Objective To find an ideal method of DNA isolation from blood and especially from clotted blood and to minimize the volume of blood collected for laboratory and clinical tests.Methods DNAs were isolated from antiagglutinated and agglutinated blood samples from auricular veins of 30 healthy subjects. The DNAs of these samples were obtained by a nonenzymatic, nontoxic procedure optimized by us and determinated by agarose gel electrophoesis and PCR. Results The yields of DNA isolated from clotted blood and antiagglutinated blood were (40.2?8.86)mg DNA/L and (39.1?10.2)mg DNA/L, and purities were 1.87?0.11 and 1.92? 0.12. The DNAs that we isolated from all samples had high molecular weight and by PCR the dimorphism of ALU alleles of the 8th intron of t-PA was easy to be obtained, so they were complete and reliable. Conclusion This method is rapid, easy, efficient and nontoxic for isolation of DNA from clotted and fresh blood and meets requirements for clinical testing and molecular biology study.

OBJECTIVETo develop a novel thermal treatment modality against metastatic tumor, and to verify the hypothesis that the extent of tumor angiogenesis damage and tumor cell necrosis, accompanied with immune suppression cells relief is deterministic to enhance therapeutic effect in the thermal treatment.

METHODSThe thermal treatment system was developed in our laboratory. The treatment including hyperthermia and alternate treatment, were locally applied to 4T1 murine mammary carcinoma. The extent of tumor necrosis was examined. Further investigations were performed to study the changes of MDSCs in peripheral blood and spleen.

RESULTSThe alternate treatment caused more damage to tumor microvasculature and tumor cell necrosis. Immunosuppression cells significantly reduced in peripheral blood and spleen. Moreover, it highly increased the survival rate of tumor-bearing mice.

CONCLUSIONSThe greatest destruction of primary tumor induced by the alternate treatment led to a relief of immune suppression in tumor bearing mice, and significantly increased therapeutic effect, especially for metastatic tumor.

Animals ; Female ; Hyperthermia, Induced ; methods ; Mammary Neoplasms, Experimental ; immunology ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Myeloid Cells ; immunology ; Neoplasm Metastasis

Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs.
Animals ; Antibodies, Viral ; blood ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Proliferation ; Classical swine fever virus ; genetics ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Haemophilus parasuis ; genetics ; Immunization ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics

Objective: To investigate the value of MRI in the early diagnosis of diagnosis of dysbaric osteonecrosis. Methods: Labor hygiene investigation and occupation health were examined on 52 high pressure operating personnel, were selected for the examination of both shoulders, hips and knees with X-ray and CT scan. Results: The cystic sign in dysbaric osteonecrosis as an important imaging feature, which perform in the MRI examination for T1W I sequence showed low or slightly low signal and T2W I sequence showed high signal, and X-ray and CT have a lower detection rate than MRI. The Kappa consistency test showed a high consistency with the two methods. At the same time MRI examination also can discover the bone marrow cavity necrosis early pathological change. Conclusion MRI is an effective method for the diagnosis of early dysbaric osteonecrosis, which can improve the early diagnosis rate of dysbaric osteonecrosis.

Objective:To explore the clinical efficacy of CT-guided 125I radioactive seed implantation in the treatment of bone metastases. Methods:The data of 43 patients with bone metastases treated by seed implantation from June 2016 to July 2018 in the 3201 Hospital was retrospectively analyzed. The 52 lesions of 43 patients with bone metastases were treated with 125I radioactive seed implantation. The patients were followed up, and the efficacy was evaluated based on pain relief, quality of life improvement, and whole body bone imaging. Results:The number of pain relief patients in 43 patients with bone metastases at 1, 3, 7, 30, 60, 90, and 180 d after 125I radioactive seed implantation were 19, 32, 39, 36, 36, 34, and 31 cases; the pain relief rates were 44.19%, 74.28%, 90.70%, 83.72%, 79.07%, and 72.09%, respectively. The pain relief rate was highest at 7 d after seed implantation, and the pain degree after seed implantation was significantly lower than that before seed implantation, the difference was statistically significant (Z = -5.216, P < 0.05). The difference in quality of life between patients before and after 125I radioactive seed implantation was statistically significant (Z = -4.308, P < 0.05). The re-examination in 3 to 6 months after treatment showed that the metabolism of 45 bone metastases lesions was reduced and shrunk, and the efficiency rate was 86.54% (45/52). Conclusion:125I radioactive seed implantation has good curative effect in the treatment of patients with bone metastases, which can effectively relieve pain, improve local control rate, and improve patients' quality of life.

Objective:To investigate the expression profile change of long non-coding RNA (IncRNA) and mRNA in plasma samples before and after drug resistance of gefitinib for non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR)-sensitive gene mutation treated, and to screen out RNA molecule related to gefitinib-resistance.Methods:A total of 12 NSCLC patients with EGFR-sensitive gene mutation treated by gefitinib from Xianyang Center Hospital of Shaanxi Province and Yongchuan Hospital of Chongqing Medical University from March 2015 to April 2019 were selected. Plasma samples before and after drug resistance were collected, and 6 samples in sensitive stage and 6 samples in drug-resistant stage were taken. Gene microarray was used to screen the differentially expressed lncRNA and mRNA; the biological pathway and the function of the differentially expressed mRNA were obtained by using the gene ontology (GO) function annotation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis.Results:The microarray detection results showed that the expression profiles of lncRNA and mRNA in the plasma of NSCLC patients were different before and after gefitinib-resistance. Fold change≥2 and P < 0.05 were taken as the differential gene screening standard, finally 38 differentially expressed lncRNAs and 53 differentially expressed mRNAs were found. Compared with the sensitive stage, 18 lncRNAs were differentially up-regulated and 20 lncRNAs were down-regulated in the drug-resistant stage; the largest up-regulation lncRNA was RP1-102K2.6 (fold change was 47.31), and the largest down-regulation lncRNA was RP11-149I2.4 (fold change was 24.34). In mRNA expression microarray, compared with sensitive stage, the expressions of 29 mRNAs were up-regulated and 24 mRNAs were down-regulated in the drug-resistant stage, the largest up-regulation mRNA was CUL2 (fold change was 58.49), the largest down-regulation mRNA was CHEK2 (fold change was 23.29). GO functional analysis showed that the differentially expressed mRNA in the plasma of patients with gefitinib-resistance were enriched in the apoptosis and protein binding regulation process. KEGG analysis showed that the differentially expressed mRNA mainly targeted cancer pathway, NSCLC pathway and other pathways. Conclusion:For NSCLC patients with EGFR gene sensitive mutation, there are multiple differentially expressed lncRNAs and mRNAs in plasma before and after drug resistance, and the differential expression may play an important role in the mechanism of gefitinib resistance.

OBJECTIVE: To study the effective method of comprehensive evaluation and analysis of hospital valuable medical equipment performance. METHODS: The operation performance of 6 valuable equipment was evaluated by cost-benefit method, comprehensive index method and public evaluation method. RESULTS: Utilize equipment information management methods for data collection and evaluation, and construct an assessment data model based on evaluation indicators from three aspects: equipment operation status, profitability status, and scientific research contribution. CONCLUSIONS Through the performance analysis of different types of valuable medical equipment, a more real and comprehensive quantitative analysis is carried out, which plays a key role in the reasonable purchase, efficient operation and avoiding idling.
Equipment and Supplies, Hospital ; Hospitals ; Data Collection ; Information Management ; Surgical Equipment

In October 2016, a male patient attacked by a black bear was treated in the Department of Plastic Surgery, the Second Affiliated Hospital of Kunming Medical University.The patient had facial skin and soft tissue defects, and zygomatic arch and buccal damage. The patient received three operations, including debridement, scapular free skin flap transplantation, and reconstruction of zygomatic arch. The facial appearance recovered well after 6-months follow-up.

OBJECTIVE: To explore the pathogenesis of two fetuses from one family affected with Joubert syndrome (JS). METHODS: Whole exome sequencing was employed to screen potential mutations in both fetuses. Suspected mutations were verified by Sanger sequencing. Impact of intronic mutations on DNA transcription was validated by cDNA analysis. RESULTS: Two novel TCTN1 mutations, c.342-8A>G and c.1494+1G>A, were identified in exons 2 and 12, respectively.cDNA analysis confirmed the pathogenic nature of both mutations with interference of normal splicing resulting in production of truncated proteins. CONCLUSION The genetic etiology of the family affected with JS has been identified.Above findings have enriched the mutation spectrum of TCTN1gene and facilitated understanding of the genotype-phenotype correlation of JS.
Abnormalities, Multiple ; diagnosis ; genetics ; Cerebellum ; abnormalities ; Eye Abnormalities ; diagnosis ; genetics ; Humans ; Kidney Diseases, Cystic ; diagnosis ; genetics ; Membrane Proteins ; genetics ; Mutation ; Pedigree ; Retina ; abnormalities ; Whole Exome Sequencing

Objective To construct a delirium risk prediction model for patients in the intensive care unit(ICU)based on the related psychological risk factors of delirium,and provide a new idea for the identification of delirium in ICU patients.Methods A prospective observational study method was conducted.From September 2019 to September 2020,a total of 165 patients in the department of ICU of the Second Affiliated Hospital of Chongqing Medical University were selected as the research objects.The patients were assessed by general information questionnaire,eysenck personality questionnaire-revised,short scale for Chinese(EPQ-RSC),state-trait anxiety inventory(STAI),Hamilton depression scale(HAMD),trait coping style questionnaire(TCSQ),and confusion assessment method of ICU(CAM-ICU).The binary Logistic regression model was used to explore the risk factors of delirium in ICU patients,and a nomogram model was constructed to verify the accuracy of the model.Results After excluding 7 cases of invalid data,158 patients were finally included,23 were believed to be suffering from delirium,and the incidence of delirium was 14.56%.The univariate analysis showed that compared with the non-delirium group,the age of patients was significantly increased(years:72.91±6.75 vs.63.36±10.14),the proportion of patients with history of alcoholism,cognitive impairment and mechanical ventilation in the delirium group was significantly increased[history of alcoholism:17.4%(4/23)vs.5.2%(7/135),history of cognitive impairment:30.4%(7/23)vs.5.2%(7/135),history of mechanical ventilation:78.3%(18/23)vs.40.7%(55/135),all P<0.05],the length of ICU stay was significantly prolonged(days:7.26±1.66 vs.4.93±2.15),the neuroticism score(7.78±2.66 vs.5.07±2.77),the negative coping score(30.70±6.54 vs.25.76±5.41),the HAMD depression score(15.04±4.55 vs.10.76±3.77),and the trait anxiety score(49.48±7.14 vs.44.10±8.66)were significantly increased(all P<0.05).Logistic regression analysis showed that age,neuroticism score,HAMD depression score,trait anxiety score,the length of ICU stay,and history of mechanical ventilation were all risk factors for delirium in ICU patients[odds ratio(OR)value and 95%confidence interval(95%CI)were 1.11(1.02-1.22),1.50(1.13-1.99),1.39(1.15-1.69),1.13(1.03-1.25),1.47(1.04-2.06),6.52(1.19-35.73),P values were 0.02,0.01,0.01,0.01,0.03,0.03,respectively].A nomogram model was constructed based on the delirium risk factors,area under the curve(AUC)=0.96 and 95%CI was 0.93-0.99,the Youden index was 0.87,the sensitivity was 100%,and the specificity was 87%.The results of the Hosmer-Lemeshow goodness of fit test suggested thatχ2=5.13,P=0.74,suggesting that the prediction model had good discrimination.Conclusion This study constructed a risk prediction model for delirium in ICU patients based on neuroticism,depression,trait anxiety and other factors,and the result showed that the model had good discrimination and accuracy,offering a new method for identifying ICU patients at high risk of delirium.