Objective To explore the etiology,clinical characteristics,diagnosis and treatment of the bile duct perforation in children.Methods The clinical data of 7 children with the bile duct perforation were retrospectively summarized in Wuhan Children's Hospital from April of 2009 to April of 2014.Results There were 7 cases of the children with perforation of the bile duct,1 male and 6 female,the average age was 2.05 years.The most common presenting symptoms were abdominal distension in 7 cases(100.0％),nausea and vomiting in 6 cases(85.7％),abdominal pain in 5 cases(71.4％),jaundice in 1 case(14.3％) and diarrhea in 1 case(14.3％).Six cases experienced preoperative abdominal paracentesis,which all gained bilious ascites.Both abdominal ultrasound and computed tomography(CT) showed ascites in 5 cases.On exploration,sites of perforation were seen in 3 cases(42.8％) at the junction of the common hepatic duct and cystic duct,1 case(14.3％) at common hepatic duct,and 1 case(14.3％) at common bile duct,while sites of perforation in other 2 cases(28.6％) were not localized.In the cases(case 1,2,5 and 7) whose site of perforation was large,the T-tube drainage and peritoneal drainage through laparotomy or laparoscopic surgery was performed.In case 4 whose site of perforation was very small,and case 3 and 6 whose site of perforation was not localized,the cholysystostomy and peritoneal drainage was performed through laparotomy or laparoscopic surgery.Simple closure of the perforation was performed in case 4.Case 4 and 5 showed recurrent abdominal pain after operation and abdominal CT revealed biliary tract dilatation,and then biliary reconstruction was performed.Both of the patients recovered well postoperatively.The other 5 children recovered well and had an uneventful postoperative period from the 7 months to 5 years follow-up.Conclusions Early diagnosis of perforation of the bile duct can be made based on clinical manifestations,abdominal ultrasound and CT and abdominal paracentesis.Active surgical treatment should be performed once diagnosis was made.Depending on the size of perforation of the bile duct,appropriate surgical drainage scheme is made.The children with recurrent abdominal pain and biliary tract dilatation should receive biliary reconstruction.

The paper summarizes application of new media in patient informatization services in hospitals, introduces exploration and practices made by Children′s Hospital of Shanghai on new media informatization platform construction from such aspects of OA system-basedwindowservice platform,order typeservice model provided to the public onMicroblogandWeChatplatforms andex-press typemobile network terminal service for hospital staffs.The service effects are summarized at last.

ObjectiveTo evaluate the neurobiological characteristics of human histio-amniotic mesenchymal (hAMCs) and effect of hAMCs transplantation into the brain to treat Parkinson's disease(PD) modle mice.MethodsThe expressions of mesenchymal stem cells, neural stem cells, dopaminergic neurons and markers related to neurogenesis such as Vimentin, STRO-1, nestin, CD133, β-tubulin, TH, DAT, Ngn2 and mash-1 in hAMCs were evaluated through immunocytochemical stain; and the mRNA transcriptions of neural stem cell markers, Vimentin and nestin in hAMCs were detected by RT-PCR. The PD model was induced by MPTP(i.p.) in C57BL/6 mice transplanted with hAMCs into the right striatum. The therapeutical effect of hAMCs on PD mice was evaluated by spontaneous movement, rotating bar test and the immunohistochemistry of anti-human chondrosome and TH antibodies in striatum.ResultshAMCs induced by nerve cells culture medium, expressed mesenchymal stem cells, neural stem cells, dopaminergic neurons and other specific markers related to neurogenesis mentioned above. The frequency of spontaneous movement in PD mice was significantly increased(P<0-05), and the time of rotating bar was obviously prolonged(P<0-05) after transplantation with hAMCs.ConclusionhAMCs possess the characteristics of nerve cells after cultured in vitro and can significantly recover the damage of motor function induced by MPTP after transplantation into striatum in PD model mice.

OBJECTIVETo study the chemical constituents of Eriophyton wallichii.

METHODThe constituents were isolated and purified by silica gel column and Sephadex LH-20 chromatography. Their structures are elucidated by physicochemical properties and spectral analysis.

RESULTSeven compounds were obtained and identified as beta-sitosterol (1), marrubiin (2), ursolic acid (3), cimigoside (4), 5-deoxyantirrhinoside (5), 8-epiloganic acid (6) and apigenin 7-(6"-p-coumaroyl) glucoside (7).

CONCLUSIONCompounds 1-7 were isolated from this plant for the first time.

Apigenin ; chemistry ; isolation & purification ; Chromatography, Gel ; Diterpenes ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Iridoids ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

Objective To explore the optimal experiment conditions of CCK-8 and MTS for cell proliferation assays in human amniotic epithelial cells and to evaluate the cytotoxicity of these reagents. Methods Human amniotic epithelial cells (hAECs) in logarithm growth stages were prepared in different cell concentrations with DMEM/F12 and 10% FBS. The sensitivity and optimal wavelengths was determined based on the optical density (OD) measured at 450 nm and 492 nm. The optimal time was determined under the conditions of the same cell concentration and defined OD values. HAECs were treated with DMSO, CCK-8 and MTS for 1 h, 2 h, 3 h, and 4 h, respectively. 24 h later, cytotoxicity of the CCK-8 and MTS was evaluated by determination of cell proliferation and Trypan Blue staining. Results The optimal detection wavelength was 450 nm for CCK-8, and 492 nm for MTS. The sensitivity of CCK-8 was slightly lower then that of MTS. The optimal time for incubation hAECs with CCK-8 was 4 h within 1~4 h. The inhibitory on cell proliferation and cytotoxicity of CCK-8 were weaker then those of MTS. Conclusion CCK-8 is a convenient reagent with low cytotoxicity for detection of the proliferation of hAECs.

OBJECTIVETo investigate the relationship between CCR5 delta32 gene polymorphism and condyloma acuminata.

METHODSWe used polymerase chain reaction to amplify the CCR5 gene fragments in 60 patients with condyloma acuminata and 50 age- and sampling date-matched controls, and compared the difference of genotypes between these two groups.

RESULTSNo genotype difference was found between these two groups.

CONCLUSIONCondyloma acuminata are not associated with genetic polymorphism of CCR5 delta32 gene.

Condylomata Acuminata ; genetics ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Receptors, CCR5 ; genetics

OBJECTIVETo investigate the methylation of CpG island in the SHP-1 gene promoter and its significance in lymphoma. To evaluate the effects of As2O3 on demethylation of SHP-1 in human lymphoma cell line T2 and on proliferation of T2 cells.

METHODST2 cells were treated with AsO3. Methylation specific PCR was used to detected the status of SHP-1 methylation in newly diagnosed lymphoma tissues and the T2 cells. The mRNA and protein expression of SHP-1 were determined by FQ-PCR and Western blot. The expression of phospha-c-kit was examined by Westren blot. MTT and flow cytometry were used to determine the growth and apoptosis in T2 cells.

RESULTST2 cells contained completely methylated SHP-1. Furthermore, there was constitutive c-kit phosphorylation. The expression of SHP-1 was recoverd when the cells exposed to AsO3, and concomitant with increasing SHP-1, a parallel down-regulation of phosphorylated c-kit occurred, so that by day 3 phosphorylated c-kit was barely detectable. As2O3 inhibited the cell growth, and the effects were dose- and time-dependent. As2O3 also increased apoptosis rate of T2 cells in a dose- and time-dependent manner, too, and on the 1, 2, 3 d treatment with AsO3 (2.5 micromol/L), the apoptosis rates were 6.12%, 26.53%, 50.90%, respectively. The frequency of methylation in SHP-1 gene promoter in lymphoma tissues was 87.5% (28/32). In the control group, however, 12 specimens of benign lymph node proliferation showed no methylation in CpG island of SHP-1 gene promoter.

CONCLUSIONHypermethylation of SHP-1 gene promoter in lymphoma indicates the inactivation of SHP-1 gene and its possible role in the tumorigenesis of lymphoma. As2O3 can effectively cause demethylation and inhibit the growth of tumor by reactivating the SHP-1 gene transcription. SHP-1 methylation leading to epigenetic activation of c-kit may have a tentative role in the pathogenesis of lymphoma. Therefore, As2O3 is potentially useful in the treatment of lymphoma as a demethylating agent.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; CpG Islands ; DNA Methylation ; drug effects ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; metabolism ; pathology ; Oxides ; administration & dosage ; pharmacology ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; RNA, Messenger ; metabolism ; Transcriptional Activation ; drug effects ; Up-Regulation

Objective: To explore the expression and prognostic significance of miR-223 in patients with mantle cell lymphoma (MCL) and to investigate the possible mechanism. Methods: Twenty-one newly diagnosed MCL patients with bone marrow involvement were enrolled in the present study, 20 healthy donors as normal control. The expression level of miR-223 and SOX11 mRNA was determined by RQ-PCR. CCK-8 and flow cytometer assays were used to analyze cell proliferation, cell cycle and apoptosis of the constructed miR-223 overexpressing MCL cell line, Granta519 cells. SOX11 protein expression level was determined by Western blot. The target gene of miR-223 was confirmed by dual luciferase reporter assay. Results: ①Of the 21 newly diagnosed MCL patients, 15 were male and 6 female, the median age was 58 (37-72) years. The expression level of miR-223 was significantly down regulated in MCL patients compared with that of healthy donors (14.7±10.5 vs 1 244.1±1 935.2, P<0.001). The lower expression of miR-223 was inversely correlated with high-risk mantle international prognostic index (P=0.001), elevated LDH (P=0.001), ECOG score ≥2 (P=0.035). ②Using the median relative expression level of miR-223 as the cutoff value, 21 MCL patients were divided into high-expression group (n=10) and low-expression group (n=11) and found that the high-expression group had a significantly superior OS (median OS: 36 vs 12 months, P=0.021). ③In vitro results showed that compared with the control group, the proliferation of miR-223 overexpressed Granta519 cells was inhibited (the most significant reduction on 96h, P<0.001), manifested by lower proportion of cells in G2/M phase (P<0.001) and increased apoptosis (P<0.001), and the expression level of SOX11 protein in Granta519 cells was significantly lower than that of the control group. ④miR-223 could inhibited the 3' untranslated region of SOX11, and the expression level of miR-223 was significantly negatively correlated with mRNA level of SOX11 in MCL patients (r=-0.81, P<0.001). Conclusions: The expression of miR-223 was repressed in MCL and was associated with poor clinical outcomes, which may be probably attributed to its direct targeting SOX11.
Adult ; Aged ; Female ; Humans ; Lymphoma, Mantle-Cell ; Male ; MicroRNAs ; Middle Aged ; Prognosis ; RNA, Messenger ; SOXC Transcription Factors

OBJECTIVETo construct the eukaryotic expression vectors of human cyclin D1 gene and express them in poorly differentiated nasopharyngeal carcinoma cells (CNE-2Z cells).

METHODSThe full-length cyclin D1 was cloned from CNE-2Z cells by RT-PCR. The cDNA fragments were inserted into pIRES2-EGFP plasmids and pEGFP-C2 plasmids and confirmed by restriction enzyme digestion, PCR and sequencing. The recombinant vectors were transfected into CNE-2Z cells via Lipofectamine 2000, and the expression of cyclin D1 in the cells was examined by immunofluorescence and Western blotting.

RESULTSAgarose gel electrophoresis showed a 918 bp band of the RT-PCR products, which matched the expected size. Restriction enzyme digestion, PCR and sequencing demonstrated successful construction of the recombinant vectors. CNE-2Z cells transfected with the recombinant vectors expressed cyclin D1 protein or cyclin D1-GFP protein as were verified by immunofluorescence and Western blotting.

CONCLUSIONWe have cloned cyclin D1 gene and constructed its eukaryotic expression vectors that can be expressed in nasopharyngeal carcinoma cells, which may facilitate the study of the role of cyclin D1 in the development of nasopharyngeal carcinoma.

Cell Line, Tumor ; Cloning, Molecular ; Cyclin D1 ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVEThe purpose of this study was to investigate the clinicopathologic factors associated with false-negative rate of sentinel lymph node biopsy (SLNB) in breast cancer, and to explore how to reduce the false-negative rate of SLNB.

METHODSThe clinicopathological data of 2265 patients with invasive breast carcinoma who underwent sentinel lymph nodes biopsy (SLNB) in Shandong Cancer Hospital between November 1999 and December 2011 were retrospectively analyzed. We screened 1228 patients who received axillary lymph node dissection after SLNB, and studied the clinicopathological factors that could be associated with false-negative rate of SLNB.

RESULTSThe false negative rate of this group was 10.7% (73/683), accuracy rate was 94.1% (1155/1228), and negative predictive value was 88.2% (545/618). Clinical tumor size (all P < 0.05), calendar year of surgery (all P < 0.05) and numbers of detected SLNs (all P < 0.05) were significantly related with false negative rate and accuracy rate of SLNB, determined by single factor analysis. Logistic regression model analysis showed that calendar year of surgery (P = 0.034) and numbers of detected SLNs (P = 0.012) were independent predictive factors for the false negative rate of SLNB.

CONCLUSIONSFalse negative rate and accuracy rate of SLNB are significantly related to the calendar year of surgery and number of detected SLNs. Strict case selection, standard operation procedure, increaseing numbers of detected SLNs, and improvement of the skill of operators are effective measures to reduce the false negative rate of SLNB.

Adult ; Aged ; Axilla ; Breast Neoplasms ; pathology ; surgery ; Carcinoma, Ductal, Breast ; pathology ; surgery ; Carcinoma, Lobular ; pathology ; surgery ; Carcinoma, Medullary ; pathology ; surgery ; False Negative Reactions ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Middle Aged ; Retrospective Studies ; Sentinel Lymph Node Biopsy ; Young Adult