Association of genetic variant and platelet function in patients undergoing neuroendovascular stenting.

INTRODUCTION: The risk of recurrent ischaemic events is related to platelet function, which is often assessed by thromboelastography (TEG). TEG has high interindividual variability. OBJECTIVE: To identify causal variants associated with TEG parameters in patients who receive aspirin and clopidogrel after intra- or extracranial stenting. METHODS: Patients who underwent stenting for extracranial or intracranial stenosis (70-99%) were recruited into the study. Blood samples were obtained for TEG to assess the platelet function before stenting. Aspirin- and clopidogrel-related genetic polymorphisms were determined by the MassARRAY method. Minor allele frequency and Hardy-Weinberg equilibrium (HWE) tests and linkage disequilibrium (LD) analysis were carried out. The influences of genetic polymorphism on TEG parameters were analysed by linear regression. RESULTS: A total of 249 patients were included in this study. Twenty-two selected single nucleotide polymorphisms (SNPs) were genotyped, and no significant deviation from HWE was found for any SNP in the study patients. Four SNPs-rs2104543, rs12772169, rs1998591 and rs1042194-within CYP2C18 were in high LD, and the genetic polymorphisms had a significant impact on the TEG parameters maximal clot strength (MAThrombin) and ADP-induced platelet-fibrin clot strength (MAADP). Patients who carried the loss-of-function CYP2C19*2 (rs4244285) allele were also at risk of increased MAThrombin and MAADP. CONCLUSIONS: Testing for these polymorphisms may be valuable in the identification of patients at high risk of recurrent ischaemic events. Alternative treatments may be considered for these high-risk patients. TRIAL REGISTRATION NUMBER: NCT01925872.

Inhibition of EGFR-induced glucose metabolism sensitizes chondrosarcoma cells to cisplatin.

Chondrosarcomas are malignant cartilage-forming tumors which are resistant to conventional chemotherapy and radiotherapy. By searching in Oncomine which is a cancer microarray database and web-based data mining platform, we found Glut1 and LDHA were upregulated in human chondrosarcoma patient samples. In this study, we reported total epidermal growth factor receptor (EGFR) expression and phosphorylated EGFR were highly activated in human chondrosarcoma cell lines. In addition, overexpression of EGFR contributed to cisplatin resistance. EGFR promoted glucose metabolism of chondrosarcoma cells through the upregulation of glycolysis key enzymes. Interestingly, cisplatin-resistant chondrosarcoma cells showed upregulated glucose metabolism and EGFR signaling pathway. Finally, we demonstrated that the combination of either EGFR inhibitor or anaerobic glycolysis inhibitor with cisplatin showed synergistically inhibitory effects on cisplatin-resistant chondrosarcoma cells through the inducements of apoptosis and cell cycle arrest. Our project proposed a novel function of EGFR in the regulation of glucose metabolism in chondrosarcoma cells and contributed to the development of therapeutic strategies for the clinical treatment of chondrosarcoma patient.

Protective effect of melatonin on bone marrow mesenchymal stem cells against hydrogen peroxide-induced apoptosis in vitro.

Bone marrow mesenchymal stem cells (MSCs) transplantation has shown great promises for treating various central nervous system (CNS) diseases. However, poor viability of transplanted MSCs in injured CNS has limited the therapeutic efficiency. Oxidative stress is one of major mechanisms underlying the pathogenesis of CNS diseases and has a negative impact on the survival of transplanted MSCs. Melatonin has recently been reported to have the antioxidant and anti-apoptotic properties in serial of cells. This study was designed to investigate the protective effect and potential mechanisms of melatonin against hydrogen peroxide (H2O2)-induced apoptosis of MSCs. MSCs were pretreated with melatonin (1, 10, and 100 nM, respectively) for 30 min, followed by exposure to 400 µM H2O2 and melatonin together for 12 h. The present study reports that melatonin pretreatment significantly attenuated H2O2-induced MSC apoptosis in a dose-dependent manner. Consistently, melatonin effectively suppressed the generation of intracellular ROS, expression ratio of Bax/Bcl-2, activation of caspase-3 and expression of phospho-P38MAPK in H2O2-induced MSCs. Luzindole, a nonselective melatonin receptor antagonist, significantly counteracted melatonin's promotion effect on cell survival, indicating that melatonin exerts its protective effect on MSCs, at least in part, through the activation of melatonin receptors. The findings suggest that melatonin may be an effectively protective agent against oxidative stress-induced MSC apoptosis.

Characteristics of traffic flow at nonsignalized T-shaped intersection with U-turn movements.

Most nonsignalized T-shaped intersections permit U-turn movements, which make the traffic conditions of intersection complex. In this paper, a new cellular automaton (CA) model is proposed to characterize the traffic flow at the intersection of this type. In present CA model, new rules are designed to avoid the conflicts among different directional vehicles and eliminate the gridlock. Two kinds of performance measures (i.e., flux and average control delay) for intersection are compared. The impacts of U-turn movements are analyzed under different initial conditions. Simulation results demonstrate that (i) the average control delay is more practical than flux in measuring the performance of intersection, (ii) U-turn movements increase the range and degree of high congestion, and (iii) U-turn movements on the different direction of main road have asymmetrical influences on the traffic conditions of intersection.

[Prognostic correlation of intracranial pressure monitoring in patients with severe craniocerebral injury].

OBJECTIVE: To explore the clinical application of intracranial pressure (ICP) monitoring and its prognostic correlation in patients with severe craniocerebral injury. METHODS: A total of 216 severe craniocerebral injury patients with scores of Glasgow coma scale 3-8 underwent craniotomy at Affiliated Qilu Hospital, Shandong University.And 168 cases of ICP monitoring were divided into 3 treatment groups and another 48 cases without ICP monitoring selected as the control group.According to ICP, stepwise treatment was administered to control the level of ICP and maintain the cerebral perfusion pressure to analyze the relationship between ICP monitoring and prognosis. RESULTS: As compared with the control group, there were significant decreases of disability and mortality rate for patients with ICP monitoring (A, B, C group). Especially group C had a better prognosis than the other groups for statistical significance.In addition, the dose and duration of mannitol of group A, B or C were significantly lower than those of the control group (P < 0.05). CONCLUSION: The application of ICP monitoring is capable of reducing mortality, improving prognosis and enhancing success rate of treating severe craniocerebral injury.

Dynamic Changes of ORF1ab and N Gene Ct Values in COVID-19 Omicron Inpatients of Different Age Groups - Beijing Municipality, China, November-December 2022.

Introduction: In November 2021, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant was identified as the variant of concern and has since spread globally, replacing other cocirculating variants. To better understand the dynamic changes in viral load over time and the natural history of the virus infection, we analyzed the expression of the open reading frames 1ab (ORF1ab) and nucleocapsid (N) genes in patients infected with Omicron. Methods: We included patients initially admitted to the hospital for SARS-CoV-2 infection between November 5 and December 25, 2022. We collected daily oropharyngeal swabs for quantitative reverse transcriptase-polymerase chain reaction tests using commercial kits. We depicted the cycle threshold (Ct) values for amplification of ORF1ab and N genes from individual patients in age-specific groups in a time series. Results: A total of 480 inpatients were included in the study, with a median age of 59 years (interquartile range, 42 to 78; range, 16 to 106). In the <45-year-old age group, the Ct values for ORF1ab and N gene amplification remained below 35 for 9.0 and 11.5 days, respectively. In the ≥80-year-old age group, the Ct values for ORF1ab and N genes stayed below 35 for 11.5 and 15.0 days, respectively, which was the longest among all age groups. The Ct values for N gene amplification took longer to rise above 35 than those for ORF1ab gene amplification. Conclusion: The time to test negative varied among different age groups, with viral nucleic acid shedding taking longer in older age groups compared to younger age groups. As a result, the time to resolution of Omicron infection increased with increasing age.

Pharmacokinetic/pharmacodynamic studies on exenatide in diabetic rats.

AIM: To quantitatively evaluate the blood glucose-lowering effect of exenatide in diabetic rats. METHODS: Male Harlan-Sprague-Dawley rats were treated with high-fat diet/streptozotocin to induce type 2 diabetes. After subcutaneous administration of a single dose of exenatide (4.2, 42, or 210 µg/kg), serum exenatide, insulin concentration and blood glucose were measured. The pharmacokinetics of exenatide was characterized by a two-compartment model with first-order absorption. Insulin turnover was characterized by an effect compartment and indirect response combined model. Glucose turnover was described using an indirect response model with insulin (in effect compartment) stimulating glucose disposition and insulin (in insulin compartment) inhibiting glucose production simultaneously. The model parameters were estimated using nonlinear mixed-effects model program. Visual predictive check and model evaluation were used to make assessments. RESULTS: Exenatide exhibited rapid absorption with k(a)=4.45 h(-1), and the two-compartment model well described its pharmacokinetic profile. For the pharmacodynamic model, exenatide increased insulin release with the estimated S(m1) of 0.822 and SC(50) of 4.02 µg/L. It was demonstrated that insulin stimulated glucose dissipation (S(m2)=0.0513) and inhibited the production of glucose (I(m)=0.0381). Visual predictive check and model evaluation study indicated that a credible model was developed. CONCLUSION: The glucose-lowering effect of exenatide in diabetic rats is reliably described and predicted by the combined effect compartment/indirect response model.

[Aberrant methylation of NF2, TIMP-3 and THBS1 genes and their diagnostic values in meningiomas].

OBJECTIVE: To explore the functions of NF2, TIMP-3 and THBS1 genes in the tumorigenesis or progression of meningiomas and analyze the values of these genes in early diagnosis, therapy and prognostic evaluation in meningiomas. METHODS: A total of 66 cases with histological sections of meningiomas, including solitary (SMs, n = 30) and multiple meningiomas (MMs, n = 36), were retrieved from our departmental archives. All cases were regrouped as benign, atypical and anaplastic (malignant) by hematoxylin & eosin staining according to the recently published WHO classification of nervous system tumors. Genomic DNA was extracted from tumor sections and methylation-specific polymerase chain reaction (MSP) performed to detect the CpG methylation status. Normal brain tissue was used as the control group. And then the differences of methylation rate between SMs and MMs tissues and among different subgroups were analyzed by statistical analyses. RESULTS: The results of methylation in different types of meningiomas demonstrated that the rates of NF2, TIMP-3 and THBS1 methylation were 26.7% (8/30), 16.7% (5/30) and 36.7% (11/30) in 30 SMs tissues and 30.6% (11/36), 22.2% (8/36) and 22.2% (8/36) in 36 MMs tissues respectively. But no aberrant methylation of NF2, TIMP-3 and THBS1 genes was found in normal brain tissue. No significant differences in three types of gene methylation rates existed between SMs and MMs in the I-III grade meningiomas. Nevertheless, there was great difference between grades I, II and III in SMs and MMs while no significant difference was found between grades II and III. CONCLUSION: The methylation of NF2, TIMP-3 and THBS1 is correlated with the tumorigenesis of meningiomas (grade II and III). As an important pathogenetic cause of meningiomas, it may be used as a clinical tool for an early diagnosis of meningiomas.

[Expression and significance of MMP-9, PR, Ki-67 and Survivin in multiple meningiomas].

OBJECTIVE: To detect the expressions of matrix metalloproteinase-9(MMP-9), progesterone receptor(PR), Ki-67 and Survivin in the multiple meningiomas (MMs) and explore its genesis, diagnosis and bionomics features. METHODS: A total of 66 cases with histological sections of meningiomas retrieved from the archives of Department of Pathology of our hospital, including 30 cases of solitary meningiomas (SMs) and 36 cases MMs, were regrouped as benign, atypical and malignant by hematoxylin and eosin staining according to the World Health Organization classification of nervous system tumors. Immunohistochemistry was performed to detect the expressions of MMP-9, PR, Ki-67 and Survivin in 36 cases of MMs and 30 cases of SMs. And normal brain tissue was selected as a control group. The staining intensity was analyzed quantitatively for the differential expressions of MMP-9, PR, Ki-67 and Survivin between SMs and MMs. RESULTS: No expression of MMP-9, PR, Ki-67 and Survivin was detected in 5 normal brain tissues, but the expression rates were 100%, 53%, 23% and 88% respectively for significant difference comparing with normal tissue. The result of statistical analysis showed that there was significant difference in the expression intensity of MMP-9 and PR between two groups. The expression intensity MMP-9 in multiple group was significantly higher in MMs than that in SMs (P < 0.01) while PR was lower in MMs than that in SMs (P < 0.05). But no significant difference was found for the expression of Ki-67 or Survivin between two groups. CONCLUSION: The detections of MMP-9, PR, Ki-67 and Survivin are helpful in the clinical diagnosis and early detection of meningioma.

[Clinical experiences of normal saline pressed injection via lumbar puncture in the treatment of apnea due to acute tonsillar hernia].

OBJECTIVE: To summarize the clinical experiences of normal saline pressed injection via lumbar puncture in the treatment of acute tonsillar hernia induced apnea. This procedure was routinely carried out after external ventricular drainage and/or lesion removal via open craniotomy. METHODS: During the period of 1969 to 2005, a total of 43 patients failed to regain respiratory after external ventricular drainage using rapid small hole cranio-puncture apparatus or lesion removal via open craniotomy. They underwent lumbar puncture and normal saline was pressed injected via a lumbar puncture needle. The patient data were retrospectively analyzed. RESULTS: Eleven of 43 patients had spontaneous respiration and fully recovered (25.6%), 16 patients regained respiration but died eventually (37.2%) and 16 patients failed to regain respiration (37.2%). The effective rate was 62.8%. CONCLUSION: For the patients failing to regain respiration after external ventricular drainage or supratentorial lesion removal via open craniotomy, the conservative treatment should not be the first choice. The pressed injection of normal saline via lumbar puncture may rescue some patients.

Impact of Azithromycin on Forsythiaside Pharmacokinetics in Rats: A Population Modeling Method.

OBJECTIVE: Lianhuaqingwen and Shuanghuanglian are drug treatment options for Corona Virus Disease 2019 (COVID-19). In China, use of traditional Chinese medicine with Shuanghuanglian or Lianhuaqingwen (for them, forsythiaside is the active antiviral and antibacterial component) in combination with azithromycin is common for the treatment of pediatric pneumonia. It is important to understand the reason why the combination of these compounds is better than a single drug treatment. This study aimed to explore the pharmacokinetic interaction between forsythiaside and azithromycin. METHODS: Twelve male Sprague-Dawley rats were randomly divided into an experimental group (Forsythia suspensa extract and azithromycin) and a control group (a single dose of Forsythia suspensa extract in 5% glucose solution). Plasma samples were collected at scheduled time points, and the high-performance liquid chromatography combined with ultraviolet method was used to determine the plasma forsythiaside concentration. Non-compartmental analysis and population pharmacokinetic methods were used to investigate the forsythiaside pharmacokinetic difference between the experimental and control group. RESULTS: Compared with a single administration, the area under the curve and half-life of forsythiaside increased, and forsythiaside clearance decreased significantly after co-administration with azithromycin. The in vivo behavior of forsythiaside could be described by the one compartment model. The forsythiaside clearance decreased when combined with azithromycin. Visual evaluation and bootstrap results suggested that the final model was precise and stable. CONCLUSION: Co-administration of azithromycin can significantly decrease the forsythiaside clearance and increase drug exposure. A lower dose of azithromycin can obtain sufficient forsythiaside concentration to provide antiviral and antibacterial activity.

Rescue of social deficits by early-life melatonin supplementation through modulation of gut microbiota in a murine model of autism.

Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder with a rapidly increasing global prevalence. Early unstable and immature microbiota are often observed in ASD patients, resulting in neurobehavioral dysfunction. Since the establishment of stable gut microbiota in early life falls into the same critical time window as neurodevelopment, manipulations of the gut microbiota during early life could become a promising strategy for ASD. Melatonin is an endogenous hormone and can restore gut microbial dysbiosis under various disease conditions. Here, we explored the effects of melatonin supplementation during early life on the gut microbiota of the offspring and the subsequent impact on ASD-associated behaviors. Using the valproic acid (VPA) - induced mouse model of autism, we found that melatonin supplementation during late gestation and early postnatal development rescued the social deficits of the offspring. In addition, melatonin restored gut microbial dysbiosis in the VPA-exposed offspring, which was characterized by the significant upregulation of Akkermansia spp. Furthermore, supplementation of Akkermansia spp. alleviated the social deficits induced by VPA exposure via activating the dopaminergic neurons in the ventral tegmental area. These findings discover a novel mechanism underlying the gut microbiota regulation of social behaviors and provide the biological basis for developing gut microbiota-based therapeutics for ASD.

The clinical population pharmacokinetics, metabolomics and therapeutic analysis of alkaloids from Alstonia scholaris leaves in acute bronchitis patients.

BACKGROUND: Capsule of alkaloids from leaf of Alstonia scholaris (CALAS) is a new investigational botanical drug (No. 2011L01436) for respiratory disease. Clinical population pharmacokinetics (PK), metabolomics and therapeutic data are essential to guide dosing in patients. Previous research has demonstrated the potential therapeutic effect of CALAS on acute bronchitis. Further clinical trial data are needed to verify its clinical efficacy, pharmacokinetics behavior, and influence of dosage and other factors. PURPOSE: To verify the clinical efficacy and explore the potential biomarkers related to CALAS treatment for acute bronchitis. MATERIALS AND METHODS: Oral CALAS was assessed in a randomized, double-blind, placebo-controlled trial. Fifty-five eligible patients were randomly assigned to four cohorts to receive 20, 40 or 80 mg, of CALAS three times daily for seven days, or placebo. Each CALAS cohort included 15 subjects, and the placebo group included 10 subjects. A population PK model of CALAS was developed using plasma with four major alkaloid components. Metabolomics analysis was performed to identify biomarkers correlated with the therapeutic effect of CALAS, and efficacy and safety were assessed based on clinical symptoms and adverse events. RESULTS: The symptoms of acute bronchitis were alleviated by CALAS treatment without serious adverse events or clinically significant changes in vital signs, electrocardiography or upper abdominal Doppler ultrasonography. Moreover, one compartment model with first-order absorption showed that an increase in aspartate transaminase will reduce the clearance (CL) of scholaricine, and picrinine CL was inversely proportional to body mass index, while 19-epischolaricine and vallesamine CL increased with aging. The serum samples from acute bronchitis patients at different time points were analyzed using UPLC-QTOF in combination with the orthogonal projection to latent structures-discriminant analysis, which indicated higher levels of lysophosphatidylcholines, lysophosphatidylethanolamines and amino acids with CALAS treatment than with placebo. CONCLUSION: This is the first study to evaluate the clinical efficacy and explored the potential biomarkers related to CALAS therapeutic mechanism of acute bronchitis by means of clinical trial combined the metabolomics study. This exploratory study provides a basis for further research on clinical efficacy and optimal dosing regimens based on pharmacokinetics behavior. Additional acute bronchitis patients and CALAS PK samples collected in future studies may be used to improve model performance and maximize its clinical value.

Expression of the Golgi phosphoprotein-3 gene in human gliomas: a pilot study.

The oncogene Golgi phosphoprotein 3 (GOLPH3) has been found in several solid cancers, but its expression in glioma tumor tissues is unknown. Reverse transcription polymerase chain reaction and Western blot was used to investigate expression of GOLPH3 mRNA and protein, respectively, in 76 patients with glioma. Non-cancerous brain issues and lung cancer cells were used as controls. There were 45 males and 31 females (mean age 50.7 ± 12.8 years). Astrocytoma was found in 65 patients and glioblastoma in 11. No GOLPH3 expression was found in the non-cancerous brain tissues, but positive GOLPH3 protein was found in lung cancer cells. GOLPH3 mRNA and protein expression were identified in 40 patients with glioma (52.6%). Positive expression of GOLPH3 mRNA or protein was similar in patients with astrocytoma grades I-III and glioblastoma (P > 0.05). The highest mean value of GOLPH3 mRNA and protein was found in patients with glioblastoma (P < 0.01) whereas the lowest mean values were found in those with grade I astrocytoma (P < 0.01). We concluded that, in this pilot study, GOLPH3 expression was present in more than half of the patients with glioma. The amount of GOLPH3 expression in the glioma was associated with the severity of the tumor. Whether positive GOLPH3 gene expression can be used as a predictor for prognosis of the patients or as a therapeutic target for glioma requires further investigation.

Association between AKT/mTOR signalling pathway and malignancy grade of human gliomas.

The mammalian target of rapamycin (mTOR) signaling pathway has emerged as a major effector of cell growth and proliferation, and is an attractive target for cancer therapy. However, the association between mTOR pathway and the malignancy grade of human gliomas has not been thoroughly investigated. Tumor tissues from 87 Chinese patients (49 males, average age of 51.7 ± 13.0 years, range 15-78) with glioma were prospectively collected. The expression of three key proteins of the mTOR pathway, pAKT, pmTOR and p-p70S6 kinase (p-p70S6K) was measured by semi-quantitative immunohistochemical techniques. Grade I-II, III and IV glioma was pathologically identified in 27 (31.0%), 24 (27.6%) and 36 (41.4%) patients, respectively. Of the 87 patients, pAKT, pmTOR and p-p70S6K were found in 63 (72.4%), 65 (74.7%), and 63 (72.4%) patients, respectively. The expression of all three pAKT, pmTOR and p-p70S6K proteins was found in 42 (48.3%) patients, while only one or two of the three proteins were found in the remaining patients (51.7%). The percentage of patients with very strong expression of pAKT, pmTOR and p-p70S6K in grade IV glioma was 13 (36.1%), 16 (44.4%) and 15 (41.7%), respectively, which was greater than in grade I or II tumors (0-3.7%, P < 0.01). In conclusion, expression of mTOR pathway proteins pAKT, pmTOR and p-p70S6K can be found in human glioma of all malignancy grades. However, higher levels of these proteins were associated with advanced malignancy grades of the tumor.

[Neuroendoscope-assisted surgical treatment of spinal dural arteriovenous fistulas].

OBJECTIVE: To study the utility of neuroendoscope-assisted surgery in the treatment of spinal dural arteriovenous fistulas. METHODS: From November 2008 to November 2010, 8 cases of spinal dural arteriovenous fistulas underwent neuroendoscope-assisted surgical treatment by a hemilaminectomy approach. Retrospective analyses were performed for their clinical manifestations, imaging findings, surgical approaches, postoperative recovery and follow-up profiles. RESULTS: All were of single fistula. Under the assistance of neuroendoscope, the fistulas were found intra-operatively and the draining veins disconnected successfully. The results of post-operative angiography showed the disappearance of all draining veins. After a follow-up period of 3 - 35 months, 2 cases became asymptomatic, 5 cases improved obviously and 1 case had no change. CONCLUSION: Neuroendoscope-assisted surgery is mini-invasive, safe and effective in the treatment of spinal dural arteriovenous fistulas.

Down-regulating microRNA-20a regulates CDH1 to protect against cerebral ischemia/reperfusion injury in rats.

Studies have extensively focused on the involvement of microRNAs (miRNAs) in cerebral ischemia/reperfusion (I/R) injury but not much on the specific role of miR-20a. Hence, this study is purposed to decipher whether miR-20a could regulate cadherin 1 (CDH1) to affect cerebral I/R injury in rats. Rat transient middle cerebral artery occlusion model (MCAO) was established. Rats were injected with lentiviral solution containing miR-20a inhibitor, or overexpressed CDH1 or combined depleted miR-20a and CDH1 to explore their roles in cerebral I/R injury. Oxidative stress-related factors, miR-20a, CDH1, nuclear factor-kappaB (NF-κB) and Nestin expression in brain tissues were detected by RT-qPCR and western blot assay. The target relation between miR-20a and CDH1 was predicted by online website and further confirmed by luciferase activity assay. In rats with cerebral I/R injury, increased miR-20a and decreased CDH1 were found in brain tissues. Reduction of miR-20a or elevation of CDH1 attenuated behavior function in MCAO rats. Inhibiting miR-20a or restoring CDH1 restrained oxidative stress, attenuated pathological damage of neurons, promoted neuron survival, and down-regulated NF-κB and Nestin expression in brain tissues of MCAO rats. CDH1 was determined to a target gene of miR-20a. This study elucidates that down-regulating miR-20a elevates CDH1 to protect neurons from cerebral I/R injury, which paves a new way for treatment of cerebral I/R injury.

[The production of hydroxyl radical in HRP-NADH-H2O/O2 systems and its application in chlorobenzene removal].

The mechanism of radical generation in HRP-NADH-O2/H2O2 systems and state-change of horseradish peroxidase (HRP) was investigated by using ESR and UV measurements, and the novel enzyme-coenzymatic systems were performed to degrade chlorobenzene as a non-phenolic persistent organic pollutants. The UV results showed that compound III was produced from HRP oxidized by hydrogen peroxide with the catalysis of NADH, which would generate hydroxyl radical. The ESR results demonstrated the production of *OH and O2-. in enzyme-coenzymatic system in the presence of O2 or H2O2 with DMPO and POBN as spin-trappers, respectively. In HRP-NADH-H2O2 system, compound III was the main state of HRP in the initial 10 min, and then converted to HRP with generating hydroxyl radical; and after the addition of oxygen, the production of hydroxyl radical was promoted rapidly, as 4 times as that of the system in absence of oxygen. The addition of SOD(Zn-Cu) decreased the production of hydroxyl radical significantly, resulting from that SOD eliminated O2 reduction to O2-. by NADH and then inhibited *OH formation. The results showed that NADH could improve by about 20% enzyme activity of HRP for phenol removal. The removal of chlorobenzene with HRP-NADH-H2O2 and HRP-NADH-H2O2-O2 systems reached 24.6% and 48.2%, respectively, which was much higher than that with traditional enzymatic system (1.42%), showing a promising prospect in proposal

[The methylation analysis of EMP3 and PCDH-gamma-A11 gene in human glioma].

OBJECTIVES: To study the relationship between promoter methylation and mRNA expressions of EMP3 and PCDH-gamma-A11 genes in human glioma, and to analyze the regulation mechanism of promoter methylation in the progression of glioma. METHODS: The promoter methylation of EMP3 and PCDH-gamma-A11 was studied by a methylation specific PCR in 88 primary astrocytoma, 10 normal brain tissues and 2 glioma cell lines. The mRNA expressions were detected by real-time PCR in 30 primary glioma and 10 normal brain tissues. The correlations of their promoter methylation, mRNA expressions and clinicopathologic characteristics were analyzed. The promoter methylation were also detected in U251 and SHG-44 cell lines. RESULTS: The promoter methylation of EMP3 was detected in 42 tumors (47.7%) and the methylation of PCDH-gamma-A11 was detected in 76 tumors (86.4%). Their mRNA expressions were all significantly decreased in different pathological grade astrocytomas compared to the normal brain tissues (P < 0.01). Their expressions were suppressed but could be reactivated by 5-aza-deoxycytidine in U251 and SHG-44 cell lines. CONCLUSIONS: The promoter methylation of EMP3 and PCDH-gamma-A11 genes may lead to the down-regulation of their mRNA levels in glioma. The promoter methylation and mRNA expressions of EMP3 and PCDH-gamma-A11 are closely related with the malignant development of glioma. The promoter methylation of the two genes may provide clues to evaluation of glioma malignancy as well as its prognosis. It also gives us an insight for future glioma medical therapy with a demethylating agent.

Pharmacokinetic interaction of Forsythia suspensa extract and azithromycin injection after single and co-intravenous administration in rats.

Azithromycin and Chinese medicine forsythia are often used together to treat pediatric mycoplasma infections in China. We aimed to investigate the pharmacokinetic interaction of Forsythia suspensa extract and azithromycin after single and co-intravenous administration in rats. Male Sprague-Dawley rats received single (Forsythia suspensa extract or azithromycin) treatment or co-administration of Forsythia suspensa extract and azithromycin. Blood samples were collected at scheduled times, and drug concentrations were determined by HPLC-UV or HPLC-MS/MS methods. Both non-compartmental analyses and nonlinear mixed-effects modeling approaches were applied to fit pharmacokinetic data and evaluate the impact of co-administration. Pharmacokinetic analysis showed that the area under the curve of azithromycin and forsythiaside increased, and clearance decreased significantly (P < 0.05), after co-administration. The in vivo behavior of both azithromycin and forsythiaside could be appropriately described by the two-compartmental model. The final population pharmacokinetic model indicated that co-administration decreased the central volume of azithromycin and forsythiaside clearance significantly. Co-administration of Forsythia suspensa extract and azithromycin significantly decreased the clearance and increased exposure for both drugs. Pharmacokinetic data suggest that drug co-administration may increase efficiency.