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.

Cytochrome P 4501 A 1, Cytochrome P 4501 B 1,Vascular endothelial growth factorand carbonic anhydrase Ⅸ belong to the downstream genes of aryl hydrocarbon receptor (AhR)and hypoxia inducible factor 1(HIF-1)signaling pathways. The abnormal expression of those genes were regarded to associated with the occurrence,development and angiogenesis of lung cancer. In this paper, the relationship between CYP1 A 1,CYP1 B 1,VEGF,CAⅨgenes and lung cancer was summarized, which aims to provide new ideas for lung cancer research.

Objective To evaluate the antitumor effects of interferon (IFN)γ-endostatin based gene radiotherapy in a metastatic breast tumor model of mice, and to elucidate the possible mechanisms involved. Methods Murine mammary adenocarcinoma 4T1 cells transfected with pEgr-IFN-γ and pEgrendostatin plasmids were irradiated with 2-20 Gy of X-rays. IFN-γ and endostatin levels in the culture supernatants were measured. Female BALB/c mice were inoculated with 1 × 105 of 4T1 cells by mammary fat pad injection, and divided randomly into control, empty vector, gene therapy (pEgr-IFN-γ and pEgrendostatin), radiotherapy, and combined gene-radiotherapy groups. Tumor/body weight ratio, lung metastases, and survival of the tumor-bearing mice were observed. Splenic cytotoxic T-lymphocyte (CTL)and natural killer (NK) cell activity and intratumor microvessel density were also assessed. Results Irradiation significantly enhanced the section of IFN-γ and endostatin from the transfected 4T1 cells.Compared with gene therapy or radiotherapy alone, combined gene-radiotherapy resulted in the maximal attenuation in tumor growth rate, lung metastases and increased survival. The activities of CTL and NK cells were significantly enhanced and intratumor microvessel density reduced ( t = 2. 120-22.140, P ＜ 0.05 ).Conclusions IFN-γ-endostatin-based gene-radiotherapy could provide a potential antitumor effect in a murine metastatic breast tumor model, which may be related to IFN-γ-stimulated CTL and NK cell activation, and endostatin-induced antiangiogenic activity. Gene-radiotherapy could serve as a neoadjuvant therapy for the locally advanced breast cancer.

Due to the advantages of being unaffected by fetal gender, ease of detection, and good stability, circulating cell-free fetal RNA (cffRNA) is a potential biomarker in obstetric practice. Current evidence has shown that placenta is the main source of circulating cffRNA. In view of the abnormal expression levels in women with preeclampsia and intrauterine growth restriction, circulating cffRNA is proposed as a potential tool to predict or diagnose these diseases. A summary of the molecular characteristics and the applications in preeclampsia of circulating cffRNA is reviewed, in order to evaluate the hypothesis for the prediction of preeclampsia by cffRNA.

Objective:To understand the possible detected mosaicism chromosome karyotyping using uncultured chorionic villus samples.Methods:This study retrospectively analyzed the clinical data of singleton pregnant women who underwent fetal chromosome karyotyping of uncultured chorionic villus samples at the Prenatal Diagnosis Center of Tianjin Central Hospital of Gynecology and Obstetrics from January 2016 to January 2019. Prenatal diagnosis indicators, fetal karyotypes, the incidence of chromosomal mosaicism and subsequent diagnosis, and perinatal outcomes were analyzed. Amniocentesis was performed when chromosomal mosaicism was identified. Descriptive statistical analysis was used for data analysis.Results:(1) A total of 438 pregnant women with available follow-up data were enrolled. Increased nuchal translucency (56.6%, 248/438) was the major indication for prenatal diagnosis. The karyotype analysis indicated that 79.5% (348/438) were normal, and 2.7% (12/428) were mosaicism. (2) Of the 438 cases, 336 cases (76.7%) were delivered at term, of which 327 cases were uncomplicated. There was one case of premature rupture of membranes within one week after amniocentesis and eight cases of abortion/fetal death between one week after the amniocentesis and 28 weeks of gestation. Of these nine cases, four had chromosomal abnormalities, and five had normal karyotypes. Termination of pregnancy was selected in 65 cases (14.8%) and 28 cases (6.4%) delivered before term. (3) Among the 12 (2.7%) cases of chromosomal mosaicism verified by fetal karyotyping through amniocentesis, four were confined placental mosaicism; six were abnormal chromosomal karyotypes in chorionic villous and amniotic fluid; one was true fetal mosaicism; one was a false positive. Among the 12 cases, three continued to term, one was preterm delivered, and eight selected labor induction, including three cases each of trisomy-21 and ultrasonographic structure abnormalities, and one case each of fetal growth restriction and labor induction based on patient preference.Conclusions:Karyotype analysis of uncultured chorionic villus samples may detect a certain proportion of mosaicism. Therefore, combining fluorescence in situ hybridization to achieve an accurate diagnosis and a detailed and systematic ultrasonic scan are recommended.

Objective:To investigate the effects of moxibustion on the serum metabolism in healthy human body based on the 1H nuclear magnetic resonance (1H NMR) metabolomics technology, and to find the differences in metabolites, as well as to elucidate the effects of moxibustion on healthy human body from the viewpoint of global metabolism. Methods:Sixty subjects of healthy young men from the enrolled students were randomly divided into a moxibustion group and a control group using random number table, with 30 cases in each group. Subjects in the moxibustion group accepted mild moxibustion on the right Zusanli (ST 36), once a day, 15 min for each time, and continuous treatment for 10 d; those in the control group did not receive any intervention. There were 28 cases in the moxibustion group and 23 cases in the control group after interventions. On the 1st day, 5th day and 10th day of the intervention, serum samples were collected from subjects of the two groups, and metabolic spectra were obtained by the1H NMR technology. Results: Before and after the intervention, serum1H NMR of the moxibustion group was significantly different, while the difference was insignificant in the control group. Metabolite changes in the moxibustion group were mainly in low density lipoprotein (LDL)/very low density lipoprotein (VLDL), valine, isoleucine, leucine, lactic acid, glutamine, citric acid, polyunsaturated fatty acids, creatine, glycine, glycerol, glucose, tyrosine, histidine, formic acid, alanine, lysine, acetic acid, and glutamic acid. Conclusion:Moxibustion can cause changes of serum metabolic patterns in healthy human by influencing the concentrations of branched-chain amino acids, polyunsaturated fatty acids, and other metabolites to strengthen body's metabolisms of amino acids and fatty acid.

With an increasing proportion of sedentary lifestyle related chronic diseases, an efficient way to help fight the growing world epidemic of non-communicable diseases (NCDs) and promote public health in a fast-changing world is urgently needed. Exercise and medicine science have played an important role in prevention and control of NCDs. However, the current status and future development directions regarding the integration of exercise science and medicine (IESM) in China have not been well discussed. During 2017 Belt and Road Initiative Global Health International Congress & 2017 Chinese Preventive Medicine Association-Chinese Society on Global Health Annual Meeting, Xi'an Jiaotong University Global Health Institute, and the Integration of Sport and Medicine Innovation Research Center, China Institute of Sport Science organized the IESM Forum on September 27. Six leaders and experts from China and abroad presented excellent research reports in the IESM Forum from several aspects. Experts reached a consensus at the end of their discussions that, in the near future, the IESM will be a key part of a sustainable health care system and as such should contribute to prosperity in China and around the world. However, Abstract compared with European, American and Japanese universities, the development of the IESM in China is in its early stages with huge development potential.

Objective To explore the effect of posterior debridement, grafting and internal fixation for treatment of non-specific lumbar intervertebral infection. Methods Clinical data of 20 patients with non-specific lumbar intervertebral infection treated in General Hospital of Ningxia Medical University from October 2013 to June 2013 were retrospectively analyzed. There were 15 males and 5 females with an average age of 41 years (range, 36-51 years). All patients suffered from single lumbar inter-vertebral infection, including 3 cases at L2/3,4 at L3/4,10 at L4/5 and 3 at L5/S1. All 20 cases underwent one-stage posterior debride-ment, autogenous bone grafting and internal fixation, tissue samples in focus were collected for bacterial culture and pathological examination. The disease controlling statues were evaluated based on laboratory results of ESR and CRP. Imaging examinations were taken to evaluate the fusion of vertebral body. Clinical effects were evaluated using the visual analog scale (VAS) and the Jap-anese Orthopaedic Association scores (JOA) score of lumbar fumction. Results All patients underwent the surgery successfully. The surgery duration time was 90-160 min, average 125 min, and the blood loss was 200-700 ml, average 360 ml. Cerebrospinal fluid leakage occurred in one case. Postoperatively, all patients experienced significant reliefof back pain, improving in the func-tion of movement, and no fever. The lower back VAS score: average (5.35 ± 1.15) points before operation , average (2.76 ± 0.34) points one week after operation, and an average score of (0.85±0.65) points by the last follow-up time. JOA lumbar function score:all patients were effective after operation, the improvement rate was excellent in 65%(13cases), good in 25%(5 cases), and pass-able in 10%(2cases). Comparing with preoperation, the excellent and good rate was 90%. All patients ESR and CRP returned to normal levels at the last follow-up. Ordinary bacterial culture was positive in 8 cases and negative in 12 cases. The pathogens iden-tified were staphylococcus aureus (6 cases), Escherichia coli (2 cases) and staphylococcus epidermidis (2cases). All incisions achieved primary healing. All patients were followed up from 6-18 months (average,12 months), and the symptom of pain relieved significantly. No recurrent infection had happened. A solid bony fusion was found in all patients at 6-14 months (average, 8.5 months) after the surgery. Conclusion Posterior debridement, grafting and internal fixation are effective treatments for non-spe-cific lumbar intervertebral infection, can reduce the time of staying in hospital, this operation is safe and reliable.

Objective Investigate the effect of neonatal hyperbilirubinemia on the detection of HBsAg by chemiluminescence and its elimination methods.Methods Case control study.The HBsAg in human serum was detected in 200 cases of hyperbilirubinemia neonates who were hospitalized in Beijing Children's Hospital,Capital Medical University from July 2015 to May 2016 and whose serum total bilirubin level exceeded 200 μmol/L.The positive serum was further detected by 16 200×g high-spoed centrifugation or blue light irradiation for 8 hours,and the results of re-assay of HBsAg were recorded.The retest positive serum wastested for HBV DNA load and checked the results of their mother's examination in HBV.136 adult serum samples with total bilirubin levels exceeding 200 μmol/L in the Peking University First Hospital,were taken as reference to compare the influence of hyperbilirubinemia between adults and newborns on the determination of HBsAg.Results The median level of serum total bilirubin in neonates was 259.0 μ mol/L (226.5,312.5);median level of indirect bilirubin 244.1 μmol / L(212.5,295.8).Median level of serum total bilirubin in adults 356.4 μmol/L(295.9,435.1);median level ofindirect bilirubin 137.1 μmol/L (107.8,172.7).The HBsAg test was negative in adults,11 cases (5.5％) were positive in newborns,their" HBV DNA load was less than＜100 IU/ml.Among them,9 have inoculated hepatitis B vaccine and 2 were unknown.10 of 11 mothers of infants were healthy and 1 was positive for HBsAg,HBeAb,HBcAb.2 of the 11 positive specimens turned negative of HBsAg after high-speed centrifugation.In addition to high speed centrifugation,4 cases turned negative after blue light irradiation.5 cases remained positive after high speed centrifugation and blue light irradiation.Conclusions Neonatal hyperbilirubinemia,which is different from that of adults,is mainly caused by indirectly bilirubin increased,which is one of the main reasons for false positive detection of HBsAg by chemiluminescence in neonates.High-speed centrifugation and blue light irradiation can eliminate the influence of serum indirect bilirubin on the detection of HBsAg to the greatest extent.

Objective:To inhibit the Itch gene expression of T-lymphocytes and investigate the immune activity of T lymphocytes using ultrasound-targeted microbubble destruction (UTMD).Methods:T lymphocytes were separated by magnetic bead, and to establish an Itch gene targeted short hairpin RNA (shRNA) plasmid. There were three groups in this study: ①experimental group, Itch-shRNA plasmid-SonoVue microbubbles combined with ultrasound irradiation; ②control group, negative control shRNA plasmid-SonoVue microbubbles combined with ultrasound irradiation; ③blank group, untreated. Forty-eight hours after UTMD transfection, transfection efficiency was detected and analyzed by fluorescence microscopy and flow cytometry, the expression of Itch protein was measured with Western blotting.Seventy-two hours after UTMD transfection, the secretion of interleukin 2 (IL-2) and interferon γ (IFN-γ) in the cell supernatant were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA was used to compare the differences between groups, and LSD- t test was used to compare the differences between groups. Results:The UTMD mediated shRNA transfection rate was 52.3%, and the relative expression levels of Itch protein in the experimental group, control group and blank group were 0.301±0.080, 0.773±0.101 and 0.719±0.090, respectively, and the difference was statistically significant( F=24.441, P<0.01). The Itch gene expression can be effectively suppressed in the experimental group. Seventy-two hours after transfection, the concentrations of IL-2 in the experimental group, control group and blank group were (417.3±37.1)ng/L, (158.7±17.3)ng/L and (147.0±10.2)ng/L, respectively, and the difference was statistically significant( F=118.701, P<0.001) and the concentrations of IFN-γ were (168.3±12.1)ng/L, (74.3±3.7)ng/L and (74.6±7.1)ng/L, respectively, and the difference was statistically significant ( F=126.833, P<0.001). The expression levels of IL-2 and IFN-γ in the experimental group were significantly higher than those in the control group and the blank group. Conclusions:UTMD mediated shRNA transfection can significantly decrease the expression of Itch and promote immune activity of T lymphocyte.