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BackgroundPreclinical and clinical evidence suggests that hyperbaric oxygen therapy (HBOT) may benefit newborns. The effectiveness of HBOT for neonatal hypoxic-ischemic encephalopathy (HIE) remains controversial. We conducted a meta-analysis to evaluate the efficacy and prognosis of HBOT in neonates with HIE.MethodsA systematic search of eight databases was performed for available articles published between January 1, 2015, and September 30, 2020, to identify randomized controlled clinical trials (RCTs) on HBOT for neonatal HIE. Methodological quality assessment was performed by applying the simple procedure detailed by the Cochrane collaboration. Afterward, quality assessment and data analysis were performed using Revman 5.3 software. STATA 15 software was used to detect publication bias as well as for sensitivity analysis.ResultsA total of 46 clinical RCTs were selected for the study and included 4,199 patients with neonatal HIE. The results indicated that HBOT significantly improved the total efficiency (TEF) of treatment for neonatal HIE patients [odds ratio (OR) = 4.61, 95% confidence interval (CI) (3.70, 5.75), P < 0.00001] and reduced the risk of sequelae (OR = 0.23, 95% CI (0.16, 0.33), P < 0.00001) and the neonatal behavioral neurological assessment (NBNA) scores [mean difference (MD) = 4.51, 95%CI (3.83,5.19, P < 0.00001)].ConclusionIn light of the effectiveness of HBOT neonatal HIE, this meta-analysis suggested that HBOT can be a potential therapy for the treatment of neonatal HIE. Due to the heterogeneity of studies protocol and patient selection being only from China, more research is needed before this therapy can be widely implemented in the clinic.Protocol RegistrationPROSPERO (ID: CRD42020210639). Available online at: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020210639.

By introduction of trilacunary Keggin-type polyoxometalate to the hydrothermal reaction system of Cr3+, Ln3+ and phthalic acid, a series of novel {Cr4Ln4} heterometallic clusters with the formula Cs2[Cr4Ln4(μ4-O)4(μ3-O)4(C8H4O4)4(H2O)12](H3SiW12O40)Cl·23H2O (1-Ln, Ln = Ce, Pr, Nd) and [Cr4Ln4(μ4-O)4(μ3-O)4(C8H4O4)4(H2O)10](H6SiW12O40)Cl2·18H2O (2-Ln, Ln = Sm, Eu, Gd, Tb, Dy, Ho, Er) have been obtained. Single-crystal structural analyses show that 1-Ln and 2-Ln constitute the first cases of Cr–Ln heterometallic clusters stabilized by inorganic polyoxometalate anions and organic ligands. Optical spectra studies demonstrate that 1-Ln and 2-Ln are narrow-gap semiconductors with band gaps of about 1.5 eV. Magnetic investigation shows that compound 2-Dy is a potential single molecule magnet.

Four seven-nuclearity 3d-4f heterometallic cluster {Fe6LnO28} substituted polyoxometalates (HPz)11K4Fe6Ln(μ3-O)2(B-α-GeW9O34)2(GeW6O26)· xH2O (1-Ln, Pz = piperazine, Ln = Tb, Dy, Ho, Er for x = 27, 25, 25, 24, respectively) have been hydrothermally prepared and structurally characterized by single-crystal X-ray diffraction, powder X-ray diffraction, infrared spectrometry, thermogravimetric analyses, elemental analyses, and electrospray ionization mass spectrometry. Single-crystal X-ray diffraction analyses revealed that 1-Ln contain an unprecedented banana-shaped polyanion constructed from an iron-lanthanide heterometallic {Fe6LnO28} cluster, two trilacunary {B-α-GeW9O34} units, and one hexalacunary {GeW6O26} fragment. The magnetic susceptibility surveying proved the presence of antiferromagnetic coupling in 1-Ln.

A series of heterometallic cluster organic frameworks based on cuprous-halide Cu4I4 clusters and lanthanide clusters, [Ln2(H2O)8]2(Cu4I4)(pdc)4][NO3]4·solvent (1-Ln, Ln = Y, Tb, Eu, H2pdc = 3,5-pyridinedicarboxylic acid), have been successfully synthesized under solvothermal conditions. Single-crystal X-ray diffraction analysis reveals that 1-Ln show fascinating three-dimensional porous framework structures constructed from two dissimilar types of nanosized coordination cages. Gas sorption measurements indicate that 1-Ln show moderate gas uptake capacities. The UV/vis spectroscopy measurements reveal that 1-Ln are a class of potential materials for the removal of Cr2O72− from aqueous solution. Additionally, the luminescence properties of 1-Tb and 1-Eu have also been investigated.

A combination of microbial strain improvement and statistical optimization is investigated to maximize echinocandin B (ECB) production from Aspergillus nidulans ZJB-0817. A classical sequential mutagenesis was studied first by using physical (ultraviolet irradiation at 254 nm) and chemical mutagens (lithium chloride and sodium nitrite). Mutant strain ULN-59 exhibited 2.1-fold increase in ECB production to 1583.1 ± 40.9 mg/L when compared with the parent strain (750.8 ± 32.0 mg/L). This is the first report where mutagenesis is applied in Aspergillus to improve ECB production. Further, fractional factorial design and central composite design were adopted to optimize the culture medium for increasing ECB production by the mutant ULN-59. Results indicated that four culture media including peptone, K2HPO4, mannitol and L-ornithine had significant effects on ECB production. The optimized medium provided another 1.4-fold increase in final ECB concentration to 2285.6 ± 35.6 mg/L compared to the original medium. The results of this study indicated the combined application of a classical mutation and medium optimization can improve effectively ECB production from A. nidulans and could be a promising tool to improve other secondary metabolites production by fungal strains.

Microchiroptera (microbats) is a suborder of bats thought to have degenerated vision. However, many recent studies have shown that they have visual ability. In this study, we labeled neuronal nitric oxide synthase (nNOS)-the synthesizing enzyme of the gaseous non-synaptic neurotransmitter nitric oxide-and co-localized it with calbindin D28K (CB), calretinin (CR), and parvalbumin (PV) in the visual cortex of the greater horseshoe bat (Rhinolophus ferrumequinum, a species of microbats). nNOS-immunoreactive (IR) neurons were found in all layers of the visual cortex. Intensely labeled neurons were most common in layer IV, and weakly labeled neurons were most common in layer VI. Majority of the nNOS-IR neurons were round- or oval-type neurons; no pyramidal-type neurons were found. None of these neurons co-localized with CB, CR, or PV. However, the synthesis of nitric oxide in the bat visual cortex by nNOS does not depend on CB, CR, or PV.

By adopting a trilacunary [A-α-GeW9O34]10− as a precursor under hydrothermal conditions, a series of novel 4s–4f cluster substituted polyoxometalates (POMs) containing two types of in situ generated lacunary POM fragments have been made. These compounds were characterized by single-crystal X-ray diffractions, powder X-ray diffractions, infrared spectroscopy, thermogravimetric analyses, respectively. Structural analyses indicate that these compounds constructed from a seven-nuclearity K-O-Ln (Ln = Dy, Ho, Er) heterometallic cluster stabilized by mixed dilacunary Keggin [α-GeW10O37]10−/multilacunary open Wells-Dawson [Ge2W18O66]16− fragments. Furthermore, the solid luminescent properties of the compounds at room temperature were studied.

Purpose: To search for the change of fibrinolytic activity of different forms of nattokinase when they were treated in human digestive juice, artifical gastric and intestinal juice differently, and to lay the foundation of a proper pharmaceutical dosage form of nattokinase. To provide direct evidences that Nattokinase products can be administered orally and to screen a suitable dosage form for oral administration of nattokinase.Methods: The non-crushed sample was prepared through the solid fermentation of soybean meal at 37°C for 36 h, and evaporation at 70°C for 8 h, The fine and ultra-fine powder were gotten from the non-crushed sample by the micronizer and ultra micronizer. Nattokinase extract was the supernatant liquid of 0.9% saline of non-crush samples. The nattokinase’s fibrinolytic activity of non-crushed sample, fine powder, ultra-fine powder and extract treated in artifical and human digestive were measured by the Fibrin plate method. Results: Three kind of solid form samples still remain about 80% fibrinolytic activity in artificial gastric juice for 4 h, and the fibrinolytic activity of nattokinase extract is 48.7% at the end of treatment. Treated in artificial gastric juice for 4 hours, then handled in artificial intestinal fluid for 4 h, the final fibrinolytic activity of fine and ultra fine powder is 47.0% and 51.0%. The fibrinolytic activity of extract is 79.8% at the end of treatment in human digestive juice for 4 h. At the beginning of treatment in human digestive juice the enzyme activity of fine and ultra fine powder is 130.8U/mL and 132.1U/mL, however, after treatment for 4 h the enzyme activity is 158.7U/mL and159.6U/mL, which is near to the total enzyme activity of sample. Conclusion: Nattokinase products can be administered orally, and solid preparations of nattokinase are more suitable for oral route. The optimum dosage form for oral administration of nattokinase is enteric-coated capsules containing ultra fine powder.

Purpose To prepare Grifola frondosa fermented beverage and study on its production process, ingredients and characteristics. Methods The following process was used to prepare Grifola frondosa fermented beverage: Grifola frondosa original strains →Strain activation →Inclined plane strain →Liquid spawn →Submerged fermentation →Fermentation broth with mycelium →Colloid mill grinding →High pressure homogenization →Homogenate →Add preservatives or not →Package → Sterilizing or not →Finished product. Grifola frondosa mycelium biomass was determined by Weight-Drying method. Total sugar content was measured by Phenol-sulfuric acid method. Reducing sugar content was measured by 3, 5-dinitrosalicylic acid (DNS) method. Protein content was measured by Coomassie brilliant blue method. The quality of Grifola frondosa fermented beverage was evaluated every 10 days from six areas: color, precipitation, transparency, aroma, taste and the bacteria growth situation. Results Mycelia biomass of Grifola frondosa fermentation product is 378.3 ± 7.21mg/100ml. The total sugar content is 1.8995 mg/ml, the reducing sugar content is 0.2539 mg/ml, and grifolan content is 1.6456 mg/ml, protein content 7.971μg/ml. The total number of colonies is 67 CFU/ml and no E. coli in Grifola frondosa fermented beverage, this comply with the requirements of the national food safety standard. The samples of Grifola frondosa fermented beverage without sterilized and preservatives placed at room temperature for 6 months are normal, and no bacteria growth. Conclusion The production process of Grifola frondosa fermented beverage is feasible. In the case without sterilization and preservatives, Grifola frondosa fermented beverage can be placed at room temperature for 6 months without deterioration and still maintain its characteristics of nutrient-rich, pure taste, clean and hygienic, with natural mushroom fragrant. This can provide a reference for the development of Grifola frondosa submerged fermentation products.

Purpose To develop a novel complex antimicrobial agent and determine the optimal components of the composite antimicrobial agents and its antimicrobial activity in vitro. Methods According to antimicrobial mechanisms，antibacterial spectrums，physical and chemical properties and applicabilities of existing antimicrobial agents in clinical use, select out cefoperazone sodium, sulbactam sodium and cephradine as the basic components to make a novel complex antimicrobial agent. Utilize yeast, staphylococcus aureus and E. coli bacteria as test bacteria. Do the three factors four-level orthogonal experiments by the maximum amount, the middle amount, low amount and Minimum amount of the three-component agent to research the optimum ratio of the drug. Measure the titer of the compound antimicrobial agent by the way of tube-plate method (2 doses). With known contents of Penicillin Sodium for Injection as control, and determine its minimum inhibitory concentration against staphylococcus aureus, E. coli and yeast by using the agar doubling dilution method. The experimental results were analysized by statistical analysis software SPSS16.0. Results The results of the three factors four-level orthogonal experiments indicate the optimum ratios of Cefoperazone Sodium, Sulbactam Sodium and Cephradine against E. coli, yeast and staphylococcus aureus were 2:2:3, 1:2:2 and 2:6:5, their titers were 1353.9U/mg, 982.7U/mg and 1015.5U/mg. With the highest titer proportion 2:2:3 as the composition of the antimicrobial compound. This compound antimicrobial agent had a good antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria and Fungi, its minimal inhibitory concentration (MIC) against staphylococcus aureus, E. coli and yeast were 2.000μg/ml, 0.500μg/ml and 16.000μg/ml. Conclusion This research acquires a composite of antibiotics. This antimicrobial compound has a broader spectrum and higher antimicrobial activity in vitro comparing with traditional common single antibiotics, and it especially has a good antimicrobial activity against fungi. The results set a scientific foundation for enriching clinical medicines.

Objective In order to increase the Paeonol dissolution and content, cortex moutan were smashed into nanoparticles, and the dissolution and content were compared by microscopy before and after super-micro-particle pulverization. Methods Super-micro-particle pulverization and general grinding were used to broke Cortex moutan into particles. The microscopic morphous characteristics of the before- and after- ultra-disintegration particles were compared by microscopy. Methods of HPLC was used to determine the content and dissolution of Paeonol with different grinding conditions. Methods of precipitation and funnel way were used to examine the stability and fluidity of cortex moutan nano-particles. Results Cortex moutan powder after super-micro-particle pulverization appears sphere or like-sphere, and its average size is 200nm~300nm. After the superfine grinding Paeonol dissolution increases 76.19% in comparison with without nano pulverization. The nanoparticle rest angle is θ=33°.The precipitation ratio of Cortex moutan powder with general grinding is 0.28 at 24h, and the precipitation ratio of its nano-power has been to 0.98 at 60min. Conclusion Paeonol dissolution, stability and fluidity of Cortex moutan nanoparticles were improved greatly and this nanoparticles is beneficial to industrial production for traditional Chinese medicine.

To prepare out a novel preparation of liuweidihuang which effectively preserves Paeonol, Ursolic acid and Polysaccharides, and the preparation process should be scientific, lower-price and simple. The microscopic characteristics and its physicochemical properties of super-micro-particle of liuweidihuang from above process were also discussed. Methods According to the therapeutic and physicochemical properties of materials in liuweidihuang, the processed technology is as follow: Ripe rehmannia glutinosa,Tuckahoe and Alisma digitalis were boiled for 4.5h., the root bark of the peony tree(Paeonia suffruticosa), dogberry and yam were smashed to nanoparticles. Then select the reasonable method to drying the mixture. The microscopic shape characteristics of the before- and after- broken particles was compared. Paeonol content and dissolution were determined by HPLC. The stability and fluidity of liuweidihuang nanoparticles were examined by precipitation and funnel way. Results The approach of prepared liuweidihuang nano-microcapsules is scientific, lower-price and simple. The average size of processed super-micro-particle of liuweidihuang is 400±46nm. The hardness of the table is 9.7kg/cm2. The thickness of the table is 0.6cm. The friability of the tablet is ＜1%. Each tested item complied with standards of troches of Chinese medicine, and it is beneficial to industrial production for Chinese medicine.. Conclusion The nano-microcapsule is dispersive, desiccative, mobilizable, stable and three times concentrated than traditional pills of Liuweidihuang. It also can be used to produce different forms of drugs in the market.

Purpose To search for an isolation method of grifolan synthase, and research grifolan synthase partly enzymatic properties. Methods Taking Grifola frondosa as material, the mycelium was broken by ultrasonic, and then centrifugated, the supernatant was collected, GS was precipitated by different concentrations of ethanol, ammonium sulfate and acetone, the suitable isolation method of GS was researched out by taking enzyme activity and protein content as parameters. Reducing sugar, protein content and total sugar content were determined respectively by using 3,5-dinitrosalicylic acid (DNS) method, Coomassie brilliant blue method and phenol-sulfuric acid method. Taking glucose as substrate, GS activity was reflected by the consumption of glucose. Consumption of glucose was measured by DNS method. The optimal temperature and pH of GS enzyme reaction was determined by carrying out the enzyme assay at different temperatures and pH levels.The acid-base stability of GS was determined by subjecting GS to different pH levels for 60 minutes, and the heat stability of GS was determined by subjecting GS to different temperatures for 30 minutes. The direction of GS enzymatic reaction was determined by measuring the consumption of β-glucan in 1 minute. The possibility of GS existing extracellular was judged by determining GS activity in extracellular fermentation liquor. Results The proper isolation method of GS is that the mycelium was collected from fermentation liquor and broken by ultrasonic for 1min, then centrifugated at 5000rpm for 10 minutes and a supernatant was collected. Ammonium sulfate was added to the concentration of 60%, and then centrifugated at 12000rpm for 10 minutes at 4°C, the pellets was collected as GS crude enzyme. Using this method, 93.86mg of crude enzyme which enzyme activity was 5700U/mg was obtained from 100g mycelium with moisture content of 87.28%, extraction rate of crude enzyme was 0.7379%. The optimum pH of GS enzyme reaction was pH=5.0 and the optimum temperature was 15°C, GS was most stable at pH=5.0 and in the range of 30 °C to 50 °C. Just 0.6996μg β-glucan was hydrolyzed in 1 minute by GS which can actually consume 5700μg glucose per minute in the synthetic reaction of β-glucan, considering the error in actual measurement, it can be considered that GS is a one-way enzyme that it can only catalyze the synthesis of β-glucan. GS activity in extracellular Grifola frondosa fermentation liquor was -0.1875U/ml, indicating that GS is one kind of intracellular enzyme and without GS activity in fermentation liquor or extracellular. Conclusion An isolation method of grifolan synthase form the mycelium of Grifola frondosa was researched out, and grifolan synthase partly enzymatic properties were studied in this paper. It can lay the foundation for the further study on the structure and function of GS and grifolan production.

：PurposeThe physicochemical properties of four Cross-linked sodium hyaluronate gels(CHA) with different cross-linking agents were conpared.In order to research out the different stability and Enzyme-resistant properties of these CHA.Methods The CHA hydrogels were prepared with different cross-linking agents, such as PEG20000, PDE, BDDE and ADH. The optimal reaction conditions were determined by single factor experiment. Dynamic viscosity was tested by Stabinger method. Intrinsic viscosity was tested by Uzziah's viscosity method. The in vitro Enzyme-resistant properties of CHA-gels were analysed by carbazole and spectrophotometry. Results The concentrations of NaOH/HCl, concentrations of HA and the ratio of cross-linking agent to HA were significant conditions which influenced the physicochemical properties of CHA gels. PDE-CHA and PEG20000 gels have best Dynamic viscosity, PDE-CHA gel has best Intrinsic viscosity, ADH-CHA and BDDE-CHA gels have better Enzyme-resistant properties than PEG20000-CHA and PDE-CHA gels.Conclusion The CHA-gel prepared under optimal reaction conditions have different physical and chemical properties, which set foundation for developing double cross-linked gel with both excellent stability and Enzyme-resistant properties.

Purpose To research out a suitable method for the determination of enzymatic activity for β-glucan synthase. Methods According to the methods for the determination of ΒGS in current reports, taking Grifola frondosa as material, a suitable method for the determination of enzymatic activity for ΒGS was researched out. ΒGS was isolated from the mycelium of Grifola frondosa. Taking glucose as substrate, ΒGS activity was reflected by the consumption of glucose. Consumption of glucose was measured by 3,5-dinitrosalicylic acid (DNS) method, the standard curve for glucose content measurement was plotted by determining absorbance values of different glucose concentrations at 540nm. One unit of ΒGS corresponds to the amount of enzyme which incorporated 1μg glucose into β-glucan in 1 minute. The optimum pH and temperature of the determination of enzymatic activity for ΒGS was determined respectively by carrying out the enzyme assay at different pH levels and temperatures. Results The method for the determination of enzymatic activity for ΒGS was a suitable, correct and usable method which can be popularized widely. The regression equation of the standard curve for glucose content measurement was y=0.0008x-0.0247, R2=0.9996. The optimum pH of the determination of enzymatic activity for ΒGS was pH=5.0 and the optimum temperature was 15°C. The suitable determination process for β-glucan synthase enzymatic activity is as follows: 1.0ml of enzyme was added in a test tube followed by 1.0 ml glucose (1.0mg/ml), and then the pH level was adjusted to 5.0. The test tube was incubated at 15°C for 10 minutes. Later, DNS reagent (1.5ml) was added to the test tube kept in boiling water for 5 minutes and cooled in water. A blank was prepared (2.0ml distilled water and 1.5ml DNS solution). Each solution was fixed the volume to 25ml, the color intensity was estimated at 540 nm using spectrophotometry. Conclusion A method for the determination of enzymatic activity for β-glucan synthase was researched out by taking Grifola frondosa as material, and it can be widely extended and applied by its characteristics of simple, rapid, high sensitivity and low cost.

Purpose To screen out the optimal conditions of cross-linking reaction of preparing for an injectable cross-linked sodium hyaluronate gel(CHA-gel) with higher resistance to hyaluronidase and research for its viscoelasticity and anti-enzyme degradation properties. Methods The CHA hydrogels were prepared with different molecular weights of PEG as cross-linking agent, such as PEG400, PEG1000, PEG6000, PEG10000, PEG20000. The optimal preparing conditions were determined by single factor test and orthogonal experiment. The Enzyme-resistant degradation properties in vitro of CHA-gels were analysed by carbazole and spectrophotometry. Its viscoelasticity was also compared with natural HA-gel by Stabinger method. Results the results of range analysis and variance analysis show that the pH of CHA solution and the ratio of cross-linking agent to HA were significant factors. The optimal preparing conditions of the parameters are 1.5% of HA, 0.001mol/L NaOH, at 37°C, reacting 4hr and 1:15 PEG20000/HA (g/g). Under these conditions, the CHA-gel has excellent Enzyme-resistant properties, R=85.1%, an high percentage of enzyme-resistant property. Its viscoelasticity can reach 61.3×104mPa.s, three times as much as natural HA-gel. Conclusion The CHA-gel with excellent physicochemical properties can be prepared under the optimal conditions, which can set foundation for developing better mechanical and Enzyme-resistant properties products of CHA-gel.

Single-cell injection after immunocytochemistry is a reliable technique for classifying neurons by their morphological structure and their expression of a particular protein. The aim of the present study was to classify the morphological types of calbindin D28k-immunoreactive retinal ganglion cells in the mouse using single-cell injection after immunocytochemistry, to estimate the density of calbindin D28k-immunoreactive retinal ganglion cells in the mouse retina. Calbindin D28k is an important calcium-binding protein that is widely expressed in the central nervous system. Calbindin D28k-immunoreactive retinal ganglion cells were identified by immunocytochemistry and then iontophoretically injected with the lipophilic dye, DiI. Subsequently, the injected cells were imaged by confocal microscopy to classify calbindin D28k-immunoreactive retinal ganglion cells based on their dendritic ramification depth within the inner plexiform layer, field size, and morphology. The cells were heterogeneous in morphology: monostratified or bistratified, with small to large dendritic field size and sparse to dense dendritic arbors. At least 10 different morphological types (CB1–CB10) of calbindin D28k-immunoreactive retinal ganglion cells were found in the mouse retina. The density of each cell type was quite variable (1.98–23.76%). The density of calbindin D28k-immunoreactive cells in the ganglion cell layer of the mouse retina was 562 cells/mm2, 8.18% of calbindin D28k-immunoreactive cells were axon-less displaced amacrine cells, 91.82% were retinal ganglion cells, and approximately 18.17% of mouse retinal ganglion cells expressed calbindin D28k. The selective expression of calbindin D28k in cells with different morphologies may provide important data for further physiological studies of the mouse retina.

It is a common misconception that bats are blind, and various studies have suggested that bats have visual abilities. The purpose of this study was to investigate the cytoarchitecture of calbindin D28K (CB)-, calretinin (CR)-, and parvalbumin (PV)-immunoreactive (IR) neurons in the bat visual cortex using immunocytochemistry. The highest density of CB- and PV-IR neurons was located in layer IV of the visual cortex. The majority of CB- and PV-IR neurons were characterized by a stellate or round/oval shape. CR-IR neurons were predominantly located in layers II/III, and the cells were principally round/oval in shape. Two-color immunofluorescence revealed that 65.96%, 24.24%, and 77.00% of the CB-, CR-, and PV-IR neurons, respectively, contained gamma-aminobutyric acid (GABA). We observed calcium-binding protein (CBP)-IR neurons in specific layers of the bat visual cortex and in specific cell types. Many of the CBP-IR neurons were GABAergic interneurons. These data provide useful clues to aid in understanding the functional aspects of the bat visual system.

To express recombinant structural protein VP1 of enterovirus 71 (EV71) in E.coli and prepare anti-VP1 monoclonal antibodies (mAbs).With P1 gene as a template, EV71 VP1 gene was amplified by PCR and cloned into the expression vector pET-32a(+). After transformation into E.coli TG1, the positive clones were screened and sequenced. The recombinant plasmid was then transformed into BLgold (DE3) and the recombinant protein in inclusion bodies was induced by IPTG and detected by SDS-PAGE. After the inclusion bodies were solubilized with 6 mol/L guanidine hydrochloride, we purified VP1 protein using Ni-NTA affinity chromatography, and then immunized the mice with it to prepare the mAbs against VP1 protein.Recombinant VP1 protein was expressed in E.coli. We obtained totally 24 VP1 monoclonal hybridoma cell strains, of which one was EV71 positive determined by Western blotting and five were positive for IFA.These mAbs are valuable reagents for the development of new vaccines and detection kits for EV71.