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.

Although the artificial intelligence diagnosis technology of traditional Chinese medicine(TCM)is constantly developing,the existing intelligent syndrome differentiation models can only output the diagnosis results and cannot display the reasoning process of TCM diagnosis.They do not have the interpretability of TCM diagnostic knowledge and are difficult to truly reflect the process of TCM's syndrome differentiation.This can be a long-standing problem that has puzzled the academic community.This article elaborates on the development process and technical characteristics of intelligent diagnosis in traditional Chinese medicine,explores multiple key technologies and bottlenecks in current research in reasoning of TCM's Syndrome Differentiation,and factors affecting diagnosis.A novel intelligent diagnosis model architecture that can infer and explain the process of TCM'ssyndrome differentiation was proposed,and the implementation process of the model and algorithm was explained,and detailed data instance analysis was conducted.This article will provide new methods for the development of intelligent syndrome differentiation of TCM and open up new research ideas for clinical decision-making assistance of TCM.

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.

Objective:To explore the effect of the interaction between B7H3 and fibronectin (FN) on the apoptosis of human chronic myeloid leukemia K562 cells.Methods:The expression of B7H3 molecules in K562 cells was detected using flow cytometry and B7H3 overexpressing cells were constructed. The interaction between B7H3 and FN was detected using the co-immunoprecipitation technology. After adding exogenous FN, cell experiments were performed to detect changes in adhesion and cell apoptosis. The changes in apoptosis-related proteins and PI3K/AKT signaling pathway were detected using Western blot.Results:The expression of B7H3 was low in K562, and the cell line K562 OE (overexpression) -B7H3 and the control cell line K562 NC (negative control) -B7H3 were obtained after lentivirus transfection. There is an interaction between B7H3 and FN ( P=0.036) , and this interaction promoted cell adhesion ( P<0.05) , inhibited cell apoptosis ( P<0.05) , and activated the PI3K/AKT signaling pathway ( P<0.05) . Conclusion:B7H3 interacts with FN to promote cell adhesion and may inhibit K562 cell apoptosis by activating the PI3K/AKT signaling pathway.

Objective:To investigate the effects of B7-H3 molecule on clear cell renal cell carcinoma (786-O) metastasis.Methods:Lentiviral transfection method was used to construct 786-O cells stably expressing low level of B7-H3 (shB7-H3 group) and a negative control cell line (shNC group). RT-qPCR, flow cytometry and Western blot were used to assess the efficiency of lentiviral transfection. CCK-8 method was used to detect the proliferation of 786-O cells in the two groups. Flow cytometry was performed to detect the changes in cell cycle. Cell scratch test and Transwell assay were used to detect the differences in cell migration and invasion. Western blot was used to detect the expression of marker proteins in the process of epithelial-mesenchymal transition (epithelial-mesenchymal transition, EMT). Changes in the expression of chemokines and their receptors were analyzed by flow cytometry and RT-qPCR. Effects of anti-CCL4 antibody on cell migration and invasion were analyzed by Transwell assay.Results:Flow cytometry showed that 786-O cells highly expressed B7-H3 molecules and the lentiviral transfection method successfully constructed the cell line with lower expression of B7-H3 (786-O-shB7-H3) and control cell line (786-O-shNC). B7-H3 molecule had no significant effect on the proliferation of 786-O cells. No significant difference in cell cycle was found between the two groups. Compared with 786-O-shNC cells, the migration and invasion ability of 786-O-shB7-H3 cells was suppressed. Moreover, the expression of EMT-related marker proteins (fibronectin and N-cadherin) was reduced and the expression of E-cadherin was increased in 786-O-shB7-H3 cells. The expression of CCL4 and its receptor CCR5 in the shB7-H3 group was lower than that in the shNC group. After intervention with anti-CCL4 antibody, the migration and invasion ability of 786-O-shNC cells was reduced, while that of 786-O-shB7-H3 cells had no significant change.Conclusions:Knocking down the expression of B7-H3 molecule had no significant effect on the proliferation of 786-O cells, but could affect the EMT process of 786-O cells and reduce tumor migration and invasion ability, thereby inhibiting tumor progression.

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 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 evaluate the outcome of patients with alcoholic liver disease (ALD) after orthotopic liver transplantation (OLT) and to study the prognostic factors.Methods The data of 17 patients who underwent OLT for ALD from January 2010 to March 2013 were analyzed retrospectively.The data on age,gender,history of gastrointestinal bleeding,history of splenectomy,Child-Pugh score,Maddrey' s discriminant function and MELD score were evaluated using the Kaplan-Meier method for univariate analysis.The log-rank test was applied to compare the survival rates.Results The overall survival rate at 100 weeks in patients less than 55 years old was 90％ (9/10),while that in patients more than 55 years old was 28.57％ (2/7).There was a significant difference between the two groups (P ＜ 0.05).There were no statistically significant differences between female and male patients,between patients with or without a history of gastrointestinal bleeding or splenectomy (P ＞ 0.05).There was no significant difference on survival rates after liver transplantation between patients with Child-Pugh B and Child-Pugh C,patients with Maddrey' s scores ＜ 70 and ≥70,and patients with a MELD score ＜ 30 and ≥ 30 (P ＞ 0.05).Conclusions Better survival rates were observed in ALD patients who were ＜ 55 years old after liver transplantation.The Child-Pugh score,Maddrey' s discriminant function and MELD score were of no prognostic significance.These findings still need to be verified by prospective large-scale studies.

Objective To explore the change of CD4+CD25+Foxp3+ regulatory T (Treg) cells during aging and the relation with lung tumor. Methods The Lewis lung cancer model was set up in C57BL/6 female mice, and the 36 mice were divided into young health group, middle-aged health group, elderly health group, young tumor group, middle-aged tumor group and elderly tumor group. The percentages of CD4+CD25+Foxp3+ Treg in CD4+ T cells in mice spleen cells were measured by flow cytometry, for reflecting the quantity of CD4+CD25+Foxp3+ Treg cells. And the level of Foxp3 mRNA in splenocyte was tested by real-time PCR method. Results The level of CD4+CD25+Foxp3+/CD4+ T cells and the quantity of Foxp3 mRNA were higher in tumor groups than in healthy groups(both P＜0.05 ). Besides, in the healthy groups, there were statistical differences in the level of CD4+CD25+Foxp3+/CD4+ T cells (F=47.70, P=0.000) and the quantity of Foxp3 mRNA among the different months groups. Accumulation of the CD4+CD25+Foxp3+ Treg cells was accompanied with aging, the elderly mice contained a significantly larger population of CD4+CD25+Foxp3+ Treg cells in their spleen when compared with the younger counterparts, and the highest was the elderly tumor group. So it was with the functional gene Foxp3 mRNA (F=6.56, P=0.009). Conclusions The results suggest a close relationship of the change of CD4+CD25+Foxp3+Treg cells with aging and the genesis and development of lung tumor.