Construction of a Spink5 conditional knockout mouse model and analysis of its phenotype / 中华皮肤科杂志

Objective:To construct a serine protease inhibitor Kazal type-5 (Spink5) conditional knockout mouse model, and to identify its phenotype.Methods:B cell-specific Spink5 conditional knockout mice of genotype Mb1 cre/+Spink5 floxp/floxp were constructed by using clustered regularly interspaced short palindromic repeats (CRISPR) /CRISPR-associated protein 9 (Cas9) technology, and served as the knockout group. Mice of genotype Mb1 +/+Spink5 floxp/floxp served as the control group. The mice of genotype Mb1 cre/+Spink5 floxp/floxp or Mb1 +/+Spink5 floxp/floxp were sacrificed when they were 4 to 6 weeks old, splenic mononuclear cells were isolated, and B lymphocytes and non-B lymphocytes were sorted by flow cytometry and fluorescence-activated cell sorting. Genotype identification was performed by PCR, and protein expression of lymphoepithelial Kazal-type-related inhibitor (LEKTI) was determined by Western blot analysis. Skin tissues were resected from the mice, and subjected to hematoxylin-eosin staining for measuring the epidermal thickness. Immunofluorescence staining was performed to determine fluorescence intensity of LEKTI protein in the mouse skin tissues. Paired t test or two-independent-sample t test was used for comparisons between groups. Results:Genotype identification results demonstrated that the stable B lymphocyte-specific Spink5 conditional knockout mouse model was successfully constructed. Western blot analysis revealed that the relative protein expression of LEKTI in the B lymphocytes in the knockout group was 0.01 ± 0.02, which was significantly lower than that in the non-B lymphocytes in the knockout group (0.66 ± 0.11, t = 9.99, P < 0.001) , and that in the B lymphocytes in the control group (1.08 ± 0.13, t = 13.78, P < 0.001) . Among 39 mice in the knockout group, 4 presented with dry skin and scattered scaly hypertrophic maculopapules. The epidermal thickness of the lesional skin tissues in the knockout group was 90.42 ± 21.31 μm, significantly higher than that of the non-lesional skin tissues in the knockout group (29.71 ± 3.63 μm, t = 5.05, P = 0.002) and that of normal skin tissues in the control group (12.42 ± 2.21 μm, t = 6.74, P < 0.001) . Immunofluorescence staining showed no significant difference in the fluorescence intensity of LEKTI protein among the lesional skin tissues (46.21 ± 1.21) , non-lesional skin tissues (46.62 ± 2.13) in the knockout group and normal skin tissues in the control group (47.69 ± 1.71, P > 0.05) . Conclusion:The B lymphocyte-specific Spink5 conditional knockout mouse model was successfully constructed, which provides a basis for further exploring mechanisms underlying skin barrier defects and immune dysfunction in Netherton syndrome.

Relationship of clusterin expression with Bax and p53 expression in non-small cell lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>It has been proven that clusterin is a newly apoptosis-related factor and upregulates in many tumors. It plays important roles in carcinogenesis and tumor progression. The antiapoptosis of clusterin seems to be relative to other antiapoptosis factors. The aim of this study is to investigate the expression and significance of clusterin in non-small cell lung cancer (NSCLC) tissue.</p><p><b>METHODS</b>The expression of clusterin, p53 and Bax in NSCLC were detected by immunohistochemical SP method and Western blot assay.</p><p><b>RESULTS</b>Positive expression rate of clusterin was 79.25% (42/53) in NSCLC tissues which was much higher than that in normal lung tissue (2/16, 12.50%) (Chi-square=23.68, P < 0.05). The expression of clusterin was closely related to pathological differentiation (rs=0.464, P < 0.01), clinical stage (rs =0.320, P < 0.01) and lymph node metastasis (rs=0.255, P < 0.05), but not correlated to the sex (Chi-square=0.007, P > 0.05), age (Chi-square=0.707, P > 0.05) and histological type (Chi-square=0.702, P > 0.05). The expression of clusterin in NSCLC was positively correlated to the expression of p53 (rs=0.589, P < 0.01), but was negatively related to the expression of Bax (rs =-0.346, P < 0.01). The relative expression level of clusterin protein in NSCLC was significantly higher than that in normal lung tissue (0.541±0.010 vs 0.201±0.020) (P < 0.05).</p><p><b>CONCLUSIONS</b>Clusterin may play an important role in the biological characteristics of NSCLC by the antiapoptosis pathway.</p>

Social function cognition for bedside care service at patient's home in the residents aged over 55 years in Jingshan Community of Bering / 中华全科医师杂志

Objective An investigation on social function cognition for bedside care service at patient's home was conducted in the residents aged over SS years in Jingshan Community,Dongcheng District,Beijing,to lay a scientific basis for provision of it.Methods Ninety-seven residents aged over 55 years were selected from 58 families with systematic sampling in Jingshan Community by a questionnaire interview,including history of illness,income,medical expense,awareness of bedside service at patient's home,etc.Results Totally,98.9 % of the residents suffered from at least one kind of chronic disease,98.9 % of them never received bedside care service at their homes,94.9% of them only knew a little of such kind of health care service,91.85% of them knew some social function of it,and 84.5 % of them did not know the criteria of payment for the service.Conclusions Publicity of bedside care service at patient' s home should be strengthened,focusing on its social function,inexpensiveness,convenience,etc.Function of bedside service should be perfected further,with quality care,to make it an important part of community health care.