Objective To evaluate the role of silent information regulator fac tor 2-related enzyme 1 (SIRT1)/Forkhead Box O3 (FoxO3a) signaling pathway in berberine pretreatment-induced reduction of hypoxia/reoxygenation (H/R)-caused injury to hepatic parenchymnal cells.Methods Hepatic parenchymal cells obtained from AML12 mice were cultured and seeded in 6-well plates (2 ml/well) and in 96-well plates (200 μl/well) at the density of l×l06 cells/ml.The cells were divided into 4 groups (n=36 each)using a randomn number table:control group (group C),group H/R,berberine pretreatment group (group BP) and SIRT1-siRNA group (group SS).The cells were cultured in normal culture atmosphere (5％ CO2-21％ O2-74％ N2) in group C.In H/R,BP and SS groups,the cells were exposed to hypoxic air (5％ CO2-1％ O2-94％ N2) for 12 h,followed by 6 h reoxygenation in normal culture atmosphere (5％ CO2-21％ O2-74％ N2).In group SS,small interference RNA targeting SIRT1 (SIRT1-siRNA) was added to the culture medium at 24 h prior to hypoxia.Berberine (final concentration 5 μmol/L) was added at 2 h prior to hypoxia in BP and SS groups.At the end of reoxygenation,the cell viability was measured by methyl thiazolyl tetrazolium assay,the malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined using enzyme-linked immunosorbent assay,cell apoptosis was detected by flow cytometry,the expression of SIRT1 and FoxO3α was detected by Western blot,and the acetylation of FoxO3α was measnred by using immunoprecipitation.Apoptotic rate was calculated.Results Compared with group C,the cell viability was significantly decreased,the MDA content was increased,the SOD activity was decreased,apoptotic rate was increased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were increased,and the acetylation of FoxO3α in the nucleus was increased in H/ R,BP and SS groups (P＜ 0.05).Compared with group H/R,the cell viability was significantly increased,the MDA content was decreased,the SOD activity was increased,apoptotic rate was decreased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were increased,and the acetylation of FoxO3α in the nucleus was increased in group BP (P＜0.05).Compared with group BP,the cell viability was significantly decreased,the MDA content was increased,the SOD activity was decreased,apoptotic rate was increased,the expression of SIRT1 and ratio of FoxO3α expression in nucleus/in cytoplasma were decreased,and the acetylation of FoxO3α in the nucleus was decreased in group SS (P＜O.05).Conclusion The mechanism by which berberine pretreatment attenuates H/R-caused injury to hepatic parenchymal cells is related to promotion of SIRT1 expression in cells and inhibition of FoxO3α acetylation in the nucleus.

0.05).The adverse effect of treatment group was significantly less than control group(P

Objective To investigate the clinical characteristics and diagnostic main points of poliomyelitis-like syndrome(PS).Met-hods The clinical data of 15 children with PS were analyzed retrospectively including nervous system manifestation,results of lab andauxiliary examination,diagnosis and reason of misdiagnosis,treatment and progress.Results PS in all 15 children was characterized by mean age of onset(3.3 years),and forerunner infection(in 11 cases,73%)with acute upper respiratory infection of diarrhea 1-2 weeks pre-onset or during onset.All cases were acute flaccid paralysis,12 cases(80%) of which were only one limb.The positive outcome of serologic examination in PS consisted of 4 cases in coxsackie virus-IgM,1 case in EB virus-IgM,1 case in herpes simplex virus-IgM and 1 case in mycoplasma-IgM.All children showed the electromyologram changes in nerve damage.The muscle force of 4 cases increased one grade.Conclusions Children with PS are characterized by the age of onset under 5 years old,acute flaccid paralysis(mostly affected one limb),and the most pathogen being enterovirus.The electromyologram examination can help establish a definite diagnosis in PS.

Objective To explore the risk factors affecting prognosis of children with Tourette syndrome(TS).Methods The follow-up visits were conducted on the clinical data of 98 cases with TS(85 male,13 female;aged 4-16 years old)from 1997 to 2005 in Wuhan children's hospital.All cases were consistent with the diagnostic criteria of TS in the 4th edition of Diagnostic and Statistical Manual of Mental Disorders(DSM-Ⅳ).The investigations were performed by the investigators who received special training using the unified questionnaire with the methods of direct inquiry or by telephone.The factors included sex,age,severity of TS,the primary symptoms,family history,coexisting diseases,basic diseases,perinatal abnormity and family-social relations.The prognosis of TS and these factors were analyzed by linear regression and stepwise regression with SPSS 12.0 software.Results About 16 cases lost follow-up and the other 82 cases with follow-up(72 male and 10 female)received retrospective review.They were 14 to 25 years old with complete data,and 50 cases healed,32 cases not healed.Results from non-conditional simple variant Logistic regression showed that such cases were associated with the following factors:age,family history of TS,severity of TS,coexisting diseases,basic diseases and perinatal abnormity(Pa0.05).Out of 6 suspicious factors,there were coexisting diseases(OR=84.088,95%CI 10.850-651.682),severity of TS(OR=13.956,95% CI 2.412-80.762),and family history of TS(OR=27.127,95% CI 1.047-702.831)of risk factors.Conclusion The long-term prognosis of children with TS may be related with coexisting disease,severity of TS and family history respectively.

In order to establish a method for the determination of the sterols of the oil in the freeze-dried acai (Euterpe oleracea Mart.) and to evaluate its antioxidant activities, a saponification/extraction procedure and high performance liquid chromatography (HPLC) analysis method were developed and validated for the analysis of phytosterols in PEE (Petroleum ether extract). Separation was achieved on a Purosper STAR LP C18 column with a binary, gradient solvent system of acetonitrile and isopropanol. Evaporative light scattering detection (ELSD) was used to quantify β-sitosterol and the total sterols. Peak identification was verified by retention times and spikes with external standards. Standard curves were constructed (r = 0.999 2) to allow for sample quantification. Recovery of the saponification and extraction was demonstrated via analysis of spiked samples. The highest content of total sterols is β-sitosterol. The antioxidant activities of the extracts were evaluated using the total oxyradical scavenging capacity assay (TOSC assay). The result showed that the PEE exhibited significant antioxidant properties, sample concentration and the antioxidant capacity had a certain relevance.
Antioxidants ; analysis ; Arecaceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Phytosterols ; analysis ; Sitosterols ; analysis

Objective To evaluate the efficacy and safety of levetiracetam (LEV) monotherapy on children with epilepsy.Methods Forty-one children (26 cases were male,15 cases were female) with epilepsy aged 7 months to 13 years were treated with LEV as monotherapy.These patients were selected from Department of Neurology ,Wuhan Children′s Hospital, from Aug.2007 to Aug.2009.The starting do-sage of LEV was (13.6?4.7) mg?kg-1?d-1,twice daily,and its objective dosage was (25.7?7.5) mg?kg-1?d-1,twice daily.LEV monotherapy was investigated by a self-controlled and open-label research,and the follow-up period ranged from 6 months to 2 years.Results The effective rate was 68.3% (28 cases),with 39.0% (16 cases) achieving seizure freedom in LEV monotherapy of children with epilepsy.Thirteen patients (31.7%) had poor efficacy in reduction of seizures,7 patients (17.1%) discontinued LEV monotherapy due to either an inadequate seizure control or aggravated seizures.Fifteen patients (36.6%) had the therapy-related adverse events in LEV monotherapy,including gastrointestinal dysfunction (5 cases),irritability (5 cases),dizziness (2 cases) and somnolence (2 cases).The adverse effects appeared in 2-4 weeks of early LEV therapy and were spontaneously disappeared in 1 week to 1 month of continuing therapy.Conclusions The LEV monotherapy is effective and safe for the control of partial and generalized seizures in children with epilepsy.LEV appears to be a broad-spectrum,first-line anti-epileptic drug in treatment of children with epilepsy.

Robust and efficient control of therapeutic gene expression is needed for timing and dosing of gene therapy drugs in clinical applications. Ribozyme riboswitch provides a promising building block for ligand-controlled gene-regulatory system, based on its property that exhibits tunable gene regulation, design modularity, and target specificity. Ribozyme riboswitch can be used in various gene delivery vectors. In recent years, there have been breakthroughs in extending ribozyme riboswitch's application from gene-expression control to cellular function and fate control. High throughput screening platforms were established, that allow not only rapid optimization of ribozyme riboswitch in a microbial host, but also straightforward transfer of selected devices exhibiting desired activities to mammalian cell lines in a predictable manner. Mathematical models were employed successfully to explore the performance of ribozyme riboswitch quantitively and its rational design predictably. However, to progress toward gene therapy relevant applications, both precision rational design of regulatory circuits and the biocompatibility of regulatory ligand are still of crucial importance.
Animals ; Cell Line ; Gene Expression ; Gene Expression Regulation ; Genetic Therapy ; Humans ; Ligands ; Models, Theoretical ; RNA, Catalytic ; genetics ; Riboswitch ; genetics

AIM:To study the expression level of S100 calcium-binding protein A4 (S100A4) in synovial tissue of the knee joint in rheumatoid arthritis (RA) patients and normal persons, and the effect of S100A4 on the angiogenesis induced by rheumatoid arthritis fibroblast-like synoviocytes (RAFLSs).METHODS:The synovial tissue was taken from the knee joint of the RA patients (RA group) and the normal persons (control group).The protein expression of S100A4 and vascular endothelial growth factor (VEGF) in the synovial tissue of the 2 groups was observed by immunohistochemistry.RAFLSs were isolated from synovial tissue of patients with active RA.ELISA was used to detect the effect of S100A4 on the secretion of VEGF by RAFLSs.The effect of S100A4 on the angiogenesis of HUVECs cultured with conditioned medium from RAFLSs was also detected.RESULTS:The protein of S100A4 and VEGF was highly expressed in the synovial tissues of RA group (P<0.05).rhS100A4 significantly stimulated the secretion of VEGF in RAFLSs in a time-and dose-dependent manner (P<0.05).Cultured with conditioned medium from RAFLSs, rhS100A4 significantly promoted HUVECs to form tube-like structures in vitro.CONCLUSION:S100A4 protein is highly expressed in synovial tissue of the knee joint in RA patients, and S100A4 stimulates synovial angiogenesis by promoting RAFLSs to generate VEGF, indicating that S100A4 may be used as a potential target for the treatment of RA.

Abstract: Objective　To analyze the epidemiological characteristics of 151 cases of melioidosis and the drug resistance of Burkholderia pseudomallei (BP), in order to provide the basis for diagnosis, treatment and reasonable prevention of melioidosis. Methods　A total of 151 inpatients and outpatients from the Second Affiliated Hospital of Hainan Medical University from January 1, 2013 to August 31, 2022 were collected, and clinical specimens were submitted for examination to isolate and identify BP strains. The clinical data of 151cases of melioidosis and the drug resistance characteristics of pathogenic bacteria were retrospectively analyzed, and using SPSS26.0 software for statistical analysis. Results　Among 151 cases with BP infection, there were 138 males (91.4%) and 13 females (8.6%); the most patients were aged from 45-<60 years old, accounting for 74 cases (49.0%); melioidosis incidence was concentrated in October (19.2%), November (19.2%), August (9.9%) and July (8.6%), and; the number of confirmed cases showed an increasing trend and the time for confirmation was <10 d; Internal medicine system (31.1%), surgery system (26.5%) and intensive care department (20.5%) were the common departments for treating melioidosis; blood (49.0%), sputum (9.9%) and wound secretion (8.6%) were the main clinical specimens for detecting BP; pulmonary infection (68.2%), sepsis (35.1%) and local suppurative infection (23.8%) were the top clinical manifestations in patients with BP infection; the effective rate of treating melioidosis was 74.8%; abnormal liver function was a risk factor for the curative effect of melioidosis (χ2=5.010, P<0.05); the sensitivity rates of BP strains to sulfamethoxazole-trimethoprim (SXT), doxycycline (DOX), imipenem(IPM), ceftazidime (CAZ), amoxicillin/clavulanate (AMC) and tetracycline (TCY) were generally more than 90%, with sensitivities of 98.7%, 97.2%, 96.7%, 94.0%, 93.2% and 90.7%, respectively. Conclusions　It can be concluded that misdiagnosis or missed diagnosis of melioidosis is easy to occur, and the understanding of the epidemiological characteristics and risk factors in this area should be strengthened. The sensitivity of BP to commonly used antibiotics has shown a certain downward trend, clinical use should be standardized, and drug resistance monitoring should be strengthened to improve the efficacy of melioidosis treatment.

Objective To study the dynamic changes of cytokines including tumor necrosis factor-alpha (TNF-?),interleukin-6 (IL-6),IL-8 in serum and cerebrospinal fluid (CSF) of asphyxia neonates,and to analyze the relationship between cytokines levels and severity of brain damage and neurological outcome. Methods The concentrations of TNF-?,IL-6,IL-8 in serum and CSF were measured by radioimmunoassay in 63 asphyxia neonates. Neurological development was evaluated at 12 months by children′s developmental scale of china.Results The serum concentrations of TNF-?, IL-6,IL-8 were significantly higher in asphyxiated neonates than those in the controls,and they were correlated with the degree of encephalopathy. The level of serum TNF-? was hig-(hest) at the first day and IL-6 was highest at the third day. There was no marked dynamic changes within 5 days in serum IL-8 level. The concentrations of TNF-?,IL-8 in CSF were higher at the first and the third day.The dynamic changes of IL-6 in CSF were similar in serum and they were positively correlated. The serum concentration of IL-6 in severe brain injury group was much higher than those of normal and mild group.The CSF concentration of IL-6 in severe brain injury group was much higher than that of normal group. The CSF concentration of IL-8 in severe brain injury group was much higher than those of the normal and mild group. Conclusions The concentrations of TNF-?,IL-6 and IL-8 are increased both in serum and CSF in asphyxiated neonates which are correlated with severity of hypoxic-ischemic encephalopathy. Cytokine-mediated inflammatory reactions may participate in the mechanism of hypoxic-ischemic brain injury after asphyxiaion.The concentration of IL-6 in serum and IL-6, IL-8 in CSF are correlated with the neurological outcome.