OBJECTIVE:To provide references for the legislation of Licensed Pharmacist Legislation. METHODS:Several misunderstandings appearing during the complementation of licensed pharmacist regulation were analyzed, as well as the reasons for these misunderstandings. RESULTS&CONCLUSION: The misunderstandings in the respect of legislative subjects, licensed posts, licensed treatment, talent market demand and further education should be avoided in the process of Licensed Pharmacist Legislation.

Objective Inquire into the relationship between diabetic peripheral neuropathy(DPN)pathogenic factor and Antigangliosides antibody(Anti-GS-Ab)in type 2 diabetes mellitus. Methods It was examined by enzyme-linked immunosorbent assays(ELISA)to the levels of serum Anti-gangliosides(Anti-GS)in 2 DM and DPN as well as healthy.Results The positive rate of Anti-GS-IgM,IgG in DPN group were 46.7% and 20.0%.repectivily it was obviously higher than normal group and 2 DM group.Conclusion The relationship between the DPN and Anti-GS-Ab is a close.It show that Anti-GS-Ab play an important role in DPN pathological process.

BACKGROUND:Bone marrow mesenchymal stem cel s have a low survival rate after implanted into the ischemic myocardium. However, hypoxia preconditioning (HPC) may enhance bone marrow mesenchymal stem cel proliferation and promote its survival rate. OBJECTIVE:To explore whether Pim-1 is involved in HPC protecting against apoptosis of bone marrow mesenchymal stem cel s and the relevant mechanism. METHODS:Bone marrow mesenchymal stem cel s were respectively subjected to HPC for 0, 6, 12, and 24 hours. The expression of Pim-1 and apoptosis-related genes were detected by RT-qPCR and western blot. Then, the best hypoxic preconditioning time was determined as 12 hours. Then, bone marrow mesenchymal stem cel s were assigned to one of the fol owing groups:control (without HPC), 12-hour HPC, 12-hour HPC+Pim-1 inhibitor groups. Flow cytometry analysis was used to detect the cel apoptosis, Transwel assay to analyze the cel migration ability in each group, and JC-1 kit to detect mitochondrial membrane potential. Animal models of myocardial infarction were established. One week after modeling, bone marrow mesenchymal stem cel s were given via multi-point injection around the infarct zone of rats. Two weeks after modeling, heart tissues of rats were taken and sliced fol owed by DiI staining to calculate the survival rate of bone marrow mesenchymal stem cel s. Additional y, rat cardiac function was assessed by echocardiography prior to and after modeling as wel as at 4 weeks after cel transplantation. RESULTS AND CONCLUSION:At 12 hours after HPC, the expression of Pim-1, p-Akt and Bcl-2 gene in the infarct region was significantly increased, but the expression of caspase-3 and Bax was significantly decreased. Compared with the control group, cel viability in the 12-hour HPC group was increased very significantly at 1 week after cel transplantation (P<0.001), the migration and anti-apoptosis ability were enhanced significantly (P<0.01) and the cardiac function of rats was significantly improved in the 12-hour HPC group (P<0.05). Al of these protective effects were blocked by the Pim-1 inhibitor. These findings indicate that HPC can protect bone marrow mesenchymal stem cel s from apoptosis through activating Akt and up-regulating Pim-1, and thereby improve the therapeutic effect of bone marrow mesenchymal stem cel transplantation on ischemic heart diseases.

Objective To study the role of peripheral blood T regulatory cells (Treg) markers, as well as different Treg cells in the pathogenesis of systemic lupus erythematosus (SLE) and explore the correla-tion betweenCD127 and Foxp3. The immunosuppressive effect of CD4~+CD25~+CD127~(low/-) T cell is explored. Methods ①Four-color direct fluorescence-labeled and multi-parameter flow cytometry were used to detect peripheral blood CD4~+CD25~+ T cells and other Treg cells accounted for the proportion of CD4~+ T cells in 40 SLE patients (19 cases with active disease 21 cases in remission) and 15 healthy controls. Meanwhile, its correlations with anti-dsDNA antibodies among 7 groups were analyzed.②Flow cytometry sorting combined with cell culture were applied to detect and analyze the proliferation inhibition effect of CD4~+CD25~+CD127~(low/-)regulatory T cells to CD4~+CD25~-effector T cell.Two independent samples t test,ANOVA for repeated measures,Pearson's correlation and Spearman's correlation were used for statistical analysis.Results ①The cell ratio of the 7 groups of SLE patients was(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%(0.97±O.28)%,(0.69±0.23)%and(O.71±0.35)%respectively.In the SLE group,the proportion of the first 6 groups was lower than the control group (P<0.05).For CD4~+CD127~(low/-)Foxp3~+,there wag no difference in the two groups(P>O.05).②CD4~+CD25~+Foxp3~+,CD4~+CD25~(high)Foxp3~+ T cells and IgA ratio Was positivelv correlated.While CD4~+CD25~(high)CD127~(low/-) T cells and anti-SSB antibodies was positively correlated in SLE patients.③The seven group of cells,in addition to CD4~+CDl27~(low/-)Foxp3~+ T cell ratio were lower in early-onset SLE patients than those in patients at remission(P<0.05).④The cell propor-tion of the first 6 groups was lower than that of post steroid treatment(P<0.05).⑤Foxp3 expression of CD4~+CD25~+T ceils and CD4~+CD25~(high) T cells was positively correlated with low expression of CDl27 in the early onset group,remission group and the control group.⑥The CD4~+CD25~- effect T cells could be suppressed by their own CD4~+CD25~+CDl27~(low/-) regulatonry T cells in vitro,and the inhibition of SLE patients was significantly lower than controls.Conclusion The immunological abnormality of SLE may be associated with the qnatity and functional defects of immune regulatory T cells.CDl27 may be a possible alternative to Foxp3 regulatory T cell-specific surface markers.

Objective To study the pathological alterations,such as oxidative stress,cell proliferation and insulin resistance,especially autophagy,in Alzheimer' s disease (AD) complicated with type 2 diabetes (AD + T2DM).Methods The mouse models of T2DM,AD and AD + T2DM were used in the study,and totally 80 mice were divided into four groups:control group,T2DM group,AD group and AD + T2DM group.Morris water maze was applied to test the ability of learning and memory among the above mentioned groups.In the meantime,insulin resistance index,the expression of insulin receptor substrate 2,oxidative stress,cell proliferation and autophagy were observed with chemical analysis,immunofluorescent labeling,transmission electron microscopy and Western blotting.Results On day 4,the difference of time to find Morris water maze in control group,T2DM group,AD group and AD + T2DM group ((26.08 ±4.93) s,(38.46 ± 4.07) s,(47.32 ± 5.86) s,(53.01 ± 6.12) s,F =2.454,P =0.025) was statistically significant,and the time in AD + T2DM group was longer than that in AD group (t =-3.624,P =0.033).Compared with control group,insulin resistance occurred in T2DM group,AD group and AD + T2DM group (4.35 ± 0.48,16.12 ± 3.57,7.03 ± 3.11,18.78 ± 5.06,F =5.602,P =0.009),and the reduction of insulin receptor substrate 2 expression,the oxidative stress reaction,neural proliferative suppression and autophagy (F =418.344,222.514,436.250,113.934,23.772,35.469,all P ＜ 0.05) were induced in T2DM group,AD group and AD + T2DM group,which were more serious in AD + T2DM group than in AD group (t =-2.796,21.723,-8.041,9.037,-4.403,-32.011,-26.593,all P ＜0.05).Conclusion AD + T2DM mice suffered more serious cognitive impairment than AD and T2DM mice.The oxidative stress levels of AD + T2DM mice were upregulated,and thus led to the inhibition of cell proliferation,eventually leading to promotion of autophagy.

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

Objective To evaluate the application of Web problem-based learning (WPBL) mode of respiratory system integration curriculum.Methods Experimentclasses 1 and 2 of the 2014 grades of the Qiqihar Medical University were divided into control group(48 students) and experiment group (48 students).Traditional PBL teaching was conducted in control group:the contents of the case were printed and discussed with the material of each class.WPBL teaching was conducted in experiment group:autonomous learning before class,showing cases by Video,asking questions and discussion in group and reporting results in class.The teaching effect was evaluated by PBL evaluation form,questionnaire and final grade.SSPS 19.0 statistical software was used for data analysis and measurement.T test was conducted among groups.Counting data were tested by Chi-square and the results were expressed as percentage.Results The students in the experiment group were better than those of control groupin the aspects of contents understanding,course interest,clinical thinking ability,classroom atmosphere and the course attitude.The scores of PBL (16.65 ±2.82),experiment exam (8.21 ±0.44) and final grade(76.77 ± 12.31) in the experiment group were higher than those in the control group.There are statistical differences in two groups (P＜0.05).Conclusions WPBL teaching can improve students'learning interest,clinical thinking ability and improve the teaching quality of respiratory integration curriculum.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.