Effect of Huanglian Jiedutang on Learning and Memory Ability and Cholinergic System in AD Rats Induced by Aβ1-42 / 中国实验方剂学杂志

Objective:To investigate the effects of Huanglian Jiedutang on learning and memory ability and the cholinergic system in Alzheimer's disease（AD） rats induced by amyloid <italic>β</italic>-protein（A<italic>β</italic>）_1-42. Method:Sixty male SD rats were divided into normal group， model group， huperzine A group （2.1×10^-5 g·kg^-1）， high-， medium- and low dose of Huanglian Jiedutang groups （6，3，1.5 g·kg^-1）. AD rat model was replicated by hippocampal injection of A<italic>β</italic>_1-42. After 4 weeks of treatment， Morris water maze test was performed. Hematoxylineosin （HE） staining was used to observe the pathological changes of rat hippocampus. Sampling blood from abdominal aorta was taken. Acetylcholine （ACh）， acetylcholinesterase （AchE） and choline acetyltransferase （ChAT） in serum and hippocampus were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of hippocampal <italic>α</italic>7 nicotinic acetylcholine receptor （<italic>α</italic>7nAChR） protein was detected by Western blot. The expression of hippocampal <italic>α</italic>7nAChR mRNA was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with the normal group， there were obvious pathological changes in the model group，such as neuron necrosis in the cerebral cortex，pyramidal cell or granular cell necrosis in the hippocampus，disorder of arrangement and inflammatory cell infiltration，prolonged escape latency，decreased escape platform times，decreased residence time in the effective area and swimming path in the effective area （<italic>P<</italic>0.05，<italic>P<</italic>0.01）. The contents of <italic>α</italic>7nAChR mRNA，ACh，AchE，ChAT，<italic>α</italic>7nAChR in the hippocampus decreased （<italic>P<</italic>0.01）. Compared with the model group，the escape latency of the middle dose group was shorter （<italic>P<</italic>0.05）， the escape platform times，the swimming path in the effective area and the residence time in the effective area increased （<italic>P<</italic>0.05，<italic>P<</italic>0.01）， the contents of serum ACh，ChAT， hippocampal AchE，ChAT and <italic>α</italic>7nAChR increased （<italic>P<</italic>0.05，）. The expression of hippocampal <italic>α</italic>7nAChR protein significantly increased （<italic>P<</italic>0.01）， the residence time of effective area in high dose group was prolonged （<italic>P<</italic>0.01）， the times of escape platform increased，and the contents of serum ACh，ChAT and hippocampal ACh，AchE，<italic>α</italic>7nAChR protein and <italic>α</italic>7nAChR mRNA increased （<italic>P<</italic>0.05）. Conclusion:Huanglian Jiedutang can significantly improve the learning and memory ability of AD rats induced by A<italic>β</italic>_1-42，and its mechanism may be related to the improvement of cholinergic system damage and enhancement of cholinergic system function induced by A<italic>β</italic>_1-42.

Tweety-Homolog 1 Facilitates Pain via Enhancement of Nociceptor Excitability and Spinal Synaptic Transmission / 神经科学通报·英文版

Tweety-homolog 1 (Ttyh1) is expressed in neural tissue and has been implicated in the generation of several brain diseases. However, its functional significance in pain processing is not understood. By disrupting the gene encoding Ttyh1, we found a loss of Ttyh1 in nociceptors and their central terminals in Ttyh1-deficient mice, along with a reduction in nociceptor excitability and synaptic transmission at identified synapses between nociceptors and spinal neurons projecting to the periaqueductal grey (PAG) in the basal state. More importantly, the peripheral inflammation-evoked nociceptor hyperexcitability and spinal synaptic potentiation recorded in spinal-PAG projection neurons were compromised in Ttyh1-deficient mice. Analysis of the paired-pulse ratio and miniature excitatory postsynaptic currents indicated a role of presynaptic Ttyh1 from spinal nociceptor terminals in the regulation of neurotransmitter release. Interfering with Ttyh1 specifically in nociceptors produces a comparable pain relief. Thus, in this study we demonstrated that Ttyh1 is a critical determinant of acute nociception and pain sensitization caused by peripheral inflammation.

Anti-tumor Effect of Astragaloside by Inducing M1 Macrophage Polarization / 中国实验方剂学杂志

Objective:To investigate the effect of astragaloside on the macrophage polarization and the possible anti-tumor immunity mechanism of astragaloside. Method:The cytotoxic effect of different concentrations of astragaloside at different time points on macrophage was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT), in order to choose the suitable concentration of astragaloside, macrophages were co-cultured with tumor cells at the ratio 1:1, and the effect of astragaloside on macrophage-mediated lysis of tumor cells was performed by biophotonic cytotoxicity assay after the mixed cells were effected with 0.1 mg·L-1 astragaloside for 24 h. Macrophages were dealt with 0.1 mg·L-1 astragaloside for 24h, the expressions of CD16/32 and CD206 in macrophages were performed by flow cytometry, the mRNA expressions of macrophage inducible nitric oxide synthase (iNOS), Arginine-1 (Arg-1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-12 (IL-12), interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) were measured by Real-time PCR, the protein expressions of macrophage signal transducers and activators of transcription 1 (STAT1) and phosphorylation signal transducers and activators of transcription 1 (p-STAT1) were determined by Western blot. Result:Astragaloside had no effect on the viability of macrophages with 0.1 mg·L-1. Compared with control group, astragaloside obviously enhanced the macrophage-mediated lysis of tumor cells according to the biophotonic cytotoxicity assay, induced the M1 macrophage marker CD16/32 expression according to flow cytometry, increased the mRNA expressions of iNOS, IL-1β, TNF-α and IL-12 according to the Real-time PCR, and promoted the phosphorylation of STAT1 in macrophages on the basis of Western blot. Conclusion:Astragaloside could induce M1 macrophage polarization by increasing the phosphorylation of STAT1, and initiate macrophage-related anti-tumor immunity response.

Proliferation-inhibiting and multidrug-resistant reversing effect of bortezomib on human HL-60 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the proliferation-inhibiting and multidrug-resistant reversing effect of bortezomib on human HL-60 cells, and to explore the mechanism of bortezomib-induced proliferation inhibition in human leukemia cells.</p><p><b>METHODS</b>The multidrug resistant leukemia cell lines HL-60/DNR and HL-60/VCR cells were used as models, and sensitive HL-60 cells as a control. The cytotoxicity of bortezomib on HL-60, HL-60/DNR, HL-60/VCR cells were measured by MTT method, and the non-cytotoxicity dose was determined as reversible dose. The cells were divided into 4 experimental groups: HL-60/DNR + DNR, HL-60/DNR + DNR + bortezomib, HL-60/VCR + VCR, HL-60/VCR + VCR + bortezomib. The bortezomib resistant reversal fold was calculated. The levels of XIAP, cIAP-1, and cIAP-2 mRNA and proteins expression and the activation of NF-κB of the HL-60/DNR, HL-60/VCR cells were examined by quantitative real time RT-PCR and western blot respectively after treated with gradually increasing concentrations of bortezomib (10, 40, 80 nmol/L) for 48 hours.</p><p><b>RESULTS</b>Bortezomib inhibited the cell growth of HL-60, HL-60/DNR, and HL-60/VCR in a concentration-dependent manner. The IC(50) values were (28.90 ± 3.99), (81.19 ± 9.34), and (73.48 ± 8.94) nmol/L, respectively. After treated with 10nmol/L bortezomib for 48 hours, the IC(50) value of DNR to HL-60/DNR decreased from (12.90 ± 1.75) µmol/L to (3.54 ± 0.57) µmol/L (P < 0.01), and that of VCR to HL-60/VCR from (33.25 ± 7.28) µmol/L to (9.97 ± 1.15) µmol/L (P < 0.01). The reversal fold (RF) values were 3.32 ± 0.53 and 2.64 ± 0.28, respectively. Bortezomib down-regulated the levels of XIAP, cIAP-1, and cIAP-2 mRNA and protein expression and inhibited the NF-κB activation in a concentration-dependent manner.</p><p><b>CONCLUSION</b>Bortezomib can inhibit the proliferation of HL-60 cells and reverse multidrug-resistance in the cells. The possible mechanism is associated with down-regulation of IAPs expression.</p>

Expression of TEKT4 protein decreases in the ejaculated spermatozoa of idiopathic asthenozoospermic men / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia.</p><p><b>METHODS</b>We separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot.</p><p><b>RESULTS</b>RT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 +/- 0.13 vs 0.75 +/- 0.15, t = 4.325, P < 0.05), and Western blot confirmed the results of RT-PCR (0.48 +/- 0.14 vs 0.69 +/- 0.13, t = 5.939, P < 0.05).</p><p><b>CONCLUSION</b>The expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.</p>

Expression of Kindlins and angiopoietins in acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to explore the expression of Ang-1, Ang-2, Tie-2, Kindlin-2, Kindlin-3 in different leukemia cell lines and bone marrow of acute myeloid leukemia (AML) patients and its clinical significance. The levels of Ang-1, Ang-2, Tie-2, Kindlin-2, Kindlin-3 in bone marrow of AML patients and nontumorous control group as well as leukemia cell lines (K562, KG-1a, U937, HL-60 and Jurkat) were detected by RQ-PCR. The difference of positive rate and expression level between AML patients and controls was analyzed. The relation between 5 genes and their relationship with typing and prognosis of AML were investigated. The results showed that Ang-1, Ang-2, Kindlin-3 expressed in K562, KG-1a, U937, HL-60 and Jurkat cells. Tie-2 only expressed in KG-1a and HL-60 cells. Kindlin-2 expressed in K562, KG-1a and HL-60 cells. All of these 5 genes expressed in AML patients and nontumorous controls. The expression level of Ang-1 and Ang-2 in patients with higher WBC count (≥ 30 × 10(9)/L) was significantly higher than that in patients with lower WBC (< 30 × 10(9)/L, P < 0.001, P < 0.001). The expression level of Ang-1 and Ang-2 in patients with t(8;21) or t(15;17) was significantly lower (P < 0.001, P = 0.005). In the NCCN better-risk group, Ang-1 expressed lower (P = 0.020). The group with lower expression of Ang-1 showed a higher complete remission (CR) rate (P = 0.027). The expression level of Kindlin-2 was lower in AML patients (P = 0.010), lower in patients with higher WBC (≥ 30 × 10(9)/L, P = 0.020), and higher in patients with t(8;21) or t(15;17). The expression levels of both Kindlin-2 and Kindlin-3 were significantly higher after CR (P < 0.001, P = 0.004). It is concluded that Ang-1 closely correlated with the poor prognosis of AML. Kindlin-2 lowly expresses in AML, which has a close relation with the favorable prognosis of AML. Kindlin-2 can be a marker for favorable prognosis of AML.

Therapeutic effect of endovascular treatment on patients with epileptic attack induced by carotid artery or vertebral artery stenosis / 中华神经医学杂志

Objective To observe the therapeutic effect of endovascular treatment on patients with epileptic attack induced by carotid artery or vertebral artery stenosis.Methods Two epileptic attack patients,diagnosed as having symptomatic carotid artery or vertebral artery stenosis by DSA,were performed endovascular stenting and balloon dilation.Case one had a serious artery stenosis at the left original internal carotid artery and moderate artery stenosis at the right proximal internal carotid artery and vertebral artery.Case two had 1 occlusion at the right original intemal carotid artery and serious artery stenosis at the clinoid segment of left internal carotid artery and the double proximal vertebral arteries restenosis after percutaneous transluminal angioplasty and stenting (PTAS) for symptomatic ostial vertebral artery stenosis.Two patients were treated with PTAS and balloon angioplasty.Follow up was performed for one to three years.Results The clinical symptoms of these two patients disappeared aftter the operation.Case one was followed up for almost 3 years and case two for more than one year; no relapse was noted for their regular taking platelet aggregation inhibitor and without taking antiepileptic drug.Conclusion Endovascular treatment is a safe and efficient method for treating patients with epileptic attack induced by carotid artery or vertebra-basilar artery stenosis.

CatSper1 protein and idiopathic asthenozoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role of the cation channel of sperm 1 (CatSper1) protein in the pathogenesis of idiopathic asthenozoospermia.</p><p><b>METHODS</b>Sperm samples from patients with idiopathic asthenozoospermia were separated by Percoll discontinuous density gradients, and the distribution and expression of the CatSper1 protein were determined by immunocytochemistry. Western blotting was used to detect the different expressions of CatSper1 in the ejaculated sperm from the normal control, mild asthenozoospermia, moderate asthenozoospermia and severe asthenozoospermia groups, followed by statistical analyses.</p><p><b>RESULTS</b>The expression of CatSper1, located in the principle piece of the sperm tail, was reduced significantly in the samples from the idiopathic asthenozoospermia patients as compared with the normal controls (t = 2.188, P = 0.042). The relative contents of the CatSper1 protein in the sperm of the control, mild asthenozoospermia, moderate asthenozoospermia and severe asthenozoospermia groups were 0.806 +/- 0.266, 0.669 +/- 0.207, 0.505 +/- 0.214 and 0.295 +/- 0.162, respectively, significantly decreased in the asthenozoospermia patients in comparison with the normal controls (P <0.05). There was a positive correlation between the percentage of progressively motile sperm and the relative content of the CatSper1 protein (r = 0.633, P = 0.000).</p><p><b>CONCLUSION</b>The decreased or abnormal expression of the CatSper1 protein may be a factor involved in the pathogenesis of idiopathic asthenozoospermia.</p>

Relationship between levels of serum IL-1β and IL-1Ra and restenosis in patients with intra and extra-cranial arteriostenosis after stent-assisted angioplasty / 中华神经医学杂志

Objective To observe the levels of serum interleukin-1β(1L-1β)and interleukin-1 receptor antagonist(IL-1Ra),the IL-1Ra/IL-1β ratio and the relationship between the levels of thern and the restenosis in patients with intra-and extra-cranial arteriostenosis after stent-assisted angioplasty.Methods Thirty-one patients with cerebral artery stenosis,admitted to our hospital from April 2003 to March 2006,were treated with stent-assistant angioplasty and followed up for 6-12 months.The relationship was analyzed between the restenosis of cerebral artery and both the levels of serum IL-1β,IL-1Ra and the IL-1 Ra/IL-1β ratio before and 1 h,1,3 and 5 d after stent-assisted angioplasty.Results The condition of 31 stents in 31 blood vessels of cerebral arteries was observed and followed up 6 to 12 months after the operation.Restenosis was noted in 6(19.3%)with 3 restenosis superior to 50% and 3 restenosis from 10%-30%.No obvious difference of the levels of serum IL-1β and IL-1Ra,the IL-1Ra/IL-1β ratio between restenosis group and non-restenosis group before the operation was found (P>0.05);IL-11β and IL-1Ra levels were positively correlated before the operation.The levels of serum IL-1β and IL-1Ra 1h,1,3 and 5 d after the operation were superior to those before the operation,respectively(P<0.05).The levels of serum IL-1β and IL-1Ra in the restenosis group showed no significant difference to those in the non-restenosis group 6 to 12 months alter the operation(P>0.05);however,the IL-1Ra/IL-1β ratio in the restenosis group was significantly lower than that in the non-restenosis group (P<0.05);positive correlation between the levels of IL-1β and IL-1Ra was observed.Conclusion The levels of serum IL-1β and IL-1Ra were associated with the inflammatory process after the operation and the immune maladjustment of IL-1β and IL-1Ra might be correlative to the restenosis,indicating that the IL-1Ra/IL-1β ratio might be an available index for monitoring the restenosis.

Stent fracture and restenosis after percutaneous transluminal angioplasty and stenting for symptomatic ostial vertebral/subclavian artery stenosis: a report of 3 cases and literature review / 中华神经医学杂志

Objective To discuss mechanism and control measures of stent fracture and restenosis after percutaneous transluminal angioplasty and stenting (PTAS) for symptomatic ostial vertebral/subclavian artery stenosis. Methods A retrospective analysis was performed on 3 patients with stent fracture after receiving PTAS for symptomatic ostial vertebral/subclavian artery stenosis.Simple radiographic, ultrasonographic and clinical follow-up examinations were estimated. Related articles on coronary stent fracture were gone over, consulting in the types, cumulative incidence and occurrence time of adverse events, risk factors and preventive measures. Results Stent fractures of 3 patients with symptomatic ostial vertebral /subclavian artery stenosis were associated with in-stent restenosis and occlusion. Two of the 3 patients treated with the balloon angioplasty and after balloon dilatation, and the patients exhibited relief of symptoms. One patient was only managed for vascular disease risk factors, and no developing recurrent symptoms were noted during the follow-up period.Conclusions Stent fracture might appear in patients performed PTAS for symptomatic ostial vertebral /subclavian artery stenosis, and regular check is needed. Individual treatment was emphasized in case of serious symptoms appeared.

CT perfusion imaging study of the hemodynamics in patients with intracerebral hemorrhage / 中华神经医学杂志

Objective To investigate the hemodynamic changes in patients with intracerebral hemorrhage (ICH) using CT perfusion (CTP) imaging and the relation between cerebral perfusion and the Scandinavian Stroke Scale (SSS) of the patients. Methods Forty-one patients with supratentorial ICH underwent plain CT scanning and CTP imaging at different times after ICH onset (from 5 h to 14 days). The impact of the time after ICH onset, hematoma volume, edema area, mean hlood pressure (MBP) and blood glucose on the hemodynamics of the patients was assessed, and the correlation between the SSS and the hemodynamic changes evaluated. Resnlts The cerebral blood flow in the edema area (CBF1), the peripheral area of the edema (CBF2, within 1 cm to the edema) and the distant cortical area from the hematoma (CBF3) showed significant differences (r=0.027, P=0.870) and fluctuated with time. CBF1 showed a positive linear relation with CBF2 (r=0.334, P=0.035), but neither of them was correlated to CBF3 (r=0.027, P=0.870;r=0.142, P=0.383). CBF1 also showed positive linear relations with relative cerebral blood volume (rCBV1) (r=0.803,P=0.000) and the peak time (PT1) (r=-0.52, P=0.752). The reduction of CBF1 was inversely correlated to the hematoma volume and the edema area (r=-0.501, P=0.001;r=-0.589, P=0.000), but not related with MBP or blood glucose (r=0.141, P=0.386;r=0.014, P=0.930). The area of ischemic injury (the ischemic area defined by CBF parameters-the hematoma area, edema area (r=0.449, P=0.003;r=0.645, P=0.000), but not to the MBP or blood glucose (r=-0.047, P=0.769;r=0.141,P=0.378). SSS was found to correlate to the volume and location of the hematoma and the reduction of CBF and CBV (r=-0.418, P=0.007;r=0.542, P=0.000;r=0.376, P=0.017;r=0.312, P=0.050), but not to the ischemic and edema area (r=-0.283, P=0.073;r=-0.163, P=0.308). Conclusion CBF is reduced in the edema area, peripheral area of the edema and the distant cortical area from the hematoma but showing different patterns of variation. Ischemic injury is present in the edema area around the hemotema, and its scope and severity is related with the hemotoma volume and the size of the edema area and may vary with time. The location and volume of the hematoma as well as the perfusion level in the edema area all affect the SSS of the patients.

Univariate analysis of collateral circulation and nerve function in patients with severe stenosis or occlusion of internal carotid artery / 中华神经医学杂志

Objective To investigate the relation between compensation of collateral circulation and the severity of clinical neurologic deficit in patients with severe stenosis or occlusion of the internal carotid artery (ICA). Methods National Institutes of Health Stroke Scale (NIHSS) examination was performed in 52 patients with cerebral infarction caused by consecutive stenosis or occlusion of ICA during admission and digital subtraction angiography (DSA) was employed to evaluate the compensation of collateral circulation to analyze the results through statistical methods. Results DSA demonstrated occlusion oflCA in 18 patients, severe stenosis of unilateral ICA in 28, and severe stenosis of bilateral ICA in 6. Anterior communicating artery (ACoA) compensation was presented in 18 patients;posterior communicating artery (PCoA) compensation appeared in 8;both ACoA and PCoA compensations were showed in 12. Only 14 patients have no compensation by Willis circle. Significant differences of average ranks of the NIHSS were found in non Willis circle (35.75) and Willis circle (23.09) (P<0.05);while no significant differences were noted in ACoA (12.42) and PCoA (15.94) compensations (P>0.05). Conclusions Effective collateral circulation can be established spontaneously through multiple ways when occlusion or severe stenosis of ICA appears. The NIHSS scores and the compensation, the location of vessel occlusion are significantly correlative. It is extremely important to investigate the collateral circulation carefully through DSA before treatment intervention.

Protective effect of hirudo extract liquid against toxic injury of astrocytes induced by thrombin in vitro / 中华神经医学杂志

Objective To study the cell toxicity of thrombin in astrocytes in vitro and the protective effect of hirudo extract liquid (HEL) on the injured astrocytes. Methods Astrocytes were isolated from Wistar rats' cerebral cortex and cultured in vitro, and observed under a phase contrast microscope for growth status. Cell activity was measured with MTT assay. The survival of astrocytes was investigated after exposed to a selected concentration of thrombin ranging from 0.1 to 100 U/mL or to HEL ranging from 0.25 to 4 mg/μL by observing cell morphology under an inverted phase-contrast microscope and measuring the lactate dehydrogenase (LDH) activity (a marker of cell death) in cell supernatant. Expressions of HSP70 and TGFβ-1 protein in astrocytes were investigated by immunohistochemistry. Results (1) Thrombin (1-100 U/mL) had toxicity on astrocytes in vitro in a dose-dependent manner (F=118.65, P=0.000). (2) HEL (0.25-4 mg/μL) could significantly reduce the cell toxicity of 10 U/mL thrombin in astrocytes (F=156.08, P=0.000). With the increasing concentration of HEL, the protection of HEL was accordingly enhanced, and it even increased the expressions of HSP70and TGFβ-1. Conclusions HEL could accelerate the proliferation of astrocytes, enhance the expressions of HSP70 and TGFβ-1 protein, so as to significantly depress the cell toxicity of thrombin to astrocytes.

Relationship between cortical watershed infarction and carotid artery stenosis and a follow-up and control study on prognosis after stent insertion / 中华神经医学杂志

Objective To investigate the relationship between cortical watershed infarction and carotid artery stenosis and evaluate the stent insertion operation.Methods After 23 cortical watershed infarction patients diagnosed by CT or MRI received DSA detection,we performed stent insertion operationon 11 patients according to their requirements,and conservative treatment on the remaining 12 patients.All the patients underwent follow up for 6-12 months post-operatively.Results Among the 23 cortical watershed infarction patients,22 Were detected with carotid artery stenosis.Statistical analysis showed that the degree of carotid artery stenosis was associated With the elinical svmDtoms and the volume of steal phenomenon(P<0.05);further,the artery stenosis improvement was over 90%with the stent inserted;conversely,dizziness and steal phenomenon disappeared.The post procedure follow-up,ranging 6-12 months,showed that the patients with stent insertion got less new symptoms,steal phenomenon and artery stenosis,compared with the patients with conservation treatment(P<0.05).Conclusion Cortical watershed infarction is associated with carotid artery stenosis.The stent insertion iS useful for the treatment ofcarotid artery stenosis and prevention of cortical watershed infarction.

Antigenicity of hepatitis C virus F protein and serum prevalence of anti-F in HCV-infected patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To examine the antigenicity of hepatitis C virus (HCV) F protein and investigate serum prevalence of anti-F in HCV-infected patients.</p><p><b>METHODS</b>Eleven pairs of overlapping primers were used to synthesize the full-length HCV f gene, from which the truncated HCV f65 gene fragment was amplified by PCR. HCV f65 gene was then cloned into pET32a(+), and transformed into E. coli strain Plyss (DE3). This recombinant E.coli was induced by IPTG for the production of HCV F65 protein. The expressed HCV F65 protein, purified by Ni-NTA agarose, was further used in ELISA to detect serum anti-F, and to immunize rabbits for making polyclonal anti-F. The rabbit polyclonal anti-F was purified by Staphylococcus aureus protein A agarose.</p><p><b>RESULTS</b>After recombinant pET32a(+)-f65 was constructed successfully, HCV F65 protein was expressed and purified. The purified HCV F65 protein was used as a capture antigen in ELISA to detect serum anti-F in HCV infected patients (n = 30). The result showed that the mean A450 value and the positive rate of serum anti-F were 0.125+/-0.061 and 63.3%, respectively. The rabbit-derived polyclonal anti-F reacted specifically with HCV F65 protein, of which the titer was 1:30,000.</p><p><b>CONCLUSION</b>Our expressed HCV F65 protein is of antigenicity, and can be used to determine serum anti-F. Anti-F IgG does exist in the sera of the HCV-infected patients. Moreover, the rabbit-derived polyclonal anti-F can be used to detect HCV F protein.</p>

Study of the therapeutic effects of free radical scavenger edaravone on cerebral hemorrhage in rats / 中华神经科杂志

Objective To explore therapeutic effects and mechanisms of radical scavenger edaravone on experimental cerebral hemorrhage.Methods Two hundred-forty male SD rats were divided randomly into four groups:control group,cerebral hemorrhage group,edaravone treatment group before operation (A) and edaravone treatment group after operation (B).Experimental cerebral hemorrhage model was made according to the method reported by Rosenberg.Water quantity contained in brain and nervous missing sign were observed,meanwhile the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in brain tissue were measured.Results Compared with cerebral hemorrhage group,nervous missing sign and water quantity contained in brain obviously changed in edaravone treatment group (P