Background/Aims: Chronic constipation is prevalent yet under-diagnosed across Asia, compromising quality of life and burdening healthcare systems. Cultural stigma, varied diets, and limited access to standardized diagnostic tools delay timely care. Methods: The Bali Chronic Constipation Roundtable in November 2024, brought together experts from 11 Asian countries. The group reviewed epidemiological data, analyzed multinational questionnaire on clinical practice pattern, and conducted structured discussions to identify key barriers and propose region-specific recommendations. Results: Chronic constipation prevalence varies across Asia, ranging from 1.8% in India to 16.6% in Japan, with women and the elderly disproportionately affected. Under-reporting persists owing to cultural taboos and widespread self treatment with laxatives and traditional medications. Although the Rome IV criteria remains the global standard, they may not fully reflect Asian symptom profiles, and diagnosis is limited by scarce motility laboratories. First line therapies such as dietary-fiber optimization and osmotic laxatives are widely available, but newer pharmacotherapies (prucalopride, linaclotide, lubiprostone, and elobixibat) remain costly and unevenly accessible. Biofeedback for dyssynergic defecation is underutilized due to limited availability. Experts recommend expanded regional research on to refine diagnostic criteria, coupled with enhanced physician education and public awareness. They advocate accessibility to second-line and novel therapies that incorporate culturally attuned regional guidelines, and improved access to gastrointestinal motility testing. Conclusions The Bali Chronic Constipation Roundtable highlighted Asia’s need for region specific diagnostics and management. Addressing diagnostic and treatment gaps will improve outcomes, while ongoing researcher clinician policy collaboration must standardize guidelines, advance research, and ensure equitable care across Asia.

Background: To investigate directional deviations between pilot holes and final pedicle screw trajectories in freehand placement and analyze risk factors for misplacement. While pedicle screws are widely used in thoracolumbosacral surgery, directional deviations between pilot holes and final screw trajectories remain understudied as potential risk factors for misplacement. Methods: Thirty-three patients undergoing posterior fixation were prospectively enrolled. Inertial measurement units (IMUs) tracked the spatial positions of pedicle finders and screwdrivers via custom software. Surgeons (n = 6) were stratified into senior, middle, and junior groups. Analyzed variables included the use of tapping, surgeon experience, spinal deformity status, screw tip morphology, and surgical parameters. Results: Among 198 implanted screws (6.00 ± 2.88/patient), all exhibited trajectory deviations (8.12° ± 3.47°). Tapping significantly reduced deviations (7.23° ± 3.23° vs. 8.87° ± 3.31°, p < 0.01). Senior surgeons achieved smaller deviations (7.34° ± 3.33°) than the middle group (8.60° ± 3.51°, p = 0.039) and junior group (8.68° ± 3.49°, p = 0.022). Multivariate analysis confirmed tapping (p = 0.001) and senior experience (p = 0.023) as protective factors. No significant associations emerged for spinal deformity (8.43° ± 3.73° vs. 7.97° ± 3.35°, p = 0.377), screw tip type (cylindrical, 8.43° ± 3.26° vs. tapered, 8.07 ± 3.51°; p = 0.637), obesity, or surgical position parameters. Conclusions Freehand pedicle screw placement consistently produces trajectory deviations from pilot holes, and surgeon experience and tapping are modifiable protective factors. The IMU-based tracking system enables quantitative surgical motion analysis, suggesting its utility for training optimization and technique standardization.

The inference of tissue origin of body fluid stains is crucial for case investigation and court proceedings.However,traditional methods for identification of body fluid stains,such as morpho-logical,chemical,and immunoassay identifications have certain limitations,and there is an urgent need for more efficient methods for confirmatory experiments.In recent years,the rapid development of tran-scriptomics technology has provided new means for the identification of tissue origin of body fluid stains.Different types of RNA in the transcriptome have their own advantages.This paper elaborates in detail on the application of different types of RNA,such as mRNA,miRNA,circRNA,lncRNA,piRNA and microbial transcriptomics in body fluid identification,and summarizes their respective ad-vantages and limitations,in order to provide a reference for related research.

Objective:To evaluate the role of cannabinoid receptor 1 (CB1) in the spinal membrane in electroacupuncture (EA)-induced alleviation of morphine-triggered opioid-induced hyperalgesia (OIH) and the relationship with phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) in rats.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 2-3 months, weighing 240-260 g, in which intrathecal catheters were successfully implanted, were divided into 4 groups ( n=12 each) using a random number table method: normal saline group (NS group), morphine group (M group), morphine+ EA group (ME group), and morphine+ EA+ CB1 antagonist group (MEA group). The OIH model was established by intrathecal injection of morphine 15 μg (10 μl) twice a day for 7 consecutive days. The equal volume of normal saline 10 μl was given instead in NS group. EA of the " Yanglingquan" (GB34) and " Zusanli" (ST36) acupoints lasting 30 min was performed after the first administration of medication each day, with a current intensity of 2 mA and frequency of 2 Hz in ME group. In MEA group, morphine (15 μg) and CB1 antagonist AM251 30 μg (10 μl) were intrathecally injected twice a day for 7 consecutive days, with other treatments similar to those previously described in ME group. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before administration (T 0) and on days 1, 3, 5 and 7 after administration (T 1-4). Rats were deeply anesthetized and sacrificed at T 3, 4 after administration, and the L 4-6 spinal cord tissues were collected for determination of the expression of membrane CB1, ERK1/2 and p-ERK1/2 by Western blot. Results:Compared with NS group, the MWT was significantly decreased and the TWL was shortened at T 3, 4 in M, ME and MEA groups, the expression of spinal membrane CB1 and p-ERK1/2 was significantly up-regulated at T 4 after administration in M group, and the expression of spinal membrane CB1 was significantly up-regulated at T 3, 4 after administration in ME and MEA groups ( P<0.05). Compared with M group, the MWT was significantly increased and the TWL was prolonged at T 3, 4, and the expression of p-ERK1/2 was down-regulated at T 4 after administration in ME group ( P<0.05), no significant change was found in MWT or TWL at T 3, 4 in MEA group ( P>0.05), and the expression of spinal membrane CB1 was significantly up-regulated at T 3, 4 after administration in ME and MEA groups ( P<0.05). Compared with ME group, the MWT was significantly decreased and the TWL was shortened at T 3, 4, and the expression of spinal membrane CB1 and p-ERK1/2 was up-regulated at T 4 after administration in MEA group ( P<0.05). Conclusions:Up-regulation of spinal membrane CB1 expression is involved in EA-mediated alleviation of morphine-induced OIH, which is associated with the inhibition of ERK1/2 phosphorylation in rats.

ObjectiveA multiplex amplification system was constructed based on the capillary electrophoresis platform for simultaneous detection of saliva, semen, and vaginal secretions using tissue-specific RNA markers. The aim of this study is to identify the tissue origin of suspicious body fluid stains found at crime scenes and determine whether the body fluid stains at the crime scene are one or several types among saliva, semen, and vaginal secretions. MethodsThirty saliva samples, forty semen samples, and forty vaginal secretion samples (half from 2015 and half from 2024) were collected from healthy adult volunteers. Through primer designing, system formulation, and PCR condition optimization, a multiplex fluorescent amplification system was constructed. The specificity, sensitivity, and detection ability for mixed samples of this system were investigated, and it was tested using real crime scene materials. In the primer design stage, to reduce the requirements for RNA template quality, the amplification products were set within 80-300 bp. In the system formulation stage, dominant and subordinate primers were mainly considered. By reducing the concentration of dominant primers and increasing that of subordinate primers, a capillary electrophoresis spectrum with an appropriate peak height ratio was finally obtained. Additionally, gradient experiments were designed to adjust the concentrations of PCR reagents and PCR amplification conditions, and multiple versions of DNA amplification enzymes were optimized to achieve the best experimental results. ResultsThrough statistical analysis, there was no significant difference in the capillary electrophoresis of the 3 types of body fluid samples from the two years (2015 and 2024), demonstrating that the sample preservation method in this study can preserve samples for a relatively long time. The composite amplification system constructed in this study exhibited high specificity for all 3 types of body fluid, with no cross-reactions between the markers of each type of body fluid. The minimum detection thresholds for the 3 types of body fluid reached 0.002 9, 0.001 5, and 0.42 mg/L, respectively. This system also had a high degree of discrimination for mixed samples, especially for semen-saliva mixtures, where each body fluid marker could still be successfully detected when the concentration ratio of semen to saliva was 100:1. Meanwhile, in the two actual cases presented in this article, the application of this composite amplification system performed outstandingly. ConclusionThe composite amplification detection system constructed in this study can achieve the correct screening of saliva, semen, and vaginal secretions, overcoming the problems such as low specificity and sensitivity of marker tests and unbalanced RFU values of each marker in previous studies. The specificity and sensitivity meet the practical work requirements, and the operation is simple. It provides an analytical and identification method for body fluid stains in actual case and is applicable to the identification of the tissue origin of biological evidence at crime scenes involving sexual assault, indecent assault, and other criminal acts. In the future, more types of body fluid markers will be screened to expand the types of body fluids detected by the system, and body fluid-specific cSNP and cInDel genetic markers will be introduced to infer the sources (individuals and types) of mixed and complex stains more accurately.

Aim To explore the in vitro anti-human triple-negative breast cancer(TNBC)effect and mech-anism of Austocystin R.Methods MTT assay was used to evaluate the anti-tumor potential of Austocystin R for various human tumor cells and normal cells.Flow cytometry was employed to evaluate the influence on cell cycle progression.mRFP-GFP-LC3 adenovirus transfection was used to evaluate the autophagic flux process.Western blot assay was used to verify the effect of Austocystin R on the expression of related pro-teins.Results The results showed that Austocystin R significantly inhibited the proliferation of multiple tumor cells in a dose-dependent manner,especially for the MDA-MB-231 cells with an IC50 of 1.45μmol·L-1.In addition,Austocystin R increased the protein expression of PTEN,p53,p-p53,p27,p21,and down-regulated the expression of p-PI3K,p-AKT and p-mTOR.Austocystin R can significantly increase the proportion of S-phase MDA-MB-231 cells,inhibit the expression of Cyclin D1,CDK4,CDK6,Rb,Cyclin B1 and CDK1,and promote the expression of Cyclin E1 and CDK2.Austocystin R can promote the autophagic flux process of MDA-MB-231 cells,promote the expres-sion of LC3 Ⅰ/Ⅱ,p-Beclin-1,p-ULK1,HMGB-1 and Atg 14 proteins,and inhibit the expression of Beclin-1,ULK1,p62,ATG 3,ATG 4B,ATG 5,ATG 7,ATG 12,ATG 13 and ATG 16L1 proteins.Conclusion Austo-cystin R can exhibit its anti-TNBC activity by inhibi-ting the PI3K/AKT/mTOR signaling pathway,blocking the cell cycle at the S phase and inducing autophagic cell death.

This consensus was developed by the Orthodontic Society of the Chinese Stomatological Association to provide a systematic, scientific, and practical guideline for informed consent in orthodontic care. Orthodontic treatment is typically lengthy, highly individualized, and involves multiple factors such as growth and development, occlusal function, and facial esthetics. Rapid technological advances and diverse risk profiles make the traditional reliance on orthodontist experience or institutional templates insufficient to ensure patients′ full understanding and autonomous decision-making. To address this, the expert panel conducted extensive reviews of domestic and international guidelines, analyzed representative dispute cases, and performed multicenter patient-clinician surveys. Using a multi-round Delphi method, the group established a standardized informed consent framework covering the initial consultation, treatment, and retention phases. The consensus emphasizes that informed consent is not only a fundamental legal and ethical requirement but also a key step in building trust, improving patient compliance, and enhancing treatment satisfaction. Orthodontists should clearly and comprehensively explain treatment plans, potential risks, uncertainties, and associated costs, while respecting the autonomy of patients or guardians, and maintain continuous communication and dynamic evaluation throughout the treatment process. The release of this consensus provides unified and authoritative guidance for clinical orthodontics, helping to standardize informed consent, enhance its transparency, safeguard patient rights, reduce medical risks, and promote high-quality, sustainable development of orthodontic practice.

Objective:To evaluate the role of cannabinoid receptor 1 (CB1) in the spinal membrane in electroacupuncture (EA)-induced alleviation of morphine-triggered opioid-induced hyperalgesia (OIH) and the relationship with phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) in rats.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 2-3 months, weighing 240-260 g, in which intrathecal catheters were successfully implanted, were divided into 4 groups ( n=12 each) using a random number table method: normal saline group (NS group), morphine group (M group), morphine+ EA group (ME group), and morphine+ EA+ CB1 antagonist group (MEA group). The OIH model was established by intrathecal injection of morphine 15 μg (10 μl) twice a day for 7 consecutive days. The equal volume of normal saline 10 μl was given instead in NS group. EA of the " Yanglingquan" (GB34) and " Zusanli" (ST36) acupoints lasting 30 min was performed after the first administration of medication each day, with a current intensity of 2 mA and frequency of 2 Hz in ME group. In MEA group, morphine (15 μg) and CB1 antagonist AM251 30 μg (10 μl) were intrathecally injected twice a day for 7 consecutive days, with other treatments similar to those previously described in ME group. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before administration (T 0) and on days 1, 3, 5 and 7 after administration (T 1-4). Rats were deeply anesthetized and sacrificed at T 3, 4 after administration, and the L 4-6 spinal cord tissues were collected for determination of the expression of membrane CB1, ERK1/2 and p-ERK1/2 by Western blot. Results:Compared with NS group, the MWT was significantly decreased and the TWL was shortened at T 3, 4 in M, ME and MEA groups, the expression of spinal membrane CB1 and p-ERK1/2 was significantly up-regulated at T 4 after administration in M group, and the expression of spinal membrane CB1 was significantly up-regulated at T 3, 4 after administration in ME and MEA groups ( P<0.05). Compared with M group, the MWT was significantly increased and the TWL was prolonged at T 3, 4, and the expression of p-ERK1/2 was down-regulated at T 4 after administration in ME group ( P<0.05), no significant change was found in MWT or TWL at T 3, 4 in MEA group ( P>0.05), and the expression of spinal membrane CB1 was significantly up-regulated at T 3, 4 after administration in ME and MEA groups ( P<0.05). Compared with ME group, the MWT was significantly decreased and the TWL was shortened at T 3, 4, and the expression of spinal membrane CB1 and p-ERK1/2 was up-regulated at T 4 after administration in MEA group ( P<0.05). Conclusions:Up-regulation of spinal membrane CB1 expression is involved in EA-mediated alleviation of morphine-induced OIH, which is associated with the inhibition of ERK1/2 phosphorylation in rats.

Objective:To investigate the role and mechanism of leucine-rich α-2-glycoprotein 1 (LRG1) in the fibrosis of human pterygium fibroblasts (HPFs).Methods:A total of 30 nasal primary pterygium tissues from patients who underwent pterygium excision surgery and 30 nasal normal conjunctival tissues from patients who underwent strabismus correction surgery were collected from the First Affiliated Hospital of Harbin Medical University between January 2022 and March 2023, serving as the pterygium group and normal control group, respectively.LRG1 protein expression in both groups was detected by immunofluorescence staining.The mRNA and protein levels of LRG1 and transforming growth factor-β1 (TGF-β1) were evaluated by quantitative real-time PCR (qRT-PCR) and Western blot.Primary HPFs were cultured from excised pterygium tissues using tissue block adhesion method, and cell morphology was observed.Vmentin and cytokeratin were identified by immunofluorescence staining.HPFs were divided into recombinant human LRG1 (rhLRG1) group and blank control group treated with or without 10 μg/ml rhLRG1 for 24 hours, respectively, and cell migration was evaluated via scratch assay.Additionally, HPFs were divided into blank control group, LRG1 overexpression group and LRG1 knockdown group.HPFs in LRG1 overexpression group and LRG1 knockdown group were transfected with LRG1 overexpression plasmids and small interfering RNA for 24 hours, respectively.TGF-β1 mRNA level was evaluated by qRT-PCR and expression of TGF-β1, fibronectin (FN), type Ⅲ collagen (COL3), and α-smooth muscle actin (α-SMA) proteins were evaluated by Western blot.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Harbin Medical University (No.2022IIT026).Written informed consent was obtained from each subject.Results:HPFs were successfully isolated, exhibiting spindle-shaped morphology with whorled arrangement, positive identification for vimentin, and negative immunofluorescence staining for cytokeratin.The migration rate of the rhLRG1 group was (83.01±2.56)%, significantly higher than (50.32±4.97)% of the blank control group ( t=9.59, P<0.001).Immunofluorescence staining results showed that compared with normal conjunctival tissue, LRG1 protein was significantly higher expressed in pterygium tissue and was widely distributed in fibrous connective tissue and epithelial layer.Both mRNA and protein levels of LRG1 and TGF-β1 were significantly higher in the pterygium group than in the normal control group (mRNA: t=10.18, 6.15, both P<0.05.protein: t=6.83, 8.79, both P<0.05).In the LRG1 overexpression group, mRNA level of TGF-β1, and protein levels of FN, COL3 and α-SMA were significantly increased compared with the blank control and LRG1 knockdown groups (all P<0.05). Conclusions:LRG1 promotes fibrosis and enhances the migration ability in HPFs, and its mechanism may be associated with the upregulation of the TGF-β1 signaling pathway.

Objective:To explore the application value of coronary CT angiography(CCTA)combined with serum levels of Krueppel like factor 4(KLF4)and histone deacetylase 4(HDAC4)in assessment of coronary artery steno-sis severity in patients with coronary heart disease(CHD).Methods:A total of 152 CHD patients admitted to Suzhou High tech District People's Hospital between December 2021 and December 2023 were prospectively selected.According to coronary angiography(CAG)quantitative analysis method,patients with CHD were divided into mild stenosis group(n=57),moderate stenosis group(n=49)and severe stenosis group(n=46).Kappa consistency a-nalysis was used to determine the consistency between CCTA detection and CAG in determining the degree of coro-nary stenosis.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of CCTA combined with serum KLF4 and HDAC4 levels for the degree of coronary artery stenosis.Results:Compared to pa-tients in mild stenosis group,those in moderate stenosis group and severe stenosis group had significantly lower levels of KLF4 and HDAC4(P＜0.01 all),and those in severe stenosis group had significantly lower levels of KLF4 and HDAC4 compared to those in moderate stenosis group(P＜0.01 all).According to CCTA diagnosis,there were 53 cases with mild stenosis,53 cases with moderate stenosis and 46 cases with severe stenosis.The consistency rate with CAG diagnosis was 80.92％(123/152),which was relatively high(Kappa=0.713,P＜0.001).The accuracy of CCTA in diagnosing mild,moderate and severe stenosis was 88.16％(134/152),85.53％(130/152)and 88.16％(134/152)respectively.ROC curve analysis indicated the area under the curve(AUC)of CCTA combined with ser-um KLF4 and HDAC4 in the diagnosis of moderate and severe coronary artery stenosis was 0.949(95％CI 0.901～0.978),which was significantly higher than those of CCTA(AUC=0.867,95％CI 0.802～0.916),serum KLF4(AUC=0.895,95％CI 0.825～0.939)and HDAC4(AUC=0.809,95％CI 0.738～0.869)alone(Z=2.806,P＜0.001;Z=2.065,P=0.039;Z=3.552,P＜0.001,respectively).Conclusion:In patients with coronary heart dis-ease,serum levels of KLF4 and HDAC4 are closely related to the degree of coronary artery stenosis.The combina-tion of CCTA,serum KLF4 and HDAC4 levels has good performance in predicting the degree of coronary artery stenosis.