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Objective@#To explore the association between physical activity levels and depressive symptoms among college students, so as to provide evidence for promoting physical and mental health of college students.@*Methods@#Using a cohort study design,a baseline survey of 1 415 college students in Shenyang Normal University in 2017 followed for three years from 2018 to 2020. The International Physical Activity Questionnaire-Short Form (IPAQ-SF) and Selfrating Depression Scale (SDS) were used to evaluate the physical activity levels and depressive symptoms of college students. According to the level of physical activity at baseline (MET ·h/week), participants were divided into three groups [T1 (≤36.4), T2 (>36.4-89.3) and T3 (>89.3)]. Logistic regression analysis was used to examine the association between baseline physical activity levels and the incidence of depressive symptoms during the school years.@*Results@#In the 3year followup (2018-2020), 852 (60.2%) participants exhibited depressive symptoms. The numbers of individuals with depressive symptoms in the T1, T2, and T3 groups were 324, 268, and 260, respectively. Logistic regression analysis showed that college students who participated in higher levels of physical activity decreased the risk of depressive symptoms by 34.2% [T2: OR(95%CI) = 0.658 (0.500-0.866)] and 38.9% [T3:OR(95%CI)=0.611(0.465-0.804)] compared with T1 group after adjusting for demographic, lifestyle, healthrelated factors, and baseline depressive symptoms as covariates. The sexstratified analysis showed a negative correlation between physical activity levels and the incidence of depressive symptoms in female college students. Compared with T1 group, higher levels of physical activity reduced the incidence of depressive symptoms by 39.6% [T2: OR(95%CI)=0.604(0.445-0.820)] and 37.7% [T3: OR(95%CI)=0.623(0.459-0.846)], respectively (P<0.01). However, there was no significant correlation between physical activity levels and depressive symptoms in male college students (P>0.05).@*Conclusions@#There is an inverse relationship between physical activity levels and depressive symptoms. The findings suggest that schools should reduce the risk of depressive symptoms by promoting physical activity levels among college students.
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Endometritis is an inflammatory change in the structure of the endometrium due to various causes and is a common cause of infertility. Studies have confirmed that microRNAs (miRNAs) play a key regulatory role in various inflammatory diseases. However, the miRNA-mediated mechanism of endometrial inflammation induced by lipopolysaccharides (LPS) remains unclear. In this study, real-time quantitative polymerase chain reaction, Western blot analysis, immunofluorescence and Rac family small GTPase 1 (Rac1) interference were used to reveal the overexpression of miR-488 in the LPS-induced bovine uterus, and the effect of protein kinase B κ-light chain enhancement of the nuclear factor-activated B cells (AKT/NF-κB) pathway in intimal epithelial cells. The results showed that the expression of inflammatory cytokines such as interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α in the experimental group was significantly lower than that in the control group when miR-488 was overexpressed. Similar results were observed in the expression levels of p-AKT, p-IKK, and p-p65 proteins. In addition, the dual-luciferase reporter system confirmed that miRNA-488 may directly target the 3'-untranslated region of Rac1. In turn, the expression of Rac1 was inhibited. Moreover, the nuclear translocation of NF-κB was inhibited, and meanwhile, the accumulation of reactive oxygen species (ROS) in the cells was reduced. Thus, we provide basic data for the negative regulation of miR-488 in LPS-induced inflammation by inhibiting ROS production and the AKT/NF-kB pathway in intimal epithelial cells.
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Endometriosis/enzimología , Endometrio/enzimología , Células Epiteliales/enzimología , MicroARNs/metabolismo , FN-kappa B/metabolismo , Neuropéptidos/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Animales , Bovinos , Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Endometriosis/inducido químicamente , Endometriosis/genética , Endometriosis/patología , Endometrio/patología , Células Epiteliales/patología , Femenino , Regulación de la Expresión Génica , Células HEK293 , Humanos , Lipopolisacáridos , Ratones Endogámicos BALB C , MicroARNs/genética , Neuropéptidos/genética , Fosforilación , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Proteína de Unión al GTP rac1/genéticaRESUMEN
POEMS syndrome is a disease associated with plasma cell paraneoplastic syndrome. There is no unified standard treatment, and often general symptomatic treatment, drug treatment, radiotherapy, autologous hematopoietic stem cell transplantation, etc. is used. The latest treatment progress of POEMS syndrome is reviewed.
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<p><b>OBJECTIVE</b>To detect potential mutation in a family affected with congenital dyserythropoietic anemia type II (CDA II).</p><p><b>METHODS</b>Targeted sequence capture and next-generation sequencing (NGS) were used to analyze the exons and exon-intron boundaries of the SEC23B gene in a clinically suspected CDA II patient. Genotypes of the relatives were validated by Sanger sequencing. Potential impact of amino acid substitution on the structure and function of SEC23B protein was predicted with MutationTaster and PolyPhen-2. The protein structure was predicted with SWISS-MODEL software.</p><p><b>RESULTS</b>The proband was found to harbor double heterozygous mutations of the SEC23B gene, c.1727T>C (p.F576S) and c.1831C>T (p.R611W), which resulted in amino acid substitutions p.F576S and p.R611W. Both mutations were confirmed by Sanger sequencing. The sister of the proband was found to have carried c.1727T>C (p.F576S), while her father and son have carried c.1831C>T (p.R611W) mutation. In addition, the proband was detected to have carried c.211C>T (p.R71X) of the HFE gene, which resulted in substitution of arginine by a stop codon. The impact of above mutations on the structure or function of protein was predicted to be harmful. Splenectomy and iron chelation therapy have achieved effective improvement of anemia and iron overload. Computer simulation suggested that the mutations have altered the 3D structure of the SEC23B protein.</p><p><b>CONCLUSION</b>The novel compound mutations of c.1727T>C and c.1831C>T of the SEC23B gene probably underlie the CDA II in the family, and there is a strong correlation between the genotype and phenotype.</p>
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Adulto , Femenino , Humanos , Anemia Diseritropoyética Congénita , Genética , Simulación por Computador , Familia , Genotipo , Secuenciación de Nucleótidos de Alto Rendimiento , Mutación , Fenotipo , Proteínas de Transporte Vesicular , GenéticaRESUMEN
POEMS syndrome is a kind of plasma cell disorder with multi-system involved,whose character includes polyneuropathy,organomegaly,endocrinopathy,monoclonal plasma cell disorder,as well as skin changes.Its pathogenesis is not clear,with low incidence,high misdiagnosis rate and high disability rate.There are still a variety of dispute on its diagnosis.Here this article reviewed the diagnostic progress of POEMS syndrome recent years.
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[Objective] The objective of this study was to evaluate the diagnostic accuracy of the 2013 edition of Breast Imaging Reporting and Data System (BI-RADS) ultrasound lexicon in diagnosing breast categories 3-5 lesions.[Methods] Using our breast ultrasound database from June 2014 to June 2016,we identified 4428 BI-RADS category 3 to 5 lesions with a known pathological diagnosis in 4 428 adult women.The positive predictive value (PPV) of each BI-RADS category was calculated based on the pathological diagnoses and compared with the reference range provided by the American College of Radiology (ACR).[Results] 4 428 lesions from 4428 patients were included in this study.The PPV of each BI-RADS category waswithin the reference range provided by the ACR in 2013.1198 (27.1%) pathological malignant/borderline results were found in the 4 428 lesions,the other 3 230 (72.9%)lesions were diagnosed with benign results.Among the malignant/borderline lesions,the rate of lymph node metastasis gradually increased as the BI-RADS categories were upgraded.Malignant lesions with a diagnosis ofinvasive ductal carcinoma or invasive lobular carcinoma showed an increasing distribution trend from category 3 to 5.[Conclusion] The 2013 editionof BI-RADS ultrasound lexiconhas good diagnostic accuracy and efficiencyin clinical practice.
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AIM: To study the effects of disulfiram/copper complex (DSF/Cu) on ultrastructures and mechan-ical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy (AFM) based on the nanoscale resolution and piconewton force measurement level.METHODS: The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10A cells induced by DSF/Cu were compared by flow cytometry.The cell surface morphology, ultra-structure, height, width and roughness were detected by AFM.The effects of DSF/Cu on the hardness (Young’s modu-lus) of the 2 kinds of cells were determined by AFM with indentation technique.RESULTS: DSF/Cu significantly in-duced apoptosis of the MCF-7 cells in a dose-dependent manner, whereas had little effect on the MCF-10A cells.The cell cycle analysis showed that DSF/Cu induced G2 /M arrest in the MCF-7 cells, but led to G0 /G1 arrest in the MCF-10A cells.The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology, and the filopo-dia were retracted obviously, even some became into lamellipodia, or disappeared completely after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.The quantitative analysis indicated that the MCF-7 cells showed smaller width and larger height, and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.However, little effect in the MCF-10A cells was observed.The biomechanics test at a single cell level demonstrated that the Young’s modulus of the MCF-7 cells and MCF-10A cells were both increased, yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10A cells after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.CONCLUSION: DSF/Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.
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Aim To study the mechanism of DSF-Cu induced apoptosis of human nasopharyngeal carcinoma CNE-2 Z cells by affecting the function of mitochondria and cytoskeleton. Methods The cell cycle,the rate of apotosis,the levels of intracellular ROS and MMP in CNE-2 Z cells were tested by flow cytometry after trea-ted with different concentration of DSF-Cu. The chan-ges of the cell surface morphology, ultrastructure, cell height, width and roughness were detected by AFM. The distribution and reorganization of cytoskeleton F-actin were observed by Laser scanning confocal micro-scope. Results Cells were incubated with different concentration of DSF-Cu ( 0 ~200 nmol · L-1 ) for 24 h, the apoptotic ratio increased significantly and the treatment of DSF-Cu resulted in a concentration-de-pendent accumulation of CNE-2Z cells in G2/M phase. Furthermore,the treatment of DSF-Cu was able to in-crease the production of intracellular ROS and decrease the MMP in CNE-2Z cells. In addition,AFM imaging showed that compared to the control group,with the in-crease of DSF-Cu concentration,the CNE-2Z cells be-came smaller, cytoplasm condensed, the height in-creased,and the surface roughness reduced. Moreover, the filopodia became shorter, shrinked and even com-pletely destroyed after treated with different concentra-tion of DSF-Cu. At last,the LSCM image showed that the fluorescence intensity of F-actin networks was de-creased, then the structure was rearranged and de-stroyed obviously by treated with DSF-Cu. Conclusion DSF-Cu can induce apoptosis and arrest cell cycle at G2/M phase in CNE-2Z cell through a mitochondria-dependent pathway. Above findings highlight the appli-cations of AFM at the single cell level for the investiga-tion of antineoplastic drug in nasopharyngeal carcinoma therapy.
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<p><b>BACKGROUND</b>Autophagy has been found to be involved in animal and cell models of atherosclerosis, but to date, it lacks general observation in human atherosclerotic plaques. Here, we investigated autophagy in smooth muscle cells (SMCs), endothelial cells (ECs), and macrophages in human atherosclerotic plaques via transmission electron microscopy (TEM), western blotting, and immunohistochemistry analysis.</p><p><b>METHODS</b>The histopathologic morphology of these plaques was observed via hematoxylin and eosin staining. The ultrastructural morphology of the SMCs, ECs, and macrophages in these plaques was observed via TEM. The localization of microtubule-associated protein 1 light chain 3 (MAP1-LC3), a relatively special maker of autophagy, in plaques was observed by double fluorescent immunochemistry and western blotting.</p><p><b>RESULTS</b>All of these human atherosclerotic plaques were considered advanced and unstable in histologically observation. By double fluorescent immunochemistry, the expression of LC3-II increased in the SMCs of the fibrous cap, the macrophages, and the microvascular ECs of the plaque shoulders. The protein level of LC3-II by western blotting significantly increased in plaques compared with normal controls. In addition, TEM observation of plaques revealed certain features of autophagy in SMCs, ECs, and macrophages including the formation of myelin figures, vacuolization, and the accumulation of inclusions in the cytosol. These results indicate that autophagy is activated in SMCs, ECs, and macrophages in human advanced atherosclerotic plaques.</p><p><b>CONCLUSIONS</b>Our study is to demonstrate the existence of autophagy in human atherosclerotic plaques by different methods, which may contribute to the development of pharmacological approaches to stabilize vulnerable and rupture-prone lesions.</p>
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Humanos , Aterosclerosis , Metabolismo , Autofagia , Fisiología , Células Endoteliales , Patología , Técnicas In Vitro , Microscopía Electrónica de Transmisión , Proteínas Asociadas a Microtúbulos , Metabolismo , Miocitos del Músculo Liso , Patología , Placa Aterosclerótica , MetabolismoRESUMEN
Objective To detect the change of serum cystatine C ( cys C ) level in Parkinson ’ s disease ( PD ) patients with obstructive sleep apnea-hypopnea syndrome ( OSAHS ) and explore its influencing factors .Methods Fifty-six PD patients with polysomnography examination from July 2011 to December 2013 in the Department of Neurology , the Second Affiliated Hospital of Soochow University were collected.Eighteen healthy controls who took the polysomnography examination during the same period were included.According to the apnea-hypopnea index ( AHI) , PD patients were further divided into two groups:PD with OSAHS group ( n=26 ) , and PD group ( n=30 ).The general conditions , movement function , biochemistry parameters , and sleep parameters were assessed.Statistical analysis was performed using SPSS ver 17.0 software.Results The mean serum levels of cys C in PD with OSAHS group , PD group and control group were (1.05 ±0.17) mg/L, (0.96 ±0.12) mg/L and (0.84 ±0.20) mg/L, respectively.Statistical analysis showed that there was significant difference in the three groups (F=9.184,P0.05).Multivariate analysis showed that creatinine levels (B=0.007,P=0.005), AHI (B=0.004,P=0.013) , H-Y stage (B=0.102,P=0.026) may be influencing factors of cys C levels in PD patients (P<0.05).Conclusions Cys C level is elevated in PD patients, especially in PD patients with OSAHS.The degree of hypoxia and severity of PD is related to the level of cys C in PD patients with OSAHS .
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Objective To explore the correlation between the syndromes of patients with gastric carcinoma and level of serum sE-cad. Methods Totally 190 cases of different syndromes of patients with gastric carcinoma were collected, and serum level of sE-cad was measured by ELISA. The correlation between the syndromes of gastric cancer and level of serum sE-cad was analyzed by SPSS16.0 software. Results The level of serum sE-cad of interior retention of blood stasis syndrome was significant different with that of deficiency syndrome of spleen and stomach, yin impairment syndrome due to stomach heat, deficiency syndrome of both qi and yin (P<0.05). Besides, this result was the same as the results of gender stratification analysis in men. The difference in serum level of sE-cad in the type of deficiency syndrome of spleen and stomach was significant different with that of deficiency syndrome of both qi and yin in women (P<0.05). Conclusion Level of serum sE-cad was correlated with syndrome type of patients with gastric carcinoma.
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Aim To study the protective effects of Paeoniflorin (PF) on dopaminergic neurons in brain slice of substantia nigra treated with MPP~+ and to investigate the transcription of alpha-synuclei (α-syn) mRNA.Methods The organatypic brain slice culture of substantia nigra prepared from neonatal SD rats was placed on Millicell-CM porous membranes and cultured to day-10.Then the cultrues of slice were treated with different concentrations (0.1,0.5,1.0 mmol·L~(-1)) of MPP~+ for 24 h.Some of the cultrues treated with 0.5 mmol·L~(-1) MPP~+ also received PF (1 or 10 μmol·L~(-1)).Slices cultured in normal medium were used as vehicle control.The tyrosine hydroxylase (TH) immunohistochemical staining with the cell counting was used to determine the dopaminergic neruons.The transcription of α-syn mRNA was examined by real-time quantitative RT-PCR.Results MPP~+(0.1,0.5,1.0 mmol·L~(-1)) exposure markedly decreased the number of TH~+ cells in a dose-dependent manner (P<0.05 or P<0.01) and sharply induced the transcription of α-syn mRNA (P<0.01) in slices treated with 0.5 mmol·L~(-1) MPP~+.The addition of PF (10 μmol·L~(-1)) to MPP~+-treated slices significantly increased dopaminergic neurons survival (P<0.01) and downregulated the transcription of α-syn mRNA significantly (P<0.05).Conclusion PF can effectively inhibit the injury of dopaminergic neurons induced by MPP~+ on brain slice of substantia nigra and downregulate the transcription of alpha-synuclein.
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Objective To explore the specific role of autophagy and ubiquitin-proteasome pathway in apoptosis, specific protease inhibitor and (or) macroautophagy inhibitors.Methods The stimulators were selected to work on the pheochromocytoma (PC12) cell lines transfected with human mutant α-synuclein (A53T).Cell activity and apeptosis rate were detected by MTT law and flow cytometry.NO energy, heat shock protein 70 (Hsp70) and Caspase-3 expression were determined in cell culture.Results A53T cell survival rate significantly decreased 24 hours after handling with the protease inhibitor (100 nmol/L) and (or) autophagy inhibitors 3-MA (10 mmol/L, A =0.23±0.01,0.19±0.01 and 0.17±0.01 respectively; P <0.05) compared with the control group (A =0.32±0.06).Cell survival rate was significantly higher than the other drug group after 24 hours handling with autophagy stimulators (A =0.44±0.08).Compared with the control group or autophagy stimulator of rapamycin (0.2 μg/ml) group (1.55%±1.15%), A53T cells apeptosis percentage rate was significantly higher after treated with proteasome inhibitor and macroautophagy inhibitors 24 hours (4.74%±0.91%, 4.59%±1.18% and 5.40%±1.75%respectively, P <0.05); and a slight decrease with stimulators.Protein Hsp70 and NO were significantly higher in proteasome inhibitor groups than the control group.But in antophagy inhibitor and stimulator group, NO and Hsp70 protein was similar to the control group.Conclusion The inhibition of macroautophagy and proteasome can promote apoptosis.Inhibiting or stimulating autophagy has less impact on Hsp70 and NO than proteasome pathway.
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Objective To explore the protective effect and mechanism of gardenoside on the damage of dopaminergic neurons induced by lipopolysaccharide (LPS). Methods Both neuron-enriched cultures and neuron-astrocyte cultures were pretreated with vehicle or gardenesides ( 10, 20 and 40 mg/L) for 30 min at 37℃. The culture media were subsequently renewed in order to remove gardenesides. LPS was then added into all culture media at a final concentration of 10 mg/L Twenty-four hours later, the culture media was collected to measure TNF-α, NO, IL-6, GDNF and MMP-9; the cells were collected to count the number of cells labeled with an antibody against tyrosine hydroxylase (TH) and to assess the expression of TH mRNA using reverse transcription-polymerase chain reaction. Results Gardeneside didn't promote the survival of dopaminergic neurons in neuron-enriched culture, but significantly increased the survival of dopaminergic neurons in neuron-astrocyte culture, compared with the vehicle group, the survival of dopaminergic neurons increased from 203.0%±17.4% to 256.7%±15.2% ( F = 17.22, P = 0.001 ) in 40 mg/L gardenaside group. The amount of TNF-α, NO and GDNF released from the neuron-astrocyte cultures after 24 h of addition of LPS was not changed significantly, while the expression of IL-6 and MMP-9 was increased significantly. In this study, the gardenoside concentration-dependently attenuated the LPSinduced increase of the expression of IL-6 and MMP-9, compared with the vehicle group, the expression of IL-6 and MMP-9 decreased to 67.2%±6.4% (F= 12.89,P =0.001 ), 77.3%±9.8% (F =8.27,P = 0.001 ) respectively in 40 mg/L gardenoside group. Conclusions Astrocytes play a neuroprotective role on dopaminergic neurons, which is decreased by LPS via inducing the secretion of pro-inflammatory factors. Gardeneside may protect dopaminergic neurons from LPS-induced injury in an astrocyte-dependent manner and it inhibits the production of proinflammatory factors instead of promoting the secretion of GDNF. From the point of view that a very low toxicity of gardenesides has been well documented, this report may reveal a new way of developing therapeutic interventions for inflammation-related diseases such as Parkinson's disease.
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Objective To investigate the role of astrocyte in the lipopolysaccharide-induced damage of dopaminergic neurons. Methods After lipop01ysaccharide was applied to the third generation of rat astrocytes for 24 hours,supernatants of astrocytes culture were collected.The primary middle-brain dopaminergic neuron-enriched culture systems were obtained by neurobasal and ara-c,eoeulture system of both astrocytes and neurons was established by transwell inserts.The lipopolysaccharide was administered into neuron-enriched systems and coculture systems and the change of dopaminergic neurons was detected.At the same time,the supernatants of astrocytes were administered into the neuron-enriched systems,and the survival of dopaminergic neurons and the expression of tyrosine hydroxylase mRNA were observed. Results Lipopolysaccharide had a negative effect on the survival of dopaminergic neurons in a concentration-dependent manner.Both astrocytes and supernatants promoted the survival of dopaminergie neurons,and the former was better than the latter. In the preoccupation of existence of astrocytes,low-concentration lipopolysaccharide promoted the survival of dopaminergic neurons,while high-concentration,decreased.The change of the expression of tyrosine hydroxylase mRNA was similar to the survival of dopaminergic neurons.Conclusions Astrocytes play a protective role in the damage of dopaminergic neurons induced by lipopolysaccharide,and suitable activation of astrocytes would increase the protective effect while excessive activation of astroeytes would attenuate the effect.
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Objective To observe the effect of mutant α-synuclein(A30P)in autophagic programmed cell death by transfected PC12 cells and explore its probable role and pathway in PD.Methods The definite PC12 cells which were transfected mutant α-synuclein(A30P)were constructed at first and MPP+,Rapamycin and Wortmanin were administrated to transfected PC12 cells with mutant α-synuclein. Not only the proliferative activity of cells was detected with MTT method but also the ultrastructttre changes of cells and expression of α-synuclein in different circumstance were observed by transmission electron microscopy(TEM),Western Blot and the level of SOD.Results (1)The expression of α-synuclein in groups A30P+Wortmannin and A30P+MPP+was higher than that in group A30P(P<0.01), particularly.there was more significant expression of α-synuclein in group A30P+Wortmannin.The expression of α-synuclein in group A30P+Rapamycin was weaker than that in group A30P(P<0.01); (2)The results showed that the SOD level(group A30P+MPP+:3 h:97.49±13.8;12 h:102.7±12.7; 24 h:101.5±11.8;48 h:104.3±12.4)was significantly decreased at various time points after MPP+ treatment compared that of group A30P(t=3.7721,P=0.0017).SOD level gradually increased in A30P +Rapamycin 12 h and showed significant difference at 24 h(121.2±13.0),48 h(124.3±14.1)and 72 h(127.7±13.7)after drug treatment compared with that in group A30P+Wortmannin(t:2.9746, P=0.0083);(3)Mutant α-synuclein(A30P)leading to PC12 cells death by means of autophagy involved α-synuclein accumulation,membrane lipid oxidation,and loss of plasma membrane integrity.Mutant α- synuclein(A30P)mediated the toxicity of MPP+.Rapamycin,an inducer of autophagy,reduced the aggregation of α-synuclein in transfected cells.Meanwhile,Wortmanin,an inhibitor of autophagy,promoted the aggregation of α-synuclein in transfected cells and induced cells to die.Conclusions The abnormal aggregation of α-synuclein induces autophagic programmed cell death in PC12 cells and mutant α-synuclein (A30P)mediates the toxicity of MPP+.Meanwhile,Rapamycin may reduce the aggregation of α-synuclein in transfeeted cells by activation of autophagic pathway.
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The paper discusses the connotation of the medical ethics and style,and analyzes the problems existed in present medical ethiCS construction.Based on systemic thinking on the medical ethics construction,it proposes that from the height of hospital culture construction and under the principle of combining the belief cultivation with practice of medical ethics,the management mechanism involving education,supervision,examination and evaluation,rewards and punishment should be established.
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Cobra venom secretory phospholipase A_2 (sPLA_2) is an important component of cobra venom which has a variety of biological activities. Recent studies are mainly focusing on each pharmacological active component of venom, SPLA_2 is one of them. This review summarized the structure, purification and biological activities of cobra venom sPLA_2 with emphasizing its diverse pharmacological effects and toxicity. In addition, some mechanisms of actions of sPLA_2 and possible applications of sPLA_2 were also discussed.
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Objective To obtain an antigenic gene of adult Trichinella spiralis. Methods cDNA library of the adult Trichinella spiralis was screened using the sera of immunized and infected rabbits. The gene sequence was analyzed by DNAstar software and GenBank database. Results Nine positive clones were identified by immunoscreening. The clone Ts87 was sequenced and a cDNA with 1 172 bp full length was obtained using 5′ RACE technique, encoding 347 amino acids. Some possible antigen epitopes were predicted. Conclusion A novel antigenic gene of Trichinella spiralis was obtained.
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Objective To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein. Methods Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET 28c(+) expression vector and expressed in E.coli . Mice were immunized with the fusion protein incorporated into Freund’s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm. Results The fragment of Ts88 gene was expressed successfully in E.coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm. Conclusion The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity.