Dynamic changes in cytoskeletal elements following acute cerebral ischemia and reperfusion in rats / 中国药理学通报

Aim To investigate the dynamic time-course changes in neuronal cytoskeleton after acute ischemia and reperfusion in rats. Methods Reperfusion was performedin rats by blocking the middle cerebralarteryfor 90 min, then therats wereobserved and collected at different time points. The brain damage wasobserved by Nissl staining,and neurobehavioural function was evaluated with neurological deficit score and forelimb placement test. The cellular changes in the alternations of cytoskeletal elements including microtubule associated protein 2 (MAP2) and neurofilament heavy chain (NF-H) were observed by immunohistochemistry staining and Western blot. Impaired axons, dendrites and cytoskeletal alternations were detected by electron microscope. Results Brain damage and neurobehavioural function were gradually aggravated with the prolongation of reperfusion. Brain damage appeared earlier and more severe in striatum than in cortex. Moreover, decreased MAP2-related and increased NF-H-related immunoreactive intensities were found in the ischemic areas. Impaired cytoskeletal arrangement and reduced dense were indicated. Damaged cytoskeletal components such as microtubules and neurofilament arrangement, decreased axonal filament density, and swelled dendrites were observed after cerebral ischemia reperfusion by ultrastructural observations. Conclusions Different brain regions have diverse tolerance to ischemia-reperfusion injury. Major elements of neuronal cytoskeleton show dynamic responses to ischemia and reperfusion, which may further contribute to brain damage and neurological impairment following MCAO and reperfusion.

Research progress on hypoxia/reoxygenation model of nerve cells in vitro / 中国药理学通报

Ischemic stroke is the second leading cause of human death and the third reason of disability. Meanwhile, the incidence is rising year after year worldwide. Ischemic stroke could cause ischemia-reperfusion injury after blood recanalization treat-ment, but the mechanism of ischemia-reperfusion injury is still not very clear, so it is necessary to build a preclinical model with specific characteristics. Up to now, animal experiments have been still complicated, and the culture of brain slices has some limitations. The cell model in vitro has become a simplified and valuable tool widely used by researchers. The paper systematically summarizes the common type of nerve cells, and further analyzes establishment methods and principle, relevant research progress on the in vitro model of ischemia-reperfusion, in order to provide reference for rationally selecting hypoxia and reoxygenation model for basic research on cerebral ischemia and reperfusion and drug screening.

Research on cellular damages and astrocyte activation after cerebral ischemia and reperfusion / 中国药理学通报

Aim To observe cellular damage and astrocyte activation at different time points of cerebral ischemia and reperfusion. Methods The middle cerebral artery of male SpragueDawley rats was occluded for 90 min followed by different time points of reperfusion. Eighty-five SPF male SD rats were randomly divided into control group (Sham), IR3, 6, 12, 24 and IR48h (MCAO followed by 48 h of reperfusion) group. Cerebral ischemia and reperfusion injury was observed by HE staining, and the structure of astrocytes was estimated with transmission electron microscopy (TEM). GFAP expression was detected by immunofluorescence staining and Western blot. Results Cerebral ischemia following by different time points of reperfusion led to different degrees of cellular damage, which was the most serious at 24 h of reperfusion. TEM showed destruction of astrocytes structure, swollen organelles and broken mitochondrial ridge. After cerebral ischemia-reperfusion, the expression levels of GFAP were significant up-regulated in the ischemic penumbra cortex and the highest was at 48 h of reperfusion, indicating astrocytes were activated. In addition, the results showed the gradual decrease in GFAP expression in the infarct core. Conclusions After cerebral ischemia-reperfusion, cellular damage is aggravated, and astrocytes are gradually activated in the ischemic penumbra. With the extension of reperfusion time, the boundaries of infarct area and ischemic area are gradually clear, and scarring may occur.

Mechanism of neuronal death in ischemic stroke / 中国药理学通报

Stroke is the second leading cause of death in the world, of which about 60 % - 80 % are ischemic stroke. Ischemic stroke will inevitably cause the damage of neurons in the core area. With the increase of ischemic time, other neurons in the ischemic penumbra will also die due to the loss of " signal connection", and further lead to body dysfunction. In view of the complexity of neuronal death mechanism after ischemic stroke, understanding the action principle of death mechanism can better save ischemic penumbra neurons. This review mainly expounds several main mechanisms and potential therapeutic targets of neuronal death after ischemic stroke, so as to provide basis and help for the improvement of action mechanism research and drug development.

Effect of Xiaoxuming Decoction on synaptic plasticity following acute cerebral ischemia-reperfusion in rats / 中国中药杂志

This study aims to explore the effect of Xiaoxuming Decoction on synaptic plasticity in rats with acute cerebral ischemia-reperfusion. A rat model of cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion(MCAO). Rats were randomly assigned into a sham group, a MCAO group, and a Xiaoxuming Decoction(60 g·kg~(-1)·d~(-1)) group. The Longa score was rated to assess the neurological function of rats with cerebral ischemia for 1.5 h and reperfusion for 24 h. The 2,3,5-triphenyltetrazolium chloride(TTC) staining and hematoxylin-eosin(HE) staining were employed to observe the cerebral infarction and the pathological changes of brain tissue after cerebral ischemia, respectively. Transmission electron microscopy was employed to detect the structural changes of neurons and synapses in the ischemic penumbra, and immunofluorescence, Western blot to determine the expression of synaptophysin(SYN), neuronal nuclei(NEUN), and postsynaptic density 95(PSD95) in the ischemic penumbra. The experimental results showed that the modeling increased the Longa score and led to cerebral infarction after 24 h of ischemia-reperfusion. Compared with the model group, Xiaoxuming Decoction intervention significantly decreased the Longa score and reduced the formation of cerebral infarction area. The modeling led to the shrinking and vacuolar changes of nuclei in the brain tissue, disordered cell arrangement, and severe cortical ischemia-reperfusion injury, while the pathological damage in the Xiaoxuming Decoction group was mild. The modeling blurred the synaptic boundaries and broadened the synaptic gap, while such changes were recovered in the Xiaoxuming Decoction group. The modeling decreased the fluorescence intensity of NEUN and SYN, while the intensity in Xiaoxuming Decoction group was significantly higher than that in the model group. The expression of SYN and PSD95 in the ischemic penumbra was down-regulated in the model group, while such down-regulation can be alleviated by Xiaoxuming Decoction. In summary, Xiaoxuming Decoction may improve the synaptic plasticity of ischemic penumbra during acute cerebral ischemia-reperfusion by up-regulating the expression of SYN and PSD95.

Effect of Xiaoxuming Decoction on activation of astrocytes in acute cerebral ischemia/reperfusion injury / 中国中药杂志

This study investigated the effect of Xiaoxuming Decoction(XXMD) on the activation of astrocytes after cerebral ischemia/reperfusion(I/R) injury. The model of cerebral IR injury was established using the middle cerebral artery occlusion method. Fluorocitrate(FC), an inhibitor of astrocyte activation, was applied to inhibit astrocyte activation. Rats were randomly divided into a sham group, a model group, a XXMD group, a XXMD+FC group, and a XXMD+Vehicle group. Neurobehavioral changes at 24 hours after cerebral IR injury, cerebral infarction, histopathological changes observed through HE staining, submicroscopic structure of astrocytes observed through transmission electron microscopy, fluorescence intensity of glial fibrillary acidic protein(GFAP) and thrombospondin 1(TSP1) measured through immunofluorescence, and expression of GFAP and TSP1 in brain tissue measured through Western blot were evaluated in rats from each group. The experimental results showed that neurobehavioral scores and cerebral infarct area significantly increased in the model group. The XXMD group, the XXMD+FC group, and the XXMD+Vehicle group all alleviated neurobehavioral changes in rats. The pathological changes in the brain were evident in the model group, while the XXMD group, the XXMD+FC group, and the XXMD+Vehicle group exhibited milder cerebral IR injury in rats. The submicroscopic structure of astrocytes in the model group showed significant swelling, whereas the XXMD group, the XXMD+FC group, and XXMD+Vehicle group protected the submicroscopic structure of astrocytes. The fluorescence intensity and protein expression of GFAP and TSP1 increased in the model group compared with those in the sham group. However, the XXMD group, the XXMD+FC group, and XXMD+Vehicle group all down-regulated the expression of GFAP and TSP1. The combination of XXMD and FC showed a more pronounced effect. These results indicate that XXMD can improve cerebral IR injury, possibly by inhibiting astrocyte activation and down-regulating the expression of GFAP and TSP1.

Therapeutic effect of endoscopic resection for large gastric stromal tumors / 中华消化内镜杂志

Objective To evaluate the feasibility and safety of endoscopic treatment for gastric stromal tumors with maximum diameter of 3 to 5 cm. Methods From April 2010 to April 2016, a retrospective analysis was performed on the data of patients with gastric stromal tumors undergoing endoscopic (29 cases)or laparoscopic(26 cases)resection in the First Affiliated Hospital of Soochow University. Baseline data, perioperative data and follow-up data were collected and analyzed. Results There was significant difference on tumor location between the two groups(χ2=12.173, P=0.007). Tumors mainly located at gastric fundus in the endoscopic group(65.5%,19/29),while at gastric body in the laparoscopic group(61.5%, 16/26). Compared with the laparoscopic group, patients in the endoscopic group had shorter operation time[45(35, 60)min VS 70(60, 85)min, U = 686.000, P<0.05], lesser intraoperative blood loss[15(10,15)mL VS 20(10, 20)mL, U=513.000, P=0.017], and earlier recovery time of gastrointestinal function[12(6, 24)h VS 20(18, 24)h, U=585.500, P<0.001]. Compared with the laparoscopic group, the patients in the endoscopic group had a higher complication rate[55.2%(16/29)VS 11.5%(3/26), χ2=11.543, P<0.001]and a lower intact tumor removal rate [89.7%(26/29)VS 100.0%(26/26),χ2=23.989,P<0.001]. The other perioperative parameters such as the incidence of intraoperative major bleeding, postoperative peritonitis, postoperative fasting time, hospitalization time and total hospitalization expenses showed no statistical different(all P>0.05). The postoperative follow-up time was 30.0(17.5,50.0)and 38.5(26.0,49.8)months in the endoscopic and laparoscopic group,respectively. There was no significant difference in the recurrence rate between the two group[3.4%(1/29)VS 7.7%(2/26),χ2=0.009,P=0.922]. Conclusion Endoscopic resection may be an alternative therapeutic approach for large gastric stromal tumors with shorter operation time, less intraoperative blood loss,and earlier recovery time of gastrointestinal function.

Corneal nerve fiber damage in different stage of diabetic retinopathy patients with type 2 diabetes / 中华眼底病杂志

Objective To observe the corneal nerve fibres damage in different stage of diabetic retinopathy (DR) with type 2 diabetes.Methods A cross-sectional study.One hundred and twenty eyes of 120 patients with type 2 diabetes served as diabetes group.According to International Clinical Diabetic Retinopathy Disease Severity Scales (2002),diabetes patients were classified into 4 subgroups:patients without diabetic retinopathy (NDR),patients with mild or moderate non-proliferative diabetic retinopathy (mNPDR),patients with severe non-proliferative diabetic retinopathy (sNPDR) and patients with proliferative diabetic retinopathy (PDR),each subgroup has 30 eyes of 30 patients.Another 30 eyes of 30 healthy participants served as control group.All eyes were scanned with HRT3 in vivo corneal confocal microscopy.Images of sub-basal nerve plexus were quantified including nerve fiber length (NFL),nerve fiber density (NFD),nerve fiber branch density (NFB),and nerve tortuosity (NT).The correlations of corneal nerve fiber with age,duration of diabetes and glycated hemoglobin (HbA1 c) were analyzed using Spearman correlation analysis.Results NFL,NFD and NFB were found to be significantly lower in diabetic patients (F=147.315,142.586,65.898;P=0.000,0.000,0.000),NT was significantly greater in diabetic patients (F=39.431,P=0.000),when compared to control group.In diabetic patients,NFL,NFD and NFB were gradually reduced with DR severity,NT was gradually increased with DR severity.While the difference ofNFL,NFD,NFB,NT was not statistically significant between sNPDR and PDR subgroups (P＞.0.05),but was statistically significant between other subgroups (P＜0.05).Spearman correlation analysis results showed that age (r=-0.071,-0.080,0.001,0.100;P=0.391,0.328,0.991,0.224) and HbAlc (r=-0.109,-0.115,-0.126,0.025;P=0.238,0.211,0.169,0.781) had no correlation with NFL,NFD,NFB,NT.Duration of diabetes was negatively correlated with the NFL,NFD (r=-0.212,-0.264;P=0.020,0.004),positive correlated with NT (r=0.261,P=0.004),and had no correlation with NFB (r=-0.119,P=0.194).Conclusions Corneal nerve fiber loss and nerve tortuosity increased were found in patients with type 2 diabetes,and even without diabetic retinopathy.The progress of corneal neuropathy was correlated with the severity of DR,but it was not change significantly between sNPDR and PDR.

Simultaneous determination of four constituents in Hydnocarpus anthelmintica by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Hydnocarpus anthelmintica Pierre.METHODS The analysis of 60％ ethanol extract from H.anthelmintica was carried out on a 35 ℃ thermostatic Diamonsil C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 80％ acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 347 nm.RESULTS Luteolin,hydnocarpusol,chrysoeriol and hydnocarpin showed good linear relationships within the ranges of 0.0523 6-1.047 μg (r =0.999 9),0.011 24-0.224 8 μg (r =0.999 9),0.029 46-0.589 2 μg (r =0.999 9) and 0.130 5-2.609 μg (r =0.999 9),whose average recoveries were 102.45％ (RSD=1.9％),98.66％ (RSD=1.8％),97.60％ (RSD=1.6％) and 97.88％ (RSD=1.4％),respectively.CONCLUSION This simple,accurate and sensitive method can be used for the quality control of H.anthelmintica.

Evaluation and nursing of continuous blood purification therapy on acute respiratory distress syndrome in children / 中华实用儿科临床杂志

Objective To investigate the efficacy of continuous blood purification (CBP) in treatment of acute respiratory syndrome (ARDS) in children,and the methods as well as the key points in nursing care.Methods There were 97 cases of ARDS,and 18 cases were treated with CBP.The model for CBP was continuous veno-venous hemofiltration dialysis (CVVHDF) or high volume hemofiltration (HVHF).The observational index included arterial partial pressure of oxygen [pa(O2)],arterial partial pressure of carbon dioxide [Pa(CO2)],mechanical ventilation parameters,complications and side reaction during CBP.Results The pediatric critical illness score in CBP treatment group and non-CBP treatment group were (62.5 ± 25.2) and (78.1 ± 16.3) respectively,and the difference between the 2 groups was significant (P ＜ 0.05) ; the median of pediatric risk of score mortality Ⅲ score in CBP treatment group and non-CBP treatment group were 16(10-20) and 12(7-14),and the difference was significant between the 2 groups (all P ＜0.05).The average duration of CBP in 18 cases was 58 hours(12-232 hours),death occurred in 5 cases,and 2 cases of them were due to giving up treatment,the mortality was 27.8％ ;while in non-CBP treatment group,21 cases occurred death and the mortality was 26.6％.The difference of mortality between the 2 groups was not significant (P ＞ 0.05).Pa (O2)/fraction of inspired oxygen (FiO2) and dynamic compliance (Cdyn) were improved after 2-6 hours CBP,and the pulmonary exudation alleviated after 2 hours treatment.Mechanical ventilation parameters including FiO2,peak inspiratory pressure and positive end expiratory pressure were reduced.The complications related with CBP treatment included aggravating anemia,thrombopenia,lower body temperature,hemorrhage,oliguria,thick sputum,pain and dysphoria.No serious lethal complication was observed.Conclusions CBP treatment is a safe and effective rescue method for ARDS in children,it can reduce pulmonary edema,improve pa (O2)/FiO2 and Cdyn,and improve mechanical ventilation parameters.During CBP therapy,the key points include monitoring vital sign and the changes of hemodynamic,keeping body position nursing care well,tranquilizing,as well as maintaining the pipe unobstructed.

Study on network compatibility of metabolisms in vivo rat for volatile oil in houttuyniae herba and 2-undecanone / 中国中药杂志

<p><b>OBJECTIVE</b>To study on the differences of metabolites in the blood of rats after injection or oral administration of volatile oil in Houttuyniae Herba and of 2-undecanone in order to validate hypothesis of network compatability and to establish the theoretical foundation of elucidating metabolic mechanism for multiple constituents in Chinese Material.</p><p><b>METHOD</b>A gas chromatography-mass spectrometry method was employed for qualitative determination of the metabolites in blood of rats by injection or oral administration of volatile oil in Houttuyniae Herba and of 2-undecanone, which were compared with the differences between binary two drugs as well as two administration.</p><p><b>RESULT</b>There were 45 kinds of ingredients in volatile oil for original materials for Houttuyniae Herba, and 53, 69 kinds of metabolites were detected in blood of rats for oral administration of volatile oil in Houttuyniae Herba, and 2-undecanone, respectively, whereas 52, 42 kinds of metabolites were detected for injection administration. Although there were various constitutes, i. e. single versus multiple ingredients, administrated by various ways, i. e. oral versus injection, the metabolites were so similar with each manners, in which alpha-pinene and beta-myrcene occurred in two constitute forms and two administrations, while there were 18, 13 kinds of ingredients for oral or injection administration of volatile oil in Houttuyniae Herba, i. e. 34%, 25% of total ingredients, respectively, and 17, 15 kinds of ingredients, i. e. 24.6%, 35.7% for that of, 2-undecanone, respectively, on average of 29.8% that suggested one of third as co-metabolites.</p><p><b>CONCLUSION</b>There are network compatibility existed in metabolism of the houttuynia volatile oil in rats, and similar metabolic pathways in accordance with original plant, among their ingredients no matter alone or multiple constitute forms and oral or injection administration.</p>

Study on differences between pharmacokinetics and chromatopharmacodynamics for Chinese materia medica formulae / 中国中药杂志

<p><b>OBJECTIVE</b>To study on the differences between chromatopharmacokinetics (pharmacokinetics with fingerprint chromatography) and chromatopharmacodynamics (pharmacodynamics with fingerprint chromatography) of Chinese materia medica formulae to answer the question whether the pharmacokinetic parameters of multiple composites can be utilized to guide the medication of multiple composites.</p><p><b>METHOD</b>On the base of established four chromatopharmacology (pharmacology with chromatographic fingerprint), the pharmacokinetics, and pharmacodynamics were analyzed comparably on their mathematical model and parameter definition.</p><p><b>RESULT</b>On the basis of quantitative pharmacology, the function expressions and total statistical parameters, such as total zero moment, total first moment, total second moment of the pharmacokinetics, and pharmacodynamics were analyzed to the common expressions and elucidated results for single and multiple components in Chinese materia medica formulae. Total quantitative pharmacokinetic, i.e., chromatopharmacokinetic parameter were decided by each component pharmacokinetic parameters, whereas the total quantitative pharmacodynamic, i.e., chromatopharmacodynamic parameter were decided by both of pharmacokinetic and pharmacodynamic parameters of each components. The pharmacokinetic parameters were corresponded to pharmacodynamic parameters with an existing stable effective coefficient when the constitutive ratio of each composite was a constant.</p><p><b>CONCLUSION</b>The effects of Chinese materia medica were all controlled by pharmacokinetic and pharmacodynamic coefficient. It is a special case that the pharmacokinetic parameter could independently guide the clinical medication for single component whereas the chromatopharmacokinetic parameters are not applied to the multiple drug combination system, and not be used to solve problems of chromatopharmacokinetic of Chinese materia medica formulae.</p>

Comparison of plaque-forming characteristics and morphogenetic changes of Newcastle disease virus isolated from chickens and geese on host cells / 病毒学报

The plaque-forming characteristics of Newcastle disease viruses of chickens and geese source were compared on various cells. The result showed that there were obvious differences of plaque formation between F48E9 and NA-1 on Vero cells, chicken embryo fibroblast cells (CEF) and goose embryo fibroblast cells (GEF). The plaque-forming ability of NA-1 was higher than F48E9 on GEF, but lower than F48E9 on CEF. On Vero cells, the plaque-forming ability of NA-1 was slightly stronger than F48E9. It demonstrated that the plaque-forming characteristics were consistent with host tropism of virus. The morphogenesis of F48E9 and NA-1 on Vero cells was observed with transmission electron microscope. There were different replication processes between F48E9 and NA-1 on cells at different stages. NA-1 had stronger adaptability to host than F48E9 according to budding processes and envelope integrity.

Solubility peak and dielectric requirement of Buyang Huanwu docoction / 中国中药杂志

<p><b>OBJECTIVE</b>To study the solubility peak and dielectric requirement of the Buyang Huanwu docoction materials, and provide theoretical and experimental foundation for selecting extraction solvent for extracting traditional Chinese drugs (TCD).</p><p><b>METHOD</b>11 types of solvents were employed as single or complex solvent systems, whose solubility parameter and dielectric constant were from 14.11 to 47.86, dielectric requirement from 1 to 80 respectively, to lixiviate Buyang Huanwu decoction (5 g per samples) in nearly saturate volume as V0 for materials at 25 degrees C. The apparent solubilities of extracts were determined and calculated out according to the section of determination of extract in the appendix of 'Chinese Pharmacopoeia'.</p><p><b>RESULT</b>The saturate solvent V0 for materials powder were 0.21, 0.31, 0.49, 0.36, 0.77, 0.93, 0.86, 0.92, 1.08, 1.00, 1.14 mL x g(-1), respectively. The apparent solubility of Buyang Huanwu docoction for each solvent system were 114.0, 101.3, 73.40, 109.4, 210.7, 295.0, 501.4, 437.0, 355.6, 423.1, 210.6 g x mL(-1), respectively, among which the max apparent solubility, illustrated as solubility peak, was carried out by methanol-water (68: 32) with 47.5 corresponding to the Buyang Huanwu docoction dielectric requirement.</p><p><b>CONCLUSION</b>The apparent solubilities of (TCD) and their formula are controlled by dielectric constant of extraction solvent, and are in accordance with stable dielectric requirement.</p>

Isolation and identification of a canine coronavirus strain from giant pandas (Ailuropoda melanoleuca)

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.

Emergency prophylactic effects of the avian influenza virus immunized serum on the infected mice / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate emergency prophylactic effects of the avian influenza virus immunized serum on experimentally infected mice.</p><p><b>METHODS</b>Serum HI antibody titers of 30 mice were detected at day 1 to 19 after being inoculated with 0.2 ml immune serum to estimate half life of immune serum. Ten mice clinical symptom was recorded to estimate the serum security after mice injected 1.5 ml immune serum. Seventy mice were randomly divided into 7 groups according to random number table and inoculated with 0.2 ml, 0.1 ml and 0.05 ml immune serum respectively via intraperitoneal injection on day 8, 4 and 1 prior to challenged with 10 LD(50) influenza virus intranasal. Mice were observed continually for 14 days to calculate the morbidity, mortality, average survival days and compare the lung index and viral titers in lung.</p><p><b>RESULTS</b>Serum HI antibody titers of mice which inoculated with 0.2 ml immune serum maintained 2(6) in 15 days after injection, but drawdown after day 17, the mice injected 1.5 ml immune serum were all alive and none onset. The survival rate of mice which injected 0.2 ml serum on the day 8, 4, 1 before challenge was 80%, 100% and 100%, and the average survival period was 13.1 days, 14.0 days and 14.0 days respectively. The survival rate of mice which injected 0.1 ml and 0.05 ml serum on day 1 before challenge was 100% and 50%, and the average survival days were 14.0 days and 11.7 days respectively. The mice lung index of experimental groups (0.0096 +/- 0.0033 - 0.0145 +/- 0.0060) was smaller than that of viral control group (0.0199 +/- 0.0025), with a statistical significance (P value 0.0022 - 0.0470, < 0.05). The viral titers in lung were significantly decreased by 2 titer as compared to the viral controls.</p><p><b>CONCLUSION</b>The avian influenza virus immunized serum might contain the emergency prophylactic effects and could be developed as an agent for possible human-avian influenza pandemic.</p>

Study on the histopathology of cats inoculated with H5N1 subtype high pathogenic avian influenza virus originated from tigers / 病毒学报

In this study, the HPAIV A/Tiger/Harbin/01/2002 (H5N1) used was originated from tigers and propagated in SPF embryonated hen eggs. TCID5, of the virus was 10(-7.36)/0. 05mL on MDCK cell. The cats were inoculated through bronchus route and then, the cats of dead and control were collected for histopathological and immunohistochemistry examination. Meanwhile, the emulsion supernatant fluid of organs and the pharyngeal swab samples of the dead cats were collected for RT-PCR, survived cats and the control cats were tested for the presence of HI antibody by standard method. The results indicated that the damage of lungs from the dead cats were most obvious, the wide range of red consolidation focus emerged on the lobus pulmonis, the fused focus of infection caused injury of lungs. Histology under the microscope revealed diffuse alveolar damage, confluence phlegmasia pathology, infiltration of lymphomonocytes, sackful of infiltration of macrophages and manipulus protein-like effusion in the alveolar. By immunohistochemistry, the positively stained virus particles were found on the epithelial cells of bronchus and alveolus, and also in the endochylema of lymphomonocytes. The specific electophoretic band of 464bp amplified by RT-PCR from samples of pharyngeal swabs, lungs, kidneys, hearts and brains was as same as the theory value. HI antibody titers of the survived cat were 1:32.

Selection of recombinant fowlpox virus coexpressing HIV-1 gag-gp120 and IL-6 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the recombinant fowlpox virus (rFPV) coexpressing HIV-1 gag-gp120 and hIL-6.</p><p><b>METHODS</b>The recombinant expressing plasmid pUTA-GE-IL6 was successfully constructed by inserting gag-gp120 gene and hIL-6 gene into the downstream of the combined promoter ATI-p7.5 and p7.5 tandem promoter respectively. After transfecting the plasmid into chicken embryonic fibroblast (CEF) cells preinfected with FPV 282E4 strain and selecting the recombinant virus under the pressure of BUdR. The recombinant virus was analyzed by nucleic acid probe hybridization and immunoblotting. In addition, the formation of virus-like particle and the expression of interested proteins in the recombinant virus-infected p815 cells were observed, and the immunogenicity of the recombinant virus was also analyzed.</p><p><b>RESULTS</b>There was colorable dot for the positive recombinant virus, immunoblotting analysis showed that the recombinant virus could expressed both gag-gp120 and IL-6. Virus-like particles (VLP) were formed in virus-infected cells, and the interested proteins could be expressed in mammalian cells infected by the recombinant virus. The immunity index from the immunized mice showed that the recombinant virus had good immunogenicity.</p><p><b>CONCLUSION</b>The recombinant fowlpox virus coexpressing gag-gp120 and IL-6 was successfully constructed, which may provide basis for the preparation of live vector genetic engineering vaccine and macromolecule particle vaccine against HIV-1.</p>