Objective To establish fully automated sample pooling, nucleic acid extraction, amplification and detection method for HBV DNA testing, and investigate the seroconversion and genotype in HBV DNA positive donors. Methods Individual donor plasma samples serologically negative for HBV were pooled by STAR2000 sampling processor with a size of 24. Nucleic acid were automatically extracted by MPLC simultaneously, amplified and detected by Roche COBAS AMPLICOR system. The sensitivity of detection was determined by international standard. HBV DNA positive donors were genotyped and followed up by serological tests. Results The 95% detection limit for this automated HBV DNA testing system was 38.9IU/ml,with 95% confidence interval (21323),eight out of 16512 specimens were PCR positive for HBV DNA,with a positive rate of 0.049%. Three of the 8 DNA positive donors were genotype C,2 genotype B, 1 genotype D,and the other 2 uncertain。Six of the eight HBV DNA positive donors were followed up, and three of them seroconverted。 Conclusion Fully automated HBV DNA detection method can be applied in blood screening,and will further increase the safety of blood supply.

BACKGROUND:It is a great potential study that calcium sulfate product loaded with antibiotics is developed, but this product is not systematicaly studied and its biocompatibility and security need to be further studied.
OBJECTIVE:To evaluate the biocompatibility and safety of vancomycin- or gentamicin-loaded calcium sulfate bone.
METHODS: (1) Hemolysis test: vancomycin-loaded, gentamicin-loaded calcium sulfate extracts, double distiled water and saline were added into rabbit anticoagulant blood samples. (2) Micronucleus test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts, cyclophosphamide and normal saline solution were intraperitonealy injected to mice, respectively. (3) Acute toxicity test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts, and normal saline solution were intraperitonealy injected to mice, respectively. (4) Pyrogen test: the mice were injected vancomycin-loaded and gentamicin-loaded calcium sulfate extractsvia the ear vein. (5) Intradermal stimulation test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts were respectively injected into the unilateral spine of rabbits, respectively. (6) Intramuscular implantation test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts were respectively injected to the dorsal muscle of rabbits. (7) Intraosseous implantation test: vancomycin-loaded and gentamicin-loaded calcium sulfate were implanted into the necrotic femoral bone of rabbits.
RESULTS AND CONCLUSION:Both vancomycin-loaded and gentamicin-loaded calcium sulfate products, which have no hemolytic reaction, genetic toxicity, acute toxicity, pyrogen reaction and skin irritation, are considered to have good biocompatibility and safety.

Aim To explore the mechanisms of action (MOA) of synergistic anticancer function in the combination of berberine and evodiamine.Methods We first analyzed the action of suppression in the drug combination from the cell level and validated the dose scope as well as ratio of concentration in synergistic effects of drug combination.Then, the miRNA chip of liver cancer cell BEL-7402 under different treatment was analyzed.By building the miRNA-mRNA network, the MOA of the synergistic drug combination was illustrated.Results Berberine and evodiamine used in combination could significantly synergistically suppress the proliferative ability of liver cancer cells.The special differentially expressed miRNAs (DEmiRNAs) mainly participated in some cancer proliferation-related pathways and biological processes, such as MAPK signaling pathway, endocytosis pathway and insulin signaling pathway.The special target genes influenced by the drug combination not only covered three kinds of membrane receptors, but also took part in the regulation of downstream pathways.Conclusions From the regulation of miRNAs, it is clear that berberine may play a primary role in the synergistical suppression activity of the drug combination in cancer cells.The discovery of synergistic MOA in the combination of berberine and evodiamine from the miRNA level will provide a new guidance to explore more synergistic drug combinations in the future.

Objective To study the prevalence and sequence homology of virulence genes exoU and exoS in 53 strains of pan-drug-resistant Pseudomonas aeruginosa .Methods The virulence genes exoU and exoS were detected by PCR.Sequence homo-logy was analyzed by BOX-PCR.Results Of the 53 clinical isolates of pandrug-resistant Pseudomonas aeruginosa ,the exoS+/exoU- genotype was identified in 40 strains,exoU+/exoS - genotype in 10 strains,exoS +/exoU+ genotype in 1 strain, and exoS-/exoU- genotype in 2 strains.BOX-PCR results showed that 41 exoS+ isolates belonged to 24 genotypes,and 11 exoU+ strains could be grouped into 7 genotypes.Conclusions The prevalence of virulence genes is high in clinical isolates of pandrug-resistant Pseudomonas aeruginosa .BOX-PCR fingerprint analysis combined with sequence homology analysis is help-ful for effective monitoring and control of hospital pandrug-resistant pseudomonas aeruginosa infection.

Objective To observe the change of peripheral blood CD34+ cell level in patients with acute cerebral infarction, and explore its relationships with cerebrovascular risk factors,neurological function and carotid artery intima-media thickness (IMT). Methods The 45 patients with acute cerebral infarction (onset within 72 h) (infarction group) and 27 patients with cerebr ovascular risk factors but without cerebral infarction (high-risk group) were chosen for the study. The cerebrovascular disease risk factors including history of alcohol abuse, smoking, coronary heart disease, hypertension, diabetes, abnormal levels of serum triglycerides, total cholesterol,low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were recorded in all subjects. The peripheral blood CD34+ cell levels were measured by flow cytometry.The correlations of peripheral blood CD34+ cell level with cerebrovascular disease risk factors were analyzed. The neurological function and carotid artery IMT were recorded in infarction group, and the correlations of peripheral blood CD34+ cell level with neurological function and carotid artery IMT were analyzed. Results (1) The peripheral blood CD34+ cell level was significantly negatively correlated with coronary heart disease, hypertension, diabetes and LDL-C level (r =- 0. 749,-0. 717, - 0. 688, - 0. 764, all P＜0. 01) ; (2) Multiple linear regression analysis showed that peripheral blood CD34+ cell level was an independent relative factor of acute cerebral infarction (P＜0.05); (3) The peripheral blood CD34+ cell level was lower in infarction group than in high-risk group, and was significantly negatively correlated with neurological deficit score (r=-0. 721, P＜0.01) and carotid artery IMT (r= -0. 695, P＜0. 01). Conclusions Peripheral blood CD34+ cell level could be an independent relative factor of acute cerebral infarction; The peripheral blood CD34+ cell level is significantly negatively correlated with neurological function and carotid artery IMT in patients with acute cerebral infarction; And it can be used as cytological marker which reflect early vascular endothelial function in patients with ischemic stroke.

Objective:To evaluate the association between brachial-ankle pulse wave velocity ( ba-PWV) and metabolic syndrome ( MetS) among the Chinese elderly and the gender difference .Methods:We conducted a population-based cross-sectional study in a representative urban area of Beijing , China. A sample of 2 102 community elderly (848 males, and 1 254 females) aged 60 to 95 years were included in the study .MetS was defined according to the 2009 harmonizing definition .Results:The prevalence of MetS was 59.1% (50.1% in males and 65.2% in females, P<0.001).The baPWV value was be-tween 8.2 to 45.6 (20.0 ±4.4) m/s, and showed an increasing trend with age (P<0.001).The par-tial correlation showed baPWV was positively associated with BMI (r =0.076, P =0.037), systolic blood pressure (r=0.380, P<0.001), diastolic blood pressure (r=0.276, P<0.001), triglyceride (r=0.040, P=0.046), fasting blood glucose (r=0.140, P<0.001), 2-hour post-meal blood glu-cose (r=0.121, P<0.001), and negatively associated with HDL-C (r=-0.128, P=0.048).There was a strong association between baPWV and prevalence of MetS and its component number in females but not in males .Compared with the lowest quartile of baPWV , the adjusted ORs were 1 .22 ( 95%CI 0.83-1.79), 1.32(95%CI 0.90-1.93), 1.46(95%CI 1.00-2.14) in males and 1.28(95%CI 0.93-1.77), 1.55(95%CI 1.12 -2.16), 1.86(95%CI 1.32-2.61) in females for the second, third and top quartiles .Conclusion:The prevalence of MetS increases substantially with increasing levels of baPWV among the Chinese elderly , especially in females .

Objective Our purpose was to evaluate the teaching satisfaction of clinical epi-demiology among medical postgraduate and to come up with measures for further improvement of teaching quality. Methods A self-administered questionnalre was given to all the medical postgradu-ates and doctoral students of Grade 2013 by cluster sampling when they finished the course of clinical epidemiology. A total of 559 graduate students, including 324 graduate students (58%), 235 doctoral students (42%), recruitment graduate students 350 (62.6%), on-the-job graduate student 209 (37.4%).The contents of the questionnalre included many aspects such as the investigation object in general, teaching materials evaluation satisfaction, curriculum and teacher satisfaction evaluation. Respondents ' self-administered manner was adopted. Parallel input was done by using Epidata software; data were analyzed with SPSS 19.0 software, continuous variable was made by x±s, categorical variables was expressed by n(%). Continuous variables were compared among groups by t test and analysis of vari-ance. Classification grouping variable was compared by chi-square test, and P<0.05 for the difference was statistically significant . Results Overall evaluation of theory was below that of the internship teaching materials. The satisfaction rate of practicability, meeting the learning needs and difficulty degree for theory and practice teaching material were 83.7% (468/559), 87.5% (489/559), 67.1% (375/559) and 92.7%(518/559), 89.6%(501/559), 83.0%(464/559) respectively. 41.6%(87/209) of on-the-job students and 36.7% (119/324) of postgraduates considered theory teaching material was difficult for them. The satisfaction rate for faculty teaching attitude (99.5%, 556/559), faculty teaching method (98.6%,551/559), the theories combining with practice teaching method (97.5%, 545/559) were high, but the satisfaction rate for teaching hours was low (67.4%, 377/559), with one third students consider more hours for the course. Conclusion The Teaching effect of clinical epidemiology is falrly good in our school. The postgraduates are satisfied with the teaching materials, curriculum setting, and teach-ing faculty. But there are also some shortcomings. In the future teaching work, we should appropriately increase the hours, and set corresponding teaching contents and methods according to the different learning characteristics of the on-the-job graduate students and recruitment graduate students, to improve the teaching effect and teaching quality.

Objective To establish the entirely automatic method of nucleic acid amplification testing (NAT) for blood screening and to study the feasibility of NAT.Methods Using entirely automated extraction method to extract nucleic acid , amplified and detected by Roche COBAS AMPLICOR system,evaluating the sensitivity and efficacy.Results The 95% limits of HBV DNA, HCV RNA/HIV-1 RNA tests by automation system were 38.9,16.4IU/ml and 20.4 copies/ml,95% Confidence Intervals were [21,323], [10.5,342] and [12,300] respectively.8 of 16 512 donations were PCR for HBV DNA positive,the DNA positive rate was 0.048%.7/8 donations were Anti-HBc positive,The last one was also converted positive.No positive HCV RNA and HIV RNA was detected. 3/6 following up samples seroconverted.Conclusions The entirely automatic system can be applied in blood screening.

Objective:To investigate the correlation between variations in mitochondrial DNA (mtDNA) control region (D-loop) and keloids.Methods:A total of 216 patients with keloids were collected from Department of Dermatology, the First Affiliated Hospital of Kunming Medical University from 2016 to 2019. Total DNA was extracted from peripheral blood samples of all the patients, as well as keloid tissues and perilesional normal skin tissues of 25 patients with keloids. Peripheral blood samples were collected from 299 health checkup examinees without keloids in Health Examination Center, the Affiliated Hospital of Yunnan University, who served as controls. PCR amplification and Sanger sequencing were performed on the mtDNA D-loop region, and mutation sites in each sample were analyzed by comparisons with the revised Cambridge Reference Sequence (rCRS) . Haplogroups were assigned in the 2 groups by using Phylotree-mtDNA tree Build 17. Mutations in the mtDNA D-loop region were compared among keloid tissues, perilesional normal skin tissues and peripheral blood samples. A median-joining network was constructed via network 5.0 software. Binary logistic regression analysis was performed to investigate the correlation between haplogroup frequencies and the occurrence of keloids, and chi-square, t and t′ tests were used to analyze clinical data. Results:Among the 216 patients with keloids, variations in mtDNA D-loop region were classified into 10 haplogroups, including A, B, D, R9, G, M*, M7, M8, M9 and N9, with the haplogroups R9 and M9 showing the highest (21.3%, 46/216) and lowest (0.9%, 2/216) frequencies respectively. The frequencies of haplogroups M7 ( P=0.040, OR=0.248, 95% CI: 0.066 - 0.937) and N9 ( P=0.048, OR=0.191, 95% CI: 0.037-0.986) were significantly lower in the patients with keloids than in the controls. The median-joining network plot showed that the distribution pattern of the haplogroup M7 differed between the patients with keloids and controls. Significantly less number of lesional sites and younger age of onset were observed in the patients with haplogroup M7 compared with those with non-M7 haplogroups ( P=0.000 1, 0.045, respectively) . Conclusion:The haplogroup M7 is correlated with the occurrence of keloids, and may be a potential protective factor for keloid formation.

Objective:To analyze the clinical phenotypes and prenatal diagnosis of a pedigree with oculo-facio-cardio-dental (OFCD) syndrome.Methods:A pregnant woman at 17 gestational weeks was admitted to the Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School in 2017 for genetic counseling. Genetic tests as performed for the proband (the pregnant woman), her husband, and the induced fetus of last pregnancy genetic test and the detected variants were analyzed and verified by chromosomal microarray analysis (CMA), multiplex ligation-dependent probe amplification (MLPA) and quantitative real time-polymerase chain reaction (Q-PCR). The detection platform established by MLPA and Q-PCR technology was used to perform prenatal diagnosis of the present pregnancy. Other family members were screened for BCOR gene mutation. Related mutation types were retrieved from ClinVar database with term of " BCOR", and related literature from CNKI and PubMed with terms of "OFCD syndrome", " BCOR gene", and "oculo facio cardiac dental syndrome" to summarize the clinical manifestations, mutation type and pathogenesis of this disease. Results:The proband has congenital cataracts, long face, congenital atrial septal defect, and severe dental malformations, which were consistent with the clinical features of OFCD syndrome. WES suggested that the proband and her induced fetus were suspected of having a large submicroscopic deletion of the exons of BCOR gene, which was confirmed by CMA, MLPA and Q-PCR, with a 105 kb deletion containing BCOR exons 1-15. The amniotic fluid genetic analysis of the present pregnancy showed that the fetus has a normal female karyotype, and did not carry the same BCOR gene copy number abnormality as the proband. The child grew and normally developed without any characteristic manifestations of OFCD syndrome during follow-up. Other families of the proband did not show clinical features of OFCD syndrone, and no BCOR gene copy number abnormality was detected. A total of 35 cases of BCOR gene mutation types related to OFCD syndrome were retrieved from ClinVar database. The data analysis revealed that the differences in clinical manifestations between Lenz microphthalmos syndrome and OFCD syndrome were mainly caused by different mutation types of BCOR gene. Among the 90 retrieved cases of OFCD syndrome obtained through literature, only one case was reported in China. Analysis of these 90 cases showed that the characteristic manifestations of OFCD syndrome, involving the eye, face, heart, teeth, and skeletal system. OFCD syndrome were confirmed in the proband and her induced fetus according to the clinical manifestation and the mutation type of BCOR gene. Conclusions:The clinical manifestations of OFCD syndrome are complicated, caused by various mutation types of BCOR. Systematic molecular genetic technology can be effectively applied for gene and prenatal diagnosis of OFCD syndrome.