Analysis of a Pedigree with Dopa-responsive Dystonia Caused by Novel Mutation of TH Gene / 中山大学学报(医学科学版)

@#【Objective】 To study the mutation characteristics of Tyrosine hydroxyls（TH） gene in a pedigree with dopa-responsive dystonia（DRD）. 【Methods】 Extraction of genomic DNA from peripheral blood of a proband and his parents and two sisters using high- throughput sequencing （NGS） method were detected on 256 known pathogenicity genes associated with dystonia and dyskinesia.【Results】Mutations on tyrosine hydroxylase（TH）gene in the exon 14 and exon 9 were detected in the proband and his eldest sister in this pedigree. They had a complex heterozygosity of c.1481C > T（p.Thr494Met）and c.943G >A（p.Gly315Ser），and one heterozygous mutation was carried by parents respectively. The mutation was not detected in his second sister and 50 people with normal phenotype controls. 【Conclusion】 The mutations of TH gene c. 1481C > T（p.Thr494Met）and c. 943G > A（p.Gly315Ser）led to the gene abnormality in DRD family，and a new mutation of TH gene was found，which expanded the relationship between DRD genotype and clinical phenotype. It is vital that early accurate diagnosis and treatment of DRD is the key to improve prognosis.

Prevalence of urogenital tract infections with Ureaplasma urealyticum or HPV among men of reproductive age and the associated factors / 中华男科学杂志

Objective@#To investigate the prevalence of urogenital tract infections with Ureaplasma urealyticum (UU) and human papilloma virus (HPV) in males of reproductive age and the associated factors.@*METHODS@#Using the multi-stage cluster sampling method and a structured questionnaire, we conducted an investigation among 18－50 years old males in Songjiang District, Shanghai, from August 2016 to July 2018. We collected secretory specimens from the urogenital tract of the subjects and detected the infections of UU and HPV by laboratory examination.@*RESULTS@#Among the 621 males included in this study, 279 (44.93%) were found infected with UU, 18 (2.90%) with HPV, and 15 (2.42%) with both UU and HPV. Univariate analysis showed that smokers had a higher rate of UU infection (50.54% ［140/277］) than non-smokers (40.41 ［139/344］), and those with senior high school or secondary technical school education had a higher rate of HPV infection (4.84% ［12/248］) than others (1.61% ［6/373］). Binary stepwise logistic regression analysis revealed a higher risk of UU infection in the subjects with junior high school or lower education than in others (OR = 0.61, 95% CI: 0.39－0.96) as well as in smokers than in non-smokers (OR = 1.46, 95% CI: 1.01－2.01).@*CONCLUSIONS@#The prevalence of UU infection is high, while that of HPV is low among men of reproductive age in Songjiang, Shanghai. The screening of UU infection should be enhanced among men of reproductive age, especially among smokers and those with lower education.

Infiltration of tumor associated macrophages in multiple myeloma and its clinical significance / 中华血液学杂志

Objective: To investigate the clinical significance of tumor associated macrophages (TAM) in multiple myeloma (MM) and the relationship with angiogenesis and immunosuppression. Methods: Seventy cases of MM patients diagnosed from August 2015 to June 2017 were enrolled in the study as experimental group, 20 cases of benign hematological diseases (13 with iron deficiency anemia and 7 with megaloblastic anemia) patients as control group. Immunohistochemical method was used to detect the expression of CD163, CD34 and VEGF in bone marrow samples, and flow cytometry was used to detect the proportion of regulatory T cell (Treg cells), ELISA was used to detect the level of IL-10, and the clinical features were analyzed. Results: ①Among the 70 patients, there were 31 males and 39 females with a median age of 65 (50~78) years old. TAM infiltration density, microvascular density (MVD), VEGF expression level, Treg ratio and IL-10 level in bone marrow samples of 70 MM patients were significantly higher than those of benign hematological diseases (P<0.05). ②In the MM group, the above indexes of the patients with disease stabilized (15 cases) were lower than those of the newly diagnosed group (35 cases) and the relapse refractory group (20 cases) (P<0.05), those of relapse refractory group were higher than those of newly diagnosed group (P>0.05). ③Of the 35 newly diagnosed MM patients, 27 completed 4 courses of treatment. In the effective group (15 cases), the TAM infiltration density after treatment was significantly lower than that before treatment, the difference was statistically significant[(20.20±7.66) vs (28.87±11.97), t=2.362, P=0.025]; while in the ineffective group of 12 cases, the difference of the TAM infiltration density before and after treatment was not statistically significant[(42.00±13.76) vs (48.25±13.59), t=1.119, P=0.275]. ④TAM infiltration density in the effective group after bortezomib treatment (21 cases) were lower than those in the non-bortezomib treatment group (18 cases)[(16.52 ±4.26) vs (19.27 ±5.82), t=1.662, P=0.170]. ⑤The TAM infiltration density in MM patients was positively correlated with MVD, VEGF expression level, Treg cell ratio and IL-10 level (P<0.001). Conclusion: The infiltration of TAM in the microenvironment of MM, which may promoting angiogenesis and inhibiting immune response, is related to the occurrence, development, therapeutic effect and drug resistance of MM.

Safety evaluation of Parecoxib Sodium for Injection / 药物评价研究

Objective To evaluate the safety of Parecoxib Sodium for Injection.Methods Systemic active anaphylaxis test:Guinea pigs were injected respectively with Parecoxib Sodium for Injection (test sample,20 mg/mL),primary control,sodium chloride injection or human serum albumin,once every other day,continuously for three times.After 14 and 21 d from the end time of sensitization,to stimulation and to observe whether allergic reactions occurred within 30 mins.Passive cutaneous anaphylaxis test:Guinea pigs were received 0.1 mL antiserum injection for inducing passive sensitization,after 24 h of that we stimulated the guinea pigs,and the guinea pigs were sacrificed after 30 mins to examine the diameter of blue spots.Blood vessel irritation test:After continuous ear Ⅳ test sample for 5 d,HE staining was performed at the end of the administration and recovery period,and the stimulation of the blood vessel at the site of injection was observed.Hemolysis or agglutination of Parecoxib Sodium for Injection was examined by in vitro methods.Results Under the dosage of 20 mg/mL,guinea pigs showed no systemic allergy and passive skin allergy,and no hemolysis,agglutination,and irritation of vascular was observed.Conclusion Under the present experimental conditions,20 mg/mL Parecoxib Sodium for Injection shows no obvious allergic reactions,irritation and hemolysis,is safe.

Safety evaluation of Parecoxib Sodium for Injection / 药物评价研究

Objective To evaluate the safety of Parecoxib Sodium for Injection.Methods Systemic active anaphylaxis test:Guinea pigs were injected respectively with Parecoxib Sodium for Injection (test sample,20 mg/mL),primary control,sodium chloride injection or human serum albumin,once every other day,continuously for three times.After 14 and 21 d from the end time of sensitization,to stimulation and to observe whether allergic reactions occurred within 30 mins.Passive cutaneous anaphylaxis test:Guinea pigs were received 0.1 mL antiserum injection for inducing passive sensitization,after 24 h of that we stimulated the guinea pigs,and the guinea pigs were sacrificed after 30 mins to examine the diameter of blue spots.Blood vessel irritation test:After continuous ear Ⅳ test sample for 5 d,HE staining was performed at the end of the administration and recovery period,and the stimulation of the blood vessel at the site of injection was observed.Hemolysis or agglutination of Parecoxib Sodium for Injection was examined by in vitro methods.Results Under the dosage of 20 mg/mL,guinea pigs showed no systemic allergy and passive skin allergy,and no hemolysis,agglutination,and irritation of vascular was observed.Conclusion Under the present experimental conditions,20 mg/mL Parecoxib Sodium for Injection shows no obvious allergic reactions,irritation and hemolysis,is safe.

Correlation of the Desialylation of Platelets with Efficacy of the First-line Therapy for ITP / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect desialylation of platelets in primary immune thrombocytopenia(ITP) patients with FITC-labelled ECL and RCA-1, and compare the correlation of the desialylation level and the efficacy of first-line therapy for ITP.</p><p><b>METHODS</b>Before treatment, 48 ITP patients were selected and their levels of ECL and RCA-1 were detected with flow cytometry.</p><p><b>RESULTS</b>The desialylation level in the different efficacy groups by using the first-line therapy of corticosteroids and (or) intravenous immunoglobulin G (IVIG) had a statistically significant difference (P<0.05). The correlation analysis showed negative relation of the therapeutic efficacy with desialylation level, that is to say, the more high of desialylation level, the more poor therapeutic efficacy of the first-line therapy.</p><p><b>CONCLUSION</b>The desialylation level of platelets in ITP patients is related with the first-line therapeutic efficacy, the efficacy for patients with high desialylation level is poor, suggesting that the FcR-independent pathway exists in clearance of platelets in ITP patients. Therefore, the desialylation level of platelets may suggest the first-line therapeutic efficacy for ITP patients to a certain degree, and may be used as a potential target for the treatment of refractory ITP.</p>

The First Switched Time of PML/RARα Fusion Gene in Patients with Acute Promyelocytic Leukemia and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the first switched time of PML/RARα fusion gene in patients with acute promyelocytic leukemia (APL) and its clinical significance.</p><p><b>METHODS</b>sixty cases of newly diagnosed APL were enrolled in this study. They received standard remission induction, consolidation and maintenance treatments according to the clinical pathway for APL, and were followed up. During the same time the PML/RARα fusion gene mRNA expression of all cases was detected by multi-nested PCR.</p><p><b>RESULTS</b>except for 3 death cases and 1 case failed to follow-up, the PML/RARα fusion genes in the remaining 56 cases were firstly found to be negative from 24 to 381 days respectively, the mean value of the first switched time was 131 ± 90 days. There was no statistically significant difference in age, sex and risk stratification between different groups. However, the cases with L-type PML/RARα gene had shorter time compared with the patients with S-type PML/RARα gene (P = 0.032); then, for the above-mentimed 56 cases, the follow-up duration ranged from 25-1979 days (median 946 days), long-term molecular remissions had been observed in most cases, but 1 case with the first switched time of 133 days unfortunately recurred to be positive and followed by clinical relapse.</p><p><b>CONCLUSION</b>The PML/RARα fusion gene in newly diagnosed APL patients was first switched to be negative in about 4 months after treatment. The first switched time of PML/RARα fusion gene can objectively reflect the reduction of leukemia cells, and the differences among different subtypes of PML/RARα fusion gene may have some suggestions for the treatment, but without important significance for the evaluation of prognosis and recurrence for APL patients. In addition, minimal residual disease (MRD) can be dynamically monitored by detecting PML/RARα fusion gene, thus having an important clinical significance for analysis of APL recurrence.</p>

Effects of total flavones of Elsholtzia splendens in isolated ischemia/reperfusion rat hearts / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the influence of total flavonoids of Elsholtzia splendens (TFES) on isolated ischemia/reperfusion rat hearts and its underlying mechanisms.</p><p><b>METHODS</b>Hearts isolated from male SD rats were perfused on the Langendorff apparatus and subjected to global ischemia for 30 min followed by 120 min of reperfusion. The cardiac infarct size was measured by TTC staining. Hemodynamic parameters and the level of lactate dehydrogenase (LDH) in the coronary effluent were measured. Absorbance at 520 nm was determined in isolated cardiac mitochondria exposed to 200 micromol/L CaCl2 to detect the opening of the mitochondrial permeability transition pore.</p><p><b>RESULTS</b>Pretreatment with TFES (1, 10, 100 microg/ml) for 5 min decreased infarct size and LDH release and improved the recovery of the left ventricular developed pressure. In mitochondria, the decrease of absorbance at 520 nm evoked by CaCl2 was greatly inhibited by TFES.</p><p><b>CONCLUSION</b>TFES prevents myocardial ischemia/reperfusion injury, and this cardioprotective effect is probably via inhibiting mitochondrial permeability transition pore opening.</p>

The effect of chlamydiaphage phiCPG1 capsid protein Vp1 on the Chlamydia trachomatis / 中华微生物学和免疫学杂志

Objective To investigate the effect of recombinant chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia trachomatis(Ct) after Vp1 was co-cultured with Ct (reference strains and clinical strains).Methods The recombinant chlamydiaphage phiCPG1 capsid protein Vp1 was expressed and purified.Equal amount of Ct standard strains (E/UW-5/Cx and D/UW-3/Cx) or clinical strains,which had been incubated with Vp1 protein at the concentration of 53 μg/ml for 3 h at room temperature,were inoculated into McCoy.After cell culture,idione stain and transmission electron microscope were used to observe the effect of Vp1 on the Ct.The effect of Vp1 protein on the cell line McCoy was determined by MTT assay,the responses of Escherichia coli BL21 and DH5α toward Vp1 protein were determined using broth microdilution assays.Results Vp1 had obviously inhibitive effect on Ct,the inhibition ratios were about 40％-70％in clinical strains,72％ in reference strain D and 78％ in E,respectively.Abnormally enlarged RBs were observed after Vp1-treatment and Vp1 could arrest chlamydial developmental cycle using electron microscope.There was no effect of Vp1 on McCoy cells or bacteria BL21 or DH5α.Conclusion The recombinant Vp1 from phiCPG1 has obviously inhibitive effect on the growth of Ct,it will be helpful for the treatment of Ct infection in clinic.

Nuclear matrix proteins differentially expressed in human prostate cancer cell lines and benign prostatic hyperplasia epithelial cell line / 中华男科学杂志

<p><b>OBJECTIVE</b>To compare the expression of nuclear matrix proteins (NMPs) in benign prostatic hyperplasia (BPH) epithelial cell line BPH-1 versus those in androgen-dependent human prostate cancer cell line LNCap and androgen-independent prostate cancer cell line PC-3.</p><p><b>METHODS</b>We isolated NMPs from the BPH-1, LNCap and PC-3 cell lines by 2-dimensional electrophoresis (2-DE), analyzed the differentially expressed proteins by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS), and identified them by peptide mass fingerprint and database searching.</p><p><b>RESULTS</b>We successfully obtained well-resolved reproducible 2-DE patterns of NMPs in human prostate cancer cell lines, identified 12 differentially expressed NMPs including enzymes, regulatory proteins, RNA-binding protein and various other factors, 3 up-regulated and 9 down-regulated in prostate cancer cell lines.</p><p><b>CONCLUSION</b>There are obvious differences in the expressions of NMPs between human prostate cancer cell lines and benign prostatic hyperplasia epithelial cell line.</p>

Effect of S-allyl-L-cysteine on isolate heart subject to ischemia/reperfusion / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of S-allyl-L-cysteine (SAC) on isolated rat heart subject to ischemia/reperfusion(I/R) injury and the mechanisms.</p><p><b>METHODS</b>The isolated perfused rat hearts on a Langendorff apparatus were subjected to global ischemia for 30 min and followed by 120 min of reperfusion. Hemodynamic index, the production of formazan and the level of lactate dehydrogenase (LDH) in the coronary effluent were determined. Superoxide dismutase (SOD) and reactive oxygen species (ROS) in myocardial homogenates were measured.</p><p><b>RESULTS</b>Compared with I/R group, the hemodynamics were greatly improved, the production of formazan was increased, and LDH level in effluent was reduced in SAC group. SAC improved the SOD activity and significantly decreased the level of ROS. In addition, threonine (Thr) attenuated the protective effect of SAC significantly.</p><p><b>CONCLUSION</b>SAC has protective effect against myocardial ischemia/reperfusion injury on rats. The possible mechanism is that SAC be transported into the cell through alanine-serine-cysteine-transporter 1 (ASCT-1) improves SOD activity and reduces the level of ROS.</p>

Enhance efficiency of adenovirus on AAV transfering human RPE cells / 中华实验眼科杂志

Background Adeno-associated virus-based vector is one of most efficient vehicles.It presents with a long-term and efficient transfer and expression of therapeutic genes with minimal toxicity.But its delayed-and low-efficient transgene expression limits the application of AAV vector.To explore an improving method of AAV infecting RPE cells is the hot spot. Objeetive Present study was to investigate whether adeno-associated virus (AAV)combined with low dose non-replieable adenovirus(Ad-null)can enhance its infection efficieney on RPE ceils in vitro. Methods Human RPE cells were isolated from the donate eyeballs under the approval of the Ethic Committee of this hospital.The cells were cultured in DMEM containing 10%fetal bovine serum.AAV particles with enhanced green fluorescence protein(EGFP)were added into the medium alone or in combination with different amount of adenovirus for 30 days.The cells were detected under the fluorescence microscope,and the protein expression levels of report gene EGFP in RPE cells were analyzed with Western blotting assay. Results Melanin granules could be found in cultured RPE cells.EGFP was expressed in RPE cells at 2 days after AAV-EGFP infection and peaked at 12 days and remained for about 3-week duration,showing the green influorescence under the influorescence mwroscope.After the cells were infected by AAV2-EGFP with 0.01 to 1000 MOI Ad-null respectively,the number of cells with green influorescence was obviously increased with the enhanced infiuorescence intensity.The enhance of the infection efficiency began in the 0.1 MOI Ad-null group and peaked in 10 MOI Ad-null group.Dead cells were exhibited in the 100 or more MOI Ad-nulor group.Western blotting assay demonstrated that the protein expression level of EGFP in RPE cells enhanced significantly in 1 and 10 MOI Ad-null groups compared with only AAV infection group. Conclusion These finding suggested that the infection efficiency of AAV can be improved significantly when it is used with low dose Ad-null in vitro.This offers a basis for further study of gene therapy.

Luteolin reduces cardiac dysfunctions in streptozotocin-induced diabetic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of luteolin (Chinese Traditional Medicine) on cardiac functions and mitochondrial oxidative stress in streptozotocin (STZ)-induced diabetic rats.</p><p><b>METHODS</b>Male SD rats were randomly divided into a normal control group, a luteolin control group, a diabetic group, and diabetic groups orally administered with a low dose (10 mg/(kg x d)) or a high dose of luteolin (100 mg/ (kg x d)) for eight weeks. The body weight, blood glucose, cardiac functions, left ventricular weight, myocardial collagen and reactive oxygen species (ROS) levels were assayed. The cardiac mitochondrial ROS level, superoxide dismutase (SOD) activity and the mitochondrial swelling were measured.</p><p><b>RESULTS</b>Treatment with luteolin had no effect on the blood glucose but reduced the losing of body weight in diabetic rats. High dose of luteolin markedly reduced the ratio of ventricular weight and body weight, increased the left ventricular develop pressure, and decreased the left ventricular end diastolic pressure in diabetic rats. The myocardial levels of ROS and collagen, the cardiac mitochondrial ROS level, and the mitochondrial swelling in diabetic rats were all markedly reduced by high dose of luteolin. Furthermore, high dose of luteolin significantly increased the mitochondrial SOD activity in diabetic rat hearts.</p><p><b>CONCLUSION</b>Treatment with luteolin for 8 weeks markedly improves the cardiac function, which may be related to reducing mitochondrial oxidative stress and mitochondrial swelling in diabetic rats.</p>

Auricularia auricular polysaccharide protects myocardium against ischemia/reperfusion injury / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To determine whether auricularia auricular polysaccharide (AAP) protects heart against ischemia/reperfusion (1/ R) injury and its underlying mechanisms.</p><p><b>METHODS</b>Male Sprague-Dawley rats, pretreated with AAP (50, 100, 200 mg/(kg x d), gastric perfusion) for 4 weeks, were used for Langendorff isolated heart perfusion. The hearts were subjected to global ischemia for 30 min followed by 120 min of reperfusion and the left ventricular hemodynamic parameters were measured. Formazan, a product of 2, 3, 5-triphenyl-tetrazolium chloride (TTC), which is proportional to myocardial viability, was measured at 490 nm, and the level of lactate dehydrogenase (LDH) in the coronary effluent was measured to evaluate the cardiac injury. The cardiac malondialdehyde (MDA), a product of lipid peroxidation, and superoxide dismutase (SOD) activity were determined after myocardial I/R.</p><p><b>RESULTS</b>The pretreatment with AAP at 50, 100, 200/(kg d) for 4 weeks before I/R increased myocardial formazan content, reduced LDH release, improved the recovery of the left ventficular developed pressure, maximal rise rate of left ventricular pressure, and rate pressure product (left ventricular developed pressure multiplied by heart rate) attenuated the decrease of coronary flow during reperfusion. The cardiac protective effect of high dose AAP was more potent than that of compound radix salviae miltiorrhizae (CRSM, 4 ml/(kg x d), gastric perfusion for 4 weeks). Pretreatment with AAP (100 mg/(kg x d)) markedly inhibited the increase of MDA level and the decrease of SOD activity induced by I/R in myocardium.</p><p><b>CONCLUSION</b>The findings indicate that in the isolated rat heart, AAP protects myocardium against ischemia/reperfusion injury via enhancing the activity of SOD and reducing lipid peroxidation in heart.</p>

The study of effects and mechanism of U50, 488H on electrical coupling during ischemia in the perfused isolated rat heart / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To determine the effect of activation of lambda-opioid receptor with U50, 488H, a selective kappa-opioid receptor agonist, on the changes in electrical coupling during prolonged ischemia and to explore the possible mechanism.</p><p><b>METHODS</b>The isolated rat heart was perfused in a Langendorff apparatus. The effect of U50, 488H on electrical coupling parameters including onset of uncoupling, plateau time, slope and fold increase in r(t) was observed in isolated perfused rat heart subjected to global no-flow ischemia. The effect of U50, 488H on connexin 43 (Cx43) expression of ventricular muscle during ischemia was determined by immunohistochemistry.</p><p><b>RESULTS</b>In the prolonged ischemia model, U50, 488H concentration dependently delayed the onset of uncoupling, increased time to plateau, and decreased the maximal rate of uncoupling during ischemia. The effect of U50, 488H on electrical uncoupling parameters during ischemia was abolished by a selective kappa-opioid receptor antagonist nor-BNI or a PKC inhibitor chelerythrine. The amount of Cx43 immunoreactive signal in ventricular muscle was greatly reduced after ischemia. U50, 488H markedly increased Cx43 expression during ischemia and its effect was also attenuated by nor-BNI or chelerythrine.</p><p><b>CONCLUSION</b>These results demonstrated that U50, 488H delayed the onset of uncoupling and plateau time, decreased the maximal rate of uncoupling and increased Cx43 expression of ventricular muscle during ischemia, and these effects of U50, 488H were mediated by kappa-opioid receptor, in which activation of PKC was involved. The effect of U50, 488H on electrical coupling during ischemia was probably correlated with preservation of Cx43 in cardiac muscle.</p>

Myocardial protective effect of acetylcholine against ischemia/reperfusion injury and its underlying mechanism / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To determine whether the caidioprotection of acetylcholine (ACh) against ischeniia/reperftision (I/R) injury is re-kited to mitochondrial permeability transition pore (MEW) and mitochondrial AW-sensitive potassium channel (mitoK(ATP)).</p><p><b>METHODS</b>Male Sprague-Dawley rats were used for Langendorif isolated bean perkision. The hearts were subjected to global ischemia for 30 mm followed by 120 rein of reperfusion and the left ventricular hemodynaniic parameters were measured. Formazan, a product of 2,3, 5-triphenyl-tetrazolium chloride (TTC), which is proportional to myocardial viability, was measured at 490 nm, and the level of lactate dehydrogenase (LDH) in the coronary effluent was measured to evaluate the cardiac injury.</p><p><b>RESULTS</b>The pretreatment with ACh (0.1 mol/L, 5 mm) before I/R markedly increased myocardial formazan content, reduced LDH release, improved the recovery of the left veritficular developed pressure, +/- dP/dtmax, and rate pressure product (left ventricular developed pressure multiplied by hean rate) and attenuated the decrease of coronary flow during reperfusion. The opener of MPTP, atiractyloside (20 mmoL/L) or the inhibitor of mitoK(ATP), 5-hydroxydecanoate (100 micromol/L) abolisbed the beneficial effect of ACh.</p><p><b>CONCLUSION</b>In the isolated rat bean, ACh protects myocardium against ischemia/reperfusion injury via inhibiting the opening of MPTP and increasing the opening of mitoKATP in heart.</p>

Study of the effects of Auricularia auricular polysaccharide on local ischemia/reperfusion injury in rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the influence of Auricularia Auricular polysaccharide (APP) on acute cerebral injury induced by ischemia/reperfusion in rats and its underlying mechanism.</p><p><b>METHODS</b>Adult male SD rats were intragastrically pretreated with AAP at a low (50 mg/kg) or high (100 mg/kg) dose once a day for 20 days before operation. Rats intraperitoneally injected with ginkgo biloba extract (EGb671) were taken as positive control. Focal ischemia was achieved by middle cerebral artery occlusion (MCAO) on the right side for 60 min. After 24 hrs of reperfusion, the nerve function defects were recorded by Longa's score and the brain infarct sizes were measured by 2,3,5-Triphenyl-tetrazolium-chlor (TTC) staining. Apoptotic neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining after 48 h of reperfusion. The levels of oxidative stress was determined via the mitochondria-generated reactive oxygen species (ROS).</p><p><b>RESULTS</b>AAP treatment decreased Longa's score, brain infarct size, apoptotic neurons and mitochondria-generated ROS in a dose-dependent manner. AAP at 100 mg/kg gave a better performance compared with EGb671 on all parameters examined.</p><p><b>CONCLUSION</b>AAP treatment protected rat brain from focal ischemia/reperfusion injury by its anti-oxidative effect and worked better than EGb671.</p>

Ginkgo biloba extract protects brain from ischemia/reperfusion injuries / 浙江大学学报·医学版

Ginkgo biloba extract (GBE) is one of the hot spots of drugs extracted from plants recently; it protects brain from ischemia/reperfusion injuries. The mechanism of protective effects includes antioxidation, free radicals clearance, inhibiting the release of excitatory amino acid, anti-inflammation, inhibiting neural apoptosis and other biological effects.

Betulinic acid ameliorates impairment of endothelium-dependent relaxation induced by oxidative stress in rat aorta / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of betulinic acid (BA) on relaxation in isolated rat aortic rings and its antioxidant property on oxidative stress of blood vessels.</p><p><b>METHODS</b>Aortic rings were isolated and BA was cumulatively added into organ bath. Isometric tension of endothelium intact or endothelium denuded thoracic aortic rings previously contracted by phenylephrine (PE) was recorded. Then aortic rings were randomly divided into normal control group, BA control group, H(2)O(2) group and BA+H(2)O(2) group, after being previously contracted by PE, isometric tension of endothelium-dependent relaxation induced by Ach was recorded.</p><p><b>RESULT</b>Exposure of intact endothelium rings previously contracted by PE to BA at the concentrations of 10(-7) mol/L-10(-4) mol/L evoked a significant concentration dependent relaxation, which was inhibited by pretreatment with N omega-nitro-L-arginine methyl ester (L-NAME, 10(-4)mol/L), but not by indometacin (10(-5)mol/L). The pD2 value of BA was 5.24 ± 0.04, and the EC(50)value was 2.45 x 10(-6)mol/L. Exposure of endothelium denuded rings previously contracted by PE to BA did not affect the relaxation in isolated aortic rings. ACh induced a dose-dependent relaxation that was weakened by pretreatment with H(2)O(2) (5 10(-4) mol/L) for 15 min. The EC(50) of BA markedly attenuated the inhibition of relaxation induced by H(2)O(2).</p><p><b>CONCLUSION</b>BA can evoke a concentration-dependent relaxation in aortic rings previously contracted by PE, which may be mediated by NO. And the decrease of endothelium-dependent relaxation in rat aortic rings exposed to H(2)O(2) can be markedly attenuated by BA, which may be mediated by reducing oxidative stress and maintaining the activity of NO in aortic rings.</p>

Partial endothelium-dependent vasorelaxation of crocetin and its mechanism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the vasorelaxation effect of crocetin (CCT) and its mechanism.</p><p><b>METHODS</b>Isolated aortic rings from Sprague-Dawley rats were mounted in the organ bath system. The tension of the aorta was recorded.</p><p><b>RESULT</b>CCT significantly provoked concentration-dependent relaxation in both endothelium-intact and-denuded aortic rings pre-constricted by phenylephrine (10⁻⁵ mol/L), and the vasorelaxation in endothelium-intact aortic rings was stronger than that in endothelium-denuded ones. CCT had no significant effects on aortic rings pre-constricted with KCl (6 × 10⁻² mol/L). Pretreatment with eith L-NAME (10⁻⁴ mol/L), an inhibitor of nitric oxide synthase (NOS), or indomethacin (10⁻⁵ mol/L), an inhibitor of cyclooxygenase, for 30 min significantly attenuated the relaxation of endothelium-intact aortic rings induced by CCT. Besides, both tetraethylammonium (a Ca²(+)-activated K(+) channel inhibitor, 5 × 10⁻³ mol/L) and 4-aminopyridine (a voltage-sensitive K(+) channel inhibitor, 10⁻³ mol/L), but not the ATP-sensitive K(+) channel inhibitor glibenclamide (3 × 10⁻⁶ mol/L), significantly attenuated CCT-induced relaxation in endothelium-denuded aortic rings.</p><p><b>CONCLUSION</b>CCT had partial endothelium-dependent relaxation in rat aortas, which may be mediated by activating the endothelial NOS-NO and cyclooxygenase-prostacyclin pathways. The endothelium-independent relaxation in rat aortas induced by CCT may be mediated by Ca²(+)-activated K(+) channels and voltage-sensitive K(+) channels.</p>