BACKGROUND: Biliary complications are a common cause of death in patients after liver transplantation, biliary endoscopic minimally invasive technique is gradually becoming an important tool to solve biliary complications following orthotopic liver transplantation.OBJECTIVE: To analyze the literatures on the biliary tract complications after liver transplantation, and to demonstrate a clear role of bile duct endoscopy in biliary complications.METHODS: Using "orthotopic liver transplantation, biliary complications, biliary tract, endoscope technique" in English for the search term, Pubmed database were searched between January 1980 and October 2008; Using "liver transplantation, biliary complications, bile duct cast, cholangioscopy" in Chinese for the search term, Vip Chinese Periodical Database and CNKI database were searched between 1994 and January 2009. Literature language was limited to English and Chinese. The studies related to bile duct injury-caused biliary complications after liver transplantation were included, while other research unrelated to biliary complications after liver transplantation were excluded.RESULTS AND CONCLUSION: A total of 52 literatures were screened out by the primary computer inspection, according to inclusion and exclusion criteria, 30 ones were involved for analysis. Biliary complications and vascular complications are considered as a common cause of death in patients after liver transplantation, particularly in biliary complications has become the major reason limiting the development of liver transplantation. Because of the difficulties on early recognition and treatment, the importance of the treatment for the complications in liver transplant patients is increasingly attracting more and more attention. Endoscopic operation technology and their subsidiary parts are improving, particularly therapeutic endoscopic retrograde cholangiopancreatography and biliary mirror technology are developing and continuously improving, biliary endoscopic minimally invasive technique is gradually becoming a major approach to solve biliary complications after orthotopic liver transplantation, The minimally invasive endoscopic diagnosis and treatment is an intuitive, reliable and credible means for the biliary complications after liver transplantation, serving as the most preferred method for diagnosing and treating biliary complications after liver transplantation.

Objective To screen polymorphisms and mutations in the promoter region of WD gene.Methods DNA from peripheral blood was obtained from 71 subjects of 36 family (48 WD patients,23 patients first-degree relatives) and 20 healthy people from Feb.2001 to Feb.2004.DNA sequence of the genes was analyzed by PCR amplification and direct genomic sequencing.Results There were three polymorphisms at positions-190,-78,+260(transcription start site as +1) of the promoter region of WD gene.Normal controls,WD patients and patients’ first-degree relatives all showed the polymorphisms;three of 48 WD patients presented C→T base substitution mutations at the same position -183:two were homozygous mutation,while the other was heterozygous.Normal control subjects and patients' relatives didn’t show this kind of mutation.Conclusion It suggests that the mutation of the promoter region is one of WD pathogenesis.

Objective To study genotyping and molecular epidemiology distribution of GBS pathogenic strains of GBS positive pregnant women in Guangzhou,for GBS pathogenic strains of rapid molecular diagnosis and epidemiological surveillance pro-vide certain theoretical basis and method.Methods In the Guangzhou area,used multi stage stratified sampling method col-lecting GBS positive pregnant women’s reproductive tract specimens from January to December 2015,drug sensitivity quality control standard strains:Streptococcus pneumoniae (ATCC49619)and Staphylococcus aureus (ATCC25923),took culture of bacterial,strain,identification,DNA extraction,PCR,gene detection method,through the relevant software for data analy-sis,analyzed GBS strains of gene and molecular epidemiology.Results In the study,collected 2 812 samples of secretions,af-ter identification of strains isolated from 178 strains of pathogenic GBS strains,the detection rate was 6.33%.GBS patho-genic strains to linezolid vancomycin,penicillin,nitrfurantion and other antimicrobial drug resistance rate was 0,GBS parho-genic strains to ampicillin,ciprfloxacin moxifloxacin and levofloxacintesistant parts,the restance rates were 1.1%,16.9%, 18.0% and 22.5%,but GBS pathogenic strains to erythromycin,clindamycin tetracydine antibiotics showed a high resistance rate,the resistance rates were 50.6%,47.8%(of which 20 cases of erythromycin induced clindamycin resistance accouted for 23.5%)and 73.0%.Among them,65 strains of GBS detected the mreA gene,56 strains of GBS detected the ermB gene,36 strains of GBS detected the mefA gene,28 strains of GBS detected the mefE gene,5 strains of GBS detected the ermA gene, ermC gene was not detected in the gene.Among them,carried five multidrug resistance gene of 3 strains (1.6 9%)and 4 kinds of resistant gene carried with 15 strains (8.43%),carried three resistance genes of 19 strains (10.67%),2 kinds of resistant gene carrying a 25 strains (14.04%),carried the resistance gene of 5 strains (2.81%),did not carry resistance gene of 1 strain (0.56%).The nucleotide sequences of the five drug resistance genes were 100%,and no gene mutation oc-curred.Conclusion The main GBS disease resistant gene was mreA,ermA,ermB,mrfA,mefE and its nucleotide sequence homology was 100%.The clinical need to strengthen the detection of resistant gene and molecular level and guide clinical more scientific and rational drug use.

Objective 　To study the sequence and structure of intron 8 in WD gene in order to further understand the relationship between intron 8 and WD. Methods　We utilized polymerase chain reaction (PCR) to the amplification of exon 8-intron 8-exon 9 which were then sequenced by a dideoxy chain termination methon in 10 normal controls and 32 members of 11 families(20 WD patients and 10 of their relative). The results were analyzed by the computer. Results　The sequence of intron 8 was 703 bp with the G　+　C content of 42.7%. There were one short tandom repeats, 7 direct and inverted repeats in it. An open reading frame coded with 82aa was found at 323 base pairs of downstream of a TATAbox. There were two DNA polymorphisms at 408 and 487 nucleotides. The sequence analysis showed that the 5end has the sequence of 5-GTAAC, 3end has the sequence of CCTAG-3, and branchpoint of 5-TTTCGA-3.Conclusions　The sequences and structures of intron 8 in WD familiess members are not different from normal controls. Our data suggest that the WD gene intron 8 might not play an important role in the pathogenesis of WD.

ObjectiveTo explore the diagnostic and predictive value of troponin Ⅰ,lactic acid and hypersensitive C-reactive protein(hs-CRP) in viral myocarditis.MethodsTroponin Ⅰ,hs-CRP,lactic acid blood levels in different time were measured in 88 patients with acute viral myocarditis(54 cases) or acute upper respiratory infections (34 cases).Troponin Ⅰ leyel in patients with severe 39 cases and mild VMC 15 cases were compared.ResultsTroponin Ⅰ level of VMC group was (0.59 ±0.10) ng/L,and significantly higher than that of acute upper respiratory infection group [ (0.10 ± 0.08 ) ng/L ] ( t =2.79,P ＜ 0.05 ).And higher lactic acid and hs-CRP level were observed at different period ( t =2.71,2.48,all P ＜ 0.05).ConclusionTroponin I could help diagnose VMC predict the severity of myocarditis to some extent.Lactic acid and hs-CRP could also reflect imflammation injury of VMC.

Objective 　Determination of Wilson disease gene mRNA expression in human fibroblast cell strain (Me32aT22/2L) by reverse transcription-polymerase chain reaction (RT-PCR). Methods　Using lipofection reagent, the plasmid vector carrying the Wilson disease gene (pRc/CMV-WD) was transferred into Me32aT22/2L cultured in serum free complement medium. RT-PCR was used to determine WD mRNA expression in Me32aT22/2L. Results　 Wilson disease gene expression was detected in Me32aT22/2L, while no specific signals were detected in untransfected fibroblast. Conclusions　It demonstrated that Me32aT22/2L strain could express the Wilson disease gene, suggesting that Wilson disease gene transfer might develop a new approach to study Wilson disease.

Objective To carry out a taxonomic identification of a strain of claviform bacteria iso-lated from prostatic fluid of a patient who suffered from chronic prostatitis, and to approach its phylogenic and biologic position. Methods We undertaked an initial identification by phenotypic characters such as morphologecal, physiological and biochemical characteristics to ascertain its phylogeny by chemical composi-tion analysis of cell wall and 16S rRNA gene sequencing and alignment. Results A club-shaped gram posi-tive rod bacillus was isolated in pure culture state. Its biochemical reactions were not active. The diamino-acid of cell wall was meso-diaminopimelic acid (meso-DAP) and it had wall chemotype Ⅳ ( contained arabi-nose, galactose and maltose ). Sequence searches of the GenBank database revealed that this strain had a highest level of 16S rDNA sequence similarity (99.4%) to C. tuberculostearicurn strain ATCC35692 with only 8 nucleotides difference. Conclusion On the basis of phenotypic and phylngenetie analysis, it is rea-sonable to assign this strain to the species C. tuberculostearicum, and this is the first isolation of C. tubercu-lostearicum from prostatic fluid home and abroad.

Objective To find out and clone the genes that can influence different willis circle in Meriones unguiculatus, the genomic library has been made.Methods 96 clean Meriones unguiculatus were dissected according to observe the differences of willis circle, and the mixed DNA was extracted from different blood vessels.The genomic library construction has been done by using pCC1FOS vector, following by CopyControl Fosmid Lib manual.The copy numbers, recombined segment size and recombination fraction of the library were measured.Results Genomic library of different Willis circle in Meriones unguiculatus was successfully made.The capacity of the library was 1700 copies.The segment size was 36kb, and the recombination fraction was 93%.Conclusion The library was the good beginning for the following steps which were gene cloning and genescreen.

Objective:To investigate the multilocus sequence typing feature of the virulence-associated genes of Staphylococcus aureus(S. aureus) separated from the clinical specimens of a multi-center cohort children in Guangzhou area. Methods:A total number of 412 Staphylococcus aureus strains isolated from 2 059 non-repeated fecal specimens of children by three groups′ researchers in Guangzhou Women and Children′s Medical Center from August 2018 to November 2018. While collecting specimens, patient clinical information is also properly collected and preserved. After extracting the DNA of the strain, the virulence-associated genes were detected by polymerase chain reaction (PCR), including the staphylococcal enterotoxin (SE) genes ( sea, seb, sec, sed, see) and the Panton-Valentine leucocidin-encoding gene ( pvl).The multi-locus sequence typing (MLST) method was performed to reveal the MLST feature of these genes and the statistical difference were examined by the the χ 2 test. Results:Among the 412 isolates of S. aureus, 256 strains (256/412, 62.1%) contains at least one SE gene. Among the enterotoxin gens, the sec (125/412, 30.3%), seb(98/412, 23.8%)and sea (66/412, 16.0%)genes were the three most prevalent members of SEs. The frequency of pvl gene in Staphylococcus aureus was 18.7%(77/412).Among them, the frequency of Staphylococcus aureus sea gene isolated from patients with gastroenteritis (58/319, 18.2%) was significantly higher than that from the non-gastroenteritis group (8/93, 8.6%)(χ2=4.912, P=0.027). The frequency of Staphylococcus aureus pvl gene isolated from the patients with pneumonia (8/21, 38.1%) was greater than that from the non-pneumonia group (6/47, 12.8%)(χ2=4.252, P=0.039). In addition, the virulence-associated gene of S. aureus was closely related to the specific ST type, 82.4% (28/34) of ST6 carried sea gene, all ST338 and ST59 carried seb gene, 96% (48/50) ST45 carried sec gene, and the pvl gene carrying rate of ST338 was 5/5. Conclusions:The SEA toxin produced by ST6 Staphylococcus aureus may be closely related to the diagnosis of gastroenteritis in children. The frequency of pvl virulence gene in Staphylococcus aureus in children with community-acquired pneumonia was higher than that in the non-pneumonia group, and closely related to the CC59.

Objective:To retrospectively assess the relationship between immune disorder and acute gastrointestinal injury (AGI) in patients after severe polytrauma.Methods:Totally 205 patients with severe polytrauma admitted to Tongji Hospital from April 2018 to October 2019 were enrolled as the observation group, and 23 healthy volunteers were served as the control group. According to the diagnostic criteria of AGI, all patients were divided into the AGI group (with AGI) or N-AGI group (without AGI), AGI patients were divided into the S-AGI group or L-AGI group according to the severity. The levels of cytokines and lymphocyte subset were evaluated at day 1, 7, and 14 after severe polytrauma. The differences between groups were statistically analyzed. The independent risk factors of AGI were analyzed by Logistic regression analyzed.Results:Totally 79.5% (163/205) of patients with severe polytrauma were accompanied by AGI. There were significant differences in the ratio of Tc, Th at day 1 after trauma, the levels of IL-6, TNF-α, IL-8, IL-10, the ratio of Ts, Th/Ts, Treg at day 7 after trauma, and the levels of IL-8, IL-10,the ratio of Ts, Th/Ts, Treg at day 14 after trauma between the AGI group and N-AGI group ( P<0.05). There were significant differences in the ratio of Tc, Th, the levels of IL-6, TNF-α at day 1 after trauma and the ratio of Ts, Th/Ts, Treg, the levels of IL-8, IL-10 at day 7 and 14 after trauma between the S-AGI group and L-AGI group ( P<0.05). Logistic regression analysis showed that Ts 7 d ( OR=2.018, 95% CI: 1.105-5.364, P=0.013), Treg 14 d ( OR=3.612, 95% CI: 1.375-8.476, P=0.006), IL-6 7 d ( OR=1.824, 95% CI: 1.011-5.835, P=0.024), IL-10 14 d ( OR=2.847, 95% CI: 1.241-6.216, P=0.014), TNF-α 7 d ( OR=1.754, 95% CI: 1.215-5.441, P=0.018) were independent risk factors in patients with AGI after severe polytrauma. Conclusions:AGI is more easily occurred in patients with the heavier immune disorders after severe polytrauma. AGI can also aggravate pre-existing immune disorders in patients after severe polytrauma.