Objective:To investigate the effect and potential mechanism of circular RNA-centrosome and spindle pole-associated protein 1 (circ-CSPP1) on the malignant biological behavior of hepatoma cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the expressions of circ-CSPP1 and microRNA-582-5p (miR-582-5p) in hepatoma cells, and Western blotting was used to detect the expression of karyopherins α2 (KPNA2). HepG2 cells were divided into the circ-CSPP1 overexpression group, the circ-CSPP1 overexpression control group, the si-CSPP1 group, the si-NC group, the si-CSPP1+ miR-582-5p inhibition group, and the si-CSPP1+ miR-582-5p inhibition control group. circ-CSPP1 overexpression plasmid, CSPP1 interfering small RNA, CSPP1 interfering small RNA, miR-582-5p inhibition sequence and negative control were transfected respectively in these groups. Cell proliferation in each group was detected by 5-acetylene-2'-deoxyuridine (Edu), invasion ability was detected by Transwell assay, and the binding of circ-CSPP1 and KPNA2 to miR-582-5p was verified by dual-luciferase assay. In the si-CSPP1 group, HepG2 cells transfected with si-CSPP1 lentivirus were subcutaneously injected into the back of nude mice ( n=12), and in the si-NC group, HepG2 cells transfected with negative control lentivirus ( n=12) were injected. The tumor mass, volume, circ-CSPP1 and KPNA2 were detected. Results:In the circ-CSPP1 overexpression group, the relative expression of circ-CSPP1 was (1.68±0.17), the expression of KPNA2 was (1.52±0.16), and the number of invasive cells in the 100-fold field of view was (128.4±13.5), which were all higher than those in the circ-CSPP1 overexpression control group [(1.25±0.16), (1.24±0.15), (128.4±13.5)], while the expression of miR-552-5p was lower than that in the circ-CSPP1 overexpression control group [(0.96±0.11) vs (1.31±0.15)]; The relative expression of circ-CSPP1 in the si-CSPP1 group was (1.02±0.13), KPNA2 was (0.74±0.09), and the number of invasive cells was (53.5±6.7), which were lower than those in the si-NC group [(1.28±0.14), (1.22±0.13), (74.6±8.3)], while the expression of miR-582-5p was higher than that in the si-NC group [(1.71±0.18) vs (1.32±0.14)]; The expression of circ-CSPP1 and KPNA2 and the number of invasion cells in the si-CSPP1+ miR-582-5p inhibition group was higher than that in the si-CSPP1+ miR-582-5p inhibition control group, and the differences were statistically significant (all P<0.05). The results of cell proliferation were consistent with those of invasion. The dual-luciferase gene report showed that, compared with the miR-NC group, the relative luciferase activity in HepG2 cells co-transfected with circ-CSPP1-WT or KPNA2-WT wild-type reporter vectors in the miR-882-5p mimic group decreased [(0.46±0.05) vs (1.03±0.11), (0.42±0.03) vs (1.01±0.09)]. The differences were all statistically significant (both P<0.05). However, there was no statistically significant difference in the relative luciferase activity in HepG2 cells co-transfected with the circ-CSPP1-MUT or KPNA2-MUT mutant reporter vectors (both P>0.05). The tumor weight, volume and circ-CSPP1 and KPNA2 expressions in tumor tissue of nude mice in the si-CSPP1 group were all lower than those in the si-NC group, and the expression of miR-582-5p was higher than that in the si-NC group. The differences were statistically significant (all P<0.05). Conclusion:Inhibition of circ-CSPP1 suppressed the malignant biological behavior of hepatoma cells and tumor growth by upregulating miR-582-5p and downregulating KPNA2.

Objective:To investigate the effect and potential mechanism of circular RNA-centrosome and spindle pole-associated protein 1 (circ-CSPP1) on the malignant biological behavior of hepatoma cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the expressions of circ-CSPP1 and microRNA-582-5p (miR-582-5p) in hepatoma cells, and Western blotting was used to detect the expression of karyopherins α2 (KPNA2). HepG2 cells were divided into the circ-CSPP1 overexpression group, the circ-CSPP1 overexpression control group, the si-CSPP1 group, the si-NC group, the si-CSPP1+ miR-582-5p inhibition group, and the si-CSPP1+ miR-582-5p inhibition control group. circ-CSPP1 overexpression plasmid, CSPP1 interfering small RNA, CSPP1 interfering small RNA, miR-582-5p inhibition sequence and negative control were transfected respectively in these groups. Cell proliferation in each group was detected by 5-acetylene-2'-deoxyuridine (Edu), invasion ability was detected by Transwell assay, and the binding of circ-CSPP1 and KPNA2 to miR-582-5p was verified by dual-luciferase assay. In the si-CSPP1 group, HepG2 cells transfected with si-CSPP1 lentivirus were subcutaneously injected into the back of nude mice ( n=12), and in the si-NC group, HepG2 cells transfected with negative control lentivirus ( n=12) were injected. The tumor mass, volume, circ-CSPP1 and KPNA2 were detected. Results:In the circ-CSPP1 overexpression group, the relative expression of circ-CSPP1 was (1.68±0.17), the expression of KPNA2 was (1.52±0.16), and the number of invasive cells in the 100-fold field of view was (128.4±13.5), which were all higher than those in the circ-CSPP1 overexpression control group [(1.25±0.16), (1.24±0.15), (128.4±13.5)], while the expression of miR-552-5p was lower than that in the circ-CSPP1 overexpression control group [(0.96±0.11) vs (1.31±0.15)]; The relative expression of circ-CSPP1 in the si-CSPP1 group was (1.02±0.13), KPNA2 was (0.74±0.09), and the number of invasive cells was (53.5±6.7), which were lower than those in the si-NC group [(1.28±0.14), (1.22±0.13), (74.6±8.3)], while the expression of miR-582-5p was higher than that in the si-NC group [(1.71±0.18) vs (1.32±0.14)]; The expression of circ-CSPP1 and KPNA2 and the number of invasion cells in the si-CSPP1+ miR-582-5p inhibition group was higher than that in the si-CSPP1+ miR-582-5p inhibition control group, and the differences were statistically significant (all P<0.05). The results of cell proliferation were consistent with those of invasion. The dual-luciferase gene report showed that, compared with the miR-NC group, the relative luciferase activity in HepG2 cells co-transfected with circ-CSPP1-WT or KPNA2-WT wild-type reporter vectors in the miR-882-5p mimic group decreased [(0.46±0.05) vs (1.03±0.11), (0.42±0.03) vs (1.01±0.09)]. The differences were all statistically significant (both P<0.05). However, there was no statistically significant difference in the relative luciferase activity in HepG2 cells co-transfected with the circ-CSPP1-MUT or KPNA2-MUT mutant reporter vectors (both P>0.05). The tumor weight, volume and circ-CSPP1 and KPNA2 expressions in tumor tissue of nude mice in the si-CSPP1 group were all lower than those in the si-NC group, and the expression of miR-582-5p was higher than that in the si-NC group. The differences were statistically significant (all P<0.05). Conclusion:Inhibition of circ-CSPP1 suppressed the malignant biological behavior of hepatoma cells and tumor growth by upregulating miR-582-5p and downregulating KPNA2.

Objective:To analyze the influencing factors of low liver regeneration in patients with hilar cholangiocarcinoma (HCCA) after portal vein embolization (PVE).Method:Clinical data of 62 patients with HCCA undergoing PVE at Henan Provincial People's Hospital (People's Hospital of Zhengzhou University) from January 2019 to March 2024 were retrospectively analyzed, including 33 males and 29 females, aged (59.1±10.3) years. Patients were divided into two groups based on the median regeneration rate of remnant liver volume (28.6%) three weeks after PVE: low regeneration ( n=31, <28.6%) and high regeneration group ( n=31, ≥28.6%). The proportion of lymph node metastasis, history of alcohol consumption, liver fibrosis, biliary tract infection, alkaline phosphatase (ALP), and tumor necrosis factor-α (TNF-α) were compared between two groups. Multivariate logistic regression analysis was used to indentify the influencing factors of low liver regeneration in patients with HCCA after PVE surgery. Results:The proportion of lymph node metastasis, history of alcohol consumption, liver fibrosis, biliary tract infection, ALP, and level of TNF-α were higher in the low regeneration group than those in the high regeneration group (all P<0.05). Multivariate logistic regression analysis showed that patients with regional lymph node metastasis ( OR=2.561, 95% CI: 1.265-5.185), history of alcohol consumption ( OR=2.616, 95% CI: 1.321-5.181), liver fibrosis ( OR=2.351, 95% CI: 1.265-4.369), biliary tract infection ( OR=2.461, 95% CI: 1.226-4.940), elevated level of ALP ( OR=2.687, 95% CI: 1.351-5.344), and elevated level of TNF-α ( OR=2.781, 95% CI: 1.452-5.326) had an increased risk of low liver regeneration after PVE (all P<0.05). Conclusion:Regional lymph node metastasis, history of alcohol consumption, liver fibrosis, biliary tract infection, and elevated ALP and TNF-α are risk factors for low liver regeneration in patients with HCCA after PVE surgery, which should be noted in clinical practice.

Morels (Morchella spp.) are valuable medicinal and edible mushrooms. In this study, chemical profiles of aromas and lipophilic extracts of black morel (Morchella importuna) grown in China were analyzed by gas chromatography/mass spectroscopy, along with the evaluation of antioxidant and antimicrobial activities for the lipophilic extracts. Sixty-five compounds in total were identified from the aromas, and 1-octen-3-ol was the main component for aromas of fresh (34.40%) and freeze-dried (68.61%) black morels, while the most abundant compound for the aroma of the oven-dried sample was 2(5H)-furanone (13.95%). From the lipophilic extracts, 29 compounds were identified with linoleic acid as the main compound for fresh (77.37%) and freeze-dried (56.46%) black morels and steroids (92.41%) as the main constituent for an oven-dried sample. All three lipophilic extracts showed moderate antioxidant activities against 2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with the IC50 values ranging 7.56 ∼ 17.52 mg/mL and 5.75 ∼ 9.73 mg/mL, respectively, and no obvious antimicrobial activity was observed for lipophilic extracts. The drying methods affect the chemical profile of black morel, and freeze-drying was favorable for retaining nutrients and morel smell. This is the first report on the aroma and lipophilic extracts of M. importuna grown in China.

Objective:To explore the optimization strategy of the Asia-Pacific colorectal screening (APCS) scoring system in the screening of colorectal neoplasms.Methods:From February to Decomber in 2016 and March to December in 2018, at Xijing Hospital of Air Force Military Medical University and the First Affiliated Hospital of Xi′an Medical University, patients who received opportunistic screening colonoscopy were enrolled. Before colonoscopy, the APCS score (low-risk zero to one points, medium-risk two to three points and high-risk four to seven points), body mass index (BMI), fecal occult blood test (FOBT) and plasma methylated Septin9 gene ( mSEPT9) of all patients were detected and recorded. The results of colonoscopy and biopsy pathology were taken as the gold standard, the efficacies of the above methods in screening colorectal neoplasms were compared to determine and optimize the screening efficiency of APCS scoring system. Chi-square test was used for statistical analysis. Results:A total of 494 patients were screened, of whom 133 cases were diagnosed with colorectal polyps, including 86 cases of colorectal adenomatous polyps (82 cases of non-progressive adenoma, and four cases of advanced-adenoma), and 47 cases of non-adenomatous polyps. According to the APCS score, the detection rate of colorectal adenomatous polyps of the high-risk group (33.3%, 33/99) was 2.02 and 3.76 times higher than those of the medium-risk group (16.5%, 39/237) and low-risk group (8.9%, 14/158), respectively (both Bonferroni correction test, both P<0.016). The detection rate of colorectal adenomatous polyps of patients with BMI>23.9 kg/m 2 was significantly higher than that of patients with BMI≤23.9 kg/m 2 (22.2%, 59/266 vs. 11.8%, 27/228), and the difference was statistically significant ( χ2=9.126, P=0.003). There was no statistically significant difference in the detection rate of colorectal adenomatous polyps between patients with positive- mSEPT9 expression and patients with negative- mSEPT9 expression (22.4%, 15/67 vs. 17.3%, 47/271) ( χ2=0.913, P=0.378). Among 158 low and medium risk patients (APCS score≤three points) who underwent simultaneous BMI measurement, FOBT and plasma mSEPT9 test, the detection rate of colorectal adenomatous polyps in patients with BMI>23.9 kg/m 2 was higher than that in patients with BMI≤23.9 kg/m 2 (17.8%, 16/90 vs. 5.9%, 4/68), and the difference was statistically significant ( χ2=4.957, P=0.030). The redetection efficacy of colorectal adenomatous polyps in patients with BMI>23.9 kg/m 2 and FOBT-positive was higher than that in patients with BMI≤23.9 kg/m 2 and FOBT-negative (28.1%, 9/32 vs. 8.0%, 4/50) and the difference was statistically significant ( χ2=5.942, P=0.027). In addition, the redetection rate of colorectal adenomatous polyps of patients with positive expression of FOBT and plasma mSEPT9 was also higher than that of patients with negative expression (5/14 vs. 12.9%, 12/93), and the difference was statistically significant ( χ2=4.738, P=0.045). Conclusions:When the APCS scoring system is used for sequential screening of colorectal tumors, the optinal choice of BMI replacement or combined with FOBT can improve the patients′ compliance and screening efficiency, which has significant clinical significance and promotion value in the early diagnosis and treatment of colorectal neoplasms.

Objective:To investigate the relationship between the expression of DNA methyltransferase 3b (DNMT3b) and the methylation of SEPT9 gene, and their application prospects in the diagnosis and treatment of colorectal cancer.Methods:Seventy-five cases of colorectal cancer and adjacent tissues, 68 cases of colorectal high-grade internal neoplasia tissues (referred to as precancerous tissues) and high-grade internal adjacent neoplasia tissues (referred to as adjacent precancerous tissues) were collected. Pyrosequencing was used to detect the methylationlevel of SETP9. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressionof SEPT9 and immunohistochemistry(IHC) was applied to detect the protein expressions of SETP9 and DNMT3b. Liposome-mediated method was used to transfect DNMT3b siRNA and negative control siRNA into HT-29 cells. Five groups including DNMT3b siRNA 15 nmol/L group, DNMT3b siRNA 30 nmol/L group, negative control siRNA 15 nmol/L group, negative control siRNA 30 nmol/L group and blank control group were set up. Pyrosequencing was applied to determine the methylation level of SEPT9 and mRNA expression of DNMT3b in each group.Results:The methylation rates of SEPT9 gene in colorectal cancer tissues, adjacent tissues, precancerous tissues and adjacent precancerous tissues were (76.8±6.5)%, (14.4±2.6)%, (34.6±5.0)% and (7.4±1.2)%, respectively, which was highest in colorectal cancer tissue ( P<0.001). The relative expressions of SEPT9 mRNA were 0.18±0.03, 0.89±0.41, 0.69±0.41 and 1.01±0.21, respectively, which was lowest in colorectal cancer tissue ( P<0.001), while there were no statistically significant differences in adjacent tissues, precancerous tissues and adjacent precancerous tissues ( P>0.05). The positive rates of SEPT9 protein expression were 12.0% (9/75), 53.3% (40/75), 55.1% (38/69) and 62.3% (43/69), which was lowest in the colorectal cancer tissue ( P<0.001), while there were no statistically significant differences in the adjacent group, precancerous group and adjacent precancerous group ( P>0.016 7). The positive rates of DNMT3b protein expression were 56.3% (45/75), 26.7% (20/75), 46.4% (32/69) and 33.3% (23/69), respectively, which was highest in colorectal cancer tissue ( P<0.001), while without statistically significant difference from the precancerous tissue ( P>0.016 7). Experiments in vitro showed that DNMT3b mRNA expression was lowest in DNMT3b siRNA 30 nmol/L group among five groups and was statistically different from other groups (all P<0.05). Meanwhile, the methylationrate of SEPT9 gene was lowest in this group, but without statistically significant difference from the DNMT3b siRNA 15 nmol/L group ( P>0.05). Conclusions:The expression of DNMT3b is significantly correlated with the methylation level of SEPT9 gene in different stages of colorectal cancer. The high expression of DNMT3b may be an important molecular event before SEPT9 gene methylation and it may have an important potential application value in the diagnosis and treatment of early colorectal cancer.

Objective:To investigate the relationship between the expression of DNA methyltransferase 3b (DNMT3b) and the methylation of SEPT9 gene, and their application prospects in the diagnosis and treatment of colorectal cancer.Methods:Seventy-five cases of colorectal cancer and adjacent tissues, 68 cases of colorectal high-grade internal neoplasia tissues (referred to as precancerous tissues) and high-grade internal adjacent neoplasia tissues (referred to as adjacent precancerous tissues) were collected. Pyrosequencing was used to detect the methylationlevel of SETP9. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressionof SEPT9 and immunohistochemistry(IHC) was applied to detect the protein expressions of SETP9 and DNMT3b. Liposome-mediated method was used to transfect DNMT3b siRNA and negative control siRNA into HT-29 cells. Five groups including DNMT3b siRNA 15 nmol/L group, DNMT3b siRNA 30 nmol/L group, negative control siRNA 15 nmol/L group, negative control siRNA 30 nmol/L group and blank control group were set up. Pyrosequencing was applied to determine the methylation level of SEPT9 and mRNA expression of DNMT3b in each group.Results:The methylation rates of SEPT9 gene in colorectal cancer tissues, adjacent tissues, precancerous tissues and adjacent precancerous tissues were (76.8±6.5)%, (14.4±2.6)%, (34.6±5.0)% and (7.4±1.2)%, respectively, which was highest in colorectal cancer tissue ( P<0.001). The relative expressions of SEPT9 mRNA were 0.18±0.03, 0.89±0.41, 0.69±0.41 and 1.01±0.21, respectively, which was lowest in colorectal cancer tissue ( P<0.001), while there were no statistically significant differences in adjacent tissues, precancerous tissues and adjacent precancerous tissues ( P>0.05). The positive rates of SEPT9 protein expression were 12.0% (9/75), 53.3% (40/75), 55.1% (38/69) and 62.3% (43/69), which was lowest in the colorectal cancer tissue ( P<0.001), while there were no statistically significant differences in the adjacent group, precancerous group and adjacent precancerous group ( P>0.016 7). The positive rates of DNMT3b protein expression were 56.3% (45/75), 26.7% (20/75), 46.4% (32/69) and 33.3% (23/69), respectively, which was highest in colorectal cancer tissue ( P<0.001), while without statistically significant difference from the precancerous tissue ( P>0.016 7). Experiments in vitro showed that DNMT3b mRNA expression was lowest in DNMT3b siRNA 30 nmol/L group among five groups and was statistically different from other groups (all P<0.05). Meanwhile, the methylationrate of SEPT9 gene was lowest in this group, but without statistically significant difference from the DNMT3b siRNA 15 nmol/L group ( P>0.05). Conclusions:The expression of DNMT3b is significantly correlated with the methylation level of SEPT9 gene in different stages of colorectal cancer. The high expression of DNMT3b may be an important molecular event before SEPT9 gene methylation and it may have an important potential application value in the diagnosis and treatment of early colorectal cancer.

Objective: To compare the clinical effects between laparoscopic cholecystectomy and choledochotomy versus traditional open cholecystectomy plus choledochotomy. Methods: One hundred and sixty-eight elderly patients with gallbladder and choledocholithiasis were divided into a laparoscopy group(n=75, receiving laparoscopic cholecystectomy and choledochotomy)and an open abdominal group(n=93, undergoing traditional open cholecystectomy and common bile duct exploration). The surgical incision length, operation time, intraoperative blood loss, anal exhaust time, hospital stay and postoperative complications were compared between the two groups. Results: The surgical incision length, operation time, intraoperative blood loss, anal exhaust time, hospital stay were lower in the laparoscopic group than in the open abdominal group(P<0.05). The incidence of postoperative complications was lower in the laparoscopic group than in the open group(P<0.05). Conclusions Laparoscopic cholecystectomy and choledochotomy can obviously improve the clinical efficacy and reduce complications in elderly patients with gallbladder and choledocholithiasis, and it is worthy of clinical application.

Objective: To understand effects of timing and duration of separation experiences from parents on emotion regulation of left-behind preschoolers,and to provide the reference for phychological instruction and intervention among the special groups of children. Methods: Children’s emotion regulation strategy and the relevant information among 1 373 left-behind preschool children from Funan county in Fuyang.Qianshan county in Anqing,Changfeng county and Fexi county inFeixi were investigated. Results: Children with left-behind experiences younger than 18 months old tend to use less cognitive restructuring (P=0.03) and alternative action strategies (P=0.00) than non-left behind children. Children separated from father less than 47 months (median) and 36 months (median) from mother tend to use less cognitive restructuring (P=0.00) and alternative action strategies (P=0.00) than non-left behind children. Conclusion Separation experiences from parents younger than 18 months old exert severe damage on children’s emotional regulation. With the duration of separation increases, children show resilience of emotion regulation, which might be a protective factor for negative emotion due to parent-child separation.

Objective To know the anxiety status of preschool children in rural areas and to explore its influential factors. Methods Subjects of 1363 rural preschool children aged 3-6 years were selected from Anhui Province( Changfeng and Feixi county of Hefei city; Qianshan county of Anqing city; Funan county of Fuyang city) . The primary caregivers of preschool children were investigated by Chinese version of Spence Preschool Anxiety Scale (SPAS),Devereux Early Childhood Assessment for Preschoolers (DECA-P2) ,Neglect Norms for Children Aged 3 to 6 Years in Rural Areas of China,Adaptation Partnership Growth Affection Resolve (APGAR),Self-rating anxiety scale (SAS) and Self-rating depression (SDS). Result-s Among the 1363 preschool children,the positive rate of anxiety was 14. 7％. Physical injury fears score was highest (1. 24±0. 84),followed by social phobia anxiety (0. 86±0. 75) and separation anxiety (0. 85± 0. 74) . Except for obsessive compulsive disorder,the other four types of anxiety and total anxiety scores were negatively correlated with the total protective factors of mental resilience (r=-0. 054- -0. 070,P<0. 05). Besides,all anxiety scores were positively correlated with behavioral problems(r=0. 121-0. 237,P<0. 05)and neglect degree (r=0. 157-0. 269,P<0. 05). The results of multiple linear regression analysis of different anxiety types showed that gender,family type,family income status,parental rearing pattern was consistent or inconsistent,children' s neglect degree,behavior problem and anxiety/depression status of primary caregivers were the main factors affecting the anxiety level of preschool children in rural areas. Conclusions In order to reduce or avoid anxiety of preschool children,it is necessary to establish a good family environment,im-prove parents' upbringing level,and take effective education intervention.