Objective To explore the role of hydrogen peroxide in mesenteric artery contraction of cirrhotic rats with portal hypertension,which was induced by bile duct ligation.Possible mechanism in RhoA/ROCK signal pathway was also part of the focus.Methods The bile duct ligation-induced cirrhotic rats and normal rats (control group) were treated equally with PEG-catalase(10 000 U/kg-1 · d-1,ip.) or by its vehicle for 8 days.Then the level of H2O2 in mesenteric arteries was detected.The contractile response to norepinephrine of arterioles was analyzed by vascular perfusion system.The protein expressions of the α1 adrenergic receptor,β-arrestin-2 and Rho kinase-1 (ROCK-1),and the activity of ROCK-1 were measured by western blot.In addition,the interaction of α1-adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation.Results Compared to normal rats,the dose-response curve of the mesenteric arterioles in response to norepinephrine shifted to the right,and the EC 50 increased in the rats with portal hypertension.PEG-catalase treatment can decrease the hydrogen peroxide level in arteries,thus significantly lowered EC50 and improved the reactivity to norepinephrine of the mesenteric arterioles in portal hypertension rats.No significant difference in the α1-adrenergic receptor amounts was observed among groups.There was remarkable decreases in the protein expressions of β-arrestin-2 and its interaction with the α1-adrenergic receptor in cirrhotic rats with PEG-catalase treatment.PEG-catalase also increased the amount and activity of ROCK-1 in cirrhotic rats.Conclusions The level of hydrogen peroxide increases in the mesenteric arteries in bile duct ligation-induced cirrhotic rats.And it enhances the β-arrestin-2 expression and its interaction with the α1-adrenergic receptor,which subsequently decreases the amount and activity of ROCK as well as the contractility of mesenteric arteries in response to vasoconstrictors.

Objective To investigate the predictive value of left ventricular diastolic function on mechanical ventilation weaning in patients with left ventricular ejection fraction (LVEF) > 0.50.Methods A retrospective case control study was conducted. Sixty-five patients with LVEF > 0.50 undergoing mechanical ventilation for more than 48 hours admitted to surgery intensive care unit (ICU) of China-Japan Friendship Hospital from June 2014 to December 2016 were enrolled. The clinical data and parameters of echocardiography before spontaneous breathing trial (SBT) were collected. The possible relationship between left ventricular diastolic function and the results of weaning was analyzed according to analysis of blood flow filling parameters of mitral valve orifice. According to the grading standard of left ventricular diastolic function, the patients were divided into normal, mild (level 1) and moderate to severe (level 2-3) groups, and the outcomes of weaning were compared among the groups. Then patients were also divided into two groups of weaning successfully and weaning failure, and the clinical data and left ventricular diastolic function parameters of patients were compared between the two groups. The predictive value of left ventricular diastolic function on results of weaning was evaluated with receiver operating characteristic curve (ROC).Results Sixty-five patients were enrolled and 28 patients (43.1%) failed weaning, 22 patients failed the first SBT and 6 required reintubation within 48 hours, 31 of the patients presented normal left ventricular diastolic function, 9 of patients presented mild diastolic dysfunction, and 25 of them presented moderate to severe diastolic dysfunction. So with the gradual increase of the severity of diastolic dysfunction, the rate of weaning failure was gradually increased, which was 16.1%, 44.6% and 76.0% respectively (χ2 = 20.240,P = 0.001). Patients who failed weaning presented evidence of increased left ventricular filling pressures at pre-SBT, by demonstrating decreased deceleration time of E (DTE, s: 180.4±5.1vs. 196.8±4.0,t = 2.567,P = 0.013), increased left ventricular mitral valve diastolic early and late filling velocity ratio (E/A: 1.47±0.08 vs. 1.14±0.05, t = 3.827,P = 0.000), increased lateral, septal and averaged left ventricular mitral valve diastolic early velocity and maximal velocity of mitral annulus in early diastolic velocity ratio (E/Em: 10.26±0.52 vs. 7.28±0.41,t = 4.535, P = 0.000; 10.08±0.58 vs. 8.16±0.40,t = 2.797,P = 0.007; 10.17±0.48比7.72±0.35,t = 4.231,P = 0.000), and the rapid shallow breathing index (RSBI) was also increased significantly (61.7±3.6 vs. 50.6±2.7,t = 2.507, P = 0.015). It was shown by ROC curve analysis that the basic left ventricular diastolic function at pre-SBT had the diagnostic performance in predicting the outcome of weaning from mechanical ventilation, especially E/A and lateral E/Em. Pre-SBT values of E/A greater than 1.2 and lateral E/Em greater than 7.9 predicted weaning failure with an area under the ROC curve (AUC), sensitivity, and specificity of 0.81±0.06 and 0.85±0.06, 82.6% and 91.3%, 81.4% and 80.7%, respectively, and the AUC was higher than RSBI (0.70±0.07). The AUC of combination of E/A > 1.2 and lateral E/Em > 7.9predicting weaning failure was 0.86±0.05 with a sensitivity of 78.3% and a specificity of 93.6%.Conclusions The results suggest that left ventricular diastolic dysfunction is significantly associated with weaning outcome in critical patients with LVEF > 0.50. The combination of E/A ratio greater than 1.2 and E/Em ratio greater than 7.9 may identify patients at high risk of weaning failure.

AIMTo observe the effects of lidocaine and thiopental on the neuronal injury induced by the experimental ischemia in hippocampus slice cultures obtained from postnatal 22 days SD rats.

METHODSModel of the experimental ischemia was produced by hypoxia and glucose deprivation. Propidium iodide (PI) assay was used to observe the neuronal injury in CA1 and dentate gyrus (DG).

RESULTSAfter experimental ischemia, the peak of PI index was appeared in CA1 and DG on the first day (P < 0.01), PI index in DG was less than in CA1 (P < 0.01). PI indices were still higher during seven days after the experimental ischemia than before the experimental ischemia (P < 0.01). 10 nmol/L and 100 nmol/L concentration of lidocaine could significantly decrease PI indices in CA1 and DG (P < 0.01). 250 nmol/L and 600 nmol/L concentration of thiopental also decreased the PI indices in CA1 and DG (P < 0.01). The neuronal injury peaks were postponed to the third day after the experimental ischemia by lidocaine and thiopental.

CONCLUSIONIt suggested that lidocaine and thiopental could decrease the neuronal injury in CA1 and DG induced by the experimental ischemia, and postpone the neuronal injury peaks to the third day after the experimental ischemia.

Animals ; Brain Ischemia ; pathology ; CA1 Region, Hippocampal ; drug effects ; pathology ; In Vitro Techniques ; Lidocaine ; pharmacology ; Neurons ; drug effects ; pathology ; Rats ; Thiopental ; pharmacology

Chronic kidney disease (CKD) is a serious chronic disease with high incidence, poor prognosis, and a variety of complications. Indoxyl-sulfate (IS) and p-cresol sulfate (PCS) are two typical gut-derived uremic toxins, which are produced by the co-metabolism of intestinal microbes and the host. With the progression of CKD, gut-derived uremic toxins such as IS and PCS accumulate in patients with CKD and thereafter accelerate the progression of CKD. Gut microbiota is closely related with CKD, and targeting gut microbiota to regulate gut-derived uremic toxins synthesis and metabolic pathways may be a promising strategy to delay the progression of CKD. In this paper, the relationship between gut microbiota, gut-derived uremic toxins, and CKD was analyzed, and the strategy to delay the progression of CKD by targeting gut microbiota and uremic toxins metabolism pathway was proposed.

Objective To investigate the distribution of death among human immunodeficiency virus/acquired immuno deficiency syndrome(HIV/AIDS) cases in Guizhou Province from 1995 to 2017. Methods The HIV/AIDS death cases from 1995 to 2017 were downloaded from “Chinese National Comprehensive HIV/AIDS Prevention and care Information system” in Guizhou Province and were analyzed. Results From 1995 to 2017, Guizhou Province reported a total of 43 794 HIV/AIDS cases and 11 527 deaths according to current address. After excluding missing persons, the HIV/AIDS mortality rate of the province was 29.8%. The proportion of reported HIV/AIDS cases died in the same year ( 21995-2012=139.5, P<0.001; 22012-2015=28.2, P<0.001) and the proportion of HIV/AIDS cases ( 21995-2012=109.1, P<0.001; 22012-2014=57.2, P<0.001) who survived at the beginning but died later in the year all showed a trend being low-high-low. In the analysis of the detection history of death cases, the detection proportion of cluster of differentiation 4(CD4) T-cell and the proportion of antiviral treatment had been increasing year by year. The analysis of the cause of death found that the proportion of death caused by AIDS increased firstly and then declined, and the proportion of death due to excessive drug abuse showed a trend of declining year by year. Conclusions The mortality rate of HIV/AIDS in Guizhou Province was still high, and decreased rather slow. Expanding the coverage of HIV monitoring and screening is one of the key tasks of AIDS prevention and control. CD4+T-cell testing and free antiviral treatment should be strengthened to reduce the mortality rate of HIV/AIDS in Guizhou Province in the future.

Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

AIMTo investigate the effect of diazoxide preconditioning and the role of ERK and JNK in cellular signaling during diazoxide preconditioning protection in isolated spontaneous hypertension rat (SHR) hearts.

METHODSHearts were isolated from male SHR rats, and perfused on a Langendorff apparatus. Five groups were considered (n = 6). Con: after 40 min perfusion the hearts were submitted to 25 min ischemia followed by 30 min reperfusion. IP: the hearts were preconditioned with 2 periods of 5 min ischemia and 10 min reperfusion prior to 25 min ischemia. DP: the hearts were preconditioned with 2 periods of 10 min K-H solution with 50 micromol x L(-1) diazoxide and 5 min K-H solution reperfusion prior to 25 min ischemia. 5-HD: perfuse with 100 micromol x L(-1) 5-HD (a special mitochondrial ATP sensitive potassium channel blocker) for 10 min followed by 30 min K-H solution perfusion before 25 min ischemia. 5-HD + DP: 100 micromol x L(-1) 5-HD was given for 10 min before diazoxide preconditioning.

RESULTSDuring reperfusion, comparing with Con group, the recoveries of left ventricle developed pressure (LVDP), + dP/dt(max), - dP/dt(max) and left ventricle end diastolic pressure (LVEDP) were improved in IP and DP groups (P < 0.01 vs Con). At the end of reperfusion, compared with Con group, the expression of ERK in myocardium were higher in IP and DP groups (P < 0.01 vs Con), there was no significance between 5-HD and Con group, but 5-HD couldn't inhibit the expression of ERK induced by diazoxide preconditioning. The expression of JNK in IP and DP groups were decreased (P < 0.05 vs Con), this effect could been inhibited by 5-HD.

CONCLUSIONThese results indicated that diazoxide preconditioning could mimic ischemic preconditioning, the activation of ERK expression and the declining of JNK expression involved in diazoxide preconditioning in isolated SHR hearts.

Animals ; Diazoxide ; pharmacology ; In Vitro Techniques ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Signaling System ; Male ; Myocardial Ischemia ; metabolism ; physiopathology ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Myocardium ; metabolism ; Rats ; Rats, Inbred SHR

OBJECTIVETo prepare highly specific anti-ouabain polyclonal antibody for detecting endogenous ouabain in tissues.

METHODSOuabain-BSA compound was used to immunize hens, and the eggs were collected one week after the first immunization. The IgY antibodies in the egg yolk were separated and purified by PEG-6000 Method, and analyzed by 12% SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) for titration. The IgY antibodies obtained were applied subsequently in ELISA and immunohistochemistry.

RESULTSThe IgY titer increased rapidly after the second immunization, with the highest titer of 1:10240 that lasted for at least 4 weeks. Competitive ELISA for IgY detection showed an average intraassay coefficient of variation (CV) of 2.03% and an inter-assay CV of 2.34%. Immunohistochemistry visualized the location of the endogenous ouabain mainly in the cytoplasm of the zona reticularis of rat adrenal cortex.

CONCLUSIONImmunization of hens allows efficient preparation of IgY antibody which can be used in routine immunoassays.

Animals ; Calibration ; Cattle ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Immunization ; methods ; Immunoglobulins ; immunology ; Immunohistochemistry ; Ouabain ; analysis ; immunology ; Rats

In formalin pain model, the effect of propofol on Fos expression in the spinal cord was examined by means of c -fos oncogene immunohistochemistry and NADPH-d histochemistry. Fos-like immunoreactive (FLI) neurons were mainly found in the ipsilateral dorsal horn after injection of formalin, some of which were FLI/NOS double-labeled neurons. Most of the FLI or FLI/NOS double-labeled neurons were observed in the medial part of lamina and the outer lamina . Before or after injection of formain, i.p. injection of propofol significantly decreased the number of FLI and FLI/NOS double-labeled neurons in all laminae (P<0.05 or P<0.01). By single i.p. injection of propofol or normal saline, few FLI neurons were found in the spinal cord. The results suggest that the antinociceptive function of propofol is possibly involved in the depression of the NOS neurons in the spinal cord.
Animals ; Female ; Formaldehyde ; Male ; Neurons ; metabolism ; Nitric Oxide Synthase ; metabolism ; Pain ; chemically induced ; metabolism ; Propofol ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism

OBJECTIVETo evaluate the outcome of the treatment of malocclusion with mandibular congenital missing teeth guided by Bolton index.

METHODS26 class I and II cases (males 10, females 16) were chosen in this study. Guided by Bolton index, enamel stripping, denture and two types of tooth extraction were used in the treatment.

RESULTSNormal Bolton index was achieved and the results of treatment were satisfactory.

CONCLUSIONSMalocclusion with mandibular congenital missing teeth can be treated successfully guided by Bolton index.

Adolescent ; Adult ; Anodontia ; complications ; Cephalometry ; methods ; standards ; Child ; Female ; Humans ; Male ; Malocclusion ; etiology ; therapy ; Orthodontics, Corrective ; methods ; standards ; Treatment Outcome