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 investigate the predictive value of lactate concentration within 1 h after admission combined with lactate clearance rate (LC) at 6 h after fluid resuscitation in prognosis of neonatal septic shock.Methods:In this retrospective study, 58 newborns with septic shock admitted to the Neonatal Intensive Care Unit of Xi′an Children′s Hospital,Xi′an Jiao Tong University from June 2016 to March 2020 were enrolled. According to the mortality within 60 days after admission,which was also set as the end point, the patients were divided into death group and survival group. The general demographic data and clinical variables including blood cell counts, procalcitonin, C-reactive protein, D-dimer, serum creatinine, and lactate concentration within 1 h after admission (Lac1) and at 6 h after fluid resuscitation (Lac2) were collected. The differences in the clinical variables between the survival and death group were compared by independent sample t test or Rank-Sum test, and the risk factors of poor prognosis were analyzed by binary Logistic regression. The predictive values of these risk factors were tested by receiver operating characteristic (ROC) curve. Furthermore, the cut-off of the risk factors were used to analyze the accumulative survival rate by Kaplan-Meier curve. Results:A total of 58 neonates were enrolled, among whom 24 survived and 34 died within 60 days after admission. The rate of premature rupture of membranes in the death group was higher than that in the survival group (41% (14/34) vs.13%(3/24), P=0.021). There were also significant differences in infection site, pathogenic characteristics, total fluid volume of resuscitation, vasoactive drug index, rate and complications of mechanical ventilation between the two groups (all P<0.05). The levels of Lac1, Lac2, procalcitonin, D-dimer and serum creatinine in the death group were higher than those in the survival group ((12±6) vs. (7±4) mmol/L, (14±6) vs. (4±2) mmol/L, (59±23) vs.(24±14) ng/L, (24±11) vs.(11±6) mg/L, (167±31) vs.(92±23) μmol/L, t=3.549, 3.112, 3.859, 4.499, 3.288, all P<0.05). While the blood pressure and LC at 6 h after fluid resuscitation were lower than those in the survival group ((41±12) vs. (52±5) mmHg (1 mmHg=0.133 kPa), t =4.230;-16 (-40, 20) % vs. 40 (18, 70) %, Z= 3.558, all P<0.05). Binary Logistic regression analysis showed that LC was negatively associated with the risk of death in neonates with septic shock (odds ratio ( OR) and 95% confidence interval ( CI): 0.679 (0.662-0.999), P<0.05), while Lac1 was the risk factor and positively associated with the risk of death ( OR and 95% CI: 1.203 (0.965-1.500), P<0.05). Furthermore, the predictive values of LC, Lac1 and the combination of these two variables in the prognosis of neonatal septic shock were analyzed by ROC curve analysis, and the area under the curve (AUC) were 0.699, 0.875, 0.965, respectively, with the sensitivity of 83.32%, 89.65% and 94.31%, and the specificity of 72.52%, 77.18% and 88.76%, respectively. According to the cut-off value of Lac1, the newborns with Lac1>4 mmol/L had significantly lower accumulative survival rate than those with Lac1≤4 mmol/L by Kaplan-Meier analysis (21% (8/38) vs. 80% (16/20), χ 2=54.520, P<0.05). According to the cut-off value of LC, the newborns with LC ≤ 10% had significantly lower accumulative survival rate than those with LC>10% by Kaplan-Meier analysis (19% (6/32) vs. 69% (18/26), χ 2=14.140, P<0.05). Conclusion:The combination of lactate concentration and lactate clearance rate have an optimal predictive value in the prognosis of neonatal septic shock.

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.

ObjectiveTo investigate the intervention effects of Suanzaoren Tang on depression model rats induced by isolation combined with chronic unpredictable mild stress （CUMS）， and to examine its influence on the c-Jun N-terminal kinase （JNK）/proto-oncogene protein （c-Myc）/tumor suppressor protein 53 （p53） signaling pathway， thereby revealing its potential functional mechanism. MethodsA total of 72 male SD rats were randomly divided into six groups using a strict random number table： blank group， model group， fluoxetine group （3.6 mg·kg^-1）， and high-， medium-， and low-dose Suanzaoren Tang groups （10， 5， 2.5 g·kg^-1），with 12 rats in each group. A depression model was established using isolation combined with CUMS. Fluoxetine and different doses of Suanzaoren Tang were administered continuously for 28 days. Behavioral indicators such as sucrose water consumption and open field test scores were recorded. Western blot and immunohistochemistry （IHC） were employed to analyze the expression of key proteins in the JNK/c-Myc/p53 signaling pathway， and the terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was used to evaluate the number of apoptotic cells in the hippocampus. ResultsCompared with the blank group， the model group exhibited a significantly reduced sucrose preference index （P<0.01）， a lower total score of horizontal and vertical movements in the open field test （P<0.01）， significantly increased expression of JNK， c-Myc， and p53 proteins in the hippocampus （P<0.01）， and a higher number of TUNEL-positive cells in the hippocampus （P<0.01）. Compared with the model group， the sucrose preference index and the total score of horizontal and vertical movements in the open field test significantly increased in the high- and medium-dose Suanzaoren Tang groups and the fluoxetine group （P<0.05， P<0.01）. The expression of JNK， c-Myc， and p53 proteins significantly decreased in all Suanzaoren Tang groups （high， medium， and low doses） and the fluoxetine group （P<0.05， P<0.01）. The number of TUNEL-positive cells in the hippocampus also significantly decreased in these groups （P<0.01）. ConclusionSuanzaoren Tang can regulate the expression of JNK/c-Myc/p53 proteins in the hippocampus of depression model rats， and its antidepressant mechanism may be related to its protective effect on hippocampal neurons.