical significance of robot-assisted surgery by summarizing the process of robot-assisted surgery in the treatment of renal cell carcinoma in a pilot.Methods:The diagnosis, robot-assisted surgery and aeromedical assessment of a pilot with renal cell carcinoma were reported, and the related literature was reviewed.Results:The patient was a 44-year-old male transporter pilot, who was diagnosed with a left renal mass in the middle-lower pole of the kidney during a routine abdominal CT scan. After detailed preoperative evaluation that ruled out the possibility of distant metastasis and other surgical contraindications, the patient underwent robot-assisted laparoscopic partial nephrectomy in August 2022. The postoperative recovery went well, and renal function remained within normal limits at follow-ups. In March 2023, the pilot was concluded as qualified for flight after aeromedical assessment.Conclusions:Robot-assisted partial nephrectomy can significantly reduce surgical trauma, lower the risk of complications, and maximally preserve renal function. It is a good approach to renal tumors in pilots who can recover quickly.
9.A clinical study of deep learning image reconstruction algorithms in liver dual-energy CT with reduced radiation dose to further improve image quality and lesion diagnostic confidence
Yuncheng LI ; Yuguo LI ; Junlin YANG ; Jian SONG ; Xing TANG ; Wei DENG ; Zhen WANG ; Jinxiu YANG ; Bin LIU ; Yongqiang YU ; Xiaohu LI
Chinese Journal of Radiology 2025;59(1):43-49
Objective:To explore the feasibility of applying deep learning image reconstruction (DLIR) in low-radiation dose liver dual-energy CT to further improve image quality, diagnostic confidence of lesion, and accuracy of iodine concentration (IC) measurement.Methods:This prospective cohort study enrolled 60 patients scheduled for enhanced liver CT at the First Affiliated Hospital of Anhui Medical University from June 2023 to January 2024. The participants were randomly assigned into the standard dose group and low radiation dose group with 30 cases in each using randomized block method. The standard radiation dose group underwent standard-radiation dose 120 kVp scans during the venous phase, while the low radiation dose group underwent low radiation dose scans with a rapid kVp-switching spectral scanning mode at 80 kVp and 140 kVp. The effective radiation dose (ED) was calculated for both groups. The standard radiation dose group was reconstructed using adaptive statistical iterative reconstruction-V (ASIR-V) algorithm 40% (AR40 120 kVp). The low radiation dose group using high-intensity DLIR (DLIR-H) to reconstructed 40 keV and 50 keV virtual monoenergetic images (VMI) (DH-VMI 40 keV, DH-VMI 50 keV). The image quality of the above three groups was objectively evaluated through the measurement of image noise and calculation of contrast-to-noise ratio (CNR) and signal-to-noise ratio (SNR) for the liver and portal vein; and the image quality was subjectively scored for image noise, contrast, lesion conspicuity, and diagnostic confidence. In the low radiation dose group, DLIR-H and ASIR-V40% reconstructed iodine maps were used to measure the liver and portal vein of IC values, standard deviations (SD), and coefficients of variation (CV). One-way analysis of variance or Kruskal-Wallis H test was used to compare the differences of subjective and objective image quality among the three groups, and paired t-test was used to compare the differences in measurement indexes between DLIR-H and ASIR-V40% reconstructed iodine maps. Results:The ED in the low radiation dose group [(2.2±0.5) mSv] was reduced by 56.8% compared to the conventional radiation dose group [(5.4±1.4) mSv]. Objective evaluations demonstrated that DH-VMI 40 keV had higher image noise, CNR, and SNR for liver and portal veins compared to AR40 120 kVp ( P<0.001). DH-VMI 50 keV had lower image noise ( P=0.200), with higher CNR and SNR for the liver and portal vein compared to AR40 120 kVp( P<0.001). In subjective evaluation, there was no statistically significant difference in image noise scores between DH-VMI 40 keV and AR40 120 kVp ( P>0.05), while the image noise score for DH-VMI 50 keV was lower than that of AR40 120 kVp ( P<0.05). Both DH-VMI 40 keV and DH-VMI 50 keV had higher scores for contrast, lesion conspicuity, and diagnostic confidence compared to those of AR40 120 kVp ( P<0.05). In the low radiation dose group, there was no statistically significant difference in IC values for the liver and portal vein between the ASIR-V40% and DLIR-H algorithm reconstructed iodine maps ( P>0.05). The SD and CV of liver and portal vein in the DLIR-H reconstructed iodine maps were lower than those in the ASIR-V40% reconstructed iodine maps ( P<0.001). Conclusions:DLIR can effectively reduce the image noise of low-energy (40, 50 keV) VMI, enhance lesion conspicuity and diagnostic confidence, and improve measurement accuracy without affecting IC values.
10.Construction of p97 mutant of Mesomycoplasma hyopneumoniae based on the homologous recombination system
Yanna WEI ; Jiying WANG ; Huan XIE ; Zhiqiang LI ; Z.A.Ishag HASSAN ; Xing XIE ; Bin XU ; Qiyan XIONG ; Zhixin FENG ; Guoqing SHAO ; Yanfei YU
Chinese Journal of Veterinary Science 2025;45(3):473-481
The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

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