Objective　To introduce the development of therapy of hilar cholangiocarcinoma. Methods　This summarization paper was made on the literatures review. Results　Extended radical resection, neoadjuvant chemotherapy, orthotopic liver transplantation, photodynamic therapy and molecular chemoradiotherapy might improve the survival rate. Conclusions　Surgical resection combined with other theraputic methods is the main treatment for hilar cholangiocarcinoma.

Objective To observe the effect of IL 11 on graft versus host disease and graft versus leukemia after allogenic bone marrow transplantation and related mechanism in acute lymphoblastic leukemic (ALL) mice. Methods Twenty nude mice and 20 BABL/c mice were randomly divided into 2 groups separately, total 4 groups. Group A and C, nude mice and BABL/c mice control group respectively; Group B and D, nude mice and BABL/c mice experimental group. The mice in the experimental groups were subjected to the subcutaneous injection of IL 11 2 days before transplantation and 7 days after transplantation, while those in the control groups not to IL 11. The effects of IL 11 on CD4 + and CD8 + T cells before and after transplantation in ALL mice were evaluated by flow cytometry. The survival time of ALL mice after marrow transplantation was recorded. The GVHD pathological changes of liver, spleen and intestine in ALL mice after transplantation were observed. The level of tumor necrosis factor ? (TNF ?) was detected by ELISA. Results In group D, IL 11 could significantly decrease the number of CD4 + T cells and increase CD8 + T cells simultaneously. IL 11 could obviously prolong the survival time, delay and relieve GVHD, and decrease the level of TNF ? in ALL mice after transplantation. Conclusion IL 11 could alleviate GVHD and retain GVL effect after allogenic bone marrow transplantation.

Copy number variations in the human genome,one of the causes of complex diseases and genetic diseases,can lead to genomic disorders.As these diseases are difficult to diagnose,it is significantly meaningful to conduct genetic researches and molecular diagnosis.Chromosomal microarray can be used to detect copy number variations on a genome-wide scale.With the advantage of high throughput and resolution,chromosomal microarray is perceived as an important means of identifying copy number variations in genomic disorders.As technology advancements of chromosomal microarray and accumulations of clinical experiences,chromosomal microarray has played a significant role in etiological diagnosis of multiple malformations,mental retardation and autism.

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) singling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by zinc sulfate preconditioning in rats.Methods SPF healthy male Sprague-Dawley rats,weighing 250-280 g,aged 16-20 weeks,were used in this study.After the animals were anesthetized,their hearts were immediately removed and retrogradely perfused with an oxygenated K-H solution at 37 ℃ in a Langendorff apparatus.Forty isolated rat hearts were randomly divided into 4 groups (n=10 each):control group (group C),I/R group,zinc sulfate preconditioning group (group Zn) and zinc sulfate preconditioning plus Nrf2/ARE singling pathway blocker luteolin group (group Zn+Lut).After 20 min of equilibration,the hearts were continuously perfused for 100 min in group C,the hearts were perfused with 4 ℃ St.Thomas′ cardioplegic solution before ischemia and then subjected to 40 min global ischemia followed by 60 min reperfusion to establish the model of I/R in group I/R,200 μmol/L zinc sulfate 1.5 ml/kg was injected intraperitoneally,and 24 h later the model of I/R was established in group Zn,and in group Zn+Lut,the hearts were perfused for 3 min with K-H solution containing luteolin 50 μmol/L starting from the time point immediately before ischemia,and the other treatments were similar to those previously described in group Zn.Heart rate (HR),left ventricular end diabetic pressure (LVEDP),left ventricular developed pressure (LVDP) and the maximum rate of increase of left ventricular pressure (+dp/dtmax) were recorded at the end of equilibration and reperfusion.Coronary effluent was collected at the end of reperfusion to measure the levels of lactate dehydrogenase (LDH) and malondialdehyde (MDA).The expression of heme oxygenase-1 (HO-1),quinone oxidoreductase (NQO1),superoxide dismutase 1 (SOD1) and Nrf2 in myocardial tissues was detected by Western blot.Results Compared with group C,HR,+dp/dtmax and LVDP were significantly decreased,LVEDP was increased,and the levels of LDH and MDA in coronary effluent were increased at the end of reperfusion in I/R and Zn+Lut groups,and the expression of NQO1,HO-1,Nrf2 and SOD1 was up-regulated in I/R,Zn and Zn+Lut groups (P<0.05).Compared with group I/R,the HR,+dp/dtmax and LVDP were significantly increased,LVEDP was decreased,and the levels of LDH and MDA in coronary effluent were decreased at the end of reperfusion,and the expression of NQO1,HO-1,Nrf2 and SOD1 was up-regulated in group Zn (P<0.05).Compared with group Zn,HR,+dp/dtmax and LVDP were significantly decreased,LVEDP was increased,and the levels of LDH and MDA in coronary effluent were increased at the end of reperfusion,and the expression of NQO1,HO-1 and SOD1 was down-regulated(P<0.05),and no significant change was found in Nrf2 expression in group Zn+Lut (P>0.05).Conclusion The mechanism by which zinc sulfate preconditioning reduces myocardial I/R injury is related to activation of Nrf2/ARE singling pathway in rats.

Objective To study the treatment of intra-abdominal hypertension syndrome (IAHS) secondary to fulminant acute pancreatitis(FAP). MethodsWe retrospectively analyse therapeutic results of 14 IAHS cases secondary to FAP during the period of 1998～2003. ResultsFour out of the 6 cases receiving nonoperative therapy died with motality rate of 67.7% (4/6). Two out of the 8 cases treated by early surgery died. The motality rate was 25% (2/8), which was significantly different from that treated conservatively . ConclusionModerate to severe IAHS cases secondary to fulminant acute pancreatitis should undergo exploration in the early phase of disease to improve the prognosis of FAP.

Objective To evaluate the value of PCR-restriction fragment length polymorphism (PCR-RFLP),real-time PCR and multiplex ligation-dependent probe amplification (MLPA) in the genetic diagnosis of spinal muscular atrophy (SMA) and make laboratory support accessible to clinicians for the molecular diagnosis of SMA.Methods Methodological evaluation.Forty-one suspected SMA cases and 359 control individuals received in Shanghai Children's Medical Centre from March 2013 to June 2014 were detected for the deletion of exon 7 and 8 in the survival motor neuron gene 1 (SMN 1) by PCR-RFLP,realtime PCR and MLPA,respectively.Then the results of the three methods were compared and the benefits and limitations of the three methods were evaluated.Results The result of real-time PCR was in complete agreement with that of MLPA,showing that 29 suspected cases harbored homozygous deletions of SMN1 and 1 case possessed heterozygous deletion.Among the homozygous deletions,27 patients demonstrated absence of exon 7 and 8,and 2 cases demonstrated only the absence of exon 7.Meanwhile,both PCR-RFLP and MLPA analysis showed the same results that only 5 out of 395 control cases carried heterozygous deletion.As for cases without heterozygous deletions,PCR-RFLP demonstrated the same result with real-time PCR and MLPA but it missed all the heterozygous ones.Conclusions PCR-RFLP,the conventional SMA gene diagnosis method,was only capable of detecting homozygous deletion of exon 7 and/or 8 of SMN1,but was not as sensitive as to find out the carriers with heterozygous deletions.MLPA could detect the deletion and quantify the copy numbers of exon 7 and 8 of SMN1,efficiently,while the price was relatively high,which brings challenges for its application in the carrier screening of SMA.Compared with these two methods,realtime PCR with high throughput and low input was a rapid,acourate and economic method for the genetic diagnosis of SMA and carrier screening in large populations.

Objective To investigate the possibility and feasibility of the whole genome microarray scanning technique in clinical cytogenetic diagnosis of an uncertain karyotype and mentally retarded child. Methods The karyotype analysis of the mental development delayed child was 47, XY+mar. Genomic DNA was extracted from the peripheral blood and the whole genome microarray scanning technique was used to analyze the derivative chromosome. Results The whole genome microar-ray scanning technique indicated the derivative chromosome fragment had originated from 9p13.1-p24.3. Conclusions Com-paring to conventional cytogenetic analysis methods, the whole genome microarray scanning technique is of high resolution, high-throughput and high accuracy, which can detect the submicroscopic chromosomal aberrations and replace the conven-tional karyotype analysis.

Objective To establish a method of gene mutation detection for congenital adrenal hyperplasia (CAH) by using sequencing, single nucleotide polymorphisms (SNP) analysis and T-A cloning. Methods The blood samples of 33 patients with 21-hydroxylase deficiency (21-OHD) , 2 patients with 17α-hydroxylase deficiency (17-OHD) , the parents of all the patients and 105 healthy children were collected. Genomic DNA were extracted form the blood samples. To detect the gene mutation of CYP21A2,highly specific primers for CYP21A2 gene were designed according to the sequence differences between CYP21A2 gene and its pseudogene. The whole CYP21A2 gene was amplified and sequenced. SNP analysis and TA cloning of PCR products were also carried out. The molecular diagnosis of 17-OHD was based on the amplification and sequencing of CYP17A1 gene. Results The corresponding gene mutations was determined in all the patients based on the method established in this study. Thirteen mutations of CYP21A2 gene were identified in 33 patients with 21-OHD. The 3 most frequent mutation of CYP21A2 gene were IVS2-13A/C >G, p. I172N and chimeric mutation, which accounted for 32% (21/66) ,27% (18/66) and 15% (10/66) respectively. Ninety-one persent mutations of CYP21A2 gene resulted from pseudogene conversion. In 2 patients with 17-OHD, homozygous mutations of CYP17A1 gene, IVS4-6A > G and p. 487_489del were identified separately. All the gene mutations detected in the patients were inherited from their parents. No mutation of CYP21A2 gene or CYP17A1 gene was found in the healthy children. Conclusion A method of gene mutation detection for CAH has been established. It will be beneficial to clinical diagnosis of CAH.

OBJECTIVE To analyze the distribution and anti-microbial resistance profile of pathogenic bacteria isolated from hospitalized neonates in Wuhan area.METHODS The strains of bacteria isolated from neonates in hospitalization of Wuhan area from 2006 to 2007 were identified by VITEK 32 automatic bacteria system and antimicrobial susceptibility test was performed by Kirby-Bauer method.The standards of NCCLS issued in 2006 were used to assess the results of antimicrobial susceptibility.RESULTS A total of 3892 strains of bacteria were isolated from 10053 specimens,the positive rate was 38.7%.The Gram-positive cocci and Gram-negative bacilli accounted for 41.0% and 55.2%,respectively.The leading 8 species on the list of isolated strains in the two years accounted for 90.8% of all the isolated strains.MRCNS accounted for 68.6% of coagulase negative staphylococcus,and the MRSA kept a low rate(2.6%) in Staphylococcus aureus.The infection rate of newborn by Enterococcus in blood was remarkably higher than that of the past report in this area.The ?-lactamases-producing Escherichia coli and Klebsiella pneumoniae were accounted for 46.9% and 34.7%,respectively.CONCLUSIONS The distribution and anti-microbial resistance of hospitalized neonates in Wuhan area have regional and group characteristics;it′s necessary to strengthen monitoring the regional epidemic characteristic status and the drug-resistant status of the pathogenic bacteria among neonates.

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/ antioxidant response element (ARE) signaling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by ischemic preconditioning in the rats.Methods Healthy male Sprague-Dawley rats,weighing 250-300 g,aged 4-6 months,were used in the study.Their hearts were excised,and retrogradely perfused with K-H solution at 37 ℃ in a Langendorff apparatus.Forty isolated hearts were randomly divided into 4 groups (n=10 each) using a random number table:control group (group C),I/R group,ischemic preconditioning group (group IPC),and ischemic preconditioning +Nrf2/ARE signaling pathway blocker luteolin group (group IPC+L).After 20 min of equilibration,the hearts were continuously perfused for 100 min in group C.After 20 min of equilibration,the hearts were subjected to 40 min ischemia at 32 ℃ followed by 60 min of reperfusion in group I/R.In group IPC,ischemic preconditioning was induced by 6 cycles of 10 s ischemia followed by 10 s reperfusion starting from the time point immediately after 20 min of equilibration,and then the hearts were subjected to 40 min ischemia at 32 ℃ followed by 58 min of reperfusion.In group IPC+L,after 20 min of equilibration,the hearts were perfused with K-H solution containing lueolin 50 μmol/L for 3 min before ischemia,and the other treatments were similar to those previously described in group IPC.Left ventricular developed pressure (LVDP),left ventricular end-diastolic pressure (LVDEP),heart rate (HR),and the maximum rate of increase of left ventricular pressure (+dp/dtmax) were recorded at the end of equilibration and reperfusion.At the end of reperfusion,left ventricular myocardial tissues were obtained for examination of the ultrastructure of myocardial cells and for determination of the expression of Nrf2,heme oxygenase-1 (HO-1),quinone oxidoreductase 1 (NQO1),and superoxide dismutase 1 (SOD1) mRNA and protein (by real-time polymerase chain reaction and Western blot,respectively).Results Compared with group C,the HR,+ dp/dtmax and LVDP were significantly decreased,and LVEDP was significantly increased at the end of reperfusion in I/R and IPC+L groups,and the expression of Nrf2,HO-1,NQO1 and SOD1 mRNA and protein was significantly up-regulated in I/R,IPC and IPC+L groups (P＜0.05).Compared with group I/R,the HR,+dp/dtmax and LVDP were significantly increased,and LVEDP was significantly decreased at the end of reperfusion,the expression of Nrf2,HO-1,NQO1 and SOD1 mRNA and protein was significantly up-regulated (P＜0.05),and the pathological changes were significantly attenuated in group IPC,and no significant change was found in the parameters mentioned above in group IPC+L (P＞0.05).Compared with group IPC,the HR,+dp/dt and LVDP were significantly decreased,and LVEDP was significantly increased at the end of reperfusion,and the expression of HO-1,NQO1,SOD1 mRNA and protein was significantly down-regulated (P＜ 0.05),no significant change was found in Nrf2 mRNA and protein expression (P＞0.05),and the pathological changes were significantly aggravated in group IPC + L.Conclusion Ischemic preconditioning reduces myocardial I/R injury through activating Nrf2/ARE signaling pathway in the rats.