Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

The chemical constituents were systematically separated from the roots of Berberis polyantha by various chromatographic methods, including silica gel column chromatography, HP20 column chromatography, polyamide column chromatography, reversed-phase C_(18) column chromatography, and preparative high-performance liquid chromatography. The structures of the compounds were identified by physicochemical properties and spectroscopic techniques(1D NMR, 2D NMR, UV, MS, and CD). Four phenylpropanoids were isolated from the methanol extract of the roots of B. polyantha, and they were identified as(2R)-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone-O-β-D-glucopyranoside(1), methyl 4-hydroxy-3,5-dimethoxybenzoate(2),(+)-syringaresinol(3), and syringaresinol-4-O-β-D-glucopyranoside(4). Compound 1 was a new compound, and other compounds were isolated from this plant for the first time. The anti-inflammatory activity of these compounds was evaluated based on the release of nitric oxide(NO) in the culture of lipopolysaccharide(LPS)-induced RAW264.7 macrophages. At a concentration of 10 μmol·L~(-1), all the four compounds inhibited the LPS-induced release of NO in RAW264.7 cells, demonstrating potential anti-inflammatory properties.
Plant Roots/chemistry* ; Animals ; Mice ; Berberis/chemistry* ; RAW 264.7 Cells ; Macrophages/immunology* ; Drugs, Chinese Herbal/isolation & purification* ; Nitric Oxide/metabolism* ; Molecular Structure ; Anti-Inflammatory Agents/isolation & purification*

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

Objective:To compare the efficacy of O-arm navigation-assisted and freehand posterior pedicle screw fixation in the treatment of ankylosing spondylitis (AS) associated with lower cervical fractures in adults.Methods:A retrospective cohort study was conducted to analyze the clinical data of 29 adult AS patients with lower cervical fractures admitted to Zhengzhou Orthopedic Hospital from June 2020 to September 2024, including 22 males and 7 females, aged 35-72 years [(52.3±11.1)years]. Injured segments involved C 5~6 in 6 patients, C 6 in 8, C 6~7 in 8, and C 7 in 7. Accirdubg to AO classification, the fracture was classified as type B3 in 22 patients and type C in 7. All the patients underwent posterior cervical pedicle screw fixation, among whom 15 patients were treated with navigation-assisted screw placement (navigation screw placement group, with 120 screws) and 14 with freehand technique (freehand screw placement group, with 112 screws). The two groups were compared in terms of the operative time, intraoperative blood loss, and postoperative drainage volume. The accuracy of screw placement, penetration rate of the pedicle cortex and fracture healing time were evaluated in the two groups. The Cobb angle before surgery and at 3 days after surgery and American Spinal Injury Association (ASIA) classification before surgery and at the last follow-up were recorded in the two groups. The visual analogue scale (VAS) score, Japanese Orthopedic Association (JOA) score and neck dysfunction index (NDI) before surgery, at 1 week, 3 months after surgery and at the last follow-up were compared between the two groups. The incidence of postoperative complications was also investigated. Results:All the patients were followed up for 6-57 months [(29.3±14.1)months]. The operation duration, intraoperative blood loss, and postoperative drainage volume were (166.0±31.4)minutes, (256.7±70.7)ml, and (91.0±14.4)ml in the navigation screw placement group, which were shorter or less than (219.29±31.7)minutes, (342.1±60.7)ml, and (123.2±20.7)ml in the freehand screw placement group ( P<0.01). The accuracy of screw placement in the navigation screw placement group was 98.3% (118/120) and penetration rate of the pedicle cortex was 5.8% (7/120), which were both superior to 89.3% (100/112) and 18.8% (21/112) in the freehand screw placement group ( P<0.01). No significant difference was detected in fracture healing time between the two groups ( P>0.05). There were no significant differences in the Cobb angle before surgery and at 3 days after surgery, as well as in the ASIA grading before surgery and at the last follow-up between the two groups ( P>0.05). The Cobb angle and ASIA classification at 3 days after surgery and at the last follow-up were improved compared with those before surgery ( P<0.05). Before surgery, at 1 week, 3 months after surgery, and at the last follow-up, the VAS scores were 8.0(7.0, 9.0)points, 4.0(3.0, 4.0)points, 2.0(1.0, 2.0)points, and 0.0(0.0, 1.0)points in the navigation screw placement group, which were not significantly different from 8.0(7.0, 9.0)points, 4.0(3.0, 4.0)points, 1.5(1.0, 2.0)points, and 0.0(0.0, 1.0)points in the freehand screw placement group ( P>0.05); the JOA scores were (7.5±1.4)points, (10.9±1.2)points, (13.2±1.5)points, and (15.7±1.0)points in the navigation screw placement group, which were not significantly different from (7.2±2.8)points, (10.1±3.2)points, (11.8±3.8)points, and (14.1±4.3)points in the freehand screw placement group ( P>0.05); the NDI were 48.0(44.0, 56.0)%, 30.0(30.0, 34.0)%, 18.0(16.0, 22.0)%, 8.0(6.0, 10.0)% in the navigation screw placement group, which were not significantly different from 44.0(40.0, 52.0)%, 30.0(29.0, 34.5)%, 17.0(16.0, 20.0)%, and 8.0(5.5, 10.0)% in the freehand screw placement group ( P>0.05). The VAS scores, JOA scores, and NDI were improved at 1 week, 3 months after surgery, and at the last follow-up in both groups, compared to those before surgery and they were gradually improved with the passage of the follow-up time ( P<0.05). One patient in the navigation screw placement group developed postoperative pulmonary infection and recovered after symptomatic treatment, with an incidence of complications 7% (1/15). Two patients in the freehand screw placement group had postoperative cerebrospinal fluid leakage and dural rupture and recovered after symptomatic treatment, with an incidence of complications 14% (2/14). Throughout the follow-up period, no complications such as screw breakage, implant loosening, or loss of correction were found in either group. Conclusion:Compared with freehand screw placement, the O-arm navigation system-assisted posterior pedicle screw fixation in the treatment of AS associated with lower cervical fractures in adults demonstrates significant advantages, including reduced operative time, decreased intraoperative blood loss and postoperative drainage volume, improved accuracy of screw placement, and reduced penetration rate of the pedicle cortex.

Objective:To compare the efficacy of whole-process visualization system-assisted pedicle screw internal fixation and free-hand pedicle screw internal fixation in the treatment of thoracolumbar burst fracture (TLBF) without neurologic symptoms.Methods:A retrospective cohort study was conducted to analyze the clinical data of 64 patients with TLBF without neurologic symptoms admitted to Zhengzhou Orthopedic Hospital from December 2020 to October 2022, including 41 males and 23 females, aged 23-52 years [(42.1±6.6)years]. The injured vertebrae involved T 11 in 26 patients, T 12 in 17, L 1 in 12, and L 2 in 9. The Wiltse approach was used in all the patients, 31 of whom were treated with pedicle screw internal fixation assisted by the whole-process visualization system (visualization system-assisted screw placement group) and 33 of whom were treated with free-hand pedicle screw internal fixation (free-hand screw placement group). The two groups were compared in terms of operation time, single screw placement time, intraoperative blood loss, intraoperative total radiation dose and total length of hospital stay. The accuracy of pedicle screw placement and penetration rate of the pedicle cortex were evaluated in the two groups. The Cobb angle and lumbar visual analogue scale (VAS) before surgery, at 1 week, 3 months after surgery and at the last follow-up were compared between the two groups. The incidence of postoperative complications was also investigated. Results:All the patients were followed up for 10-33 months [(17.5±4.8)months]. The operation time was (106.9±11.8)minutes in the visualization system-assisted screw placement group, shorter than (121.3±11.4)minutes in the free-hand screw placement group ( P<0.01). The single screw placement time was (9.1±1.0)minutes in the visualization system-assisted screw placement group, shorter than (11.7±1.5)minutes in the free-hand screw placement group ( P<0.01). The total radiation dose was (10.4±2.4)mGy in the visualization system-assisted screw placement group, lower than (51.8±7.2)mGy in the screw placement group ( P<0.01). There was no significant difference in intraoperative blood loss or total length of hospital stay between the two groups ( P>0.05). The accuracy of pedicle screw placement was 96.6% (197/204) in the visualization system-assisted screw placement group, significantly higher than 89.3% (191/214) in the free-hand screw placement group ( P<0.01). Both groups showed significant improvements in Cobb angle and VAS scores at 1 week, 3 months after surgery, and at the last follow-up ( P<0.05). There were no significant differences in Cobb angle or VAS scores between the two groups at each time point ( P>0.05). In the visualization system-assisted screw placement group, one patient had incision infection at 4 days after operation, which was cured with antibiotics. One patient in the free-hand screw placement group developed the symptoms of nerve root irritation at 2 days after surgery, which disappeared at 7 days after revision. There was no significant difference in the incidence of complications between the two groups ( P>0.05). During the follow-up, no patients had broken screws, loosening of internal fixation, or loss of correction in either group. Conclusions:Compared with free-hand pedicle screw internal fixation, the whole-process visualization system-assisted pedicle screw internal fixation in the treatment of TLBF without neurologic symptoms can shorten the time of operation and screw placement, reduce the radiation dose, and improve the accuracy of pedicle screw placement, suggesting that it is a safer and more effective auxiliary method for pedicle screw placement.

Spine fracture and dislocation are common traumatic spinal conditions that often require surgical intervention due to compromised spinal stability. Surgical approaches include anterior, posterior, and combined anterior-posterior spinal procedures. According to the specific surgical requirements, patients may be placed in the prone position or repositioned between prone and supine positions during surgery. Intraoperative repositioning has become an essential step in patient positioning. However, during repositioning, patients with spinal fracture and dislocation are at increased risk for complications such as hemodynamic instability, nerve injury, and pressure injuries to the skin and soft tissue. Notably, due to the instability of the spinal cord, even minor manipulations can further exacerbate the damage, potentially leading to severe outcomes like paraplegia. Although the current clinical guidelines provide instructive recommendations for standard position, there remains no specific protocols for intraoperative repositioning in patients with spine fracture and dislocation. With a concern for the lack of clinical studies on positioning techniques, risk prevention, and operational norms for special patients, no applicable guidelines or standards are available. A consensus was required to provide clinical reference, meet the requirements of surgical treatment, and minimize the safety risks of patients caused by improper placement of positions. Professional Committee of Operating Room Nursing of Shaanxi Nursing Association organized experts in nursing management and operating room nursing from major hospitals across China to formulate Expert consensus on intraoperative repositioning for patients with spinal fracture and dislocation ( version 2025). The consensus provides 11 recommendations covering pre-repositioning preparation, intraoperative maneuvers, and post-repositioning observation, aiming to provide references for clinical standardization of the intraoperative repositioning process and protection of patients′ safety.

Objective:To compare the efficacy of three-dimensional visualization-assisted intermuscular versus conventional intermuscular cervical expansive open-door laminoplasty (CEOL) in the treatment of cervical spinal cord injury without fracture or dislocation (CSCIWFD).Methods:A retrospective cohort study was conducted to analyze the clinical data of 58 patients with CSCIWFD admitted to Zhengzhou Orthopedic Hospital from October 2021 to January 2024, including 39 males and 19 females, aged 36-77 years [(52.9±8.9)years]. Among them, 26 patients were treated with three-dimensional visualization-assisted intermuscular CEOL (three-dimensional visualization group), while 32 patients were treated with conventional intermuscular CEOL (conventional intermuscular group). All patients were treated with CEOL in 4 segments (C 3-C 6), comprising 104 surgical segments in the three-dimensional visualization group and 128 in the conventional intermuscular group. The following outcomes were compared between the two groups, including the operative duration, intraoperative blood loss, accuracy rate of open-door and hinge placement at the surgical levels, visual analogue scale (VAS) of the neck and shoulder and Japanese Orthopedic Association (JOA) score preoperatively, at 1 week, 1 month, 3 months, 6 months postoperatively and at the last follow-up, American Spinal Injury Association (ASIA) impairment scale preoperatively and at the last follow-up, and postoperative complication rate. Results:All patients were followed up for 12-24 months [(17.5±3.1)months]. The operative duration and intraoperative blood loss were (117.0±12.3)minutes and (151.3±30.9)ml in the three-dimensional visualization group, which were shorter or less than (131.9±15.0)minutes and (184.7±42.9)ml in the conventional intermuscular group ( P<0.01). The accuracy rate of open-door and hinge placement at the surgical levels was 94.2% (98/104) in the three-dimensional visualization group, significantly higher than 83.6% (107/128) in the conventional intermuscular group ( P<0.05). The VAS scores of the neck and shoulder preoperatively, at 1 week, 1 month, 3 months, 6 months postoperatively, and at the last follow-up were 6.0(5.0, 7.0)points, 3.5(3.0, 4.0)points, 3.0(2.0, 3.0)points, 2.0(1.0, 3.0)points, 2.0(1.0, 2.3)points, and 2.0(1.0, 2.0)points in the three-dimensional visualization group, which were not statistically different from 5.0(4.3, 6.8)points, 4.0(3.0, 4.0)points, 3.0(2.0, 3.0)points, 2.0(1.3, 2.0)points, 2.0(1.0, 2.8)points, and 2.0(1.0, 2.0)points in the conventional intermuscular group ( P>0.05). The VAS scores of the neck and shoulder in the two groups were significantly decreased at 1 week, 1 month and 3 months postoperatively from the preoperative values ( P<0.05), while they were stabilized at 6 months postoperatively and at the last follow-up compared with those at 3 months postoperatively, with no significant difference among them ( P>0.05). The JOA scores were (8.1±2.8)points, (10.0±2.6)points, (10.5±2.6)points, (11.6±2.3)points, (12.7±2.3)points, and (13.7±2.4)points in the three-dimensional visualization group, which were not statistically different from (8.8±2.2)points, (10.3±2.1)points, (10.8±2.0)points, (12.0±2.0)points, (12.9±2.0)points, and (13.8±2.1)points in the conventional intermuscular group ( P>0.05). The JOA scores of the two groups showed continuous improvement at 1 week, 1 month, 3 months, 6 months postoperatively and at the last follow-up in comparison with the preoperatively values ( P<0.05). Although no significant difference was observed between the two groups in ASIA grade preoperatively and at the last follow-up ( P>0.05), the ASIA grade at the last follow-up was significantly improved compared with that before surgery in both groups ( P<0.05). The postoperative complication rate was 12% (3/26) in the three-dimensional visualization group, significantly lower than 38% (12/32) in the conventional intermuscular group ( P<0.05). Conclusion:Compared with the conventional intermuscular approach, the three-dimensional visualization-assisted intermuscular CEOL offers advantages in reducing surgical trauma, improving surgical precision, and lowering the postoperative complication rate in the treatment of CSCIWFD.

Objective:To explore the value of 18F-FDG PET brain imaging in the auxiliary diagnosis of autoimmune encephalitis (AE) before treatment, and to analyze the regional and course-related characteristics of brain metabolic changes. Methods:The 18F-FDG PET brain imaging data of 49 AE patients (26 males, 23 females, age 48.0(29.0, 61.0) years) who did not receive first-line immunotherapy were retrospectively analyzed. Patients were collected from Renji Hospital, Shanghai Jiao Tong University School of Medicine, between July 2015 and December 2023. Forty-nine age- and gender-matched healthy subjects who underwent routine physical examination at the same time period were selected as the healthy controls (HC). The statistical parametric mapping (SPM) 8 two-sample t test ( P<0.001, k=50) was used to compare the imaging results of AE patients with those of HC. The screening results were adjusted by the cluster-level family-wise error rate (FWER) for P<0.05. Metabolic abnormalities associated with AE were identified, and differences in metabolic patterns at different stages of the disease course (short: ≤1 month; medium: >1 month and ≤3 month; long: >3 month) were compared by subgroup analysis. Mann-Whitney U test was used to analyze the data. Results:In the included AE patients, regions with elevated metabolism were mainly located in the limbic lobe, insula, putamen, and amygdala ( t values: 3.18-5.07, Z values: 3.17-4.76), while local metabolic reduction was observed in the frontal, parietal, and occipital lobes ( t values: 3.18-5.43, Z values: 3.23-5.06), with most of these regions passing FWER correction. In patients with anti- N-methyl- D-aspartate receptor (NMDAR) encephalitis, local metabolism increased in the right superior temporal gyrus ( t values: 3.55-4.79, Z values: 3.67-3.86) and decreased in the left middle temporal gyrus and inferior frontal gyrus ( t values: 3.55-5.43, Z values: 3.45-4.21), but the results did not pass the FWER correction. Subgroup analysis showed that in patients with short disease course ( n=17), regions with locally elevated metabolism included the brainstem, limbic lobe, and cerebellum ( t values: 3.37-5.27, Z values: 3.52-4.44), while regions with reduced metabolism were mainly located in the frontal, parietal, and occipital lobes ( t values: 3.37-6.77, Z values: 3.34-5.30), and these abnormal results all passed FWER correction. In patients with medium ( n=7) to long ( n=25) disease course, the regions with metabolic abnormalities were significantly reduced and did not pass FWER correction. Conclusions:18F-FDG PET can accurately identify brain metabolic abnormalities in AE patients, demonstrating significant regional and course-related characteristics. Metabolic abnormalities are more pronounced in patients with short disease course, while they are relatively less obvious in patients with medium to long disease course.

Objective:To investigate the relationship between collateral circulation and viable myocardium (VM) in patients with coronary chronic total occlusion (CTO).Methods:A total of 88 patients (76 males, 12 females, age (61.0±9.8) years) with coronary CTO were retrospectively analyzed. All patients underwent both 99Tc m-methoxyisobutylisonitrile (MIBI) SPECT myocardial perfusion imaging and 18F-FDG PET myocardial metabolism imaging for evaluation of VM at the First People′s Hospital of Changzhou between September 2012 and June 2023, and they were scheduled to receive coronary revascularization. The perfusion/metabolism mismatch myocardium was regarded as VM. The VM index within the CTO region was calculated, reflected the quantities of VM: VM index=(summed rest score within the CTO region-summed 18F-FDG uptake score within the CTO region)/reduced perfusion myocardial segments×4×100%. Rentrop grading of collateral circulation was performed based on coronary angiography. The differences of VM index within the CTO region between poor-developed (PD, Rentrop grade 0-1) and well-developed (WD, Rentrop grade 2-3) collateral circulation, and among different Rentrop grades were analyzed by the independent-sample t test or Kruskal-Wallis rank sum test. The linear regression analysis was used to evaluate the relationship between Rentrop grading and VM index within the CTO region. The ROC curve was constructed to analyze the predictive value of Rentrop grading for VM within the CTO region. Results:The VM index within the CTO region was significantly higher in WD patients ( n=54) compared to those in PD patients ( n=34): (45.8±16.3)% vs (21.3±16.7)% ( t=-6.79, P<0.001). Moreover, the VM index within the CTO region increased with increased Rentrop grade, and there was a significant difference among 4 groups ( H=30.22, P<0.001). Multiple linear regression analysis showed that only the Rentrop grading was an independent influencing factor for the VM index within the CTO region ( β=9.29, 95% CI: 5.91-12.67, P<0.001). ROC curve showed that the sensitivity and specificity of Rentrop grading score≥2 for predicting the presence or absence of VM within the CTO region were 65.8%(52/79) and 7/9, with the AUC of 0.724(95% CI: 0.619-0.814). Conclusions:In CTO patients who are scheduled for revascularization and evaluation of VM, as the Rentrop grading increases, the VM index within the CTO region also increases. The presence of VM within the CTO region can be predicted with Rentrop grading score ≥2.