Objective:To explore whether endoscopic surgery can achieve the same effect on thyroid cancer as open surgery.Method:44 cases were selected to endoscopic thyroidectomy with breast areola approach, who were prepared to accept the surgical treatment of thyroid cancer with cT1N0 stages, female, ≤60 years old, no history of neck surgery and beauty desire. Fifty patients with the same conditions undergoing thyroid open surgery were as the control group.Results:Operative time of endoscopic group[ (170.0±28.0) min] was longer than that of the open operation group[ (90.0±21.0) min ( t=15.610, P=0.000) ]. There was no significant difference between the two groups in surgical complications ( P>0.05) . There was no statistical difference between the number of paratracheal and anterior tracheal lymph nodes of endoscopic group (6.0 ±4.2) and open surgery group (5.5 ±3.7) ( t=0.692, P>0.05) . There was no statistical difference between the number of anterior laryngeal lymph nodes of endoscopic group (0.7 ±1.1) and open surgery group (0.5 ±0.9) ( t=1.186, P>0.05) . Conclusion:The breast areola approach endoscopic thyroidectomy is safe and reliable in treatment of thyroid cancer, and the central cervical lymph node dissection is sufficient, which can be used as the choice of operation mode for thyroid cancer patients in cT1N0 stage.

Objective To study the impact of splenectomy and esophagogastric devascularization on the nutritional status of patients with cirrhosis and portal hypertension.Methods Sixty consecutive patients with cirrhosis and portal hypertension who underwent splenectomy and esophagogastric devascularization at the Beijing YouAn Hospital from April 5,2015 to January 23,2017 were included in this study.The body mass index (BMI),albumin (Alb),prealbumin (PA) and lymphocyte counts were prospectively collected at the end of 1-week,1-month,3-month,6-month and 1-year after surgery.The postoperative results were compared with the preoperative results in these patients.Results The BMI results obtained at 1-week and 1-month after surgery were significantly lower than the preoperative level [(22.14 ± 3.08)kg/m2 vs.(22.85 ± 3.14) kg/m2,(21.72 ± 3.05) kg/m2 vs.(22.86 ± 3.16) kg/m2,P ＜ 0.05].The BMI result at the end of 1-year after surgery was significantly elevated when compared with the preoperative level [(23.24 ± 3.64) kg/m2 vs.(22.68 ± 3.47) kg/m2,P ＜ 0.05].The ALB levels at 1-month and 3-month after surgery were significantly higher than the preoperative level [(39.87 ± 4.22)g/L vs.(35.35 ±5.15) g/L,(39.35 ± 4.75) g/L vs.(34.82 ± 5.50) g/L,P ＜ 0.05].The PA obtained at 1-week after surgery was significantly lower than the preoperative levels [(79.59 26.52)mg/L vs.(121.77 ±39.96)mg/L,P ＜ 0.05].The lymphocyte counts at all the points after surgery were significantly higher than the preoperative level (P ＜ 0.05).Conclusion Short term and long term nutritional status improved in patients with cirrhosis and portal hypertension after splenectomy and esophagogastric devascularization.

BACKGROUND:As the sequencing technology has been widely used and high-resolution confirmation of organ transplant matching has been gradualy developed, new human leukocyte antigen (HLA) aleles are emerging. However, the gene frequency of some genes cannot be calculated accurately, and there are rare reports. These genes are often ignored, and it is easy to misjudge their genotypes only according to gene frequency. OBJECTIVE:To test and analyze a rare alele, HLA-C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. METHODS: Genomic DNA was extracted automaticaly from the blood sample by using quick DNA purified kit and amplified by HLA-C locus commercial sequence-based typing kit. The purified PCR product was utilized as the DNA template in the sequencing reaction, and six direct sequencing reactions of PCR product covering exons 2, 3 and 4 in both directions were performed using commercial kit. Four direct sequencing reactions of PCR product covering exon 5 in both directions, exon 6 in forward direction and exon 7 in reverse direction were performed using in-house BigDye terminator cycle sequencing reaction kit. Sequencing reaction products purified by ethanol/sodium acetate/ ethylenediaminetetraacetic acid method were sequenced by ABI PrismTM3730 DNA Sequencer. RESULTS AND CONCLUSION:The alele assignment was analyzed with Assign-SBT 3.6+ software, and the sample HLA-C typing result was C*07:04, 08:99. Increasing the sequencing analysis at exons 5, 6 and 7 of HLA-C locus wil help to make clear the ambiguous SBT result and improve the accuracy of HLA-C typing when it is necessary, which shows important significance in clinical tissue matching. Cite this article:Wang DM, Zou HY, Nie DM, Gao SQ, Wang F.Sequencing analysis of a rare human leukocyte antigen, C*08:99, from a volunteer donor of hematopoietic stem cel transplantation. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):102-106.

OBJECTIVETo investigate the number and ratio of ambiguous allele combinations from human leukocyte antigen (HLA) confirmatory test by sequencing based typing for unrelated donor marrow transplantation, and to establish an efficient strategy for identifying such ambiguities.

METHODSA total of 650 donor-receipt samples were genotyped for 5 loci of the HLA gene using an Atria SBT commercial kit. Exons 2, 3 and 4 of HLA-A, -B and -C, exon 2 of HLA-DRB1 and exons 2 and 3 of HLA-DQB1 were tested by routine HLA genotyping. The ratio of usual ambiguous allele combination was calculated. The ambiguities were subjected to further confirmatory test by PCR-SSP or PCR-SBT retest at outside of the routine sequencing region.

RESULTSAmong the 650 tested samples, the ratio of ambiguity at HLA-A, B, C, DRB1 and DQB1 were 76.31% (496/650), 91.08% (592/650), 97.69% (635/650), 88.62% (576/650) and 43.38% (141/650), respectively. A total of 36 ambiguous allele combinations inside the routine sequencing region and 22 ambiguous allele combinations outside of the routine sequencing region were discovered. After removing rare alleles based on the Chinese common and well documented (CWD) Allele Table (Version 1.01), 9 ambiguous CWD allele combinations inside the routine sequencing region, including 3 located in HLA-B, HLA-C and 1 located in other three HLA loci were found. Ten ambiguous CWD allele combinations outside of the routine sequencing region, including 4 located in HLA-C, -DRB1 and 1 in HLA-A, -B respectively were determined. All samples with ambiguous CWD allele combinations could be distinguished by high-resolution PCR-SSP commercial kits or PCR SBT retest at outside of the routine sequencing region.

CONCLUSIONThe common and well documented allele combinations in sequencing-based typing at five HLA loci have been analyzed. Our strategy may provide valuable information for more efficient, low cost and accurate method for high resolution genotyping of HLA genes.

Alleles ; Genotype ; HLA Antigens ; genetics ; Humans

Objective To investigate the role of cytokines combined with CLIF Consortium organ failure score (CLIF-COFs) in determining the prognosis of liver transplant in hepatitis B-related acute-on-chronic liver failure (HB-ACLF) patients.Methods Thirty-seven cases of HB-ACLF patients undergoing liver transplantation were divided into HB-ACLF patients with complications group (n =15) and those without complications (n =22).Plasma were prospectively collected immediately before LT and on the 1st,3rd,5th,7th day after LT in HB-ACLF patients.The serum levels of twenty-seven cytokines were determined by 200 LUMINEX liquid chip technology.Cytokines and CLIF-COFs were analyzed with logistic regression and the receiver operating characteristic to confirm the correlation with the total complications post-LT.Results The serum levels of post-transplant G-CSF and MCP-1 in HB-ACLF patients with complications were higher than those of without complications (P ＜ 0.05).The COX analysis indicated that MCP-1 and CLIF-COFs were predictors of post-LT complications [A UC:0.821,95％ CI:0.668-0.974;AUC:0.738,95％ CI:0.578-0.898].The discriminatory power of MCP-combined with CLIF-COFs [AUC:0.839,95％ CI:0.703-0.975] was better than that of either.Conclusions MCP-1 combined with CLIF organ failure score can better predict the short-term outcomes of liver transplantation in HB-ACLF patients.

OBJECTIVETo investigate the genetic basis for a novel allele HLA-C*01:78.

METHODSGenomic DNA was extracted from peripheral blood using a QIAGEN quick DNA extraction kit. The regions encompassing HLA-C from exon 1 to intron 3 and intron 3 to 3'UTR were amplified and cloned using a cloning sequencing kit in order to split the two alleles apart. Selected clones were sequenced to include exons 2 to 4.

RESULTSSequencing results have indicated the HLA-C alleles of the proband to be a novel C*03:04 allele. The sequence has been submitted to GenBank (KF049216). BLAST analysis has confirmed the novel allele to have one nucleotide difference as C*01:03 at genomic nt316C>A (codon 82CGC>AGC) in exon 2, which has resulted in replacement of one amino acid (82R>S).

CONCLUSIONThe novel allele has been officially named as C*01:78 by the WHO Nomenclature Committee. The HLA allele type of the proband was therefore A*02:07, 24:02; B*40:01, 46:01; C*01:78, 03:04; DQB1*05:02, 05:02; DRB1*16:02, 16:02.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Exons ; Female ; HLA-C Antigens ; genetics ; Humans ; Introns ; Leukemia ; genetics ; Male ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo explore the reason for HLA-DQB1 allele dropout during routine sequence-based typing(SBT) in order to improve the accuracy of typing.

METHODSTwo thousand samples derived from HLA high-resolution typing laboratory were typed for HLA-DQB1 locus using an AlleleSEQR HLA-DQB1 SBT kit. Non-conclusive results and "abnormal" sequencing samples were retyped using a LABType rSSO HD HLA-DQB1 kit and further analyzed with both sequence-specific primers and group-specific primers and sequenced for haplotype analysis.

RESULTSAmong the 2000 samples, 2 samples with no conclusive result were identified. The heterozygosity was confirmed with both the LAB Type SSO HD HLA-DQB1 kit and PCR-SBT in house method. Subsequent HLA-DQB1 cloning and haplotype sequencing have elucidated that HLA-DQB1*02:02 dropped out at exon 2 for the first sample and HLA-DQB1*02:01:01 dropped out at exon 2 for the second sample during PCR amplification. No novel nucleotide mutation was found.

CONCLUSIONOur results indicated that preferential amplification at exon 2 of DQB1 may result in allele dropout in exon 2 sequences during HLA-DQB1 SBT test. This may provide useful information for HLA genotyping.

Alleles ; DNA Primers ; genetics ; Exons ; Genotype ; HLA-DQ beta-Chains ; genetics ; Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods

BACKGROUND:Many articles concerned novel al eles reported in China and outside China, but some al eles were detected lately. After that, these al eles were tested again. Because frequencies of these al eles are very low, few relevant articles are reported, so these al eles are ignored easily. OBJECTIVE:To test and identify four human leukocyte antigen rare al eles that patients and donors carries. METHODS:Genomic DNA was extracted automatical y from blood samples using quick DNA purified kit, typed by human leukocyte antigen locus commercial sequence-based typing kits and confirmed by sequence-specific primers high-resolution kits. RESULTS AND CONCLUSION:Four detected rare al eles are B*27:15, B*51:39, C*07:66 and C*08:22. The four above-mentioned human leukocyte antigen rare al eles defined by National Marrow Donor Program are not rare in China. The facts prove that C*08:22 which was defined a rare al ele by National Marrow Donor Program before is a common al ele in Chinese Han nationality.

Objective To analyze the management and outcomes of patients with ST-segment elevation myocardial infarction (STEMI) in Liaoning province.Methods The data were collected from a prospective and multicenter registry study including 8 tertiary hospitals and 12 secondary hospitals in Liaoning province.Total 1429 patients with acute STEMI admitted to hospitals from June 2009 to June 2010 were included in the study.A unified follow-up questionnaire was applied on patient discharged.Results The average age of patients was (63 ± 13)years.37.4％ of patients recognized the disease as heart disease and 39.7％ were transported by emergency ambulance with a median symptom-to-door time of 150 min.52.9％ patients underwent emergency reperfusion therapy,including fibrinolytic therapy (24.4％) and primary percutaneous coronary intervention (PCI,28.1％).The in-hospital treatment included aspirin (99.6％),clopidogrel (81.9％),statins (90.1％),low molecular weight heparin (89.5％),β-blocker (66.0％),angiotensin converting enzyme inhibitor (ACEI)/ angiotensin receptor blocker (ARB)(66.6％).The in-hospital mortality was 10.7％ ; the mortality in females was higher than that in males (18.3％ vs.7.9％,P ＜ 0.01) and the mortality in older patients (≥ 65 years) was higher than that in younger patients (＜65 years)(17.0％ vs.5.2％,P ＜0.01).The follow-up treatment included:aspirin (81.1％),clopidogrel (45.0％),statins (61.0％),β-blocker (48.3％),ACEI/ARB (42.4％).The follow-up mortality was 5.0％ after hospital discharge.Conclusions Longer pre-hospital delay is commonly seen in STEMI patients.There is still certain gap of emergency reperfusion therapy and the evidence-based medication with related clinical guidelines of STEMI management in Liaoning.

The Rictor/mTOR complex plays a pivotal role in a variety of cellular functions including cellular metabolism, cell proliferation and survival by phosphorylating Akt at Ser473 to fully activate the Akt kinase. However, its upstream regulatory pathways as well as whether it has additional function(s) remain largely unknown. We recently reported that Rictor contains a novel ubiquitin E3 ligase activity by forming a novel complex with Cullin-1, but not with other Cullin family members. Furthermore, we identified SGK1 as its downstream target. Interestingly, Rictor, but not Raptor or mTOR, promotes SGK1 ubiquitination. As a result, SGK1 expression is elevated in Rictor(-/-) MEFs. We further defined that as a feedback mechanism, Rictor can be phosphorylated by multiple AGC family kinases including Akt, S6K and SGK1. Phosphorylation of Rictor at the Thr1135 site did not affect its kinase activity towards phosphorylating its conventional substrates including Akt and SGK1. On the other hand, it disrupted the interaction between Rictor and Cullin-1. Consequently, T1135E Rictor was defective in promoting SGK1 ubiquitination and destruction. This finding further expands our knowledge of Rictor's function. Furthermore, our work also illustrates that Rictor E3 ligase activity could be governed by specific signaling kinase cascades, and that misregulation of this process might contribute to SGK overexpression which is frequently observed in various types of cancers.
Animals ; Carrier Proteins ; metabolism ; Cell Proliferation ; Cells, Cultured ; Cullin Proteins ; genetics ; metabolism ; Fibroblasts ; metabolism ; Humans ; Immediate-Early Proteins ; metabolism ; Mice ; Phosphorylation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rapamycin-Insensitive Companion of mTOR Protein ; TOR Serine-Threonine Kinases ; genetics ; metabolism ; Ubiquitin ; genetics ; metabolism ; Ubiquitination