Objective The effects of nemo-like kinase(NLK)in inflammation of central nervous system were explored. Methods An animal model with central nervous system inflammation disease induced by intracerebroventricular infusion of lipopolysaccharide was constructed. A model of neuronal apoptosis induced by glutamate in PC12 cells was constructed. A pcdna 3.1-NLK over-expression plasmid and a pSilencer 4. 1-CMV-NLK small interfering plasmid were also constructed. The expression and location of NLK were detected using western blot analysis, double immunofluorescent staining, and cell counting kit-8 assay. Results The protein level of NLK was reduced after induction of inflammation in the brain(t =2. 718-3. 106, all P ＜0. 01), and NLK was only expressed in both cortical and hippocampal neurons. At the cellular level, NLK expression was gradually reduced along with neuronal apoptosis induced by glutamate in PC12(t =4. 032, P ＜0. 01). Over-expression of NLK would reduce the apoptosis of neurons induced by glutamate(t =3. 930, P ＜ 0. 01). Conversely, interfering the expression of NLK would enhance neuronal apoptosis(t = 2. 845, P ＜ 0. 01). Conclusion NLK can protect neurons by inhibiting apoptosis in the process of central nervous system inflammation.

Objective To make suggestions of reform of nursing postgraduates' training mode according to the state of their core competence. Methods 297 nursing postgraduates were investigated using the assessment form of nursing postgraduate's core competence, the questionnaire of training mode and general conditions. Results Nursing postgraduate's core competence were not good, the four lowest scored ability respectively were English ability, scientific research ability, teaching ability, grasping ability of professional knowledge and skill. Conclusions In order to promote the teaching quality, we should reform the training mode from the following aspects: to make clear training target; to make clear professional direction, formulate and carry out training program effectively; to increase the courses related with profession; to reinforce the study of public English and professional English ; to improve the ability of scientific research; to strengthen and perfect the teaching practice; to consolidate the basic knowledge,the basic theory and the basic skill of nursing; to build a unified and normal evaluation system.

Objective To explore the influence of Chinese Qigong--Baduanjin on mild cognitive impairment (MCI) in elderly diabetic patients.Methods A total of 78 elderly type 2 diabetic patients complicated with MCI were divided into intervention group and control group by random digits table method.The control group with 41 patients was received routine treatment and discharged guidance after discharging from hospital.The intervention group with 37 patients was received Baduanjin exercise for 12 months based on the routine treatment in control group.Montreal Cognitive Assessment (MoCA) and Activity of Daily Living (ADL) scores in two groups were compared before intervention and 1,3,6,9 and 12 months after intervention.Results In intervention group,MoCA scores were significantly increased at different times after intervention compared with those before intervention,F=72.782,the differences were statistically significant,P＜0.05.At the same time,MoCA scores were all significantly higher in intervention group than those in control group,F=2.231,2.972,4.362,5.085,5.373,the differences were statistically significant,P＜ 0.05.In intervention group,ADL scores had significantly improved since 6 months after intervention compared with those before intervention,F=93.126,the differences were statistically significant,P＜0.05,and compared with those in control group,the scores were significantly higher since 6 months after intervention,F=3.853,4.561,7.162,the differences were statistically significant,P＜0.05.Conclusion Chinese Qigong--Baduanjin in treatment of elderly diabetic patients complicated with MCI can delay the MCI progress and improve ADL of the patients.

Objective To analyze the specific peptide of cystatin C (CysC) and its characteristics by bioinformatics technology,and verify the predicted results by mass spectrometry.Methods Online software was applied to analyze the physicochemical properties and homology of CysC peptides hydrolyzed by trypsin and predict the associated parameters of ionized fragmentation of specific peptide by mass spectrometry.Precursor ion scan and product ion scan were conducted on the samples of synthetic specific peptide.The recombinant human CysC and serum samples were analyzed by mass spectrometry after trypsin digestion.The results of analysis were compared with the outcomes predicted by bioinformatics.Results T3 (ALDFAVGEYNK) was considered as the specific peptide of CysC by software analysis.When selecting[M + 2H] 2 + for product ion scan,almost all the y and b ions of fragmentation were observed using tandem mass spectrometry (MS/MS),showing consistency with Skyline predictions.Moreover,both the peptides from the human recombinant CysC and serum sample following the trypsin digestion were eluted at the same time with the isotope-labeled T3 * under the fixed conditions.Conclusion Bioinformatics technology could be available for picking out the specific peptides of target protein quickly and efficiently and predicting the ionized fragmentation precisely by mass spectrometry scanning.

OBJECTIVE:To observe clinical efficacy and safety of Kangfuxin liquid in the treatment of pediatric hand,foot and mouth disease. METHODS:128 children with hand,foot and mouth disease were randomly divided into observation group and control group,with 64 cases in each group. Control group was given routine treatment as herpes and oral care,antibiotics for infec-tive children reflected by hemogram elevation,fluid infusion and non-steroidal analgesic-antipyretic agent based on disease condi-tion. Observation group was additionally given Kangfuxin liquid,3 ml for below 3 year-old,tid and 5 ml for more than 3 year-old,tid. Clinical efficacy of 2 groups were observed as well as CRP,lactic acid,Ig,the level of myocardial enzyme before and after treatment;the time of symptoms and signs disappearance and ADR were compared between 2 groups after treatment. RE-SULTS:The total effective rate of observation group(89.06%)was significantly higher than that of control group(70.31%),with statistical significance(P<0.05). There was no statistical significance in CRP,lactic acid,Ig and myocardial enzyme before treat-ment and IgM level after treatment between 2 groups,with statistical significance(P>0.05). CRP,lactic acid creatine kinase and lactate dehydrogenase level of 2 groups decreased significantly,while IgA and IgG levels increased significantly,and the observa-tion group was higher than the control group,with statistical significance(P<0.05). Antipyretic time,hand foot skin rash subsided time and oral ulcer healing time of observation group were significantly shorter than those of control group,with statistical signifi-cance(P<0.05). No obvious ADR was found in 2 groups during treatment. CONCLUSIONS:Kangfuxin liquid is effective in the treatment of hand,foot and mouth disease in children,and can effectively improve CRP,Ig,lactic acid,and myocardial enzyme level with good safety.

Objective To establish the methodology of uncertainty evaluation with reference measurement procedure by Monte Carlo method (MCM).Methods According to JCGM 101:2008,we established the methodology of uncertainty evaluation by MCM in the example of GGT reference measurement procedure.We could calculate an estimate of GGT concentration,the associated standard uncertainty and a coverage interval with a specified probability by MATLAB software,setting the uncertainty evaluation by GUM method as a control.Results When the uncertainty was evaluated by GUM method,the results of sample A and Sample B were (95.8 ±2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L (k =2) with coverage interval [176.1 U/L,183.9 U/L] respectively,while using MCM method,the uncertainty evaluation result of sample A and Sample B were (95.8 ± 2.4) U/L (k =2) with coverage interval [93.4 U/L,98.2 U/L] and (180.0 ± 3.9) U/L with symmetrical 95％ coverage interval [176.2 U/L,183.8 U/L].The output quantity simulated by MCM was normal distributed.Conclusions When the distribution of the output quantity is normal,the measurement uncertainty evaluated by both MCM and GUM method is nearly the same.When the distribution of the output quantity is unknown,MCM can be used as a verification of GUM method.

Objective To observe the month age distribution of peroxisome proliferators activated receptor gamma (PPARγ) expression in rat kedney. Methods Wistar rats aged 3 months,12 months and 24 months were made as models who represented young, middle-aged and old group respectively. Western blotting, immunohistochemical (IHC) and in-situ hybridization (ISH) were used to detect the expression and location of protein and mRNA of PPARγ in rat kidney. Results Western blotting results showed that the expression of PPARγ protein was higher in 3 months group than in 24 months group (0.94±0.05 vs. 0.78±0.02, P＜0.01) and 12 months group (0.87±0.04, P＞0.05), and it reduced in 24 months group than in 12 months group (P＞0.05). By IHC,the PPARγ protein was localized predominantly in the nuclear of tubular epithelia and collecting duct cells in each group. In old age group, PPARγ protein was also detected little in the mesangial and Bowman's capsule epithelial cells. Meanwhile, the distribution of PPARγ mRNA with ISH was consistent with above findings. Additional, semi-quantitative analysis of ISH results verified that the level of PPARγ mRNA decreased with ageing. Conclusions As a nuclear transcription factor,PPARγ participates in the regulation of rat kidney aging.

AIM: To evaluate the effect of carbon monoxide(CO) on the permeability of brain blood barrier(BBB) in cerebral ischemic rats. METHODS: SD rats were divided into three groups. Saline, hemin or ZnPP were injected intraperitoneally 12 h before middle cerebral artery occlusion (MCAO), respectively. The concentration of blood CO and the permeability of BBB at 24 h after MCAO were measured. RESULTS: The CO concentration in blood in hemin group was higher than that in saline group( P 0.05). CONCLUSION: CO reduced the permeability of BBB as a messenger gas molecular when its intrinsic concentration was elevated.

Objective:To evaluate the impact of sample pooling strategy on 2019-nCoV RNA detection results.Methods:Ten negative swabs were stored in 6 ml virus transport medium, mixed thoroughly and diluted 1∶2 and 1∶10. Inactivated 2019-nCoV culture medium was added to simulate pooling samples: 10 pooling samples, 5 pooling samples and 1 swab sample. Extraction and amplification were made using three nucleic acid extraction reagents a, b, and c with different extraction methods and systems, as well as five 2019-nCoV detection reagents A-E with various template loading volumes and sensitivities respectively.Results:For the same sample, the Ct values of extracted templates a were 2.10±0.47 and 3.46±0.62 earlier than extracted templates b and c. For samples with identical amplifying, the Ct valves of N and ORF1ab gene of A reagent were 1.16±0.48 and 2.36±0.54 earlier than that of reagent B. Adding nucleic acid of 10 negative swabs to the amplification system lagged the Ct values of reagent A by about 1.36±0.32 Ct, while Ct values of reagent B were not affected. Extracted by regent a, a lag of 1.66±0.39 Ct on average was observed in C, D, and E reagents in detecting pooling samples of ten swabs as compared with one swab sample. When extracting 400 copies/ml pooling samples of ten swabs by reagent a, N gene could be detected by reagents C and E, but not by reagent D.Conclusion:Large amount of extraneous DNA is introduced by sample pooling, which could interfere the effiency of extraction and amplification. Strategies of using extraction reagents with large loading volume and high effiency, together with amplification reagents with large template volume and low limit of detection are helpful for ensuring detection sensitivity of pooling samples, and greatly reducing the risk of false negative results.

Objective To establish a time-resolved immunofluorometric assay (TRFIA) to detect seurm CⅣ(collagenⅣ). Methods The antibodies to CⅣwere coated on mircoplate and the europium-labeled monoclonal antibody of CⅣ. The luminescent enhancement system was used as enhancement solution which contained mainly 2-naphthoy trifluoroacetone. we established A sandwich time-resolved fluoroimmunoassay (TRFIA) was established to measure the seurm CⅣin 127 patients with hepatitis and 30 normal controls. Results The sensitivity of assay was 12. 8?g/L. The coefficient of variation for inner-batch and inter-batch were 4. 54% and 8. 06%,respectively. The recovery was 98. 6%. The serum level of CⅣwas 46. 06?22. 21?g/L in normal control,47. 25?22. 58?g/L in acute hepatitis, 129.01?53.68?g/L in mild chronic hepatitis,277. 90?92.36?g/L in moderate chronic hepatitis,413.90?162.24?g/L in serious chronic hepatitis,568. 60?210.40?g/L in liver cirrhosis. As compared to normal control,higher concentrations of CIV (P