Expression features of cytochrome C oxidase subunit Ⅰ, BCL2 interacting protein 3 and interleukin-1β in MRL/lpr lupus mice liver tissue / 中华风湿病学杂志

Objective:To explore the expression features of cytochrome C oxidase subunit Ⅰ (MT-CO1), BCL2 interacting protein 3 (BNIP3) and interleukin (IL)-1β in the liver of MRL/lpr lupus mice.Methods:The mRNA and protein levels of MT-CO1, BNIP3, IL-1β, p16 and p21 in lupus mice and control mice were detected by polymerase chain reaction (PCR) and Western blot, the IL-1β expression site were detected by hematoxylin and eosin (HE) staining and immunohistochemical method, and themalondialdehyde (MDA) was detected by colorimetry. Hepatocytes and macrophages were stimulated with lipopolysaccharide (LPS), while hepatocytes were also cultured with supernatants obtained after macrophages stimulated with LPS, and the mRNA and protein levels of MT-CO1, BNIP3 and LC3B, as well as p16 and p21 expression, were determined by qPCR and Western blot. The expression of mitochondrial reactive oxygen species (mtROS) was detected by immunofluorescence. One way Analysis of Variance (ANOVA) was used to compare the mean of each group, and LSD method was used to compare the means of multiple samples, and Tamhane's T2 method was used to compare the means of multiple samples when the variance was uniform. Results:The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the liver tissue of the lupus group (0.14±0.04; 0.16±0.05) were significantly lower than those of the control group (0.11±0.04; 0.16±0.06), and the differences were statistically significant ( t=7.16, P<0.001; t=4.54, P<0.001). The expression levels of IL-1β, p16 and p21 in the lupus group (2.06±0.69; 0.37±0.14; 0.16±0.06) were significantly higher than those of the control group (0.23±0.06; 0.25±0.08; 0.11±0.04) ( t=9.58, P<0.001; t=24.35, P<0.001; t=22.36, P<0.001). The results of Western blot were consistent with those of PCR. HE staining showed lymphocyte infiltration in the liver tissue of lupus mice, and immunohistochemistry showed IL-1β in the liver tissue of lupus mice. The positive cells were mainly concentrated in the sinusoids, and the expression of hepatic parenchymal cells was not rearkable. The content of MDA in liver tissue of the lupus group (0.19±0.10) was higher than that of the control group (0.17±0.09), and the difference was statistically significant ( t=4.33, P=0.005). LPS directly stimulated AML12 hepatocytes (0.069±0.028; 0.17±0.07). The PCR results showed that compared with the control group (0.176±0.072; 0.08±0.03), the expression of MT-CO1, and BNIP3 were not significantly different ( t=1.01, P=0.337; t=0.88, P=0.399). The expression of IL-1β was significantly higher when incubated with the supernatants of LPS stimulated macrophages (0.28±0.09) compared than that of the control group (0.15±0.05) ( t=28.26, P<0.001). The results of PCR showed that the mRNA levels of MT-CO1 and BNIP3 in the LPS stimulated group (0.046±0.026; 0.17±0.05) were significantly lower than those in the control group (0.143±0.083; 0.18±0.06), and the differences were statistically significant ( t=7.52, P<0.001; t=4.24, P<0.001), The expression of p16 and p21 in LPS stimulated group (0.29±0.09; 0.27±0.09) were significantly higher than those in the control group (0.18±0.06; 0.22±0.07) ( t=13.54, P<0.001; t=8.69, P<0.001). The results of Western blot were consistent with those of PCR. Immunofluorescence showed that the fluorescence intensity of mtROS in LPS stimulated group (0.25±0.10) was higher than that in the control group (0.08±0.03), and the difference was statistically significant ( t= 4.86, P<0.001). Conclusion:Immune-mediated inflammation in the liver tissue of lupus mice can stimulate liver parenchymal cells to cause intracellular mitochondrial dysfunction. However, the mechanism of liver organ damage in lupus mice is not limited to the immune-mediated inflammation of immune active cells, but also include parenchymal cell mitochondrial dysfunction.

Application of imaging techniques in diagnosis and severity assessment of dermatomyositis / 中华皮肤科杂志

Dermatomyositis is an autoimmune disease involving the skin and muscles. At the onset of dermatomyositis, it is difficult to make an early diagnosis due to atypical clinical manifestations and lack of serological markers. Skin and muscle lesions are associated with disease activity and prognosis in patients with dermatomyositis or clinical amyopathic dermatomyositis. Computed tomography, magnetic resonance imaging, ultrasonography, dermoscopy and other imaging techniques may be used to assess skin and muscle involvements, which can not only improve the accuracy of early diagnosis of dermatomyositis, but also provide important reference for the assessment of disease activity and prognosis.

Local infiltration of tissue-resident memory CD4 + T cells in lesions of patients with pemphigus and its clinical implications / 中华皮肤科杂志

Objective:To investigate the local infiltration of tissue-resident memory CD4 + T (CD4 + T RM) cells in lesions of patients with pemphigus and its clinical implications. Methods:From September 2017 to December 2018, 20 patients with pemphigus and 15 healthy human controls were collected from Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. Flow cytometry was performed to determine the proportion of CD4 + T RM cells in skin lesions of pemphigus patients and normal skin of healthy controls. The degree of CD4 + T RM cell infiltration in skin lesions was compared among different body sites of the patients with pemphigus, and the correlation between the proportion of CD4 + T RM cells and the time to disease control was analyzed. Normally distributed data were analyzed by using t test, and non-normally distributed data by using non-parametric test; the Pearson correlation coefficient was used to analyze correlations of the proportion of CD4 + T RM cells with pemphigus disease area index (PDAI) scores and circulating anti-desmoglein (Dsg) antibody titers. Results:Among the 20 patients, there were 16 with pemphigus vulgaris and 4 with pemphigus foliaceus. All the patients had skin involvement, 14 lesional tissue samples were taken from the trunk, and 6 from the limbs. There was no significant difference between the healthy control group and pemphigus group in terms of age, gender or biopsy sites (all P > 0.05) . The proportions of CD3 + T cells (72.75% ± 8.22%) and CD4 + T RM cells (44.05% ± 14.27%) in the skin lesions of patients with pemphigus were significantly higher than those in the skin tissues of the healthy controls (31.33% ± 8.72%, 12.60% ± 5.12%, t = 14.24, 9.10, respectively, both P < 0.001) . Among the patients with pemphigus, the proportion of CD4 + T RM cells was significantly higher in the skin lesions on the trunk (49.57% ± 12.32%) than in those on the limbs (31.17% ± 9.75%, t = 3.23, P < 0.05) . The proportion of CD4 + T RM cells in the skin lesions was positively correlated with the PDAI scores ( r2 = 0.246, P = 0.026) , but not correlated with serum titers of circulating anti-Dsg1 ( r2 = 0.137, P > 0.05) or anti-Dsg3 ( r2 = 0.162, P > 0.05) antibodies in the patients. During the treatment with systemic glucocorticoids, the proportion of CD4 + T RM cells in the skin lesions was significantly higher in the patients whose lesions could not be controlled within 4 weeks than in those whose lesions could be controlled within 4 weeks ( t = 3.22, P < 0.05) . Conclusion:The proportion of CD4 + T RM cells markedly increased in the skin lesions of patients with pemphigus, which may be related to the severity of the disease and response to treatment.

Clinical features of anti-signal recognition particle antibody-positive patients with dermatomyositis or clinically amyopathic dermatomyositis / 中华皮肤科杂志

Objective@#To investigate the clinical features of anti-signal recognition particle (SRP) antibody-positive patients with dermatomyositis/clinically amyopathic dermatomyositis (DM/CADM) .@*Methods@#Clinical data were collected from 90 patients with DM/CADM, who were hospitalized at the Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from June 2015 to July 2017. Immunoblotting assay was performed to determine the serum level of anti-SRP antibody in these patients. Statistical analysis was carried out using t test and Chi-square test.@*Results@#Of the 90 patients with DM/CADM, 11 (12.2%) were positive for serum anti-SRP antibody, including 6 with DM and 5 with CADM. Among 82 adult patients with DM/CADM, the prevalence of malignant tumors was significantly higher in the patients with anti-SRP antibody than in those without (7/9 vs. 31.5%[23/73], χ2 = 7.394, P = 0.006) . The 11 patients with anti-SRP antibody had typical DM skin lesions, and their cutaneous dermatomyositis disease area and severity index (CDASI) was 18.1 ± 2.9. The prevalence of "angel wings sign" (aliform erythema on the trunk) was significantly higher in the patients with anti-SRP antibody than in those without (7/11 vs. 29.9%[20/67], Fisher′s exact test, P = 0.028) . The positive rate of antinuclear antibody was significantly higher in the patients with anti-SRP antibody than in those without (4/8 vs. 16.7%[13/78], χ2 = 6.053, P = 0.014) . Magnetic resonance imaging of muscles of both thighs of the 10 patients with anti-SRP antibody (6 with DM and 4 with CADM) showed the presence of abnormal signals in the thigh muscle group in 8, swelling of the muscle group in 2, subcutaneous edema in 2, myofascial swelling in 1, and no abnormities in 2. No interstitial lung disease or myocardial involvement was observed in the patients with anti-SRP antibody.@*Conclusions@#The anti-SRP antibody-positive patients with DM/CADM showed a high prevalence of "angel wings sign" , and a high risk of malignant tumors. Early detection of the anti-SRP antibody in patients with DM/CADM is helpful to predict the occurrence of malignant tumors.

Pathological features of ectopic lymphoid structure in skin lesions of patients with pemphigus / 中华皮肤科杂志

Objective To investigate pathological features of infiltrating lymphocytes in skin lesions of patients with pemphigus,and to analyze their correlation with titers of anti-desmoglein (Dsg) 1 and anti-Dsg3 antibodies in peripheral blood.Methods A retrospective pathological analysis was performed in 93 patients with pemphigus vulgaris or pemphigus foliaceus,who visited the Department of Dermatology of Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine between 2014 and 2016.For each HE-stained section,the total number of lymphocytes per × 50 microscopic field was counted,and defined as lymphocyte density index.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in the patients with pemphigus.The correlations between the lymphocyte density index and titers of anti-Dsg1 and anti-Dsg3 antibodies were analyzed.Immunohistochemical staining was performed in lesional skin samples from 8 patients with pemphigus vulgaris and 8 patients with pemphigus foliaceus,so as to analyze the distribution of CD3+ T cells,CD20+ B cells and CD138+ plasma cells.Results Of the 93 pathological sections,93 (100.00％) showed Grade1 lymphocyte aggregates,64 (68.09％) showed Grade 2 lymphocyte aggregates,and 10 (10.64％) showed Grade 3 lymphocyte aggregates,and the 56 cases of pemphigus vulgaris and 37 of pemphigus foliaceus showed the similar proportion of grade 1,2 and 3 lymphocyte aggregates.There was also no significant difference in the lymphocyte density index between patients with pemphigus vulgaris and pemphigus foliaceus (P ＞ 0.05),and the lymphocyte density index was uncorrelated with the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in patients with pemphigus.Of the 16 cases of pemphigus,CD3+ T cells were found in all cases,CD20+ B cells in 15,and CD138+ plasma cells in 12.Of 16 sections,all showed a large amount of CD3+ T cells in Grade 1-3 lymphocyte aggregates,while lymphocyte aggregates containing CD20+ B cells and CD138+ plasma cells were found in 52.80％ ± 5.78％ and 34.59％ ± 7.42％ of sections respectively.No significant differences in the distribution of CD3+ T cells,CD20+ B cells,CD138+ plasma cells were found between the 8 cases of pemphigus vulgaris and 8 cases of pemphigus foliaceus.Conclusion Different degrees of lymphocyte infiltration generally exist in skin lesions of patients with pemphigus,which may form ectopic lymphoid structures and contribute to the development and aggravation of pemphigus skin lesions.

Effects of small RNA interference targeting mammalian target of rapamycin on paraquat-induced pulmonary fibrosis in rats / 中华危重病急救医学

Objective To investigate the effects of small RNA interference targeting mammalian target of rapamycin (mTOR) expression on paraquat-induced pulmonary fibrosis in rats.Methods Human embryonic kidney cells HEK-293 were culturedin vitro. The mTOR small interfering RNA (mTOR-siRNA) expression plasmid transfection lentivirus was constructed, and non-specific sequence plasmid with no homology to mTOR gene was set as the control. Seventy-two healthy male Sprague-Dawley (SD) rats were randomly divided into normal saline (NS) control group, paraquatmodel group, mTOR unrelated sequence group, and mTOR-siRNA group, with 18 rats in each group. Paraquat poisoning animal model was reproduced by intraperitoneally injecting 20% paraquat solution 15 mg/kg, while the NS control group was intraperitoneally injected the same volumes of NS. Rats in the mTOR unrelated sequence group and mTOR-siRNA group were injected 1×109 TU/mL lentivirus solution 50μL into the airway, respectively, while in the NS control group and paraquat model group were injected the same volumes of NS. At 7, 14 and 28 days after treatment, 6 rats in each group were sacrificed respectively for lung tissue, the pathological changes and fibrosis of lung tissues were observed under light microscope. The levels of hydroxyproline (HYP) in lung tissues were determined by alkaline hydrolysis. The mRNA and protein expressions of mTOR in lung tissues were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results Under light microscope, there was no obvious pathological changes in the lung tissues in the NS control group, while in the paraquat model group and mTOR unrelated sequence group, lung tissue in rats were damaged, there were a lot of inflammatory cell infiltration, a large number of matrix collagen and fibrous tissues hyperplasia, and gradually increased with time, and it was consistent with paraquat-induced lung tissue fibrosis process. The pathological and fibrotic changes in lung tissue of mTOR-siRNA group were obviously reduced after silencing mTOR gene. The levels of HYP and the expression levels of mTOR mRNA and mTOR protein of lung tissues in the paraquat model group and mTOR unrelated sequence group were continuously increased in time-dependent manner, and they were significantly higher than those in the NS control group at all of the time points, but no significant difference was found between mTOR unrelated sequence group and paraquat model group. In mTOR-siRNA group, silencing mTOR gene could inhibit paraquat poisoning induced HYP increase in lung tissue, and the expressions increase in mTOR mRNAand mTOR protein, the values were close to the levels of NS control group, and the significant difference was found as compared with paraquat model group at 7 days or 14 days, and the change was maintained to 28 days [7 days: HYP (μg/mg) was 1.13±0.06 vs. 1.25±0.07; 14 days: HYP (μg/mg) was 1.19±0.09 vs. 1.29±0.12, mTOR mRNA (2-ΔΔCt) was 0.99±0.11 vs. 1.94±0.12, mTOR protein (gray value) was 0.39±0.08 vs. 0.75±0.09; 28 days: HYP (μg/mg) was 1.28±0.06 vs. 1.40±0.05, mTOR mRNA (2-ΔΔCt) was 1.15±0.13 vs. 2.85±0.15, mTOR protein (gray value) was 0.45±0.10 vs. 0.86±0.12, allP < 0.05].Conclusion Lentivirus-mediated mTOR-siRNA could effectively inhibit the expressions of mTOR in lung tissues of paraquat-poisoned rats, and reduce the damage and fibrosis of lung tissues caused by paraquat.

Application of composite microporous polysaccharide powder in therapy of non-varicose veins of gastroin-testinal bleeding / 实用医学杂志

Objective To evaluate the safety and efficiency of composite microporous polysaccharide pow-der in non-varicose veins of gastrointestinal bleeding treatment. Methods We retrospectively analyzed 35 cases of the past 2 years in this hospital. Those patients were clinical diagnosed with non-varicose veins of gastrointestinal bleeding and received composite microporous polysaccharide powder in hemostasis. Results All 35 patients stopped bleeding after treatment with combined treatment of composite microporous polysaccharide powder spray. All vital signs were smooth and steady such as oxyhemoglobin saturation and heart rate,without complications like irritability,deterioration of inflammation and delayed hemorrhage. Conclusion The combined utilization of com-posite microporous polysaccharide powder provided rapid and effective hemostasis in therapy of non-varicose veins of gastrointestinal bleeding,which is an effective,simple and safe operation and to be worth of being generalized.

Surveillance of schistosomiasis in Sanzhou Village, Dantu District, Zhenjiang City, 2008 / 中国血吸虫病防治杂志

The surveillance of schistosomiasis in Sanzhou Village, Dantu District, Zhenjiang City, a national schistosomiasis surveillance site, showed that in 2008, the area with snails was 27 hm~2, among which the area with infected snails was 14 hm~2, the densities of living snails and infected snails were 0. 86 and 0.002 1 snails/0. 1 m~2, respectively. The infection rate of snails was 0.25% , the positive rate of IHA was 3. 10% and the infection rate of schistosome in human populations was 1%. There were no infected domestic animals found, and there were no acute schistosomiasis and newly advanced schistosomiasis cases found in the surveillance site in 2008. It is indicated that the endemic situation of schistosomiasis is stable. The comprehensive control, including molluciciding and environmental modification, should be implemented for snail control. The comprehensive measures with emphasis on infectious source control should be further strengthened.

Role of Cx43 gene in the process of myocardialization of proximal outflow tract septum in the mouse heart / 中国病理生理杂志

AIM: To investigate the role of Cx43 in the myocardialization of the proximal outflow tract(OFT) septum in the mouse heart.METHODS: C57/BL6 mice of ED11.5 to 1 day after birth were used in this study,which included Cx43 knockout homozygotes((Cx43-/-)),heterozygotes((Cx43+/-)) and wildtypes((Cx43+/+)).Pathohistological analysis was used to examine the structure of the hearts.The expression of alpha-sarcomeric actin(?-SCA),active caspase-3 and activator protein-2(AP-2) were detected by immunohistochemistry.RESULTS: Most(Cx43-/-) mice died within 24 h after birth with a swelling and blockage of the conotruncal region,which led to the obstruction of OFT and enlargement of right ventricle.HE staining showed plenty of abnormal tissues in this region forming many pouches.No apparent malformations were observed in(Cx43+/-) and(Cx43+/+) mice.The expression of ?-SCA in the proximal OFT septum was delayed obviously in(Cx43-/-).The apoptotic cells existed in the proximal OFT septum of(Cx43+/+) mostly during ED12.5 to ED15.5.However,there were less apoptotic cells observed in(Cx43+/-),and few in(Cx43-/-).The expression of AP-2,marker of neural crest cells,was increased in (Cx43-/-) and abnormally located in the proximal OFT septum.CONCLUSIONS: Cx43 KO mice are characterized by hyperplasia in conotruncal region,which may be associated with the delayed myocardialization of OFT septum.The decreased apoptosis and the abnormal distribution of cardiac neural crest cells are likely to contribute to the abnormal myocardialization in mice with Cx43 defects.

Expression of ANGPTL3 mRNA in kidney of adriamycin-induced nephrotic rats / 中华肾脏病杂志

Objective To study the expression of angiopoietin-like protein 3(ANGPTL3) mRNA successively in kidney of adriamycin (ADR)-induced nephrotic rats during the development of proteinuria, and to disclose the possible assosiation of ANGPTL3 with proteinuria. Methods Adriamycininduced nephrotic rat models were established by a single injection of adriamycin via the tail vein. Glomeruli and cortex tubuli of rats were dissected by laser microdissection. Real time quantitative RT-PCR was used to study the expression of ANGPTL3mRNA in kidney of ADR rate at 7, 14, 21 and 28 days successively following adriamycin injection. Results (1) In ADR rats, urinary protein and the level of triglyceride and cholesterol in serum increased significantly at day 14 (P

THE ROLE OF TGF?2 IN MYOCARDIALIZATION OF CARDIAC PROXIMAL OUTFLOW TRACT SEPTUM IN MICE / 解剖学报

Objective To investigate the expression of TGF?2 in proximal OFT septum in connexin(Cx) 43 knockout(KO) mice and illustrate its relationship with myocardialization and reveal whether exogenous TGF?2 could promote myocardialization in vitro.Methods Objects were C57/BL6 mice of E12.5 to E15.5 by the mating of 2 month old Cx43 heterozygous mice,which included Cx43 homozygotes(Cx43-/-),heterozygotes(Cx43+/-) and wild-types(Cx43+/+) genotyped by PCR method.?-sarcomeric actin(?-SCA) and TGF?2 were detected by immunohistochemistry.TGF?2 with 3 different dosages was used to be the intervent added to the heart culture system in which the wild-type hearts of E12.5 were cultured till E15.5 before detecting the expression of ?-SCA by immunohistochemistry.Results The expression of ?-SCA in the proximal OFT septum was delayed obviously in Cx43-/-predominantly at E13.5 and E14.5.As compared with the Cx43+/+,the expression of TGF?2 in proximal OFT septum decreased obviously in Cx43-/-mice predominantly at E14.5,and less obviously in(Cx43+/-) mice.None of the intervention groups showed the obvious promoted myocardialization.Conclusions Cx43 KO mice exhibited delayed myocardialization,and the decreased TGF?2 may involve in the abnormal myocardialization of Cx43 KO mice.Application of TGF?2 in vitro could not exactly mimic the in vivo expression pattern of TGF?2,or the process of myocardialization may require precise dosage of TGF?2 and stringency of cultured system.