Objective To investigatethe relationship between gastroesophageal reflux disease (GERD) symptoms and clinicopathological characteristics, p53 expression, and survival of Chinese patients with esophageal adenocarcinoma. Methods A total of 3882 patients with esophageal adenocarcinoma, 970 matched patients with esophageal squamous cell carcinoma, and 970 matched patients with gastric cardia adenocarcinoma were included to compare the incidence of GERD symptoms. Patients with esophageal adenocarcinoma were divided into two groups according to the presence and absence of GERD symptoms. The differences in clinicopathological characteristics and p53 expression between the two groups were compared by chi-square test, and the differences in survival between the two groups were compared by landmark analysis. Results The incidence of GERD symptoms increased successively in esophageal squamous cell carcinoma, esophageal adenocarcinoma, and gastric cardia adenocarcinoma. In patients with esophageal adenocarcinoma, the proportions of male, less than 65 years old, lower thoracic tumors, tumors without squamous cell carcinoma, poorly differentiation, early tumors (stage 0-I), and p53-positive tumors in the group with GERD symptoms were higher than those in the group without GERD symptoms; moreover, the long-term survival (≥15 years) was better. Conclusion GERD symptom is an important clinical sign of esophageal adenocarcinoma. It is closely related to specific clinicopathological characteristics, p53 overexpression, and good long-term prognosis.

Objective To investigate the relationship between tooth loss and the occurrence of esophageal cancer in a natural village in Wenfeng District, Anyang City, Henan Province. Methods A prospective cohort study was conducted to observe the occurrence of tooth loss and esophageal cancer among the asymptomatic residents of the natural village for 16 years from January 2008 to July 2024. Data were analyzed by chi-square test, binary logistic regression, and restricted cubic spline. Results Among the total population of 711 cases, 136 cases were lost to follow-up and 575 cases were included in the final statistics, including 45 cases with esophageal cancer. Significant statistical difference was found between esophageal cancer patients with and without tooth loss (P<0.05). Logistic regression analysis showed that tooth loss was associated with the occurrence of esophageal cancer (OR=3.977, 95%CI: 1.543-10.255). After the adjustment for confounders, tooth loss remained significantly associated with the occurrence of esophageal cancer (OR=3.038, 95%CI: 1.035-8.914). A nonlinear dose-response relationship was observed between the number of teeth lost and the incidence of esophageal cancer. When the number of teeth lost was less than 12, the risk of esophageal cancer increased with the number of teeth lost. When more than 12 teeth were lost, the risk of esophageal cancer decreased as the number of teeth lost increased. Conclusion Tooth loss is a risk factor for the occurrence of esophageal cancer in this natural village. When the number of teeth lost is less than 12, the risk of esophageal cancer increases with the number of teeth lost.

Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there is still a lack of high-throughput methods for screening protein regulators of LLPS in target proteins. Here, we developed a CRISPR/Cas9-based screening method to identify protein phase separation regulators by integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly developed method, we screened the RNA-binding proteins that regulate PABPN1 phase separation and identified the tumor suppressor QKI as a promoter of PABPN1 phase separation. Furthermore, QKI exhibits decreased expression levels and diminished nuclear localization in colorectal cancer cells, resulting in reduced PABPN1 phase separation, which, in turn, promotes alternative polyadenylation (APA), cell proliferation, and migration in colorectal cancer.
Humans ; Colorectal Neoplasms/genetics* ; RNA-Binding Proteins/genetics* ; Poly(A)-Binding Protein I/genetics* ; CRISPR-Cas Systems ; Flow Cytometry ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

Objective:To investigate the expression and methylation status of the SALL4 gene in placental tissues of fetal growth restriction (FGR) and its effects on trophoblast cell proliferation, migration, and invasion. Methods:Placental tissues were collected from 20 full-term FGR patients and 20 healthy term controls who underwent regular prenatal examination and cesarean section at the Third Affiliated Hospital, Zhengzhou University between July 2023 and February 2024. SALL4 mRNA and protein expression were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Methylation specific polymerase china reaction (MSP) assessed promoter methylation levels. HTR8/SVneo cells were transfected with SALL4-targeting small interfering RNA (si-SALL4) or negative control small interfering RNA (si-NC). HTR8/SVneo cells were treated with the demethylating agent 5-aza-2′-deoxycytidine (5-Aza-dC) to inhibit gene methylation (5-Aza-dC group) or with 10% RPMI-1640 medium as a vehicle control. Transfection efficiency (for siRNA) and the efficacy of 5-Aza-dC-induced demethylation were assessed by qRT-PCR and Western blot. The functional effects of SALL4 knockdown and methylation inhibition on trophoblast cells were evaluated using proliferation assays, scratch wound healing assays, and Transwell invasion assays. Statistical analyses included independent t-tests and Chi-square test. Results:(1) Human tissues: FGR placentas showed lower SALL4 mRNA (0.802±0.194 vs. 1.015±0.186, t=3.55) and protein expression (0.445±0.114 vs. 0.701±0.113, t=3.19), alongside higher methylation rates of SALL4 [80% (16/20) vs. 15% (3/20), χ2=14.44] compared to controls (all P<0.05). (2) In vitro: si-SALL4 transfection reduced HTR8/SVneo proliferation (OD450 at 48 h: 0.653±0.021 vs. 0.827±0.040, t=6.60), migration [healing rate at 48 h: (24.317±2.637)% vs. (49.327±1.961)%, t=13.18], and invasion [counted invaded cells: (133.000±6.557) vs. (272.667±18.009) cells, t=12.62] versus si-NC (all P<0.05). Conversely, 5-Aza-dC treatment increased HTR8/SVneo proliferation (0.917±0.042 vs. 0.783±0.031, t=－4.47), migration [(71.097±3.354)% vs. (51.632±2.877)%, t=－7.63], and invasion [(384.000±12.166) vs. (202.833±7.095) cells, t=－13.69] versus vehicle control (all P<0.05). Conclusions:Hypermethylation of the SALL4 promoter in FGR placentas suppresses its expression, impairing trophoblast cell function. Demethylation restores SALL4 expression and enhances cellular proliferation, migration, and invasion, involving in the occurrence and development of FGR disease.

Objective:To investigate the clinical efficacy of botulinum toxin type A（BTX-A）bladder injection in the treatment of neurogenic detrusor overactivity（NDO）with autonomic dysreflexia（AD）.Methods:The patients with spinal cord injury at or above T6，who were treated at Guangdong Provincial Work Injury Rehabilitation Hospital from January 2018 to December 2022，were included in this study prospectively. Inclusion criteria：①chronic spinal cord injury patients over 18 years old（with no progression of neurological symptoms within 3 months）；② presence of NDO and AD；③ inadequate response or intolerance to oral antimuscarinic agent（M-receptor antagonists or β 3-receptor agonists）④ perform clean intermittent catheterization to empty the bladder. Exclusion criteria：① primary disease in the acute or progressive phase；② previous surgeries that would affect lower urinary tract function，such as transurethral sphincterotomy，bladder neck resection，prostatectomy，or bladder surgery；③ allergy to BTX-A or its adjuvants，or those with allergic predisposition ④ patients who were pregnant，breastfeeding，or planning for pregnancy in the near future；⑤ patients did not accept or were unable to perform intermittent catheterization. Before treatment，all patients were required to maintain 3-5 day urine diary，along with urodynamic studies（UDS），incontinence specific quality of life instrument（I-QOL）and AD symptom severity assessment，and blood pressure monitored. Key UDS parameters recorded included maximum bladder capacity，maximum detrusor pressure during filling phase，changes in maximum systolic blood pressure（SBP）relative to baseline（ΔSBP）during UDS examination，and the frequency of 24-hour blood pressure exceeding baseline by 20 mmHg. After general anesthesia or epidural anesthesia，BTX-A（200 U）was injected into the bladder at 30 points（including the triangle）under the cystoscope using a special injection needle，6.7 U per injection，and then the catheter was kept for 3-5 days after treatment. Three months later，relevant indicators were collected and compared with pre-treatment data. Results:A total of 43 patients were included in this study，including 34 males and 9 females. The age was（39.23±13.17）years old and the disease course was（2.69±3.27）years old. There were 33 cervical and 10 thoracic cases. The American Spinal Injury Association Injury Scale score distribution was as follows：26（60%）A，4（9%）B，9（21%）C，and 4（9%）D. The presence of AD was confirmed in all patients during urodynamic examination（UDS），that was the systolic blood pressure（SBP）suddenly increased and exceeded 20 mmHg（1 mmHg = 0.133 kPa）. Before treatment，The AD symptoms severity score（consist of headache，sweating，goose bumps，anxiety and palpitation）were（14.53±2.51），Bladder-related AD frequency was 10.67 episodes/day. Baseline SBP was（103.51±9.64）mmHg，the maximum SBP was（150.40±22.75）mmHg，and the change in SBP（ΔSBP）from maximum to baseline SBP during UDS examination was（43.83±21.01）mmHg. The UDS indicated that the maximum detrusor pressure during storage phase was（54.95±24.68）cmH 2O，and the bladder capacity was（131.70±75.29）ml. Bladder diary showed the volume of catheterization each time from was（181.16±49.86）ml，and The I-QOL score was（44.07±8.60）. Three months after treatment，the AD symptoms severity score（consist of headache，sweating，goose bumps，anxiety and palpitation）were（11.37±2.39）. The frequency of bladder-related AD episodes was（7.51±2.37）episodes/day，showing statistically significant differences compared to pre-treatment（ P<0.05）.The SBP before UDS examination was（102.12±10.28）mmHg，with no statistically significant difference from baseline（ P = 0.518）. The maximum SBP in perfusion phase and the ΔSBP were（132.84±16.30）mmHg and（28.72 ± 14.02）mmHg，respectively，both demonstrating statistically significant differences（ P < 0.05）. The UDS examination revealed that the maximum detrusor pressure during the storage phase was（29.77±13.72）cmH 2O，showed a significant decrease，and the bladder capacity was（272.63±79.75）ml，which were both statistically different before and after surgery. Bladder diary showed the volume of catheterization each time was（326.74±63.71）ml；I-QOL score was（71.86±11.45），both were significant different after treatment（ P < 0.01）. Conclusion:BTX-A intravesical injection in the treatment of NDO can also alleviate the severity and frequency of bladder related AD.

Objective:To investigate the clinical efficacy of botulinum toxin type A（BTX-A）bladder injection in the treatment of neurogenic detrusor overactivity（NDO）with autonomic dysreflexia（AD）.Methods:The patients with spinal cord injury at or above T6，who were treated at Guangdong Provincial Work Injury Rehabilitation Hospital from January 2018 to December 2022，were included in this study prospectively. Inclusion criteria：①chronic spinal cord injury patients over 18 years old（with no progression of neurological symptoms within 3 months）；② presence of NDO and AD；③ inadequate response or intolerance to oral antimuscarinic agent（M-receptor antagonists or β 3-receptor agonists）④ perform clean intermittent catheterization to empty the bladder. Exclusion criteria：① primary disease in the acute or progressive phase；② previous surgeries that would affect lower urinary tract function，such as transurethral sphincterotomy，bladder neck resection，prostatectomy，or bladder surgery；③ allergy to BTX-A or its adjuvants，or those with allergic predisposition ④ patients who were pregnant，breastfeeding，or planning for pregnancy in the near future；⑤ patients did not accept or were unable to perform intermittent catheterization. Before treatment，all patients were required to maintain 3-5 day urine diary，along with urodynamic studies（UDS），incontinence specific quality of life instrument（I-QOL）and AD symptom severity assessment，and blood pressure monitored. Key UDS parameters recorded included maximum bladder capacity，maximum detrusor pressure during filling phase，changes in maximum systolic blood pressure（SBP）relative to baseline（ΔSBP）during UDS examination，and the frequency of 24-hour blood pressure exceeding baseline by 20 mmHg. After general anesthesia or epidural anesthesia，BTX-A（200 U）was injected into the bladder at 30 points（including the triangle）under the cystoscope using a special injection needle，6.7 U per injection，and then the catheter was kept for 3-5 days after treatment. Three months later，relevant indicators were collected and compared with pre-treatment data. Results:A total of 43 patients were included in this study，including 34 males and 9 females. The age was（39.23±13.17）years old and the disease course was（2.69±3.27）years old. There were 33 cervical and 10 thoracic cases. The American Spinal Injury Association Injury Scale score distribution was as follows：26（60%）A，4（9%）B，9（21%）C，and 4（9%）D. The presence of AD was confirmed in all patients during urodynamic examination（UDS），that was the systolic blood pressure（SBP）suddenly increased and exceeded 20 mmHg（1 mmHg = 0.133 kPa）. Before treatment，The AD symptoms severity score（consist of headache，sweating，goose bumps，anxiety and palpitation）were（14.53±2.51），Bladder-related AD frequency was 10.67 episodes/day. Baseline SBP was（103.51±9.64）mmHg，the maximum SBP was（150.40±22.75）mmHg，and the change in SBP（ΔSBP）from maximum to baseline SBP during UDS examination was（43.83±21.01）mmHg. The UDS indicated that the maximum detrusor pressure during storage phase was（54.95±24.68）cmH 2O，and the bladder capacity was（131.70±75.29）ml. Bladder diary showed the volume of catheterization each time from was（181.16±49.86）ml，and The I-QOL score was（44.07±8.60）. Three months after treatment，the AD symptoms severity score（consist of headache，sweating，goose bumps，anxiety and palpitation）were（11.37±2.39）. The frequency of bladder-related AD episodes was（7.51±2.37）episodes/day，showing statistically significant differences compared to pre-treatment（ P<0.05）.The SBP before UDS examination was（102.12±10.28）mmHg，with no statistically significant difference from baseline（ P = 0.518）. The maximum SBP in perfusion phase and the ΔSBP were（132.84±16.30）mmHg and（28.72 ± 14.02）mmHg，respectively，both demonstrating statistically significant differences（ P < 0.05）. The UDS examination revealed that the maximum detrusor pressure during the storage phase was（29.77±13.72）cmH 2O，showed a significant decrease，and the bladder capacity was（272.63±79.75）ml，which were both statistically different before and after surgery. Bladder diary showed the volume of catheterization each time was（326.74±63.71）ml；I-QOL score was（71.86±11.45），both were significant different after treatment（ P < 0.01）. Conclusion:BTX-A intravesical injection in the treatment of NDO can also alleviate the severity and frequency of bladder related AD.

Objective:To investigate the expression and methylation status of the SALL4 gene in placental tissues of fetal growth restriction (FGR) and its effects on trophoblast cell proliferation, migration, and invasion. Methods:Placental tissues were collected from 20 full-term FGR patients and 20 healthy term controls who underwent regular prenatal examination and cesarean section at the Third Affiliated Hospital, Zhengzhou University between July 2023 and February 2024. SALL4 mRNA and protein expression were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Methylation specific polymerase china reaction (MSP) assessed promoter methylation levels. HTR8/SVneo cells were transfected with SALL4-targeting small interfering RNA (si-SALL4) or negative control small interfering RNA (si-NC). HTR8/SVneo cells were treated with the demethylating agent 5-aza-2′-deoxycytidine (5-Aza-dC) to inhibit gene methylation (5-Aza-dC group) or with 10% RPMI-1640 medium as a vehicle control. Transfection efficiency (for siRNA) and the efficacy of 5-Aza-dC-induced demethylation were assessed by qRT-PCR and Western blot. The functional effects of SALL4 knockdown and methylation inhibition on trophoblast cells were evaluated using proliferation assays, scratch wound healing assays, and Transwell invasion assays. Statistical analyses included independent t-tests and Chi-square test. Results:(1) Human tissues: FGR placentas showed lower SALL4 mRNA (0.802±0.194 vs. 1.015±0.186, t=3.55) and protein expression (0.445±0.114 vs. 0.701±0.113, t=3.19), alongside higher methylation rates of SALL4 [80% (16/20) vs. 15% (3/20), χ2=14.44] compared to controls (all P<0.05). (2) In vitro: si-SALL4 transfection reduced HTR8/SVneo proliferation (OD450 at 48 h: 0.653±0.021 vs. 0.827±0.040, t=6.60), migration [healing rate at 48 h: (24.317±2.637)% vs. (49.327±1.961)%, t=13.18], and invasion [counted invaded cells: (133.000±6.557) vs. (272.667±18.009) cells, t=12.62] versus si-NC (all P<0.05). Conversely, 5-Aza-dC treatment increased HTR8/SVneo proliferation (0.917±0.042 vs. 0.783±0.031, t=－4.47), migration [(71.097±3.354)% vs. (51.632±2.877)%, t=－7.63], and invasion [(384.000±12.166) vs. (202.833±7.095) cells, t=－13.69] versus vehicle control (all P<0.05). Conclusions:Hypermethylation of the SALL4 promoter in FGR placentas suppresses its expression, impairing trophoblast cell function. Demethylation restores SALL4 expression and enhances cellular proliferation, migration, and invasion, involving in the occurrence and development of FGR disease.

Objective:To assess respondents' understanding of the Physical Activity Questionnaire for Chronic Kidney Disease (CKD-PAQ) in Chinese and revise the items.Methods:The CKD-PAQ was sinicized according to the FACIT translation model to form a preliminary version. Using the purposive sampling method, 40 patients with chronic kidney disease from Shenzhen Traditional Chinese Medicine Hospital were selected as respondents for three rounds of cognitive interviews from July to August 2023. Targeted content analysis was used to understand the level of understanding of the initial version of the questionnaire. Based on the respondents' suggestions, the questionnaire's final version was obtained after discussing and negotiating the questionable items.Results:The CKD-PAQ contained four dimensions and 31 items. A total of 22 questions were identified in three rounds of interviews. The results of the first round of interviews show that 13 questions need to be revised in terms of understanding and expression, the results of the second round of interviews show that eight questions need to be revised in terms of understanding and expression, the results of the third round of interviews showed that the language understanding of the revised Chinese version was consistent with the original questionnaire.Conclusions:Cognitive interviews can improve respondents' understanding of the questionnaire and identify issues that are difficult to identify during the localization process, thus improving the accuracy of the questionnaire.

The unicellular green alga Chlamydomonas reinhardtii (hereafter Chlamydomonas) possesses both plant and animal attributes, and it is an ideal model organism for studying fundamental processes such as photosynthesis, sexual reproduction, and life cycle. N⁶-methyladenosine (m⁶A) is the most prevalent mRNA modification, and it plays important roles during sexual reproduction in animals and plants. However, the pattern and function of m⁶A modification during the sexual reproduction of Chlamydomonas remain unknown. Here, we performed transcriptome and methylated RNA immunoprecipitation sequencing (MeRIP-seq) analyses on six samples from different stages during sexual reproduction of the Chlamydomonas life cycle. The results show that m⁶A modification frequently occurs at the main motif of DRAC (D = G/A/U, R = A/G) in Chlamydomonas mRNAs. Moreover, m⁶A peaks in Chlamydomonas mRNAs are mainly enriched in the 3' untranslated regions (3'UTRs) and negatively correlated with the abundance of transcripts at each stage. In particular, there is a significant negative correlation between the expression levels and the m⁶A levels of genes involved in the microtubule-associated pathway, indicating that m⁶A modification influences the sexual reproduction and the life cycle of Chlamydomonas by regulating microtubule-based movement. In summary, our findings are the first to demonstrate the distribution and the functions of m⁶A modification in Chlamydomonas mRNAs and provide new evolutionary insights into m⁶A modification in the process of sexual reproduction in other plant organisms.
Animals ; Chlamydomonas reinhardtii/metabolism* ; Reproduction/genetics* ; Life Cycle Stages/genetics* ; Transcriptome ; Plants/genetics*

Objective : To explore the relationship between the methylation level of RABL6 in peripheral blood and early lung cancer (LC) with a case-control study in the Chinese population． Methods : The methylation levels of 7 CpG sites in RABL6 gene in peripheral blood of samples from 275 LC patients (81．5% at stage I) ，and age- and gender-matched 185 benign lung nodule cases and 267 matched healthy controls were measured by matrix-assisted laser desorption ionization time-of-flight mass spectrometry．Multinomial Logistic regression adjusted for covariates was used to analyze the association between the RABL6 methylation and LC．Mann-Whitney U test was applied for the comparisons of RABL6 methylation levels between clinical characteristics subgroups of LC. Results : Compared to the healthy controls，the methylation of RABL6 _CpG_ 17 was inversely associated with LC in females ( per - 10% methylation : OR = 2. 47，95% CI = 1. 19－5. 13，P = 0. 016) ，but positively associated with LC in males (per - 10% methylation : OR = 0. 52，95% CI = 0. 29－0. 94，P = 0. 030) ．In addition，hypermethylation of RABL6_CpG _2 and RABL6_CpG_5 was significantly associated with LC in the subjects older than 55 years (for RABL6_CpG_ 2 : per －10% methylation : OR = 0. 77，95% CI = 0. 60－0. 99，P = 0. 038 ; for RABL6_CpG_5 : OR = 0. 58，95% CI = 0. 34－0. 97，P = 0. 038) ． Conclusion The study reveals an association between peripheral blood-based RABL6 methylation levels and early LC，providing a new clue for developing peripheral blood-based DNA methylation as a potential marker for the evaluation of LC risk.