Objective:To investigate the expression levels of miRNA-21 (miR-21) and miRNA-330 (miR-330) in serum exosomes of non-small cell lung cancer (NSCLC) patients with brain metastases, and the correlation of the two with the prognosis of patients.Methods:A prospective cohort study was conducted. A total of 125 NSCLC patients who were admitted to the Affiliated People's Hospital of Shandong First Medical University from March 2021 to September 2022 were prospectively selected, and the brain metastasis was determined by CT, contrast-enhanced magnetic resonance imaging of the head, or surgical pathology. The NSCLC patients were divided into the metastatic group (58 cases) and the non-metastatic group (67 cases) according to whether they had brain metastases, and 50 patients with benign lung diseases and 50 healthy subjects who underwent physical examination in the same period were selected as benign group and healthy control group respectively. Serum samples were collected from all subjects (including patients' pre-treatment samples), the exosomes were extracted, and real-time fluorescence quantitative polymerase chain reaction was used to determine the relative expression of miR-21 and miR-330 in exosomes at the transcriptional level, and electrochemiluminescence immunoassay was used to detect the levels of serum tumor markers [neuron-specific enolase (NSE), carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCCA)]. The levels of miR-21 and miR-330 in serum exosomes and serum tumor markers in the 4 groups were compared, and the correlation between miR-21 and miR-330 in serum exosomes of NSCLC patients with brain metastases before treatment and the correlation between miR-21, miR-330 and serum tumor markers were analyzed by Pearson method. Using brain metastases identified by CT, contrast-enhanced magnetic resonance imaging of the head or surgical pathology as the gold standard, the receiver operating characteristic (ROC) curves were drawn to determine the occurrence of brain metastases in NSCLC patients based on the levels of miR-21, miR-330 and their combination in the serum exosomes before treatment. NSCLC patients were divided into the poor prognosis group and the good prognosis group according to whether or not they died of tumor during the follow-up period, and the clinical characteristics and levels of miR-21 and miR-330 in serum exosomes before treatment were compared between the two groups. The independent influencing factors of death due to tumor in NSCLC patients were analyzed by multivariate logistic regression.Results:Among 125 NSCLC patients, 68 (54.4%) were male and 57 (45.6%) were female; the age was (63±5) years old, ranging from 49 to 82 years old; 89 patients (71.2%) were adenocarcinoma and 36 patients (28.8%) were squamous cell carcinoma. The transcriptional level relative expression of miR-21 in serum exosomes of healthy control group, benign group, non-metastatic group and metastatic group increased sequentially, the transcriptional level relative expression of miR-330 decreased sequentially, the protein concentrations of NSE, CEA and SCCA increased sequentially, and the differences between each two groups were statistically significant (all P<0.001). Pearson correlation analysis showed that in the serum exosomes of NSCLC patients with brain metastases before treatment, miR-21 was positively correlated with serum NSE, CEA and SCCA levels ( r values were 0.641, 0.785 and 0.612, respectively; P values were 0.015, 0.011 and 0.019, respectively), miR-330 in the serum exosomes before treatment was negatively correlated with serum NSE, CEA, and SCCA levels ( r values were -0.612, -0.689 and -0.587, respectively; P values were 0.016, 0.021 and 0.013, respectively), and miR-21 was positively correlated with miR-330 in the serum exosomes before treatment ( r = -0.529, P = 0.023). ROC curve analysis showed that the area under the curve of miR-21, miR-330 and their combination in serum exosomes before treatment for determining the occurrence of brain metastases in NSCLC patients were 0.861 (95% CI: 0.792-0.931), 0.894 (95% CI: 0.840-0.947) and 0.906 (95% CI: 0.849-0.963), and the differences were statistically significant (all P < 0.001). The optimal cut-off value of miR-21 relative expression was 1.625, and the corresponding sensitivity and specificity were 77.4% and 71.5%, respectively; the optimal cut-off value of miR-330 was 0.611, and the corresponding sensitivity and specificity were 81.1% and 74.9%, respectively; the sensitivity and specificity when the two were combined to reach the optimal cut-off value were 84.5% and 73.8%, respectively. NSCLC patients were followed up for a median time of 19 months (95% CI: 17-21 months), and 23 cases (18.4%) died due to the tumor during the follow-up period. The proportions of patients with age ≥60 years old, clinical stage Ⅲ-Ⅳ and brain metastases and the relative expression of miR-21 in serum exosomes before treatment in the poor prognosis group were higher than those in the good prognosis group, the relative expression of miR-330 in the serum exosomes before treatment was lower than that in the good prognosis group, and the differences were all statistically significant (all P < 0.05). Multivariate logistic regression analysis showed that the high age (≥60 years old vs. <60 years old, OR = 3.750, 95% CI: 1.191-11.806, P = 0.024), late clinical stage (stage Ⅲ-Ⅳ vs. stage Ⅰ-Ⅱ, OR = 4.667, 95% CI: 1.303-16.716, P = 0.018), brain metastasis (with metastasis vs. non-metastasis, OR = 2.573, 95% CI: 1.008-6.611, P = 0.049), and elevated relative expression of miR-21 in serum exosomes before treatment ( OR = 2.585, 95% CI: 1.198-6.152, P = 0.008) were the independent risk factors for death due to tumor in NSCLC patients, and elevated relative expression of miR-330 in serum exosomes before treatment was an independent protective factor for death due to tumor ( OR = 0.821, 95% CI: 0.715-0.954, P < 0.001). Conclusions:miR-21 level is high and miR-330 level is low in serum exosomes of NSCLC patients with brain metastases before treatment, and there is a negative correlation between them, and they are closely related to various serum tumor markers of NSCLC patients with brain metastases and NSCLC patients' prognosis; the combination of the two may predict the occurrence status of brain metastases in NSCLC.

Objectives:To investigate the clinical value of endoscopy and mucosal histology for digestive tract diseases in infants.Methods:Clinical data of 357 infants who underwent 422 gastrointestinal endoscopies from January 2010 to December 2021 were collected. The indications, endoscopic manifestations, histological features of mucosa and diagnosis were analyzed.Results:A total of 159 gastroscopies and 263 colonoscopies were performed. Diarrhea (185 cases), bloody stool (178 cases) and vomiting (46 cases) were common symptoms. Endoscopy showed manifestations including non-specific inflammatory changes (265 cases), ulcer-like changes (72 cases), and normal mucusa (48 cases). A total of 373 biopsies were performed, including 260 cases of abnormal mucosal histology, 109 cases of normal mucosal histology, and 4 cases of too small biopsy specimens for analysis. Diagnoses were 208 cases of anaphylactic disease, 45 cases of inflammatory bowel disease, 15 cases of variation of structure, 10 cases of intestinal lymphangiectasis, 2 cases of autoimmune enteropathy, and 1 case of celiac disease. There was only 1 colonic perforation complicating endoscopy in terms of endoscopic complication.Conclusions:It is safe and effective to perform endoscopy standardly for digestive tract diseases in infants. Endoscopy with biopsies is a greatly informative test for diagnosis in infants.

Objective:To analyze the clinical characteristics and efficacy of drug treatment in children with inflammatory bowel disease (IBD) at different ages of onset.Methods:The clinical data of 87 children with IBD admitted to Department of Gastroenterology in Children′s Hospital, Capital Institute of Pediatrics from January 2009 to December 2018 were collected. The patients were divided into four groups according to the age of onset: 0 -<2 years old group (36 cases), 2 -<6 years old group (10 cases), 6 -<10 years old group (12 cases) and 10 -<18 years old group (29 cases). The clinical manifestations, laboratory examination, endoscopic findings, pathologic and genetic changes, and treatment were compared among different age groups with chi-square test or Fisher′s exact text.Results:(1) A total of 87 patients were diagnosed with IBD, including 50 Crohn′s disease (CD) (57%), 25 ulcerative colitis (UC) (29%) and 12 unclassified inflammatory bowel disease (IBD-U) (14%). (2) Patients with fever accounted for 78% (28/36) and 8/10 in the 0 -<2 years old group and 2 -<6 years old group, respectively. Patients with abdominal pain and perianal diseases accounted for 6% (2/36) and 47% (17/36) in the 0 -<2 years old group, and their proportions were significantly different among the four groups (χ 2=8.369, 40.317 and 13.130, all P<0.05). (3) Leukocytosis, thrombocytosis and anemia were more common in the 0-<2 years old group, seen in 72% (26/36), 31% (11/36) and 81% (29/36), respectively. There were significant differences in the changes of complete blood count among the four groups (χ 2=21.919, 8.095 and 11.520, all P<0.05). (4) Colonic involvement accounted for 85% (17/20) in the 0 -<2 years old CD patients. While in the CD patients over 6 years old, 61% (14/23) had inflammation of ileum and colon, with a significant difference compared to that in patients under 6 years old (19% (5/27) , χ 2=9.455, P=0.003). Also, the location of bowel inflammation among the four groups were significantly different (χ 2=21.120, P<0.01). (5) Noncaseating granulomas were found in 15 (30%) CD patients, and crypt abscess was found in 11 (44%) UC patients. (6) Among the 24 patients whose genes were analyzed by high throughput sequencing, 12 had pathogenic single gene mutation. (7) There were 25 patients treated with total enteral nutrition. Among the 25 patients treated with thalidomide, 20 (80%) had clinical remission or partial remission. Among the 19 CD patients treated with infliximab (IFX), 14 had clinical remission at the 6 th week of treatment, and the proportion of remission maintenance at the 30 th week of treatment was 12/14. (8) The rate of clinical remission or partial remission was 64% (23/36) in the 0 -<2 years old group, 8/10 in the 2 -<6 years old group, 11/12 in the 6 -<10 years old group, and 83% (24/29) in the 10 -<18 years old group. Conclusions:The proportion of CD was higher than that of UC in this study. Infant onset inflammatory bowel disease was more likely to present with perianal lesions, and was usually associated with leukocytosis, thrombocytosis and anemia, and has high possibility of single gene mutation. IFX may be effective in treating CD.

Objective To evaluate the efficacy of Huangqi injection combined with ribavirin in the treatment of viral myocarditis in children.Methods A total of 89 children with viral myocarditis were randomly divided into treatment group (45 cases) and control group (44 cases) by random number table method.The control group received ribavirin intravenously on the basis of routine treatment,and the treatment group was given Huangqi injection intravenously on the basis of the control group.Both groups were treated continuously for 14 days.The Serum creatine kinase (CK),creatine kinase MB (CK-MB),lactate dehydrogenase (LDH) and aspartate transaminase (AST) were detected by automatic biochemical analyzer.The Serum IL-6,IL-8 and TNF-α were detected by ELISA.The level of CD3+,CD4+,and the ratio of CD4+/CD8+ in peripheral blood was measured by flow cytometry,and the clinical efficacy was evaluated.Results After treatment,the level of serum CK (110.19 ± 12.17 U/L vs.143.81 ± 17.53 U/L,t=10.530),CK-MB (18.43 ± 4.40 U/L vs.30.28 ± 5.21 U/L,t=11.602),LDH (139.57 ± 10.36 U/L vs.162.18 ± 13.22 U/L,t=8.992),AST (24.14 ± 2.19 U/L vs.35.58 ± 3.34 U/L,t=19.150) in the treatment group were significantly lower than those in the control group (P＜0.01).The Serum level of IL-6,IL-8 and TNF-α in the treatment group were significantly lower than those of the control group (t=14.884,17.043 and 14.364,respectively,P＜0.01).The level of CD3+,CD4+,and the ratio of CD4+/CD8+ in peripheral blood of the treatment group was significantly higher than that of the control group (t=6.256,0.219 and 3.895,respectively,P＜0.01).Conclusions The Huangqi injection combined with ribavirin can significantly reduce the degree of myocardial injury and inflammation in children with viral myocarditis,and improve the immune function of children with viral myocarditis.

Objective To explore the diagnosis and treatment of celiac disease in children. Method The clinical data of 2 children with celiac disease were analyzed retrospectively. Results Both of them were 8-year and 2-year-old males. Chronic diarrhea and malnutrition were the main manifestations combined with anemia and hypoalbuminemia in both of them. The examinations showed that stool Sultan III staining, anti-endometrial antibody IgA, and anti-gliadin IgA were all positive. Endoscopic examination revealed mucosal lesions in duodenal and small bowel. Pathology showed small intestinal villus atrophy, infiltration of lymphocytes in crypt epithelium, and infiltration of lymphocytes and plasma cells in the lamina propria. Diarrhea was quickly improved in both of them after the gluten diet was avoided, and weight increased significantly. However, one of them failed to maintain a gluten free diet and diarrhea recurred. Conclusions The celiac disease in children is rare in China. It needs to have early diagnosis and treatment, and should be strictly followed by a lifelong gluten free diet.

Objective To investigate the CT features of hepatic focal nodular hyperplasia (FNH) in children.Methods Thirteen patients with FNH,which was confirmed by postoperative pathology,were enrolled retrospectively.Plain and contrast CT were performed on all patients before operation.The CT imaging features of FNH including size,shape,density,style of contrast were analyzed retrospectively and compared with pathology.Results There were 13 patients with 16 lesions,8 lesions were found in the right lobe,5 lesions in the left lobe and 3 lesions involving both lobes.The tumor size ranged from 5.5 cm to 11.5 cm (media size 7.5 cm) in diameter.Histologically,2 cases were typical type,11 cases were atypical type.The lesions were regular morphology in 12 cases and 1 case with capsule.On plain CT,the lesions were isodensity (n=1) or slightly low-density (n=12).In 2 typical type lesions,there were slit-like,stellate-shaped low density central scars.Arterial phase demonstrated that 12 cases were significantly enhanced and 1 case showed mild enhancement.The central scar was not enhanced.In 12 cases,thickened and torturous arteries were seen.The enhancement was reduced at the portal venous phase in all the lesions,with 10 cases showing slightly higher density,2cases isodensity and 1 case low-density.Two cases showed mild enhancement of the central scar.The enhancement of the solid portion in all lesions decreased at the delay phase,with 12 cases showing isodensity and 1 case slightly low density.Two cases with central scar showed delayed enhancement with slightly higher density.Conclusion The CT features of FNH in children are diversified but distinctive which are related with postoperative pathological findings.Combining with clinical symptoms and CT features can be helpful for the early diagnosis of FNH in children.

Objective To evaluate wall histological abnormalities 2 to 3 cm to the end in high or intermediate anal atresia in order to identify features that explain postoperative bowel dysfunctions.Methods Sixty anal atresia patients treated in the Capital Pediatric Institution between January 2008 and December 2012 were recruited in our study.36 patients were resected the terminal anal segment (3 cm).Compared with those 24 cases who were not.Resected samples were fixed for HE and immuno-histochemical stainings.Clinical data including sacral ratio (SR),age at operation,gender,bowel function were evaluated.Results There was no significant difference in patients' SR value,gender and age at operation between resected group and control group.The median follow-up period was 4.5 years.The rates of voluntary bowel movement,soiling (grade 1,2,3) were similar in both groups,however,the rates of severe constipation in resection group was significantly lower that in control group (3 ％ vs.21％,P ＜ 0.05) In the bowel wall of distal 2 cm anrectal canal,the connective tissue was found to be irregular and abnormally represented.Muscle coat was abnormal in all cases,showing the dysplasia circular and longitudinal layers.The number of enteric nervous system was significant fewer in distal 2cm anrectal canal than that in distal 3 cm(1.6 ±0.9 vs.5.6 ±1.8,t=11.715,P＜0.01).Conclusions Resection of terminal 3 cm at least of the atresia anal canal benefits postoperative bowel defecation function.

Objective:To construct the rat models of orthotopic bladder cancer induced by N-methyl-nitrosourea (MNU),and to evaluate the value of magnetic resonance imaging (MRI)in the noninvasive diagnosis of the bladder cancer model.Methods:Sixty femail SD rats were divided into experiment group (n=45)and control group (n=15).The rats experiment group were induced with MNU (2 mg per rat)by intravesical administration every other week,for 4 times.Meantime,the rats in control group were treated with normal saline (0.2 mL per rat)by intravesical administration.At the end of the 14th week,all rats were examined by MRI and the pathological changes of bladder tissue were detected.Results:In experiment group,43 rats were alived and 2 rats were died at the end of the 14th week;the survial rate was 95.6% and the death rate was 4.4%;the abnormal signals were found in each of 43 rats by MRI which manifested as bladder tumor, and the same results were identified by pathology;the tumor formation rate was 100%.In control group,14 rats were alived and 1 rat was died at the end of the 14th week;the survival rate was 93.3%,and the death rate was 6.7%;there was no abnormal signal in the MRI examination and no bladder cancer in the pathological examination;the tumor formation rate was 0.The tumor formation rates of bladder cancer of the rats in two groups had significant difference (P <0.05),and the death rate had no significant difference (P >0.05).Conclusion:The method to establish the rat models of orthotopic bladder cancer induced by MNU is simple and reliable;the results of MRI are consistent with the pathological results and MRI examination is a reliable diagnostic method concerning this model.

Objective To investigate possible role of 14 -3 -3 in the pathogenesis of asthma inflammation, the expression of 14 -3 -3 protein was observed in lung tissues of asthmatic rats.Methods Expressions of 14 -3 -3 protein was determined by immunohistochemisty method in lung tissues,and the relationship between 14 -3 -3 and asthma inflammation was analyzed.Results The location of positive expression of 14 -3 -3 protein was mainly at cytoplasm,while little at plasmalemma.The positive expression cell mostly was bronchial epithelial cell,others were lymphocytes,alveolar macrophages and vascular endothelial cells.On the other hand,the bronchial smooth muscle and vascular smooth muscle were negative expressed.Moreover,the expression level of 14 -3 -3 protein in asthma group [(0.353 ±0.023)absorbance]was significantly higher than that in the control group[(0.211 ±0.028 )absorbance] (t =10.969,P <0.01).Conclusion The results showed that the 14 -3 -3 protein was overexpressed in asthmatic lung tissue,it may play an important role in asthma inflammation through bronchial epithelial cells,lymphocytes, alveolar macrophages and vascular endothelial cell.

Objective To investigate the CT features of hepatic mesenchymal hamartoma (HMH)in children.Methods Nine patients with HMH confirmed by postoperative pathology were enrolled,including 4 were males and 5 were females.Their age ranged from 3 days to 9 years 5 months (the median age was 7 months).All patients admitted due to palpable abdominal mass without jaundice.All patients were examed by contrast-enhanced CT before the operation.Results All the 9 cases showed solitary hepatic mass,among which 6 were in the right lobe,2 were in the left lobe and 1 involved in both lobes.The tumor size ranged from 7.0 to 22.5 cm (mean size was 13.5 cm) in diameter.The CT manifestations of HMH was related to the proportion and distribution of component in the masses.The masses were cystic (n =1),cystic-solid mixed (n =6) and solid (n =3).After contrast administration,the solid component and the septa of the mass showed enhancement while cystic component was not enhanced.Calcification was seen inside the tumor in one case.Conclusions The CT features of HMH in children are multitudinous which are related to postoperative pathological findings.With the clinical history,it is easy to distinguish HMH from the other hepatic tumors.