Objective To investigate the expression and activation of MMP-2, MMP-9 in cerebral tissues of cerebral ischemia-reperfusion rats and the effects of doxycycline on the cerebral edema. Methods Cerebral ischemia-reperfusion model was established by middle cerebral artery occlusion (MCAO). The rats were randomly divided into a sham-operated and 6 reperfusion groups according to different reperfusion periods (3 h, 6 h, 12 h, 24 h, 72 h, and 120 h groups). Dry weight method was used to evaluate hemispheric water content of the ischemic side after treatment with doxycycline. Western blotting analysis and Gelatin zymography were used to determine the expression and activity of MMP-2 and MMP-9 in the ischemic cerebral tissues. Results The protein expression and activity of MMP-2 were significantly increased in the ischemia tissue at 3 h and 120 h after reperfusion(P<0.01) ; the protein expression and activity of MMP-9 began to increase at 6 h after reperfusion (P<0. 01) , reached the peak at 24 h, and then returned to the basal levels at 120 h after reperfusion (P>0.05). The hemispheric water content of the ischemic side gradually increased with reperfusion periods compared with the sham-operated group. Dexycycline significantly reduced the hemispheric water contents compared with the normal saline at the same time points(P<0.05 or P<0.01). Conclusion MMP-2, MMP-9 can induce vasogenic brain edema by degrading basal membrane of capillary vessel. The alteration of MMP-2, MMP-9 expression and activities are likely associated with brain edema during cerebral ischemia-reperfusion.

Objective: To explore the diagnostic value of fragmented QRS (fQRS) for coronary atherosclerotic heart disease (CHD), and to analyze it's relationship with left ventricular remodeling. Methods: From Nov. 2016 to Oct. 2018, 498 hospitalized patients in the Department of Cardiovascular Medicine of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine were selected consecutively. During the hospitalization, all the patients underwent coronary angiography. According to the angiographic results, the patients were divided into the control group (203 patients with negative or coronary stenosis < 30%), the mild to moderate stenosis group (155 patients with coronary stenosis 30% to 75%), and the severe stenosis group (140 patients with coronary stenosis≥75%). The incidences of fQRS(+) in the normal electrocardiogram among the three groups were compared by chi-square test of R×C contingency table. Two hundred and thirty patients with single-vessel stenosis≥30% were divided into the anterior descending branch group (128 cases), the right coronary branch group (59 cases), and the circumflex branch group (43 cases), and the relationship between fQRS(+) leads and diseased vessels was analyzed by nonparametric test. Finally, all the patients were divided into fQRS(+) group (86 cases) and fQRS(-) group (412 cases). The correlation between fQRS and left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), interventricular septum thickness (IVST) and left ventricular posterior wall thickness (LVPWT), respectively, were analyzed by binary Logistic regression model. Results: The chi-square test of R×C contingency table showed that the incidences of fQRS(+) in the three groups were 8.89%, 16.13% and 30.71%, respectively, with statistically significant differences (all P < 0.05). The nonparametric test showed that the fQRS(+) leads reflecting the anterior wall (V3, V4) were more common in the anterior descending branch group, and the fQRS(+) leads reflecting the interior wall and right ventricular (Ⅱ, III, AVF, V1, V2) were more common in the right coronary branch group, the fQRS(+) leads reflecting upper lateral wall (, AVL) were more common in the circumflex branch group, with statistically significant differences (all P<0.05). Binary Logistic regression analysis showed that fQRS was negatively correlated with LVEF (r=-0.030, OR=0.971, 95% CI 0.945-0.997, P=0.029), and positively correlated with LVESV (r=0.042, OR=1.043, 95% CI 1.005-1.082, P=0.026). Conclusion: fQRS has certain reference value in the clinical diagnosis of CHD, and left ventricular remodeling may be one of the mechanisms of fQRS.

Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.

OBJECTIVETo establish a method for organotypic slice culture of neonatal rat cortex in a modified condition and investigate the effect of spatial signals on neural stem cell (NSC) differentiation.

METHODSThe brain slices (200 µm in thickness) of neonatal SD rats (3 to 5 days old) were prepared and cultured in modified serum-free DMEM/F12 medium at 37 degrees celsius; with 95% O(2) and 5% CO(2). The organotypic slice cultures were observed regularly. NSCs isolated from the cortex of rat embryos (14-15 embryonic days) were cultured in serum-free DMEM/F12 supplemented with B27 and N2, and the passage 3 NSCs were labeled by CM-DiI before transplanted onto the organotypic slices cultured for 2 weeks. The survival of transplanted NSCs was assessed, and the cell differentiation was identified by immunofluorescence staining.

RESULTSThe organotypic slice cultures were well maintained for at least 4 weeks in the modified medium. The thickness of the organotypic slices reduced from 200 µm to 130 µm after 2-week culture in vitro due to the migration of the cells on the edge of the slices. CM-DiI-labeled NSCs survived well and differentiated into GFAP(+) glia and β-tubullin III(+) neurons.

CONCLUSIONNeonatal rat organotypic brain slice can be successfully cultured in a modified condition to serve as a model for studying NSC differentiation induced by spatial signals.

Animals ; Animals, Newborn ; Cell Differentiation ; physiology ; Cerebral Cortex ; cytology ; Coculture Techniques ; methods ; Fetus ; Neural Stem Cells ; cytology ; transplantation ; Organ Culture Techniques ; methods ; Rats

AIM To prepare chrysin solid lipid nanoparticles and to evaluate their pharmacokinetic behaviors.METHODS The particle size,Zeta potential and in vitro release rate of nanoparticles prepared by emulsification uhrasonication-low temperature solidification method were determined.Twelve SD rats were randomly divided into two groups and were intragastrically given suspensions of crude drug and nanoparticles,respectively.HPLC was used for the content determination of chrysin in plasma,after which blood drug concentration-time curves were drawn,and pharmacokinetic parameters were calculated.RESULTS The obtained nanoparticles demonstrated the particle size of (207.15 ±30.59) nm,PDI of (0.224 ±0.067) and Zeta potential of (-34.8 ±5.9) mV,respectively,whose accumulative release rate reached 84.36％ within 36 h.Their Cmax [(9.04 ± 1.52) μg/mL] and AUC0～t,[(33.67 ± 3.47) μg · h/mL] were much higher than those of the crude drug (P ＜ 0.01).CONCLUSION Solid lipid nanoparticles can promote the oral absorption and bioavailability of chrysin,with significant sustained-release characteristics.

BACKGROUNDOverexpression of Bcl-2 protein in cancer cells can inhibit programmed cell death and engender chemoresistance. Bcl-2 antisense oligonucleotide (G3139) has shown its antitumor effects enhanced in preclinical models when combined with taxol-based chemotherapy. This study aimed to investigate the efficacy of G3139 combined with epirubicin in the androgen-independent prostate cancer.

METHODSPC3 prostate cancer cell line was cultured and treated with epirubicin and Bcl-2 antisense oligonucleotide alone or in combination. The effects of therapeutic agents on cells were determined by the MTT assay. Expression of Bcl-2 mRNA and protein was documented by RT-PCR and Western blotting. Apoptosis induction was confirmed by flow cytometric analysis.

RESULTSBcl-2 antisense oligonucleotide alone produced no cytotoxic effects and the combination of Bcl-2 antisense oligonucleotide with epirubicin sensitized PC-3 cells to the killing effects of chemotherapy. A marked down-regulation of Bcl-2 mRNA and protein was observed after antisense and epirubicin cotreatment. A statistically significantly higher fraction of apoptotic cells was detected by flow-cytometric analysis after epirubicin treatment with prior antisense Bcl-2 transfenction, as compared with mono antisense Bcl-2 or epirubicin treatment.

CONCLUSIONThese data suggested that inhibition of Bcl-2 expression combined with epirubicin may be an attractive therapeutic strategy in hormone-refractory prostate cancer.

Apoptosis ; drug effects ; genetics ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; genetics ; Epirubicin ; pharmacology ; Flow Cytometry ; Humans ; Male ; Oligonucleotides, Antisense ; genetics ; Prostatic Neoplasms ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury.

METHODSPrimarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells.

RESULTSSalidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (P<0.05). Salidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (P<0.05).

CONCLUSIONSalidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.

Animals ; Caspase 3 ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Flow Cytometry ; Glucosides ; pharmacology ; Neural Stem Cells ; drug effects ; Phenols ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

The study was designed to explore the alteration of intracellular calcium concentration ([Ca²⁺]i), induced by transient receptor potential melastatin 8 (TRPM8) channel-specific agonist menthol, in pulmonary arterial smooth muscle cells (PASMCs) between control and pulmonary hypertensive (PH) rats. PH rat models were established by means of chronic hypoxia (CH) and monocrotaline (MCT) injection, respectively. PASMCs from control and PH rats were cultured. The change of [Ca²⁺]i in PASMCs induced by menthol, and the effect of TRPM8 channel-specific antagonist BCTC on the change of [Ca²⁺]i, were observed. Cellular localization of TRPM8 was examined by using immunohistochemistry. Results showed that menthol increased [Ca²⁺]i in the control PASMCs both in Ca²⁺ -normal and Ca²⁺ - free Tyrode's solutions, and at the same time BCTC could inhibit these two kinds of elevations. Compared with the control group, elevations of [Ca²⁺]i were decreased notably in CH- and MCT-pretreated PASMCs superfused with 2 mmol/L Ca²⁺ - or 0 Ca²⁺ -Tyrode's solutions. Immunohistochemical localization experiments showed that the whole PASMCs were dyed brown except for the nucleus. This study verified that TRPM8 exists both in membrane and sarcoplasmic reticulum of PASMCs. In addition, CH- and MCT-pretreatment could independently down-regulate the Ca²⁺ influx and Ca²⁺ release mediated by TRPM8 channel.
Animals ; Calcium ; metabolism ; Cells, Cultured ; Menthol ; pharmacology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Pulmonary Artery ; cytology ; Rats ; Sarcoplasmic Reticulum ; metabolism ; TRPM Cation Channels ; metabolism

OBJECTIVETo investigate the effect of baicalin on the behavioral characteristics of rats with attention deficit hyperactivity disorder (ADHD), and to provide a basis for further research on baicalin in the treatment of ADHD.

METHODSA total of 40 SHR rats were randomly divided into model group, methylphenidate hydrochloride (MPH) group, and low-, medium-, and high-dose baicalin groups, with 8 rats in each group. Eight WKY rats were selected as normal control group. The rats in the MPH group (0.07 mg/mL) and the low- (3.33 mg/mL), medium- (6.67 mg/mL), and high-dose (10 mg/mL) baicalin groups were given the corresponding drugs (1.5 mL/100 g) by gavage twice a day, and those in the normal control group and the model group were given an equal volume of normal saline by gavage twice a day. The course of treatment was 4 weeks for all groups. The open field test was performed to observe total moving distance and average moving speed on day 0 of experiment and at 7, 14, 21, and 28 days after gavage and to evaluate the control effects of drugs on hyperactivity and impulsive behavior. The Morris water maze test was used to observe the latency, time spent in the target quadrant, and number of platform crossings and to evaluate the effects of drugs on attention.

RESULTSThe open field test showed that the model group and the drug treatment groups had a significantly longer total moving distance and a significantly higher average moving speed than the normal control group on day 0 (P<0.05). On day 7, the MPH group had significant reductions in total moving distance and average moving speed compared with the model group (P<0.05). On day 14, the MPH group and the high-dose baicalin group had significant reductions in total moving distance and average moving speed compared with the model group (P<0.05). The data on days 21 and 28 showed that compared with the model group, the low-, medium-, and high-dose baicalin groups had gradual reductions in total moving distance and average moving speed (P<0.05). The water maze test showed that compared with the model group, the MPH group and the medium- and high-dose baicalin groups had a significantly longer time spent in the target quadrant (P<0.05), and the MPH group and the high-dose baicalin group had a significantly higher proportion of the moving distance in the target quadrant in total moving distance (P<0.05). The high-dose baicalin group had the highest number of platform crossings among all groups (P<0.05).

CONCLUSIONSBoth baicalin and MPH can regulate the motor ability and learning and memory abilities of SHR rats with ADHD and thus control the core symptoms of ADHD, i.e., hyperactivity, impulsive behavior, and inattention. Baicalin exerts its effect in a dose-dependent manner, and high-dose baicalin has the most significant effect, but compared with MPH, it needs a longer time to play its therapeutic effect.

Animals ; Attention Deficit Disorder with Hyperactivity ; drug therapy ; psychology ; Behavior, Animal ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; therapeutic use ; Male ; Maze Learning ; drug effects ; Motor Activity ; drug effects ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

OBJECTIVETo study the effect of baicalin on synaptosomal adenosine triphosphatase (ATPase) and lactate dehydrogenase (LDH) and its regulatory effect on the adenylate cyclase (AC)/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling pathway in rats with attention deficit hyperactivity disorder (ADHD).

METHODSA total of 40 SHR rats were randomly divided into five groups: ADHD model, methylphenidate hydrochloride treatment (0.07 mg/mL), and low-dose (3.33 mg/mL), medium-dose (6.67 mg/mL), and high-dose (10 mg/mL) baicalin treatment (n=8 each). Eight WKY rats were selected as normal control group. Percoll density gradient centrifugation was used to prepare brain synaptosomes and an electron microscope was used to observe their structure. Colorimetry was used to measure the activities of ATPase and LDH in synaptosomes. ELISA was used to measure the content of AC, cAMP, and PKA.

RESULTSCompared with the normal control group, the ADHD model group had a significant reduction in the ATPase activity, a significant increase in the LDH activity, and significant reductions in the content of AC, cAMP, and PKA (P<0.05). Compared with the ADHD model group, the methylphenidate hydrochloride group and the medium- and high-dose baicalin groups had a significant increase in the ATPase activity (P<0.05), a significant reduction in the LDH activity (P<0.05), and significant increases in the content of AC, cAMP, and PKA (P<0.05). Compared with the methylphenidate hydrochloride group, the high-dose baicalin group had significantly greater changes in these indices (P<0.05). Compared with the low-dose baicalin group, the high-dose baicalin group had a significant increase in the ATPase activity (P<0.05); the medium- and high-dose baicalin groups had a significant reduction in the LDH activity (P<0.05) and significant increases in the content of AC, cAMP, and PKA (P<0.05). Compared with the medium-dose baicalin group, the high-dose baicalin group had a significant increase in the ATPase activity (P<0.05).

CONCLUSIONSBoth methylphenidate hydrochloride and baicalin can improve synaptosomal ATPase and LDH activities in rats with ADHD. The effect of baicalin is dose-dependent, and high-dose baicalin has a significantly greater effect than methylphenidate hydrochloride. Baicalin exerts its therapeutic effect possibly by upregulating the AC/cAMP/PKA signaling pathway.

Adenosine Triphosphatases ; metabolism ; Adenylyl Cyclases ; physiology ; Animals ; Attention Deficit Disorder with Hyperactivity ; drug therapy ; physiopathology ; Cyclic AMP ; physiology ; Cyclic AMP-Dependent Protein Kinases ; physiology ; Flavonoids ; pharmacology ; therapeutic use ; L-Lactate Dehydrogenase ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Signal Transduction ; drug effects ; Synaptosomes ; chemistry ; ultrastructure