Analysis of phenylalanine hydroxylase gene mutations in Han ethnic children with phenylketonuria of Xinjiang region / 临床检验杂志

Objective To investigate the characteristics of phenylalanine hydroxylase (PAH) gene mutations in Han ethnic children with phenylketonuria of Xinjiang region.Methods The mutations in the promoter,exons 1-13 and flanking introns of PAH genes from 71 Han ethnic PKU children and their parents of Xinjiang region were detected by PCR,DNA sequencing and high-throughput targeted sequencing,and the obtained results were compared with those from other four provinces in northwest of China,Japan and Europe.Re suits A total of 37 kinds of mutations,including missense mutation,splice site mutation,nonsense mutation,deletion mutation and frameshift mutation,were detected in 90.1％ (128/142) of PAH alleles from 71 Han ethnic PKU children of Xinjiang region.Most mutations existed in exons 7,6,3,12,2 and 11 and intron 4 of PAH gene.The most common missense mutations were R243Q (21.8％) and R53H (7.7％).The most common splicing sites were EX6-96A ＞ G(6.3％),IVS4-1G ＞ A(4.9％) and V399V (4.2％).Moreover,The most common nonsense mutations were R111X(4.9％) and Y356X(4.9％).The detection rate of R53H mutation (7.7％) in Han ethnic PKU children of Xinjiang region was significantly higher than that in other 4 provinces of northwest of China,and a novel PAH gene nutation P225S(c.673C ＞ T) was found.Conclusion The mutation spectrum of PAH gene in Han ethnic PKU children of Xinjiang region is similar to that in other 4 provinces of northwest of China,but significantly different from that of Japanese and European population,which displays a distinct and conservative characteristic.

Effects of ultimodal nalgesia on postoperative pain and postoperative cognitive function in elderly patients undergoing esophageal cancer / 临床麻醉学杂志

Objective To investigate the effects of a variety of different methods of analgesia on postoperative pain and cognitive function in elderly esophageal cancer patients.Methods Sixty elderly pa-tients scheduled for the left into the thoracic esophageal cancer surgery were randomly divided into two groups (n =30).Group A:Before the closure of thoracic cavity to block intercostal nerve with 0.375% rop-ivacaine,followed by intravenous pumps for analgesia,formulation of sufentanil 3 μg/kg+flurbiprofen 100 mg,pump speed 2 ml/h,self-controlled analgesia 0.5 ml/pressing,locking time 15 min.Group B:Before the closure of thoracic cavity given sufentanil 10 μg+flurbiprofen 50 mg as loading dose followed by epidural analgesia pump,recipe with group A.Two groups were observed mini mental state examination (MMSE) score 1 d before surgery and 3,5,7 d after surgery,each time point visual analogue pain score (resting and exercise VAS)score postoperative within 48 h,BCS comfort score and effective pressing times of postopera-tive analgesia pump.Results Compared with group B,the rest and exercise VAS scores of group A at post-operative recovery,4,8,12,24,48 h were significantly lower (P <0.05);the BCS scores of group A at postoperative 4,8,12,24,48 h were significantly higher (P <0.05);the pressing times of group A at postoperative 4,8,12,24,48 h were significantly reduced (P <0.05);the MMSE scores of group A at postoperative 3,5,7 d were significantly higher (P <0.05);the incidence of POCD of group A on postop-erative 3,5,7 d were significantly lower.Conclusion Thoracic surgery perioperative multimodal analgesia (intercostal nerve block and intravenous analgesia)can relieve postoperative pain,reduce the incidence of POCD,improve the postoperative patient comfort and help postoperative patients with rapid recovery.

Effects of pretreatment with different doses of phosphocreatine on hepatic ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To evaluate the effects of pretreatment with different doses of phosphocreatine on hepatic ischemia-repeffusion (I/R) injury in rats.Methods.Thirty male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 5 groups (n =6 each):sham operation group (group S),hepatic I/R group (group I/R),and pretreatment with different doses of phosphocreatine groups (groups P1-3).Hepatic I/R was induced by 90 min occlusion of the hepatic artery and portal vein entering the middle and left lobes of the liver followed by 4 h reperfusion in anesthetized rats.Phosphocreatine 50,150 and 450 mg/kg were injected via the tail vein at 60 min before ischemia in groups P1-3,respectively.In groups S and I/R,the equal volume of normal saline was given instead.Blood samples were taken from the abdominal aorta at 4 h of reperfusion for determination of plasma alanine aminotransferase (ALT),aspartate aminotransferase (AST),TNF-α and IL-1β concentrations.The rats were then sacrificed and the livers were removed for determination of myeloperoxidase (MPO) activity (by ELISA),intercellular adhesion molecule-1 (ICAM-1) expression (by immunohistochemistry),and cell apoptosis (by TUNEL) and for microscopic examination (by electron microscopy).Results The MPO activity in liver tissues,plasma ALT and AST activities,TNF-α and IL-1β concentrations and the number of apoptotic cells were significantly higher in groups I/R and P1-3 than in group S,while lower in groups P1-3 than in group I/R (P ＜ 0.05).The parameters mentioned above were decreasedin turn in groups P1-3 (P ＜ 0.05).Conclusion Phosphocreatine pretreatment can attenuate the hepatic I/R injury in rats in a dose-dependent manner and inhibition of the inflammatory responses is involved in the mechanism.

Study on the T cells of T cell receptors BV complementarity determining region 3 lineage polymorphism with peripheral blood in ankylosing spondylitis patients / 中华风湿病学杂志

Objective To study the T cells lineage polymorphism of TCR BV CDR3 in the peripheral blood of ankylosing spondylitis (AS) patients,in order to provide experimental basis for the immunological patho-genesis study of AS.Methods Twenty-six subfamilies of CDR3 T cells of TCR BV in the PBMC of AS patients were amplified by RT-PCR method,then TCR BV CDR3 lineages polymorphism were analyzed by immunization scanning spectrum.Results TCR BV CDR3 scanning spectrum of 20 active AS patients showed abnormal distribution peak,including monoclonal,oligoclonal/oligoclonal trend,skewing peak and irregular abnormal peak.Among them,some subfamilies of 18 patients showed oligoclonal/oligoclonal trend expansion,BV16 and BV18 two subfamilies of one case showed monoclonal expansion.Most spectral type of PBMC TCR BV CDR3 in five normal controls showed Gauss distribution.Conclusion TCR BV CDR3 lineage have significant characteristic polymorphism and spectrum drift characteristics in the peripheral blood of AS patients,which further indicate that T cells has plaied an important role in the immunological pathogenesis of AS.Monoclonal/oligoclonal expansion of T cells may be autoreactive T cells in nature and they may be involved in the pathogenesis of AS.

Analysis of birth defects in Xinjiang multi-ethnic region / 中国组织工程研究

BACKGROUND: Xinjiang is a multi-ethnic region with significant differences in local geographical position, economic development and climatic environment. OBJECTIVE: To analyze the occurrence and development tendency of birth defects, disease categories and disparity among different ethnic groups and regions in Xinjiang.METHODS: A stratified cluster random sampling observation was performed in 13 counties (cities) according to the status of ethnical distribution and local economics of Xinjiang. Quarter Report Sheet on Babies and The defect babies register card were filled as the scheme of Chinese birth defect monitoring, and ICD10 diagnostic code was adopted in birth defect diagnosis. The birth defects rate was calculated from January 2005 to December 2008, and the disease categories and disparity among different ethnic groups and regions in Xinjiang were analyzed. RESULTS AND CONCLUSION: The average incidence rate of birth defect was 9.74‰, which was dramatically descended in 2006 and ascended afterward yearly. The incidence rate of countryside was higher than city, and male more than female. In geography, south of Tianshan Mountain was higher than north and east in birth defect incidence. Among major ethnic groups in Xinjiang, Sibe and Uygur had the highest birth defect incidence rate, followed by Man, Hazakh, and Han. The birth defect incidence of Han, Uygur and Hazakh people showed descend tendency, Hui, Mongolia, and Man people fluctuated, yet Sibe's rate had a change of rise and fall. The first five birth defect entities were neural tube deformity, cleft lips, anencephaly, congenital hydrocephalus and cleft palate combined with cleft lips. The birth defects rates are different from ethnic groups and regions in Xinjiang.

Identification of four novel mutations in the phenylalanine hydroxylase gene in patients with PKU / 中华检验医学杂志

Objective To investigate the distributions of PAH gene mutation and provide guidance for gene diagnosis and prenatal diagnosis of patients with PKU in Xinjiang of China.Methods A total of 15 patients (aged from 2 to 10 years, all with blood Phe concentration over 700 μmol/L) who visited Urumqi general hospital of Lanzhou Command were clinically diagnosed as PKU and were included in this study. PCR followed by DNA sequencing was performed to analyze the promoters, all the 13 exons and their flanking introns of PAH gene in these 15 PKU patients.Results PAH gene of 15 PKU patients was amplified by PCR, and PCR products were subjected to DNA sequencing directly.Four PAH gene mutation types, including 5′- Flanking-626G > A, 5′-Flanking-480DelACT, S196fsX4 and IVS8+1G>C, were identified in each of four PKU patients.Consequently reverse DNA sequencing showed G>A at -626 site, ACT deletion at -480 position in the promoter of PAH gene, an insertion at 584 site in the coding region and G>C at the border between exon 8 and intron 8 of PAH gene, respectively. After inquirying from PAH website and international PAH database (www.pahdb.mcgill.ca), these four PAH gene mutation types were verified as novel PAH gene mutations. Additionally, four patients carrying either of these four PAH gene mutation aged 3-5 years old were characterized by typical clinical phenotypes including blood Phe levels between 1 572-1 782 μmol/L, mental retardation, yellow hair and mousy odor of hair, skin and urine. Conclusions 5′-Flanking-626G>A, 5′-Flanking-480DelACT, S196fsX4 and IVS8+1G > C are identified as four novel PAH gene mutations to cause PKU directly probably either by disrupting the normal 3-D structure and affecting enzymatic activity of PAH or depressing the transcription and translation of PAH gene.Together, our identification of four novel PAH gene mutations will provide important clues for future gene diagnosis and prenatal diagnosis of PKU.

Novel tumor markers for early diagnosis: autoantibodies / 国际肿瘤学杂志

Commonly used tumor markers have poor sensitivity and specificity. A lot of research are being conducted searching for the ideal novel serum markers. There are a large number of tumor-associated autoantibodies in cancer patients's serum. Screening and detecting autoantibodies may be used in the early diagnosis of cancer.

Potential role of multiple tumor markers chip in preoperative diagnosis of gastric cancer / 国际肿瘤学杂志

Objective To explore the potential value of multiple tumor markers chip( C- 12) in preoperative diagnosis of gastric cancer. Methods The serum levels of 12 rumor markers were measured in 45 gastric cancer patients, 38 benign gastrosis patients and 65 normal controls by use of C-12 in order to find out the most levels of CA199, CEA, CA242, AFP and CA125 in the gastric cancer patients were significantly higher than those of the benign gastrosis patients and normal controls. Moreover, the serum levels of β- HCG and HGH were also significantly higher in gastric cancer group than benign gastric disease group and control group ( P ＜sis of gastric cancer. CEA is the TM with the highest sensitivity, validity and negative predictive value of 57.8% ,81.8% and 77.1% ,respectively whereas CA242 is the TM with the highest specificity and positive CEA + CA125 + CA199 and CEA + CA242 + CA199 + CA125, respectively. The sensitivity, specificity and validity of the best combination of 2 TMs, 3 TMs and 4TMs for gastric cancer were not statistically significantly different from those of C-12 and the best TM ( P ＞ 0.05 ). Conclusion The multiple tumor markers chip ( C-12 ) has a relatively high value in the preoperative diagnosis of gastric cancer. The best combinations of 2 TMs ( CEA + CA125) ,3 TMs ( CEA + CA125 + CA199 ) and 4TMs ( CEA + CA242 + CA199 + CA125 ) for gastric cancer diagnosis could be sufficient to replace the combination of 12 TMs.

Clinical value of multiple tumor marker protein chip detective system in diagnosis and detection of post-operative recurrence of breast cancer / 国际肿瘤学杂志

Objective To evaluate the clinical value of multiple tumor marker protein chip in diagno-sis and detection of postoperative recurrence of breast cancer.Methods The serum levels of 12 tumor makers (CA199,NSE,CEA, CA2A2,Ferritin,β-HCG,AFP,f-PSA,PSA,CA125,CA153 and HGH)were measured in 70 preoperative breast cancer patients, 32 recurrence patients,52 non-recurrence patients and 76 normal con-trois.Results ①The breast cancer group had significantly higher positive rate than that of the controls (P<0.05).The positive rates and serum levels of CA199,CEA,CA242,Ferritin,CAI25 and CA153 in breast cancer patients had those of control significant differences compared with groups (P<0.05).②The recurrence group had significantly higher positive rate than that of non-recurrence group (P<0.05).The positive rates and se-rum levels of CA199, CEA, Ferritin, CA125 and CA153 in the recurrence patients had significant differences compared with those of non-recurrence patients(P<0.05).③The positive rate of recurrence group had signif-icant difference compared with that of breast cancer group(P<0.05).Moreover,The positive rate and serum level of Ferritin in the recurrence patients had significant difference compared with that of breast cancer pa-tients.Conclusion The multiple tumor marker protein chip detective system has valid value of clinical appli-cation in the diagnosis and detection of postoperative recurrence of breast cancer.The combination detection of CA199, CEA, Ferritin ,CA125 and CA153 may be the economical and effective in the diagnosis and detection of postoperative recurrence of breast cancer.

Identification of mutations in phenylalanine hydroxylase gene of Xinjiang minority nationality phenyiketonuria patients / 中华检验医学杂志

Objective To study the mutation characteristics in phenylalanine hydroxylase gene of Xinjiang minority nationality phenylketonuria (PKU) patients and provide a scientific basis for PKU prevention and cure strategy.Methods Mutations in phenylalanine hydroxylase gene were detected by Dolymerase chain reaction-single strand comformation polymorphism (PCR/SSCP) and gene sequencing in 12 minoritv nationality patients.Results Thirteen different mutations,including 8 missense mutations,1 nonsense mutation and 3 splice mutations were found in 24 alleles.The moat common mutations were EX696A＞G and P281 L.which were respectively prevalent in Asia and Europe populations.The common mutations were R243Q,R111X,R176X and F161S.The mutation frequency of R243Q was the highest and R111X was the third highest in Northern China.R176X and F161S were two rare mutations world wide.Especially.F161S was a Chinese-specific mutation because it was for the second time that it was found in China.The mutations detected in this study were first reported in these 3 minority nationality populations,which showed a distinct ethical characteristic.Condusions There is not only a consanguineous relation but also a distinct difference in PAH gene distribution between Xinjiang minority nationality population and yellow race and Latin-American.The results suggest that Xinjiang could probably be a special PAH gene distribution region.

Detection of recombination-activating gene expression and T cell receptor gene recombination in human T cell leukemia cell lines / 中国组织工程研究

BACKGROUND: The lymphocyte-specific recombination-activating gene (RAG) 1 and 2 are essential for initiating V (D)J recombination events in both T and B cells. In addition to initiating V(D)J recombination, RAG-mediated transposition has also been proposed to contribute to chromosomal translocations and lymphoid malignancy, but the mechanism underlying this activity remains poorly understood.OBJECTIVE: To investigate RAG gene,DNA repair factors Ku70/Ku80 and terminal deoxynucleotidyl transferase(TdT)mRNA expression and T cell receptor(TCR) gene recombination in human leukemia and lymphoma cell lines.DESIGN: Repeated-determined experiment.SETTING: Institute of Molecular Immunology, College of Biotechnology,Southern Medical University.MATERIALS: Four human T leukemia and lymphoma cell lines (Jurkat,Molt-4, HuT-102, 6T-CEM) ,two Burkitt's lymphoma(Raji and Daudi) and two myelogenous leukemia cell lines (HL-60 , K-562) were cultured in complete growth medium (Hyclone,USA) , penicillin/streptomycin(50 U/mLand 50 mg/L) at 37 ℃ in a humidified atmosphere in 0.05 volume fraction of CO2. Jurkat and 6T-CEM were bought from SIBS,CAS(Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences). The other cell lines were conserved by Institute of Molecular Immunology, College of Biotechnology, Southern Medical University.METHODS: The experiment was carried out in Institute of Molecular Immunology, College of Biotechnology, Southern Medical University from October 2005 to January 2006.Reverse transcription-polymerase chain reaction (RT-PCR)was performed to examine the expression of RAG1,RAG2,terminal deoxynucleotidyl transferase(TdT), Ku70 and Ku80 genes,reparative factors,in pathway of non-homologous end joining (NHEJ);Nested and semi-nested PCR reactions were performed with locus-specific primer sets to determine TCR Dβ-Jβ signal joint T-cell receptor excision DNA circles (sjTRECs); Ligation-mediated PCR(LM-PCR) assay was used to detect the recombinational intermediates ds RSS breaks of TCRβ locus with primers specific for the flanking sequences of both Dβ1 and Dβ2.Thus,gene expression during initiating V (D)J recombination and generation of TCR gene recombination intermedium could be known.MAIN OUTCOME MEASURES: RAG gene,DNA repair factors Ku70/Ku80 and TdT mRNA expression and TCR gene recombination in human leukemia and lymphoma cell lines.RESULTS: Determination results by RT-PCR showed that RAG1 was expressed in four T cell lines and was not expressed in two Burkitt's lymphoma and two myelogenous leukemia cell lines. RAG2 and TdT were only expressed in Jurkat,Molt-4 and 6T-CEM, but TdT expression in 6T-CEM was very low. Except that Ku80 expression was not detectable in HL-60,Ku70 and Ku80 were expressed in all the cell lines. However, Determination results of TCR gene in T cell lines recombination intermedium showed that the TCR gene recombination process was not occuring in all the RAG-expression T cell lines. Ongoing TCR gene recombination was only found in Jurkat, which represent a mature stage of T cell development. In addition,characteristic junctional diversity of signal joints was observed in DNA isoated from Jurkat.CONCLUSION: T leukemia and lymphoma may be more likely to have a relation with RAG.The results give a clue that Jurkat can probably provide an ideal cell line modle for studying RAG and T cell lymphomagenesis.