Objective To investigate the changes and mechanism of learning and memory in rats by different doses of benzo (a) pyrene (B(a)P). Methods Forty weaned rats (28 days) were randomly divided into control group (NS), solvent group ( vegetable oil) and three B (a) P dosage groups (the doses were 5,10 and 20 mg / kg body weight respectively ). And all rats were administrated intraperitoneally every other day to one month. The capability of learning and memory in rats were measured by Morris water maze test, and the brain-derived neurotrophic factor ( BDNF) and NMDAR2B content in hippocampus were tested by immunohistochemistry. Results In training of Morris water maze,the average escape latency was extended gradually with increasing dose, and there was a statistically significant difference between high-dose group((62. 78 ±47. 25 )s) and the control group((40.60±38.79)s)(P＜ 0.01). Compared with the control group(11.25 ±2.63), the number of crossplatform of high-dose group(4.33 ±2.08) was statistically reduced (P＜0.05). B(a)P at 10 and 20 mg/kg decreased NMDAR2B and BDNF expression in hippocampus of rats in immunohistochemistry. The level of NMDAR2B was (162.23 ±6.56) in the high-dose group and (150.38 ± 15.34) in the control group(P＜0.05);the expression level of BDNF was (163. 13 ± 8.09) in the high-dose group and (141.83 ± 13.37) in the control group(P＜ 0.05). Conclusion Subacute B(a)P exposure can reduce spatial learning and memory in weaning rats, it may be related to decreased levels of NMDAR2B and BDNF in hippocampus.

AIM: To investigate mutations of oncogene k-ras in colorectal cancer tissues and the relationship between mutations of k - ras and biological behavior of colorectal carcinoma. METHODS:The specimens of 123 patients with colorectal cancer were collected. Real - time fluorescence quantitative PCR were performed to detect k-ras mutations at codon 12 and codon 13 of exon 1, and the results were analyzed with the corresponding clinical pathological data. RESULTS: Among 123 colorectal cancer cases, point mutations were detected in 53 cases (40.8% ) , point mutations at codon 12 were found in 42 (34.1 % ) cases, and 11(8.9% ) cases at codon 13.No closely relationship between mutations of k-ras and tumor size, location, invasive depth and differentiation extent was observed. The rate of k-ras mutation in the cases with more invaded lymph nodes was higher than that in the cases without invaded lymph nodes ( P < 0.05 ) , and the rate of k-ras gene mutation in the cases with hepatic metastases was higher than that in no hepatic metastases (P <0.05). The rate of k - ras gene mutation was higher in TNM staging Ⅲ/Ⅳ than that in Ⅰ/Ⅱ( P < 0.05 ). CONCLUSION: Mutation of oncogene k-ras plays an important role in the carcinogenesis and development of colorectal cancer, and it is closely associated with invaded lymph notes and hepatic metastases, suggesting that mutation of k- ras indicates a poor prognosis.

Objective: To establish the method of percutaneous endoscopic gastrostomy(PEC) and percutaneous endoscopic jejunostomy (PEJ) for enteral nutrition. Methodes: PEG tubes were placed in 114 patients with Pull method. On the foundation of PEG, PEJ tubes were placed in 26 patients by pushing endoscopy to send tubes through Treitz ligment with usingthe the clip. Results: All PEG insertion was performed successfully. PEJ tubes were placed successfully with a new method in 26 patients. 15 patients had a little blooding and 8 patients had slight infection. 21 patients had respiratory tract infection and had been cured by using antibiotic. There was no severe complication. Conclusion: PEG is simple、safe、efficient. The new method of PEJ is feasible.

Clinical data of 19 patients with congenital pyruvate kinase deficiency were analyzed. Insufficient pyruvate kinase confirmed the diagnosis. Laboratory parameters of hemolysis were summarized. In cases of neonatal hyperbilirubinemia and unexplained hemolytic anemia, pyruvate kinase activity and next generation sequencing test may help the early diagnosis.

Objective: To investigate the relationship between the eosin-5′-maleimide (EMA) binding test and the clinical severity of hereditary spherocytosis (HS). Methods: A total of 258 un-splenectomize HS patients were consecutively enrolled. Correlation of hemoglobin concentration, hemolytic parameters, compensating erythropoiesis and the EMA binding test were evaluated. Results: 258 (128 male and 130 female) patients were included in this study, including 91 compensatory hemolysis patients, 53 patients with mild anemia, 78 patients with moderate anemia and 36 patients with severe anemia. The median age at diagnosis was 23 (2-70) years. The median decreased fluorescence intensity of EMA binding test was 29.97% (16.09%-47.34%) and the average intensity was (29.70±6.28) % of 258 HS patients. The decreased EMA binding fluorescence intensity correlated with MCV (r=-0.343, P<0.001) and MCHC (r=0.223, P<0.001). There was no relationship between EMA fluorescence intensity and absolute reticulocyte count (r=0.080, P=0.198) , reticulocyte percentile (r=-0.015, P=0.813) , IBIL levels (r=-0.009, P=0.902) , HGB levels (r=-0.067, P=0.280). Evaluated as a quartile variable, EMA fluorescence intensity was not correlated with anemia severity (C=0.150, P=0.746). Conclusion EMA binding test does not related to anemia levels and has no major clinical implications for disease severity in HS.

Objective: To evaluate the impact of the targeted next-generation sequencing (NGS) assay for difficult congenital anemias. Methods: Blood Disease Hospital Anemia Panel 2014 (BDHAP-2014) including 217 known genes of congenital anemias was developed. NGS and parental verification were performed for patients who were suspected diagnosed with congenital anaemia from August 2014 to July 2017. Results: A total of 46 patients were enrolled in this study, the clinical suspection were 11 cases Fanconi anemia (FA), 8 cases congenital dyserythropoietic anemia (CDA), 6 cases congenital sideroblast anemia (CSA), 12 cases congenital hemolytic anemia (CHA), 1 case dyskeratosis congenital (DC), 4 cases iron-refractory iron deficiency anemia and 4 cases unexplained cytopenia (Uc), respectively. 28 (60.9%) of 46 patients became confirmed cases after targeted NGS, corresponding to 44 mutations of which 33 were new. 26(56.5%) patients with results of the assay matching to clinical suspection, including FA (5/11, 45.5%), CSA (6/6, 100.0%), CDA (3/8, 37.5%) and CHA (12/12, 100.0%). 2 (4.3%) cases not matching to clinical suspection, including dyskeratosis congenital (DC) was made in 1(2.2%) patients with suspected FA and familial hemophagocytic lymphohistiocytosis (FHL) was made in 1(2.2%) patients with suspected unexplained cytopenia (Uc). In 12 CHA patients, the hemolytic type was further clarified by the NGS. The remaining 18 cases were not clearly diagnosed. Conclusion Targeted NGS assay is of major impact on congenital anemias. The assay should be used routinely in congenital anemias.

Objective: To reveal the genetic characteristics of erythrocyte membrane protein in hereditary spherocytosis (HS) in China. Methods: Next-generation sequencing technology was used to detect mutations in genes of erythrocyte membrane proteins in 51 clinically diagnosed HS patients. The relationship between gene mutations and clinical phenotypes was analyzed. Results: Mutations in erythrocyte membrane protein genes were detected in 37 patients, including 17 with ANK1 mutations (17/37, 45.9%), 14 with SPTB mutations (14/37, 37.8%), and 5 with SLC4A1 mutations (5/37, 13.5%). One patient carried both heterozygous ANK1 mutation and SPTB mutation (1/37, 2.7%). SPTA1 and EPB42 mutation was not fou nd in any patient. Nonsense mutations (36.8%) and missense mutations (31.6%) were most common. Of the 38 mutations detected, 34 were novel mutations and have not been reported elsewhere (89.5%). Sixteen HS patients underwent parental genetic validation, 6 patients (37.5%) inherited gene mutation from parents and 10 (62.5%) were de novo. The peripheral blood cell parameters of HS patients were not related to the mutant genes and gene mutation types. However, it seems that HS patients with mild clinical status are prone to carry SPTB mutations while more patients with severe clinical status have ANK1 mutations. Conclusions ANK1 and SPTB are the most common mutant genes in Chinese HS patients, mainly with missense mutations and nonsense mutations. There was no significant correlation between the mutation of HS related genes and the severity of HS.