BACKGROUND:It has great significance to understand individual human leucocyte antigen allele(HLA) haplotype and genotype in organ allotransplantation.OBJECTIVE:To analyse HLA allele frequency,haplotype frequency and characteristices in 36 cases of living-related donor kidney transplantation.DESIGN,TIME AND SETTING:The gene polymorphism analysis was performed in the Tissue Typing Laboratory of Organ Transplantation of Ruikang Hospital of Guangxi Traditional Chinese Medical University from January 2007 to June 2008.PARTICIPANTS:Thirty-six cases of living-related donor kidney transplantation,who were relatives of Guangxi Han population.METHODS:HLA-Ⅰantigen was detected by dry plates of monoclonal antibodies,and HLA-Ⅱ antigen was subjected to genotyping using PCR-SSP technique.And then the allele and haplotype frequencies of HLA-A,B,DRB1 were calculated.MAIN OUTCOME MEASURES:HLA haplotype and genotype of Guangxi Han population.RESULTS:A total of 11 HLA-A,24 HLA-B and 13 HLA-DR alleles were detected among 36 cases,showing a rich polymorphism.A2-B46,A11-B60,B46-DRB1*09,B60-DRB1*15 and A2-B46-DRB1*09 among 144 haplotypes were informative with frequency higher than 0.05.A total of 10 HLA-A-B haplotypes and 9 HLA-B-DRB1 haplotypes were in strongest linkage disequilibrium.Distribution of HLA-A,B,DRB1 alleles and haplotypes in Guanxi population is similar to Han population in South China,such as Hunan and Taiwan,but had its own characteristics.CONCLUSION:HLA alleles have richer polymorphisms with exhibit geographic genetic characteristics.

Objective To investigate whether the protein kinase C inhibitor can promote the apopto- sis of multidrug resistance tumor cell lines which are induced by chemotherapy drugs.Methods Choose the KB/S(oral squamous cancer cell line)and KB/VCR(its multidrug resistant cell line)to compare the Adri- amycin-induced apoptosis with or without staurospolin(protein kinase C inhibitor).The apoptosis is stained with acridine orange,tested by flow cytometry,and approved by electron microscope.Results 36 hours after the treatment with 0.04 ?g/ml adriamycin,apoptotic cells of KB/S are 96.68%,and after 48 hours,the apop- totic cells of KB/VCR are 64.99%.When the concentration of adriamycin are augmented to 0.4?g/ml and 2.0?g/ml,the apoptotic cells of KB/VCR are 69.74% and 37.18% respectively.When treated with stau- rospolin together,the apoptotic cells of KB/VCR increased to 72.58%(?~2=4.5,P0.05)respectively.These results were testified by electron microscope and acridine orange-stain.Conclu- sion The resistance to apoptosis may be one of the mechanisms of multidrug resistance and the protein ki- nase C inhibitor may reverse this resistance by promoting the apoptosis of multidrug resistance tumor cells.

Objective To examine the suscepribility of the single nucleotide polymorphisms(SNPs)in IL-1 gene in Chinese Han population to ankylosing spondylitis (AS). Methods An correlation analysis was performed in a case-control cohort of 162 AS cases, 58 patients with other autoimmune diseases and 162 controls. Four SNPs located in the IL-1 gene (rs16944, rs3811058, rs419598, rs315952) were examined by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP). Results The frequencies of allele C at position rs16944, rs3811058, rs419598 significantly increased in AS cases VS controls, so did their genotype frequencies (50% vs 36.3%, 57.9% vs 52.8%, 45.7% vs 12.3%, P＜0.05). The SNPs of these sites significantly influenced the prevalence of AS. Conclusion We discover that SNPs of IL-1 gene Rs16944, Rs419598, Rs3811058 is closely related to the occurrence of AS.

BACKGROUND: The quantity and bioactivity of isolated islet are vital to islet transplantation; while, the cold ischemia time and human leucocyte antigen (HLA) typing are key factors to islet quantity and bioactivity which inflect islet transplantation.OBJECTIVE: To observe the effects of cold ischemia time and blood compatibility on quantity and bioactivity of islet cells.DESIGN: Observational study.SETTING: Ruikang Affiliated Hospital of Guangxi College of Traditional Chinese Medicine.MATERIALS: Organs from voluntary donors died of irreversible coma were adopted whose blood-type and HLA typing had been known, pancreases acquisition was carried on after other organs ablation or in the meantime. The blood of identical ABO and HLA matching conformity, or HLA cross-matching hypersensitization (missmatching over 3 Iocuses),or panel reaction antibody (PRA) ＞ 50%, or lymphocyte cytotoxin crossmatching test positive. The isolated and purified islet suspension was filtered by 70 μm filter, which result in preparing 1.2×105/L islet suspention.METHODS: Hypertonic citrate adenine solution was perfused into aorta, and kidney-pancreases and kidney-pancreases-liver were cut together or kidney-pancreases-liver was cut separately. Islet activity was judged by diphenylthiocarbazone (DTZ) dyeing and acridine orange (AO) dyeing; meanwhile, twelve pancreases far from contamination were aquired, mean ablation time was 15 minutes; cold ischemia time ranged from 2.5 to 8 hours. Cold ischemia time of nine pancreases was controlled in 5 hours, warm ischemia time was 0-3 minutes, and peptic time was (15±2.4) minutes, correlation of islet cells and histocompatibility with survival of islet cells was analyzed.MAIN OUTCOME MEASURES: Survival rate of islet cells; counts of platelet, heterophil granulocyte and monocyte.RESULTS: The cutting of kidney, pancreases and liver were successful. If the cold ischemia time was controlled within 5 hours, activity of islets was above 80%. Pancreatic gland used for islet transplantation and cutting of other organs could not affect activity of islet cells. When human islets were exposed to human blood, it would induce a rapid consumption of platelets, neutrophils, monocytes and lymphocytes in the blood. Consumption of blood cells was more in the HLA typing groups than that in the control group. After adding heparin, there was significant difference in cell account among the three groups (P ＜ 0.05) and the reaction was relieved obviously. After 24-hour cultivation, there were significant difference in active islets quantity between HLA typing compatibility group and HLA typing incompatibility group (P ＜ 0.05).CONCLUSION: Pancreatic gland obtained under the cold ischemia time ＜ 5 hours can be used in clinical transplantation of islet cells; a good histocompatibility can raise successful rate of transplantation of islet cells.

BACKGROUND:Tumor recurrence results from the incomplete removal of cancer stem cel s with self-renewal characteristics, and then how to label and eliminate cancer stem cel s becomes the key to cancer treatment. OBJECTIVE:To observe the morphology,distribution and number of CD44+/C-myc+METHODS:Pathological tissues from 150 patients with colorectal cancer were taken to prepare tissue microarray, in order to observe and count CD44 cel s in colorectal cancer, and to explore the relationship between the expression and postoperative metastasis.+/C-myc+cel s by using immunohistochemical double staining. Patients were fol owed up through mobile phones , letters , and so on, and the relationship between the expression of CD44+/C-myc+cel s and postoperative metastasis were statistical y recorded.RESULTS AND CONCLUSION:There was no CD44+/C-myc+cel s in normal tissue and little in adenoma. A smal number of CD44+/C-myc+cel s distributed as dots or focal lesions in adenocarcinoma. The number of CD44+/C-myc+cel s was related to the degree of adenocarcinoma differentiation, depth of invasion, and lymph node metastasis (P<0.05). The univariate analysis showed that the overal survival rate and progression-free survival rate were associated with the number of CD44+/C-myc+cel s, lymph node metastasis and Ducks staging in adenocarcinoma (P<0.05). The multivariate analysis showed that the overal survival rate was related to CD44+/C-myc+cel amount and the progression-free survival rate was related to CD44+/C-myc+cel amount and the Ducks staging. Therefore, CD44+/C-myc+cel s are probably cancer stem cel s, and Ducks staging and CD44+/C-myc+cel amount are important prognostic factors for colorectal cancer.

Objective To analyze the application of modified abdomen pathway for excising the adrenal huge pheoehromocytoma.Methods One patient(male,42-year-old)had adrenal huge pheochromocytoma.The pheochromocytoma was about 15.0 cm×8.0 cm×7.0 cm.After 3 weeks' preparations,the patient was operated.The operation was made through the modified abdomen pathway for excision,without excising the transverse on peritoneal,and the interference to organs of peritoneal was reduced.The tumor on adrenal gland adhered kidney very tightly.Expanded radical excision including tumor,kidney,adrenal gland was applied.Results The operative time was 300 rain and the volume of bleeding was about 1000 ml.In the operation process,blood pressure of the patient was stable,the visual field of operation was satisfactory.Blood pressure of the patient returned normal 6 months postoperatively.And there was no indication of tumour relapse or matastasis.Conclusion The modified abdomen pathway can expose the satisfactory visual field,and is safe and effective for adrenal gland tumor operation.

Animals ; Chemical and Drug Induced Liver Injury ; drug therapy ; Liver ; drug effects ; pathology ; Mice ; Silymarin ; pharmacology ; Ursodeoxycholic Acid ; pharmacology

OBJECTIVEIsobaric tags for relative and absolute quantitation (iTRAQ) combined with mass spectrometry were used to screen differentially expressed plasma proteins in cold stress rats.

METHODSThirty health SPF Wistar rats were randomly divided into cold stress group A and control group B, then A and B were randomly divided into 3 groups (n = 5): A1, A2, A3 and B1, B2, B3. The temperature of room raising was (24.0 +/- 0.1) degrees C, and the cold stress temperature was (4.0 +/- 0.1) degrees C. The rats were treated with different temperatures until 12 h. The abdominal aortic blood was collected with heparin anticoagulation suction tube. Then, the plasma was separated for protein extraction, quantitative, enzymolysis, iTHAQ labeling, scx fractionation and mass spectrometry analysis.

RESULTSTotally, 1085 proteins were identified in the test, 39 differentially expressed proteins were screened, including 29 up-regulated proteins and 10 down-regulated proteins. Three important differentially expressed proteins related to cold stress were screened by bioinfonnatics analysis (Minor histocompatihility protein HA-1, Has-related protein Rap-1b, Integrin beta-1).

CONCLUSIONIn the experiment, the differentially expressed plasma proteins were successfully screened in cold stress rats. iTRAQ technology provided a good platform to screen protein diaguostic markers on cold stress rats, and laid a good foundation for further. study on animal cold stress mechanism.

Animals ; Blood Proteins ; chemistry ; Cold Temperature ; Mass Spectrometry ; Rats ; Rats, Wistar ; Stress, Physiological

Objective To investigate the changes of C- reactive protein( CRP ) and homocysteine ( Hcy)in the type 2 diabetes with depression,and its clinical significance and potential mechanism. Methods One hundred and twenty-four cases with type 2 diabetes were divided into the depression group(63 cases)and non-depression group( 61 cases ) according to the Self-Rating Depression Scale and verified by Self-Rating Anxiety Scale. The information including age,sex,education degree,body mass index,course of disease and the number of complications were recorded. The levels of CRP,Hcy,fasting plasma glucose( FPG ),glycosylated hemoglobin(HbA1c)and blood lipid were measured. The depression group was divided into mild,medium and heavy group to compared the changes of Hcy and CRP. Results The levels of Hcy,HbA1c and the number of complications in depression group were 11. 5( 8. 6,15. 6 )μmol/L,( 10. 13 ± 2. 17 )%,and 2( 1,3 ) respectively,higher than that of non-depression group(8. 6(7. 4,11. 2)μmol/L,(9. 33 ± 2. 20)%,1(0,2)), while the education degree of depression group((9. 75 ± 3. 36)years)was lower than that of non-depression group((11. 56 ± 3. 73)years),and the differences were significant( t/Z = -3. 537,0. 952,-2. 339,0. 228 respectively;P ﹤0. 05). The levels of Hcy in mild,medium and heavy depression group were(8. 75(7. 45, 10. 45)μmol/L,12. 2(8. 90,14. 40)μmol/L,19. 50(14. 33,28. 03)μmol/L respectively and the difference was significant(F =25. 963,P =0. 000). No significance difference was found in terms of CRP level(2. 35 (1. 10,4. 92)mg/L,3. 25(1. 11,5. 68)mg/L,2. 32(1. 27,5. 41)mg/L;F=0. 194,P=0. 907). There was significant correlation between depression scores and Hcy( r=0. 615,P=0. 000). Conclusion Type 2 diabetes with depression is associated with the level of blood glucose,education degree and the course of disease. Hcy,not CRP is an independent risk factor of type 2 diabetes with depression.

BACKGROUNDLiver transplantation has become the treatment of choice for patients with end-stage acute or chronic hepatic disease. Bile duct complications are common events after liver transplantation. The aim of this study was to evaluate the blood supply of the human bile duct and identify the underlying mechanisms of bile duct complications after liver transplantation.

METHODSThe duct supply branches from gastroduodenal artery and blood supply of extrahepatic bile duct system were re-evaluated through selective hepatic angiography from 600 patients. In addition, 33 cadavers were injected with latex casting material into the common hepatic artery, then the extrahepatic bile duct and the branches from the common hepatic artery were carefully dissected to visualize the gastroduodenal artery and its branching to the extrahepatic bile duct.

RESULTSThe bile duct artery arose from the branch of the gastroduodenal artery in 8.1% (49/600). Of these 49 individuals, the bile duct artery was supplied by the gastroduodenal artery (61.22%, 30/49), the proper hepatic artery (14.29%, 7/49), or both the gastroduodenal artery and the proper hepatic artery (24.49%, 12/49). In our study of 33 cadavers, the percentage that the bile duct artery arose from the gastroduodenal artery was 27.27%. The blood supply to the bile extrahepatic bile ducts was divided into different segments and formed longitudinal and arterial network anastomosed on the walls of the duct.

CONCLUSIONSThere is a close relationship between the duct supply branches from gastroduodenal artery and the blood supplying patterns of the extrahepatic bile duct system. In liver transplant surgery, the initial part of the gastroduodenal artery is preferred to be preserved in the donor liver. It is of great significance to improve the success rate of operation and reduce complications.

Adult ; Aged ; Aged, 80 and over ; Angiography ; Bile Ducts, Extrahepatic ; diagnostic imaging ; Female ; Hepatic Artery ; diagnostic imaging ; Humans ; Liver Transplantation ; Male ; Middle Aged