1.High-throughput Sequencing of MET Mutational Landscape in Cancer Patients and the Value of IHC Screening for Targeted Therapy
Feicheng YANG ; Lixia JIANG ; Ruobing LIU ; Kai YANG ; Yewei LI ; Yi XIONG ; Hao DENG ; Jie HU
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(1):172-181
ObjectiveTo investigate the mutation status of the MET gene in tumor patients, with a focus on its expression in lung cancer tissues and its impact on patient prognosis, providing theoretical basis and potential targets for precise diagnosis and treatment of the tumors. MethodsA total of 1,627 tumor patient samples were collected from January 2021 to September 2025. High-throughput sequencing technology was used to conduct DNA-level gene detection on 137 tumor patients with MET mutations. The distribution of mutation sites in exons and introns, as well as the occurrence frequencies of MET gene amplification and gene fusion were analyzed, with particular attention paid to the mutation frequencies of each exon and the high-frequency variant types. Immunohistochemical staining was performed to analyze the expression levels of c-MET protein in tumor tissues of the 137 patients. Immunohistochemical analysis was conducted on tumor tissues of 9 patients with MET gene exon 14 skipping mutations to study the association between c-MET protein expression and this specific mutation type. RNA sequencing results, immunohistochemical staining data, and follow-up prognosis of lung cancer from the GEPIA2 database and TCGA database were comprehensively analyzed to explore the potential relationship between MET gene expression in tumor tissues and patient prognosis. ResultsAmong 1,627 tumor patients, 137 MET mutation patients were detected, including 93 males with an average age of (66.85±9.61) years and 44 females with an average age of (59.43±11.08) years. Lung cancer was the most common tumor type (117 cases). MET gene expression was lower in breast cancer and glioblastoma multiforme, but higher in lung cancer and colorectal cancer as compared with the expression in normal tissues. Survival analysis showed that patients with high MET gene expression in lung adenocarcinoma had a shorter overall survival time than those with low expression. A total of 961 mutations were detected by next-generation sequencing, including 547 exon mutations and 374 intron mutations. MET gene amplification was found in 13 cases and gene fusion in 27 cases. The highest mutation frequency was in exon 2, followed by exon 5, exon 4, and exon 19. High-frequency variant types included c.2890C>A (p.L964M) on exon 4 and c.3028G>T (p.D1010Y) on exon 5. The c-MET protein expression was closely related to the type of gene mutation. Missense mutations often led to positive or strongly positive c-MET protein expression, while nonsense mutations often resulted in weak or absent c-MET protein expression. Among the 9 patients with MET gene exon 14 skipping mutations, the c-MET protein expression score was mostly 0 - 1, with only 1 case scoring 2. Among the 117 lung cancer patients, the incidence rate was higher in males than in females, and more common among stage Ⅲ-Ⅳ patients than among stage I-Ⅱ patients. Out of the 41 patients treated with surgery + targeted drugs (gutemitinib, savolitinib), 8 cases of recurrence or metastasis were found during follow-up. ConclusionMET gene mutations exhibit specific mutation characteristics in different tumor types, and its expression levels show significant differences between various tumor tissues and normal tissues. It is closely related to the poor prognosis of lung adenocarcinoma patients. We found that c-MET protein expression is closely related to the type of MET gene mutation, especially in patients with MET gene exon 14 skipping mutations, which presents a characteristic expression pattern. This provides an important basis for the preliminary screening using immunohistochemistry in clinical practice.
2.Overview of Real-time Delphi Method and Its Application in Guidelines
Haiyun WANG ; Ruobing LEI ; Xuan YU ; Hui LIU ; Qiu LI ; Yaolong CHEN
Medical Journal of Peking Union Medical College Hospital 2025;16(6):1571-1577
The real-time Delphi method represents a refinement of the classical Delphi technique, designed to overcome limitations such as prolonged study duration and delayed feedback during consensus development. This article, building upon the classical Delphi foundation, systematically elaborates on the application process, advantages, and limitations of the real-time Delphi method. It further presents currently available websites or software capable of facilitating real-time Delphi exercises and offers considerations and recommendations for its application in guideline development, aiming to serve as a reference for relevant researchers.
3.Role and mechanism of dexmedetomidine in regulating bone metabolism in tail-suspended osteoporotic rats
Yunliang GUO ; Can WANG ; Xinyu ZHANG ; Zedong YAN ; Zhipeng WEN ; Ruobing LIU ; Pengsen LIU
Journal of Army Medical University 2025;47(3):226-233
Objective To investigate the effect of dexmedetomidine(Dex)on bone loss in tail-suspended rats and primarily explore its regulatory mechanism on bone metabolism.Methods A total of 30 male rats were randomly divided into a control group,a model group,and a Dex group,with 10 animals in each group.Rat model of osteoporosis was established by hind limb suspension for 4 weeks.Dex at a dose of 10 μg/kg was given intraperitoneally,once every other day from the day of tail suspension.And equal amount of normal saline was given to the control and model group.Bone histological staining was used to observe the trabecular bone area fraction.Biomechanical three-point bending test was employed to measure the maximum load,stiffness,and fracture energy.Dual calcein/alizarin red fluorescence labeling and tartrate resistant acid phosphatase(TRAP)staining were applied respectively to detect the mineral apposition rate and bone formation rate as well as the number of osteoclasts on bone surfaces.Secondly,after primary osteoblasts were isolated from the tibiae of tail-suspended rats and then treated with 1 nmol/L Dex,the proportion of alkaline phosphatase(ALP)-positive osteoblasts and the activity of the enzyme were detected by ALP staining and activity test.qRT-PCR was applied to measure the expression of osteogenic activity-related factors,including osteocalcin(Ocn),Runt related transcription factor 2(Runx2),Osterix protein(Osx),and type 1 collagen(Col1).Results The animal experiments revealed that Dex treatment significantly increased the tibial trabecular bone area fraction,inhibited the decrease in bone mechanical strength,and enhanced the mineralization deposition rate and new bone formation rate of trabecular bone in the tail-suspended rats(all P<0.001).The in vitro experiments showed that Dex treatment obviously improved ALP activity and the number of ALP-positive osteoblasts in primary osteoblasts isolated from tail-suspended rats(P<0.01),and up-regulated the expression levels of osteogenic differentiation-related genes,such as Ocn,Runx2,Osx and Col1(P<0.01).Conclusion Dex exerts anti-bone loss effect in tail-suspended rats,which may be associated with its stimulation on osteoblast-mediated bone formation.
4.Long-term effect of modified Morrow surgery on hypertrophic obstructive cardiomyopathy in children: A retrospective study in a single center
Xiaoyi LI ; Hongxiang WU ; Ruobing WANG ; Haozhong LIU ; Xiaodong ZENG ; Ying ZENG ; Shengwen WANG ; Minjie HUANG ; Haiyun YUAN ; Jian LIU ; Huiming GUO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(07):985-990
Objective To analyze the long-term outcome of modified Morrow surgery (interventricular septal cardiomyectomy) in the treatment of hypertrophic obstructive cardiomyopathy (HOCM) in children. Methods The clinical data of the children with HOCM (aged≤14 years) who underwent modified Morrow surgery from January 2010 to August 2022 in Guangdong Provincial People's Hospital were retrospectively analyzed, including changes in hospitalization status, perioperative period, and long-term 15-lead electrocardiogram and echocardiography. Results A total of 29 patients were collected, including 22 males and 7 females, aged 10.00 (5.00, 12.00) years. Five (17.9%) patients had New York Heart Association (NYHA) heart function grade Ⅲ or Ⅳ. Ventricular septal cardiomyectomy was performed in all patients. All 29 patients survived and their cardiac function recovered after operation. Before discharge, right bundle branch block was observed in 2 patients and left bundle branch block in 6 patients. After surgery, in the left ventricular septal cardiomyectomy, the left atrial diameter decreased (P<0.001), left ventricular end-systolic diameter increased (P=0.009), the peak pressure gradient of left ventricular outflow tract decreased (P<0.001), and the thickness of ventricular septum decreased (P<0.001). The systolic anterior motion of mitral valve disappeared and mitral regurgitent jet area decreased (P<0.001). The flow velocity and peak pressure gradient of right ventricular outflow tract also decreased in the patients who underwent right ventricular septal cardiomyectomy. The average follow-up of the patients was 69.03±10.60 months. All the patients survived with their NYHA cardiac function grading Ⅰ or Ⅱ. No new-onset arrythmia event was found. Echocardiography indicated that the peak pressure gradient of the left ventricular outflow tract remained low (P<0.001). Moderate mitral regurgitation occurred in 2 patients, and left ventricular outflow tract obstruction with moderate mitral regurgitation occurred in 1 patient after simple right ventricular septal cardiomyectomy. Conclusion Right ventricular or biventricular obstruction is frequent in the children with HOCM and they usually have more symptoms before surgery. Modified Morrow surgery can effectively relieve outflow tract obstruction and improve their cardiac function. The long-term outcome is satisfactory. However, the posterior wall of the left ventricle remains hypertrophic. Also, there is an increased risk of a conduction block.
5.Progress of pancreatic surgical treatment in 2023
Ruobing WANG ; Yueze LIU ; Taiping ZHANG
Chinese Journal of General Surgery 2024;39(1):14-18
Pancreatic surgery is one of the most challenging specialties in general surgery. Due to the variety of pancreatic diseases, the difficulty of surgery, and the differences in diagnosis and treatment among different diseases, treatment strategies for these diseases remain controversial. From the aspects of surgical treatment such as pancreatitis, pancreatic cystic neoplasms, pancreatic neuroendocrine neoplasms, and pancreatic cancer, as well as neoadjuvant therapy, adjuvant therapy, immunotherapy, and targeted therapy. We review and summarize the frontier progress of clinical and translational research in pancreatic surgery in 2023, in order to further standardize the diagnosis and treatment of pancreatic surgery.
6.Clinical effects of anterolateral femoral or anteromedial femoral perforator flaps in repairing skin and soft tissue defects after resection of cutaneous squamous cell carcinoma
Ruobing LIU ; Deyi ZHENG ; Baoyun WANG ; Weiren LI
Chinese Journal of Burns 2024;40(6):564-571
Objective:To explore the clinical effects of anterolateral femoral or anteromedial femoral perforator flaps in repairing skin and soft tissue defects after resection of cutaneous squamous cell carcinoma (CSCC).Methods:This study was a retrospective observational study. From July 2015 to July 2022, 21 patients with CSCC were treated in the Department of Burns and Plastic Surgery of Guizhou Provincial People's Hospital, including 15 males and 6 females, aged from 27 to 74 years. The area of skin and soft tissue defects after extended resection of CSCC was 7.5 cm×4.0 cm to 23.0 cm×8.5 cm. The wounds in 18 patients were repaired with anterolateral femoral perforator flaps; variations of perforating branch of the descending branch of lateral circumflex femoral artery were observed in 3 patients during the operation, and the wounds were repaired with anteromedial femoral perforator flaps. The flap areas were 8.0 cm×5.0 cm to 25.0 cm×10.0 cm. The wounds in the donor areas were sutured directly in 19 patients, and the wounds in the donor areas were repaired with thin and medium-thickness skin grafts in the contralateral thigh in 2 patients. The postoperative survival of flaps and the occurrence of vascular crisis were observed. The length of operation and the hospitalization day were recorded. The recurrence of tumor, the appearances of the donor and recipient areas of flaps, the function of the flap donor area were followed up. At the last follow-up, the satisfaction degree of patients for the curative effects was evaluated.Results:The flaps survived in 20 patients, while the vascular crisis occurred in 1 patient within 48 hours after operation, and the flap survived after immediate emergency operation. The length of operation was 4 to 5 hours, and the hospitalization day was 15 to 38 days. The patients were followed up for 1 to 6 years after operation, there was no local tumor recurrence, the color and texture of the flaps were with no obvious differences to those of the surrounding tissue, and the elasticity and appearance were good. The skin grafts in the flap donor areas of 2 patients survived well with local pigmentation. There was only linear scar in the flap donor areas of all patients, and there were no significant effects on sensory and motor functions. At the last follow-up, fifteen patients were satisfied with the curative effect, and 6 patients were generally satisfied with the curative effect.Conclusions:For skin and soft tissue defects after CSCC resection, the anterolateral femoral perforator flaps can be used preferentially. In the case of variation of the perforating branch of descending branch of the lateral circumflex femoral artery, the anteromedial femoral perforator flap is selected. The areas of the two flaps are large and can be adjusted according to the amount of defect tissue, thus accurately and effectively repairing skin and soft tissue defects after CSCC resection. The postoperative appearance and function are good.
7.Role of connexin 43 in a rat model of periodontitis-induced renal injury
Yu XIN ; Ruobing FU ; Xirui XIN ; Yaqi SHANG ; Xinchan LIU ; Weixian YU
West China Journal of Stomatology 2024;42(3):296-303
Objective This study aims to investigate the role of gap junction mediated by connexin 43(Cx43)in renal injury induced by periodontitis in rats.Methods Twelve SPF-grade Wistar male rats were divided into a control group and a periodontitis group by using a completely random number table method,with six rats in each group.The control group rats were not treated,while the periodontitis group rats were subjected to wire ligation of the neck of their bilateral maxillary first molars to construct a periodontitis model.After 8 weeks of modeling,the rats were examined for clinical indicators of the periodontium.micro-CT scanning of the maxilla reconstructed its 3D structure and analyzed the absorption of alveolar bone.Histopathological changes in periodontal and renal tissues were detected.MitoSOX red reagent was used to determine reactive oxygen species(ROS)content in renal tissues.A biochemical reagent kit was used to detect serum oxidative stress biomarkers.Real-time fluo-rescent quantitative-polymerase chain reaction(qRT-PCR)was employed to determine Cx43,nuclear factor kappa-B(NF-κB),interleukin(IL)-1β,IL-6,BCL2-Associated X(Bax),B-lymphomatoma-2 gene(Bcl-2),and Caspase-3 mRNA were determined.Western blot analysis was used to detect Cx43,NF-κB,IL-1β,Bax,Bcl-2 and Caspase-3 protein.Re-sults micro-CT 3D reconstruction showed significant bone resorption of the first molar alveolar bone in the periodonti-tis group rats and decreased height of the alveolar ridge.The distance from the enamel cementum boundary to the top of the alveolar ridge in the periodontitis group was significantly higher than that inthe control group.The histopathological results showed a large number of inflammatory cells that infiltrated the periodontal tissue of the periodontitis group,and the alveolar bone was significantly absorbed.Rats in the periodontitis group also exhibited mild thickening of the glomer-ular basement membrane,dilation of the Bowman's capsule,and destruction of the brush-like edge of the renal tubules in the renal tissue.The MitoSOX red staining results showed a significant increase in ROS content in the renal tissue of the periodontitis group.The biochemical test results showed that the levels of superoxide dismutase and glutathione in the serum of rats with periodontitis decreased,while that of malondialdehyde increased.The results of qRT-PCR and Western blot showed that the expression levels of Cx43,IL-1β,IL-6,Bax,Caspase-3 mRNA and Cx43,IL-1β,NF-κB,Bax,Caspase-3 proteins in the periodontitis group significantly increased compared with those in the control group,while the expression levels of Bcl-2 mRNA and protein decreased.Conclusion Periodontitis may activate NF-κB sig-naling molecules by upregulating the expression of Cx43 in rat kidney tissues,leading to increased levels of inflamma-tion and apoptosis and ultimately inducing kidney injury.
8.A Protocol for Developing Chinese Clinical Practice Guidelines of Hypertension
Ying LOU ; Wenjun MA ; Zijun WANG ; Nan YANG ; Yajia SUN ; Yunlan LIU ; Ruobing LEI ; Junxian ZHAO ; Xufei LUO ; Lu WANG ; Yaolong CHEN ; Yaling HAN ; Yingxian SUN ; Yuming LI ; Jun CAI
Cardiology Discovery 2024;04(3):187-191
To improve the standard screening, diagnosis, and treatment of hypertension in patients in China; realize the standardization of clinical practice of hypertension; and improve the prevention and control level of hypertension in China, it is both important and necessary to develop a clinical practice guideline for hypertension according to a recognized methodology. Jointly sponsored by the National Center for Cardiovascular Diseases, Chinese Medical Doctor Association, Hypertension Committee of the Chinese Medical Doctor Association, Chinese Society of Cardiology, and Hypertension Committee of Cross-Straits Medicine Exchange Association, the "Chinese Clinical Practice Guidelines of Hypertension" was proposed. Research Unit of Evidence-Based Evaluation and Guidelines, Chinese Academy of Medical Sciences, Guideline and Standards Research Centre of Chinese Medical Association Publishing House, Lanzhou University Institute of Health Data Science, and Lanzhou University GRADE Center will provide methodological support for the guidelines.
9.A Protocol for Developing Chinese Clinical Practice Guidelines of Hypertension
Ying LOU ; Wenjun MA ; Zijun WANG ; Nan YANG ; Yajia SUN ; Yunlan LIU ; Ruobing LEI ; Junxian ZHAO ; Xufei LUO ; Lu WANG ; Yaolong CHEN ; Yaling HAN ; Yingxian SUN ; Yuming LI ; Jun CAI
Cardiology Discovery 2024;04(3):187-191
To improve the standard screening, diagnosis, and treatment of hypertension in patients in China; realize the standardization of clinical practice of hypertension; and improve the prevention and control level of hypertension in China, it is both important and necessary to develop a clinical practice guideline for hypertension according to a recognized methodology. Jointly sponsored by the National Center for Cardiovascular Diseases, Chinese Medical Doctor Association, Hypertension Committee of the Chinese Medical Doctor Association, Chinese Society of Cardiology, and Hypertension Committee of Cross-Straits Medicine Exchange Association, the "Chinese Clinical Practice Guidelines of Hypertension" was proposed. Research Unit of Evidence-Based Evaluation and Guidelines, Chinese Academy of Medical Sciences, Guideline and Standards Research Centre of Chinese Medical Association Publishing House, Lanzhou University Institute of Health Data Science, and Lanzhou University GRADE Center will provide methodological support for the guidelines.
10.Crohn′s disease with NLRP12 genetic variation: report of 2 cases with literature review
Ruobing LIU ; Xiangsu LI ; Yang HUANG ; Ailan LI ; Fen WEN ; Xue ZHAO ; Chaonan WANG ; Xueyi XIAO ; Qingqing YANG ; Xudong WU
Chinese Journal of Inflammatory Bowel Diseases 2024;08(1):101-104
A pair of father-son Crohn′s disease (CD) patients with NOD-like receptors family pyrin domain containing 12 ( NLRP12) gene c.1382 mutation was reported. Through the relevant literature review, we summarize the other CD patients complicated with NLRP12 genetic variation at home and abroad and the mechanism of NLRP12 in inflammatory bowel disease. This study aims to provide reference for the subsequent exploration of individulized treatment.

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