1.Assessing traditional Chinese medicine from a molecular perspective
Minhui Su ; Wenxi Wang ; Xudong Sun ; Lihong Li ; Weihong Tan
Journal of Traditional Chinese Medical Sciences 2025;2025(1):3-9
Molecular medicine, which delves into the intricacies of biomolecular structure, function, and role, is pivotal for advancing precise diagnostics and personalized treatment. Nucleic acids, a class of star functional molecules, are notable for their versatile applications in molecular diagnostics, gene therapy, and drug development. Therefore, in this study, we review the extensive use of nucleic acid aptamers in medicinal practice. Furthermore, the expanding field of molecular medicine has catalyzed advancements in traditional Chinese medicine (TCM), as evidenced by scientific endeavors to integrate modern technologies. Therefore, TCM has experienced rapid modernization by leveraging artificial intelligence, nucleic acid molecular medicine, and bioelectronic medicine.
2.Correlations of insomnia severity with cognitive memory, depression and anxiety in patients with chronic insomnia
Xingyu RAN ; Yuxi LIU ; Chen SUN ; Wenxi LUO ; Weineng CHEN ; Fengjuan SU ; Fuping XU ; Zhong PEI
Chinese Journal of Neuromedicine 2025;24(2):147-153
Objective:To explore the correlative factors for insomnia severity in chronic insomnia patients using MemTrax memory test.Methods:Two hundred and twenty-two chronic insomnia patients (insomnia≥3 days per week with a duration≥3 months) recruited from Center for Preventive Treatment of Diseases, Guangdong Provincial Hospital of Chinese Medicine or through in-hospital advertisements from April 2024 to September 2024 were chosen. Pittsburgh Sleep Quality Index (PSQI) was used to evaluate the sleep quality over the last month; according to PSQI score, these patients were divided into mild insomnia group (scores of 7-10), moderate insomnia group (scores of 11-15) and severe insomnia group (scores of 16-21). MemTrax memory test was used to record the picture recognition accuracy and picture recognition reaction time, and MemTrax comprehensive index (MTx-Cp) was calculated; Patients' Health Questionnaire (PHQ-9), and Generalized Anxiety Disorder-7 scale(GAD-7) were used to evaluate the depression and anxiety status of these patients in recent 2 weeks. The clinical data, MemTrax test results, PHQ-9 and GAD-7 scores of patients with different degrees of chronic insomnia were compared. Spearman rank correlation analysis was used to investigate the correlation between insomnia severity and clinical data such as cognitive memory function in chronic insomnia patients.Results:Among the 220 chronic insomnia patients, 54 had mild insomnia, 111 had moderate insomnia, and 55 had severe insomnia. Severe insomnia patients had significantly higher percentages of those>50 years old and those using hypnotics compared with mild insomnia patients and moderate insomnia patients ( P<0.05). Compared with the mild insomnia patients and moderate insomnia patients, the severe insomnia patients exhibited significantly lower picture recognition accuracy (90%[86%, 94%], 88%[82%, 94%], 84%[78%, 92%]), significantly lower MTx-Cp (88.55±18.67, 84.41±20.93, 76.69±17.43), and significantly higher PHQ-9 score (9[6, 11], 9[6, 15], 12[8, 16], P<0.05). Moreover, severe insomnia patients had significantly longer picture recognition reaction time and higher GAD-7 score than mild insomnia patients (1.11[1.03, 1.24] s vs. 1.04[0.90, 1.15] s; 7[5, 13] vs. 6[3, 9], P<0.05). Spearman rank correlation analysis showed that insomnia severity in chronic insomnia patients was positively correlated with age, PHQ-9 score, GAD-7 score, and picture recognition reaction time, and negatively correlated with picture recognition accuracy and MTx-Cp ( P<0.05). Conclusion:Insomnia severity in patients with chronic insomnia is correlated with age, cognitive memory function, depression and anxiety.
3.Expert consensus on pulpotomy in the management of mature permanent teeth with pulpitis.
Lu ZHANG ; Chen LIN ; Zhuo CHEN ; Lin YUE ; Qing YU ; Benxiang HOU ; Junqi LING ; Jingping LIANG ; Xi WEI ; Wenxia CHEN ; Lihong QIU ; Jiyao LI ; Yumei NIU ; Zhengmei LIN ; Lei CHENG ; Wenxi HE ; Xiaoyan WANG ; Dingming HUANG ; Zhengwei HUANG ; Weidong NIU ; Qi ZHANG ; Chen ZHANG ; Deqin YANG ; Jinhua YU ; Jin ZHAO ; Yihuai PAN ; Jingzhi MA ; Shuli DENG ; Xiaoli XIE ; Xiuping MENG ; Jian YANG ; Xuedong ZHOU ; Zhi CHEN
International Journal of Oral Science 2025;17(1):4-4
Pulpotomy, which belongs to vital pulp therapy, has become a strategy for managing pulpitis in recent decades. This minimally invasive treatment reflects the recognition of preserving healthy dental pulp and optimizing long-term patient-centered outcomes. Pulpotomy is categorized into partial pulpotomy (PP), the removal of a partial segment of the coronal pulp tissue, and full pulpotomy (FP), the removal of whole coronal pulp, which is followed by applying the biomaterials onto the remaining pulp tissue and ultimately restoring the tooth. Procedural decisions for the amount of pulp tissue removal or retention depend on the diagnostic of pulp vitality, the overall treatment plan, the patient's general health status, and pulp inflammation reassessment during operation. This statement represents the consensus of an expert committee convened by the Society of Cariology and Endodontics, Chinese Stomatological Association. It addresses the current evidence to support the application of pulpotomy as a potential alternative to root canal treatment (RCT) on mature permanent teeth with pulpitis from a biological basis, the development of capping biomaterial, and the diagnostic considerations to evidence-based medicine. This expert statement intends to provide a clinical protocol of pulpotomy, which facilitates practitioners in choosing the optimal procedure and increasing their confidence in this rapidly evolving field.
Humans
;
Calcium Compounds/therapeutic use*
;
Consensus
;
Dental Pulp
;
Dentition, Permanent
;
Oxides/therapeutic use*
;
Pulpitis/therapy*
;
Pulpotomy/standards*
4.Impact of intensive blood pressure lowering on atrial fibrillation risk in hypertensive patients: A systematic review and meta-analysis
Wenxi ZUO ; Yuhe HUANG ; Ziyi SUN ; Yuhan YANG ; Jin ZHANG ; Xiaoxiao ZHANG ; Kuiwu YAO
Science of Traditional Chinese Medicine 2025;3(2):186-193
Background: Hypertension is a major risk factor for cardiovascular diseases, including AF, which is one of the most common cardiac arrhythmias globally. AF is strongly associated with an increased risk of stroke, heart failure (HF), and cardiovascular mortality. Although intensive blood pressure lowering has been shown to reduce adverse cardiovascular events, its effect on the risk of AF remains debated. Some studies suggest a beneficial effect, whereas others are inconclusive. Therefore, a comprehensive review and meta-analysis are needed to clarify these effects. Objective: This study aims to evaluate the impact of intensive blood pressure lowering on the incidence of atrial fibrillation (AF) in hypertensive patients. Methods: We performed a systematic review and meta-analysis by searching PubMed, EMBASE, Scopus, Web of Science, and the Cochrane Library up to September 2, 2024, for randomized controlled trials comparing intensive blood pressure lowering with standard treatment in hypertensive patients. Studies were included if participants were 40 year or older with systolic blood pressure between 130 and 180 mm Hg (1 mm Hg≈0.133 kPa). Data extraction was conducted by 2 independent researchers, and statistical analysis was performed using Review Manager (RevMan) 5.4. Risk ratios (RRs) and 95% confidence intervals (CIs) were calculated. A random-effects model was applied if heterogeneity was detected (I
> 50%). Results: A total of 6 randomized controlled trials involving 34,824 participants were included in the analysis. Intensive blood pressure lowering significantly reduced the risk of new-onset AF compared with standard treatment (RR = 0.76, 95% CI = 0.62-0.93, p < 0.01, I
= 0%). Reductions were also observed in stroke (RR = 0.71, 95% CI = 0.58-0.87, p < 0.005, I
= 7%), HF (RR = 0.67, 95% CI = 0.45-0.99, p = 0.05, I
= 53%), and nonfatal coronary events (RR = 0.80, 95% CI = 0.70-0.92, p < 0.005, I
= 39%). However, intensive blood pressure lowering had no significant effect on cardiovascular mortality or all-cause mortality compared with standard treatment. Discussion: Intensive blood pressure lowering significantly reduces the risk of AF and other cardiovascular events, such as stroke, HF, and nonfatal coronary events, particularly among high-risk hypertensive patients. These findings support the potential benefits of intensive blood pressure management in reducing AF incidence and improving overall cardiovascular outcomes, but the evidence is limited.
5.Expression of SAPCD2 in esophageal squamous cell carcinoma tissues and its effects on the biological function of esophageal squamous cell carcinoma cells in vitro and the possible mechanisms
Qixiong LONG ; Jingchun LI ; Junxia FENG ; Wenxi HE ; Wei SUN
Cancer Research and Clinic 2025;37(5):334-342
Objective:To investigate the expression of SAPCD2 in esophageal squamous cell carcinoma (ESCC) tissues and its effects on the biological function of ESCC cells in vitro, as well as the possible molecular mechanisms.Methods:By using the Gene Expression Profile Interaction Analysis (GEPIA) platform, the transcriptional level expressions of SAPCD2 in 182 ESCC samples and 286 normal esophageal tissue samples from the Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx) database were analyzed. Immunohistochemical (IHC) staining was performed on clinical tissue chips of ESCC patients, and staining scores were evaluated. The expression differences of SAPCD2 protein between 61 cancer tissues and the paired adjacent tissues with the complete clinical data, as well as the distribution of patients with SAPCD2 high expression among patients stratifies by different clinicopathological features were compared. ESCC cell line KYSE-150 was transfected with plasmids carrying SAPCD2 sequence and short hairpin RNA sequence targeting SAPCD2, respectively, which was treated as the SAPCD2 overexpression group and SAPCD2 knockdown group; and the cells transfected with empty plasmids and plasmids carrying negative RNA sequence were treated as the overexpression control group and the knockdown control group. CCK-8 method (expressed with the absorbance value) and plate clone formation assay were used to detect the cell proliferation ability. Cell migration was detected by using cell scratch assay and Transwell cell migration assay were used to detect the cell migration ability. The reverse-real time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the mRNA levels of SAPCD2, matrix metalloproteinase-9 (MMP9), proliferating cell nuclear antigen (PCNA), and Vimentin in cells of all groups. Western blotting was used to detect the expression levels of SAPCD2 protein and proteins related to cell proliferation, invasion, metastasis, and AKT signaling pathway.Results:GEPIA platform analysis showed that the transcriptional expression level of SAPCD2 in ESCC tissues was higher than that in adjacent tissues, and the difference was statistically significant ( P < 0.01). IHC staining showed that the staining score of SAPCD2 protein in cancer tissues was higher than that in adjacent tissues [(8.2±2.8) points vs. (2.2±1.7) points], and the proportion of patients with positive SAPCD2 protein (staining score > 0 point) in cancer tissues was higher than that in adjacent tissues [95.1% (58/61) vs. 57.4% (35/61)], and the differences were statistically significant (all P < 0.001), while there were no statistically significant differences in the distribution of the high expression of SAPCD2 protein (staining score > 3 points) patients stratified by gender, age, tumor size, pathological grade, and T stage, N stage and M stage (all P > 0.05). CCK-8 assay showed that the absorbance value of KYSE-150 cells in the SAPCD2 overexpression group after 96 h of culture was higher than that in the overexpression control group, while the absorbance values of the SAPCD2 knockdown group after 72 h and 96 h of culture were lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). The plate clone formation assay showed that the number of colonies of KYSE-150 cells cultured for 14 d in the SAPCD2 overexpression group was more than that in the overexpression control group [(800±30) vs. (458±47)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(52±7) vs. (81±2)], and the differences were statistically significant (all P < 0.05). The cell scratch assay showed that after 24 h of culture, the scratch width of KYSE-150 cells in the SAPCD2 overexpression group was narrower than that in the overexpression control group [(51±9) μm vs. (89±7) μm], while that in the SAPCD2 knockdown group was wider than that in the knockdown control group [(120±22) μm vs. (37±10) μm], and the differences were statistically significant (all P < 0.01). Transwell cell migration assay showed that the migration number of KYSE-150 cells in the SAPCD2 overexpression group was more than that in the overexpression control group [(202±18) vs. (50±14)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(227±27) vs. (483±16)], and the differences were all statistically significant (all P < 0.01). qPCR assay showed that the mRNA relative expression levels of MMP9, PCNA and Vimentin in KYSE-150 cells in the SAPCD2 overexpression group were all higher than those in the overexpression control group, while those in the SAPCD2 knockdown group were all lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). Western blotting showed that the relative expression levels of PCNA and Vimentin proteins in KYSE-150 cells of the SAPCD2 overexpression group were higher than those of the overexpression control group, while the relative expression levels of epithelial cadherin (E-cad) and cleaved cysteine aspartate protease 3 (CASP3) proteins were lower than those of the overexpression control group; however, the expression levels in SAPCD2 knockdown group showed the opposite results, and the differences were statistically significant (all P < 0.05); the relative expression level of phosphorylated AKT (p-AKT) protein in the SAPCD2 overexpression group was higher than that in the overexpression control group, while that in the SAPCD2 knockdown group was lower than that in the knockdown control group, and the differences were statistically significant (all P < 0.01). Conclusions:SAPCD2 is highly expressed at both the transcriptional level and the protein level in ESCC tissues. SAPCD2 promotes the proliferation and migration of ESCC cells in vitro, which may be related to the AKT signaling pathway.
6.Analysis of the therapeutic effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery,and intersphincteric resection surgery in the treatment of low rectal cancer
Wenxi LI ; Xin ZHENG ; Baoxin SUN ; Haisheng ZHANG ; Zhida ZHU ; Enhong ZHAO
Journal of Clinical Surgery 2025;33(6):632-636
Objective To investigate the effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery(NOSES),and intersphincteric resection(ISR)on treatment outcomes and quality of life in patients with low rectal cancer.Methods A total of 152 patients with low rectal cancer who were admitted from January 2020 to June 2022,and they were divided into the traditional laparoscopic group(49 cases),the NOSES group(51 cases),and the ISR group(52 cases)according to the surgical method.The operation status,postoperative recovery status,pain,anal function recovery status,quality of life and complications were compared in the 3 groups.Results The operation time of the traditional laparoscopic group[(193.98±12.31)min]was lower than that of the NOSES group[(203.54±15.02)min]and the ISR group[(199.85±11.98)min](P<0.05),operation time of NOSES group and ISR group was no difference(P>0.05).The first exhaust time[(60.21±10.05)h],the first time of getting out of bed[(37.52±6.21)h],and the length of postoperative hospital stay[(12.51±1.47)d]in the traditional laparoscopic group were all higher than those in the NOSES group[(51.06±8.67)h,(30.13±4.92)h,and(11.27±)1.23)d]and ISR group[(53.19±9.24)h,(28.97±4.71)h,(11.73±1.35)d](P<0.05),and there were no statistically significant differences in the first exhaust time,the first time to get out of bed,and the length of postoperative hospital stay between the NOSES and ISR groups(P>0.05).There was no statistically significant difference in the Visual Analogue Scale(VAS)scores for pain at 4 hours,24 hours,and 48 hours after surgery among the three groups(P>0.05).The VAS scores of the three groups at 24 hours after surgery were higher than those at 4 hours and 48 hours after surgery,and the difference was statistically significant(P<0.05).The VAS scores of the three groups at 48 hours after surgery were higher than those at 4 hours after surgery,and the difference was statistically significant(P<0.05).The NOSES group's Wexner score[(4.93±0.76)points]at 3 months after surgery and Wexner score[(3.21±0.42)points]at 6 months after surgery were lower than those of the ISR group[(6.32±0.93)points,(4.48±0.54)points]and the traditional laparoscopic group[(5.93±0.81)points,(4.01±0.53)points](P<0.05),and the Wexner score of the 3 groups at 3 months after surgery was lower than that at 1 month after surgery(P<0.05).The EORTC QLQ-C30 score of the NOSES group at 3 months after surgery was(74.82±4.01)points,and that at 6 months was(85.49±4.93)points,which were higher than those of the ISR group[(67.05±5.03)points and(71.64±4.21)points]and the traditional laparoscopic group[(70.42±3.92)points,(76.28±4.48)points](P<0.05),and the EORTC QLQ-C30 scores of the traditional laparoscopic group at 3 and 6 months after surgery were higher than those of the ISR group,and the difference was statistically significant(P<0.05).The EORTC QLQ-C30 score of the 3 groups at 6 months after surgery was higher than that before surgery and 3 months after surgery(P<0.05),and the EORTC QLQ-C30 score of the 3 groups at 3 months after surgery was higher than that before surgery(P<0.05).There was no significant difference in the incidence of total complications among the three groups(P>0.05).Conclusion Compared with traditional laparoscopic surgery for low rectal cancer,the NOSES and ISR methods accelerate postoperative bowel function recovery,and the NOSES methods have advantages in anal function recovery and better and satisfactory quality of life.
7.Analysis of the therapeutic effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery,and intersphincteric resection surgery in the treatment of low rectal cancer
Wenxi LI ; Xin ZHENG ; Baoxin SUN ; Haisheng ZHANG ; Zhida ZHU ; Enhong ZHAO
Journal of Clinical Surgery 2025;33(6):632-636
Objective To investigate the effects of traditional laparoscopic surgery,natural orifice specimen extraction surgery(NOSES),and intersphincteric resection(ISR)on treatment outcomes and quality of life in patients with low rectal cancer.Methods A total of 152 patients with low rectal cancer who were admitted from January 2020 to June 2022,and they were divided into the traditional laparoscopic group(49 cases),the NOSES group(51 cases),and the ISR group(52 cases)according to the surgical method.The operation status,postoperative recovery status,pain,anal function recovery status,quality of life and complications were compared in the 3 groups.Results The operation time of the traditional laparoscopic group[(193.98±12.31)min]was lower than that of the NOSES group[(203.54±15.02)min]and the ISR group[(199.85±11.98)min](P<0.05),operation time of NOSES group and ISR group was no difference(P>0.05).The first exhaust time[(60.21±10.05)h],the first time of getting out of bed[(37.52±6.21)h],and the length of postoperative hospital stay[(12.51±1.47)d]in the traditional laparoscopic group were all higher than those in the NOSES group[(51.06±8.67)h,(30.13±4.92)h,and(11.27±)1.23)d]and ISR group[(53.19±9.24)h,(28.97±4.71)h,(11.73±1.35)d](P<0.05),and there were no statistically significant differences in the first exhaust time,the first time to get out of bed,and the length of postoperative hospital stay between the NOSES and ISR groups(P>0.05).There was no statistically significant difference in the Visual Analogue Scale(VAS)scores for pain at 4 hours,24 hours,and 48 hours after surgery among the three groups(P>0.05).The VAS scores of the three groups at 24 hours after surgery were higher than those at 4 hours and 48 hours after surgery,and the difference was statistically significant(P<0.05).The VAS scores of the three groups at 48 hours after surgery were higher than those at 4 hours after surgery,and the difference was statistically significant(P<0.05).The NOSES group's Wexner score[(4.93±0.76)points]at 3 months after surgery and Wexner score[(3.21±0.42)points]at 6 months after surgery were lower than those of the ISR group[(6.32±0.93)points,(4.48±0.54)points]and the traditional laparoscopic group[(5.93±0.81)points,(4.01±0.53)points](P<0.05),and the Wexner score of the 3 groups at 3 months after surgery was lower than that at 1 month after surgery(P<0.05).The EORTC QLQ-C30 score of the NOSES group at 3 months after surgery was(74.82±4.01)points,and that at 6 months was(85.49±4.93)points,which were higher than those of the ISR group[(67.05±5.03)points and(71.64±4.21)points]and the traditional laparoscopic group[(70.42±3.92)points,(76.28±4.48)points](P<0.05),and the EORTC QLQ-C30 scores of the traditional laparoscopic group at 3 and 6 months after surgery were higher than those of the ISR group,and the difference was statistically significant(P<0.05).The EORTC QLQ-C30 score of the 3 groups at 6 months after surgery was higher than that before surgery and 3 months after surgery(P<0.05),and the EORTC QLQ-C30 score of the 3 groups at 3 months after surgery was higher than that before surgery(P<0.05).There was no significant difference in the incidence of total complications among the three groups(P>0.05).Conclusion Compared with traditional laparoscopic surgery for low rectal cancer,the NOSES and ISR methods accelerate postoperative bowel function recovery,and the NOSES methods have advantages in anal function recovery and better and satisfactory quality of life.
8.Correlations of insomnia severity with cognitive memory, depression and anxiety in patients with chronic insomnia
Xingyu RAN ; Yuxi LIU ; Chen SUN ; Wenxi LUO ; Weineng CHEN ; Fengjuan SU ; Fuping XU ; Zhong PEI
Chinese Journal of Neuromedicine 2025;24(2):147-153
Objective:To explore the correlative factors for insomnia severity in chronic insomnia patients using MemTrax memory test.Methods:Two hundred and twenty-two chronic insomnia patients (insomnia≥3 days per week with a duration≥3 months) recruited from Center for Preventive Treatment of Diseases, Guangdong Provincial Hospital of Chinese Medicine or through in-hospital advertisements from April 2024 to September 2024 were chosen. Pittsburgh Sleep Quality Index (PSQI) was used to evaluate the sleep quality over the last month; according to PSQI score, these patients were divided into mild insomnia group (scores of 7-10), moderate insomnia group (scores of 11-15) and severe insomnia group (scores of 16-21). MemTrax memory test was used to record the picture recognition accuracy and picture recognition reaction time, and MemTrax comprehensive index (MTx-Cp) was calculated; Patients' Health Questionnaire (PHQ-9), and Generalized Anxiety Disorder-7 scale(GAD-7) were used to evaluate the depression and anxiety status of these patients in recent 2 weeks. The clinical data, MemTrax test results, PHQ-9 and GAD-7 scores of patients with different degrees of chronic insomnia were compared. Spearman rank correlation analysis was used to investigate the correlation between insomnia severity and clinical data such as cognitive memory function in chronic insomnia patients.Results:Among the 220 chronic insomnia patients, 54 had mild insomnia, 111 had moderate insomnia, and 55 had severe insomnia. Severe insomnia patients had significantly higher percentages of those>50 years old and those using hypnotics compared with mild insomnia patients and moderate insomnia patients ( P<0.05). Compared with the mild insomnia patients and moderate insomnia patients, the severe insomnia patients exhibited significantly lower picture recognition accuracy (90%[86%, 94%], 88%[82%, 94%], 84%[78%, 92%]), significantly lower MTx-Cp (88.55±18.67, 84.41±20.93, 76.69±17.43), and significantly higher PHQ-9 score (9[6, 11], 9[6, 15], 12[8, 16], P<0.05). Moreover, severe insomnia patients had significantly longer picture recognition reaction time and higher GAD-7 score than mild insomnia patients (1.11[1.03, 1.24] s vs. 1.04[0.90, 1.15] s; 7[5, 13] vs. 6[3, 9], P<0.05). Spearman rank correlation analysis showed that insomnia severity in chronic insomnia patients was positively correlated with age, PHQ-9 score, GAD-7 score, and picture recognition reaction time, and negatively correlated with picture recognition accuracy and MTx-Cp ( P<0.05). Conclusion:Insomnia severity in patients with chronic insomnia is correlated with age, cognitive memory function, depression and anxiety.
9.Expression of SAPCD2 in esophageal squamous cell carcinoma tissues and its effects on the biological function of esophageal squamous cell carcinoma cells in vitro and the possible mechanisms
Qixiong LONG ; Jingchun LI ; Junxia FENG ; Wenxi HE ; Wei SUN
Cancer Research and Clinic 2025;37(5):334-342
Objective:To investigate the expression of SAPCD2 in esophageal squamous cell carcinoma (ESCC) tissues and its effects on the biological function of ESCC cells in vitro, as well as the possible molecular mechanisms.Methods:By using the Gene Expression Profile Interaction Analysis (GEPIA) platform, the transcriptional level expressions of SAPCD2 in 182 ESCC samples and 286 normal esophageal tissue samples from the Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx) database were analyzed. Immunohistochemical (IHC) staining was performed on clinical tissue chips of ESCC patients, and staining scores were evaluated. The expression differences of SAPCD2 protein between 61 cancer tissues and the paired adjacent tissues with the complete clinical data, as well as the distribution of patients with SAPCD2 high expression among patients stratifies by different clinicopathological features were compared. ESCC cell line KYSE-150 was transfected with plasmids carrying SAPCD2 sequence and short hairpin RNA sequence targeting SAPCD2, respectively, which was treated as the SAPCD2 overexpression group and SAPCD2 knockdown group; and the cells transfected with empty plasmids and plasmids carrying negative RNA sequence were treated as the overexpression control group and the knockdown control group. CCK-8 method (expressed with the absorbance value) and plate clone formation assay were used to detect the cell proliferation ability. Cell migration was detected by using cell scratch assay and Transwell cell migration assay were used to detect the cell migration ability. The reverse-real time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the mRNA levels of SAPCD2, matrix metalloproteinase-9 (MMP9), proliferating cell nuclear antigen (PCNA), and Vimentin in cells of all groups. Western blotting was used to detect the expression levels of SAPCD2 protein and proteins related to cell proliferation, invasion, metastasis, and AKT signaling pathway.Results:GEPIA platform analysis showed that the transcriptional expression level of SAPCD2 in ESCC tissues was higher than that in adjacent tissues, and the difference was statistically significant ( P < 0.01). IHC staining showed that the staining score of SAPCD2 protein in cancer tissues was higher than that in adjacent tissues [(8.2±2.8) points vs. (2.2±1.7) points], and the proportion of patients with positive SAPCD2 protein (staining score > 0 point) in cancer tissues was higher than that in adjacent tissues [95.1% (58/61) vs. 57.4% (35/61)], and the differences were statistically significant (all P < 0.001), while there were no statistically significant differences in the distribution of the high expression of SAPCD2 protein (staining score > 3 points) patients stratified by gender, age, tumor size, pathological grade, and T stage, N stage and M stage (all P > 0.05). CCK-8 assay showed that the absorbance value of KYSE-150 cells in the SAPCD2 overexpression group after 96 h of culture was higher than that in the overexpression control group, while the absorbance values of the SAPCD2 knockdown group after 72 h and 96 h of culture were lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). The plate clone formation assay showed that the number of colonies of KYSE-150 cells cultured for 14 d in the SAPCD2 overexpression group was more than that in the overexpression control group [(800±30) vs. (458±47)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(52±7) vs. (81±2)], and the differences were statistically significant (all P < 0.05). The cell scratch assay showed that after 24 h of culture, the scratch width of KYSE-150 cells in the SAPCD2 overexpression group was narrower than that in the overexpression control group [(51±9) μm vs. (89±7) μm], while that in the SAPCD2 knockdown group was wider than that in the knockdown control group [(120±22) μm vs. (37±10) μm], and the differences were statistically significant (all P < 0.01). Transwell cell migration assay showed that the migration number of KYSE-150 cells in the SAPCD2 overexpression group was more than that in the overexpression control group [(202±18) vs. (50±14)], and that in the SAPCD2 knockdown group was less than that in the knockdown control group [(227±27) vs. (483±16)], and the differences were all statistically significant (all P < 0.01). qPCR assay showed that the mRNA relative expression levels of MMP9, PCNA and Vimentin in KYSE-150 cells in the SAPCD2 overexpression group were all higher than those in the overexpression control group, while those in the SAPCD2 knockdown group were all lower than those in the knockdown control group, and the differences were statistically significant (all P < 0.05). Western blotting showed that the relative expression levels of PCNA and Vimentin proteins in KYSE-150 cells of the SAPCD2 overexpression group were higher than those of the overexpression control group, while the relative expression levels of epithelial cadherin (E-cad) and cleaved cysteine aspartate protease 3 (CASP3) proteins were lower than those of the overexpression control group; however, the expression levels in SAPCD2 knockdown group showed the opposite results, and the differences were statistically significant (all P < 0.05); the relative expression level of phosphorylated AKT (p-AKT) protein in the SAPCD2 overexpression group was higher than that in the overexpression control group, while that in the SAPCD2 knockdown group was lower than that in the knockdown control group, and the differences were statistically significant (all P < 0.01). Conclusions:SAPCD2 is highly expressed at both the transcriptional level and the protein level in ESCC tissues. SAPCD2 promotes the proliferation and migration of ESCC cells in vitro, which may be related to the AKT signaling pathway.
10.Development of a
Yiying YANG ; Qingqing SUN ; Yang LIU ; Hanzhi YIN ; Wenping YANG ; Yang WANG ; Ying LIU ; Yuxian LI ; Shen PANG ; Wenxi LIU ; Qian ZHANG ; Fang YUAN ; Shiwen QIU ; Jiong LI ; Xuefeng WANG ; Keqiang FAN ; Weishan WANG ; Zilong LI ; Shouliang YIN
Journal of Zhejiang University. Science. B 2021;22(5):383-396


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