Objective To evaluate the significance and expression of the GDNF/GFRα1/RET genes in distal rectum with Congenital Anorectal Malformations(ARMs) Methods Specimens were collected from resected colon which is 3 cm away to the anus and the distal of rectum in 12 ARMs patients. Haematoxylin and Eosin (HE) staining, immunohistochemical (IHC) staining and reverse transcription PCR (RT-PCR) were used to test RET, GDNF and GFRα1 expression in the differ-ent site of samples with ARMs. Results Expression of ganglia and RET, GDNF and GFRα1 were all negative in distal rec-tum in 12 ARMs patients. Ganglionic cells were found in tissues 3 cm away from the anus where RET, GDNF and GFRα1 expression were also positive in those ARMs patients. Expression of RET, GDNF and GFRα1 were strong positive in middle and low imperforate anus indicated by brown color and week positive in high imperforate anus indicated by yellow color. One case of faeces contamination and one case of loose motion without anal incontinence were found in post-op follow up of high ARMs patients. By contrast, one case of rectal mucosa prolapse and one case of occasional faeces contamination which recov-ered with hip bath were found in post op follow up in middle or low ARMs. Conclusion The unsatisfactory anorectal func-tion is possibly related to the decrease or lost of neurotrophic factors (GDNF/GFRα1/RET) and ganglia in the myenteric plex-uses post plastic surgery in ARMs patients.

Objective To investigate the relationship between the methylation of RET (proto-oncogene, RET) and Hirschsprung disease (HD), and understand its significance in the development of intestinal wall ganglion cells. Methods Twenty-one surgical removal specimens, which were all dilation segment of HD in Tianjin Children’s Hospital, were used as experimental group, and 5 samples of non-HD normal colon tissues were used as control group. The bisulfite sequencing (BSP)-direct detecting method was used to detect RET CpG island methylation status. The expression of RET protein was detected by immunohistochemistry in experimental group and control group. Results In the experimental group 12 cases (57.14%) were found methylation, but no methylation was found in control group. The average optical density of methylated RET protein was 0.201±0.015 in 12 cases. The average optical density of un-methylated RET protein was 0.364±0.023 in 9 cases (P<0.05). Conclusion RET CpG island methylation reduced protein expression levels of RET. The corollary RET gene methylation may influence the expression levels of RET protein, thereby affecting the ganglion cell development, and thus participating in the occurrence of HD.

Objective To explore the role of trypsinogen activation peptide( TAP)in the early di-agnosis of severe acute pancreatitis( AP)and the prediction of outcome. Methods Eighty-nine cases with pancreatitis were collected containing 45 cases of severe AP and 44 of mild AP. 32 cases of non-AP acute abdominal disease were chosen as control group. Blood of all patients was collected within 6h to measure TAP. ROC curve was used to analyze the results. Results The TAP was different among the three groups (p<0. 01);the TAP of patients with severe AP was the highest. ROC curve indicated that 2. 78 nmol/L was the best cutoff to define AP and non-AP;the sensitivity,specificity and accuracy was 88. 8%,100%and 91. 5%,respectively. 8. 55 nmol/L was used to define severe AP and mild AP;the sensitivity,speci-ficity and accuracy was 92. 9%,95. 2% and 88. 7%,respectively. 11. 20 nmol/L was used to predict the occurrence of complications;the sensitivity,specificity and accuracy was 75. 0%,90. 4% and 82. 2%,re-spectively. Conclusion The TAP can be used to diagnose AP,especially to identify severe AP. The TAP is also beneficial for predicting the outcome of patients with severe AP.

The early screening and early diagnosis of biliary atresia is the key to improve the 5-year native liver survival rate,jaundice-free-native-liver survival rate and overall survival rate.So far,the neonatal jaundice,especially that caused by surgical cholestasis exists the lack of awareness.Because of this reason,the delay depletion jaundice patients did not receive early diagnosis and treatment until the severe complications happened.So,it has often missed the best period of operation for the biliary atresia.Called for local medical health governments to formulate the effective early screening methods of biliary atresia and the reasonable schedule of postnatal follow-up,and improving the ability of basic unit medical staff in well understanding cholestasis disease in order to achieve early diagnosis and treatment.Consequently,it will improve the situation of diagnosis and treatment of biliary atresia in our country.

Myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs) are important inhibitory immune cells.Research finds that MDSCs and Tregs increase in a large number under the conditions of inflammation,infection and tumor and they are able to suppress the immunity system and promote tumorigenesis and metastasis through multiple mechanisms.Finding the causes resulting in the increase of MDSCs and Tregs in tumor microenvironment and the eliminating methods have become new focuses in the field of antitumor immunotherapy.

The Balanced Score card ( BSC) is a new set of performance measurement and strategic management tools. Based on the basic theory of BSC, and on the basis of establishing the basic framework for performance evaluation of four dimensions, namely satisfaction, financial, internal operation, and growth and development, this paper systematically designs specific evaluation indexes from the 4 dimensions in order to construct the performance evaluation index system for catastrophic disease insurance. The objective of this study is to provide a more scientific and reasonable reference for the operational performance evaluation of catastrophic disease insurance offered to urban and rural residents.

Objective To evaluate the inhibitory activity of Herba Taraxaci extract on Escherichia coli DH5α (E. coli DH5α) and to investigate proteomic response of E. coli. Methods Medicinal powder of Herba Taraxaci was extracted with the solvents of different polarity ( n-hexane, ethyl acetate, distilled water) , and then the obtained 8 different extracts were subjected to thin layer chromatography ( TLC) analysis. Microdilution method was performed to detect the minimum inhibitory concentration ( MIC) of different extracts and the growth curves were described. The protein expression profiles of E . coli treated with the extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis ( SDS-PAGE) and two dimensional electrophoresis (2-DE) . Results Water decoction of Herba Taraxaci could obviously suppress the growth of E. coli with a MIC of 1.95 mg/mL. The different extractions exhibited no antibacterial activity except ethyl acetate phase 3 with a MIC of 0.13 mg/mL, which was equal to 19.23 mg/mL of crude drugs. The results of TLC analysis showed that chlorogenic acid was undetectable in n-hexane extract and ethyl acetate phase 1 extract, and ethyl acetate phase 2 and 3 extracts showed obviously increased spots. The results of SDS-PAGE and 2-DE showed that water decoction of Herba Taraxaci had inhibitory effect on the expression of functional protein. The results of 2-DE showed that after treatment with ethyl acetate phase 3 at the concentration of 2 × MIC for 21 hours, the amount of protein spots were 92 less than those of the blank control group, the spots of E. coli DH5α soluble protein with expression amount down-regulated doubly were 24, and those with expression amount up-regulated doubly were 19. Ethyl acetate phase 3 extract had an effect on down-regulating the protein expression of E. coli DH5α soluble protein pH3-10, and water decoction of Herba Taraxaci had inhibitory effect on E. coli DH5αprotein expression. Conclusion Herba Taraxaci has significant antibacterial activity on E. coli DH5α, and the water-soluble fraction of chlorogenic acid and caffeic acid might be the active components. The possible antibacterial mechanism may be related with the regulation of bacterial protein expression.

Objective To explore the relation between clinical-pathological features,Siewert classification and prognosis of esophagogastric junction (EGJ) carcinoma,and to assess the applicability of the new edition of American Joint Committee of Cancer (AJCC) staging guideline on EGJ adenocarcinoma in China.Methods From 2002 to 2012,the clinical data,pathological features,treatment and prognosis of 218 patients with EGJ malignant tumor were retrospectively analyzed.The patients were typed according to Siewert classification criteria and each case was staged according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma and gastric cancer.Kaplan-Meier method and Log-rank test were performed for survival analysis.Results According to the Siewert classification,type Ⅰ was rare (nine cases,4.1％),type Ⅱ was the most common type (150 cases,68.8％) and followed by type Ⅲ (59 cases,27.1％).There was no significant difference in survival curve among the three types (P＞0.05).The survival curve was drawn according to 7th edition of AJCC TNM staging criteria for esophagus adenocarcinoma.In T staging,the prognosis of patients at T4b was better than that of patients at T4a,the prognosis of patients at ⅡB was better than that of patients at ⅡA.The survival curve of patients at Ⅲ C obviously crossed with that of patients at Ⅳ,which was not in conformity with clinical results.The survival curve was drawn according to 7th edition of AJCC staging criteria for gastric cancer.In T staging,the survival curve of patients at Tis was overlapped with that of patients at T1a.The survival rate of patients at ⅡB could not be accurately predicted by the overall staging.In general,the survival of patients with EGJ carcinoma was better predicted according to 7th edition of AJCC staging criteria for gastric cancer than 7th edition for esophagus adenocarcinoma.Conclusions Neither 7th edition of AJCC staging criteria for esophagus adenocarcinoma nor for gastric cancer could accurately predict its prognosis.In our country,EGJ malignant tumor was similar to gastric cancer and had specific clinical-pathological features.It is necessary to research and establish EGJ carcinoma staging criteria instead of applying the current staging criteria for esophagus adenocarcinoma or gastric cancer.

Objective To explore the histopathological features of 850 patients with esophageal malignant tumor in 10 years.Methods From January 2002 to January 2012, 850 patients diagnosed with esophageal malignant tumor were enrolled.Tumor location, general type, pathological type and TNM stage were retrospectively analyzed.All the data were described as case number and percentage.Results Among the 850 cases of esophageal malignant tumor, 33 lesions (3.9％) located in the neck segment of esophagus, 119 lesions (14.0％) located in the upper segment, 44 lesions (5.2 ％) located in the upper-middle segment, 409 lesions (48.1％) located in the middle segment, 123 lesions (14.5 ％) located in the middle-lower segment, 122 lesions (14.4％) located in the lower segment.Among the 724 eases clearly diagnosed as esophageal malignant tumor by general type, the most cases were ulcer type (305 cases, 42.1％), followed by medulla type (260 cases, 35.9％), fungating type (80 cases, 11.0％) and constrictive type (70 cases, 9.7％), and the least cases were intraluminal type (nine cases, 1.2％).Among the 850 cases of esophageal malignant tumor, squamous cell carcinoma (794 cases, 93.4 ％) was the most common cytological type, followed by small cell carcinoma (19 eases, 2.2％), and the least common cytological type was adenocarcinoma (seven cases, 0.8 ％).Among the 724 cases with clear TNM staging, case number of Tis, T1, T2, T3 and T4 stage was eight (1.1％), six (0.8％), 271 (37.4％), 278 (38.4％) and 161 (22.2％), respectively.Among the 122 cases of distal esophageal carcinomas (104 cases with clear TNM staging), most cases were squamous cell carcinoma (112 cases, 91.8 ％), the others cases were adenocarcinoma (three cases, 2.5 ％), small cell carcinoma (three cases, 2.5 ％), basaloid squamous cell, adenosquamous, neuroendocrine carcinomas and carcinosarcoma (one case in each type, 0.8％).Conclusions Esophageal carcinoma was mostly located in the middle segment of in which squamous cell carcinoma was predominant while adenocarcinoma was less common.Esophageal cancer located at lower segment of esophagus is with a wide range of pathological spectrum, squamous cell carcinoma was still dominant, however, esophageal adenocarcinoma is rare.

This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.