OBJECTIVETo setup a quantitative assay for detection of methylation of SNCG CpG island in human tissue samples.

METHODSMethylation status of the 16 tested CpG sites within the CpG island was analyzed by bisulfite-clone-sequencing for 2 gastric carcinoma cell lines, 2 normal gastric mucosa samples, and 2 pairs of primary gastric carcinomas and their corresponding non-neoplastic tissues, respectively.

RESULTSThe methylation of -88 and other four CpG sites was well correlated with the methylation of the overall CpG island. Thus, a combined bisulfite-restriction assay (COBRA) was developed based on the enzyme AciI, which digested the only one GCGG sequence in the PCR products of the methylated CpG island, but not the GTGG in the demethylated one. The digested fragments (144 bp and 85 bp) and undigested fragment (229 bp) could be completely separated by denaturing high performance liquid chromatography (DHPLC). According to the peak areas of these fragments, the proportion of the methylated copies of the SNCG CpG island was calculated easily. The result of the COBRA-DHPLC assay was reproducible and consistent with that of clone-sequencing.

CONCLUSIONA COBRA-DHPLC assay is setup successfully for quantification of methylation of the SNCG CpG island.

Cell Line ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; CpG Islands ; DNA Methylation ; Humans

OBJECTIVE: To seek a exact method of estimating postmortem interval (PMI). METHODS: This study detected the concentration of zincum(Zn) and nickel(Ni) in vitreous humor of rabbit at hour 96 after death and explored the relationship between their concentration and PMI using a method ICP-MS. RESULTS: The concentration of Zn and Ni in vitreous humor of rabbit at hour 24 after death were related to PMI significantly; The formulae of the relationship between PMI and Zn concentrations is y = 0.1404x2 - 1.3351x + 3.8298 (within 24 h; R2 = 0.9202). The formula of the relationship between PMI and Ni concentrations is y = 0.0043x2 - 0.0596x + 0.2665(within 24 h; R2 = 0.9103). CONCLUSION The concentration of Zn and Ni in vitreous humor of rabbit may be a reference indicator to estimate early PMI.
Animals ; Female ; Forensic Medicine ; Male ; Nickel/analysis* ; Postmortem Changes ; Rabbits ; Time Factors ; Vitreous Body/chemistry* ; Zinc/analysis*

OBJECTIVETo investigate relationship between methylation status of the APC and Bikunin CpG islands and clinicopathological characteristics of breast carcinomas.

METHODSThe methylation status of APC and Bikunin CpG islands in 152 sporadic breast carcinoma samples were analyzed by methylation specific PCR.

RESULTS40.8% of breast carcinomas examined showed methylated signals for the APC. The methylation frequency of APC was significantly correlated to the tumor size (chi(2) = 4.041; P = 0.044), but not to patients' age, pathologic type of tumor, clinical stage, histological grade, lymph node metastasis and the status of estrogen or progestogen receptor. In addition, 24.6% of carcinoma samples examined revealed strong methylated signals for Bikunin. No significant correlation was found between the aberrant methylation of Bikunin and the clinicopathological characteristics of sporadic breast carcinomas.

CONCLUSIONThe aberrant methylations of APC and Bikunin are frequent events during breast carcinogenesis. APC methylation might play a role in the progression of breast cancer.

Breast Neoplasms ; genetics ; pathology ; CpG Islands ; DNA Methylation ; DNA, Neoplasm ; genetics ; Female ; Follow-Up Studies ; Humans

OBJECTIVE: To study the occurrence of serious adverse events (SAEs) related to chemotherapy with CCCG-ALL-2015 regimen in children with acute lymphoblastic leukemia (ALL) and the risk factors for death after the SAEs. METHODS: A retrospective analysis was performed on the medical data of 734 children with ALL. They were treated with CCCG-ALL-2015 regimen from January 2015 to June 2019. The occurrence of SAEs during the treatment was investigated. The children with SAEs were divided into a death group with 25 children and a survival group with 31 children. A multivariate logistic regression analysis was used to analyze the risk factors for death after the SAEs. RESULTS: Among the 734 children with ALL, 56 (7.6%) experienced SAEs (66 cases) after chemotherapy, among which 41 cases occurred in the stage of remission induction therapy. Of all 66 cases of SAEs, 46 (70%) were infection-related SAEs, including 25 cases of septic shock (38%), 20 cases of severe pneumonia (30%), and 1 case of severe chickenpox (2%), and 87% of the children with infection-related SAEs had neutrophil deficiency. The most common infection sites were blood and the lungs. The most common pathogens were Gram-negative bacteria, viruses, fungi, and Gram-positive bacteria. There were 16 cases (24%) of hemorrhage-related SAEs, with 11 cases of gastrointestinal bleeding (17%), 4 cases of pulmonary bleeding (6%), and 1 case of intracranial bleeding (2%). Of all 734 children with ALL, 66 (9.0%) died, among whom 25 died due to SAEs. The treatment-related mortality rate was 3.4%, and infection (72%) and bleeding (24%) were the main causes of death. Severe pneumonia was an independent risk factor for treatment-related death in ALL children (OR=4.087, 95%CI: 1.161-14.384, P=0.028). CONCLUSIONS SAEs often occur in the stage of remission induction therapy, and infection-related SAEs are more common in ALL children accepting chemotherapy with CCCG-ALL-2015 regimen. The development of severe pneumonia suggests an increased risk for death in these children.
Antineoplastic Agents ; adverse effects ; Child ; Gram-Negative Bacteria ; Humans ; Neutrophils ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Retrospective Studies ; Risk Factors

This study was aimed to investigate the relationship between endostatin and vascular cell adhesion molecule-1 (VCAM-1) expressions on bone marrow stromal cells (BMSC) in mice after bone marrow transplantation (BMT) and effect of ligustrazine on their expressions. The mice were randomly divided into 3 groups: normal group (without treatment), saline group (control of BMT) and ligustrazine group (BMT + ligustrazine). BMT mouse models were established. The normal group was not treated, the saline group was given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the ligustrazine group was given ligustrazine (0.2 ml/mouse, twice a day) also through gastric tube. On day 7, 14, 21 and 28 after BMT, mice were killed by euthanasia. The expression levels of endostatin and VCAM-1 in bone marrow stromal cells were detected by immunohistochemistry and RT-PCR analysis respectively. The results showed that the endostatin protein mainly expressed in nuclei of BMSCs, the VCAM-1 protein mainly expressed in plasma of BMSCs. On day 7, 14, 21 after BMT the expression levels of endostatin mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), while their expression levels in ligustrazine group were lower than that in saline group. On day 28 the expression levels in saline group returned to normal, while the expression levels in ligustrazine group not were normalized. On day 7, 14, 21 after BMT the expression levels of VCAM-1 mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), but their expression levels in ligustrazine group were significantly lighter than that in saline group (P < 0.01 or P < 0.05). On day 28 the VCAM-1 expression level in ligustrazine group returned to normal, while its expression level in saline group not were normalized. The difference between these two groups was significant (P < 0.01). Correlation analysis revealed that there was a negative correlation between endostatin and VCAM-1 expression in saline group, there was a positive correlation between endostatin and VCAM-1 expression in ligustrazine group. It is concluded that the endostatin can influence hematopoiesis in bone marrow by affecting VCAM-1 expression on BMSC and hindering connection between stromal cells and hematopoietic cells as well as extracellular stroma and hematopoietic cells, while ligustrazine can enhance the adhesion molecule expression on stromal cell surface of bone marrow in BMT-mice, accelerate the homing and proliferation of HSPC in bone marrow after BMT, meanwhile can promote the repair of bone marrow microenvironment, accelerate hematopoietic reconstitution of bone marrow after BMT through feedback regulation of endostatin expression of BMSC in BMT-mice.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Marrow Transplantation ; Endostatins ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Stromal Cells ; metabolism ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVE: To compare the predictive value of Ees/Ea in patients with pulmonary arterial hypertension based on right cardiac catheterization and cardiac magnetic resonance examination. METHODS: A total of 50 pulmonary arterial hypertension patients confirmed by right heart catheterization were retrospectively reviewed and followed up 2 years. The relationship between the baseline clinical data and survival prognosis of patients with pulmonary arterial hypertension were analyzed. The predictive value of baseline Ees/Ea for prognosis in patients with pulmonary arterial hypertension were compared. RESULTS: The Ees/Ea calculated by volume method was significantly correlated with the prognosis of patients with pulmonary arterial hypertension(OR =0.082, 95%CI 0.009-0.814, P=0.032). The two-year survival rate of group Ees/Ea>0.67 was significantly higher than that of Ees/Ea≤0.67(86.7% vs. 50%, P=0.003). Conclution Ees/Ea calculated by volume method is an independent influence factor of the survival prognosis of patients with pulmonary arterial hypertension. When Ees/Ea≤0.67, the patients with pulmonary arterial hypertension is more likely to deteriorate.

OBJECTIVE: To seek a exact method of estimating Postmortem interval (PMI). METHODS: This study was preformed to investigate the relationship between postmortem interval and absorbance in vitreous humor of rabbit after death. The absorbance in vitreous humor of 48 rabbits after death were investigated with Model 754 spectrophotometer in apt wavelength (420 nm). RESULTS: There exists positive linear regression association between postmortem interval (Y) and absorbance in vitreous humor (X) (r = 0.98327, P < 0.05), during rabbits after death 0 to 72 hours. The formula of linear regression is Y = 453.30 X + 0.75 (Y = postmortem interval = PMI, X = absorbance in vitreous humor). CONCLUSION The absorbance in vitreous humor can be as reference indicator to estimate PMI within hour 72.
Animals ; Female ; Male ; Postmortem Changes ; Rabbits ; Spectrophotometry ; Time Factors ; Vitreous Body

A GeXP based multiplex PCR assay was developed to simultaneously detect six different chicken respiratory viruses including H5, H7, H9 subtypes of avian influenza virus(AIV), new castle disease virus (NDV), infectious bronchitis virus(IBV) and infectious laryngotracheitis virus(ILTV). According to the conserved sequences of genes of each pathogen, seven pairs of specific primers were designed, and the reaction conditions were optimized. The specificity and accuracy of GeXP were examined using samples of single and mixed infections of virus. The sensitivity was evaluated by performing the assay on serial 10-fold dilutions of cloned plasmids. To further evaluate the reliability, thirty-four clinical samples were detected by GeXP. The corresponding specific fragments of genes were amplified. The detection limit of GeXP was 10(2) copies/microL when all of 7 pre-mixed plasmids containing target genes of six chicken respiratory viruses were present. In the detection of thirty-four clinical samples, the results of GeXP were accorded with the viral isolation completely. In conclusion, this GeXP assay is a rapid, specific, sensitive and high-throughput method for the detection of chicken respiratory virus infections. It can be applied in rapid differential diagnosis for clinical samples, and also provide an effective tool to prevent and control chicken respiratory diseases with similar clinical symptoms.
Animals ; Chickens ; Influenza A virus ; classification ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; diagnosis ; virology ; Multiplex Polymerase Chain Reaction ; methods ; Poultry Diseases ; diagnosis ; virology ; Respiratory Tract Infections ; diagnosis ; veterinary ; virology

OBJECTIVETo compare the diagnostic performance of prospective electrocardiogram (ECG)-triggered computed tomography coronary angiography (CTCA) versus retrospective ECG-gated CTCA.

METHODSPatients with suspected coronary artery disease divided into two groups which underwent 64-slice CTCA with prospective ECG-triggered or retrospective ECG-gated scanning (n = 100 each, HR < or = 65 bpm). Multi-planar reconstruction (MPR), curved-planar reconstruction (CPR), maximum intensity projection (MIP) and volume rendering (VR) were made to demonstrate the coronary arteries. The image quality was defined as excellent, good and poor by motion and stair-step artifacts. Individual radiation exposure dose was estimated from the dose-length product.

RESULTSThe mean effective radiation dose of prospective ECG-triggered CTCA [(2.81 +/- 0.48) mSv] was significantly lower than that of retrospective ECG-gated CTCA [(10.16 +/- 1.09) mSv, P < 0.01]. Segments of diagnostic image quality (95.2%, 1165/1224) and non-diagnostic coronary segments (4.8%, 59/1224) in prospective ECG-triggered group were similar as those of retrospective ECG-gated group (94.1%, 1186/1261 and 5.9%, 75/1261, all P > 0.05).

CONCLUSIONThough the radiation exposure dose required is significantly lower, the diagnostic performance of prospective ECG-triggered 64 slice CTCA is comparable with that of retrospective ECG-gated 64 slice CTCA on patients with stable heart rates up to 65 bpm.

Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; methods ; Electrocardiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Retrospective Studies ; Tomography, X-Ray Computed ; methods

OBJECTIVETo study the characteristics of amplitude integrated electroencephalography (aEEG) in preterm infants and changes of maturation with gestational age.

METHODSaEEG monitoring was done within 3 days of age with domestically produced digital aEEG set (CFM3000). Duration of each recording was at least 4 hours. The continuity, sleep-wake cycle, voltage and bandwidth of all aEEG tracing were analyzed.

RESULTSThe percent of continuity background increased from 30% of 28 weeks to 85.7% of 36 weeks (χ(2) = 28.2, P = 0.026); the percent of mature sleep-wake cycle increased from 10% of 28 weeks to 100% of 36 weeks (χ(2) = 192.4, P < 0.01). Low bound voltage increased with gestational age, from (6.8 ± 1.7) µV (28 w) to 9.7 - 10.1 µV (35 - 36 w) (F = 11.4, P < 0.01). Bandwidth of the narrow band decreases gradually with gestational age, from 1.45 cm (28 w) to (0.86 ± 0.24) cm (36 w) (F = 8.731, P < 0.01). The correlation coefficient for continuity, sleep-wake cycle, low bound voltage and bandwidth of narrow band, and total scores were 0.32, 0.81, 0.38, 0.55 and 0.78 respectively (P < 0.05).

CONCLUSIONThe older the gestational age of infants at birth, the more mature the aEEG pattern, manifested as increased continuity and sleep-wake cycle, the higher low bound voltage and more narrowed bandwidth with increased gestational age.

Age Factors ; Electroencephalography ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; physiology ; Male