Objective To induce and identify the differentiation of rat bone marrow mesenchymal stem cells(MSCs)into cardiomyocytes in vitro,and observe the expression of Nesprin protein during the differentiation. Methods Rat MSCs were isolated and purified by Ficoll density gradient centrifugation,and adhered for serial subcultivation.Surface-associated antigens of MSCs of the second passage were dedected by flow cytometry.MSCs of the second passage were induced by 10μmol/L 5-azacytidine(5-Aza)to differentiate into cardiomyocytes,and the morphological changes were observed.The expression of Desmin,α-sarcomeric actin and cardiac Troponin I(cTnI) mRNA and protein was detected by RT-PCR,immunocytochemistIv and immunofluorescence staining, and the expression of Nesprin protein was detected by Western blotting. Results The morphology of MSCs induced by 5-Aza was bigger and longer,and the nuclei became bigger,exhibiting more consistent patterns.The expression of Desmin,α-sarcomeric actin and cTnI mRNA and protein of MSCs was positive.Immunofluorescence staining revealed that Nesprin protein positioned in the nuclear membrane,and Western blotting detection demonstrated that the expression of Nesprin protein significantly increased after differentiation(P<0.05).Conclusion MSCs may be successfully induced to differentiate into cardiomyocytes.The expression of Nesprin protein in the differentiated MSCs may significantly increase,indicating Nesprin may play a role in the differentiation from MSCs to cardiomyocytes.

Objective: To observe the impact of cardiac contractility modulation (CCM) on myocardial remodeling in rabbit model of chronic heart failure (CHF) with its possible mechanism. Methods: Rabbit HF model was established by ascending aortic root ligation; the animals were divided into 3 groups: Sham group, the animals received thoracotomy without aortic ligation, HF group and HF+CCM group, the HF animals received CCM treatment for 4 weeks. n=10 in each group. Cardiac function was measured by echocardiography at 12 and 16 weeks in each group respectively; myocardial tissue fibrosis and pathological changes were examined by Masson staining; plasma BNP level was assessed by ELISA; protein expressions of collagen I, collagen II, MMP2,MMP9, TIMP1 and galectin-3 in myocardial tissue were determined by Western blot analysis. Results: ① By echocardiography: with 12 weeks treatment, compared with Sham group, HF group and HF+CCM group had increased LVESD, LVEDD and decreased LVFS, LVEF, all P<0.05; with 16 weeks treatment, compared with HF group, HF+CCM group had improved LVESD, LVEDD, LVEF and LVFS, all P<0.05. ② Pathological changes:compared with Sham group, HF group showed increased collagen content in myocardial tissue, P<0.05; CCM treatment could partially decrease collagen accumulation, P<0.05. ③ After 12 weeks treatment, compared with Sham group, HF group and HF+CCM group presented elevated plasma BNP level, P<0.05; after 16 weeks treatment, compared with HF group, HF+CCM group presented reduced plasma BNP, while it was still higher than that in Sham group, P<0.05. ④ By Western blot analysis: compared with Sham group, HF group demonstrated increased protein expressions of collagen I, collagen II, MMP2, MMP9, TIMP1 and galectin-3 in myocardial tissue; the above indexes were much lower in HF+CCM group while still higher than those in Sham group, all P<0.05. Conclusion: CCM could improve myocardial remodeling in rabbit model of CHF which might be related to down-regulated protein expressions of collagen I, collagen III, MMP2, MMP9, TIMP1 and galectin3 in myocardial tissue.

OBJECTIVETo investigate the characteristics and clinical significance of changes in gamma delta T cells in patients with chronic hepatitis B (CHB), including during the immune tolerant (IT) phase and the immune activated (IA) phase.

METHODSFlow cytometry was used to analyze the frequencies and absolute numbers of γδT cells and their subsets in peripheral blood and in liver from 80 CHB cases, including 20 IT carriers and 60 IA patients. Blood samples were obtained from all CHB cases and 5 healthy controls (HCs). Liver biopsies were collected from 22 IA patients and 5 IT carriers undergoing diagnosis, and from the 5 HCs.

RESULTSCompared to HCs and IT carriers, the IA patients displayed significantly lower levels of peripheral and intrahepatic γδT cells as well as the Vδ2 subsets. The levels of peripheral and intrahepatic VγδT cells were closely associated with the liver histological activity index and serum alanine aminotransferase levels.

CONCLUSIONγδT cells, especially the Vδ2 subsets, may play a protective role in decreasing liver damage in CHB patients.

Alanine Transaminase ; blood ; Case-Control Studies ; Flow Cytometry ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Liver Cirrhosis ; immunology ; T-Lymphocyte Subsets ; cytology

Objective: To detect the expression of CRLF2 in adult Ph negative acute B lymphocytic leukemia (B-ALL) in newly diagnosed cases, and to investigate the relationship between CRLF2 and the general clinical characteristics, efficacy and prognosis. Methods: 103 cases of newly diagnosed adult B-ALL patients were investigated from Apr 2016 to Dec 2017 in the Department of Hematology, Henan Cancer Hospital. Bone marrow samples was used to detect the expression of CRLF2 in leukemic cells. The expression of CRLF2 ≥20% was defined as CRLF2-high group and <20% was defined as CRLF2-low group. The clinical characteristics and prognosis of the two groups were compared. Results: The Median overall survival (OS) and disease free survial (DFS) in CRLF2-high group were 9.0 months and 4.25 months, respectively. CRLF2-low group were 15.5 months and 10.25 months, respectively. There was a statistically significant difference in median OS and DFS between the two groups (P=0.007, P=0.000) . The 18-month OS and DFS in CRLF2-high group were 38.6% and 25.1%, respectively. CRLF2-low group were 57.8% and 42.3%, respectively. Multivariate analysis showed high expression of CRLF2 was an independent risk factor for OS (HR=2.991, 95% CI 1.429-6.261, P=0.004) and DFS (HR=2.374, 95%CI 1.146-4.960, P=0.041) in patients. Conclusion Patients with high expression of CRLF2 had poor prognosis.

Objective:To explore the value of susceptibility weighted imaging (SWI) in evaluating renal excess iron deposition.Methods:Thirty New Zealand white rabbits were randomly divided into iron group (injected iron dextran) with 15 rabbits and control group with 15 rabbits. All rabbits underwent SWI examination at 0th week before and 8th week after the injection of iron. MRI images of the control and iron group at the 0th and 8th week were analyzed. The phase value of renal cortex was measured on the phase images and the angle radian value was calculated. The distribution of renal iron deposition was observed by Prussian blue stain. The renal iron content was measured by atomic absorption spectrophotometer. The Mann-Whitney U or Wilcoxon test was used to compare the difference in angular radian value between the two groups. The independent sample t test was used to compare the difference in renal iron content between the two groups. Results:Only in iron group at 8th week, the SWI signal intensity of renal cortex was significantly lower than that of medulla. The renal cortical SWI signal intensity of iron group at 8th week was significantly lower than that of iron group at 0th week and control group at 8th week, respectively. The angle radian value of iron group at 8th week [0.202 3(0.161 8, 0.272 5)] was significantly higher than that of iron group at 0th week [-0.045 4 (-0.013 9, 0.008 0)] and control group at 8th week [-0.011 2 (-0.052 9, -0.001 4)], respectively ( Z=-3.408, -4.666, all P<0.05). Only in the iron group at 8th week, the obvious blue iron deposition were observed in renal cortex and few were observed in medulla. The renal iron content [(135.3±14.1) mg/kg] of iron group at 8th week was significantly higher than that [(75.5±9.8) mg/kg] of control group at 8th week ( t=13.938, P<0.001). Conclusions:SWI can evaluate the excess iron deposition in kidney. Excess iron is mainly deposited in the renal cortex and reduces its SWI signal intensity. The angle radian value calculated from the phase value can quantify the iron deposition.

Objective:To explore the value of susceptibility-weighted imaging (SWI) in quantitative evaluation of iron load in diabetic kidneys.Methods:Thirty two healthy New Zealand white rabbits were randomly divided into diabetic group (DM, n = 20) and control group (NC, n = 12). DM model was established by injecting 5% alloxan solution (100 ml/kg) through ear vein. 12 rats were finally enrolled into the group. NC group was injected with the same dose of normal saline. DM group and NC group were intramuscularly injected with 60 mg/kg iron dextran. The left kidney was scanned by MRI immediately after iron injection (0 weeks) and 12 weeks after feeding. The left kidney was killed after 12 weeks of scanning. The left kidney was examined by Prussian blue staining and atomic absorption spectrophotometer. The value of SWI in quantitative evaluation of renal iron content was evaluated by using the iron content measured by atomic absorption spectrophotometer as the gold standard. On SWI phase diagram, the region of interest (ROI) was manually drawn along the renal cortical vagal area, and the measured phase values were converted into angular radians. Mann Whitney U test was used to compare the blood glucose value and the angle radian value at 0 week and 12 week between the two groups; independent sample t test was used to compare the difference of iron content between the two groups; nonparametric Wilcoxon signed rank test was used to compare the difference of angle radian between DM group and NC group at 0 and 12 weeks; Spearman correlation analysis was used to study the correlation between angle radian value and atomic absorption spectrophotometer results. Results:The blood glucose level in DM group [28.0 (10.6) mmol/L] was significantly higher than that in NC Group [6.5 (1.9) mmol/L], and the difference was statistically significant (U = 0, P<0.001). At week 0, there was no significant difference between DM group [-0.04 (-0.02)] and NC Group [-0.02 (0.06)] in angle radian value (U=105.50, P>0.05); at 12 weeks, the angle radian value of DM group [0.22 (0.17)] was higher than that of NC Group [0.17 (0.05)], the difference was statistically significant (U=35.50, P<0.05). The angle radian of DM group and NC group at 12 weeks were higher than that of 0 weeks, and the differences were statistically significant ( P<0.05). Prussian blue staining showed that iron was mainly deposited in renal cortex, and the blue staining in DM group was more obvious than that in NC group. The signal intensity of renal cortex on SWI images in DM group was significantly lower than that in 0 week group at 12 weeks, and slightly decreased in NC group. The iron content of DM group and NC group were (171.39±20.13) mg/kg and (116.21±28.90) mg/kg, respectively, and the difference was statistically significant ( t=5.428, P<0.001). Spearman correlation analysis showed that the angle radian was positively correlated with iron content ( r=0.67, P<0.001). Conclusions:Diabetic kidneys have more iron deposits than normal kidneys. As a non-invasive, simple and convenient examination technique, SWI has the potential to quantitatively evaluate the iron load of diabetic kidneys.

With the course of Clinical Skill Training as an example, this article integrates hierarchical teaching and peer teaching, adopts the problem-based learning (PBL) teaching model and famous traditional Chinese medicine (TCM) teaching clinics, connects the assessment criteria for standardized residency training, and pays attention to cultivating a reasonable teacher team, so as to improve the overall teaching quality of the course and the enthusiasm for the course among students. The application of the teacher inheritance model combined with case-based learning (CBL) teaching can help medical students to learn and inherit the clinical experience and skills, communication methods, and academic ideas of famous old TCM doctors and inherit and carry forward the essence of TCM culture. At the same time, CBL teaching is adopted to promote the integrated and people-oriented teaching concept of TCM, with medical students as the main body and teachers as the supporting role. The training with this model can effectively improve the clinical skills and development potential of TCM students.

Objective:To explore the expression of programmed death ligand-1 (PD-L1) protein in grade Ⅲ non-special type invasive breast cancer and its related factors, so as to provide a basis for immunotherapy.Methods:A total of 63 surgically resected specimens of grade Ⅲ non-special type invasive breast cancer diagnosed by pathology in Cancer Hospital & Shenzhen Hospital of Chinese Academy of Medical Sciences from February 2017 to February 2021 were collected. The HE-stained sections were reviewed, and the proportion of immune cells (IC) in all invasive active tumor cells in the tumor sections was calculated. The expression of PD-L1 (SP142) protein in all specimens was detected by immunohistochemistry. The relationship between the positive expression of PD-L1 protein and the clinicopathological parameters was analyzed, and the Pearson correlation test was used to analyze the relationship between the expression of PD-L1 (SP142) and the degree of IC infiltration.Results:Among 63 patients, 19 patients (30.2%) were triple-negative type, 34 patients (53.9%) were luminal type, and 10 patients (15.9%) were human epidermal growth factor receptor 2 (HER2) overexpression type. The positive rate of PD-L1 (SP142) in grade Ⅲ non-special type invasive breast cancer was 77.8% (49/63). The positive rate of PD-L1 (SP142) in triple-negative type was 94.7% (18/19), the positive rate in non-triple-negative type was 70.5% (31/44), and the difference between the two groups was statistically significant ( P = 0.047). The positive rate of PD-L1 (SP142) in estrogen receptor (ER)-negative patients or progesterone receptor (PR)-negative patients was both 90.3% (28/31), which was higher than that of ER-positive patients or PR-positive patients (65.6%, 21/32), and the difference was statistically significant ( P = 0.018); the positive rate of PD-L1 (SP142) was not related to the patient's age, tumor site, tumor maximum diameter and number, vascular tumor thrombus, nerve invasion, lymph node metastasis, HER2 status and Ki-67 positive index (all P > 0.05). The expression of PD-L1 was positively correlated with the degree of IC infiltration ( r = 0.716, P<0.001). Conclusion:In grade Ⅲ non-special type invasive breast cancer, the patients with triple-negative type has a high positive rate of PD-L1, and the patient with negative ER or PR has a high positive rate of PD-L1; the tumor IC infiltration is positively correlated with the expression of PD-L1.

Objective:To investigate outcomes and safety of doxycycline-moxifloxacin sequential regimen in the treatment of Mycoplasma genitalium urethritis/cervicitis. Methods:From June 2019 to December 2020, patients with Mycoplasma genitalium urethritis/cervicitis confirmed by nucleic acid amplification testing were successively recruited at Department of Sexually Transmitted Diseases, Hospital of Dermatology, Chinese Academy of Medical Sciences, and received sequential therapy with oral doxycycline for 7 days followed by oral moxifloxacin for 7 days. Clinical and/or etiological assessment was conducted 2 to 3 weeks after the end of treatment. Fisher′s exact test was used to analyze factors influencing the treatment outcome. Results:Totally, 36 eligible subjects were enrolled, including 30 males and 6 females. Among them, 18 (50%) patients completed post-treatment etiological assessment, which showed that 12 achieved microbiological cure, and treatment failures occurred in 6; another 18 patients achieved clinical cure. The overall response rate to doxycycline-moxifloacin sequential therapy was 83.3% (30/36, 95% confidence interval[ CI]: 70.5%, 96.1%) . The treatment outcome showed no significant association with the patients′ age, gender, marital status, number of sexual partners in the past 1 month, history of sexually transmitted diseases, history of antibiotic use in the past 1 month, or co-infections (all P > 0.05) . Conclusion:The efficacy of doxycycline-moxifloacin sequential regimen is limited in the treatment of Mycoplasma genitalium infections in Nanjing area, and clinicians should be alerted to the possibility of treatment failure in clinical practice.