Objective To investigate the changes of human epidermal stem cells after transfected with human telomerase reverse transcriptase(hTERT)gene.Methods The plasmid pIRES2-EGFP and plasmid pIRES2-EGFP-hTERT encoding hTERT were transfected into in vitro cultured human fetal epidermal stem cells by liposome-mediated transfection.Then,the positive cells were selected with G418.The mRNA and protein expressions of hTERT were detected by reserve transcriptase-polymerase chain reaction(RT-PCR)and Western blot.The telomerase activity and the proliferation and cycle of human epidermal stem cells were detected by telomeric repeat amplification protocol(TRAP)-ELISA and flow cytometry respectively.Results RT-PCR and Western blot techniques detected weak mRNA and protein expressions of hTERT gene in untransfected and vacant vector transfected cells but high level of mRNA and protein expressions of hTERT gene in pIRES2-EGFP-hTERT transfected cells.Compared with untransfected and vacant vector transfected cells,the pIRES2-EGFP-hTERT transfected cells had higher telomerase activity,with lower proportion of cells at G_0/G_1 phase,higher proportion of cells at S and G_2/M phases and enhanced proliferation ability.Conclusion Transfection with hTERT gene can markedly enhance mRNA and protein expressions,telomerase activity and proliferation ability of hTERT gene of human epidermal stem cells euhured in vitro.

A rapid method was developed for the determination of 5 common fatty acids, palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid in industrial oleic acid based on ultra_performance convergence chromatography_mass spectrometry ( UPC_MS) . The sample was dissolved by n_hexane, followed by clean_up of extract using 0. 22 μm organic phase filter. The fatty acids were separated in 3 min on the column of Acquity UPC2 BEH 2_EP by gradient elution with carbon dioxide and methanol/acetonitrile (1∶1, V/V) system, and finally detected by MS detector in ESI- mode. Through the optimization of UPC2_MS condition, the reasonable linearity was achieved for all the analytes over the range of 0. 5-100 mg/L with the correlation coefficients ( R2 ) greater than 0. 9985. The recoveries for five fatty acids at three spiked levels were in the range from 89 . 3% to 106 . 67% with relative standard deviations of 0 . 8%-3 . 0%. The limits of detection for target compounds in the method ranged from 0. 07 mg/L to 0. 26 mg/L. The real sample analysis showed that this method was simple,fast and had a good separation effect. There was no need of derivatization for fatty acid samples. This work would provide a fast and effective detection method for UPC2 technology in oil related research field.

Radiation-induced brain injury (RBI) is a common long-term complication of radiotherapy for nasopharyngeal carcinoma (NPC) and seriously affects the quality of life and overall survival of patients. In the era of intensity-modulated radiation therapy (IMRT), the long-term complications after radiotherapy, especially RBI, are becoming increasingly concerning because a number of treated patients with NPC obtain long-term survival. At present, the understanding of RBI is still being explored, and its pathogenesis and treatment methods are continuously updated. This article reviews the research progress of RBI in patients with NPC.

Objective: To evaluate the role of tuberculin skin test prified protein derivative (PPD) in defining the screening scope of close contacts of tuberculosis cases in disposal of tuberculosis outbreak in schools. Methods: In a senior middle school in August 2019, 1 553 students of the grade two were tested by PPD because of a school tuberculosis outbreak. PPD results were compared to grade one students without any association with this tuberculosis epidemic, who were also tested by PPD when beginning school. Potential association between PPD distribution characteristics and tuberculosis case distribution were analyzed. Results: Twenty nine grade two students were diagnozed as tuberculosis infection, seven of which were PPD positive, and with the same MIRU-VNTR genotype. In grade one, 0.1 % (1/796) student showed strong PPD positive, 34.3% (273/796) student showed positive. For grade two students, significant higher rate of strong PPD positive [5.9% (45/757)], and PPD positive [52.0% (394/757)] were observed ( χ 2=45.71, 49.90, P <0.01). Proportion of strong PPD positive in the first floor of the teaching building ( 19.4 %), where tuberculosis cases clustered, was significantly higher than that in other floors ( χ 2=89.81, P <0.01); Number of strong PPD positive students increased with TB cases in each floor of the teaching building ( r =0.99, P <0.01). Proportion of strong PPD positive and PPD positive in floors of the dormitory, where TB cases lived, was significantly higher than in other floors ( χ 2=49.4, 64.9 , P <0.01). Number of strong PPD positive and PPD positive students increased with TB cases in each floor of the teaching building ( r =0.84, 0.56, P <0.01). Conclusion Strong PPD positive rate well reflects tuberculosis infection of close contacts, and is recommended for defining the scope of screening.

OBJECTIVETo investigate the mitochondrial injury in enterovirus 71 (EV71)-infected Vero cells and explore the possible mechanism.

METHODSA clinical isolate of EV71 was inoculated to Vero cells and the EV71 antigen was detected by immunofluorescence assay. The morphological changes of Vero cells were observed using optical microscopy and transmission electron microscopy. The diameter and area density of the viral particles and the ratio and area density of vacuolated mitochondria in the cells were measured on the ultrastructural images.

RESULTSEV71-infected Vero cells underwent obvious changes and to a spherical morphology followed by cell death EV71 particles were detected in the cytoplasm by immunofluorescence. Ultrastructurally, the infected cells contained a large number of viral particles in the cytoplasm, with a clustered distribution and lattice-like arrangement. The diameter of the particles were 16.3 nm and the mean area density was 38.3%. Most of the mitochondria presented with swelling, vacuoles and degeneration. The ratio of the vacuolated mitochondria was 90.9% with a mean area density of 89.2%. Viral particles were also found in some mitochondria.

CONCLUSIONEV71 proliferates in the cytoplasm and invades the mitochondria of infected Vero cells leading to mitochondrial injury and cell death, suggesting that mitochondria are the targets for EV71 infection.

Animals ; Cercopithecus aethiops ; Cytoplasm ; virology ; Enterovirus ; Enterovirus Infections ; pathology ; Humans ; Mitochondria ; pathology ; virology ; Vero Cells ; virology

Objective A prospective cohort study was carried out to assess the mortality and potential risk factors for primary liver cancer (PLC) in Haimen city of Jiangsu province.Methods The cohort involved 89 789 adult residents aged 25-69 years.Upon the entry of this project,each subject was asked to complete a questionnaire and to provide a blood sample of 10 ml.Surface antigen of hepatitis B virus (HBsAg) was tested by radioimmunoassay.All the subjects were followed-up every year for vital statistics and death certificate information until 2014.Cox proportional hazard model was used to estimate the hazard ratio (HR) and 95％ confidence interval (95％ CI) for PLC mortality associated with HBsAg status and other risk factors.Results During the 1 299 611 person-years of follow-up,a total of 2 583 PLC cases were identified,including 2 149 men and 434 women.Mortality of the PLC for men and women were 247.80/100 000 person-years and 100.38/100000 person-years,respectively.Among those who died of PLC,73.87％ had been tested HBsAg positive.HBV infection seemed the predominant risk factor for PLC and the HRs were 15.97 for men (95％CI:14.29-17.85) and 21.63 for women (95％CI:16.16-28.96) respectively.Ageing,cigarette smoking,previous history of hepatitis,and family history of HCC were factors associated with the increased risk for PLC.Conclusion People living in Haimen city had a high risk on PLC.HBV infection appeared the most important risk factor for HCC mortality in this area.

Objective: To investigate the diagnosis and treatment of synchronous double primary malignant tumours of the tongue and lung. Methods: A case of adenoid cystic carcinoma（ACC） and lung adenocarcinoma with double primary malignancy was retrospectively analyzed. Results: The tumor of patient′s tongue base gradually grew. MRI showed multiple enlarged lymph nodes on both sides of neck. CT of the chest showed obvious lesions in the anterior basal segment of the right lower lobe. The pathological biopsy of the tongue mass identified ACC, and pathological biopsy of the lung mass identified lung adenocarcinoma. The tongue and lung tumors were both surgically resected, and the tongue defect was repaired at the same time. No residue was found after surgery, and no recurrence was found during the follow-up period. The aesthetic and functional restoration of the lingual region was good. Conclusion There are few cases of adenoid cystic carcinoma and lung adenocarcinoma with double primary malignancies, and the related diagnosis and treatment are very difficult; the simultaneous removal of double primary malignant tumors may achieve good prognosis.