Objective To evaluate the effects of tacrolimus on the secretion of IL-6 and sIL-2R as well as the expression of IL-6 and sIL-2R mRNA by lymphocytes.Methods Jurkat human lymphoma cells were cultured and treated with tacrolimus of different concentrations.Enzyme linked immunosorbent assay was performed to determine the levels of IL-6 and sIL-2R in the supernatant of Jurkat cells at 48 hours after treatment with tacrolimus of 0,10,102,103 and 104 nmol/L,and real time reverse transcription PCR to measure the expression of IL-6 mRNA and sIL-2R mRNA of Jurkat cells at 48 hours after treatment with tacrolimus of 102 nmol/L.Results Tacrolimus of 102 - 104 nmol/L could suppress the secretion of IL-6 and sIL-2R from Jurkat cells (all P＜ 0.05),with a more marked suppressing effect achieved by the use of tacrolimus at 103 - 104 nmol/L.The expressions of IL-6 and sIL-2R mRNA from Jurkat cells were downregulated by tacrolimus of 102 nmol/L (both P ＜ 0.05).Conclusion Tacrolimus at certain concentrations could downregulate the secretion of IL-6 and sIL-2R as well as the expression of IL-6 and sIL-2R mRNA by lymphocytes.

Objective To evaluate the role of C-type lectin domain family 2,member B (CLEC2B) gene in the pathogenesis of vitiligo.Methods Real time fluorescence-based PCR was performed to detect the expression of CLEC2B mRNA in the peripheral blood and lesional skin of 37 patients with vitiligo as well as in the peripheral blood and normal skin of 40 healthy controls.Data were statistically analyzed by t test and chisquare test.Results Among the 37 patients,23 had progressive vitiligo,14 stable vitiligo,31 vitiligo vulgaris,6 segmental vitiligo.The expression level of CLEC2B mRNA was significantly higher in vitiligo lesions than in the control skin (1.21 ± 0.03 vs.1.00,t =4.432,P ＜ 0.05),but was of no significant difference in peripheral blood between the patients and healthy controls (1.02 ± 0.05 vs.1.00,t =1.435,P ＞ 0.05).Increased expression of CLEC2B mRNA was noted in lesions of vulgaris vitiligo compared with those of segmental vitiligo (1.21 ± 0.03 vs.1.02 ± 0.01,t =5.330,P ＜ 0.05),as well as in lesions of progressive vitiligo compared with those of stable vitiligo (1.25 ± 0.05 vs.1.08 ± 0.03,t =3.046,P ＜ 0.05).No significant difference was observed in the expression of CLEC2B mRNA among lesions of vitiligo with different courses (P ＞ 0.05).Conclusion The differential expression of CLEC2B mRNA may take part in the pathogenesis of vitiligo.

An 18-year-old female presented with painful erythema and nodules on both legs for more than 2 years.Dermatological examination showed irregularly sized,mildly indurated,tender,deep subcutaneous nodules arising in diffused infiltrated dark erythematous patches in the inner and posterior region of the left leg.Histopathology showed no significant changes in the epidermis.There were perivascular lymphoid cell infiltrates in the dermis and subcutis.Multiple sites of necrosis of blood vessel walls with vascular occlusions were noted.The lumens of some blood vessels were filled with lymphocytes,among which were many atypical cells with hyperchromatic nuclei and pathologic mitotic figures.Immunohistochemistry showed that lymphocytes in the cavities of blood vessels were positive for CD3(+++),CD3ε(+++),CD2(+),CD56(+++),granzyme B(+++),perforin(+++),CD30(+),Ki67 (+++,100％ ),but negative for CD20,CD5,CD7,CD4,CD8,TdT,anaplastic lymphoma kinase,early membrane antigen (EMA) or pan cytokeratin (pCK).The endothelial cells lining the blood vessels stained positively for CD34.The intravascular lymphocytes were also positive for EBER1/2 by in situ hybridization.A diagnosis of cutaneous intravascular NK/T cell lymphoma was made.

Objective:To investigate the clinicopathological characteristics and possible pathogenesis of immune checkpoint inhibitor (ICI) induced myocarditis, and to improve understanding of this new type of myocarditis.Methods:Two cases of ICI induced myocarditis with endomyocardial biopsy available for review were selected from the cases with immune-related adverse events treated by ICI in Peking University First Hospital, Beijing, China from 2020 to 2022. The clinical data, histomorphological characteristics, PD-L1 expression of cardiomyocytes, and classification of inflammatory cells in two cases of ICI-induced myocarditis were analyzed. Relevant literature was reviewed.Results:Case 1 was a 64-year-old male diagnosed with gastric signet ring cell carcinoma. Case 2 was a 56-year-old male ad diagnosed with lung squamous cell carcinoma. Both patients developed acute myocarditis during PD-1 inhibitor treatment, and the disease progressed rapidly. Case 2 was more serious than case 1. Endomyocardial biopsy showed definite cardiomyocytic injury and prominent inflammatory infiltration in both cases, which met the full Dallas criteria for myocarditis. The degenerated and necrotic cardiomyocytes accounted for about 10% of the tissues in case 1 and 30% in case 2, respectively. In case 1, the inflammatory cells counted in the densest area were about 150/HPF, comprised of CD20 + cells (about 5/HPF), CD3 + cells (about 60/HPF), CD8 + cells (about 50/HPF) and CD68 + cells (about 70/HPF). In case 2, the inflammatory cells counted in the densest area were about 350/HPF, comprised of CD20 + cells (0/HPF), CD3 + cells (about 100/HPF), CD8 + cells (about 90/HPF) and CD68 + cells (about 200/HPF). In both cases, PD-L1 + cardiomyocytes aggregated in the inflammatory lesions, and the percentage was about 8% and 30% in case 1 and case 2, respectively. Conclusions:ICI-induced myocarditis is frequently acute onset, severe symptoms, and rapid progression. The histological morphology meets the full Dallas criteria for myocarditis. Expression of PD-L1 in cardiomyocytes can be detected in the inflammatory lesions. The inflammatory cells are comprised of CD8 + T lymphocytes and macrophages and the number of macrophages significantly exceeds that of lymphocytes. Combined with the pathological characteristics and the history of ICI treatment, the diagnosis of ICI-induced myocarditis can be made.

Objective:To observe the clinical efficacy of Shenqi-Dihuang Decoction combined with conventional western medicine in the treatment of stage Ⅳ Diabetic Nephropathy (DN). Methods:Made a retrospective analysis of 80 patients of stage Ⅳ DN with the syndrome of qi and yin deficiency combined with blood stasis from June 2017 to June 2018, from Beijing Hospital of Traditional Chinese Medicine and The Third Affiliated Hospital of Beijing University of Chinese Medicine. The 80 patients were divided into treatment group and control group, 40 in each group. The control group was given the conventional western treatment. The treatment group was given Shenqi-Dihuang Decoction on the basic of the control group. Both of the treatments for the two groups lasted for 12 weeks. Before and after the treatment, the changes of TCM syndrome scores were observed in each group. The automatic biochemical analyzer was used to detect the level of fasting plasma glucose, 2-hour postprandial glucose (2 hPG), Boold Urea Nitrogen (BUN), Serum Creatinine (SCr), Plasma Albumin (Alb), Total Cholesterol (TC), Triglyceride (TG), Uric Acid (UA), Superoxide Dismutase (SOD), Homocysteine (HCY) before and after treatment. The Glycosylated Hemoglobin Alc were measured by high performance liquid chromatographic (HPLC), 24-hour urine total protein quantity (24 hUTP) were measured by immunoturbidimetric assay before and after treatment, and then Glomerular Filtration Rate (eGFR) was estimated to evaluate clinical effect. Results:After the treatment, the total effective rate in the treatment group was 87.5% (35/40), and the control group was 67.5% (27/40) and the difference was statistically significant ( χ2=4.557, P=0.033). After the treatment, the scores of urine turbid, weakness of waist and knees, fatigue, edema of both lower limbs were decreased in both groups, and the treatment group was significantly better than those of the control group ( t value were -2.178, -1.675, -3.667, -1.904, -4.835, respectively, all Ps<0.01). After the treatment, the level of FPG and HbA1c were significantly lower than those of the control group ( t value were-3.781, -8.557, respectively, all Ps<0.01); the systolic blood pressure was significantly lower than that of the control group ( t=-2.883, P=0.005); the level of TC was significantly lower than that of the control group ( t=-3.321, P=0.008); the level of Alb was significantly higher than that of the control group ( t=1.510, P=0.367); the level of 24 hUTP was significantly lower than that of the control group ( t=-2.819, P=0.008); the level of HCY were significantly lower than that of the control group ( t=-2.053, P=0.043); the level of SOD was significantly higher than that of the control group ( t=2.849, P=0.018). Conclusions:Shenqi-Dihuang Decoction combined with conventional western medicine can reduce 24 hUTP quantity of Ⅳ DN patients with the syndrome of qi and yin deficiency combined with blood stasis, reduce kidney damage, delay the development of Ⅳ DN, improve clinical effect and protect the kidney function.

OBJECTIVE： To prepare Paclitaxel（PTX）nanostructured lipid carriers （NLC） modified by small peptide alanine-glutamic acid-tyrosine-leucine-arginine （AEYLR）， and to evaluate its anti-tumor effect in vitro and in vivo. METHODS： NLC， PTX-NLC （P-NLC） and AEYLR modified P-NLC （A-P-NLC） were prepared by emulsion evaporation-low temperature solidification curing method. Its appearance， particle size， multi-dispersion index（PDI） and Zeta potential were characterized，encapsulation rate，drug loading and in vitro drug release were detected respectively. Using NCI-H1299 and S180 cells as objects， CCK-8 method was adopted to investigate inhibitory effects of free PTX， P-NLC and A-P-NLC （0.44-44.00 μg/mL， by PTX） to those cells. The half inhibition concentration （IC50） was calculated. Using S180 tumor-bearing mice as model animal， anti-tumor effects of free PTX， P-NLC and A-P-NLC （5 mg/kg， by PTX） were evaluated. RESULTS： P-NLC and A-P-NLC were round-like and dispersed evenly. The particle size， PDI and Zeta potential of A-P-NLC were （43.92±0.76） nm， 0.203±0.034 and （－19.77±1.16） mV， which were all increased to certain extent， compared with P-NLC. The encapsulation efficiency and drug loading of A-P-NLC were （95.71±0.68）％ and（1.97±0.25）％， which were both decreased to certain extent， compared with P-NLC. The cumulative release rate of A-P-NLC was（35.17±2.08）％ within 48 h， showing significant sustained-release effect compared with free PTX； the release of A-P-NLC was slower than P-NLC. Compared with free PTX and P-NLC， inhibitory rates of same concentration of A-P-NLC to NCI-H1299 cells and S180 cells were almost increased significantly， while IC50 values were all decreased significantly. There was no death in S180 tumor-bearing mice treated with A-P-NLC and the general condition was good； the volume of tumors was significantly reduced， the mass of tumors was significantly reduced， and the inhibition rate of tumors was significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS： A-P-NLC has significantly sustained-release effects； its inhibitory rate to NCI-H1299 cells and S180 cells in vitro， and its inhibitory effects on S180 solid tumor in mice are all better than free PTX and P-NLC， while the toxicity is decreased to certain extent.