Objective To explore the mechanism of azithromycin (AZM) for inhibiting the proliferation of rat airway smooth muscle cells (ASMCs).Methods Thirty Sprague-Dawley (SD) rats were divided into the control group,asthma model group and AZM group.The rat model of asthma was established by ovalbumin (OVA) sensitization and stimulation in vitro.The airway related parameters of rat lung tissue were determined by using the medical image analysis system.Primary passage ASMCs were isolated and cultured using the tissue-sticking method,and the vascular endothelial growth factor (VEGF) overexpression vector or tumor necrosis factor receptor-associated factor 6 (TRAF6) overexpression vector was transfected into ASMCs in the AZM group.The protein levels of VEGF,NF-κB p65 and TRAF6 were detected by Western blotting,and the proliferation of ASMCs was evaluated by CCK-8 kit.Results AZM significantly inhibited the increase of thickness of total airway wall,thickness of inner airway wall and thickness of airway smooth muscle layer in asthma rats (P<0.05),also significantly inhibited the proliferation of ASMCs in the asthma model group (P<0.05).AZM significantly inhibited the protein expression of VEGF and NF-κB p65 induced by asthma (P<0.05),and the overexpression of VEGF significantly reduced the inhibiting effects of AZM on proliferation of ASMCs (P<0.05).AZM significantly inhibited the high expression of TRAF6 induced by asthma (P<0.05),and the overexpression of TRAF6 significantly reduced the inhibiting effects of AZM on expression of VEGF and NF-κB p65 as well as proliferation of ASMCs (P<0.05).Conclusion AZM can suppress the proliferation of ASMCs,its partial mechanism may be realized through inhibiting TRAF6/NF-κB/VEGF signaling pathway.

AIMTo explore the changes of mRNA and protein expressions of heart-type fatty acid binding protein (H-FABP) in rat ischemic myocardium at different intervals ischemia.

METHODS60 SD male rats weighing 250-350 g, were randomly divided into one sham-operated group and five study groups (group A1, A2, A3, A4, A5, the left coronary artery of rats has been ligated for 1 h, 2 h, 4 h, 6 h, 12 h respectively). Myocardil samples from infarct zone, ischemic and non-ischemic zone, were obtained for histology examination, and the mRNA for H-FABP in ischemic myocardial tissue were determined by RT-PCR. Serum free fatty acid(FFA) was determined by colorimetric method.

RESULTSCompared to sham hearts, H-FABP mRNA expression were significantly decreased in ischemia zone of AMI rat hearts (P < 0.05), especially in rats underwent 4 h ischemia and 6 h ischemia (P < 0.01). Serum FFA were significantly increased in AMI rats relative to sham rats (P < 0.05).

CONCLUSIONSignificant down-regulated heart-type fatty acid binding protein after myocardial ischemia might play an important role in myocardial injury and energy metabolism disorder.

Animals ; Down-Regulation ; Fatty Acid-Binding Proteins ; genetics ; metabolism ; Male ; Myocardial Infarction ; metabolism ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Most main oilfields in China have already entered a "double high" development stage (high water cut, high recovery degree). To further enhance oil recovery in reservoirs after polymer flooding (RAPFs), an efficient activator formulation for promoting metabolism of endogenous microorganism was studied by aerogenic experiments, physical simulation experiments, electron microscopy scanning and pyrophosphate sequencing. Results show that the activator could activate the endogenous microorganisms in the injected water and make the pressurized gas reach 2 MPa after 60 d static culture of the activator in a high pressure vessel. The oil recovery efficiency of natural core physical simulation flooding can be improved by more than 3.0% (OOIP) in RAPFs when injected 0.35 PV activator with 1.8% mass concentration, and a lot of growth and reproduction of activated endogenous microorganism in the core was observed by electron microscopy scanning. Field trial with 1 injector and 4 producers was carried out in the east of south II block of Sa Nan in December 2011. By monitoring four effective production wells, changes of carbon isotope δ13C (PDB) content of methane and carbon dioxide were -45 per thousand to -54 per thousand and 7 per thousand to 12 per thousand. Compared with east II of Sa Nan block, the oil amount increased by 35.9%, water cut stabled at 94%. The incremental oil was 5 957 t during the three and a half years, which provides an alternative approach for further improving oil recovery in similar reservoirs.
Carbon Dioxide ; chemistry ; Carbon Isotopes ; analysis ; China ; Diphosphates ; chemistry ; Methane ; chemistry ; Oil and Gas Fields ; microbiology ; Polymers ; Water ; Water Microbiology

Objective To investigate the change of adiponectin (AD),tumor necrosis factor-alpha (TNF-α) and S100 levels in serum elderly patients with rheumatoid arthritis and cerebral infarct in order to evaluate the therapeutic effect of dioscornin.Methods One hundred patients with rheumatoid arthritis and cerebral infarct were selected as our subjects,who were hospitalized in the Department of Neurology of Recovery Changzhi Municipal People's Hospital and the Department of Internal Medicine of the Affiliated Hospital of Shaoxing Medical College,between 2006 September and 2010 September.All subjects (male 55,female 45,average age of 57 years old) were randomly divided into regular and dioscomin groups,50 for per group.Patients in regular group were treated with routine therapy and patients in dioscornin groups were treated with dioscornin 80mg,three daily plus regular treatment drug.Meanwhile 40 middle patients with single rheumatoid arthritis subjects were severed as controls.The changes of BMI,fasting plasma glucose,lipid factors,insulin sensitivity index (ISI),serum adiponectin,TNF-α and serum S100B were determined at treatment before and 6 months after treatments.Results The level of TNF-α,serum S100 in ERA patients were significantly higher andAdiponectin significantly lower than that of the control group,(TNF-α:[(89.0 ± 25.3) ng/L,(88.0 ± 24.2)ng/L vs(74.0 ±21.0) ng/L,F =3.292,P ＜0.05],[S100B:(0.102 ±0.051) μg/L,(0.101 ±0.045) μg/L vs(0.092 ± 0.031) μg/L,F =2.792,P ＜ 0.05],and AD and BMI were lower [AD:(7.2 ± 1.4) μg/L,(7.3 ±1.4) μg/L vs (18.1 ± 3.5) μg/L,F =17.057,P ＜ 0.01],[BMI:(18.9 ± 2.4) kg/m2,(19.0 ± 1.9) kg/m2 vs (21.8 ± 1.8) kg/m2,F =6.147,P ＜ 0.01].There was a negative correlation between adiponect and TNF-α,S100B (r =-0.46,-0.52,P ＜ 0.01) and positive correlation between adiponect and BMI (r =0.44,P ＜0.01).The adiponectin level was significantly increased in patients for six months after dioscornin treatment than that of control group.[AD:(12.2±2.9) μg/L,(7.8 ±1.8) μg/L vs (18.0 ±4.3) μg/L,F=6.480,P＜0.01].The level of TNF-α and S100B significantly decreased than that of the control group,TNF-α:[(72.0 ±21.0) ng/L,(82.0±23.0)ng/L vs (68.0 ±20.0) ng/L,F =3.065,P ＜0.05],[S100B:(0.092 ±0.021)μg/L,(0.099 ±0.031) μg/L vs (0.091 ±0.029) μg/L,F=3.030,P＜0.05].Conclusion Dioscornin could ameliorate the prognosis through decreasing the levels of TNF-α and S100B,and increasing adiponectin level in patients with rheumatoid arthritis and cerebral infarct.

Objective: To investigate the effect of natural active compounds apigenin (API) on the proliferation of rat aortic vascular smooth muscle cells (VSMCs) induced by lipopolysaccharide (LPS) and related mechanisms. Methods: VSMCs of primary cultured SD rats were obtained and the cytotoxic effects of API (0, 10, 20, 40 and 80 μmol/L) was explored by CCK-8 method. Impact of LPS (0, 0.1, 1, 10 and 100 μg/ml) on VSMCs proliferation and the impact of API (0, 10, 20, 40 μmol/L) on LPS (10 μmol)-induced VSMCs proliferation by CCK-8 methods. Using EdU and FCM method, we observed the effect of API on proliferation of VSMCs induced by LPS. VSMCs proliferation and cell cycle were also assessed by EdU method and FACS in 10 μg/ml LPS, 10 μg/ml LPS+ 40 μmol/L API and equal volume DMSO treated VSMCs. Results: (1) CCK-8 cell vitality test showed that cell vitality was not affected by 0-40 μmol/L API, while cell vitality was significantly reduced by 80 μmol/L API (57%), which was significantly lower than in blank group (P<0.05). (2) VSMCs proliferation was significantly promoted by 0.1, 1 and 10 μg/ml LPS and peaked in 10 μg/ml LPS stimulated VSMCs group, while VSMCs proliferation was significantly reduced in 100 μg/ml LPS stimulated group (P<0.05 vs. blank group). (3) LPS (10 μm/ml) induced VSMCs proliferation was not affected by 10 μmol/L API, which was significantly inhibited by 20 and 40 μmol/L API (both P<0.05 vs. LPS). (4) VSMCs proliferation assessed by EdU was significantly higher in LPS group than in blank group (P<0.01), which could be significantly reduced by cotreatment with API (P<0.01). (5) FACS results showed that percent of VSMCs in G0/G1 stage was significantly lower in LPS group compared to blank group (P<0.05), which could be significantly increased post API treatment (P<0.05 vs. LPS), while percent of VSMCs in S stage was significantly lower post API treatment in comparison with LPS group. Conclusion API can significantly inhibit LPS-induced proliferation of VSMCs, partly through inhibiting mitosis and inducing G0/G1 cell cycle arrest.

Objective:To summarize the case data of endometrial cancer (EC), analyze the related factors of lymph node metastasis, and establish the prediction model, so as to provide reference for clinical practice.Methods:191 patients with endometrial cancer who were diagnosed and treated in department of gynecology of Baoding Maternal and Child Health Hospital from January 2010 to December 2019 were selected as the research objects. The demographic and surgical pathological information of the patients were analyzed retrospectively. The risk factors of lymph node metastasis were analyzed by univariate and logistic regression analysis, and the predictive model was established.Results:A total of 191 patients with EC, aged 26-76(53.1±9.5)years old, body mass index (BMI)18.70-40.20(25.84±3.94)kg/m 2, 13 cases (6. 81%) had lymph node metastasis. Univariate analysis showed that lymph node metastasis was associated with obesity (BMI≥28 kg/m 2), pathological type (non endometrioid adenocarcinoma), degree of differentiation, depth of myometrial invasion (>1/2) and vascular invasion ( P<0.05). Logistic multivariate analysis showed that low differentiation ( OR=9.475, 95% CI: 1.840-48.799), vascular invasion ( OR=6.614, 95% CI: 1.457-30.024) and deep muscle invasion ( OR=4.997, 95% CI: 1.342-18.600) were independent risk factors ( P<0.05). The regression equation: Logit P=-4.488+ 1.609× myometrial infiltration depth+ 1.889×vascular infiltration+ 2.249×degree of tissue differentiation. The area under the receiver operating characteristic (ROC) curve (AUC) of EC lymph node metastasis probability P was 0.813 (95% CI: 0.688-0.938). The cut off value of 0.56 was ideal. At this time, the prediction sensitivity was 76.9% and the specificity was 79.2%. Conclusions:In clinical practice, gynecologists should consider the condition of EC patients and make operation plan to avoid over treatment or under treatment.

Objective To explore the damage mechanism of dopamine cells induced by amphetamine (AMPH). Methods The damage model of dopaminergic cells in mice was established by intraperitoneal injection of AMPH. The mice were randomly grouped into control, saline, amphetamine treatment for 1 day, 7 days, 14 days and 28 days. Each group contained 10 mice. The model of cell injury was established by use of AMPH in PC12 cells. The dopaminergic fibers of corpus striatum and PC12 cells were observed by the immunohistochemistry and immunofluorescence method, and changes of proteins in the protein kinase B (Akt) / glycogen synthase kinase 3β(GSK-3β) / collapsin response mediator protein 2 (CRMP-2) signal pathway were detected by Western blotting. Results AMPH caused the damage of dopaminergic fibers in the mouse corpus striatum and PC12 cells. Meanwhile, AMPH inhibited Akt and GSK-3β phosphorylation levels, and increased phosphorylated CRMP-2 level. Nerve growth factor(NGF), an agonist of Akt, or SB216763, an inhibitor of GSK-3β protected PC12 cells against AMPH-induced toxicity through upregulation of Aat and GSK-3β phosphorylation and downregulated of phosphorylation CRMP-2. Conclusion AMPH causes damage of dopamine cells via inhibition of Akt/ GSK-3β/ CRMP-2 signal pathway.

Objective To establish the cardiac arrest (CA) model in rats by modified transcutaneous electrical stimulation on epicardium. Methods This study was performed in the Emergency Medicine laboratory in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. After 10 Sprague-Dawley male rats weighing 330-380 g were anesthetized, two acupuncture needles connected to the anode and cathode of a stimulator were transcutaneously inserted into the epicardium as electrodes. The puncture points were located quantitatively according to the anatomical structure of the rat chest. The electrical stimulation was maintained for 3 minutes to induce ventricular fibrillation(VF). Cardiopulmonary resuscitation (CPR) included chest compressions, intravenous adrenaline and defi brillation operated at 6 min after a period of nonintervention. Results CA was induced after the implement of the effective electrical stimulation in all ten rats in this experiment. The average current intensity to induce VF was (1.80 ± 0.59) mA, the average time to induce CA was (5.07 ± 2.37)s,the average time of the total electrical stimulation was(187.50 ± 12.75)s and the total time of CA was 6 min. At the end of the electrical stimulation, 9 rats presented VF and 1 rat showed pulseless electrical activity. The restoration of spontaneous circulation was achieved in all 10 rats. The average time of CPR was(190.90±68.60) s, the mean numbers of defi brillation were(1.20 ± 0.63) , and he average number of adrenaline application were (1.20 ± 0.42) times. Neither visible hemorrhage on epicardium nor gross pulmonary congestion was observed. Conclusions The modified transcutaneous electrical stimulation on epicardium to produce CA model in rats is an easily applicable and effective technique. This model may provide an alternative for experimental research of CPR.

BACKGROUND: Breast cancer patients with ipsilateral supraclavicular lymph node metastasis (ISLNM) but without distant metastasis are considered to have a poor prognosis. This study aimed to develop a nomogram to predict the overall survival (OS) of breast cancer patients with ISLNM but without distant metastasis. METHODS: Medical records of breast cancer patients who received surgical treatment at the Affiliated Cancer Hospital of Zhengzhou University, Jiyuan People's Hospital and Huaxian People's Hospital between December 21, 2012 and June 30, 2020 were reviewed retrospectively. Overall, 345 patients with pathologically confirmed ISLNM and without evidence of distant metastasis were identified. They were further randomized 2:1 and divided into training (n = 231) and validation (n = 114) cohorts. A nomogram to predict the probability of OS was constructed based on clinicopathologic variables identified by the univariable and multivariable analyses. The predictive accuracy and discriminative ability were measured by calibration plots, concordance index (C-index), and risk group stratification. RESULTS: Univariable analysis showed that estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), human epidermal growth factor receptor 2-positive (HER2+) with Herceptin treatment, and a low axillary lymph node ratio (ALNR) were prognostic factors for better OS. PR+, HER2+ with Herceptin treatment, and a low ALNR remained independent prognostic factors for better OS on multivariable analysis. These variables were incorporated into a nomogram to predict the 1-, 3-, and 5-year OS of breast cancer patients with ISLNM. The C-indexes of the nomogram were 0.737 (95% confidence interval [CI]: 0.660-0.813) and 0.759 (95% CI: 0.636-0.881) for the training and the validation cohorts, respectively. The calibration plots presented excellent agreement between the nomogram prediction and actual observation for 3 and 5 years, but not 1 year, OS in both the cohorts. The nomogram was also able to stratify patients into different risk groups. CONCLUSIONS In this study, we established and validated a novel nomogram for predicting survival of patients with ISLNM. This nomogram may, to some extent, allow clinicians to more accurately estimate prognosis and to make personalized therapeutic decisions for individual patients with ISLNM.
Breast Neoplasms ; Female ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Nomograms ; Retrospective Studies

ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang （BWT） on non-alcoholic fatty liver disease （NAFLD） and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups： normal control， model， positive drug （pioglitazone hydrochloride 1.95×10^-3 g·kg^-1）， and low-， medium-， and high-dose BWT （1.3，2.5 and 5.1 g·kg^-1）. Following a 12-week high-fat diet （HFD） inducement， the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week， glucose tolerance （GTT） and insulin tolerance （ITT） tests were conducted. Subsequently， the mice were euthanized for the collection of liver tissue and serum， and the subcutaneous adipose tissue （iWAT） and epididymal adipose tissue （eWAT） were weighed. Serum levels of total triglycerides （TG） and liver function indicators，such as alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， were determined. Histological examinations， including oil red O staining， hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy， were performed to evaluate hepatic lipid deposition， pathological morphology， and ultrastructural changes， respectively. Meanwhile， Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were employed to analyze alterations， at both gene and protein levels， the insulin signaling pathway molecules， including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 （IRS1/2/Akt/FoxO1）， glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase （Pepck） and glucose-6-phosphatase （G6Pase）， lipid metabolism-related genes stearoyl-coA desaturase-1 （SCD-1） and carnitine palmitoyltransferase-1 （CPT-1）， fibrosis-associated molecules α-smooth muscle actin （α-SMA）， type Ⅰ collagen （CollagenⅠ）， and the fibrosis canonical signaling pathway transforming growth factor-β₁/drosophila mothers against decapentaplegic protein2/3（TGF-β₁/p-Smad/Smad2/3）， inflammatory factors such as interleukin（IL）-6， IL-8， IL-11， and IL-1β， autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1， and the expression of mammalian target of rapamycin （mTOR）. ResultCompared with the model group， BWT reduced the body weight and liver weight of NAFLD mice（P<0.05， P<0.01）， inhibited liver lipid accumulation， and reduced the weight of white fat： it reduced the weight of eWAT and iWAT（P<0.05， P<0.01） as well as the serum TG content（P<0.05， P<0.01）. BWT improved the liver function as reflected by the reduced ALT and AST content（P<0.05， P<0.01）. It improved liver insulin resistance by upregulating IRS2， p-Akt/Akt， p-FoxO1/FoxO1 expressions（P<0.05）. Besides， it improved glucose and lipid metabolism disorders： it reduced fasting blood glucose and postprandial blood glucose（P<0.05， P<0.01）， improved GTT and ITT（P<0.05， P<0.01）， reduced the expression of Pepck， G6Pase， and SCD-1（P<0.01）， and increased the expression of CPT-1（P<0.01）. The expressions of α-SMA， Collagen1， and TGF-β₁ proteins were down-regulated（P<0.05， P<0.01）， while the expression of p-Smad/Smad2/3 was downregulated（P<0.05）， suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6， IL-8， IL-11 and IL-1β（P<0.01） and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression（P<0.05）while downregulating the expression of p62/SQSTM1 and mTOR（P<0.05）. ConclusionBWT ameliorates NAFLD by multifaceted improvements， including improving IR and glucose and lipid metabolism， anti-inflammation， anti-fibrosis， and enhancing autophagy. In particular， BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.