1.Indications for prenatal diagnosis using copy number variation-sequencing and detection of abnormalities: a retrospective analysis of 17 994 cases
Panlai SHI ; Yaqin HOU ; Conghui WANG ; Yanjie XIA ; Duo CHEN ; Yongchao LIU ; Junke XIA ; Li WANG ; Yin FENG ; Xiangdong KONG
Chinese Journal of Perinatal Medicine 2025;28(2):105-112
Objective:To investigate the indications for prenatal diagnosis using copy number variation-sequencing (CNV-seq) and the abnormalities detected by the method.Methods:This retrospective analysis involved 17 994 singleton pregnant women who underwent prenatal CNV-seq at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2022. These cases were divided into five groups based on the following indications for CNV-seq: abnormal fetal ultrasound findings, high-risk results indicated by non-invasive prenatal testing (NIPT) or Down's syndrome serological screening (Down's screening), adverse pregnancy history, and advanced maternal age. The proportions of cases with the indications for prenatal CNV-seq, the detection rates of abnormalities (numerical abnormalities of chromosomes, pathogenic/likely pathogenic CNV in structural abnormalities) in the five groups, and the distribution of these abnormalities were analyzed. Statistical analysis was performed using Chi-square test. Results:Among the 17 994 pregnant women, the women with abnormal fetal ultrasound findings, high-risk NIPT results, high-risk Down's screening results, adverse pregnancy history, and advanced maternal age accounted for 32.65% (5 875/17 994), 11.90% (2 142/17 994), 31.62% (5 690/17 994), 11.70% (2 105/17 994), and 12.13% (2 182/17 994), respectively. The detection rates of abnormalities in the five groups were 10.60% (623/5 875), 34.64% (742/2 142), 4.69% (267/5 690), 2.99% (63/2 105), and 3.67% (80/2 182), respectively. The overall detection rate of abnormalities was 9.86% (1 775/17 994). The cases with numerical abnormalities of chromosomes accounted for 68.79% (1 221/1 775), trisomy 21 was predominant (49.30%, 602/1 221). Chromosomal structural abnormalities were detected in 31.21% (554/1 775) of the cases with abnormalities, with 57.76% (320/554) harboring pathogenic CNVs and 42.24% (234/554) harboring likely pathogenic CNVs. The detection rate of chromosomal numerical abnormalities was higher than that of structural abnormalities in the abnormal fetal ultrasound group, NIPT high-risk group, and advanced maternal age group [6.81% (400/5 875) vs. 3.80% (223/5 875), χ2=53.10; 27.96% (599/2 142) vs. 6.68% (143/2 142), χ2=338.40; 2.43% (53/2 182) vs. 1.24% (27/2 182), χ2=8.61; all P<0.01]. A total of 416 microdeletions and 255 microduplications were detected in the 554 cases. The top three regions with the highest frequencies in microdeletions were Xp22.31 (12.74%, 53/416), 22q11.21 (7.93%, 33/416), and 17q12 (5.77%, 24/416); in microduplications, they were 22q11.21 (14.90%, 38/255), 17q12 (3.53%, 9/255), and 7q11.23 (3.53%, 9/255). Conclusions:Abnormal fetal ultrasound findings accounted for the highest proportion of prenatal diagnostic indications. The overall detection rate of abnormalities by CNV-seq is relatively high, especially in those with high-risk NIPT results as an indication for prenatal diagnosis. Among the chromosomal structural abnormalities detected in this study, the frequencies of Xp22.31 microdeletion and 22q11.21 microduplication are higher.
2.Future-Oriented High-Quality Development of Integrated Traditional Chinese and Western Medicine
Xiangmei CHEN ; Conghui WANG ; Qinxiu ZHANG ; Yan LI ; Yong TANG
Journal of Sichuan University (Medical Sciences) 2025;56(1):1-4
Integrated traditional Chinese and Western medicine refers to a clinical diagnosis and treatment model that incorporates the essence of traditional Chinese medicine in regard to treatment decisions based on syndrome differentiation and a reliance on modern western medicine as the basis of diagnosis and treatment.It is an organic combination of the two,with each bringing its own strengths into full play and complementing those of the other.After the founding of the People's Republic of China,the development of the integrated traditional Chinese and Western medicine has been increasingly valued.The unique advantages of the integration of traditional Chinese and Western medicine have been continuously utilized,resulting in fruitful achievements in theoretical inheritance and innovation and in the prevention and treatment of major diseases.Furthermore,integrated traditional Chinese and Western medicine has become a unique system of medicine of China.This article provides a review and elaboration of the entire process of the integration of traditional Chinese and Western medicine,including its significance,status of development,and achievements.It is suggested that,in the field of integrated traditional Chinese and Western medicine,efforts should made to delve into classic theories,identify the right entry points for integration,reconstruct and improve the theoretical system of integrated traditional Chinese and Western medicine by combining the cutting-edge achievements of modern medicine,and focus on enhancing clinical efficacy.
3.Erratum: Author correction to "The novel ER stress inducer Sec C triggers apoptosis by sulfating ER cysteine residues and degrading YAP via ER stress in pancreatic cancer cells" Acta Pharm Sin B 12 (2022) 210-227.
Junxia WANG ; Minghua CHEN ; Mengyan WANG ; Wenxia ZHAO ; Conghui ZHANG ; Xiujun LIU ; Meilian CAI ; Yuhan QIU ; Tianshu ZHANG ; Huimin ZHOU ; Wuli ZHAO ; Shuyi SI ; Rongguang SHAO
Acta Pharmaceutica Sinica B 2025;15(2):1208-1209
[This corrects the article DOI: 10.1016/j.apsb.2021.07.004.].
4.Mechanism of Postprandial Drowsiness and the Clinical Application under the Model of Wei Qi Circulating from Yin to Yang
Pengfei KANG ; Boju SUN ; Conghui WANG ; Xiangmei CHEN
Journal of Traditional Chinese Medicine 2025;66(8):769-774
Postprandial drowsiness is a clinical condition characterized by pronounced drowsiness after meals, while The Inner Canon of Yellow Emperor (《黄帝内经》) associates this condition with wei qi (defensive qi). By analyzing the original texts of The Inner Canon of Yellow Emperor and perspectives from many medical professionals, it is found that the transition between wakefulness and sleep depends on the mutual induction of yin and yang, and that the two pathways of wei qi circulation intersect at the spleen and stomach. Based on this, the core pathogenesis of postprandial drowsiness is proposed to be either upper jiao obstruction or spleen-stomach dysfunction, leading to the stagnation of wei qi internally, then the mutual induction of yin and yang causes inward invasion of wei qi in the body, resulting in drowsiness; at this stage, the stagnated and inward invasive wei qi converges at the spleen and stomach, then merges into the circulation of zang-fu organs, rerouting through the Foot Taiyang Meridian, forming a circulation pattern described as "circulating from yin to yang". Treatment should focus on the root and branch simultaneously, with the primary goal of regulating the circulation of wei qi; facilitating its transition from yin to yang to restore the sleep-wake cycle. By proposing the model of wei qi circulating from yin to yang, this study offers novel insights on the understanding of postprandial drowsiness.
5.Indications for prenatal diagnosis using copy number variation-sequencing and detection of abnormalities: a retrospective analysis of 17 994 cases
Panlai SHI ; Yaqin HOU ; Conghui WANG ; Yanjie XIA ; Duo CHEN ; Yongchao LIU ; Junke XIA ; Li WANG ; Yin FENG ; Xiangdong KONG
Chinese Journal of Perinatal Medicine 2025;28(2):105-112
Objective:To investigate the indications for prenatal diagnosis using copy number variation-sequencing (CNV-seq) and the abnormalities detected by the method.Methods:This retrospective analysis involved 17 994 singleton pregnant women who underwent prenatal CNV-seq at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2022. These cases were divided into five groups based on the following indications for CNV-seq: abnormal fetal ultrasound findings, high-risk results indicated by non-invasive prenatal testing (NIPT) or Down's syndrome serological screening (Down's screening), adverse pregnancy history, and advanced maternal age. The proportions of cases with the indications for prenatal CNV-seq, the detection rates of abnormalities (numerical abnormalities of chromosomes, pathogenic/likely pathogenic CNV in structural abnormalities) in the five groups, and the distribution of these abnormalities were analyzed. Statistical analysis was performed using Chi-square test. Results:Among the 17 994 pregnant women, the women with abnormal fetal ultrasound findings, high-risk NIPT results, high-risk Down's screening results, adverse pregnancy history, and advanced maternal age accounted for 32.65% (5 875/17 994), 11.90% (2 142/17 994), 31.62% (5 690/17 994), 11.70% (2 105/17 994), and 12.13% (2 182/17 994), respectively. The detection rates of abnormalities in the five groups were 10.60% (623/5 875), 34.64% (742/2 142), 4.69% (267/5 690), 2.99% (63/2 105), and 3.67% (80/2 182), respectively. The overall detection rate of abnormalities was 9.86% (1 775/17 994). The cases with numerical abnormalities of chromosomes accounted for 68.79% (1 221/1 775), trisomy 21 was predominant (49.30%, 602/1 221). Chromosomal structural abnormalities were detected in 31.21% (554/1 775) of the cases with abnormalities, with 57.76% (320/554) harboring pathogenic CNVs and 42.24% (234/554) harboring likely pathogenic CNVs. The detection rate of chromosomal numerical abnormalities was higher than that of structural abnormalities in the abnormal fetal ultrasound group, NIPT high-risk group, and advanced maternal age group [6.81% (400/5 875) vs. 3.80% (223/5 875), χ2=53.10; 27.96% (599/2 142) vs. 6.68% (143/2 142), χ2=338.40; 2.43% (53/2 182) vs. 1.24% (27/2 182), χ2=8.61; all P<0.01]. A total of 416 microdeletions and 255 microduplications were detected in the 554 cases. The top three regions with the highest frequencies in microdeletions were Xp22.31 (12.74%, 53/416), 22q11.21 (7.93%, 33/416), and 17q12 (5.77%, 24/416); in microduplications, they were 22q11.21 (14.90%, 38/255), 17q12 (3.53%, 9/255), and 7q11.23 (3.53%, 9/255). Conclusions:Abnormal fetal ultrasound findings accounted for the highest proportion of prenatal diagnostic indications. The overall detection rate of abnormalities by CNV-seq is relatively high, especially in those with high-risk NIPT results as an indication for prenatal diagnosis. Among the chromosomal structural abnormalities detected in this study, the frequencies of Xp22.31 microdeletion and 22q11.21 microduplication are higher.
6.Preliminary study on botulinum toxin type A bladder injection for the treatment of autonomic dysreflexia related to bladder dysfunction
Maping HUANG ; Hui CHEN ; Conghui HAN ; Tianhai HUANG ; Heyi ZHEN ; Xiaoyi YANG ; Qiuling LIU ; Mengxia GUO ; Hongge PAN ; Jing LIU ; Shuqing WU ; Keji XIE
Chinese Journal of Urology 2025;46(10):759-763
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.
7.Preliminary study on botulinum toxin type A bladder injection for the treatment of autonomic dysreflexia related to bladder dysfunction
Maping HUANG ; Hui CHEN ; Conghui HAN ; Tianhai HUANG ; Heyi ZHEN ; Xiaoyi YANG ; Qiuling LIU ; Mengxia GUO ; Hongge PAN ; Jing LIU ; Shuqing WU ; Keji XIE
Chinese Journal of Urology 2025;46(10):759-763
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.
8.Ancient Literature Analysis and Modern Clinical Application of Famous Classical Formula Huagaisan
Qiaonan CHEN ; Canyu SU ; Chen WANG ; Liping ZHANG ; Wei ZHOU ; Conghui WANG ; Xien LOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):35-45
ObjectiveTo sort out the historical evolution, prescription evolution and modern clinical application of Huagaisan. MethodHuagaisan and its synonym Huagaitang are used as keywords to search the databases of Traditional Chinese Medicine Think Tank, Chinese Medical Dictionary, Airusheng Chinese Medical Database and China National Knowledge Infrastructure(CNKI). According to the inclusion and exclusion criteria, we obtained the information of ancient books and modern clinical research literature related to Huagaisan, and systematically reviewed and analyzed the historical origin, prescription composition, preparation method, dosage, efficacy, medicinal material origin, processing method and modern clinical application of Huagaisan. ResultA total of 198 pieces of ancient book information were included, involving 93 ancient Chinese medicine books. Huagaisan was composed of fried Perillae Fructus, red Poria, fried Mori Cortex, Citri Eoxcarpium Rubrum, stir-fried Armeniacae Semen Amarum, Ephedrae Herba and fried Glycyrrhizae Radix et Rhizoma, which had the efficacy of promoting the lungs and relieving epidemiological symptoms, expelling phlegm and relieving cough, and treating cough with wind-cold bundled epidemiological symptoms and stagnation of phlegm and Qi. The preparation method was suggested as boiling powder, crushing the seven herbs into coarse particles, the dosage of each drug was fried Perillae Fructus of 1.27 g, red Poria of 1.27 g, fried Mori Cortex of 1.27 g, Citri Eoxcarpium Rubrum of 1.27 g, stir-fried Armeniacae Semen Amarum of 1.27 g, Ephedrae Herba of 1.27 g and fried Glycyrrhizae Radix et Rhizoma of 0.64 g, taking 8.26 g when decocting, adding 300 mL of water, decocting to 210 mL, removing the dregs, and taking it warmly after meals. Twenty-one clinical research papers were included to analyze the modern clinical application of Huagaisan, which was mainly used in the treatment of respiratory diseases such as pneumonia, asthma, bronchitis and so on. ConclusionThis paper has verified and summarized the key information of the famous classical formula Huagaisan, which can provide a detailed reference basis for the development and clinical application of its compound preparation.
9.Expression and clinical significance of FAT1 gene in pancreatic adenocarcinoma
Xinyuan LIU ; Ying YANG ; Chaodan YANG ; Zhengxiao MA ; Conghui WU ; Chen XU ; Rui ZHU ; Pan LIU ; Lisha YING ; Wenjuan YIN ; Dan SU
Chinese Journal of Oncology 2024;46(11):1029-1037
Objective:To analyze the expression of FAT1 gene in pancreatic adenocarcinoma and its relationship with clinicopathological features, prognosis, and immunotherapy for pancreatic adenocarcinoma.Methods:(1) Bioinformatics analysis: based on FAT1 mRNA expression and clinical data of 179 cases of pancreatic adenocarcinoma in the TCGA database, and FAT1 mRNA expression data of 328 cases of normal pancreatic tissues in the GTEx database. We analyzed the differences in FAT1 mRNA expression in pancreatic adenocarcinoma and normal pancreatic tissues and the relationship between FAT1 mRNA expression and the degree of differentiation, clinical stage, prognosis, immune cell infiltration, and immune checkpoint-associated genes in pancreatic adenocarcinoma. FAT1-related differentially expressed genes were analyzed by applying Limma 3.40.2 software package, and GO and KEGG enrichment analysis was performed on the differentially expressed genes. Immunohistochemical (IHC) of FAT1 in pancreatic adenocarcinoma and normal pancreatic tissues was analyzed by HPA database. (2) Validation of own tissue samples: tissue samples and clinical and prognostic data of 192 patients with pancreatic ductal adenocarcinoma admitted to Zhejiang Cancer Hospital from March 8, 2010 to September 30, 2020 were collected. IHC was performed on the tissue samples to verify the protein expression of FAT1 in pancreatic adenocarcinoma and its relationship with immune-related proteins, the degree of differentiation of pancreatic adenocarcinoma, clinical staging, and prognosis.Results:(1) Bioinformatics analysis: the FAT1 mRNA expression of 179 pancreatic adenocarcinoma tissues from the TCGA database was 5.55±1.04, which was higher than that of 328 normal pancreatic tissues with FAT1 mRNA from the GTEx database (2.95±0.53, P<0.001). FAT1-specific IHC images showed that FAT1 expression was generally high in pancreatic adenocarcinoma tissues, and FAT1 expression shifted from the cell membrane to the cytoplasm. The FAT1 mRNA expression in the highly differentiated group (31 cases), the moderately differentiated group (96 cases), and the lowly differentiated group (52 cases) were 4.99±1.46, 5.51±0.80, and 5.68±1.08, the expression of pancreatic adenocarcinoma tissues were all higher than that of normal pancreatic tissues (all P<0.001), and the FAT1 mRNA expression of the moderately differentiated group and the poorly differentiated group were all higher than that of the highly differentiated group (all P<0.001). The median progression-free survival time (PFS) and median overall survival time (OS) of the 90 patients in the FAT1 mRNA low-expression group were 16.5 and 24 months, respectively, which were longer than those of the 89 patients in the FAT1 mRNA high-expression group (median PFS and OS were 13 and 18 months, respectively; P-values were 0.011 and 0.005, respectively). Multifactorial Cox regression analysis showed that FAT1 mRNA expression level was an independent influencing factor for OS in pancreatic adenocarcinoma patients ( HR=1.47, 95% CI: 1.09-1.99). Correlation analysis showed that FAT1 mRNA expression in pancreatic adenocarcinoma was positively correlated with B-cell infiltration, CD8+ T-cell infiltration, neutrophil infiltration, macrophage infiltration, and myeloid dendritic cell infiltration ( ρ=0.27, P<0.001; ρ=0.28, P<0.001; ρ=0.32, P<0.001; ρ=0.21, P=0.004; ρ=0.32, P<0.001), and also positively correlated with mRNA expression of CD274, HAVCR2, and PDCD1LG2 ( r=0.327, P<0.001; r=0.231, P=0.002; r=0.258, P<0.001). GO and KEGG enrichment analyses showed that FAT1 mRNA expression levels were associated with activation of the Wnt signaling pathway ( P=0.029), the PI3K/Akt pathway ( P<0.001), and other tumor microenvironment-related pathways. (2) Validation of own tissue samples: among 192 pancreatic adenocarcinoma tissues, FAT1 was highly expressed in 58 cases (30.21%), and the proportion of FAT1-expressing positive tumor cells was positively correlated with the combined positive score of PD-L1 and the number of CD3+ T-cells infiltration ( r=0.154, P=0.032; r=0.287, P<0.001), and the protein expression of FAT1 had no correlation with the differentiation degree of pancreatic adenocarcinoma ( ρ=0.082, P=0.254). The median OS of 58 patients in the FAT1 high-expression group and 134 patients in the FAT1 low-expression group were 18.89 and 25.84 months, respectively, and the difference was not statistically significant (χ2=1.93, P=0.165). Conclusion:FAT1 gene is highly expressed in pancreatic adenocarcinoma tissues, may play an oncogenic role in pancreatic adenocarcinoma, may be an adverse influence on overall survival and progression-free survival of patients; FAT1 gene may be involved in multiple immune-related pathways and promote tumor immune escape.
10.Establishment of C57/B6-L and A549 cell lines stably expressing circLAMP3
Fuzai CHEN ; Conghui ZHAO ; Xiaoxuan ZHANG ; Chunping ZHANG ; Jiacheng HUANG ; Jilong CHEN ; Shujie MA
Chinese Journal of Veterinary Science 2024;44(9):2010-2016
This study aims to construct C57/B6-L and A549 cell lines that stably overexpress circu-lar RNA LAMP3(circLAMP3),laying the foundation for further research on the biological func-tions of circLAMP3.Total RNA was extracted and reverse transcripted into cDNA from C57/B6-L and A549 cells to amplify the full-length sequence of circLAMP3.Then,the fragments of cir-cLAMP3 were ligated into pLC5-ciR vector to obtain pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 recombinant plasmids.The lentiviruses expressing circLAMP3 were packaged on tran-sient transfected HEK293T cells.C57/B6-L and A549 cells were infected with lentiviruses to gen-erate cell lines overexpressing circLAMP3 through puromycin screening.To verify the overexpres-sion efficiency of circLAMP3 of cell lines,we performed the fluorescence microscopy,PCR amplifi-cation,quantitative PCR(qPCR),and Sanger sequencing experiments.The results indicated that the overexpression plasmids of pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 were suc-cessfully constructed.Strong green fluorescence was observed under a fluorescence microscopy.C57/B6-L and A549 cell lines showed a significant increase in the expression of circLAMP3 by PCR and qPCR methods.Sanger sequencing results showed that the junction site of circLAMP3 was correct.This study successfully constructed C57/B6-L and A549 cell lines overexpressing circLAMP3,providing biomaterials for further exploration of the biological function of circLAMP3 in influenza virus replication.

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