PDE4 inhibition protects against rotenone-induced neurotoxicity through activation of the AMPK/Sirt1 pathway / 中国药理学与毒理学杂志

OBJECTIVE We have previously shown that inhibition of phosphodiesterase-4(PDE4)protects against neuronal damage in models of Parkinson's dis-ease(PD).However,the mechanisms have not yet been completely revealed.Here we aimed to elucidate the pharmacological effects and mechanisms of action of rof-lupram(ROF),an novel PDE4 inhibitor,in experimen-tal models of PD.METHODS The survival rate,apopto-sis rate and toxicity level of SH-SY5Y cells were deter-mined by MTT,flow cytometry and lactate dehydroge-nase detection kit.At the same time,LYT staining was used to detect the changes of lysosome fluorescence intensity:Western blotting was used to detect the changes of lysosome associated proteins,Sirtuin1 and α-Syn;NAD/NADH assay kit was used to determine the change of NAD content.To explore whether SIRT1 inhibitor(EX527)and lysosomal inhibitor could block the effect of ROF.In addition,ROT was used to stimulate C57BL/6J mice to construct a mouse model of PD to verify the effect and mechanism of ROF.The changes of motor function were evaluated by behavioral experiments(pole climb-ing,bar rotating and balance beam experiments).Super-oxide dismutase kit and Western blotting were used to detect the changes of SOD activity and expression of related proteins in substantia nigra.RESULTS We showed that pretreatment with ROF significantly attenu-ated cell apoptosis in ROT-treated SH-SY5Y cells.Fur-thermore,ROF significantly enhanced the lysosomal function,as evidenced by the increased levels of mature cathepsin D(CTSD)and lysosomal-associated mem-brane protein 1(LAMP1)through increasing NAD+/NADH and the expression of sirtuin 1(SIRT1).Pretreatment with an SIRT1 inhibitor selisistat(SELI,10 μ mol·L-1)attenuated the neuroprotection of ROF,and ROF-increased expression levels of LAMP1 and mature CTSD.Moreover,inhibition of CTSD by pepstatin A(20 μmol·L-1)attenuated the protective effects of ROF.In vivo study was conducted in mice exposed to ROT(10 mg·kg-1·d-1,ig)for six weeks;then,ROT-treated mice received ROF(0.5,1 and 2 mg·kg-1·d-1,ig)for four weeks.ROF significantly ameliorated motor deficits,which was accompanied by increased expression levels of tyro-sine hydroxylase,SIRT1,mature CTSD,and LAMP1 in the substantia nigra pars compacta.CONCLUSION Taken together,these results demonstrate that ROF exerts a neuroprotective action in PD models.The mech-anisms underlying ROF neuroprotective effects appear to be associated with NAD+/SIRT1-dependent activation of lysosomal function.

Cataract surgery and intraocular lens power calculation after radial keratotomy: analysis of 8 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the clinical characteristics of cataract surgery after radial keratotomy (RK) and appropriate calculation of intraocular lens (IOL) power.</p><p><b>METHODS</b>Eight patients with cataract (12 eyes) after RK were treated in our hospital from March, 2010 to June, 2013. The visual acuity, keratometric power and length of the ocular axis were examined before the operation. For each patient, 3 groups of corneal curvature values were measured using a automatic keratometer (TOPCON-KR8800) and the minimal K value was selected. Myopic or hyperopic posterior chamber IOL (-1.00 to -2.00 D) were selected based on automatic calculations with SRK-T. Phacoemulsification and IOL implantation were then performed, and the patients were followed up for visual acuity and refractive statuses at 3 months after the operation.</p><p><b>RESULTS</b>All the 12 operated eyes showed improved visual acuity after the operation. The uncorrected visual acuity reached 0.8 to 1.0 in 6 eyes and 0.4 to 0.6+ in the other 6, with a corrected visual acuity ranging from 0.6 to 1.0. The refractive status after operations was nearly emmetropic (+0.75 to -1.00 D) in 6 eyes and myopic in the other 6 (-1.00 to -2.50 D).</p><p><b>CONCLUSIONS</b>Phacoemulsification and IOL implantation is feasible in cataract patients with previous RK. Selecting the minimal K values for central corneal curvature and calculation of the IOL power using the SRK T equation with a reservation of -1.00 to -2.00 D can better ensure the safety of the procedure and avoid the occurrence of hyperopia >+3.00D.</p>

Analysis of stability-related risk factors for carotid plaques in patients with type 2 diabetes mellitus / 中国医师杂志

Objective To investigate the carotid plaque conditions and clinical commonly used test events in patients with type 2 diabetes mellitus (T2DM) and to explore the risk factors affecting the stability of plaques in carotid artery.Methods According to the results of cervical vascular color Doppler ultrasound examination, 125 patients with type 2 diabetes mellitus were divided into unstable plaque group (n=21), stable plaque group (n =54), and non-plaque group (n =50).Analysis related results including age, gender, systolic blood pressure (SBP), diastolic blood pressure (DBP), intima-media thickness (IMT), fasting blood glucose (FBG), triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), uric acid (UA), fibrinogen (Fbg), D-dimer (D-D), free triiodothyronine (FT3), free thyroxine (FT4), and thyroid stimulating hormone (TSH) were analyzed.Results Age, gender, SBP, DBP, TG, HDL, D-D, and TSH were without significant differences among groups.Glucose and Fbg were significant different (P ＜ 0.05) between unstable plague group and stable plaque group/no plaque group.No significance difference was found between stable and no plaque groups.IMT, TC, LDL, and FT4 were significant different (P ＜ 0.05) between unstable and stable plaque groups, and between stable and no plaque groups.Logistic multiple regression analysis suggested that IMT and TC might be independent risk factors for the stability of plaques in carotid artery of patients with type 2 diabetes mellitus (P ＜ 0.05).Conclusions The factors affecting formation of plaques in carotid artery include glucose level, TC, and LDL in patients with type 2 diabetes mellitus.The factors affecting its stability include thickness of IMT, TC, LDL, and FT4.IMT and TC were the independent risk factors for the stability of plaques in carotid artery.

Evaluation of a novel fully automated real-time PCR assay for hepatitis B virus DNA quantification / 中华检验医学杂志

Objective To evaluate the analytical performance of a novel HBV DNA assay based on automated DNA extraction and real-time fluorescence quantitative PCR .Methods Analytic verification studies.Accuracy and lower limit of detection were assessed by determining a panel of HBV standard plasma of WHO.HBV standard plasma (genotype A, B, C and D) at 6 different concentrations were measured 18 times to evaluate precision and reproducibility .Pseudo-viral particles at high HBV DNA concentration were serially diluted to assess linear range .One hundred and forty-four clinical specimens were quantified for HBV DNA so as to evaluate the correlation between the new test and COBAS ? system. Results Quantification of HBV standard plasma showed acceptable accuracy , with each deviation between observed and expected values within ±0.35 lg IU/ml (-0.17-0.32 lg IU/ml).Intra-assay coefficients of variation ( CV) for genotype A , B, C and D were 3.87% -6.32%, 0.45% -14.68%, 0.16% -8.36% and 0.64%-13.01%respectively, and the inter-assay CV were 5.67%-9.69%, 1.28%-15.68%, 0.36%-9.05%and 1.69%-13.65%, separately.Linearity assessment exhibited an excellent dynamic range of linear quantification from 20 to 1.0 ×1010 IU/ml ( r =0.998, P <0.001 ) .And the satisfactory results obtained at 3 levels of HBV DNA concentration (10, 20, 50 IU/ml, respectively) confirmed the claimed lower limit of detection with 5/5 detectable rate at 20 IU/ml.Furthermore, good correspondence was observed between the new HBV DNA assay and the COBAS ? system with 100% ( 144/144 ) qualitative coincidence and significant correlation based on 104 positive data ( r=0.984, P<0.000 1).Conclusions The novel fully-automated real-time PCR assay displayed good analytical and clinical performance for highly sensitive detection of HBV DNA.It was well suited for monitoring antiviral responses as well as drug resistance according to current clinical practice guidelines for the management of chronic HBV infection .

Clinical significance of serum myocardial enzymes and CSF LDH detection in adults with intracranial infection / 中国医师杂志

Objective To investigate the changes and clinical significance of serum lactate dehydrogenase (LDH),creatine kinase (CK),glutamate pyruvate transaminase (AST),and cerebrospinal fluid lactate dehydrogenase (CSF LDH) in adult patients with acute central nervous system infection.Methods The levels of myocardial enzymes (AST,LDH,and CK) in serum of 96 adult patients with acute intracranial infection in 7days and 39 healthy people were measured by Beckman automatic biochemical analyzer and enzyme rate assay,and CSF LDH level in 96 patients were measured simultaneously.Results (1) The serum myocardial enzymes (LDH,CK,and AST) of intracranial infection group (47 cases with viral encephalitis,30 cases with tuberculous meningitis,and 19 cases with purulent encephalitis) were significantly higher than those of normal control group (P ＜0.01).(2)The myocardial enzymes (LDH,and AST ) of patients with cerebral functional disorder were significantly higher than those of patients with normal cerebral function (P ＜0.05).(3)The levels of serum AST,LDH,and CK in the virus encephalitis group,serum AST and LDH in the purulent encephalitis group,and serum LDH in the tuberculous meningitis group were significantly higher than those in the control group (P ＜ 0.01).The CSF LDH level in the viral meningitis group was prominently lower than that in the tuberculous encephalitis group and purulent encephalitis group,respectively (P ＜0.01).(4) No correlations were found between CSF LDH and serum myocardial enzymes (P ＞0.05).Conclusions (1)There is significant change in the levels of serum LDH,CK,AST,and CSF LDH of adult patients with acute intracranial infection,especially in infected patients with cerebral functional disorder,and the change of LDH is the most obvious.(2)The levels of serum myocardial enzymes and CSF LDH are helpful to the differential diagnosis of intracranial infection in early stage,and judging the severity of the illness.

A preliminary assessment of the clinical utility of measuring hepatitis C virus antibody to evaluate infection status / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the potential of hepatitis C virus (HCV) antibody measurement as a clinical approach to determine the infection status and potential for spontaneous-resolution among patients with HCV mono-infection and HCV/human immunodeficiency virus (HIV) co-infection.</p><p><b>METHODS</b>A total of 340 individuals who tested positive for serum anti-HCV antibodies and/or serum anti-HW antibodies were enrolled for study in 2009 from a single village in central China. Markers of liver function (alanine aminotransferase (ALT) and aspartate aminotransferase (AST)) and infection (anti-HCV antibodies, CD4⁺ T cell counts, HCV genotype, and HCV viral load) were measured at baseline and follow-up (in July 2012). At follow-up,the subjects were grouped according to ongoing HCV mono-infection (n=129), ongoing HCV/HIV co-infection (n=98), spontaneously resolved (SR)-HCV in mono-infection (n=65), and SR-HCV in HCV/HIV co-infection (n=48) for statistical analysis.</p><p><b>RESULTS</b>Almost all of the subjects in the ongoing HCV mono-infection group showed high levels of HCV antibodies (S/CO more than or equal to 10), but the majority of the subjects in the SR-HCV in mono-infection group and in the ongoing HCV/HIV co-infection group. The SR-HCV mono-infection group showed a remarkable decrease in HCV antibodies from 2009 (HIV:7.75 ± 3.8; HIV+:7.61 ± 3.47) to 2012 (HIV:5.51 ± 3.67; HIW:4.93 ± 3.35) (HIV:t =10.67, P less than 0.01; HIV+:t =9.52, P less than 0.01). The ongoing HCV/HIV co-infection group showed a positive correlation between HCV antibodies S/CO ratio and CD4⁺ T cell count (r=028, P=0.008). In the ongoing HCV mono-infection group,the levels of HCV antibodies were significantly higher in individuals infected with HCV-1b than in those with HCV-2a (14.74 ± 1.68 vs.14.08 ± 1.44, t=2.20, P=0.03). In the ongoing HCV/HIV co-infection group, the numbers of subjects with elevated (more than 40 U/L) liver function markers were significantly different according to the HCV genotype infection:HCV-1b:ALT, 25/42 vs.16/56 (x²=9.45, P=0.002); HCV2a:AST, 28/42 vs.18/56 (x²=11.49, P=0.001). The HCV RNA positive rate was significantly higher in subjects with high HCV antibody cutoff values (S/CO more than or equal to 10) than in those with low HCV antibody (S/CO less than 10) (HIV:128/151 vs.1/43, x²=102.11, P less than 0.01; HIV+:88/98 vs.10/48, x²=69.44, P less than 0.01), regardless of HIV co-infection. Significantly more subjects in the ongoing HCV mono-infection group had elevated (more than 40 U/L) ALT or AST than the subjects in the SR-HCV mono-infection group with high levels of HCV antibody (S/CO more than or equal to 10) (ALT:57/128 vs.2/23, x²=10.52, P=0.001; AST:57/128 vs.0/23, x²=16.45, P less than 0.01).</p><p><b>CONCLUSION</b>Serum HCV antibody levels, in combination with other clinical information such as liver function and HIV infection status, may aid in the preliminarily evaluation of an individual's HCV infection status and likelihood for spontaneous resolution. Low levels of HCV antibody (S/CO less than 10) may indicate a better chance of SR-HCV, after ruling out the possibility of suffering from immunosuppressive diseases such as HIV infection.</p>

The frequency,phenotypes and invtiro cytotoxic effects of icrculating CD 56+T cells in the patients with chornic HCV infection / 中华微生物学和免疫学杂志

Objective To explore the cell frequency , phenotypes and in vitro cytotoxic effects of circulating CD56+T cells in the patients with chronic HCV infection .Methods Peripheral blood mononu-clear cells (PBMCs) were isolated from 33 patients with HCV chronic infection and 21 healthy subjects. Multi-color flow cytometry was used to analyze cell frequency , expressions of activating receptors ( NKG2C, CD16 and NKp46) and inhibitory receptors (NKG2A and CD158a) on CD56+T cells.The functional mark-er for cytotoxic effects (CD107a) on circulating CD56+T cells and their cytokines expression (IFN-γand TNF-α) with or without stimulation of K 562 human Leukemia cell line were also analyzed .Then the correla-tions among the expressing levels of CD 107 a, IFN-γand TNF-αwere investigated .Results The frequency of CD56+T cells in periphery lymphocytes were significantly decreased in the patients with chronic HCV in -fection as compared with that in healthy controls ( P=0.018 ).The expressions of activating receptors (NKG2C, CD16 and NKp46) on CD56+T cells from HCV infected patients were decreased (P=0.015 for NKG2C, P=0.036 for CD16 and P=0.001 for NKp46), while there was no significant change in the ex-pressions of inhibitory receptors (P>0.05 for both CD158a and NKG2A).The concentrations of IFN-γand TNF-αsecreted by CD56+T cells in the patients with chronic HCV infection were significantly decreased with or without K562 stimulation (P0.05). The expressions of CD107a, IFN-γand TNF-αwere closely related under the stimulation of K562(r>0.80, P<0.0001).Conclusion The frequency of CD56+T cells was reduced in patients with chronic HCV infec-tion.Moreover, cytotoxic effects and cytokines production mediated by CD 56+T cells were also significantly impaired, indicating that the dysfunction of circulating CD 56+T cells might be associated with the persist-ence of chronic HCV infection .

Effects of chronic HCV infection and spontaneously resolved HCV infection on serum trace elements and their relationships with serum albumin and circulating HCV RNA load / 中华微生物学和免疫学杂志

Objective To comparatively analyze serum levels of zinc , iron, magnesium, calcium, copper and phosphorus and their relationships with serum albumin and circulating HCV viral load in patients with chronic HCV infection , subjects with spontaneously resolved HCV infection and healthy subjects . Methods Serum levels of the six trace elements in patients with chronic HCV infection (n=59), subjects with spontaneously resolved HCV infection (n=65) and healthy subjects (n=48) were measured by a flame atomic absorption spectrophotometer and then a comparative analysis was performed to analyze the differences among the three groups .The relationships of the six trace elements with serum albumin and HCV viral load were analyzed among patients with chronic HCV infection .Results Compared with healthy subjects , the levels of serum zinc were significantly decreased , but serum levels of iron , copper , phosphorus were signifi-cantly increased in patients with chronic HCV infection .There was a significant positive correlation between the levels of zinc(r=0.4022, P=0.0016)and albumin in patients with chronic HCV infection with ALT no less than 40 IU/L, whereas negative correlations were presented between trace elements of iron ( r=-0.3001, P=0.0209), copper (r=-0.3856, P=0.0036), phosphorus (r=-0.3600, P=0.0075) and serum albumin.The circulating HCV viral load was negatively correlated with serum zinc (r=-0.4367, P=0.0005), but positively correlated with serum copper (r=0.3328, P=0.0139).The serum levels of six trace elements showed no significant differences between healthy subjects and spontaneous resolvers of HCV infection.Moreover, no significant differences of serum calcium and magnesium were found among the three groups.Conclusion Chronic HCV infection can induce abnormal serum levels of zinc , iron, copper and phosphorus and the abnormal serum levels of trace elements were closely related with liver function and HCV viral load.With the spontaneous clearance of HCV infection , the serum levels of trace elements could restore to normal .

Effect of triptolide on sensitivity of K562/A02 cell line to adriamycin / 中国实验血液学杂志

This study was aimed to evaluate the effect of triptolide (TPL) on the reversal of multidrug resistance in K562/A02 cell line. The sensitivity of K562 and K562/A02 to adriamycin (ADM) and reversal of drug resistance were determined with MTT method. The concentration of intracellular ADM and P-glycoprotein expression were detected by flow cytometry. Luciferase reporter gene assay was used to detect the transcriptional activity of MDR1 promoter. The results showed that TPL significantly decreased the resistance degree of K562/A02 cells, inhibited P-glycoprotein expression (mean fluorescent intensity decreased from 123 ± 13 to 39 ± 13) and increased the intracellular concentration of ADM (mean fluorescent intensity increased from 18 ± 5 to 34 ± 6) in K562/A02 cells. Luciferase reporter gene assay demonstrated that TPL inhibited the transcriptional activity of MDR1 promoter by 75%. It is concluded that TPL may effectively reverse the multidrug resistance in K562/A02 cells via modulating P-glycoprotein expression and increasing intracellular ADM accumulation.