1.Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA
Jiayan XIONG ; Wei LEI ; Bo YOU ; Zhenxin ZHANG ; Haijing XIE ; Ying SHAN ; Tian XIA ; Yong ZHOU
China Oncology 2024;34(5):451-459
Background and purpose:DDX is an adenosine triphosphate(ATP)-dependent RNA helicase closely related to mRNA regulation,tumor proliferation and invasion.This article aimed to explore the effect of DDX6,a member of the DDX family,on the stability of CKMT1A mRNA,as well as the effect of the DDX6 CKMT1A axis on the proliferation and migration ability of human nasopharyngeal carcinoma cell CNE2 and its molecular mechanism.Methods:We retrieved the data of expressions of DDX6 and CKMT1A in human head and neck squamous cell carcinoma from The Cancer Genome Atlas(TCGA)database and performed a correlation analysis.Western blot was performed to detect the expressions of CKMT1A and DDX6 in human nasopharyngeal carcinoma tissues and normal nasopharyngeal tissues preserved by Affiliated Hospital of Nantong University.This study was approved by the Ethics Committee of Affiliated Hospital of Nantong University(Number:2022-L114).We used transwell assay to detect cell migration ability,EdU assay to detect cell proliferation ability,and colony formation assay to detect clone formation ability.We transfect with lentivirus and plasmids to construct sh-DDX6,sh-CKMT1A,sh-CKMT1A+sh-DDX6 and oe-CKMT1A cell models derived from the human nasopharyngeal carcinoma cell line CNE2,preserved by Affiliated Hospital of Nantong University,to clarify the impact of DDX6 and CKMT1A expression levels on the malignant biological phenotypes of nasopharyngeal carcinoma cells.BALB/c nude mice subcutaneous xenograft tumor model was constructed to detect the effects of DDX6 and CKMT1A on nasopharyngeal carcinoma cells in mice.RNA stability assay was used to detect the effect of DDX6 knockout on CKMT1A mRNA and further clarify the molecular mechanism of DDX6.Results:DDX6 was highly expressed,CKMT1A level was low in human nasopharyngeal carcinoma tissue,and DDX6 was negatively correlated with CKMT1A expression.DDX6 inhibited protein translation of CKMT1A by disrupting its mRNA stability.Low expression of CKMT1A in CNE2 cells enhanced cell migration and proliferation ability,while high expression inhibited migration and proliferation ability.Knocking out DDX6 reversed the progression of malignant behavior caused by downregulation of CKMT1A.Low expression of CKMT1A promoted tumor cell growth in BALB/c nude mice subcutaneous xenograft tumor model,while low expression of DDX6 inhibited tumor cell growth.Knocking out DDX6 and CKMT1A simultaneously restored the inhibitory effect caused by knocking down DDX6 alone.Conclusion:DDX6 in nasopharyngeal carcinoma cells disrupts the stability of CKMT1A mRNA,negatively regulates CKMT1A protein translation,upregulates the proliferation and migration ability of nasopharyngeal carcinoma cells,and promotes malignant progression of nasopharyngeal carcinoma.
2.Hepatocellular carcinoma prediction model performance decreases with long-term antiviral therapy in chronic hepatitis B patients
Xiaoning WU ; Xiaoqian XU ; Jialing ZHOU ; YaMeng SUN ; Huiguo DING ; Wen XIE ; Guofeng CHEN ; Anlin MA ; HongXin PIAO ; Bingqiong WANG ; Shuyan CHEN ; Tongtong MENG ; Xiaojuan OU ; Hwai-I YANG ; Jidong JIA ; Yuanyuan KONG ; Hong YOU
Clinical and Molecular Hepatology 2023;29(3):747-762
Background/Aims:
Existing hepatocellular carcinoma (HCC) prediction models are derived mainly from pretreatment or early on-treatment parameters. We reassessed the dynamic changes in the performance of 17 HCC models in patients with chronic hepatitis B (CHB) during long-term antiviral therapy (AVT).
Methods:
Among 987 CHB patients administered long-term entecavir therapy, 660 patients had 8 years of follow-up data. Model scores were calculated using on-treatment values at 2.5, 3, 3.5, 4, 4.5, and 5 years of AVT to predict threeyear HCC occurrence. Model performance was assessed with the area under the receiver operating curve (AUROC). The original model cutoffs to distinguish different levels of HCC risk were evaluated by the log-rank test.
Results:
The AUROCs of the 17 HCC models varied from 0.51 to 0.78 when using on-treatment scores from years 2.5 to 5. Models with a cirrhosis variable showed numerically higher AUROCs (pooled at 0.65–0.73 for treated, untreated, or mixed treatment models) than models without (treated or mixed models: 0.61–0.68; untreated models: 0.51–0.59). Stratification into low, intermediate, and high-risk levels using the original cutoff values could no longer reflect the true HCC incidence using scores after 3.5 years of AVT for models without cirrhosis and after 4 years of AVT for models with cirrhosis.
Conclusions
The performance of existing HCC prediction models, especially models without the cirrhosis variable, decreased in CHB patients on long-term AVT. The optimization of existing models or the development of novel models for better HCC prediction during long-term AVT is warranted.
3.Clinical application of 99Tc m-MIBI SPECT/CT muscle imaging in the diagnosis of dystonic muscle of spasmodic torticollis
Quanpeng WANG ; Bing ZHANG ; Tingting SHEN ; Xuan ZHOU ; Yichi XIE ; Jiaxi YOU ; Zengli LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2023;43(10):593-598
Objective:To explore the potential clinical value of 99Tc m-methoxyisobutylisonitrile(MIBI) SPECT/CT muscle imaging in the diagnosis of cervical dystonia (CD). Methods:From January 2021 to April 2022, 50 patients with CD (14 males, 36 females; age (45.8±12.5) years) who were treated in Second Affiliated Hospital of Soochow University were prospectively included. The 99Tc m-MIBI SPECT/CT muscle imaging results of 400 pieces of muscle (bilateral sternocleidomastoid, musculus scapulae, splenius capitis and musculus trapezius; each of 100 pieces) in 50 patients were analyzed and divided into the dystonic muscle group and normal muscle group according to the electromyography (EMG). Toronto western spasmodic torticollis rating scale (TWSTRS) score, SUV max and target-to-background ratio (TBR) of single superficial cervical muscle and total cervical muscle, and EMG diagnosis results were obtained before botulinum toxin injection. ROC curves of SUV max and TBR of dystonic muscles were constructed to determine AUCs and the difference was compared by Delong test. Differences of SUV max and TBR between 2 groups were analyzed by Mann-Whitney U test. Spearman rank correlation analysis was used to analyze the correlation of SUV max, TBR and TWSTRS scores of total cervical muscle. Results:There were 205 pieces of muscle in dystonic muscle group and 195 pieces of muscle in normal muscle group. The uptake of 99Tc m-MIBI in dystonic muscle was significantly increased in CD patients, and the non-whole uptake of 99Tc m-MIBI was increased in some dystonic muscles, which was manifested as uneven uptake of the whole muscle. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of visual analysis were 95.12%(195/205), 75.90%(148/195), 85.75%(343/400), 85.58%(195/242) and 93.67%(148/158), respectively. There were significant differences of SUV max (1.74(1.42, 2.12) vs 0.92(0.81, 0.99)) and TBR (2.55(1.92, 3.27) vs 1.44(1.22, 1.73)) between the dystonic muscle group and the normal muscle group ( z value: -15.29, -12.69, both P<0.001). The diagnostic efficiency of SUV max in dystonic muscle was better than TBR (AUC: 0.942 vs 0.867; z=5.03, P<0.001). SUV max, TBR and TWSTRS score in the neck muscles of patients with CD showed positive correlation ( rs values: 0.44, 0.45, both P<0.001). Conclusion:99Tc m -MIBI SPECT/CT muscle imaging is a good diagnostic method for dystonic muscle in patients with CD.
4.Indications of invasive prenatal diagnosis in third trimester and pregnancy outcome
Yiyun XU ; Xiaoxiao XIE ; Honghui ZHOU ; Longxia WANG ; Yanqin YOU ; Qingdong ZHAO ; Jing SUN ; Yanping LU
Chinese Journal of Perinatal Medicine 2022;25(2):110-116
Objective:To analyze the indications for invasive prenatal diagnosis in the third trimester and summarize the pregnant outcome.Methods:Clinical data of 121 women who underwent invasive prenatal diagnosis in the third trimester in the prenatal diagnostic center of the First Medical Center of Chinese PLA General Hospital from January 2016 to December 2020 was retrospectively analyzed. Different genetic diagnostic methods were used according to different indications. Indications and results of prenatal diagnosis, as well as the complications within two weeks after the invasive procedure, pregnancy outcome, and neonatal follow-up of all the participants were described.Results:Among the 121 cases, 107 cases underwent amniocentesis, seven underwent percutaneous umbilical blood sampling, and seven had both procedures performed at the same time (one underwent thoracocentesis at the same time). Newly identified ultrasound abnormalities in the second and third trimesters were the main indications for prenatal diagnosis, accounting for 99.2%(120/121), of which short limbs and fetal growth restriction accounted for 25.0% (30/120) and 20.0% (24/120), respectively. Genetic abnormalities and congenital diseases were detected in 20 cases with a detection rate of 16.5%(20/121). Among them, there were nine cases of achondroplasia, five cases of pathogenic copy number variations, one case of achondroplasia with pathogenic copy number variation, one trisomy 18, one 47,XXX, one tetrasome mosaicism of 12p, one de novo WTX c. 1072(Exon2) C>Tp.R358X heterozygous mutation, and one fetal hypoproteinemia. In addition, six cases with copy number variation of unknown significance (VUS) were detected, noting for a detection rate of 5.0%(6/121). Among the 20 cases with abnormal detection, 15 were terminated, two delivered prematurely before obtaining the prenatal diagnosis results, one underwent cesarean section before obtaining prenatal diagnostic results and two continued the pregnancies. In the six cases with VUS, one was terminated and the other five continued the pregnancy. Only one case had preterm premature rupture of membranes 2 d after amniocentesis and the incidence rate of complications after all kinds of invasive procedures was 0.8% (1/121). During the neonatal follow-up, postnatal whole exome sequencing revealed monogenetic disorder in two cases with normal prenatal diagnostic results; the patient with 12p chimerism had developmental delay; the one with WTX mutation deceased on the day of born; the rest newborns developed normally. Conclusions:As a relatively safe method, invasive prenatal diagnosis in the third trimester is of great importance and value in reducing the miss diagnostic rate of fetuses with severe genetic diseases and birth defects. The appropriate application of prenatal whole exome sequencing could further help to decrease the miss diagnostic rate of monogenetic disorder.
5. Morphology and tissue structure of acromioclavicular joint
Ri ZHOU ; Xiao-Xiao XIE ; Shuang WANG ; Shi-You REN ; Wen-Tao ZHANG
Acta Anatomica Sinica 2022;53(1):103-107
Objective To explore the morphology and the tissue structure of acromioclavicular joint. Methods Anatomical analysis was performed on 27 fresh adult cadavers and the morphology of the acromioclavicular joint was observed. The relevant bone structure and ligament parameters were measured, and the specimens were randomly divided into group A and group B. Group A reserved the acromioclavicular ligament and coracoclavicular ligament, and group B reserved only the acromioclavicular ligament. The difference in tension between the two groups was compared. Results The distance from the midpoint of the conical ligament to the distal end of the clavicle was (42.68 ± 6.34) mm, the width of the end point was (16.97 ± 4.28) mm, and the thickness of the center point was (5.39 ± 0.34) mm; the distance from the midpoint of the trapezoidal ligament to the clavicle was (20.35 mm ± 4.18) mm, the width of the end point was (10.35± 1.31) mm, the thickness of the center point was (5.19 ± 0.342) mm; the average vertical distance from the base of the coracoid process to the surface of the clavicle was 30.75 mm, and the mean coracoclavicular gap was 12.02 mm; the length of the central axis of the conical ligament was (15.68 ± 3.30) mm and the angle was (117.25 ± 10.80) °, while the length of the central axis of the trapezoidal ligament was (9.67 ± 2.25) mm, and the angle was (75.42± 11.37) °. The distance between the start joint of the trapezoidal ligament and the trapezium was (8.96± 3.00) mm, and the distance between the end points (13.09± 3.50) mm. The average tensile force of group A was higher than that of group B [(610.04 ± 51.24) N vs (560.41 ± 44.63) N, P < 0.05]. Conclusion During distal clavicular resection, the resection of the distal clavicle shall be within 10-30 mm. The depth shall not exceed 42 mm when drilling under the coracoid process. The reconstruction of the coracoclavicular ligament during acromioclavicular joint dislocation has an anatomical and biomechanical basis.
7.Using the external auditory canal skin graft of the healthy side to treat unilateral congenital aural atresia: preliminary results of clinical research.
Chen Long LI ; You Zhou XIE ; Ya Ying ZHU ; Yao Yao FU ; Tian Yu ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(3):338-344
Objective: To investigate whether the contralateral normal external auditory canal (EAC) skin graft can maintain the ear canal health after EAC reconstruction in unilateral congenital aural atresia (CAA) cases. Methods: A Zelen design randomized controlled study was used to collect unilateral CAA patients for EAC reconstruction prospectively (clinical trial registration number: ChiCTR2000032103). The patients were randomly divided into the control group and the trial group. The trial group used the contralateral normal EAC skin graft group (transplant part of the contralateral normal EAC skin to repair the atresia side for unilateral CAA patients), the control group all used scalp blade thick skin. We observed the EAC health and hearing results of the two groups after EAC reconstruction. Results: A total of 13 cases were enrolled from July 2020 to August 2021. There were eight patients in the trial group, including six males and two females, with an average age of 22.3 years (14-36 years). There were two patients with CAA on the left and six patients on the right. The average follow-up time was 8.8 months (4-14 months). There were five patients in the control group, all cases were male with an average age of 16.2 years (12-20 years). There were four patients with CAA on the left and one patient on the right. The average follow-up time was 7.0 months (2-14 months). In the trial group, eight cases of reconstructed EAC epithelium were healthy, one patient had cicatricial stenosis of EAC opening and lateralization of the tympanic membrane. The other patient had cicatricial stenosis of reconstructed EAC, this case also had scar hyperplasia of the contralateral EAC opening but recovered after soft packing and triamcinolone acetonide injection treatment. The healthy side EAC of the rest trial group had no scarring stenosis or local bone hyperplasia during long-term follow-up. In the control group, one patient was lost to follow-up and the other four patients had dry ears of reconstructed EAC, but easily to form crusts and needed to be cleaned repeatedly, one patient had lateralization of the tympanic membrane, the EAC epithelium was not healthy for long-term follow-up. The incidence of complications related to EAC reconstruction was lower than previous studies (χ²=5.55, P=0.018), and the average postoperative Air-Bone Gap increased (18.8±10.0)dB. Conclusion: By optimizing the EAC reconstruction technology, the health of the reconstructed EAC is improved compared with the previous study. After active intervention and treatment, there should be no scarring stenosis or local bone hyperplasia on the contralateral side EAC.
Adolescent
;
Adult
;
Ear/surgery*
;
Ear Canal/surgery*
;
Female
;
Humans
;
Male
;
Retrospective Studies
;
Skin Transplantation
;
Tympanoplasty
;
Young Adult
8.Analysis on "the essence of tuina, arrival of qi ensuring curative effect".
Meng-di XIE ; Yang LEI ; Hui-Ran YANG ; Yue YOU ; Jing-Jing GAO ; Zheng WANG ; Jie LI ; Yun-Feng ZHOU
Chinese Acupuncture & Moxibustion 2022;42(7):794-798
In reference with the systematic review of the thought of deqi (arrival of qi) put forward in Huangdi Neijing (Internal Classic of Yellow Emperor) and other classic books of traditional Chinese medicine, in view of detecting qi and identifying qi before treatment, as well as the prerequisites of deqi in tuina, meaning the accurate syndrome differentiation and manipulations, the importance of deqi in treatment with tuina is expounded. In association with clinical experience, the specific manifestations of deqi in patients during tuina are summarized, e.g. soreness, distention, pain, numbness, warm feeling and slight sweating, local changes in intestinal sound and skin color, as well as mind regulation. It is anticipated that deqi of tuina may be drawn the attention in clinical practice, and the relevant study be expanded.
Books
;
Emotions
;
Humans
;
Medicine, Chinese Traditional
;
Pain
;
Qi
9.Study on chemical constituents from fruiting bodies of Ganoderma calidophilum.
Ting-Ting ZHANG ; Jiao-Cen GUO ; Qing-Yun MA ; Fan-Dong KONG ; Li-Man ZHOU ; Qing-Yi XIE ; Hao-Fu DAI ; Zhi-Fang YU ; You-Xing ZHAO
China Journal of Chinese Materia Medica 2021;46(7):1783-1789
Chemical constituents were isolated and purified from fruiting bodies of Ganoderma calidophilum by various column chromatographic techniques, and their chemical structures were identified through combined analysis of physicochemical properties and spectral data. As a result, 11 compounds were isolated and identified as(24E)-lanosta-8,24-dien-3,11-dione-26-al(1), ganoderone A(2), 3-oxo-15α-acetoxy-lanosta-7,9(11), 24-trien-26-oleic acid(3),(23E)-27-nor-lanosta-8,23-diene-3,7,25-trione(4), ganodecanone B(5), ganoderic aldehyde A(6), 11β-hydroxy-lucidadiol(7), 3,4-dihydroxyacetophenone(8), methyl gentiate(9), ganoleucin C(10), ganotheaecolumol H(11). Among them, compound 1 is a new triterpenoid. The cytotoxic activities of all of the compounds against tumor cell lines were evaluated. The results showed that compounds 1, 3, 4 and 6 showed cytotoxic activity against BEL-7402, with IC_(50) values of 26.55, 11.35, 23.23, 18.66 μmol·L~(-1); compounds 1 and 3-6 showed cytotoxic activity against K562, with IC_(50) values of 5.79, 22.16, 12.16, 35.32, and 5.59 μmol·L~(-1), and compound 4 showed cytotoxic activity against A549, with IC_(50) value of 42.50 μmol·L~(-1).
Cell Line, Tumor
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Fruiting Bodies, Fungal
;
Ganoderma
;
Molecular Structure
;
Triterpenes/pharmacology*
10.Treatment of acetabular anterior column fracture with percutaneous screw fixation with laser-assisted axial fluoroscopy
Juzheng HU ; Zhanying SHI ; Renchong WANG ; Hao WU ; You XIE ; Chunhua MAO ; Boyu LIU ; Jingli TANG ; Chengming ZHU ; Dan ZHOU ; Bing LI
Chinese Journal of Orthopaedics 2020;40(5):302-309
Objective:To investigate the feasibility and clinical efficacy of percutaneous screw fixation for acetabular anterior column fracture with laser-assisted axial fluoroscopy.Methods:Data of 20 patients (22 sided) with acetabular anterior column fracture treated by percutaneous screw fixation with laser-assisted axial fluoroscopy from January 2017 to December 2018 were retrospectively analyzed. There were 11 males and 9 females with an average of 42.1±3.2 years (range, 24-68 years). There were 7 cases of unilateral acetabular anterior column fracture, 2 cases of bilateral acetabular anterior column fracture (4 sides), 7 cases of anterior column with ipsilateral sacral fracture, and 4 cases of anterior column with sacroiliac joint injury. There were 3 hips of Area I, 6 Area II, 13 Area III of acetabular anterior column fractures according to Nakatani partition. The time from injury to surgery was 5 days (range, 3-11 days). All patients with acetabular anterior column fractures were fixed by percutaneous screw fixation with laser-assisted axial fluoroscopy, and patients with sacral fracture or sacroiliac joint injury were fixed by percutaneous sacroiliac screws with Starr frame-assisted reduction. The time of operation, the number of intraoperative fluoroscopy and the amount of intraoperative bleeding were recorded. Matta scoring criteria were used to assess fracture reduction quality, and hip function was assessed at the last follow-up according to the modified Merle D' Aubigné and Postel scoring system.Results:The average operative time was 22±10 min (range, 20-40 min) with an average times of intraoperative fluoroscopy of 30±8 times (range, 21-45 times), and the amount of intraoperative blood loss was 20±5 ml (range, 10-40 ml). 20 patients were followed up after operation for a period of 14±3.1 months (range, 12-18 months). The quality of postoperative fracture reduction was assessed according to the Matta acetabular fracture reduction criteria: anatomical reduction in 18 hips, satisfactory reduction in 2 hips, unsatisfactory reduction in 2 hips, with an excellent and good rate of 91% (20/22). The fracture healing time was 13±2.2 weeks (range, 11-16 weeks). At the lastest follow-up, hip function was assessed according to the modified Merle D' Aubigné and Postel scoring system: excellent 18, good 3, fair 1, and the satisfactory rate was 95%(21/22). No major neurological, vascular injury, wound infection and ectopic ossification were found during follow-up.Conclusion:Using laser-assisted axial fluoroscopy percutaneous screw to treat acetabular anterior column fracture, the operation is simple. And there is low risk to damage important blood vessels and nerves. This method can shorten the operation time of acetabular anterior column fracture, reduce the amount of blood loss during the operation, and the outcome is satisfactory.

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