This study aimed to construct recombinant adenovirus expressing Epstein-Barr nuclear antigen 1 (EBNA1) against nasopharyngeal carcinoma (NPC). The C-terminal region fragment of the ebna1 gene of Epstein-Barr virus was amplified from the standard strain B95-8 by polymerase chain reaction (PCR). The gene fragment was inserted into the pDC316 shuttle plasmid using the EcoRI and BgIII restriction enzyme sites. The pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells after sequencing. The soluble protein was extracted from HEK293 cells, which caused apparent cytopathic effects. The transcription and expression of the ebna1 gene were confirmed using flow cytometry and Western blotting. rAd-ebna1 titers were measured by the TCID50. rAd-ebna1 was injected into BALB/c mice at a dose of 2 x 10(8) VP per mouse, EBNA1 epitope-specific responses were measured at 1st, 2nd, 4th and 8th weeks post-immunization. The target fragment of ebna1 (939 bp) was obtained by PCR, and was in consensus with the sequence from the standard strain B95-8. Cytopathic effects were observed after the pDC316-ebna1 shuttle plasmid and pBHG helper plasmid were cotransfected into HEK293 cells. rAd-ebna1 was successfully recombined in HEK293 cells. EBNA1 protein was detected in HEK293 cells, rAd-ebna1 titers reached 10(8) TCID50/mL. Specific responses to CD4+ epitopes of EBNA1 were detected in the immunized mice. In conclusion, rAd-ebna1 was successfully constructed and induced specific responses to CD4+ epitopes of EBNA1 in immunized mice.
Adenoviridae ; genetics ; immunology ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; virology ; Epstein-Barr Virus Infections ; immunology ; prevention & control ; virology ; Epstein-Barr Virus Nuclear Antigens ; administration & dosage ; genetics ; immunology ; Genetic Vectors ; genetics ; immunology ; Herpesvirus 4, Human ; genetics ; immunology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Viral Proteins ; administration & dosage ; genetics ; immunology

BACKGROUNDThe lung functional status could be displayed on lung perfusion images. With the images, the radiotherapy plans of lung cancer could be guided to more optimized. This study aimed to assess quantitatively the impact of incorporating functional lung imaging into 3-dimensional conformal radiotherapy (3DCRT) and intensity-modulated radiation therapy (IMRT) planning for non-small cell lung cancer (NSCLC).

METHODSTen patients with NSCLC who had undergone radiotherapy were included in this study. Before radiotherapy, each patient underwent CT simulation and lung perfusion imaging with single photon emission computed tomography (SPECT). The SPECT images were registered with simulation planning CT and used to contour functional lung (lung-F) and non-functional lung (lung-NF). Two 3DCRT plans and two IMRT plans were designed and compared in each patient: two anatomic plans using simulation CT alone and two functional plans using SPECT-CT in addition to the simulation CT. Dosimetric parameters of the four types of plans were compared in terms of tumor coverage and avoidance of normal tissues. Total radiation dose was set at 66 Gy (2 Gy x 33 fractions).

RESULTSIn incorporating perfusion information in 3DCRT and IMRT planning, the reductions on average in the mean doses to the functional lung in the functional plan were 168 cGy and 89 cGy, respectively, compared with those in the anatomic plans. The median reductions in the percentage of volume irradiated with > 5 Gy, > 10 Gy, > 20 Gy, > 30 Gy and > 40 Gy for functional lung in the functional plans were 6.50%, 10.21%, 14.02%, 22.30% and 23.46% in 3DCRT planning, respectively, and 3.05%, 15.52%, 14.16%, 4.87%, and 3.33% in IMRT planning, respectively. No greater degree of sparing of the functional lung was achieved in functional IMRT than in 3DCRT.

CONCLUSIONFunction-guided 3DCRT and IMRT plannings both appear to be effective in preserving functional lung in NSCLC patients.

Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; pathology ; radiotherapy ; Humans ; Radiography ; Radiotherapy, Conformal ; methods ; Radiotherapy, Intensity-Modulated ; methods ; Tomography, Emission-Computed, Single-Photon ; methods

Objective The aim of our study was to investigate the influence of hexavalent chromium exposure on mRNA expression of cell cycle related genes in electroplating workers, and to provide population data for investigating the toxic mechanisms of hexavalent chromium. Methods A total of 155 cases of workers occupationally exposed to hexavalent chromium were selected, including 89 males and 66 females, and the average age of workers was 39.65±8.856 years old. Questionnaire was used to collect essential information of workers. Peripheral blood was collected from electroplating workers. The inductively couple plasma mass spectrometry (ICP-MAS) was used to measure total blood chromium content. The workers were divided into four groups according to the blood chromium content. After extracting total RNA from whole blood and reverse transcription, the mRNA expression levels of p16 and CDK6 genes were detected by real-time quantitative PCR. Meanwhile, the levels of blood chromium (BCr) and the mRNA expression of p16 and CDK6 genes were compared among four groups. The impact of BCr, smoking habits, drinking habits, gender on the mRNA expression of CDK6 gene was analyzed. Results The levels of BCr in group 1 to 4 were 0.04ppb, 0.47±0.29 ppb, 2.76±1.16 ppb, 9.36 ±4.38 ppb, respectively, and the difference between every two groups was significant (P<0.05) . The median of p16 gene expression in four groups was 4.22, 7.19, 7.47, and 14.60, respectively, and the difference between every two groups was not significant (P>0.05) . The mRNA expression levels of CDK6 gene in groups 2 to 4 were 15.05, 8.03 and 24.81, respectively, which were significantly higher than that in group 1 (P<0.05) . The results of logistic regression showed that the level of Bcr was the main influence factor, while smoking habits, drinking habits and gender had no obvious impact on the mRNA expression of CDK6 gene. Conclusions Long-term exposure of hexavalent chromium led to higher mRNA expression of CDK6 gene, and it may serve as a biomarker for workers occupationally exposed to hexavalent chromium.

The aim of this study was to investigate the expression of anaplastic lymphoma kinase (ALK) protein resulted from chromosome translocation in anaplastic large cell lymphoma (ALCL) and its relationship with the age and prognosis of patients with ALCL. The tissue microarray including 30 cases of ALCL and 2 normal control tissues were established, the expression of anaplastic lymphoma kinase (ALK) protein was detected by immunohistochemistry, the statistical analysis of detected results was carried out by SPSS software. The results showed that the ALK protein was expressed negatively in 2 cases of primary skin ALCL, but in 20 out of 28 cases of systematic ALCL the ALK protein was expressed positively and mainly located in cytoplasm and/or nucleus (71.4%). Clinically, the patients with ALK expression were younger than those without ALK expression (p < 0.05). The prognosis of patients with ALK expression was better than those without ALK expression (p < 0.05). It is concluded that there is a high incidence of ALK expression in ALCL, especially in younger group. ALK expression may be an useful and independent marker for the differential diagnosis and prognosis evaluation of ALCL.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 2 ; Female ; Humans ; Lymphoma, Large-Cell, Anaplastic ; enzymology ; genetics ; Male ; Middle Aged ; Prognosis ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Receptor Protein-Tyrosine Kinases ; Translocation, Genetic ; Young Adult

To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.
Animals ; Brain Ischemia ; drug therapy ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate the role of t (14; 18) chromosomal translocation and bcl-2 amplification in classification, clinical staging and prognostic evaluation of diffuse large B cell lymphoma (DLBCL).

METHODSSixty cases of DLBCL were included in this investigation. Microdissection of the lymphoma tissue was performed. Tissue microarray and in-situ fluorescence hybridization technique were used to detect t (14; 18) and bcl-2 amplification. The phenotypes of either germinal center B-cell-like (GCB) or non-germinal center B-cell-like (non-GCB) were determined by immunohistochemistry including CD20, CD10, bcl-6 and MUM1 (S-P method) using the tissue microarray format. Clinical staging and therapeutic response were obtained by medical record review. The relationships among different parameters were analyzed by appropriate statistical methods.

RESULTSAmong 60 cases of DLBCL, bcl-2/IgH was positive in 10 cases and bcl-2 gene amplification was detected in 18 cases. Overall, 29 (48.3%) cases were GCB and 31 (51.7%) cases were non-GCB. The t (14; 18) was seen in 8 (80.0%) cases of GCB and 2 (20.0%) of non-GCB. The difference was statistical significance (P = 0.031). Over-expression of bcl-2 was seen in all cases having both t (14; 18) and bcl-2 gene amplification. Of thirty-six patients who underwent routine CHOP treatment, bcl-2 gene amplification was seen in 13 cases. In these cases, the rates of complete remission, partial remission and no change were 3 (23.1%), 4 (30.8%) and 6 (46.2%) respectively, and the clinical stages were stage I - II (1 case, 7.7%) and stage III - IV (12 cases, 92.3%). The clinical stages and therapeutic response were significantly different between the bcl-2 amplification cases and those without (P = 0.046 and P = 0.019, respectively).

CONCLUSIONST (14; 18) and/or bcl-2 gene amplification can lead to an over-expression of bcl-2 protein. The bcl-2 gene amplification correlates with worse therapeutic efficacies and advanced clinical stages. The reason for the correlation between bcl-2 over-expression and the prognosis is unclear, although it may be explained by different mechanisms that lead to bcl-2 over-expression. Detection of t (14; 18) chromosome translocation by FISH can be helpful in the classification of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Female ; Gene Amplification ; Genes, bcl-2 ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; classification ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tissue Array Analysis ; Translocation, Genetic ; Young Adult

BACKGROUND: The application of coronary stents, especially drug-eluting stents (DESs), has made percutaneous coronary intervention (PCI) one of important therapeutic methods for CHD. DES has reduced the in-stent restenosis to 5％–9％ and signifi cantly improved the long-term prognosis of patients with CHD. The study aimed to investigate the long-term efficacy and safety of domestic drug-eluting stents (DESs) in patients with acute coronary syndrome (ACS). METHODS: All patients with ACS who had undergone successful percutaneous coronary intervention (PCI) in the First Affiliated Hospital of Zhengzhou University from July 2009 to December 2010 were included in this study. Patients were excluded from the study if they were implanted with bare metal stents or different stents (domestic and imported DESs) simultaneously. The included patients were divided into two groups according to different stents implanted: domestic DESs and imported DESs. RESULTS: In the 1683 patients of this study, 1558 (92.6％) patients were folowed up successfuly for an average of (29.1±5.9) months. 130 (8.3％) patients had major adverse cardiovascular events (MACEs), including cardiac death in 32 (2.1％) patients, recurrent myocardial infarction in 16 (1％), and revascularization in 94 (6％). The rates of cardiac death, recurrent myocardial infarction, revascularization, in-stent restenosis, stent thrombosis and other MACEs were not significantly different between the two groups (allP>0.05). Multivarite logistic regression revealed that diabetes mellitus (OR=1.75, 95％CI: 1.09–2.82,P=0.021), vascular numbers of PCI (OR=2.16, 95％CI: 1.22–3.83, P=0.09) and PCI with left main lesion (OR=9.47, 95％CI: 2.96–30.26,P=0.01) were independent prognostic factors of MACEs. The Kaplan-Meier method revealed that there was no significant difference in cumulative survival rates and survival rates free from clinical events between the two groups (allP>0.05). CONCLUSIONS: The incidences of clinical events and cumulative survival rates are not statistically different between domestic DESs and imported DESs. Domestic DES is effective and safe in the treatment of patients with ACS.

[Objective] To investigate the HIV infection status and behavioral characteristics of men who have sex with men (MSM) over the age of 50 in Taizhou during 2010-2017 HIV sentinel surveillance, analyze the related factors of HIV infection, and provide basis for developing targeted intervention strategies. [Methods] HIV sentinel surveillance data from 2010 to 2017 in Taizhou was used to analyze the demographic characteristics, HIV awareness, and sexual behaviors of MSM over the age of 50. Also, HIV infection status and related factors were analyzed by multivariate regression analysis. [Results]A total of 334 MSM aged over 50 were surveyed by HIV sentinel surveillance in 2010-2017, and the HIV positive rate was 14.1％. The average age of the 334 MSM was (59.6 ± 7.2) years, and those with junior high school education and below accounted for 76.0％, with awareness of AIDS knowledge being 85.6％. And78.7％ of them had anal sex with men in the past 6 months. Among them, 30.4％ used condoms each time. And 4.8％ of them had commercial sex with men and 37.1％ had sex with women in the past 6months. Multivariate analysis showed that syphilis-positiveness was a risk factor for HIV infection and awareness of HIV-related knowledge proved to be a protective factor for HIV infection. [Conclusion] The MSM over 50 years old in Taizhou has a high rate of HIV-positiveness and high-risk behaviors exist. It is imperative to strengthen dynamic HIV surveillance for this population and behavioral intervention to reduce HIV related risky behavior.

OBJECTIVETo investigate the effects of artificial liver support system(plasma exchange combined with continuous veno - venous hemodiafiltration, PE + CVVHDF) on Gc globulin in patients with liver failure.

METHODS81 patients with liver failure were divided into 4 groups according to the treatment protocols and indicators such as liver function and clinical symptoms. Totally 29 effective cases and 14 ineffective cases in the ALSS group versus 15 effective cases and 23 ineffective cases in the medical group were included. Finally the changes of Gc globulin were observed in four subgroups before and after treatment. The correlation between Gc globulin and IL-10, IL-4, IL-18, TNFa, endotoxin, NO, sVCAM-1and sICAM-1were analyzed by Pearson correlation analysis.

RESULTSThe effectiveness rate was 67.44% in ALSS group and 34.21% in the medical treatment (P less than 0.01). Gc globulin, one of liver cell protection proteins was notably increased following the artificial liver treatment as compared with the increase in the medical treatment (P less than 0.01). The time-response curve of Gc globulin level had a significant upward trend in the effective group as compared to no significant rise in the ineffective group. Moreover, the Gc globulin was negatively correlated with IL-4, IL-18, TNFa, SVCAM-1, SICAM-1 and NO. In contrast, no correlation existed between Gc globulin and IL-10. The treatment with artificial liver can improve the outcome of the patients with liver failure. The level of Gc globulin was correlated with the curative effect and thus may be used as a potential indicator for curative effect forcast in the patients with liver failure.

Aged ; Cell Adhesion Molecules ; blood ; Cytokines ; blood ; Female ; Humans ; Liver Failure ; blood ; surgery ; therapy ; Liver, Artificial ; Male ; Nitric Oxide ; blood ; Treatment Outcome ; Vitamin D-Binding Protein ; blood ; metabolism

Conservative treatment with high doses of progestin is an alternative to standard hysterectomy for young patients with early-stage endometrial adenocarcinoma who desire to preserve their fertility. Here we report a patient with well-differentiated early-stage endometrial adenocarcinoma and poor fertility potential who failed to become pregnant in two in vitro fertilization-embryo transfer cycles and suffered a relapse after conservative treatment. This case illustrates that assessment of fertility potential is critical at the time of initial evaluation and conservative treatment planning for patients with endometrial adenocarcinoma.
Adenocarcinoma ; drug therapy ; metabolism ; Adult ; Antineoplastic Agents, Hormonal ; Endometrial Neoplasms ; drug therapy ; metabolism ; Female ; Follicle Stimulating Hormone ; therapeutic use ; Gonadotropins ; therapeutic use ; Humans ; Infertility ; Pregnancy ; Progesterone ; therapeutic use ; Reproductive Techniques, Assisted