Objective To establish TaqMan probe real‐time Fluorescent Quantitative PCR in detecting Cryptococcus neoformans genomic DNA ,and to provide important method for detection of cryptococcal meningitis .Methods According to the Cryptococcus neoformans ITS‐rDNA sequences obtained from NCBI ,specific primers and probe were designed based on the conserved sequences , a specific 114 bp fragment was amplified by primers and probe ,then recombinant plasmid was constructed .Eight different concen‐trations from 1 .42 × 10 copy/μL to 1 .42 × 10 copy/μL were used as templates by 2 μL .In the optimum reaction condition ,the sen‐sitivity ,specificity and repeatability were evaluated and standard curve was established .15 clinical cryptococcal meningitis strains i‐solated from clinical diagnosis patients were detected .Results The real‐time PCR showed high sensitivity and specificity and was a‐ble to detect 2 .84 × 102 copies of plasmid DNA .The detection sensitivity was 2 .84 × 102 copies plasmid DNA by real‐time PCR ,no amplification curve was detected with human genomic DNA ,other fungus ,bacterias and viruses .The CV of inter‐assay were 2 .86% ,1 .48% and 1 .36% respectively with excellent reproducibility .Fifteen clinical isolated strains could be detected accurately . Conclusion A method of detection of Cryptococcus neoformans DNA by real‐time PCR is established successfully with high sensi‐tivity and specificity ,and the results are stable ,could diagnose cryptococcal meningitis rapidly and early .

Objective To explore the feasibility of measurement uncertainty in complete blood count using internal quality con-trol(IQC) data associated with Sysmex Network Communication System(SNCS) comparative data .Methods Complete blood count assay including white blood cell(WBC) count ,red blood cell(RBC) count ,hemoglobin(Hb) determination ,hematocrit(HCT) values and platelet(PLT) count were involved to evaluate measurement uncertainty according to the instruction of CNAS-TRL-001 .Meas-urement uncertainty evaluation was established by internal measurement reproducibility using IQC data ,and standard measurement uncertainty by bias using proficiency testing(PT) data and SNCS data ,followed by relative combined standard measurement uncer-tainty and relative expanded measurement uncertainty was calculated .Meanwhile ,the measurement uncertainty was compared by u-sing PT data and SNAS comparative data .Results Relative expanded measurement uncertainty of the above mentioned index by u-sing IQC data associated with PT data was the following :WBC(10 .02% ,7 .24% ,7 .04% ,from level 1 to level 3 respectively) ,RBC (2 .40% ,1 .72% ,1 .92% ,from level 1 to level 3 respectively) ,Hb(3 .54% ,2 .56% ,2 .50% ,from level 1 to level 3 respectively) , HCT(4 .12% ,3 .18% ,2 .86% ,from level 1 to level 3 respectively) ,PLT(15 .36% ,8 .86% ,7 .94% ,from level 1 to level 3 respec-tively) .Relative expanded measurement uncertainty of the above mentioned index by using IQC data associated with SNCS compar-ative data was the following :WBC(11 .66% ,7 .34% ,6 .40% ,from level 1 to level 3 respectively) ,RBC(2 .26% ,1 .60% ,1 .64%from level 1 to level 3 respectively) ,Hb(3 .36% ,2 .36% ,2 .10% ,from level 1 to level 3 respectively) ,HCT (3 .36% ,3 .04% , 3 .18% ,from level 1 to level 3 respectively) ,PLT (13 .34% ,8 .36% ,7 .14% ,from level 1 to level 3 respectively) .Conclusion Measurement uncertainty in complete blood cell could be estimated by using IQC data associated with SNCS comparative data , which is in accord with the instrument of target measurement uncertainty .

At present,the doctor-patient relationship in China is a little tense,communication between doctors and patients is not smooth,and the doctor-patient communication skills of the young doctor is obviously insufficient and need to be improved.In this paper,we analyze the factors that affect the communication between doctors and patients in the Department of Neurology through the complexity of professional knowledge,the current background and so on.In addition,we discuss multiple methods of improving communication capability among this group doctors,hoping to strengthen young physicians' clinical basic skills training,and impart doctor-patient communication skills from the linguistic modifiers,oral expression,words and deeds,use of metaphor,simulation training and other aspects,and establish dynamic assessment system of doctor-patient communication ability.

Objective To investigate the effects of vitamin K1 in the adjuvant treatment of spasmodic cough in pertussis syndrome children,and its impacts on length of hospital stay.Methods 87 pertussis syndrome children were randomly divided into two groups,among which 41 cases in the control group were treated with the therapies such as anti -infection,respiratory support,antispasmodic,expectorant et al,while 46 cases in the treatment group were treated additionally with vitamin K1 on the basis of the control group.Then compared the duration and hospital stay of the two groups with spasmodic cough.Results The disappeared time of spasmodic cough in the treatment group was (7.22 ±1.33)d and hospital stay was (9.52 ±1.84)d,which in the control group were (15.51 ±2.73)d,(18.71 ± 3.30)d respectively,and there were statistically significant differences(t =-18.310,-16.269,all P <0.01).In the treatment group,the total effective rate was 93.5%,there were 18 cases with markedly effective,25 cases with effective and 3 cases with ineffective.Correspondingly,there were 7 cases markedly effective,23 cases effective, 11 cases ineffective,and total efficiency rate was only 73.1% in the control group,the difference was statistically significant(χ2 =4.285,P <0.05 ).Conclusion Vitamin K1 to alleviate spasmodic cough in children has a significant effect,which can reduce the duration of the cough time and hospital stay in pertussis syndrome children, and also can improve the clinical effect.

Objective To investigate the relationship between the dynamic changes of platelet parameter and the severity of the neonatal hypoxia ischemic encephalopathy .Methods To search PubMed ,Ovid ,Springer Database ,Chinese Academic Journal ,VIP database and Wanfang database in the library literature and screen documents according to the inclusion criteria of diagnostic tests , then to extract the characteristic information .Using RevMan 5 .1 software to analyze the data ,and to carry out the Meta analysis se-lected the effects model according the results of tests for heterogeneity .Results According to the inclusion criteria ,it was won the 10 articles eventually .The Meta analysis results showed that the PLT in acute period of HIE patients was significantly 1 .65-2 .14 times the standard deviation lower than the control group .The MPV and PDW in acute period of HIE patients were significantly 1 .12-1 .61 and 1 .17-1 .99 times the standard deviation higher than the control group .The PLT in convalescent of HIE patients was 1 .85-2 .35 times the standard deviation higher than the acute period of HIE patients .The MPV and PDW in the convalescent of HIE patients were significantly 0 .68-1 .38 and 0 .81-1 .37 times the standard deviation lower than the acute period of HIE pa-tients .Obviously ,the PLT ,MPV and PDW in the convalescent of HIE patients were closed to the level of the control group .Conclu-sion Clinical observing HIE infant′s platelet parameter dynamically can be regarded as the index of HIE infant′s condition severity and monitoring condition change .

Objective To investigate the clinical symptoms and MRI imaging findings in patients with autoim-mune encephalitis and to improve clinician's understanding about the clinical and imaging characteristics of autoim-mune encephalitis. Methods We analyzed the clinical features and MRI findings of 33 patients with autoimmune en-cephalitis in our department. Results Of these 33 patients, 27 (81.8%) had psychiatric symptoms, 26 (78.8%) had seizure, 18 (54.5%) had involuntary movement, 11 (33.3%) had fever and 9 (27.3%) patients presented with cen-tral hypoventilation, the present of involuntary movement and fever was lower in group LE than in group NMDA. A total of 10 patients had positive MRI finding. Of these 10 patients, 8 had brain parenchyma lesions, 3 had meningeal involvement. The most likely affected parenchymal lesions are occipital lobe, bilateral hippocampus, frontal lobe, pari-etal lobe, temporal lobe, thalamus and cerebellum. Conclusion Psychiatric symptoms and seizure are the most com-mon neurological symptoms of autoimmune encephalitis. MRI may show abnormal signals in the limbic system. FLAIR is the most sensitive MR imaging sequence for detection of autoimmune encephalitis lesions.

Objective To explore the role of LiCl in modulating bacterial-mediated inflammation after Pseudomonas aeruginosa infection.Methods Western-blot was used to determine the efficacy of LiCl usage.The expression of inflammatory cytokines in Pseudomonas aeruginosa-infected macrophages and neutrophils was detected by qPCR.Cell apoptosis was measured by flow cytometry.Results Western-blot data showed that LiCl up-regulated the protein levels of p-GSK-3β(Ser 9)and β-catenin in macrophages and neutrophils,indicating the efficacy of LiCl usage.qPCR data indicated that LiCl enhanced the expression of anti-inflammatory cytokines and suppressed the expression of pro-inflammatory cytokines in Pseudomonas aeruginosa-infected macrophages and neutrophils.Flow cytometry data indicated that LiCl could promoted the apoptosis of Pseudomonas aeruginosa-infected macrophages and neutrophils.Conclusion LiCl inhibited the Pseudomonas aeruginosa-induced inflammation,via regulating the inflammatory cytokine expression and the apoptosis of inflammatory cells.

AIM: To investigate the differentiation from adult rat and human bone marrow mesenchymal stem cells (BMMSCs) into neuron with musk polypeptide (Mu-P).METHODS: Adult rat and human BMMSCs were induced with Mu-P.Neuron-specific enolase (NSE),neurofilament (NF),Nestin,glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry.RESULTS: Simple methods with Mu-P induced adult rat and human BMMSCs exhibiting a neuronal phenotype,expressing Nestin at 3 hours to 5 hours,and expressing NE and NF at 5 hours to 7 days.But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION: Adult rat and human BMMSCs can be induced to differentiate into neurons with Mu-P.

AIM: To investigate multi-potential of rat bo ne marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC w ere long passaged in vitro. METHODS: Cellular cycles of diff erent passages were assayed by FA CSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS: The early passages and long-term passages all showed st rong proliferation; passage 6, passage 25 and passage 45 all showed normal karyo type. CONCLUSION: Long-term culture and passage of rBMMSC still remain s strong proliferation. With this capability, the mutation inclination is not enhanced.

Objective To discuss the influence of special nursing intervention for the quality of life of outpatients with chronic pelvic inflammatory disease. Methods Divided 80 patients into the observation group and the control group randomly, there were 40 cases in each group. Routine nursing, treatment and disease-related lecture were used in both the groups, while the combined therapy of Chinese and Western medicine was used in the observation group in addition. Evaluated the quality of life and the effects of treatment by the World Health Organization′s quality of surviving chart between the two groups after 3-month intertention and 6-month follow-up. Results The scores of quality of environment field, physiology field ,psychological field, sociological field, independence field and mental field in the observation group was (70.33+16.85 ) (70.38+18.80), (71.43+18.28),(70.13+19.08), (73.10+17.70) and (75.38+7.30) respectively, while in the control group was (61.38+16.17), (56.70+17.69), (43.13+19.77), (55.00+19.08), (53.75+16.45) and (49.80+17.26) respectively, the difference between the two groups was significant, the t values were 2.424-8.631, P < 0.05. The effective rate of treatment in the observation group was 95.00%(38/40), while in the control group was 82.5%(33/40), the difference was significant, U=2.507, P < 0.05. Conclusions Special nursing intervention can promote the quality of life ,effects of treatment and the satisfaction of treatment in outpatients with chronic pelvic inflammatory disease, improve their psychological state and the compliance.