Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Objective:To investigate the clinical features, diagnostic methods, treatment modalities, and prognosis of parathyroid multiglandular disease (PTMGD) in primary hyperparathyroidism (PHPT) .Methods:The clinical data of 29 patients with PTMGD who underwent surgery at the General Surgery Department of the Tianjin Medical University General Hospital from Dec. 2015 to Jul. 2023 were retrospectively analyzed, including the patients' preoperative and postoperative blood calcium and parathyroid hormone, the main clinical manifestations, the involvement of other systems, the main types of pathology, the accuracy of the various examinations, and the postoperative prognosis, etc., and were compared with the 291 patients who had undergone surgery for single-glandular lesion patients were compared. SPSS25 was used to analyze the data.Results:The age of onset of PTMGD was 52.7±1.9 years compared to 56.6±0.7 years in patients with monoglandular disease, P=0.047. Tumor diameter of PTMGD was (2.05±0.1) cm and (2.34±0.6) cm of monoglandular disease, P=0.006. The preoperative blood calcium was (2.56±0.59) mmol/L in PTMGD and (2.70±0.58) mmol/L in monoglandular disease, P=0.045. Preoperative parathyroid hormone (PTH), blood calcium, and Win values were positively correlated with maximum tumor diameter in patients with PTMGD (R-values of 0.362, 0.223, and 0.352, respectively) .Neck ultrasound, neck-enhanced CT and parathyroid nuclear imaging were used to localize and diagnose the diseased parathyroid glands in this group of cases.The accuracy rates were (14/25) 56%, (10/19) 53% and (11/24) 46% in patients with PTMGD, while in patients with monoglandular disease, the accuracy rates were (233/250) 89%, (131/152) 96% and (223/232) 86%. PTMGD accuracy rate was less than that of monoglandular disease,and was statistically significant ( P-value was less than 0.001 in all cases) .The accuracy of the combined localization diagnosis of the three tests in patients with PTMGD was then improved to (13/18) 72%. The pathology of PTMGD was predominantly parathyroid hyperplasia, 45/72 (63%), compared to that of monoadenopathy 18/291 (6%), P<0.001. Parathyroid adenomas predominated in patients with monoadenopathy compared to that of PTMGD, 237/291 (82%) vs. 24/72 (33%), and the proportion of parathyroid adenomas in patients with monoadenopathy was higher than that in patients with PTMGD, P<0.001. 23 patients with PTMGD were followed up, of whom 9 showed mild elevation of parathyroid hormone postoperatively, and 1 patient showed signs of hypoparathyroidism. Conclusion:The low age of onset of multiglandular lesions in primary hyperparathyroidism, mild biochemical tests, and the difficulty of accurately locating all lesions preoperatively warrant adequate preoperative evaluation to promptly identify patients with familial multiple endocrine adenomas, as well as intraoperative bilateral parathyroid exploration in patients with suspected multiglandular lesions.

Objective:To explore the postprandial plasma low-density lipoprotein cholesterol (LDL-C) changes by various detection methods.Methods:A total of 85 subjects admitted to the Second Xiangya Hospital of Central South University from November 2017 to May 2019 were included. Serum samples were collected from fasting and the 2 nd hour and the 4 th hour after breakfast. Serum lipid levels were measured with enzymatic assays and nuclear magnetic resonance spectroscopy (NMRS), and proprotein invertase subtilisin/kexin type 9 (PCSK9) levels were measured with enzyme-linked immunosorbent assays. The differences of blood lipid components at different time points were compared by Friedman two-way rank analysis of variance and Wilcoxon signed rank test, and the correlation between PCSK9 level and lipoprotein particles was analyzed by Spearman correlation. Results:Measured by enzymatic assays, compared with the fasting state, LDL-C decreased at the 2 nd hour and the 4 th hour after the meal (2.58[2.09, 3.12], 2.47[1.92, 3.02], 2.37[1.82, 2.80] mmol/L, P<0.001). Measured by NMRS, the concentration of LDL particles (1 086[830, 1 239], 1 083[848, 1 213], 1 061[814, 1 213] nmol/L, P=0.417) did not change significantly, and cholesterol in LDL particles were 2.13 (1.56, 2.54), 2.16 (1.68, 2.50), 2.06 (1.58, 2.50) mmol/L, respectively ( P=0.047)，and postprandial cholesterol in LDL particles in the 2 nd hour and in the 4 th hour did not change significantly compared with fasting ( P>0.05). while the concentration of large LDL particles (185.2[150.6,221.6], 173.0[144.8,220.3], 178.1[144.0,233.6] nmol/L, P=0.001), and the cholesterol level in large LDL particles (0.49[0.39, 0.57], 0.47[0.38, 0.57], 0.46[0.37, 0.58]mmol/L, P<0.001) decreased after the meal. The PCSK9 level also decreased significantly after the meal (299[233, 397], 257[208, 342], 251[215, 340] ng/ml, P<0.001). There was an independent positive correlation between the decrease of PCSK9 levels and the increase of remnant cholesterol detected by MNRS after the meal ( r=0.232, P=0.035). Conclusions:The postprandial LDL-C level measured by NMRS and enzymatic assays is not consistent. The decrease of LDL-C measured by enzymatic assays is not caused by the clearance of LDL particles, but by the redistribution of cholesterol in each LDL subfraction.

Objective:To investigate the characteristics of perceived stress and coping style and their relationship withsymptoms in college students with psychosis-risk syndrome (PRS).Methods:A two-stage screening process was conducted in 3160 college students.Participants were screened with 16-Item Version of the Prodromal Questionnaire (PQ-16).The students with scores of PQ-16 ≥9 were further interviewed with Structured Interview for Prodromal Syndromes (SIPS).Twenty-two individuals who met the criteria of PRS on SIPS were enrolled as the PRS group.The control group (n =22) were randomly selected from the participants with scores of PQ-16 ＜9 who were matched with age,gender and education level.Two groups were assessed with Chinese Perceived Stress Scale (CPSS),Trait Coping Style Questionnaire (TCSQ) and Self-rating depression scale (SDS) to measure subjective stress,coping style and depressive symptoms.Results:PRS group reported higher CPSS total scores [(34 ± 8)vs.(22 ± 8)] and negative coping styles scores [(35 ± 5)vs.(28 ± 7)],and lowerTCSQ positive coping styles scores [(31 ±6)vs.(37 ±4)] than the controls (Ps ＜0.05).The CPSS scores in PRS group were positively correlated with the scores of negative coping style (r =0.57,P ＜0.01),and the SDS scores were positively correlated with CPSS total scores and scores of negative coping style (r--0.86,0.66,Ps ＜ 0.01).There was no significant relation of psychosis-risk symptoms to scores of CPSS and TCSQ (P ＞0.05).Conclusion:It suggests that college students with PRS have a higher perceived stress and tend to use more negative coping style,which may be positively correlated with depression.