1.Effect and mechanism of circZNF652/miR-140-3p/HMGB1 pathway on cell proliferation and migration in prostate cancer
Hua JIANG ; He ZHANG ; Songsong JIANG
Chongqing Medicine 2025;54(2):303-312,318
Objective To investigate the expression of circZNF652 in prostatic cancer(PCa)tissues and PCa cell lines,and its impact and possible mechanism on proliferation and migration.Methods Differential ex-pression of circRNAs in plasma tissues from PCa patients and benign prostatic hyperplasia(BPH)patients were detected by using gene chip technology.The expression levels of circZNF652 in PCa cell lines(22RV1,LNCaP,DU145,PC3)and normal prostate epithelial cells were measured by quantitative real-time PCR(qPCR).The correlation between different circZNF652 expression levels with the overall survival(OS)time and clinicopathological features of PCa patient were analyzed.The effects of interfering circZNF652 on PCa cell proliferation,colony formation,invasion and migration ability were assessed through CCK-8 assay,colony formation assay,Transwell assay,wound healing assay and EdU cell proliferation assay.The online database TargetScan was used to predict the binding sites between circZNF652 and miR-140-3p,and the dual-luciferase reporter gene assay and RNA pull-down assay confirmed the interaction of circZNF652 and miR-140-3p.This interaction was further validated in PC3 and DU145 cells by interfering circZNF652 as well as in blood sam-ples from PCa and BPH patients.StarBase predicted the binding sites between miR-140-3p and HMGB1,and the HMGB1 expression levels were tested after miR-140-3p overexpression in PC3 and DU145 cells.Western blot was used to detect the HMGB1 expression levels in PCa and BPH patient,PCa cell lines(DU145,PC3)as well as RWPE-1 cells.The dual-luciferase reporter gene assay,RNA immunoprecipitation(RIP)and RNA pull-down were conducted to verified the interaction between miR-140-3p and HMGB1,and the HMGB1expression in PC3 and DU145 cells were detected after transfecting different constructs(si-NC,si-cir-cZNF652#1,si-circZNF652#1+inhibitor NC,si-circZNF652#1+miR-140-3p inhibitor)to further confir-ming that miR-140-3p directly regulates HMGB1.Results The gene chip and qRT-PCR results showed that the expression level of circZNF652 in the plasma of PCa patients was higher than that in BPH patients,com-pared with low-grade PCa and BPH patients,the plasma cirZNF652 expression level in the patients with high grade PCa was upregulated(P<0.001,P<0.01).Compared with normal prostate epithelial cells,the expres-sion levels of circZNF652 in 22RV1,LNCaP,DU145 and PC3 cell lines were upregulated(P<0.05,P<0.05,P<0.001,P<0.01);The OS time in the PCa patients with circZNF652 high expression was shorter than that in PCa patients with circZNF652 low expression.Clinicopathological features analysis revealed that the circ-ZNF652 expression was correlated with the PCa stage(T stage and N stage)and Gleason score.Knocking down circZNF652 expression inhibited the proliferation,invasion and migration in DU145 and PC3 cells.The online database TargetScan predicted that there was the complementary sequences between circZNF652 and miR-140-3p,and dual-luciferase reporter gene and RNA pull-down assays confirmed thatcircZNF652 acted as a sponge for miR-140-3p in PCa cells,interfering circZNF652 experiment in PC3 and DU145 cells also proved this point.The expression of miR-140-3p in the plasma of PCa patients was significantly decreased compared with BPH patients(P<0.05),and the expression level of miR-140-3p was negatively correlate with the ex-pression level of circZNF652(r=-0.888,P<0.001).The database StarBase analysis results showed that there are complementary sequences of HMGB1 mRNA 3'-UTR and miR-140-3p,and the downstream target gene of miR-140-3p was HMGB1.The dual-luciferase reporter gene,RIP and pull-down assays confirmed the direct binding between miR-140-3p and HMGB1.The Western blot experiment found overexpression of miR-140-3p resulted in the HMGB1 expression decrease in DU145 and PC3 cells.Silencing circZNF652 expression resulted in the decrease of HMGB1 protein levels in the DU145 and PC3 cells,while inhibiting miR-140-3p could reverse this decrease.Conclusion Overexpression of circZNF652 could promote the progression and me-tastasis of PCa by regulating the miR-140-3p/HMGB1 axis.
2.Sterol regulatory element binding protein 1 promotes proliferation and invision of prostate cancer cells
Hua JIANG ; He ZHANG ; Songsong JIANG
Basic & Clinical Medicine 2025;45(2):189-196
Objective Exploring the expression of sterol regulatory element-binding protein 1(SREBP1)and acetyl-CoA carboxylase alpha(ACCα)in prostate cancer tissues and their impact on proliferation,migration and in-vasion of prostate cancer cells DU145.Methods The expression levels of SREBP1 and ACCα in prostate cancer tis?sue were analyzed using online databases including TIMER,UALCAN,GEPIA,and THE HUMAN PROTEIN ATLAS.Paraffin?embedded specimens from 58 cases of prostate cancer(PCa)and 58 cases of benign prostatic hyper?plasia(BPH)diagnosed at the Fifth Affiliated Hospital of Zunyi Medical University from January 2016 to December 2018 were collected.Immunohistochemistry was used to detect the expression of SREBP1 and ACCα in these tissues.Following the knockdown of SREBP1 in DU145 cells using shRNA,the changes in ACCα expression were assessed by qRT?PCR.The CCK?8 assay was employed to evaluate cell proliferation,while flow cytometry was used to analyze cell cycle distribution.Western blot was performed to measure the expression of SREBP1 and ACCα.Transwell assays were conducted to assess changes in migration and invasion capabilities after silencing SREBP1 in DU145 cells.Scratch assays were used to examine the impact of SREBP1 knockdown on the healing capability of DU145 cells.EdU assays were performed to detect changes in cell proliferation following the silencing of SREBP1.Oil Red O staining was utilized to observe changes in lipid content within prostate cancer cells after interference with SREBP1 expression.Results Analysis using online databases such as TIMER and UALCAN revealed that ACCα expression was signifi?cantly elevated in prostate cancer tissues compared to normal prostate tissues(P<0.001).After silencing SREBP1,the proliferation,migration and invasion capability of DU145 cells were significantly reduced compared to the control group(P<0.01),and flow cytometric analysis showed G1 phase arrested in DU145 cells(P<0.01).Knockdown of SREBP1 resulted in a significant decrease in lipid content in prostate cancer cells(P<0.01).Conclusions SREBP1 may promote the proliferation,migration and invasion of prostate cancer cells through regulating the expres?sion of ACCα.
3.Experts consensus on standard items of the cohort construction and quality control of temporomandibular joint diseases (2024)
Min HU ; Chi YANG ; Huawei LIU ; Haixia LU ; Chen YAO ; Qiufei XIE ; Yongjin CHEN ; Kaiyuan FU ; Bing FANG ; Songsong ZHU ; Qing ZHOU ; Zhiye CHEN ; Yaomin ZHU ; Qingbin ZHANG ; Ying YAN ; Xing LONG ; Zhiyong LI ; Yehua GAN ; Shibin YU ; Yuxing BAI ; Yi ZHANG ; Yanyi WANG ; Jie LEI ; Yong CHENG ; Changkui LIU ; Ye CAO ; Dongmei HE ; Ning WEN ; Shanyong ZHANG ; Minjie CHEN ; Guoliang JIAO ; Xinhua LIU ; Hua JIANG ; Yang HE ; Pei SHEN ; Haitao HUANG ; Yongfeng LI ; Jisi ZHENG ; Jing GUO ; Lisheng ZHAO ; Laiqing XU
Chinese Journal of Stomatology 2024;59(10):977-987
Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.
4.Retrospective analysis of 62 cases who received clinical application of artificial temporomandibular joint
Peng WANG ; Yao LIU ; Nan JIANG ; Ruiye BI ; Pinyin CAO ; Wenli ZHAO ; Mingjun REN ; Songsong ZHU
Chinese Journal of Stomatology 2024;59(10):1014-1018
Objective:To conduct a retrospective study on the treatment outcomes of patients who underwent artificial temporomandibular joint (TMJ) replacement surgery and to evaluate the effectiveness of artificial TMJ treatment.Methods:This study selected 62 patients who received standard Biomet artificial TMJ treatment at Department of Orthognathic and TMJ Surgery, West China Hospital of Stomatology, Sichuan University from May 2010 to September 2023 as the study subjects. Among them, there were 15 male patients and 47 female patients. The average age was 33.5 years old(ranging from 18 to 67 years). This study statistically analyzed postoperative indicators, including maximum mouth opening, forward jaw movement, lateral movement, postoperative pain scores, and patient satisfaction.Results:This study included a total of 62 patients with 99 TMJ joints. No infections occurred postoperatively. The average follow-up period was 33.7 months (ranging from 7 to 170 months). At 6 months postoperatively, the mean mouth opening was (36.1±6.2) mm, lateral movement was (2.1±0.9) mm, and forward jaw movement was (1.0±0.9) mm. The pain visual analog scale score at 6 months postoperatively was (2.8±0.6), and patient satisfaction with the surgery was (8.8±1.1). Spiral CT scans conducted after surgery showed no joint dislocation or migration, and the artificial joint remained stable during the follow-up period.Conclusions:Artificial TMJ replacement is a valuable method for effectively restoring TMJ structure and essential functions related to mouth opening and chewing. It is worthy of promotion as a reconstructive approach for the temporomandibular joint
5.Evaluation of two surgical methods for maxillary hypoplasia in patients with cleft lip and palate
Songsong GUO ; Zhenxing ZHANG ; Ping ZHANG ; Chenghui JIANG ; Jie CHENG ; Hongbing JIANG ; Sheng LI
STOMATOLOGY 2024;44(12):892-898
Objective To evaluate the clinical efficacy of anterior maxillary segmental distraction osteogenesis and Le Fort Ⅰ Osteot-omy on secondary maxillary hypoplasia in patients with cleft lip and palate.Methods Twenty-five patients with maxillary hypoplasia secondary to cleft lip and palate underwent surgery in maxillofacial surgical department of the Affiliated Stomatological Hospital of Nan-jing Medical University were selected.Ten cases were treated using tooth-borne distractors for anterior maxillary distraction osteogene-sis,and fifteen cases underwent Le Fort Ⅰ Osteotomy.Cone-beam CT,positive and lateral features,speech recording and nasopharyn-geal fibroscope were taken one week before operation and one month after surgery.The clinical outcomes of the two methods and their influence on patients'speech function were measured and compared using t-test.Results Both techniques effectively advanced the maxilla and significantly improved the patients'profiles.Anterior maxillary segmental distraction osteogenesis,which was economical and minimally invasive,greater advanced the maxilla.However,the postoperative profile was better in patients undergoing Le Fort Ⅰ osteotomy,with statistically significant differences(P<0.05).Neither technique significantly affected speech,but Le Fort Ⅰ osteotomy had a greater impact on palatopharyngeal anatomy,indicated by increased soft palate length(2.01±1.71)mm,reduced thickness(0.98±0.50)mm,and increased pharyngeal depth(3.06±1.35)mm,with statistically significant differences(P<0.05).Conclusion Anterior maxillary segmental distraction osteogenesis and orthognathic surgery are both effective methods for the treatment of secondary maxillary hypoplasia in cleft lip and palate patients.Anterior maxillary distraction osteogenesis has advantages over Le Fort I osteotomy in terms of economic benefits and its impact on the soft palate structure.
6.Orthognathic surgery and temporomandibular joint
Chinese Journal of Stomatology 2023;58(10):991-995
Orthognathic surgery changes the jaw position and occlusion, and also affects the original structure and function of the temporomandibular joint (TMJ). With the widespread development of orthognathic surgery, the impact of orthognathic surgery on the structure and function of the TMJ is increasingly valued, and the importance of the TMJ in orthognathic surgery is gradually recognized. Proper understanding the relationship between orthognathic surgery and TMJ not only helps to elucidate how the orthognathic surgery affects the condyle and causes temporomandibular disorders (TMD), but also has significant clinical significance in preventing and treating TMD in patients underwent orthognathic surgery.
7.Classification for treatment of adult temporomandibular joint ankylosis and its secondary malformations
WANG Ruiyu ; JIANG Nan ; CAO Pinyin ; LIU Yao ; ZHU Songsong ; BI Ruiye
Journal of Prevention and Treatment for Stomatological Diseases 2022;30(10):712-717
Objective:
To generate a new classification for adult temporomandibular joint ankylosis (TMJA), which could effectively guide treatment strategies for adult TMJA patients with various clinical features.
Methods:
We developed a new "CD" classification system according to the preservation of the condyle (C) and the severity of dentofacial bone deformity (D). From January 2016 to April 2020, 56 TMJ patients (with 73 ankylosed joints) in our department were classified into 4 subgroups by ‘CD’ classification: condylar head preservation but no dentofacial deformities (C+D-), no condylar head preservation and no dentofacial deformities (C-D-), condylar head preservation and dentofacial deformities (C+ D+), and no condylar head preservation but dentofacial deformities (C-D+). Different strategies were used according to the clinical features of each subgroup. The clinical outcomes of these patients were analyzed. Different treatment strategies of temporomandibular joint reconstruction were adopted for different subclasses of patients and were followed. "C +": lateral arthroplasty (LAP) was used to remove the rigidity and preserve the medial residual condyle. "C-": if the ankylosing bone ball is small and the loss of ascending branch height is not obvious, arthroplasty should be performed to relieve ankylosis; however, if the ankylosing bone ball is large and the ascending branch height decreases significantly, joint reconstruction should be carried out after the ankylosis is relieved. "D +": surgical treatment of secondary dental and maxillofacial malformations at the same time or over stages. "D-": orthodontic treatment after operation to improve occlusal relationship and symptomatic treatment of oral diseases.
Results:
After treatment, all 73 ankylosed joints were completely released, and the average maximal interincisal opening increased from (3.6±3.2 )to (32.8 ± 5.4) mm (P<0.001), with no recurrence of ankylosis found during the 12-48 month follow-up period.
Conclusion
The generation and elaboration of a ‘CD’ classification system is intended to help as a TMJA reconstruction guide for adult TMJA treatment and be widely used in more hospitals.
8.A novel biomimetic micro/nano hierarchical interface of titanium enhances adhesion, proliferation and osteogenic differentiation of bone marrow mesenchymal cells
WANG Min ; JIANG Nan ; ZHU Songsong
Journal of Prevention and Treatment for Stomatological Diseases 2021;29(4):226-233
Objective :
To design a novel biomimetic micro/nano hierarchical interface on endosseous titanium implants and investigate its effect on the biological activity of bone marrow mesenchymal cells.
Methods:
Electrochemical anodization and spark plasma sintering were used to modify smooth titanium (untreated Ti group) with a microporous trabecular bone-like architecture (micro-Ti group) and TiO2 nanotube architecture (nano-TiO2 group). Additionally, electrochemical anodization was employed to prepare TiO2 nanotubes on microporous trabecular bone-like architectures, which formed a novel biomimetic hierarchical interface (micro/nano-TiO2 group). Four groups of titanium samples were characterized by field emission scanning electron microscopy (SEM), atomic force microscopy (AFM) and contact angle (CA). Bone marrow mesenchymal cells (BMMCs) were seeded on four groups of titanium samples. Scanning electron microscopy (SEM) was employed to observe cell morphology. Cell proliferation was determined by MTT assay. The expression of focal adhesion proteins (F-actin; vinculin; osteocalcin, OCN; osteopontin, OPN) were observed under a confocal laser scanning microscope (CLSM). The mRNA expression levels of osteogenic factors (runt-related transcription factor 2, RUNX2; osteocalcin, OCN; osteopontin, OPN; collagen I, COL I) were assessed by qRT-PCR.
Results:
The micro/nano- TiO2 group featured a hydrophilic surface (CA=9° ± 2.1°). The results of the MTT assay indicated that the relative cell proliferation rates for the nano- TiO2 and micro/nano-TiO2 samples were significantly increased compared with those for the untreated-Ti and micro-Ti samples (P<0.001) after 5-9 days. The ALP results indicated that the micro/nano-TiO2 sample gained the highest value at 14 days. After 72 h of incubation, the expression of osteocalcin (OCN) and osteopontin (OPN) on micro/nano-TiO2 was the strongest. After 24 h incubation, the expression of F-actin on micro/nano-TiO2 was the strongest. In comparison with untreated-Ti and micro-Ti samples,the mRNA expression levels of all the osteogenic factors (runt-related transcription factor 2, RUNX2; osteocalcin, OCN; osteopontin, OPN; Collagen I, COL I) were markedly increased on the nano-TiO2 and micro/nano-TiO2 samples, the mRNA expression levels of collagen I (COL I) were significantly different between the nano-TiO2 and micro/nano-TiO2 samples versus the untreated-Ti and micro-Ti samples (P<0.001).
Conclusion
The novel biomimetic micro/nano hierarchical interface has a positive effect on cell attachment, viability and osteogenic differentiation of bone marrow mesenchymal cells.
9.Comprehensive measurement and quantification of bio-mechanical properties of the temporomandibular joint disc
Nan JIANG ; Yutao YANG ; Ruiye BI ; Pinyin CAO ; Yi HOU ; Songsong ZHU
Chinese Journal of Stomatology 2021;56(8):764-768
Objective:To comprehensively investigate the biomechanical properties of the temporomandibular joint (TMJ) disc and to perfect the mechanical testing system of the TMJ disc by conducting tests of compression, tension, cyclic compression, cyclic tension, creep and friction.Methods:Fifteen fresh goat heads (weighing 2.5-3.0 kg) were purchased from the market. They were all ordinary goats (9-12 months old, body weighing 18-21 kg) regardless of gender. Bilateral articular discs (a total of 30) were dissected within 30 minutes after execution. According to the national standard for mechanical testing of viscoelastic materials, fresh TMJ disc specimens of goat were prepared and tests were carried out in physiological conditions. The universal mechanical testing machine was utilized to test biomechanical properties of TMJ discs.Results:The compressive modulus of TMJ discs was (8.41±2.12) MPa and the tensile modulus was (9.54± 3.26) MPa. The mechanical characteristics would be irreversibly altered once the load exceeded the physiological range. In addition, it underwent apparent creep relaxation under continuous strain (0.5 MPa or 3.0 MPa) and the surface friction coefficient of the TMJ discs (0.015+0.011) was much lower than that of general viscoelastic materials.Conclusions:The TMJ disc was a bio-viscoelastic structure with excellent tensile and compressive properties and its surface was extremely smooth in wet conditions.
10.Clinical application of pelvic floor reconstruction in extralevator abdominoperineal excision for low rectal cancer.
Wei GE ; Gang CHEN ; Songsong JIANG ; Hao WANG
Chinese Journal of Gastrointestinal Surgery 2018;21(1):79-82
OBJECTIVETo investigate the safety and clinical significance of pelvic floor reconstruction in extralevator abdominoperineal excision(ELAPE) for advanced low rectal cancer.
METHODSThe clinical efficacy was retrospectively analyzed in 30 patients with low rectal cancer who underwent ELAPE from January 2013 to December 2016 in Nanjing Drum Tower Hospital, the affiliated Hospital of Nanjing University Medical School. There were 21 male patients and 9 female, with an average age of 61.7 years old. We used 13*15 cm Biodesign biologic meshes(Cook, China) for the reconstruction and the procedure involved soaking in saline solution for 5 minutes and fixation of the mesh to the cut edges of the levators by non-absorbable 2-0 sutures. A perineal drain was used and was removed when drainage was minimal. Potassium permanganate was used for hip bath after removing the stitches. The surgical procedure, postoperative complications, prognosis and follow-up of all these patients were documented.
RESULTSThe operations of all patients were completed successfully. ELAPE could remove more para cancer tissues in the distant rectum. There was no rectum perforation, and the circumferential resection margins of all specimens were proved to be negative. During the follow-up of 21 months, only 2 patients suffered incision infection and healed uneventfully after strengthening the dressing. No one developed perineal breakdown, bulge or intestinal obstruction, as well as local recurrence and pelvic floor hernia. There was also no complication related to mesh. The average hospitalization time was 10 days (9-15 days).
CONCLUSIONSThe ELAPE could render a low occurrence of intraoperative perforations and circumferential resection margins. Reconstruction of pelvic floor with biologic meshmight lower the complication incidences associated with the perineal region.


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