With the development of sequencing technology, the cost of whole-genome sequencing was significantly declined.Meanwhile, with the application of combined whole-genome sequencing with epigenetic analysis on methylation and histone acetylation, the comprehensive and systematic analysis of numerous samples became a reality and we are able to re-understand the genesis and development of cancer. New ideas are emerging in comparative genomics research methods, from comparison of genomes among different individuals to horizontal self-comparison of different tissues and vertical self-comparison of genomes recently.Individualized diagnosis and treatment of cancer has shown a bright future.
Genome, Human ; genetics ; Humans ; Neoplasms ; genetics ; therapy ; Precision Medicine ; Sequence Analysis, DNA ; methods

OBJECTIVETo develop a solid phase PCR method by covalent single point immobilization for recycle utilization of human genome.

METHODSPolymethacrylamide gel was selected as a solid PCR carrier based on DNA-hydrogel copolymer chemistry presented by Mirzabekov. (CH2)6NH2 amino-modified PCR product and randomly fractured formic acid-modified plasmid pGEM-T-HLA-G were used as templates. The specificity of the attachment chemistry was characterized by acrylamide gel electrophoresis, and the thermal stability of method was demonstrated by PCR. This method was applied for the recycle utilization of human genome. Sequencing was used to exclude the possibility of introduced mutations during modification and immobilization procedures.

RESULTSThe PCR detections of plasmid DNA and human genome DNA immobilized by polymethacrylamide gel was successful. The thermal stability of method was successfully demonstrated by performing PCR after 16 rounds of standard 36 PCR cycles. And the sequencing was found no mutation.

CONCLUSIONThe DNA immobilization method with polymethacrylamide gel as a solid phase carrier is stable and specific, which can be a possible approach for realizing recycle utilization of human genome for whole-genome sequencing and SNP detection.

Electrophoresis, Polyacrylamide Gel ; Genome, Human ; Humans ; Hydrogels ; Immobilized Nucleic Acids ; analysis ; Polymerase Chain Reaction ; methods

A 33 years old male patient who suffered from a flame burn of 88% total body surface area was admitted to our hospital on November 28th, 2016. During his hospitalization, we repeatedly performed central vein catheterization in internal jugular veins, subclavian veins, or femoral veins for fluid transfusion. We incidentally found bilateral internal jugular vein thrombosis by performing a point-of-care ultrasound examination before catheterizing sometime. We treated the patient by avoiding catheterization in the affected internal jugular veins, anticoagulating with low molecular weight heparin, closing the wounds with skin autografting, and guiding the patient to practice functional exercise. The thrombus disappeared in the end. The patient was cured and discharged 3 months post burn.

OBJECTIVE: To test the expression of NF-kappaB/p65, MMP-3 and MMP-9 in nasopharyngeal carcinoma (NPC), and analyze their relationship and the clinical significance. METHOD: All samples were examined for the expression of NF-kappaB/p65, MMP-3 and MMP-9 by streptavidin-peroxidase immunohistochemistry. Chi2-test and Spearman-test were used to exam the correlation within them and the expression difference between nasopharyngeal carcinoma and normal nasopharyngeal mucosa. RESULT: Positive expression rate of NF-kappaB/p65 in nasopharyngeal carcinoma was 78.0%; positive expressions rate of MMP-3 and MMP-9 in nasopharyngeal carcinoma were 75.6%. There was positive relationship between NF-kappaB/p65, MMP-3, MMP-9 and N stage. The expression of NF-kappaB/p65 showed a significant positive correlation with the expression of MMP-3 and MMP-9. MMP-3 also had a significant positive correlation with MMP-9. CONCLUSION The result suggests that NF-kappaB/p65, MMP-3 and MMP-9 are highly expressed in nasopharyngeal carcinoma cells. Their expression is correlated with lymph nodes metastasis, but do not have relationship with sex, age, pathological classification and clinical stage. Each of them has positive correlation with the others.
Adult ; Aged ; Female ; Humans ; Male ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; NF-kappa B ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Transcription Factor RelA ; metabolism

OBJECTIVETo investigate the associations between epidermal growth factor receptor (EGFR) gene mutations and serum tumor markers in advanced lung adenocarcinomas.

METHODSWe investigated the association between EGFR gene mutations and clinical features, including serum tumor marker levels, in 97 advanced lung adenocarcinomas patients who did not undergo the treatment of EGFR tyrosine kinase inhibitors. EGFR gene mutation was detected by real-time PCR at exons 18, 19, 20, and 21. Serum tumor marker concentrations were analyzed by chemiluminescence assay kit at the same time.

RESULTSEGFR gene mutations were detected in 42 (43%) advanced lung adenocarcinoma patients. Gender (P=0.003), smoking status (P=0.001), and abnormal serum status of carcinoembryonic antigen (CEA, P=0.028) were significantly associated with EGFR gene mutation incidence. Multivariate analysis showed the abnormal CEA level in serum was independently associated with the incidence of EGFR gene mutation (P=0.046) with an odds ratio of 2.613 (95% CI: 1.018-6.710). However, receiver operating characteristic (ROC) curve analysis revealed CEA was not an ideal predictive marker for EGFR gene mutation status in advanced lung adenocarcinoma (the area under the ROC curve was 0.608, P=0.069).

CONCLUSIONSEGFR gene mutation status is significantly associated with serum CEA status in advanced lung adenocarcinmoas. However, serum CEA is not an ideal predictor for EGFR mutation.

Adenocarcinoma ; blood ; genetics ; Biomarkers, Tumor ; blood ; Female ; Humans ; Lung Neoplasms ; blood ; genetics ; Male ; Middle Aged ; Mutation ; ROC Curve ; Real-Time Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; Retrospective Studies

OBJECTIVE: To explore the expression of CD23, CD19 on peripheral blood lymphocytes as well as its association with serum total IgE levels and nasal allergic symptoms in patients with allergic rhinitis (AR). METHOD: Symptom scores were evaluated in 46 AR patients, expression of CD23, CD19 on peripheral blood lymphocytes were measured by flow cytometry, and serum total IgE levels were determined by immune chemiluminescence. Thirty two healthy individuals were enrolled as controls. RESULT: The percentage of CD23+, CD19+ and CD23+/ CD19+,on peripheral blood lymphocytes in AR patients were 11.6 +/- 1. , 22.8 +/- 3.3,10.2 +/- 1.7, respectively. Higher frequencies of CD23+, CD19+, and CD23+/CD19+ were found in AR patients compared with controls (P < 0.05). There were positive correlations between expression rates of CD23+, CD19+, CD23+/CD19+ and levels of serum total IgE, nasal allergic symptom scores, respectively. CD23+/CD19+ demonstrated greater correlations with serum total IgE and nasal allergic symptom (r = 0.65 and 0.49, P < 0.05) than CD23+ and CD19+ did. Correlation between CD23+/CD19+ and nasal allergic symptom scores was greater than the corresponding correlations of serum total IgE (r = 0.33, P < 0.05). CONCLUSION CD23 and CD19 are important factors that associated with serum total IgE in the pathogenesis of AR, Analysis on the expression of CD23+/CD19+ on peripheral blood lymphocytes is helpful for evaluating the severity of AR.
Adolescent ; Adult ; Antigens, CD19 ; blood ; Case-Control Studies ; Female ; Humans ; Immunoglobulin E ; blood ; Lymphocyte Count ; Lymphocytes ; metabolism ; Middle Aged ; Receptors, IgE ; blood ; Rhinitis, Allergic, Perennial ; blood ; Young Adult

OBJECTIVE: To study the expression of epidermal growth factor receptor (EGFR), C-erbB-2 and its relationship with cell proliferation in nasopharyngeal carcinoma. METHOD: Expression of C-erbB-2, EGFR and proliferating cell nuclear antigen (PCNA) were detected with immunohistochemical staining in 32 nasopharyngeal carcinoma samples and 12 chronic inflammatory nasopharyngeal tissue samples. RESULT: The positive rate of EGFR,C-erbB-2, and PCNA expression in nasopharyngeal carcinoma was 65.6%, 37.5%, and (42.5 +/- 22.6)%, respectively, which was significantly higher than that in chronic inflammatory nasopharyngeal tissue (P < 0.05). There were positive correlations between the positive rate of EGFR, C-erbB-2, and PCNA expression and histopathological stage. The co-expression of C-erbB2 and EGFR was found in 62.5% (20/32) nasopharyngeal carcinoma samples. There was a positive correlation between C-erbB-2 and EGFR expression (r = 0.38, P < 0.05). The highest percentage of PCNA expression was found in carcinoma samples with co-expression of C-erbB and EGFR. CONCLUSION C-erbB-2, EGFR might have synergetic effect in the development and progress of nasopharyngeal carcinoma. The co-expression of C-erbB-2 and EGFR closely correlates with cell proliferation status.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Proliferation ; ErbB Receptors ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; metabolism ; Receptor, ErbB-2 ; metabolism ; Respiratory Mucosa ; metabolism ; Young Adult

OBJECTIVETo explore the origin and mechanism of small supernumerary marker chromosomes (sSMC) in order to facilitate genetic counseling.

METHODSChromosome karyotypes of two fetuses and their immediate family members were analyzed by conventional G banding. High-throughput whole genome sequencing was used to determine the origin of sSMCs.

RESULTSFetus 1 was shown to have a karyotype of 47,XY,+mar but with normal FISH and B ultrasound findings. Its father also had a 47,XY,+mar karyotype with normal FISH results and clinical phenotype. High-throughput genome sequencing revealed that fetus 1 and its father were both 46,XY,dup(21)(q11.2;q21.1) with a 6.2 Mb duplication of the long arm of chromosome 21. The fetus was born with normal phenotype and developed well. Its grandmother also had a karyotype of 46,XX,t(15;21)(q13;p13) with normal FISH result and clinical phenotype. The karyotypes of its mother and grandfather were both normal. Analysis of fetus 2 showed a 47,XY,+mar karyotype with normal FISH results. High-throughput genome sequencing suggested a molecular karyotype of 46,XX. The fetus was born with normal phenotype and developed well. The karyotypes of its parents were both normal.

CONCLUSIONConsidering their variable origins, identification of sSMC should combine conventional G banding analyses with high-throughput whole genome sequencing for precise delineation of the chromosomes.

Adult ; Amniotic Fluid ; chemistry ; Chromosome Banding ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Cytogenetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Markers ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis ; Young Adult

OBJECTIVETo analyze the deletion region for two fetal cases with large Yq deletions in order to provide genetic counseling and prenatal diagnosis.

METHODSFor both cases, amniotic fluid samples were cultured and analyzed with G banding and fluorescence in situ hybridization (FISH). Multiplex polymerase chain reaction was also carried out to amplify 15 sequence tagged sites (STS) of azoospermia factor (AZF) on the Y chromosome.

RESULTSFor both samples, the karyotypes were determined as 46,X,del(Y)(pter→q11:). No heterochromatin was found in C band. The karyotypes of their fathers were 46,XY, and heterochromatin was found in C band. STS analyses suggested that only sY82, sY84 and sY86 in AZFa were amplifiable while the other 12 STS were negative in amniotic fluid for the first case, which indicated deletions of AZFb, AZFd and AZFc. No AZF deletion was found in its father. For the second case, all 15 STS were amplifiable in the amniotic fluid, suggesting no AZF deletion. No AZF deletion was found in its father too.

CONCLUSIONConventional karyotyping combined with FISH and molecular genetics techniques can enable characterization of AZF microdeletions and facilitate genetic counseling and prenatal diagnosis.

Adult ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Counseling ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Pregnancy ; Prenatal Diagnosis

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Nails ; injuries ; surgery ; Suture Techniques