Role of ASXLI loss in biological functions of HEL cells / 中国药理学通报

Aim To reveal the underlying mechanisms of the co-occurrence of ASXLI and JAK2ymr mutation by using human leukemia cell line HEL that carried homozygous /4A2V617F mutation in the elucidation of the role of ASXLI loss of function mutation in myeloproliferative neoplasms, so as to provide an important model for investigating the role of ASXLI mutation in myeloproliferative neoplasms at the cellular level. Methods HEL cell line with ASXLI knockout ( HEL-AKO) was established by using CRISPR/Cas9-mediated gene editing. And a series experiments based were preformed to verify the effect of ASXLI on HEL cell proliferation, clone formation and chemosensitivity. Results HEL-AKO cell line was successfully established, confirmed by sequencing results. We found that the loss of ASXLI could inhibit proliferation and induce cell cycle arrest at the G2/M phase. The colony-form- ing capacity of HEL-AKO cells was also markedly inhibited. Moreover, the HEL-AKO had higher cloning efficiency than HEL Control after ruxolitinib treatment. Conclusions Loss of function of ASXLI has an impact on cell biological function of HEL. Therefore, HEL- AKO cell line can be used to further explore the biological contribution of concomitant ASXLI in /4A2V617F mutated MPN.

Study on synergistic inhibitory effect of ruxolitinib and decitabine on growth of HEL cells with TET2 knockdown / 中国药理学通报

Aim To explore the cytotoxic and synergistic effects of decitabine and ruxolitinib on HEL cells with TET2 knockdown. Methods Stable TET2 knockdown by shRNA was established in HEL cell line. The change of cell proliferation was measured by CCK-8 assay. The median lethal dose (IC50) and colony formation assay were used to evaluate the cytotoxic effects of decitabine and ruxolitinib, the synergistic effects of which was further analyzed by Chou-Talalay method. Results The inhibition of TET2 increased the proliferative capacity of HEL cells. HEL cell lines became resistant to decitabine following shRNA-media- ted TET2 inactivation. Colony formation assay showed that the drug sensitivity of decitabine and ruxolitinib both decreased in TET2 knockdown HEL cells. The synergistic inhibitory effects of ruxolitinib and decitabine on TET2 knockdown HEL cells were observed. Conclusion The combination of ruxolitinib and decitabine may be an effective therapeutic strategy for accelerated or blast phase MPN patients with JAK2V6m and TET2 mutations.

Preparation of Magnetic Solid Lipid Nanoparticles Co-loaded Quercetin and Resveratrol and Evaluation of Its Antitumor Effect / 中国实验方剂学杂志

Objective:To prepare magnetic solid lipid nanoparticles co-loaded quercetin and resveratrol(QR-MSLN),develop the reasonable characterization method,and investigate its inhibitory effect on transplanted hepatocarcinoma H22 tumor in mice. Method:Magnetic Fe3O4 particles coated with oleic acid(OA-Fe3O4) were synthesized and its structure was confirmed by Fourier transform infrared spectrometer(FT-IR).QR-MSLN was prepared by emulsion ultrasonic dispersion method,its morphology was examined by transmission electron microscopy,its particle size was determined by laser particle sizer.Concentration of Fe in the preparation was determined by phenanthroline spectrophotometry.The entrapment efficiency,saturation magnetization,in vitro release behavior were investigated by ultrafiltration centrifugation,vibrating sample magnetometer(VSM) and dialysis method,respectively.Mouse tumor model transplanted with hepatoma H22 ascites tumor was established and antitumor effect of QR-MSLN on H22 bearing mice were observed in the presence of an applied magnetic field. Result:Morphology of QR-MSLN was round,and black magnetic particles could be observed inside it,its particle size was (171.9±2.2) nm,the concentration of Fe was (1.40±0.46) g·L-1.The preparation exhibited apparent superparamagnetism and the saturation magnetization was 7.75 A·m2·kg-1.The entrapment efficiencies of quercetin and resveratrol in QR-MSLN were 99.10% and 80.83%,respectively.QR-MSLN had a significantly higher effect of tumor inhibition than SLN(containing quercetin and resveratrol) and free drug(PConclusion:QR-MSLN has uniform particle size and good magnetic response,and shows remarkable antitumor effect on H22 bearing mice.

Losartan inhibits LPS-induced GFAP expression via AMPK activation in mouse hippocampus / 中国病理生理杂志

AIM:To investigate the effects of losartan on lipopolysaccharide (LPS)-induced glial fibrillary acidic protein (GFAP) expression,and to determine whether adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activation is involved in the mechanism.METHODS:Adult male KM mice were divided into control group,LPS model group,losartan treatment group,and losartan and Compound C co-treatment group.To establish a model of central nervous system inflammation,the mice received daily intracerebroventricular injection of LPS (24 μg/d) for 2 d.Daily losartan administration (0.5,1 or 5 mg · kg-1 · d-1,ip) initiated at 14 d prior to LPS injection.Compound C (10 mg/kg,ip),a selective AMPK inhibitor,started to be injected daily at 2 d prior to LPS injection.The hippocampal tissues in each group were isolated at 3 d after the last LPS injection,and then the protein levels of GFAP,AMPK,p-AMPK,mammalian target of rapamycin (mTOR) and p-mTOR were determined by Western blot.RESULTS:Twice LPS injections significantly increased the expression of GFAP in the hippocampus (P ＜ 0.01).Losartan inhibited LPS-induced GFAP expression in a concentration-dependent way,and losartan at 5 mg· kg-1 · d-1 significantly inhibited GFAP expression and AMPK activation (P ＜ 0.05),but it had no obvious effect on mTOR activation.Furthermore,Compound C significantly reversed the effect of losartan treatment on LPS-induced GFAP expression and AMPK phosphorylation (P ＜ 0.05).CONCLUSION:Losartan inhibits LPS-induced GFAP expression in the mouse hippocampus,and AMPK activation but not mTOR,is involved in the mechanism.

Losartan inhibits LPS-induced GFAP expression via AMPK activation in mouse hippocampus / 中国病理生理杂志

AIM:To investigate the effects of losartan on lipopolysaccharide (LPS)-induced glial fibrillary acidic protein (GFAP) expression,and to determine whether adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) activation is involved in the mechanism.METHODS:Adult male KM mice were divided into control group,LPS model group,losartan treatment group,and losartan and Compound C co-treatment group.To establish a model of central nervous system inflammation,the mice received daily intracerebroventricular injection of LPS (24 μg/d) for 2 d.Daily losartan administration (0.5,1 or 5 mg · kg-1 · d-1,ip) initiated at 14 d prior to LPS injection.Compound C (10 mg/kg,ip),a selective AMPK inhibitor,started to be injected daily at 2 d prior to LPS injection.The hippocampal tissues in each group were isolated at 3 d after the last LPS injection,and then the protein levels of GFAP,AMPK,p-AMPK,mammalian target of rapamycin (mTOR) and p-mTOR were determined by Western blot.RESULTS:Twice LPS injections significantly increased the expression of GFAP in the hippocampus (P ＜ 0.01).Losartan inhibited LPS-induced GFAP expression in a concentration-dependent way,and losartan at 5 mg· kg-1 · d-1 significantly inhibited GFAP expression and AMPK activation (P ＜ 0.05),but it had no obvious effect on mTOR activation.Furthermore,Compound C significantly reversed the effect of losartan treatment on LPS-induced GFAP expression and AMPK phosphorylation (P ＜ 0.05).CONCLUSION:Losartan inhibits LPS-induced GFAP expression in the mouse hippocampus,and AMPK activation but not mTOR,is involved in the mechanism.

Effects of histamine on mRNA expression of Egr-1 in astrocytes / 中国病理生理杂志

AIM:To explore whether histamine can regulate the expression of early growth response factor-1 (Egr-1) in the cerebral cortex astrocytes.METHODS:Normal wild-type (WT) mice, histidine decarboxylase knockout ( HDC-KO) mice and histamine treated HDC-KO mice were sacrificed for extracting the total RNA of the cerebral cortex. Primary cultured rat cortical astrocytes were treated with histamine at concentrations of 10 -8 , 10 -7 , 10 -6 , 10 -5 or 10 -4 mol/L for 15, 30, 60, 120 or 240 min.H1 or H2 receptor antagonists were pretreated for 15 min before histamine treat-ment.After histamine treatment, the cell total RNA or protein was extracted.The expression of Egr-1 at mRNA and protein levels was determined by real-time PCR and Western blot.RESULTS:The mRNA level of Egr-1 in cerebral cortex of HDC-KO mice was significantly lower than that in WT mice, while exogenous histamine induced the mRNA expression of Egr-1 in HDC-KO mice.In cultured astrocytes, histamine induced the mRNA expression of Egr-1.The maximum increase in the mRNA level of Egr-1 was produced by histamine at concentration of 10 -5 mol/L.In addition, histamine-induced Egr-1 mRNA accumulation peaked at 30 min after commencing stimulation, while histamine significantly increased Egr-1 protein expression at 60 min.Furthermore, histamine-induced Egr-1 expression was inhibited by H1 receptor antagonist but not H2 receptor antagonist.CONCLUSION:Histamine up-regulates the Egr-1 expression in cerebral cortex and cultured astrocytes, which may attribute to H1 receptor activation.

A wound dressing made of nanosilver and bacterial cellulose / 中国组织工程研究

BACKGROUND:In recent years, bacterial celulose modified by nano-composite technology has been endowed with new properties. OBJECTIVE:To review the combination of bacterial celulose and nanosilver to prepare wound dressing. METHODS: A computer-online search was performed in PubMed (2013-01/2015-04) and CNKI (2007-01/2015-04) databases to retrieve studies on bacterial celulose, nanosilver and their compound method and application using the key words of “bacterial celulose, nano-silver” in English and Chinese, respectively. RESULTS AND CONCLUSION:Bacterial celulose/nano-silver compound can be prepared by three methods: solution impregnation, in situ composite and biocomposite. Solution impregnation method can lower the concentration of nanosilver ions in the fiber matrix to highly control the release of silver ions, but the genetic toxicity and biocompatibility are unclear.In situcomposite method can reduce the damage to the mesh structure of celulose on which silver ions can be bonded firmly to reduce the toxic damage to cels, but the reducing agent used has a higher toxicity, which is difficult to remove. Biocomposite method cannot produce toxic substance, which is friendly to the environment, and the synthetic biomaterials have less harm to the human body and can be controled highly.

Clinical significance of early diagnosis and prognostic value of serum cell-free DNA on patients with cervical carcinoma / 中国综合临床

Objective To explore the clinical significance of the serum cell-free DNA on early diagnosis and prognostic value of patients with cervical carcinoma. Methods Forty-five patients with cervical carcinoma and 50 healthy women from May 2012 to February 2014 were selected from the Central Hospital of Ezhou. Serum cell-free DNA(cf-DNA),carbohydrate antigen125(CA125),carbohydrate antigen19-9(CA19-9), squamous cell carcinoma antigen( SCC ),β-human chorionic gonadotropin( β--HCG ) were detected by electrochemiluminescence analyzer. Results The levels of serum cf-DNA,SCC,β-HCG of patients in the cervical carcinoma group were(178. 8 ± 43. 4)μg/ L,(0. 7 ± 0. 3)μg/ L,(1. 3 ± 0. 4)kU/ L respectively, higher than those in health women((1 354. 7 ± 267. 2)μg/ L,(1. 4 ± 0. 4)μg/ L,(5. 2 ± 0. 6)kU/ L),and the differences were significant(t = 30. 690,9. 709,37. 614;P < 0. 001). Serum cf-DNA level of patients with Ⅱ-Ⅲperiod cervical carcinoma was(2 538. 8 ± 775. 3)μg/ L,higher than patients with Ⅰ period((1 244. 7 ± 253. 6)μg/ L;t = 8. 191,P < 0. 001). The area under the curve of ROC of serum cf-DNA,CA125,CA19-9, SCC,β-HCG were 0. 951 6,0. 638 2,0. 614 4,0. 805 7,0. 753 1 respectively and the AUC of serum cf-DNA was the largest. The sensibility,specificity,accuracy of cf-DNA were 75. 56% ,99. 13% ,93. 36% and 28. 32% , 93. 24% ,64. 26% in terms of CA125. Meanwhile,the sensibility,specificity,accuracy of CA19-9 were 19. 75% , 94. 47% ,67. 28% and 38. 87% ,88. 53% ,80. 16% in terms of SCC. The sensibility,specificity,accuracy of β-HCG were 43. 28% ,86. 21% ,77. 83% respectively. Conclusion Serum cf-DNA has high sensibility, specificity and accuracy in early diagnosis and prognostic value of patients with cervical carcinoma and it is worthy of clinical promotion.

Postoperative prognosis of estimated glomerular filtration rate in coronary artery bypass graft surgery / 中华检验医学杂志

Objective To evaluate the postoperative prognosis of the modification of diet in renal disease formula (MDRD) in coronary artery bypass graft surgery (CABG) in hospital or 4 years after hospitalization. Methods Two hundred and seventy-two CABG patients were divided into 3 groups according to the levels of estimated glomerular filtration rate (eGFR) including 35 cases in eGFR < 60 ml/min group, 119 cases in 60 ≤ eGFR < 90 mL/min group and 118 cases in eGFR ≥90 ml/min group. The prognostic factors of CABG patients were analyzed by COX proportional hazards models. Kaplan-Meier survival analysis was used to compare survival curves among the three groups stratified by eGFR levels. The Log-rank statistic was used for comparing between groups. Results By multivariate COX regression adjustment for body mass index, smoking, hypertension, hyperlipaemia, diabetes mellitus, previous MI, perioperative PCI and etc. , the relative risk (RR) of the increasing age for cardiac events was 1.077(95% CI 1.002-1.158,P =0.044). RR of left ventricular ejection fraction (LVEF) was 0.005(95% CI 0.000-0.456,P =0.022). RR of eGFR was 0.968(95% CI 0.948-0.988,P =0.002). The survival rate in the first, second, third and fourth year were same in every group. The survival rate of group with eGFR < 60 ml/min, 60 ≤ eGFR < 90 ml/min and eGFR≥ 90 mL/min was 76.4%, 93.1%, and 96.6%. The survival rates among three groups were statistically significant. In the survival curve of 4 year follow-up after CABG, the survival rate of group with eGFR < 60 mL/min was lower than that of 60 ≤ eGFR < 90 ml/min group and eGFR ≥ 90 ml/min group. Conclusions The preoperative eGFR is an independent risk factor in evaluating cardiac events in hospital and after hospitalization. It has a higher prognosis value in patients undergoing CABG.

Clinical characteristics of acute non-ST segment elevation myocardial infarction / 中国综合临床

Objective To study the clinical characteristics of acute non-ST segment elevation myocardial infarction(NSTEMI).Methods 211 patients of NSTEMI and STEMI underwent coronary artery angiography and echocardiogram.Patients'history and symptome were collected and the data of coronary artery angiography and echocardiogram were analyzed.Results Compared with STEMI,NSTEMI patients had more risk factors and postinfarction angina pectoris;severe coronary artery disease and three coronary vessel disease.But NSTEMI had relatively little effects on cardiac function.Conclusion NSTEMI always has more severe coronary artery disease and postinfarction ischemic effects.So more attention should be paid to its standard therapy.

Serum citrulline: a potential marker for intestinal epithelial mass and absorption capacity in short bowel syndrome patients / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the potential role of serum citrulline level in evaluating the intestinal absorptive area and capacity in patients with short bowel syndrome (SBS).</p><p><b>METHODS</b>Serum citrulline concentration was determined using high performance liquid chromatography (HPLC) in SBS patients (n=22) and healthy controls (n=33). In SBS patients, the remnant small bowel lengths and diameters were measured by radiography, and their 5- hour urine D- xylose excretion and intestinal protein absorption were also determined. The correlationship of serum citrulline level with remnant small bowel length, surface area, protein and D- xylose absorption was analyzed. The 6 patients receiving intestinal rehabilitative therapy, serum citrulline level, protein and D- xylose absorption after therapy were also measured.</p><p><b>RESULTS</b>Serum citrulline level of SBS patients was significantly lower than that of healthy controls [(5.94+/- 2.65) vs [(16.87 +/- 5.97) micromol/L, P < 0.01]. In SBS patients, serum citrulline was positively correlated with remnant small bowel length and surface area (r=0.82 and r=0.86 respectively). There was also a significant correlationship of serum citrulline level with 5- hour D- xylose excretion (r=0.56) and intestinal protein absorption (r=0.48). Serum citrulline, 5- hour D- xylose excretion and intestinal protein absorption were all significantly raised in patients after rehabilitative therapy, although no correlation of increasing percentage was found among above three parameters.</p><p><b>CONCLUSIONS</b>Serum citrulline concentration is positively correlated with intestinal absorptive area and capacity in SBS patients. It is a potential marker for evaluating the severity of intestinal failure and the efficacy of rehabilitative therapy in short bowel patients.</p>

Prophylactic treatment with growth hormone improves intestinal barrier function and alleviates bacterial translocation in stressed rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Damage to the gut barrier often occurs during critical illnesses. In such cases, it is very important to alleviate impairment of the intestinal barrier and protect intestinal barrier function. This study investigated the protective effect of growth hormone on intestinal barrier function in rats under stress.</p><p><b>METHODS</b>This study consisted of prospective, randomized, and controlled animal experiments. Twenty-five Sprague-Dawley rats served as total parenteral nutrition (TPN) models and were divided into three groups: TPN group, sepsis (Sep) group, and growth hormone (GH) group. Another 8 rats served as normal controls. Each group received different stress stimuli. Rats were fed for 7 days, and samples were taken for examination 24 hours after gavaging with dual saccharides.</p><p><b>RESULTS</b>The architecture of the small intestinal mucosa in the Sep group showed the most severe damage among all groups. Nitric oxide levels in blood plasma and immunoglobulin A levels in the intestinal mucosa of the GH group were significantly lower than in the Sep group (P < 0.02). There were no significant changes in CD3 counts and in the CD4/CD8 ratio between the four groups. Dual sugar tests and bacteriological examinations revealed that intestinal permeability and rate of bacterial translocation in the GH group were lower than in the Sep group (P < 0.01, respectively).</p><p><b>CONCLUSION</b>Prophylactic treatment with growth hormone can alleviate damage to intestinal barrier function caused by trauma and endotoxemia in rats under stress.</p>

Combination chemotherapy with etoposide and cisplatin for high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia / 中华妇产科杂志

Objective To evaluate the feasibility and effectiveness of combination chemotherapy with etoposide and cisplatin(EP)regimen on the patients with high-risk,chemorefractory and recurrent gestational trophoblastic neoplasia(GTN).Methods Thirty-nine patients with gestational trophoblastic tumors were analyzed retrospectively,25 of 39 patients were of high-risk,9 patients were chemorefractory and 5 patients were recurrent.All 39 patients were administrated with EP regimen,and 10 patients were assisted with surgery.All the patients were followed up.Clinical response,toxicity,the occurrence of secondary tumors of all patients,and the fertility of 30 patients whose fertility function was preserved were investigated. Results Thirty-nine GTN patients underwent a total of 221 cycles of the EP regimen.The average number of courses for each patient was 5.7.The total complete remission rate of the regimen was 74%(29/39). Twenty-five patients with high-risk GTN received a total of 139 cycles and the average number of courses was 5.6.Nineteen patients achieved complete remission and 6 patients showed drug-resistant.The complete remission rate of the high-risk group was 76%(19/25).Nine patients with chemorefractory GTN obtained a total of 55 cycles and the average number of courses was 6.1.Six patients achieved complete remission and 3 patients showed drug-resistant again.The complete remission rate of the chemorefractory group was 6/9. Five patients with recurrent GTN received 27 cycles and the average number of courses was 5.4.Four patients achieved complete remission,1 patient showed drug-resistance and died.Bone marrow toxicity, gastrointestinal reaction and alopecia were the main side effects of the EP regimen,but the bone marrow toxicity was slight and no grade Ⅳ side effect occurred.No fatal effect was found.Eight of 30 patients whose fertility faction was preserved had become pregnant after recovery,with a total of 8 pregnancies.Among them,2 were terminated by induced abortion,and 6 underwent normal term delivery and gained 6 infants who had no congenital malformation.All the 6 children had normal growth and development after childbirth. None of the women developed secondary tumors.Conclusion The EP regimen is effective and safe for the treatment of high-risk,chemorefractory and recurrent GTN.

CLINICAL OBSERVATION AND ANALYSIS OF BLOOD COAGULATION IN PATIENTS WITH CYANOTIC CONGENITAL HEART DISEASE / 解放军医学杂志

Platelet count (PC) and plasma prothrombin time (PPT) of sixty- six patients with cyanotic congenital heart disease (cyanotic CHD) were determined, and linear relationship analysis was made between PC, PPT and hemoglobin (HB), Hemotocrit saturation of blood (SaO2). Blood viscosity (BV) and hemotocrit (HT) were also examined. It was discovered that there was a significant positive linear relationship between SaO2 and PC, HB and PPT, and a significant negative linear relationship between PC and HB, BV, HT, respectively. There was a significant difference of PC in cyanotic-CHD (n = 66) as compared with noncyanotic-CHD (t=13.9508, P

Changes of QT dispersion in electrocardiogram after coronary artery bypass graft / 第二军医大学学报

Objective:To study the significance of QT dispersion(QTd) changes in electrocardiogram(ECG) after coronary artery bypass graft. Methods:Forty eight patients scheduled for elective coronary artery bypass graft were studied. The changes of QTd were monitored by 12 lead electrocardiogram system before surgery (as baseline values), at the end of surgery, and 1 4 d and 7 d after surgery.Results:The baseline values for QTd was (56.32?2.25) ms.QTd were significantly longer as compared to the baseline values ( P