Site-Specific Anchoring a Luminescent Tag in a Protein with Non-Emissive Iridium(III) Complex.

Site-specific modification of proteins with synthetic fluorescent tag effectively improves the resolution of imaging, and such a labeling method with negligible three-dimensional structural perturbations and minimal impact on the biological functions of proteins is of high interest to dissect the high-resolution activities of biomolecules in complex systems. To this end, several non-emissive iridium(III) complexes [Ir(C-N)2 (H2 O)2 ]+ OTF- (C-N denotes various cyclometalated ligands) were designed and synthesized. These complexes were tested for attaching a protein by coordinating to H/X (HisMet, HisHis, and HisCys) that are separated by i and i+4 in α-helix. Replacement of the two labile water ligands in the iridium(III) complex by a protein HisHis pair increases the luminescent intensity up to over 100 folds. This labeling approach has been demonstrated in a highly specific and efficient manner in a number of proteins, and it is also feasible for labeling target proteins in cell lysates.

ALDH2, ADCY3 and BCMO1 polymorphisms and lifestyle-induced traits are jointly associated with CAD risk in Chinese Han people.

BACKGROUNDS: To investigate associations of genetic and environmental factors with coronary artery disease (CAD), we collected medical reports, lifestyle details, and blood samples of 2113 individuals, and then used the polymerase chain reaction (PCR)-ligase detection reaction (LDR) to genotype the targeted 102 SNPs. METHODS: We adopted elastic net algorithm to build an association model that considered simultaneously genetic and lifestyle/clinical factors associated with CAD in Chinese Han population. RESULTS: In this study, we developed an all covariates-based model to explain the risk of CAD, which incorporated 8 lifestyle/clinical factors and a gene-score variable calculated from 3 significant SNPs (rs671, rs6751537 and rs11641677), attaining an area under the curve (AUC) value of 0.71. It was found that, in terms of genetic variants, the AA genotype of rs671 in the additive (adjusted odds ratio (OR) = 2.51, p = 0.008) and recessive (adjusted OR = 2.12, p = 0.021) models, the GG genotype of rs6751537 in the additive (adjusted OR = 3.36, p = 0.001) and recessive (adjusted OR = 3.47, p = 0.001) models were associated with increased risk of CAD, while GG genotype of rs11641677 in additive model (adjusted OR = 0.39, p = 0.044) was associated with decreased risk of CAD. In terms of lifestyle/clinical factors, the history of hypertension (unadjusted OR = 2.37, p < 0.001) and dyslipidemia (unadjusted OR = 1.82, p = 0.007), age (unadjusted OR = 1.07, p < 0.001) and waist circumference (unadjusted OR = 1.02, p = 0.05) would significantly increase the risk of CAD, while height (unadjusted OR = 0.97, p = 0.006) and regular intake of chicken (unadjusted OR = 0.78, p = 0.008) reduced the risk of CAD. A significantinteraction was foundbetween rs671 and dyslipidemia (the relative excess risk due to interaction (RERI) = 3.36, p = 0.05). CONCLUSION: In this study, we constructed an association model and identified a set of SNPs and lifestyle/clinical risk factors of CAD in Chinese Han population. By considering both genetic and non-genetic risk factors, the built model may provide implications for CAD pathogenesis and clues for screening tool development in Chinese Han population.

[CLAG Regimen Composed of Continuous Intravenous Infusion of Cladribine in the Treatment of Refractory/Relapsed Acute Myeloid Leukemia].

OBJECTIVE: To study the efficacy and safety of continuous intravenous infusion of 2-Chlorodeoxyadenosine (2-CdA) combined with high-dose cytarabine (Ara-C) and granulocyte colony-stimulating factor (G-CSF) (CLAG regiem) in the treatment of relapsed/refractory acute myeloid leukemia (AML). METHODS: Fifteen patients with refractory/relapsed AML hospitalized in 5 medical units such as Department of Hematology, the Affiliated Tumor Hospital of Zhengzhou University and received one course of CLAG regimen from June 2014 to August 2019 were analyzed retrospectively (specifically: cladribine 5 mg/M2, day 1 to day 5, continuous 24-hour intravenous infusion; Ara-C 2 g/M2, 1 time/day, day 1 to day 5, intravenous infusion; G-CSF 300 mg, 1 time/day, day 0 to day 5, subcutaneous injection). RESULTS: Among the 15 patients with refractory/relapsed AML, 9 males and 6 females, the median age was 35 (13-63) years old. FAB classification: 1 case of M1, 3 cases of M2a, 4 cases of M2b (including 1 case with extramedullary invasion), 1 case of M4 with extramedullary invasion, 5 cases of M5, 1 case of HAL; NCCN classification: 6 cases in intermediate risk group, 9 cases in high risk group; 8 cases refractory, 7 cases relapsed. The median time of pre-chemotherapy was 4 (2-8) (of which NO.15 had received 8 cycles of chemotherapy and received CLL1-CAR-T), and the median white blood cell count before chemotherapy was 12.27 (from 0.78 to 5.29)×109/L. After 1 course of treatment with CLAG regimen, 12 patients achieved complete remission (12/15, 80%), and the median duration of CR was 65 days (0-528) days. IV grade leukopenia and thrombocytopenia was found in all the patients after chemotherapy. The median duration of granulocytosis was 20 (14 to 33) days, and 1 patient died. Seven patients received allogeneic hematopoietic stem cell transplantation. The median EFS and OS time of 15 patients was 85 (19-558) days and 117 (19-558) days, respectively. CONCLUSION: The CLAG regimen consisting of continuous intravenous infusion of cladribine shows high CR in the treatment of AML patients, but the duration of CR is short, myelosuppression is sever, so that infection control is the key. Allogeneic hematopoietic stem cells transplantation should be performed as soon as possible after CR.

[Synergistic Mechanism of Interferon alpha-1b, Interleukin-2 and Thalidomide for Immune Regulation in Patients with Acute Myeloid Leukemia].

OBJECTIVE: To explore the synergistic immunomodulatory mechanism of interferon alpha-1b, interleukin-2 and thalidomide (ITI) regimen on patients with acute myeloid leukemia (AML). METHODS: Sixty eight untreated de novo or relapsed or refractory or maintenance therapy patients with AML admitted in the Affiliated Cancer Hospital of Zhengzhou University and the other 11 medical units from March 2016 to May 2019 were treated with ITI regimen. Peripheral blood specimen per patient was collected into EDTA-K3 anticoagulation vacuum tube before the administration of ITI and 3 months after the treatment; peripheral blood lymphocyte subsets and perforin and Granzyme B expression were analyzed by using flow cytometry; the levels of VEGF, IFN-γ, TNF-α and IL-6 in the plasma were detected by using a cytometric bead array. Thirty-five healthy subjects from the hospital physical examination centre were selected as normal controls. RESULTS: The ratio of CD4+/CD8+ T cells, the percentage of NK cells, the expression of perforin and Granzyme B of NK cells in the peripheral blood of patients with hematological malignancies were lower than those of healthy controls. The level of VEGF, IL-6 and TNF-α in the peripheral plasma were higher than those of the healthy control group, and the difference was statistically significant. The level of IFN-γ was lower, and the difference was not statistically significant. The ratio of CD4+/CD8+ T cells, the percentage of NK cells, the expression of Granzyme B and Perforin of NK cells in peripheral blood were higher after the therapy of thalidomide combined with rhIFNα-1b for 3 months as compared with those before treatment of ITI, the level of the IFN-γ in peripheral plasma was higher while that of VEGF was lower, the difference was statistically significant; after treatment, the ratio of CD3+ CD4+ and CD3+ CD8+ lymphocytes and the level of TNF-α in peripheral blood were higher those that before treatment, IL-6 was lower, while the difference was not statistically significant. CONCLUSION: The ITI regimen can raise the ratio of CD4+/CD8+ T cells and the percentage of natural killer cells, also, can enhance the generation of perforin and granzyme B and the concentration of IFN-γ as well as inhibit the generation of VEGF, suggesting that these activities may enhance the antitumour capacity of patients with AML.

Effects of bronchial thermoplasty and cryoablation on airway smooth muscle / 中华医学杂志(英文版)

BACKGROUND@#The effectiveness of bronchial thermoplasty (BT) has been reported in patients with severe asthma. This study compared the effects of BT and cryoballoon ablation (CBA) therapy on the airway smooth muscle (ASM).@*METHODS@#Eight healthy male beagle dogs were included in this experiment. In the preliminary experiment, one dog received BT treatment for both lower lobe bronchus, another dog received CBA treatment for 7 s on the upper and lower lobe of right bronchus, and 30 s on the left upper and lower lobe. The treatments were performed twice at an interval of 1 month. In subsequent experiments, the right lower lobe bronchus was treated with BT, and the left lower lobe bronchus was treated with CBA. The effects of treatment were observed after 1 (n = 3) month and 6 months (n = 3). Hematoxylin-eosin staining, Masson trichrome staining, and immunohistochemical staining were used to compare the effects of BT and CBA therapy on the ASM thickness, collagen fibers synthesis, and M3 receptor expression after treatment. One-way analysis of variance with Dunnett post hoc test was used to analyze the differences among groups.@*RESULTS@#In the preliminary experiment, the ASM ablation effect of 30-s CBA was equivalent to that of 7-s CBA (ASM thickness: 30.52 ± 7.75 μm vs. 17.57 ± 15.20 μm, P = 0.128), but the bronchial mucociliary epithelium did not recover, and large numbers of inflammatory cells had infiltrated the mucosal epithelium at 1-month post-CBA with 30-s freezing. Therefore, we chose 7 s as the CBA treatment time in our follow-up experiments. Compared with the control group (35.81 ± 11.02 μm), BT group and CBA group (13.41 ± 4.40 μm and 4.81 ± 4.44 μm, respectively) had significantly decreased ASM thickness after 1 month (P < 0.001). Furthermore, the ASM thickness was significantly lower in the 1-month post-CBA group than in the 1-month post-BT group (P = 0.015). There was no significant difference in ASM thickness between the BT and CBA groups after six months (9.92 ± 4.42 μm vs. 7.41 ± 7.20 μm, P = 0.540). Compared with the control group (0.161 ± 0.013), the average optical density of the ASM M3 receptor was significantly decreased in 6-month post-BT, 1-month post-CBA, and 6-month post-CBA groups (0.070 ± 0.022, 0.072 ± 0.012, 0.074 ± 0.008, respectively; all P < 0.001). There was no significant difference in the average optical density of ASM M3 receptor between the BT and CBA therapy groups after six months (P = 0.613).@*CONCLUSIONS@#CBA therapy effectively ablates the ASM, and its ablation effect is equivalent to that of BT with a shorter onset time. A neural mechanism is involved in both BT and CBA therapy.

[Analysis of the Irregular Antibodies of the Patients and Its Clinical Significance].

OBJECTIVE: To retrospectively analyze the identification results of irregular antibodies, to clarify the distribution features and to explore the relation of alloantibodies and autoantibodies with the immunized history of patients and disease kinds. METHODS: 49 820 patients who applied for red blood transfusion during Sep 1st 2017 to Sep 1st 2018 were selected. All the specimens were screened for the antibody by microcolumn gel antiglobulin technique, which then were identified for irregular antibody. RESULTS: Antibodies were found in 861 (1.73%) of all 49 820 transfused samples. The alloimmunization history of the patients with antibodies was significantly different between male and female (χ2=18.54，P＜0.01). The alloantibody was the most common, accounting for 59.50% in all of the antibodies. Warm autoantibody, anti-E, anti-M, anti-cE and anti-Ce accounted for 68.5% of the antibodies. The blood group of Rh, MNS and Lewis were responsible for 92.40% of alloantibody, especially anti-E accounted for the largest percentage(38.60%) of alloantibody. Patients with alloantiboies experienced much more the alloimmunization and transfusion history (χ2=20.13，P＜0.01；χ2=5.40，P＜0.05) . The distribution of auto and alloantibody was very significantly different among the ddifferent isease (χ2=51.8，P＜0.01), Hematopathy, solid tumor and osteoarthropathy were often associated with alloantibody, otherwise, autoantibodies often occurred in hematopathy and autoimmune disease. CONCLUSION: The most important factor that results in antibody-screening positive is alloantibody, in which anti-E antibody from Rh blood group system in most common.

Efficacy of ixazomib for the treatment of relapsed/refractory multiple myeloma: A protocol of systematic review and meta-analysis.

BACKGROUND: Over the past years, ixazomib has been increasingly explored for the treatment of relapsed/refractory multiple myeloma (RRMM). However, its results are still contradictory. This study will explore the efficacy and safety of ixazomib for patients with RRMM. METHODS: A systematic records search of Cochrane Library, PUBMED, EMBASE, CINAHL, ACMD, PsycINFO, WANGFANG, and China National Knowledge Infrastructure will be carried out from their origin to March 31, 2020 with no limitations of language and publication status. Trials will be selected by titles/abstracts, and full manuscripts by 2 independent authors. Data collection will be carried out from eligible trials based on the previous designed criteria. Study quality will be checked using Cochrane risk of bias, and statistical analysis will be administered by RevMan 5.3 software. RESULTS: This study will summarize the current high-quality trials investigating the efficacy and safety of ixazomib for the treatment of patients with RRMM. CONCLUSION: The results of this study may provide convinced evidence on the evidence-based medicine level, and guidance for clinical practice and future studies. INPLASY REGISTRATION NUMBER: INPLASY202040027.

[Relationship between Depression and Quality of Life in Patients with Severe Obesity].

Objective To investigate the depression status and quality of life (QoL) of patients with severe obesity who have undergone bariatric surgery and to evaluate their potential correlation.Methods A total of 50 patients with severe obesity who had undergone elective bariatric surgery in the general surgical ward of Peking Union Medical College from January 2013 to December 2017 were enrolled in this study. The general data were collected. The patient's depression status and QoL were assessed by the Zung's Depression Self-Assessment Scale and the SF-36 Quality of Life Scale,respectively.Results The depression score was (44.85±9.07) scores in these 50 patients,including 38 patients (76%) without depression and 12 (24%) with mild depression. The scores of the mental-emotional,physical,psychomotor,and psychological barrier dimensions of patients with severe obesity were (3.70±1.25),(18.65±4.10),(4.25±1.35),and (18.75±5.46)scores,respectively. The physiological function score of patients with mild depression was (95.83±10.21)scores,which was significantly higher than those without depression (68.42±38.95) scores(F=6.209,P=0.011). The vitality and mental health scores were (46.67±14.72) and (57.33±14.68) scores in patients with mild depression,which were significantly lower than those [(65.79±21.94) (F=4.867,P=0.030) and (75.16±17.21) scores (F=0.212,P=0.032)] in patients without depression. Correlation analysis showed that the mental-emotional score was positively correlated with physical score in patients with severe obesity (r=0.503,P=0.015),whereas the depression score was negatively correlated with the general health status (r=-0.430,P=0.032),vitality (r=-0.594,P=0.002),social functioning (r=-0.455,P=0.022),mental health (r=-0.562,P=0.003),and QoL score (r=-0.439,P=0.028).Conclusions The QoL is remarkably affected by depression in patients with severe obesity before surgery. Assessment of the mental status of patients with severe obesity may help to identify this problem and thus improve the QoL.

[Current status of blood pressure control in patients with coronary heart disease].

OBJECTIVE: To evaluate the level of blood pressure control in patients with coronary heart disease (CHD) of China in order to provide guidance for the prevention and treatment of CHD. METHODS: The patients with CHD were retrospectively collected from 2011~2014 in PLA General Hospital and Hainan Branch Hospital. Then analyzed the difference of blood pressure compliance rate between different surgical methods percutaneous coronary intervention (PCI), coronary artery bypass grafting(CABG), secondary preventive drugs(aspirin, clopidogrel, nitrates, trimetazidine, nicorandil, hypotensor, hypoglycemic, lipid-lowering drugs) and lifestyle(smoking, drinking, exercise). RESULTS: ①Effects of surgical methods on blood pressure:Male's systolic blood pressure (SBP) and diastolic blood pressure(DBP) in the CABG group were lower in the PCI group and control group, and female's DBP in the CABG group were lower in the PCI group. ②Usage rate of secondary prevention drugs:usage rate of trimetazidine, calcium antagonist, ß-blockers, angiotensin-converting enzyme inhibitor (ACEI) and angiotensin receptor antagonist (ARB) in hypertension group were higher than in normal blood pressure group. ③ Lifestyle condition:compliance rate of blood pressure in the smoking group was lower than that in the non smoking group. And there was no significant difference in blood pressure compliance rate among whether drinking and doing exercise or not. CONCLUSIONS: Blood pressure control in patients with CHD was still not satisfied. Compared with PCI, CABG may be more beneficial in the control of blood pressure in patients with CHD. Smoking cessation and improving the usage rate of secondary preventive drugs are still the main means of blood pressure control.

Different combination strategies for prophylaxis of venous thromboembolism in patients: A prospective multicenter randomized controlled study.

The aim was to evaluate the efficacy and safety of different combination strategies for prophylaxis of venous thromboembolism (VTE) after gynecologic surgery in patients at different levels of risk. This was a prospective multicenter randomized controlled study, in which 625 women who would undergo pelvic surgery for gynecologic diseases were stratified into three risk groups and then randomized into four groups to receive graduated compression stockings (GCS) alone (group A), GCS + low molecular weight heparin (LMWH) (group B), GCS + intermittent pneumatic compression (IPC) (group C), and GCS + IPC + LMWH (group C), respectively. The overall incidence of DVT was 5.1%. Group A had the highest incidence of DVT (8.8%), followed by group C (5.2%), group B (3.8%), and group D (2.6%). There was a significant difference in the incidence of DVT between groups A and D. The incidence of DVT was significantly lower in LMWH-treated patients (group B + group D) than in non-LMWH-treated patients (group A + group C). In conclusion, combination prophylaxis, especially LMWH-containing strategies, is better than monoprophylaxis in reducing VTE after gynecologic surgery. Risk-stratified prophylactic strategies should be implemented in patients undergoing gynecologic surgery, with LMWH-containing strategies being recommended for high-risk and very-high-risk patients.

Successful treatment of accessory breast cancer with endocrine therapy.

Accessory breast cancers in males are extremely rare, and only a few cases have been reported in the literature. In this paper, an 87-year-old male patient was diagnosed with an accessory breast cancer by means of computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography-computed tomography (PET-CT), and immunohistochemistry based on needle biopsy, and has undergone successful resection and postoperative adjuvant endocrine therapy. He was the oldest male patient with an accessory breast cancer reported in the Chinese Hospital Knowledge Database and PubMed literature from 1975 to 2015.

The future of remote ECG monitoring systems.

Remote ECG monitoring systems are becoming commonplace medical devices for remote heart monitoring. In recent years, remote ECG monitoring systems have been applied in the monitoring of various kinds of heart diseases, and the quality of the transmission and reception of the ECG signals during remote process kept advancing. However, there remains accompanying challenges. This report focuses on the three components of the remote ECG monitoring system: patient (the end user), the doctor workstation, and the remote server, reviewing and evaluating the imminent challenges on the wearable systems, packet loss in remote transmission, portable ECG monitoring system, patient ECG data collection system, and ECG signals transmission including real-time processing ST segment, R wave, RR interval and QRS wave, etc. This paper tries to clarify the future developmental strategies of the ECG remote monitoring, which can be helpful in guiding the research and development of remote ECG monitoring.

Application and Exploration of Big Data Mining in Clinical Medicine.

OBJECTIVE: To review theories and technologies of big data mining and their application in clinical medicine. DATA SOURCES: Literatures published in English or Chinese regarding theories and technologies of big data mining and the concrete applications of data mining technology in clinical medicine were obtained from PubMed and Chinese Hospital Knowledge Database from 1975 to 2015. STUDY SELECTION: Original articles regarding big data mining theory/technology and big data mining's application in the medical field were selected. RESULTS: This review characterized the basic theories and technologies of big data mining including fuzzy theory, rough set theory, cloud theory, Dempster-Shafer theory, artificial neural network, genetic algorithm, inductive learning theory, Bayesian network, decision tree, pattern recognition, high-performance computing, and statistical analysis. The application of big data mining in clinical medicine was analyzed in the fields of disease risk assessment, clinical decision support, prediction of disease development, guidance of rational use of drugs, medical management, and evidence-based medicine. CONCLUSION: Big data mining has the potential to play an important role in clinical medicine.

Mitochondrial tRNA mutations in patients with myelodysplastic syndromes.

Increasing evidence showed that mitochondria play an important role in the development of myelodysplastic syndromes (MDS). Mitochondrial dysfunctions caused by mitochondrial DNA mutations, especially mitochondrial tRNA mutations, were found to be associated with MDS in many studies. However, the link between a candidate mitochondrial tRNA mutation and MDS was not clear. In this study, we investigated the role of some mitochondrial tRNA mutations, and their deleterious roles were further discussed.

Effect of immune modulation therapy on cardiac function and T-bet/GATA-3 gene expression in aging male patients with chronic cardiac insufficiency.

AIM: The aim of this study was to explore the role of immune modulation therapy in regulating the imbalance of Th1/Th2, serum IFN-γ, IL-4 and the T-cell-specific transcription factors T-bet/GATA-3 in peripheral blood in aging male patients with chronic cardiac insufficiency (CCI). PATIENTS & METHODS: In total, 156 participants were divided into three groups: the CCI intervention group, which received regular therapy and thymopetidum (20 mg intramuscular injection, once every other day for 3 months; n = 70), the CCI control group, which received regular therapy (n = 56) and 50 healthy individuals older than 57 years of age, who served as normal controls. RESULTS: Before therapy, in comparison with the control group, levels of left ventricular end diastolic diameter, NT-proBNP, C-reactive protein (CRP), Th1, Th1/Th2, IFN-γ, and T-bet mRNA and T-bet/GATA-3 mRNA all increased, and the level of left ventricular ejection fraction (LVEF), 6MWT, Th2, IL-4, and GATA-3 mRNA also decreased in both the CCI intervention and control groups. Linear correlation analysis indicated that LVEF was inversely correlated with serum NT-proBNP, CRP, Th1/Th2, IFN-γ and T-bet mRNA/GATA-3 mRNA, and was positively correlated with plasma IL-4. After 3 months of therapy, levels of left ventricular end diastolic diameter, NT-proBNP, CRP, Th1, Th1/Th2, IFN-γ, T-bet mRNA and T-bet/GATA-3 mRNA decreased in the two CCI subgroups, but levels in the CCI intervention group were lower in comparison to the control group. Levels of LVEF, 6MWT, Th2 and GATA-3 mRNA increased in the two CCI subgroups, while levels in the CCI intervention group were higher in comparison with the control group. Plasma levels of IL-4 showed no change after treatment. CONCLUSION: Immune modulation improved cardiac function of CCI patients and was associated with amelioration of T-helper superficial transcription factor polarization and its related cytokine imbalance. Immune modulation might be a new treatment strategy for aging CCI patients.

Two mechanical methods for thromboembolism prophylaxis after gynaecological pelvic surgery: a prospective, randomised study.

BACKGROUND: Venous thromboembolism is known to be an important social and health care problem because of its high incidence among patients who undergo surgery. Studies on the mechanical prophylaxis of thromboembolism after gynaecological pelvic surgery are few. The aim of our study was to evaluate the effect of mechanical thromboembolism prophylaxis after gynaecological pelvic surgery using a combination of graduated compression stockings (GCS) and intermittent pneumatic compression (IPC) or GCS alone. METHODS: The study was performed on 108 patients who were randomly assigned to two groups. The first group received GCS before the operation and IPC during the operation (IPC + GCS group). The second group received GCS before the operation (GCS group). To analyze the effect of the preventive measures and the laboratory examination on the incidence of thrombosis and to compare the safety of these measures, the incidence of adverse reactions was assessed. RESULTS: The morbidity associated with DVT was 4.8% (5/104) in the IPC + GCS group and 12.5% (14/112) in the GCS group. There were significant statistical differences between the two groups. There were no adverse effects in either group. CONCLUSIONS: The therapeutic combination of GCS and IPC was more effective than GCS alone for thrombosis prevention in high-risk patients undergoing gynaecological pelvic surgery, and there were no adverse effects in either group.

[Effects of matrix metalloproteinase-9 inhibitor in Lewis rats with experimental autoimmune myocarditis].

OBJECTIVE: To investigate the effects of matrix metalloproteinase-9 (MMP-9) inhibitor minocyclin hydrochloride in Lewis rats with experimental autoimmune myocarditis (EAM). METHODS: EAM was induced by injection of cardiac C protein emulsified in completed Freund adjuvant in double footpad and intraperitoneal injection of pertussis toxin on 6- to 8-week old Lewis rats. Sixty EAM Lewis rats were divided into 3 groups (early, middle and late intervention groups, n = 20 each: 10 minocyclin treated and 10 control rats). In early intervention group, rats in treatment group received intraperitoneal injection of minocyclin hydrochloride from 1(st) to 21(st) day after immunization; in middle intervention group, rats were treated from 8(th) to 28(th) day after immunization and in late intervention group, rats were treated from 15(th) to 35(th) day after immunization (50 mg/kg body weight, once daily). Control rats received intraperitoneal injection of same volumetric physiological saline at corresponding time periods. At the end of intervention, rats were euthanatized and hearts were harvested. Paraffin sections were used for hematoxylin and eosin stain to determine the inflammatory score, for picrosirius stain to determine fibrosis score and collagen content, and for immunohistological stain to determine macrophages and T lymphocytes. Real time PCR was used to detect mRNA expression of myocardial MMP-2 and MMP-9. Cryostat sections were used for in situ zymography to detect protein activity of gelatinase. RESULTS: Inflammatory score in cardiac paraffin slides, number of cardiac macrophages and T lymphocytes, cardiac interstitial fibrosis score and content, expression of MMP-2, 9 mRNA and activity of gelatinase in treatment group were all significantly lower than in control group for early and middle intervention groups (inflammatory score: early control group vs. treatment group: 3.03 ± 1.35 vs.1.51 ± 0.36, P < 0.05, middle control group vs. treatment group: 3.75 ± 0.29 vs. 2.11 ± 0.82, P < 0.01; cardiac interstitial fibrosis score, early control group vs. treatment group: 2.75 ± 0.29 vs.1.51 ± 0.35, P < 0.01, middle control group vs. treatment group: 2.50 ± 0.41 vs. 1.61 ± 0.42, P < 0.05; gelatinase, early control group vs. treatment group: 162 367 ± 5095 vs. 62 366 ± 2131, P < 0.01, middle control group vs. treatment group: 184 256 ± 5427 vs. 113 197 ± 4809, P < 0.01) while these parameters were similar between minocyclin-treated and control rats in late intervention group (all P > 0.05). CONCLUSIONS: MMP-9 plays an important role in the pathogenesis of autoimmune myocarditis. Inhibition of MMP-9 in early and middle stage could significantly attenuate inflammatory responses and myocardial fibrosis in this experimental EAM model.

[Immune state of Th1, Th2 and Th17 subpopulation in experimental autoimmune myocarditis].

OBJECTIVE: To shed light on changes in the gene expression of T helper lymphocyte (Th) subpopulation, Th1, Th2 and Th17 in autoimmune myocarditis and to gain insight into the immunological mechanisms underlying the essence of myocarditis. METHODS: An experimental Lewis rat autoimmune myocarditis model was induced by immunization with cardiac C protein and completed Freund adjuvant in double foot pads and subsequent intraperitoneal injection of pertussis toxin. Two groups of normal rats without immunological injection acted as control group. Transthoracic echocardiography was performed and subsequently hearts and spleens were obtained from EAM rats at 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks after immunization. The pathological sections of heart samples were prepared, the inflammatory score was determined by hematoxylin and eosin stain, the fibrosis score was determined by picrosirius red stain. The ratio of Th1, Th2 and Th17 subpopulation in spleen cells were measured by flow cytometry, and enzyme linked immunoabsorption assay (ELISA) was used to determine the serum level of Th1 related cytokine interferon-gamma (IFN-gamma), Th2 related cytokine interleukin (IL)-4 and Th17 related cytokine IL-17. RESULTS: In EAM rats, cardiac ejection fraction remained normal until 4th week, and left ventricular end systolic diameter and left ventricular end diastolic diameter decreased. However, cardiac ejection fraction decreased obviously and left ventricular end systolic diameter and left ventricular end diastolic diameter rose until 8th week. Inflammatory score increased rapidly at 2nd week after immunization and remained peek level until 4th week and then gradually decreased at 6th and 8th week. Fibrosis score and fibrosis content increased from 4th week and maintained the peek level until 8th week. Ratio of Th1 in spleen cells of EAM rats and its related cytokine in serum, IFN-gamma, increased at 1st week, arrived at the peek level until 4th week and gradually decreased at 6th week. Ratio of Th17 and IL-17 rose from 2nd-4th week and remained until 6th to 8th week. Ratio of Th2 showed no change in the previous four weeks, ratio of IL-4 increased from 4th week, and both rose at 6th week rapidly and remained until 8th week. CONCLUSIONS: In EAM Lewis rats, The time duration from 2nd to 4th week was inflammatory stage of myocarditis while during the period of 4th week to 8th week myocarditis develops into fibrotic stage. Imbalance of Th1/Th2 takes part in the occurrence of ventricular remodeling, cellular immunity mediated by Th1 and Th17 being preponderant at inflammatory myocarditis stage while humoral immunity mediated by Th2 and Th17 being preponderant at fibrotic carditis stage.

[Clinical significance of preventive treatment of thrombosis for patients undergoing gynecological surgery with high risk factors].

OBJECTIVE: To explore the clinical significance of preventive treatment of thrombosis for patients undergoing gynecological surgeries with high risk factors. METHODS: Prospectively, randomized and cases controlled study was performed in 143 patients underwent gynecological surgery with high risk factors to explore the prevention effectiveness and safety in morbidity of lower extremity deep venous thrombosis (LDVT) and relative effected factors, who were enrolled to three groups according to different preventive treatment: group using intermittent pneumatic calf compression (IPC) in 47 cases, group using low-molecular-weight hepairin (LMWH) in 48 cases and control group in 48 cases with no prevention strategy. RESULTS: The morbidity of LDVT on lower extremity was 6% (6/94) in group IPC and 1% (1/96) in group LMWH, 18% (17/96) in control group, which was obviously reduced in group IPC and group LMWH than that in control group (P < 0.05), while there was no significantly difference between group IPC and group LMWH (P > 0.05). After surgery in 3 or 4 days, the morbidity of LDVT in group IPC was 1/6, zero in group LMWH and 71% (12/17) in control group, there was significantly reduced in group IPC and group LMWH than that in control group (P < 0.05). There was no side-effects in group IPC, only 1 case in group LMWH presented small quantity bleed in vagina remnant, while no incision bleeding and bleed tendency. Single-variate analysis indicated that elder age, abdominal surgeries and malignant tumor were as the independent factors to effect preventive treatment of thrombosis (P < 0.05). CONCLUSION: LDVT in patients underwent gynecological surgeries with high risk factors can be obviously reduced and delayed by preventive treatment, which is no side-effects, while be effected by the factors of elder age, abdominal surgeries and malignant tumor.