Analysis of full and empty ratio of EV71 virus by using capillary zone electrophoresis.

Hand, foot, and mouth disease is a serious public health problem, and the main pathogen is enterovirus 71 (EV71). Its capsid assembly mechanism including capsid protein processing has been widely studied. Full and empty capsids have different immunological efficacy. Therefore, tracking full/empty capsid ratio throughout the EV71 production process is important to ensure consistent product quality and proper dosing response. The analysis of full/empty capsid ratio of intact virus has been widely reported as well. A variety of techniques have been employed to evaluate the full/empty capsid ratios. However, there has not been a rapid, reproducible, and robust assay to determine the full/empty capsid ratios of final and in-process products. In this study, a novel assay based on capillary zone electrophoresis was established. The separation of full and empty species could be achieved within 10 min and the ratio of peak areas was used to calculate the full/empty capsid ratio directly. The results showed good reproducibility and linearity for the determination of full/empty capsid ratios.

Sequential posterior interosseous artery perforator flap for reconstruction of dorsal hand defects.

Dorsal hand defects accompanied by exposure of bones and tendons remain a huge challenge for plastic surgeons. The pedicled perforator flaps have unique advantages in resurfacing the defects. This study aimed to investigate the clinical efficacy of the sequential posterior interosseous artery perforator flap for repairing dorsal hand defects. This study was composed of an anatomical study and clinical application. Anatomically, 30 adult upper limb specimens injected with red latex were dissected, the perforators from the branches of the posterior interosseous artery were observed in the dorsal forearm, and the sequential flap based on them was designed based on the anatomical characteristics. Clinically, nine cases of soft tissue defects on the dorsum of the hand were treated by this flap. Anatomically, the posterior interosseous artery divided into an ascending branch and a descending branch, and the descending branch traveled 1.0 ± 0.3 cm down to divide into the ulnar and radial terminal branches. The ulnar terminal branch presented constantly, and the radial terminal branch had an occurrence rate of 93.33%. Clinically, all flaps survived completely and possessed a soft texture and satisfactory appearance, as well as a nonbloated pedicle, and the donor region had a natural color and appearance with only a linear scar left behind. The sequential posterior interosseous artery perforator flap could become a useful option for repairing dorsal hand defects, as it has the advantages of being easy to perform without sacrificing the main vessels and avoiding donor area skin grafting.

Bilateral anterior elevation prosthesis boosts chondrocytes proliferation in mice mandibular condyle.

OBJECTIVE: We aimed to develop a mouse model predominating in a proliferative response in the articular cartilage of the temporomandibular joints. MATERIALS AND METHODS: Bilateral anterior elevation of occlusion was developed by installing metal tubes onto the incisors of mice with edge-to-edge relation to prevent tooth wear, leading to an increase in the vertical height of the dental occlusion with time. Morphological changes and expression changes in Cyclin D1, Aggrecan, and type II and type X collagen in the mandibular condylar cartilage were detected. In addition, cells were isolated from the mandibular condylar cartilage and exposed to cyclic tensile strain (CTS). RESULTS: Compared with age-matched controls, the tooth length was longer at 3 weeks, 7 weeks, and 11 weeks in BAE mice (p < 0.05), with increased condylar cartilage thickness, matrix amount, and cell number (p < 0.05). Compared with the deep zone cells, CTS stimulated the superficial zone cells to express a higher level of proliferating cell nuclear antigen, Cyclin D1, Aggrecan, and type II collagen but a lower level of type X collagen and alkaline phosphatase. CONCLUSION: Bilateral anterior elevation stimulated the proliferative response in the mandibular condylar cartilage, offering a new therapeutic strategy for cartilage degeneration.

Early growth response 1 reduction in peripheral blood involving condylar subchondral bone loss.

OBJECTIVES: To detect whether early growth response 1 (EGR1) in peripheral blood leucocytes (PBLs) indicates temporomandibular joint (TMJ) osteoarthritis (OA) lesions. MATERIALS AND METHODS: Egr1 mRNA expression levels in PBLs were detected in eight malocclusion patients without temporomandibular disorder (TMD) signs and 16 malocclusion patients with clinical TMD signs with (eight) or without (eight) imaging signs of TMJ OA. Twelve 6-week-old rats were randomized to a control group and a unilateral anterior crossbite (UAC) group and were sampled at 4 weeks. The Egr1 mRNA expression levels in PBLs and protein expression levels in different orofacial tissues were measured. RESULTS: Patients with TMD signs with/without TMJ OA diagnosis showed lower Egr1 mRNA expression levels in PBLs than patients without TMD signs. The lower Egr1 mRNA expression was also found in the PBLs of UAC rats, which were induced to exhibit early histo-morphological signs of TMJ OA lesions. In subchondral bone of UAC rats, EGR1 protein expression was decreased, co-localization of EGR1 with osterix or dentin matrix protein-1 was identified, and the number of EGR1 and osterix double-positive cells was reduced (all p < .05). CONCLUSION: Egr1 reduction in PBLs potentially indicates subchondral bone OA lesions at an early stage.

Catabolic changes of rat temporomandibular joint discs induced by unilateral anterior crossbite.

BACKGROUND: The temporomandibular joint (TMJ) disc plays a role in joint movement and in load absorbance and distribution. An experimental unilateral anterior crossbite (UAC) prosthesis induces mandibular condylar cartilage degeneration in rats. However, the changes in the articular disc are still unknown. OBJECTIVE: To describe changes in the TMJ discs of UAC rats. METHODS: The discs of fifty-four Sprague-Dawley rats, equally distributed into a UAC group and an age-matched sham-operated control group at 4, 12 and 20 weeks (n = 9), were evaluated by gross and histomorphological observation and by detection at the mRNA or protein expression levels of the markers related to the matrix elements. RESULTS: No macro- or micro-morphological differences were observed between groups. However, there were catabolic degradative changes at the molecular level in the UAC group, showing a significant reduction in the mRNA and/or protein expression levels of many molecules. The reduction became worse with time (P < 0.05). The reduced molecules included: (a) those related to the extracellular matrix, such as type I collagen, decorin and fibromodulin; (b) those related to chondrogenesis, such as type II collagen and aggrecan; and (c) those related to osteogenesis, such as alkaline phosphatase and runt-related transcription factor 2. The mRNA expression of vascular endothelial growth factor did not change. In contrast, fibronectin, which can promote wound healing, and its N-terminal fragment, which can induce cartilage degradation, were accumulated (P < 0.05). CONCLUSION: TMJ discs were stimulated to catabolic changes by the aberrant dental occlusion and seemed to go to inanimate with time.

Molecular changes in peripheral blood involving osteoarthritic joint remodelling.

Biomarkers of temporomandibular joint (TMJ) osteoarthritis (OA) remain unknown. The objective was to detect whether molecular biomarkers from peripheral blood leucocytes (PBLs) engage in TMJ OA lesions. Thirty-four six-week-old Sprague Dawley rats were used. The top upregulated gene ontology categories and gene-fold changes in PBLs were detected by a microarray analysis comparing rats that received 20-week unilateral anterior crossbite (UAC) treatment with age-matched controls (n = 4). Twenty weeks of UAC treatment had been reported to induce TMJ OA-like lesions. The other twenty-four rats were randomly placed in the UAC and control groups at 12- and 20-week time points (n = 6). The mRNA expression levels of the selected biomarkers derived from the microarray analysis and their protein expression in the alveolar bone and TMJ were detected. The microarray analysis indicated that the three most highly involved genes in PBLs were Egr1, Ephx1 and Il10, which were confirmed by real-time PCR detection. The increased protein expression levels of the three detected molecules were demonstrated in cartilage and subchondral bone (P < 0.05), and increased levels of EPHX1 were reported in discs (P < 0.05); however, increased levels were not present in the alveolar bone. Immunohistochemistry revealed the increased distribution of EGR1-positive, EXPH1-positive and IL10-positive cells predominantly in the osteochondral interface, with EXPH1 also present in TMJ discs. In conclusion, the increased mRNA expression of Egr1, Ephx1 and Il10 in PBLs may serve as potential biomarkers for developed osteoarthritic lesions relating to osteochondral interface hardness changes induced by dental biomechanical stimulation.

The coating of smart pH-responsive polyelectrolyte brushes in capillary and its application in CE.

A novel pH-responsive coating technique was developed and applied to CE successfully in this paper. The coating was formed by bonding mixed opposite charge poly(acrylic acid) and poly(2-vinylpyridine) randomly onto the inner wall of a silica capillary. The coating processes were first characterized by ellipsometry and atomic force microscopy at macroscale and microscale, respectively. Measurements of EOF were implemented to confirm the coating. Direction and velocity of EOF became controllable from negative to positive, showing a perfect sigmoidal curve as the coating net charges alternated by the pH of BGE. The control of the EOF makes it possible to analyze different kinds of small molecules, peptides, and proteins successfully in the same capillary. Results showed that the stability and reproducibility for separations of fluoroquinolone standards were satisfactory for more than a hundred separations. A series of basic and acidic protein standards were separated with admirable efficiency and minimal adsorption using both polarities. The separation of tryptic BSA digest showed that the prepared capillary has immense potential in analyzing a single sample with both acidic and basic separations, which achieved the expectation in proteomics study by CE-MS.

Synergistic Effects of Quercetin-Modified Silicone Gel Sheet in Scar Treatment.

Both silicone gel and quercetin are effective in scar treatment but have different action mechanisms. Quercetin is mainly applied in the gel form and can lead to poor adhesion of silicone gel sheet; therefore, they cannot be combined in clinical use. In this study, a silicone gel sheet that releases quercetin in a sustained manner for 48 hours was successfully developed. Four round scars (Ø: 1 cm) were made in the ears of New Zealand albino rabbits (n = 10). After scar healing, the rabbits were divided into four groups: blank control group with no treatment, silicone gel sheet group with dressing change every 2 days, quercetin group with dressing change three times daily, and combination treatment group with dressing change every 2 days. Scar assessment was performed 3 months later. Transepidermal water loss showed no difference between the combination treatment group and the silicone gel sheet group, but was lower than that in the quercetin group and the blank control group. Immunohistochemistry of CD 31 and proliferating cell nuclear antigen showed the following results: combination treatment group < silicone gel sheet group = quercetin group < blank control group. Polymerase chain reaction results showed that the expression of type-I and type-III collagen in the combination treatment group and the quercetin group was significantly lower than that in the other two groups. Thus, quercetin-modified silicone gel sheet combines the advantages of the two treatments and is more effective at inhibiting cell proliferation in scar tissue than either of the two treatments alone.

[Application of Vectra 3D facial imaging technology in orthodontic teaching program].

PURPOSE: To evaluate the effectiveness of Vectra 3D facial imaging technology in enhancing orthodontic teaching and training efficiency. METHODS: Thirty-two dental students, enrolled in 2014 as five-year and eight-year curriculum in School of Somatology, Air Force Military Medical University were selected as the subjects of this research. As an important teaching facility for training the students to practice orthodontic clinical examinations, 2D and 3D facial imaging systems were selected in this study for the students to collect and analyze the data relating to the facial complexion and appearance. The students were at first instructed to use the traditional 2D facial imaging system for 20 minutes, and then Vectra 3D facial imaging system for another 20 minutes. The students were required to deliberate on the specifically designed questionnaires to input their own assessment on these two modalities. The outcomes were quantitatively analyzed using SPSS 24.0 software package. RESULTS: As to the items 2, 5, 6 and 7 in the questionnaire, which indicated the following queries respectively: whether the use of imaging systems could inspire students' learning interest, whether the results drawn from these two imaging facial systems were accurate and reliable, whether the subjective bias were trivial , and whether these two imaging systems were feasible for orthodontic treatment appraisals. The results showed that there were significant differences in these four items between the two groups（P<0.05). As to the Items 1, 3 and 4 , namely, whether the acquisition of the teaching materials was an easy access, whether these two facial analytical regimes were beneficial for the students to obtain the new knowledge, and whether this specific teaching facility was easy for the students to manage, there was no significant difference between the two groups (P>0.05). CONCLUSIONS: Served as a new teaching syllabus facilitation, Vectra three-dimensional facial imaging system demonstrates a more satisfactory impetus for the students to learn than the traditional two-dimensional imaging system. Pragmatically, the analytical data resulted from the former remains more accountable than that of the latter.

Masseter response to long-term experimentally induced anterior crossbite in Sprague-Dawley rats.

OBJECTIVE: To detect the long-term response to unilateral anterior crossbite (UAC) in masticatory muscles and in molecular biomarkers of peripheral blood leukocytes. DESIGN: Fifty-six six-week-old Sprague-Dawley rats were used. The gene-fold changes in peripheral blood leukocytes were detected by the microarray analysis to compare the rats that received 20-week UAC treatment with age-matched controls (n = 4). Muscle atrophy-related gene Fbxo32 was selected based on the data of the microarray analysis verified by using real-time PCR. The remaining 36 rats were randomly separated in the UAC and control groups at 12 and 20 weeks (n = 12). The protein expression of Fbxo32 and the muscle injury and myogenesis-related markers, αB-crystallin and desmin, were detected in the masseter and lateral pterygoid muscles by western blot assay. RESULTS: In the 20-week UAC group, the masseter muscle weight was lower than that in the age-matched control group, and the expression level of Fbxo32 gene in peripheral blood leukocytes was increased according to the microarray analysis confirmed by real-time PCR detection. The increased protein expression levels of Fbxo32 were detected in the masseter in the 20-week UAC group, and the protein expression levels of desmin and αB-crystallin were decreased at this time point. No similar changes were detected in the lateral pterygoid muscle. CONCLUSIONS: Masseter atrophy is induced by long-term stimulation of UAC. The increased expression of the Fbxo32 gene in peripheral blood leukocytes may be a candidate biological marker of masseter atrophy.

Dual plastic film and straw mulching boosts wheat productivity and soil quality under the El Nino in semiarid Kenya.

The extreme climate events such as El Nino seriously threaten crop production and agro-ecological sustainability because of the aggravated environmental stresses worldwide, particularly in sub-Saharan Africa. To address this issue, we investigated the effects of dual plastic film and straw mulching in ridge-furrow (RF) system on wheat productivity, soil carbon and nitrogen stocks in a semiarid area in Kenya from 2015 to 2017. The experimental site represents a typical semiarid continental monsoon climate, and soil type is chromic vertisols. Field experiment with randomized block design consisted of six RF treatments as follows: 1) dual black plastic film and straw mulching (RFbS), 2) dual transparent plastic film and straw mulching (RFtS), 3) sole black plastic film mulching (RFb), 4) sole transparent plastic mulching RF (RFt), 5) sole straw mulching (RFS) and 6) no mulching (CK). The results indicated that seasonal dynamics of rainfall and air temperature fit in with the weather type of El Nino over four growing seasons. RFbS, RFtS, RFb and RFt significantly increased soil water storage (SWS), topsoil temperature, aboveground biomass, grain yield and water use efficiency across four growing seasons (p < 0.05) as compared with CK. Among all the treatments, RFbS and RFtS achieved the greatest SWS, AgB, grain yield and WUE, owing to improved soil hydro-thermal status in both treatments. Critically, RFbS and RFtS significantly improved soil organic carbon and total nitrogen, soil bulk density and the C:N ratio following four growing seasons, comparing with other treatments (p < 0.05). Besides, RFbS and RFtS gave the highest economic returns among all treatments. For the first time, we found that dual plastic film and straw mulching could serve as a sustainable land management to boost wheat productivity and improve soil quality under El Nino in semiarid areas of SSA.

Injury responses of Sprague-Dawley rat jaw muscles to an experimental unilateral anterior crossbite prosthesis.

OBJECTIVE: Dental occlusion are frequently changed in clinic. Molecular responses in jaw muscles to aberrant dental occlusion are changes are attractive, yet remain are obscure. DESIGN: Unilateral anterior crossbite (UAC) prostheses were applied to Sprague-Dawley rats and then ceased after two weeks to detect the reactions of the masseter, a representative jaw elevator, and the lateral pterygoid muscle (LPM), a representative jaw depressor. RESULTS: Two weeks of UAC elicited mild injury of the two muscles. Myogenesis and protective reactions were detected as increases in αB-crystallin expression in the masseter after 3 days and in the LPM after 2 weeks, and increases in desmin expression in both muscles after 2 weeks. A switch in fibre types from IIb to IIx occurred in the LPM but not in the masseter. Inflammatory responses, shown by the infiltration of inflammatory cells and increases in TNF-α mRNA expression, and fibrosis responses, shown by increased mRNA expression of Type I and III collagens, appeared very mild in the two muscles. These responses were partially recovered by the cessation of UAC. During the whole process, no obvious changes were observed in mitochondrial function, as indicated by the levels of proliferator-activated receptor γ coactivator 1α, mitofusin-2 and voltage-dependent anion channel. CONCLUSIONS: UAC causes injury and very limited inflammatory and fibrosis adaption in the masseter and LPM. Both muscles respond with myogenesis and protective activity. The LPM responds also with muscle fibre isoform alternations. These alterations were partially recovered by the cessation of dental stimulation at an early stage.

[Anatomical basis of the flap based on the perforator of the deep palmar arch].

Objective: To investigate the morphological characters of the dorsal perforators originated from the deep palmar arch,so as to provide anatomic basis for V-Y advanced perforator flap. Methods: The following contents were investigated in 30 aduh hand specimens perfused with red latex under surgical magnifier:① The origin, courses, branches and distribution of the dorsal perforators originated from the deep palmar arch. ②The characters of anastomosis among the dorsal perforators, the dorsal carpal and metacarpal arteries. Mimic operation was performed on another fresh specimens perfused with red latex. Results: There were three perforators originated from the deep palmar arch, which passed through the 2nd-4th dorsal interossei and then divided into an ascending branch and a descending branch at the dorsum of hand. Then the ascending branch anastomosed with the dorsal carpal artery, and the descending branch stretched to the 2nd-4th dorsal metacarpal arteries. The originating outer diameters of the 1st-3rd perforators were (1.1 ±0.2) mm,(0.9 ±0.3) mm and (0.7 ±0.1) mm respectively, and the length of the stems were (1.1 ±0.3) cm,(1.0 ±0.2)cm and (0.9 ±0.1) cm respectively. Conclusions: The V-Y advanced perforator flap with the dorsal perforator of the deep palmar arch as its vascular pedicle could be used to repair the dorsal carpal or dorsal metacarpal soft tissue defects.

Activation of α2A-adrenergic signal transduction in chondrocytes promotes degenerative remodelling of temporomandibular joint.

This study tested whether activation of adrenoreceptors in chondrocytes has roles in degenerative remodelling of temporomandibular joint (TMJ) and to determine associated mechanisms. Unilateral anterior crossbite (UAC) was established to induce TMJ degeneration in rats. Saline vehicle, α2- and ß-adrenoreceptor antagonists or agonists were injected locally into the TMJ area of UAC rats. Cartilage degeneration, subchondral bone microarchitecture and the expression of adrenoreceptors, aggrecans, matrix metalloproteinases (MMPs) and RANKL by chondrocytes were evaluated. Chondrocytes were stimulated by norepinephrine to investigate signal transduction of adrenoreceptors. Increased α2A-adrenoreceptor expression was observed in condylar cartilage of UAC rats, together with cartilage degeneration and subchondral bone loss. Norepinephrine depresses aggrecans expression but stimulates MMP-3, MMP-13 and RANKL production by chondrocytes through ERK1/2 and PKA pathway; these effects were abolished by an α2A-adrenoreceptor antagonist. Furthermore, inhibition of α2A-adrenoreceptor attenuated degenerative remodelling in the condylar cartilage and subchondral bone, as revealed by increased cartilage thickness, proteoglycans and aggrecan expression, and decreased MMP-3, MMP-13 and RANKL expressions in cartilage, increased BMD, BV/TV, and decreased Tb.Sp in subchondral bone. Conversely, activation of α2A-adrenoreceptor intensified aforementioned degenerative changes in UAC rats. It is concluded that activation of α2A-adrenergic signal in chondrocytes promotes TMJ degenerative remodelling by chondrocyte-mediated pro-catabolic activities.

Preparation, characterization and adsorption properties of a novel 3-aminopropyltriethoxysilane functionalized sodium alginate porous membrane adsorbent for Cr(III) ions.

In this study, we prepared 3-aminopropyl-triethoxysilane (APTEOS) functionalized sodium alginate (SA) porous membrane adsorbent (APTEOS/SA) and tested its adsorption performance for removing of Cr(III) ions. The physico-chemical properties of the pristine and Cr(III) ions loaded APTEOS/SA were investigated by FT-IR, SEM-EDX, TG, AFM, and contact angle goniometer methods. To investigate the adsorption kinetics of Cr(III) ions onto this newly developed APTEOS/SA, we performed a batch of experiments under different adsorption conditions: solution pH, adsorbent dose, initial Cr(III) ion concentration, adsorption temperature, and contact time. The APTEOS/SA exhibited an encouraging uptake capacity of 90.0mg/g under suitable adsorption conditions. To study the mechanism of adsorption process, we examined the Lagergren pseudo-first-order and pseudo-second-order kinetic model, the intra-particular diffusion model, and the Crank model. Kinetics experiments indicated that the pseudo-first-order model displayed the best correlation with adsorption kinetics data. The Crank model showed that the intra-particle solute diffusion was the main rate-controlling step. Furthermore, our adsorption equilibrium data could be better described by the Freundlich equation. We also carried out consecutive adsorption-desorption experiments eight times to show that the APTEOS/SA has encouraging adsorption-desorption efficiencies. The results indicates that the prepared adsorbent is promising for using as an effective and economical adsorbent for Cr(III) ions removal.

Biocompatibility evaluation of electrospun aligned poly (propylene carbonate) nanofibrous scaffolds with peripheral nerve tissues and cells in vitro.

BACKGROUND: Peripheral nerve regeneration across large gaps is clinically challenging. Scaffold design plays a pivotal role in nerve tissue engineering. Recently, nanofibrous scaffolds have proven a suitable environment for cell attachment and proliferation due to similarities of their physical properties to natural extracellular matrix. Poly(propylene carbonate) (PPC) nanofibrous scaffolds have been investigated for vascular tissue engineering. However, no reports exist of PPC nanofibrous scaffolds for nerve tissue engineering. This study aimed to evaluate the potential role of aligned and random PPC nanofibrous scaffolds as substrates for peripheral nerve tissue and cells in nerve tissue engineering. METHODS: Aligned and random PPC nanofibrous scaffolds were fabricated by electrospinning and their chemical characterization were carried out using scanning electron microscopy (SEM). Dorsal root ganglia (DRG) from Sprague-Dawley rats were cultured on the nanofibrous substrates for 7 days. Neurite outgrowth and Schwann-cell migration from DRG were observed and quantified using immunocytochemistry and SEM. Schwann cells derived from rat sciatic nerves were cultured in electrospun PPC scaffold-extract fluid for 24, 48, 72 hours and 7 days. The viability of Schwann cells was evaluated by 3-[4,5-dimethyl(thiazol-2-yl)-2,5-diphenyl] tetrazolium bromide (MTT) assay. RESULTS: The diameter of aligned and random fibers ranged between 800 nm and 1200 nm, and the thickness of the films was approximately 10 - 20 µm. Quantification of aligned fiber films revealed approximately 90% alignment of all fibers along the longitudinal axis. However, with random fiber films, the alignment of fibers was random through all angle bins. Rat DRG explants were grown on PPC nanofiber films for up to 1 week. On the aligned fiber films, the majority of neurite outgrowth and Schwann cell migration from the DRG extended unidirectionally, parallel to the aligned fibers. However, on the random fiber films, neurite outgrowth and Schwann cell migration were randomly distributed. A comparison of cumulative neurite lengths from cultured DRGs indicated that neurites grew faster on aligned PPC films ((2537.6 ± 987.3) µm) than randomly-distributed fibers ((493.5 ± 50.6) µm). The average distance of Schwann cell migration on aligned PPC nanofibrous films ((2803.5 ± 943.6) µm) were significantly greater than those on random fibers ((625.3 ± 47.8) µm). The viability of Schwann cells cultured in aligned PPC scaffold extract fluid was not significantly different from that in the plain DMEM/F12 medium at all time points after seeding. CONCLUSIONS: The aligned PPC nanofibrous film, but not the randomly-oriented fibers, significantly enhanced peripheral nerve regeneration in vitro, indicating the substantial role of topographical cues in stimulating endogenous nerve repair mechanisms. Aligned PPC nanofibrous scaffolds may be a promising biomaterial for nerve regeneration.

Immunoaffinity extraction of testosterone by antibody immobilized monolithic capillary with on-line laser-induced fluorescence detection.

An immunoaffinity capillary column has been made with poly (2-vinyl-4, 4-dimethylazlactone-co-2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (VDA-co-HEMA-co-EDMA) monolith as a support for the immobilized antibody. The monolith is prepared by UV-initiated in situ polymerization, followed by immobilizing anti-testosterone polyclonal antibody through the rapid reaction with VDA. Fluorescence labeled testosterone at C(3) is designed as a tracer to estimate the extraction ability of this immunoaffinity column, and to optimize the immunoextraction process, such as washing, eluting, incubation and injection. The performance in a more realistic application is then demonstrated successfully for the rapid extraction of testosterone (T) by competitive immunoassay and on-line laser-induced fluorescence (LIF) detection. This immunoaffinity monolith is easily prepared, with high mechanical strength and low flow resistance. It is a satisfactory material as an immunoextractor to detect small molecular compounds specifically and rapidly with high sensitivity (sub ng/mL level).