OBJECTIVE To evaluate the clinical application of T-cell based IFN-? release assays(IGRA) for the rapid diagnosis of active tuberculosis.METHODS IFN-? and HIV antibody were detected by using ELISA.Antibody to Mycobacterium tuberculosis was detected by colloidal gold.At the same time,the M.tuberculosis DNA loads were examined by FQ-PCR.Statistical analysis were performed to analyze the correlation of IFN-? with M.tuberculosis antibody and DNA,respectively.RESULTS The sensitivity of TB-IGRA was 90.24%,specificity was 93.34%;the positive rate of TB-IGRA in 82 tuberculosis patients was higher than from sputum smear(64.63%),TB-PCR(76.83%) and tuberculosis antibody(40.24%).CONCLUSIONS As a replacement of TB-PCR,IFN-? can be used as a valued index to evaluate tuberculosis infectin.

Objective To investigate the association between the apolipoprotein E (apoE) gene polymorphism and the dose for warfarin individual maintenance. Methods The genotypes of 249 patients with warfarin treatment in maintenance doses were determined by PCR/DHPLC assay. The doses for warfarin maintenance were compared among patients with different genotypes. Results In the total of 249 patients, the frequencies of 2/ε2, ε2/ε3, ε2/ε4, ε3/ε3, ε3/ε4, ε4/ε4 genotype were 1.20%, 15.66%, 1.80%, 72.29%, 9.24%, 0.80%, respectively; the allele frequencies of ε2, ε3, ε4 were 9.44%, 84.74%, 5.82%, respectively. The warfarin dose of group ε2 (ε2/ε2, ε2/ε3) was (3.24±1.36) mg/d, slightly higher than that of group ε3 (ε3/ε3, 2.91±1.14 mg/d) or group ε4 [ε4/ε4, ε3/ε4, (2.98±1.05) mg/d], but the difference of the warfarin doses among the 3 groups did not reach statistical significance (F=1.848,P＞0.05). Conclusion ApoE polymorphism may be not a major genetic factor that influences the individual dose for warfarin maintenance.

Objective:Adenoid cystic carcinoma (ACC) is an uncommon malignant neoplasm, which mostly originates from the major and minor salivary glands of the head and neck region. This study aims to provide new information on head and neck ACC with cervical lymph node metastasis. Methods:Out of the 616 patients who underwent primary tumor resection from 1995 to 2008 in the authors' hospital, 62 cases with cervical lymph node metastasis were analyzed. Results:The general incidence rate of cervical lymph node me-tastasis in ACC was approximately 10%. The base of the tongue, mobile tongue, and mouth floor were the most frequent sites of lymph node metastasis with incidence rates of 19.2%, 17.6%, and 15.3%, respectively. Most cases exhibited the classictunnel-stylemetastatic pattern of occurrence, and the levelⅠb andⅡregions were the most frequently involved areas. Primary site and lympho-vascular invasions were significantly associated with lymph node metastasis. High patient mortality rate was also significantly correlat-ed with a high number of lymph node positive cases. Conclusion:Cervical lymph node metastasis has a high tendency of occurrence in the tongue-mouth floor complex, following the classictunnel-stylemetastatic pattern. Peritumoral lymphovascular invasion could be taken as a strong predictor for the occurrence of lymph node metastasis, which ultimately leads to poor prognosis of ACC patients. A selective neck dissection should be considered as a management in such patients.

Objective To determine whether there is any difference in rocuronium-induced muscle relaxation between patients of Buyi and Han nationality.Methods Sixty ASA Ⅰ or Ⅱ patients of both sexes aged 20-55 yr,with body mass index of 20-25 kg/m2,undergoing laparoscopic or arthroscopic surgery under general anesthesia,were divided into 2 groups ( n =30 each):Han group (group H) and Buyi group (group B).Anesthesia was induced with midazolam,fentanyl and TCI of propofol (Cp=2-3 μg/ml).Tracheal intubation was facilitated with rocuronium 0.6 mg/kg.The patients were mechanically ventilated.PETCO2 was maintained at 30-35 mm Hg.Neuro-muscular (N-M) function was monitored by accelerography.N-M block was assessed by single stimulation of ulna nerve after loss of consciousness.The onset time,maximal N-M block time,clinical muscle relaxation time (from injection d rocuronium to 25％ recovery),75％ recovery time (from injection of rocuronium to 75％ recovery) and recovery index were recorded.The plasma concentration of albumin and α1-acid glycoprotein were measured by ELISA and biochemical analysis respectively.Results The onset time was significantly longer and plasma α1-acid glycoprotein concentration lower in group B than in group H.There was no significant difference in maximal N-M block time,clinical muscle relaxation time,75％ recovery time,recovery index and plasma albumin concentration between the 2 groups.Conclusion The onset time of rocuronium-induced N-M block is longer in patients of Buyi nationality as compared with patients of Han nationality.Lower plasma α1 -acid.glycoprotein concentration may be involved in the underlying mechanism.

Objective To establish an external quality assessment ( EQA) system of genitourinary tract secretions routine testing in Guizhou Province and improve the overall testing level.Methods From 2009 to 2011, more than 50 clinical laboratories in different grade hospitals from Guizhou Province were enrolled as participating units every year.EQA was carried out twice a year.Each time, five slides of high quality Wright′s or Gram stain smear of the genitourinary tract secretions or photographs obtained from these smears were selected to send to the participating laboratories for testing, and the feedback results from each laboratory were analyzed.The qualification was judged by the coincidence rate equal to or more than 80%. The average coincidence rates of each time and each year were statistically analyzed by Chi-squared test. Results From 2009 to 2011, the number of EQA participating units increased from 55 to 96, with an average return rate of >80%.Coincidence rates <80%of the 6 EQA results in the 3 years were as follow:four times for coccobacteria (73.7%,77.8%,61.1%,77.1%), twice for bacillus (75.6%,79.3%) and coccobacillus (64.3%,52.1%), once for infusorian (79.7%), epithelial cells (76.1%), neutropenia (75.7%) and cleanliness (71.3%).There were six batches of 30 quality assessment controls (accounting for 20.0%) in the six EQAs had the coincidence rate of <80%.Eleven items of 30 quality assessment controls with 1 to 15 batches were unqualified ( average coincidence rate of<80%) respectively.The item with the highest total average coincidence rate was suspected gonococcus (94.2%), and two items with the lowest total average coincidence rates were coccus and coccobacillus ( 77.0%, 75.2%, respectively ) . Conclusions This EQA program carried out within a certain range of clinical laboratories achieved good results:participating units increased significantly;the total score of all the items showed an obviously upward trend;the quality awareness of clinical lab technicians has enhanced to a certain extent.In this study, EQA system of genitourinary tract secretion routine testing were preliminarily established in Guizhou province, which provided a reference model of internal quality control ( IQC ) and EQA for clinical laboratories and higher authorities, and will be bound to have a positive impact on improvement of the overall level of genitourinary tract secretion routine testing.

OBJECTIVE: To detect mutation of LBR gene in a pedigree affected with Pelger-Huёt anomaly (PHA) and to explore its clinical characteristics. METHODS: Genomic DNA was extracted from the pedigree and healthy controls. The 14 exons of the LBR gene were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified in other family members and 100 healthy controls. Polyphen-2 and SIFT software were used to predict the effect of the mutation, and Swiss-model software was used to simulate the protein structure. RESULTS: Three patients were found to carry a c.893G>A mutation in exon 8 of the LBR gene, which resulted in substitution of the 298th amino acid residue glycine by glutamic acid (p.Gly298Glu). The same mutation was not found in healthy family members and 100 healthy controls. The mutation was predicted to be damaging. Bioinformatic simulation showed the mutation has altered the 3D structure of the LBR protein. CONCLUSION The c.893G>A (p.Gly298Glu) mutation in the LBR gene probably underlies the PHA in this pedigree and has enriched the spectrum of LBR gene mutations.
Case-Control Studies ; DNA Mutational Analysis ; Exons ; Humans ; Mutation ; Pedigree ; Pelger-Huet Anomaly ; genetics ; Polymerase Chain Reaction ; Receptors, Cytoplasmic and Nuclear ; genetics

Objective To investigate the methods and internal quality control ( IQC ) leucorrhea routine examinationin clinical laboratories of medical institutions in Guizhou Province.Methods In 2009, 97 clinical laboratories were randomly selected for the first investigation.At the same time, staffs in theinvestigated laboratories were educated on the importance of IQC.The second investigation of the same items was carried out in 2011 inthe same laboratories.The results of the two investigations were analyzed byChi-square test.Results 2009 and 2011 numbers of laboratories thoseonly used normal saline suspension method for leucorrhea examination were 17and 16 (χ2 =0.037, P >0.05 ) respectively, used bothnormal saline and 10%KOH suspension methodswere 16and 2(χ2 =12.003,P<0.01), used staining method were 64and 79(χ2 =5.488,P<0.05), both used suspension and staining methods were 60and 73(χ2 =4.041, P<0.05), used normal salinesuspension method combined with Wright stain and Gram staining methods were3and 28(χ2 =23.996,P<0.01) respectively.Numbers of Laboratoriespracticing IQC were 2and 88in 2009 and 2011 respectivly(χ2 =153.293,P <0.01).Conclusions Currently, the most common used method for leucorrhea routine examination is suspension.Through the investigations and education, the quality ofleucorrhea routine examination was improved in Guizhou Province.

Objective To establish genotyping methods for vitamin K epoxide reductase complex subunit 1 (VKORC1)and cytochrome P450 2C9(CYP2C9)based on pyrosequencing technique to detection of warfarin metabolizing enzyme related gene polymorphisms.Methods A total of 50 peripheral blood samples from healthy adults were collected and the whole blood genomic DNA was extracted.A set of biotin-labeled amplifi-cation primers and sequencing primers were designed respectively for three SNP sites:VKORC1 -1639 G>A,CYP2C9 430C> T and CYP2C9 1075A>C.After PCR amplification of the samples,pyrophosphoric acid se-quencing was conducted.And then the signal peaks form were combined to analyze and determine each sample genotype.Genotyping results were verified by Sanger sequencing,and the consistency of the two sequencing methods was compared.Results Genotypes of the three SNPs can be clearly determined according to the ba-ses and height of the signal peaks.Among the 50 samples,there were 41 AA and nine AG for VKORC1 -1639G>A,accounting for 82% and 12% respectively,and there were 45 *1/*1,five *1/*3 for CYP2C9, accounting for 90% and 10% respectively,no CYP2C9*2 allele detected.Genotype results detected by pyrose-quencing and Sanger sequencing were consistent with each other.Conclusion In SNP genotyping,Pyrose-quencing has the advantages of convenience,time-saving,cheap with accurate and reliable results,which can quickly determine the genotypes of CYP2C9 and VKORC1.

OBJECTIVETo investigate the spectrum of β -thalassemia mutations in Guizhou Province.

METHODSFor 542 individuals suspected to have β -thalassemia by decreased mean corpuscular volume (MCV) and corpuscle hemoglobin (MCH) by routine blood test and hemoglobin electrophoresis, reverse dot blot hybridization (RDB) was performed to detect 17 known β -thalassemia mutations, including 8 common and 9 rare mutations. For cases where no mutation was identified, the entire human β -globin gene was screened to find other rare mutations. The distribution and frequencies of detected β -thalassemia mutations were then analyzed.

RESULTSA total of 460 individuals were diagnosed as β -thalassemia by DNA analysis, which included 352 heterozygotes, 67 compound heterozygotes and 41 mutant homozygotes. A total of 12 β -thalassemia mutations were detected in these individuals. The mutations have ranked from high to low frequency as: CD17 (40.74%), CD41-42 (33.69%), IVS-II-654 (13.76%), -28 (3.70%), β E (3.35%), CD71-72(1.94%), CD43 (1.06%), IVS-I-1 (0.71%), CD27-28 (0.35%), -29(0.35%), CAP (0.18%), and CD121 (0.18%). The former six mutations have accounted for 97.18% of all. CD121 (GAA> TAA) detected from a heterozygote, as a dominant mutation, has been firstly found in the Chinese population.

CONCLUSIONThe spectrum of β -thalassemia in Guizhou Province showed certain distinct characteristics, with CD17 being the most common mutation. The newly discovered mutation of CD121 has expanded the spectrum of β -thalassemia in Chinese population. Our result may provide valuable information for the prevention and control of β -thalassemia in Guizhou.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; China ; DNA Mutational Analysis ; Female ; Humans ; Infant ; Leukosialin ; genetics ; Male ; Middle Aged ; Mutation ; Platelet Membrane Glycoprotein IIb ; genetics ; Receptors, Interleukin-1 Type I ; genetics ; Young Adult ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; ethnology ; genetics

OBJECTIVE: To improve the diagnosis and treatment of Kimura's disease (KD) by investigating its clinical characteristics, pathological features and complications. METHOD: The clinical data of 33 cases of KD were analyzed retrospectively. RESULT: Of 33 cases, 22 showed the mass on head and neck, while in the other cases, the mass distributed in the region of groin, axillary fossa, hilum of lung and mesentery. Regional lymph nodes were involved in 21 cases and major salivary glands were invaded in 8 cases. Twenty-three cases had typical peripheral eosinophilia, although only in 2 patients the quantity of serum total IgE increased markedly. Urine abnormalities happened to 7 cases, such as massive proteinuria (3 cases) and hematuria (2 cases). Among 6 cases which underwent bone marrow aspiration, 2 showed eosinophilia. Two cases were complicated with nephritic syndrome. Six cases were combined with local inflammation on head and neck and 2 cases were combined with malignant tumor. CONCLUSION Mass on the head and neck is the typical clinical manifestation in KD, with regional lymph nodes and major salivary glands involved most. Serum total IgE and histopathologic examination should always be done to confirm KD, especially in the cases with unknown eosinophilia increasing.
Adolescent ; Adult ; Aged ; Angiolymphoid Hyperplasia with Eosinophilia ; diagnosis ; pathology ; therapy ; Child ; Female ; Head ; pathology ; Humans ; Immunoglobulin E ; blood ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neck ; pathology ; Retrospective Studies ; Salivary Glands ; pathology ; Young Adult