ObjectiveTo solve the problems of delayed growth and development and insufficient spawning of experimental Zebrafish, so as to improve the reproductive efficiency and service life of experimental Zebrafish. MethodsThe zebrafish at the age of 2 months after fertilization were divided into two groups. The experimental group was fed with dry commercial diets specifically designed for ornamental fish or frozen adult brine shrimp, while control group was fed with live laval brine shrimp. Within a period of 70 days, the growth performance of the zebrafish was evaluated by measuring body length and weight, and the reproductive performance was assessed by measuring the fecundity and spawning rate. Zebrafish with apparent goiter disease were fed with dry commercial diets, and the inhibitory effect of the pellets on this disease was evaluated by measuring the diameter of the thyroid enlargement lesion. The three feeding methods were combined, and the feeding plan was optimized. The actual effects of the plan on zebrafish rearing were validated through reproductive performance tests. ResultsStarting from 60 days post-fertilization (dpf) until 111 dpf, the body length and weight of the dry commercial diets feed group gradually surpassed those of control group (all P<0.000 1). From 60 dpf to 96 dpf, the growth trend in body length of the adult brine shrimp group was similar to that of control group, but the female fish in the adult brine shrimp group had significantly higher body weight than the female fish in control group at 75-82 dpf (P<0.000 1). Compared to control group, there was a significant difference in body color between males and females in the adult brine shrimp group, and at 75 dpf, gender could be accurately distinguished by body color differences. Furthermore, the spawning rate of the zebrafish in the adult brine shrimp group at 3 months of age was significantly higher than that of control group (94.44% vs. 27.78%, P<0.05). Additionally, after feeding with the dry commercial diets for 130 days, all thyroid enlargement lesions in the experimental zebrafish disappeared. Based on the above results, the three feeding methods were combined and the feeding plan for zebrafish older than 2 months of age was optimized as follows: feed live brine shrimp in the morning, and alternate between dry commercial diets and adult brine shrimp in the afternoon. This feeding plan lasts until the age of 12 months. The spawning rate of Zebrafish can maintan 70%, and the spawning amount can reach (233.6±3.95) eggs. The fertilization rate and hatching rate were 97.47% and 90.24%, respectively, both significantly higher than those of control group (P<0.001, or P=0.01). ConclusionCompared to live brine shrimp feed, the dry commercial diets feed significantly improves the growth performance of zebrafish and has a therapeutic effect on thyroid enlargement disease. On the other hand, adult brine shrimp feed significantly enhances the early reproductive performance of zebrafish. The optimized feeding plan successfully improves the spawning efficiency of laboratory zebrafish, prolonging their reproductive lifespan and better supporting relevant scientific research.

OBJECTIVES: With the increase in aging population in China, elderly Crohn's disease (CD) patients need to receive more attention. This study aims to explore the clinical characteristics and disease process of elderly onset CD (EOCD) patients in a single center. METHODS: From January 2002 to January 2022, a total of 221 patients with CD from the Seventh Medical Center of Chinese PLA General Hospital were enrolled. According to the Montreal CD classification standard, the patients were further divided into 4 groups: an EOCD group (≥60 years old, n=25), a middle age onset CD (MOCD) group (40-59 years old, n=46), a young onset CD (YOCD) group (17-40 years old, n=131), and a childhood onset CD (COCD) group (6-16 years old, n=19). We compared the clinical characteristics and long-term prognosis among them. RESULTS: Females were predominant in the EOCD group (15/25, 60%). The number of people without smoking in the EOCD group (80%) was lower than that in COCD group (100%), higher than that in the YOCD group (70.2%) and the MOCD group (69.6%) (all P<0.05). Patients with perianal diseases at diagnosis were rare in the EOCD group (0%), lower than that in the COCD group (21.1%) and the YOVD group (19.8%) (all P<0.05). Stenosis was the most common disease behavior in the EOCD group (63.0%), significantly higher than that in the COCD group (15.8%), the YOCD group (36.6%) and the MOCD group (43.5%) (all P<0.05). The EOCD group was easier to be misdiagnosed as tumor (24%), higher than that in the COCD group (0%), the YOCD group (6.9%) and the MOCD group (19.6%) (all P<0.05). The EOCD group was prone to comorbidities (52%), and 20% of them were complicated with multiple comorbidities (P<0.05). During the follow-up, the all-cause mortality of EOCD was 12%, and the CD-related mortality was 8%, which was significantly higher than the other groups (all P<0.05). The use of immunosuppressants in the EOCD group (4.8%) was lower than that in the COCD group (12.8%), the YOCD group (16.8%) and the MOCD group (16.1%), but there was no statistical significance among the 4 groups (P=0.467). In addition, there was no significant difference in the rate of intestinal resection among the 4 groups (P=0.062). CONCLUSIONS In EOCD patients, females were predominant, smoking was less common, and they were prone to comorbidity. At the initial stage of diagnosis, it is easy to be misdiagnosed as tumor, and the disease behavior mainly showed stricture type, less complicated with perianal diseases. During the follow-up, all-cause mortality and CD-related mortality of EOCD patients were significantly higher than those of the non-elderly onset CD patients.
Female ; Middle Aged ; Humans ; Aged ; Child ; Adult ; Adolescent ; Young Adult ; Crohn Disease/epidemiology* ; Prognosis ; Constriction, Pathologic ; Aging ; Hospitals, General

Objective:To prospectively follow up the patients with ileocecal inflammatory lesions, to explore the characteristics of Crohn′s disease(CD) at early stage, and to provide references for early diagnosis of CD.Methods:From January 2013 to December 2018, at Department of Gastroenterology, The Seventh Medical Center of PLA General Hospital, 232 patients with unexplained ileocecal inflammatory lesions under colonoscopy examination were enrolled, which were followed up for more than one year. Chi-square test and Fisher exact probability text were used to compare the patients with early CD, with non-specific enteritis and intestinal tuberculosis in abdominal symptoms (abdominal pain, diarrhea, abdominal distension, constipation, hematochezia, changes in bowel habits), accompanying symptoms (oral ulcer, arthralgia), the proportion of patients with elevated erythrocyte sedimentation rate (ESR) or elevated C-reactive protein (CRP) level, serum antineutrophilic cytoplasmic antibody (ANCA), anti-saccharomyces cerevisiae antibody (ASCA), tuberculosis infection of T cells spot test, positive rate of fecal occult blood, lesion size, morphology, involvement site under endoscopy and histopathological results. Multivariate binary logistic regression was used to analyze the related factors of early CD.Results:Among 232 patients, 155 were males and 77 were females, and the age of first diagnosis was (43.9±13.8) years old. The follow-up period (range) was 27 months (12 to 79 months). Twenty-nine cases (12.5%) were diagnosed as early CD, 45 cases (19.4%) were intestinal tuberculosis, 105 cases (45.3%) were non-specific enteritis, and 53 cases (22.8%) as undetermined. All of 29 patients with early CD had abdominal symptoms, which accounted for 16.9% (29/172) of 172 patients with ileoceccal inflammatory lesion as well as abdominal symptoms. In early CD patients, the proportions of patients with abdominal pain, elevated CRP level and ESR level, positive rate of ASCA, positive rate of tuberculosis infection T cells and percentage of patients with thickened intestinal wall were all higher than those in patients with non-specific enteritis (62.1%, 18/29 vs. 33.3%, 35/105; 13.8%, 4/29 vs. 0; 13.8%, 4/29 vs. 1.0%, 1/105; 24.1%, 7/29 vs. 1.0%, 1/105; 20.7%, 6/29 vs. 3.8%, 4/105; 95.7%, 22/23 vs. 0), and the proportion of patients without abdominal symptoms was lower than that of patients with non-specific enteritis (0 vs. 31.4%, 33/105). And the differences were statistically significant ( χ2=6.692, Fisher exact probability text, χ2=7.162, χ2=17.826, χ2=7.497, Fisher exact probability text, and Fisher exact probability text, all P<0.05). Early CD patients were more likely to have multiple lesion sites (55.2%, 16/29), and mainly deep ulcers (55.2%, 16/29) and ulcers with a long diameter of 5 to 10 mm (39.3%, 11/28). The lesions of non-specific enteritis were mostly confined to the end of ileum (75.2%, 79/105), which were mainly superficial ulcers (41.0%, 43/105) and ulcers with a long diameter less than 5 mm (69.0%, 49/71). The proportion of patients without abdominal symptoms and the positive rate of tuberculosis infection of T cells spot test of early CD patients were both lower than those of intestinal tuberculosis group (0 vs. 15.6%, 7/45 and 20.7%, 6/29 vs. 68.9%, 31/45). The positive rate of ASCA and the proportion of patients with thickened intestinal wall were higher than those of intestinal tuberculosis group (24.1%, 7/29 vs. 0 and 95.7%, 22/23 vs. 11/19), and the differences were statistically significant (Fisher exact probability text, χ2=13.713, Fisher exact probability text and χ2=6.710, all P<0.05). The results of multivariate binary logistic regression analysis showed that abdominal pain and positive ASCA were independent risk factors for early CD (odds ratio ( OR)=2.855, 95% confidence interval ( CI) 1.014 to 8.037, P=0.047; OR=10.033, 95% CI 2.274 to 44.250, P=0.002). Conclusions:Prospective follow-up for more than one year in patients with unexplained ileocecal inflammatory lesions can effectively identify and diagnose early CD. Ileocecal inflammatory lesions with abdominal symptoms are one of the early manifestations of CD. Abdominal pain and positive serum ASCA at the initial diagnosis are independent risk factors for early diagnosis of CD.

Melanoma is a highly invasive and lethal skin malignant tumor derived from melanocytes. It has the characteristics of high early metastasis and high mortality. In recent years, with the in-depth study of the pathogenesis of melanoma, it has been found that epigenetic modification, especially DNA methylation, is considered to be a universal intrinsic feature of melanoma development and evolution. This article reviews the research progress of abnormal DNA methylation genes in melanoma in detail, and summarizes the biomarker effect of DNA methylation genes, suggesting that the detection of abnormal DNA methylation genes in melanoma patients is hopeful as an early screening index and diagnostic marker for melanoma patients.

Melanoma is a highly invasive and lethal skin malignant tumor derived from melanocytes. It has the characteristics of high early metastasis and high mortality. In recent years, with the in-depth study of the pathogenesis of melanoma, it has been found that epigenetic modification, especially DNA methylation, is considered to be a universal intrinsic feature of melanoma development and evolution. This article reviews the research progress of abnormal DNA methylation genes in melanoma in detail, and summarizes the biomarker effect of DNA methylation genes, suggesting that the detection of abnormal DNA methylation genes in melanoma patients is hopeful as an early screening index and diagnostic marker for melanoma patients.

Objective    To investigate the clinical feasibility and safety of uniportal video-assisted thoracoscopic surgery (VATS) without chest tube in enhanced recovery thoracic surgery. Method    The clinical data of patients with pulmonary bulla, pulmonary nodules and mediastinal tumors who underwent uniportal VATS in Department of Thoracic Surgery in the Affiliated Hospital of Inner Mongolia Medical University between January 2015 to May 2018 were retrospectively analyzed. A total of 78 patients did not receive closed thoracic drainage tube (a tube-free group), including 30 males and 48 females aged 32.5±8.3 years, 92 patients closed thoracic drainage tube after operation (a control group), including 38 males and 54 females aged 31.4±13.6 years. The surgery-related indicators, postoperative complications and visual analogue score (VAS) were compared between the two groups. Results    The time of early ambulation and hospital stay after operation in the tube-free group (1.0±0.3 d, 3.3±0.7 d) were significantly shorter than those in the control group (1.8±0.6 d, 5.2±0.8 d) (P=0.000, P=0.000). The VAS pain scores on the first, second and third day after operation in the tube-free group (4.5±1.8, 3.6±2.4, 2.5±1.4) were also significantly lower than those in the control group (6.8±2.2, 5.7±2.9, 3.9±1.2) (P=0.000, P=0.000, P=0.000). Operation time and intraoperative blood loss in the tube-free group (55.3±12.2 min, 21.5±5.1 mL) and the control group (57.1±6.5 min, 22.2±3.5 mL) were not statistically different (P=0.220, P=0.146). There was no pulmonary infection in both groups, and the wound healing rate was 100.0%. There was no significant difference in pneumothorax, pleural effusion, arrhythmia and re-insertion of chest drain between the tube-free group (5 patients, 8 patients, 1 patient, 3 patients) and the control group (1 patient, 4 patients, 2 patients, 1 patient, P=0.145, P=0.134, P=0.885, P=0.499). Conclusion    In strictly screened patients undergoing uniportal thoracoscopic surgery, no thoracic closed drainage tube can relieve postoperative pain, promote early ambulation activities and enhanced recovery of patients.

Objective    To explore the safety and feasibility of spontaneous breathing anesthesia combined with tubeless uniportal thoracoscopy in pulmonary bullae surgery. Methods    Totally 112 patients with pulmonary bullae in the Affiliated Hospital of Inner Mongolia Medical University from March 2015 to May 2017 were enrolled. According to the random number chosen by computer, the patients were randomly divided into two groups: a tubeless group (spontaneous breathing anesthesia combined with tubeless uniportal thoracoscopy) and a control group (uniportal thoracoscopy by general anesthesia with tracheal intubation) . There were 49 males and 7 females with an average age of 25.5±6.5 years in the tubeless group, and 50 males and 6 females with an average age of 23.5±4.5 years in the control group. The difference of the lowest intraoperative arterial oxygen saturation (SaO2), SaO2 at postoperative one hour, operation time, postoperative awakening time, hospital stay, hospitalization cost and postoperative pain score were analyzed. Results    There was no significant difference between the two groups in the operation time, the lowest SaO2, SaO2 at one hour after the operation and the partial pressure of carbon dioxide (PaCO2). The awakening time and duration of postoperative hospital stay in the tubeless group was shorter than those in the control group (P=0.000). The cost of hospitalization in the tubeless group was less than that in the control group (P=0.000). The discomfort caused by urinary tract and visual analogue score (VAS) in the tubeless group were better than those in the control group. Conclusion    It is safe and feasible to use spontaneous breathing anesthesia combined with tubeless uniportal thoracoscopy in pulmonary  bullae resection.

To compare the changes of serum osteocalcin (OC), interleukin-18 (IL-18) in patients with different glucose tolerance and to explore their relationship to carotid atherosclerosis in type 2 diabetes mellitus (T2DM). Methods: According to the result of oral glucose tolerance test (OGTT), 140 research subjects were divided into 3 groups: Normal control group, n=50, Pre-diabetes group, n=30 and T2DM group, n=60 which included in 2 subgroups:Normal carotid intima-media thickness (IMT) subgroup, n=26 and Carotid IMT thickening subgroup, n=34. Carotid IMT, serum OC, IL-18 and glycosylated hemoglobin (HbA1c), fasting glucose, OGTT 2-hour blood glucose (2hPG), TC, TG, LDL-C, HDL-C, body mass index (BMI), insulin resistance index (HOMA-IR) and insulin cell function index (HOMA-β) were measured in all subjects. Pearson correlation and multi liner regression model were conducted to analyze the relevant parameters. Results: From Normal control to Pre-diabetes to T2DM groups, serum OC was decreased and IL-18 was increased gradually, all P<0.05. OC was negatively related to HbA1c (r=-0.426), fasting glucose (r=-0.582), 2hPG (r=-0.489), HOMA-IR (r=-0.456), TC (r=-0.451) and carotid IMT (r=-0.559), all P<0.05; while positively related to HOMA-β (r=0.439), P<0.05. IL-18 was positively related to BMI (r=0.395), HbA1c (r=0.693), fasting glucose (r=0.880), 2hPG (r=0.715), HOMA-IR (r=0.667), TC (r=0.734), TG (r=0.326), LDL-C (r=0.471) and carotid IMT (r=0.857), all P<0.05; while negatively related to HOMA-β (r=-0.678), P<0.05. In T2DM group, carotid IMT was positively related to IL-18 (r=0.817), fasting glucose (r=0.415), HOMA-IR (r=0.356), TC (r=0.396) and TG (r=0.362), all P<0.05; while negatively related to OC (r=-0.588), P<0.05. Multi liner regression analysis indicated that IL-18, OC, TC and fasting glucose were the independent impact factors for carotid IMT (regression coefficients were 0.013, -0.011, 0.044 and 0.044 respectively), P<0.05. Conclusion: Serum OC and IL-18 had been involved in glucolipid metabolism and closely related to the occurrence and development of carotid atherosclerosis in T2DM patients.

Objective To detect the expression of BRAF V600E mutant protein in cutaneous malignant melanoma (CMM),and to evaluate the sensitivity and specificity of immunohistochemistry (IHC) in detecting BRAF V600E mutation.Methods IHC with an anti-BRAF V600E monoclonal antibody was performed to detect the expression of BRAF V600E mutant protein in paraffin-embedded tissue sections from 103 patients with CMM and 40 patients with nevus.Statistical analysis was carried out with SPSS software version 17.0,and the expression rate of BRAF V600E mutant protein was compared by chi-square test.Results The expression rate of BRAF V600E mutant protein in the CMM patients was 20.4％ (21/103),significantly higher than that in the nevus patients (5.0％ (2/40),x2 =5.06,P ＜ 0.05).Significant differences were observed in the expression rate of BRAF V600E mutant protein between CMM patients of different age groups (29.8％ (14/47) in patients aged ＜ 60 years vs.12.5％ (7/56) in those aged ≥ 60 years,P ＜ 0.05) and nationality (30.2％ (13/43) for Uygur nationality vs.13.3％ (8/60) for Han nationality,P ＜ 0.05),as well as among CMM lesions from different anatomical sites (13.6％ (6/42) in acral sites vs.11.8％ (4/29) in mucous membrane vs.45.8％ (11/32) in non-acral sites,P ＜ 0.05) and of different Clark levels (8.6％ (4/42) for grade Ⅰ-Ⅲ vs.12.4％ (17/61) for grade Ⅳ-Ⅴ,P＜ 0.05),but not between male and female CMM patients or between CMM patients with lymph node metastasis and those without (both P ＞ 0.05).IHC with the anti-BRAF V600E antibody showed a sensitivity of 100％ (15/15) and a specificity of 98.5％ (65/66) in detecting BRAF V600E mutation.Conclusions The expression of BRAF V600E mutant protein is up-regulated in CMM lesions,and CMM patients of Uygur nationality seems to have a higher expression rate than those of Han nationality.IHC appears to be an accurate and rapid method to detect V600E BRAF mutation.

Objective To assess the relationship between BRAF gene mutations and clinical phenotype of malignant melanoma.Methods Tissue specimens were collected from the lesions of 80 patients with malignant melanoma,and from the normal skin of 30 patients with trauma in the Department of Plastic Surgery or General Surgery,and subjected to paraffin embedding and DNA extraction.PCR was performed to amplify the exon 11 and 15 of BRAF gene followed by DNA sequencing.Chi-square test and Fisher's exact test were carried out to assess the relationship between BRAF gene mutations and clinical phenotypes of malignant melanoma.Results BRAF gene mutations were found in 19 (23.8％) of the 80 malignant melanoma specimens.Among the 19 mutationpositive specimens,17 (88.2％) carried mutations in exon 15 of BRAF gene with V600E as the most frequent (88.2％,15/17) mutation type,and 2 (10.5％) carried mutations in exon 11.No mutation was found in any of the normal skin tissue specimens.The average age at onset was 57.5 years in these patients.The frequency of BRAF gene mutation was significantly higher in patients younger than 60 years than in those older than 60 years (37.1％ vs.13.3％,x2=6.613,P ＜ 0.05).A significant difference was observed in the frequency of BRAF gene mutation among tissue specimens of mueosal,acral and non-aeral malignant melanoma (18.2％ (4/21) vs.14.7％(5/34) vs.41.7％ (10/24),x2=6.167,P ＜ 0.05).There was no significant association between BRAF gene mutation and gender,race or lymph node metastasis (all P ＞ 0.05).Conclusions BRAF gene is a hot spot for mutations in patients with malignant melanoma in Xinjiang Uygur Autonomous Region,with V600E point mutation in exon 15 as the most frequent mutation type.BRAF gene mutations appear to be closely correlated with the age at onset of and lesional sites in,but uncorrelated with gender and race of or lymph node metastasis in,patients with malignant melanoma.