Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid (CSF) were recruited and patients with craniocerebral trauma were recruited as controls.Standard plasmids were constructed using standard CN strain.Real time fluorescent quantitative polymerase chain reaction (RT-FQ-PCR) was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis,which were compared with the results of CSF ink staining,fungal culture and antigen detection.The diagnostic values of single gene test and combined gene test were compared by chi square test.Results Among 23 CN meningitis patients,22 (95.6％) were CAP10 mRNA positive detected by RT-FQ-PCR,which was significantly higher than both ink staining (16/23,69.6％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).There were also no statistical significant differences between combined detection of CAP 10 + VAD1 and CAP 10 or VAD1 single gene test (P＞0.05).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.

Objective Queuing theory is the mathematical study of how waiting lines or queues are formed and dissipated o -ver time, which aims to work out the optimal design and optimal control of queuing systems based on the research of probability and regularity of various queuing systems .The aim of paper was to find out the maximum acceptable waiting time for outpatients in stomatol -ogy department and explore the rational allocation of dentists based on queuing theory model . Methods Questionnaires , worktime measurement and queuing theory model were applied to calculate indicators of queuing system in outpatient services of Stomatology De -partment , getting the maximum waiting time accpetable for patients and the reasonable number of dentists . Results The maximum acceptable waiting time for outpatients was 34.02 ±7.07 minutes, and it was reasonable to allocate 25 doctors in the morning and 16 doctors in the afternoon . Conclusion Applying queuing theory helps to optimize dentist number in outpatient services of Stomatology Department and provide scientific reference to improve medical efficiency .

Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

OBJECTIVETo test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics.

METHODSPeripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR).

RESULTSThe traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups.

CONCLUSIONResults indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.

Adenoviridae ; genetics ; physiology ; Cells, Cultured ; Cytokines ; genetics ; immunology ; Dendritic Cells ; immunology ; virology ; Genetic Vectors ; genetics ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Humans ; Lymphocyte Culture Test, Mixed ; Recombinant Fusion Proteins ; genetics ; immunology

Objective To discuss training of the nurses in outpatient blood collection room with the digital training modular of blood sample collection to improve the quality of blood sample collection. Methods Nurses were trained with the digital training modular by multimedia,group discussion to impmve the quality of blood sample collection continuously. Results The unqualified blood sample rate in the same season after training were statistically different compared with that before training. Conclu-sions Training the nurses in outpatient blood collection room with the digital traimg modular of blood sample collection have actual direction value to improve the quality of blood sample collection.

Objective To study the prevalence of the mtDNA A1555G gene mutation in Chinese population with nonsyndromic hearing impairment.Methods PCR-RFLP,directional sequencing of PCR products were applied in 325 patients with nonsyndromic hearing impairment and 50 normal controls.Results The mutation rate of the mtDNA A1555G was 14.5% (47/325),28 of 47 cases were homozygosis,19 of 47 cases were heterozygosis.The same mutation was not detected in the control subjects.Conclusion The mutation rate of the mtDNA A1555G is relatively high in the Chinese NSHI patients,the mutation type includes both heterozygosis and homozygosis.

Objective To design and apply the digital training modular for nurses in blood sample collection in order to improve the quality of blood sample collection.Methods Based on the principle of evidence-based nursing,the records of unqualified blood samples in the clinical laboratory department were analyzed.Then,the modular of examination item list,tube choosing,volume of blood sample,patient preparation and theoretical foundation,influential factors of the quality of blood collection,and quality information were designed and applied to train nurses.Results After training,the unqualified rate of blood samples was significantly decreased from 0.56% to 0.34% (P<0.01).Conclusion The application of digital training modular of blood sample collection can improve the quality of blood collection.

Objective To develop a real time amplification refractory system(RT-ARMS-qPCR) quantitative PCR method with SYBR Green I to assess the mtDNA A1555G mutation. Methods A specific fragment flanking mtDNA 1555 site was amplified with PCR and ligated into a pGEM Easy T vector. Serial dilutions of the plasmid DNA were quantified the actual copy numbers were assessed using RF-ARMS-qPCR with SYBR Green I. RF-ARMS-qPCR was established with mismatched base pairs at 3' in the primer todetect the copy number of mtDNA containing wild or mutant mtDNA. The specificity of amplified products was checked by melting curve analysis. Results The intra- and interassay variation was 1.34% and 1.96%, respectively when the assay was used to detect 1 copy/ul recombinant template of plasmid. Thequantitative standard curve showed that the assay had good linear correlation from 102 copies/ul to108 copies/ul. This assay could be served for the quantification of other samples. There was significantcorrelation between frequency of mutant mtDNA and phenotype (r=0.771, P = 0.003) in hearing lossgroup. Conclusions The established assay can be used to detect quantitatively mtDNA A1555G mutation byRF-ARMS-qPCR. This assay is specific, stable and accurate. There is significant correlation betweenquantification of mtDNA AI555G and the severity of hearing loss.

Objective To explore the etiologies of fever of unknown origin(FUO)and methods for confirming di-agnosis in patients at a hospital,and provide reference for clinical diagnosis and treatment of FUO.Methods Pa-tients with FUO admitted to a hospital between January 2008 and July 2014 were performed clinical diagnosis with methods of serology,bacteriology,molecular biology,bone marrow aspiration,tissue biopsy,and diagnostic thera-py,the etiologies and final diagnosis of 224 patients were analyzed retrospectively.Results Of 224 FUO cases,189 (84.38%)eventually got confirmed diagnosis,35 (15.62%)were not confirmed.The percentage of infectious dis-eases,connective tissue diseases,malignant tumor,and other diseases were 50.45%,18.75%,9.82%,and 5.36%respectively.Among infectious diseases,the major pathogens were bacteria,followed by virus.The major connec-tive tissue diseases were systemic lupus erythematosus and polyarteritis nodosa;the main malignant tumor was he-matological tumor,lymphoma was the main form.Among 189 patients with confirmed diagnosis,30.16% and 24.34% were performed pathogenic and pathologic detection respectively,and 20.11% were performed the other (compre-hensive)methods.Conclusion Infectious diseases,connective tissue diseases,and tumor are major etiologies of FUO.

Objective To explore the clinical value of prenatal ultrasonography in the differentiation among the etiologies of fetal megacystis.Methods Twenty seven fetuses,diagnosed as fetal megacystis by prenatal ultrasonography,were retrospectively analyzed.The etiologies of fetal megacystis were presumed by such characteristics as keyhole sign,thickness of the bladder wall,amniotic fluid index,fetal sex and other combined signs.All fetuses were followed up until to the induction of labor or birth.Results Twenty seven singleton fetuses (19 males and 8 females) were diagnosed as megacystis.According to the characteristics and other combined signs,8 cases of posterior urethral valves (PUV),1 of prune belly syndrome(PBS),1 of megacystis-microcolon intestinal hypoperistalsis syndrome(MMIHS),1 of urethral atresia and 5 of chromosomal abnormality were presumed by prenatal ultrasound.Multiple malformations were found in 5 fetuses and there were also 6 fetuses with unknown reason originally.Among the 27 fetuses,21 were induced labor and 6 continued pregnancy to birth.Except for the 6 cases of unknown reason,etiologies of 17 fetuses with megacystis were confirmed by autopsy,genetic tests,surgery or further examination after birth.The accuracy rate of prenatal ultrasonography in the differentiation among the etiologies of fetal megacystis was 80.95％ (17/21).Conclusions On the basis of detailed prenatal ultrasonography and typical characteristics,it is reliable to differentiate the etiologies of fetal megacystis.Sometimes fetal megacystis may be one part of multiple malformations or complex syndrome,such as VACTERL syndrome.However,it is difficult for ultrasonography to diagnose vesicoureteral reflux(VUR)prenatally.