Functional constipation （FC） is a clinically common functional bowel disorder characterized by a protracted course and associations with various chronic disorders and psychological abnormalities. Although not life-threatening， FC significantly impairs patients' quality of life. FC subtypes include slow-transit constipation （STC）， defecatory disorder （DD）， and normal-transit constipation （NTC）. The pathological mechanisms underlying FC have not been fully elucidated， and overall clinical efficacy remains unsatisfactory. Animal models of FC serve as essential tools for the study of disease mechanisms and the development of novel therapeutics. This article systematically reviews the current state of research on the animal models of FC and identifies that rodents， particularly rats and mice， are the most commonly used species. Dogs and pigs are also employed in complex intervention studies due to their physiological similarities to humans， though their use is limited by housing challenges and ethical considerations. Induction methods vary across different FC subtypes. STC models are primarily established with chemical agents such as loperamide or compound diphenoxylate. DD modeling often involves low-fiber diets combined with methylene blue injection or rectal narrowing. NTC modeling mainly relies on low-fiber dietary interventions. In addition， disease-syndrome combination models based on traditional Chinese medicine （TCM） theory have been developed， encompassing excess patterns such as heat accumulation， cold accumulation， and Qi stagnation， as well as deficiency patterns including Qi deficiency， blood deficiency， Yin deficiency， and Yang deficiency. These are achieved through an approach of disease model + syndrome induction， enabling the integration of mechanisms from both Western and TCM perspectives. Models are evaluated from two aspects： disease and syndrome manifestations （e.g.， colonic transit， secretory function， and TCM syndrome indicators such as mental state and body weight） and disease mechanisms （e.g.， enteric nervous system， interstitial cells of Cajal， smooth muscle cells， gut microbiota， and metabolites）. However， current research still faces challenges such as poor consistency in some models， non-specific interference in mechanism interpretation， insufficient studies on NTC， and lack of TCM tongue and pulse diagnosis in evaluation. Future efforts should focus on optimizing model stability and specificity to provide a more reliable experimental basis for investigating the pathological mechanisms of FC and developing therapeutic agents.

Objective:To investigate the correlation between mutations in KRAS, NRAS, BRAF, and PIK3 CA and the clinical characteristics of elderly colorectal cancer(CRC)patients. Methods:Paraffin-embedded tissue samples were obtained from 191 elderly CRC patients who consulted at Huadong Hospital, affiliated to Fudan University, between January 2022 and July 2023.Following deoxyribonucleic acid(DNA)extraction, the amplification refractory mutation system polymerase chain reaction(ARMS-PCR)was employed to detect the mutation profiles of KRAS, NRAS, BRAF, and PIK3 CA.Concurrently, serum samples collected prior to radical resection were analyzed for carcinoembryonic antigen(CEA), carbohydrate antigen 19-9(CA19-9), and carbohydrate antigen 72-4(CA72-4)using electrochemical luminescence.A comparative analysis of the clinical characteristics and preoperative serological tumor marker concentrations among patients with different gene mutations was conducted to elucidate their correlation. Results:A total of 191 elderly CRC patients were enrolled in the study, with ages ranging from 60 to 94 years(mean age 72.1±7.8 years), including 112 males.The mutation rate of KRAS, NRAS, BRAF, and PIK3 CA, as determined by combined detection, was found to be 49.21%(94/191)among elderly CRC patients. KRAS exhibited the highest mutation rate at 35.08%, with statistically significant differences observed in gender, primary site, degree of differentiation, and neurovascular invasion between patients with and without KRAS mutations( P<0.05 for all comparisons).The BRAF mutation rate was 8.90%, and significant differences in gender, age, primary site, and degree of differentiation were also noted between patients with and without BRAF mutations( P<0.05 for all).The mutation rates for NRAS and PIK3 CA were 2.62% and 5.24%, respectively, with no statistically significant differences in the clinical characteristics of patients across different groups( P>0.05 for all).Additionally, the proportion of patients over the age of 90 in the double mutation group was significantly higher( P<0.01).Significant differences in serum CA19-9 concentrations were observed among the various mutation types( P<0.05). Conclusions:There are notable differences in age, gender, primary site, degree of differentiation, and neurovascular invasion among elderly CRC patients with varying mutation statuses of KRAS, NRAS, BRAF, and PIK3 CA.Patients with double mutations exhibited higher concentrations of CA19-9 in preoperative serum.

Objective:To investigate the correlation between mutations in KRAS, NRAS, BRAF, and PIK3 CA and the clinical characteristics of elderly colorectal cancer(CRC)patients. Methods:Paraffin-embedded tissue samples were obtained from 191 elderly CRC patients who consulted at Huadong Hospital, affiliated to Fudan University, between January 2022 and July 2023.Following deoxyribonucleic acid(DNA)extraction, the amplification refractory mutation system polymerase chain reaction(ARMS-PCR)was employed to detect the mutation profiles of KRAS, NRAS, BRAF, and PIK3 CA.Concurrently, serum samples collected prior to radical resection were analyzed for carcinoembryonic antigen(CEA), carbohydrate antigen 19-9(CA19-9), and carbohydrate antigen 72-4(CA72-4)using electrochemical luminescence.A comparative analysis of the clinical characteristics and preoperative serological tumor marker concentrations among patients with different gene mutations was conducted to elucidate their correlation. Results:A total of 191 elderly CRC patients were enrolled in the study, with ages ranging from 60 to 94 years(mean age 72.1±7.8 years), including 112 males.The mutation rate of KRAS, NRAS, BRAF, and PIK3 CA, as determined by combined detection, was found to be 49.21%(94/191)among elderly CRC patients. KRAS exhibited the highest mutation rate at 35.08%, with statistically significant differences observed in gender, primary site, degree of differentiation, and neurovascular invasion between patients with and without KRAS mutations( P<0.05 for all comparisons).The BRAF mutation rate was 8.90%, and significant differences in gender, age, primary site, and degree of differentiation were also noted between patients with and without BRAF mutations( P<0.05 for all).The mutation rates for NRAS and PIK3 CA were 2.62% and 5.24%, respectively, with no statistically significant differences in the clinical characteristics of patients across different groups( P>0.05 for all).Additionally, the proportion of patients over the age of 90 in the double mutation group was significantly higher( P<0.01).Significant differences in serum CA19-9 concentrations were observed among the various mutation types( P<0.05). Conclusions:There are notable differences in age, gender, primary site, degree of differentiation, and neurovascular invasion among elderly CRC patients with varying mutation statuses of KRAS, NRAS, BRAF, and PIK3 CA.Patients with double mutations exhibited higher concentrations of CA19-9 in preoperative serum.

Objective To investigate the computed tomography (CT) features of solitary nodular invasive mucinous lung adenocarcinoma (IMA) in stage IA and establish its prediction model. Methods We included 53 lesions of 53 patients with stage-IA IMA and 141 control lesions of 141 patients with invasive non-mucinous lung adenocarcinoma (NIMA) that were confirmed by surgical pathology in our hospital from January 2017 to December 2019. Univariable analysis was used to compare the demographics and CT signs of the two groups. Multivariable logistic regression analysis was performed to determine the main factors influencing solitary nodular IMA. A risk score prediction model was constructed based on the regression coefficients of the main influencing factors. A receiver operating characteristic (ROC) curve was used to assess the performance of the model. Results The univariable analysis showed significant differences between the two groups in age, largest nodule diameter, tumor-lung interface, lobulation, spiculation, air-bronchogram or vacuole sign, vessel abnormalities (P < 0.05). The spiculation sign was different between the two groups, which was longer and softer in the IMA group while shorter and harder in the NIMA group. There was no significant difference in sex, nodule shape, or pleural retraction (P > 0.05), but irregular shapes were slightly more frequent in the IMA group. The multivariable logistic regression analysis showed that obscure tumor-lung interface (odds ratio (OR = 20.930, P < 0.05), air-bronchogram or vacuole sign (OR = 7.126, P < 0.05), spiculation sign (OR = 4.207, P < 0.05), and vessel abnormalities (OR = 0.147, P < 0.05) were the main influencing factors. The prediction model based on those factors’ regression coefficients had an area under the ROC curve of 0.829 (P < 0.05). Conclusion Compared with those with NIMA, patients with solitary nodular IMA in stage IA were older and more likely to have the CT features of obscure tumor-lung interface, air-bronchogram or vacuole sign, and longer and softer spiculation. Based on the regression coefficients of tumor-lung interface, air-bronchogram or vacuole sign, spiculation, and vessel abnormalities, the risk score prediction model showed good predictive performance for solitary nodular IMA.

Objective:To investigate the mutation of epidermal growth factor receptor (EGFR), the expression of programmed death ligand 1 (PD-L1), cell proliferation-associated antigen (Ki-67) in elderly patients with non-small cell lung cancer (NSCLC), and their correlation with clinical feature such as gender, histological type and TNM stage.Methods:The tissue samples of 340 elderly NSCLC patients with definite histopathological diagnosis were collected from January 2020 to December 2020 in Huadong Hospital Affiliated to Fudan University, including 195 males and 145 females, age between 68.9±6.0 years. Patients were grouped according to clinical features such as gender, histological type and TNM stage. The expression of EGFR mutation, PD-L1 and Ki-67 were detected by Super-ARMS and immunohistochemistry. The correlation between tnem and clinical features was statistically analyzed, and the correlation between EGFR mutation and PD-L1/Ki-67 expression was further analyzed separately.Results:In elderly NSCLC patients′ tissues, the positive rate of EGFR mutation was 48.53% (165/340). L858R and 19del mutations were the most common types, which were 56.36% (93/165), 30.30% (50/165) respectively. The mutation rate of EGFR was higher in women, lung adenocarcinoma, well-differentiated, and low-stage patients, which were 65.52% (95/145), 53.77% (164/305), 56.75% (143/252), 52.53% (135/257) respectively. In addition, the positive rate of PD-L1 expression was higher in elderly patients with non-adenocarcinoma lung cancer and poorly differentiated adenocarcinoma, which were 37.14% (13/35), 24.53% (13/53) respectively. The negative rate of PD-L1 expression was higher in elderly patients with NSCLC in stage Ⅰ+Ⅱ, no lymph node metastasis and weakly positive Ki-67, which were 89.11% (229/257), 87.63% (248/283), 94.71% (197/208) respectively. Correlation analysis showed that EGFR mutation was negatively correlated with the expression of PD-L1 and Ki-67 (PD-L1: r=-0.22, P<0.001; Ki-67: r=-0.32, P<0.001). Conclusion:There is a negatively correlation between EGFR mutation and the expression of PD-L1 and Ki-67 in elderly NSCLC, suggesting that the combined detection of EGFR mutation and PD-L1 expression could provide the basis for precise targeted therapy for elderly NSCLC patients.

Objective:To investigate the effects of hyperuricemia on the prognosis of IgA nephropathy (IgAN) using propensity score matching (PSM) method.Methods:IgAN patients proven by biopsy were included. PSM was used to match patients. Kaplan-Meier method was used for survival analysis, and Cox regression analysis was used to analyze the effects of hyperuricemia on IgAN prognosis. Primary outcome events were defined as death, or end-stage renal disease (dialysis, transplantation), or a decrease in estimated glomerular filtration rate (eGFR) greater than 40%. Renal outcome was defined as end-stage renal disease (dialysis, transplantation), or a decrease in eGFR greater than 40%.Results:A total of 1 454 IgAN patients were included in this study, including 850 females and 604 males. Uric acid level was (368.26±92.87) μmol/L in the males, and (277.23±92.71) μmol/L in the females. The median follow-up time was 85.00(56.10, 106.33) months. During the follow-up period, a total of 134 patients reached the primary outcome events, including 5 deaths, 24 dialysis patients, 5 kidney transplant patients, and 100 patients with eGFR decreased by more than 40%. After 1∶1 matching, 131 males and 159 females in the hyperuricemia group were successfully matched with 131 males and 159 females in the normal uric acid group, and there was no significant statistical difference in each parameter in baseline between the hyperuricemia group and normal uric acid group after matching. Kaplan-Meier survival analysis showed that either before or after matching, the incidence of primary outcome events in male or female patients with hyperuricemia was higher than those with normal uric acid, but there was no statistically significant difference in incidence of primary outcome events between female hyperuricemia group and female normal uric acid group after matching (Log-rank test, χ2=3.586, P=0.058). Cox proportional hazard regression model showed that, in the pre-match fully adjusted model, the hazard ratio ( HR) of entering primary outcome events was 2.29-fold (95% CI 1.27-4.11, P=0.006) for men with hyperuricemia and 1.85-fold (95% CI 1.01-3.37, P=0.045) for women with hyperuricemia compared with those with normal uric acid. In the post-match fully adjusted model, the HR of entering primary outcome events was 2.41-fold (95% CI 1.18-4.93, P=0.016) for men with hyperuricemia and 1.83-fold (95% CI 0.91-3.67, P=0.091) for women with hyperuricemia compared with those with normal uric acid. In the pre-match fully adjusted model, the HR of entering renal outcome events was 2.68-fold (95% CI 1.47-4.88, P=0.001) for men with hyperuricemia and 1.81-fold (95% CI 0.99-3.33, P=0.056) for women with hyperuricemia compared with those with normal uric acid. In the post-match fully adjusted model, the HR of entering renal outcome events was 2.89-fold (95% CI 1.36-6.15, P=0.006) for men with hyperuricemia and 1.81-fold (95% CI 0.88-3.72, P=0.106) for women with hyperuricemia compared with those with normal uric acid. Conclusion:Hyperuricemia may be associated with IgAN progression, and it has a more significant effect on male IgAN patients.

Objective:To study the correlation between circulating tumor cells (CTC) and the degree of pathological invasion, recurrence and metastasis of urothelial carcinoma, and so to explore the clinical value of CTC detection in bladder cancer.Methods:A total of 142 patients with urothelial carcinoma in Huadong Hospital Affiliated to Fudan University were enrolled as cancer group from July 2016 to January 2018. According to the degree of tumor invasion, cancer group was divided into the non-muscle-invasive group (49 cases) and the muscle-invasive group(93 cases). In addition, 52 patients with benign urinary tract lesions admitted were selected as the benign group and 56 patients with non-urinary tract diseases and non-tumor as the control group. A total of 3.2 ml of venous anticoagulant blood from each subject was collected. CTC was enriched by negative enrichment using the magnetic beads coated with monoclonal antibody Cluster 45 of differentiation (CD45) to capture and remove white blood cells, and identified by chromosome 8 probe(CEP8) fluorescence in situ hybridization (FISH) technique. CD45-/4′,6′-diamidino-2-phenylindole+/CEP8>2(CD45-/DAPI+/CEP8>2) cells were judged as CTC. SPSS22.0 statistical software was used for statistical analysis.Results:≥2 CTCs/3.2 ml in blood was set as cutoff value. CTC positive rates in bladder cancer group, benign group and control group were 70.42%(100/142), 28.85%(15/52) and 8.93%(5/56), respectively, and there was a significant difference (χ 2=70.496, P=0.000). There was a statistically difference ( U=2 863.5, P=0.011) in the mean count of CTC(2 CTCs/3.2 ml vs 4 CTCs/3.2 ml) between the two groups. The proportion of≥5 CTCs/3.2 ml in the muscle-invasive group was 40.86% (38/93), which was significantly higher than that in the non-muscle-invasive group, 18.37% (9/49) (χ 2=7.330, P=0.007). Cystoscope follow-up of 65 patients treated with transurethral resection of the bladder tumor showed that the recurrence and metastasis rate in patients with≥5 CTCs/3.2 ml was as high as 47.62% (10/21), compared with 11.36% (5/44) of patients with<5 CTCs/3.2 ml (χ 2=10.530, P=0.001). Among 59 patients undergoing radical cystectomy, no significant difference was found in tumor diameter >3 cm, positive surgical margins and positive lymph nodes among all groups according to CTC negative or positive and CTC number ( P>0.05). But the recurrence and metastasis rate of patients with ≥5 CTCs/3.2 ml (59.10%) was significantly higher than that of patients with <5 CTCs/3.2 ml (6/30)(χ 2=8.364, P=0.004). Conclusion:The number of CTC increased with the deepening of tumor invasion; Tumor recurrence and metastasis increased significantly in the patients with ≥5/3.2 ml CTCs in blood.

Objective To evaluate the diagnostic value of circulating tumor cells(CTCs) in prostate cancer (Pca) through studying the relationship between CTCs and Gleason scores and pathological TNM stage in Pca patients. Methods A total of 238 patients including 161 Pca patients as cancer group, 35 male patients with benign prostatic diseases as benign group and 42 male with non-prostate disease as control group, who were treated in our hospital from July 2016 to January 2018,were enrolled. Venous blood of every patient was collected and CTCs were enriched and identified by immunocytochemistry CD45 capturing leukocyte and fluorescence in situ hybridization with chromosome 8 (CEP8-FISH). Cells displaying CD45-/DAPI+/CEP8>2 were characterized as CTCs. One-way ANOVA was used to exam the correlations of the number of CTCs with Gleason scores and pathological TNM stage. Results CTCs ≥2 were detected in 74.53％(120/161) of Pca patients and 20.00％(7/35)of benign prostatic diseases patients and 7.14％(3/42)of control group (χ2=79.605,P<0.05). In group Gleason scores 6, the numbers of CTCs were 2.00 ± 2.42, the ratios of CTCs≥5 and tetraploid were 13.33％ (2/15)and 26.67％(4/15) respectively. In 7 scores group, the results were 3.14±2.68,17.72％(14/79) and 34.18％(27/79)respectively;In 8 scores group, the results were 3.57 ± 2.70, 33.33％(7/21)and 42.86％ (9/21)respectively; In 9 scores group, these three results were 4.65±4.41, 43.48％(20/46) and 45.65％(21/46)respectively. The numbers of CTCs in the≤pT2b (20), pT2c(27), pT3a(19), pT3b(16)and≥pT4(12) groups were 2.25±2.45, 3.56±2.79, 4.05±3.47, 4.69±2.12 and 5.17±3.21 respectively. The ratios of CTCs≥5 were 25.00％(5/20), 25.93％(7/27), 26.32％(5/19), 50.00％(8/16) and 58.33％ (7/12)respectively. The proportions of tetraploid were 20.00％(4/20), 25.93％ (7/27), 31.58％(6/19), 50.00％(8/16) and 58.33％(7/12) respectively. There were significant differences between CTC and Gleason scores (F=3.200, P<0.05)and pathological stage (F=2.673, P<0.05). The ratios of CTCs≥5 increased with the increase of Gleason scores (χ2=11.592, P<0.05). Conclusions The detection of CTCs could be used for the differential diagnosis of Pca and benign prostatic disease. There were notable correlations between the numbers of CTCs and Gleason scores and pathological stage in Pca patients, especially between CTCs≥5 and Gleason scores.

Objective: To evaluate the impact of KIT D816 mutation on the salvage therapy in relapsed acute myeloid leukemia (AML) with t(8;21) translocation. Method: The characteristics of the first relapsed AML with t(8;21) translocation from 10 hospitals were retrospectively collected, complete remission (CR₂) rate after one course salvage chemotherapy and the relationship between KIT mutation and CR₂ rate was analyzed. Results: 68 cases were enrolled in this study, and 30 cases (44.1%) achieved CR₂. All patients received KIT mutation detection, and KIT D816 mutation was identified in 26 cases. The KIT D816 positive group had significantly lower CR₂ compared with non-KIT D816 group (23.1% vs 57.1%, χ²=7.559, P=0.006), and patients with longer CR₁ duration achieved significantly higher CR₂ than those with CR₁ duration less than 12 months (74.1% vs 31.9%, χ²=9.192, P=0.002). KIT D816 mutation was tightly related to shorter CR₁ duration. No significant difference of 2 years post relapse survival was observed between KIT D816 mutation and non-KIT D816 mutation group. Conclusion KIT D816 mutation at diagnosis was an adverse factor on the salvage therapy in relapsed AML with t(8;21) translocation, significantly related to shorter CR1 duration, and can be used for prediction of salvage therapy response. KIT D816 mutation could guide the decision-making of salvage therapy in relapsed AML with t(8;21) translocation.

In recent years, researches about liquid biopsies which contain circulating tumor cells (CTC), circulating tumor DNA (ctDNA) and exosomes in the body fluids of patients with solid tumor have caused great concern. Compared with traditional biopsies, liquid biopsies have many advantages as they are real time, noninvasive and comprehensive. With the progress in the detecting techniques of CTC, ctDNA and exosomes, liquid biopsies can be applied to the early screening, diagnosis, treatment, prognostic evaluation of tumor. This review focuses on the recent advances in detection methods and clinical applications of CTC, ctDNA and exosomes.(Chin J Lab Med, 2018, 41: 621-626)