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Wang X.,University of Jinan | Huang J.,University of Jinan | Han J.,Shandong Medicinal Biotechnology Center | Yang M.,Shandong Medicinal Biotechnology Center | And 2 more authors.
Frontiers of Optoelectronics | Year: 2014

With the development of biophotonics, biophoton detection technology has been appropriately used. In this paper, the main features and fundamental conceptions of biophotonics were introduced basically. Then the coherence theory of biophoton emission was reviewed. Furthermore, based on this coherence concept, the quantum theory of traditional Chinese medicine (TCM) and properties of Chinese medicinal herbs were presented. To show the nature of biophoton emission in living systems and clarify its basic detection mechanism, high sensitive detection system which allows non-invasive and non-destructive (or less) recording was finally presented. © 2013 Higher Education Press and Springer-Verlag Berlin Heidelberg.


Pang X.,University of Jinan | Pan J.,Shandong Medicinal Biotechnology Center | Gao P.,University of Jinan | Wang Y.,University of Jinan | And 3 more authors.
Biosensors and Bioelectronics | Year: 2015

It was reported that Proprotein convertase subtilisin/kexin type 6 (PCSK6) can promote the progression of rheumatoid arthritis to a higher aggressive status. In this work, a novel visible light induced photoelectrochemical (PEC) platform was designed to detect PCSK6 gene. ZnOatCdTe nanocable arrays/carboxylated g-C3N4 used as the PEC signal generator. Hexagonal ZnO nanorods grew on ITO electrode firstly. CdTe were then electrodeposited on the ZnO nanorods surface to enhance the photogenerated h+/e- separation efficiency. Carboxylated g-C3N4 was utilized to improve h+/e- separation efficiency and anchor the capture probes of PCSK6 gene by the covalent bonding effect. The 5' and 3' primers captured PCSK6 ssDNA by the specific recognition. The linear range was 10pg/mL to 20.0ng/mL with a detection limit of 2pg/mL. © 2015 Elsevier B.V.


Pang X.,University of Jinan | Pan J.,Shandong Medicinal Biotechnology Center | Wang L.,Shandong Medicinal Biotechnology Center | Ren W.,University of Jinan | And 3 more authors.
Biosensors and Bioelectronics | Year: 2015

Proprotein convertase subtilisin/kexin type 6 (PCSK6) plays a major role in promoting the progression of rheumatoid arthritis to a higher aggressive status. A novel highly sensitive photoelectrochemical platform was developed for the detection of PCSK6 by using CdSe quantum dots (QDs)-functionalized TiO2 nanoparticles (NPs) nanohybrids (TiO2@CdSe) as the photo-to-electron conversion medium. TiO2@CdSe showed excellent visible-light absorbency, and much higher photoelectrochemical activity than both CdSe QDs and TiO2 NPs. The 5' and 3' primers of PCSK6 ssDNA acted as capture probes to realize the detection of PCSK6 ssDNA by the specific recognition. The capture probes can be fixed by poly-l-lysine (PLL) through positively strong electrostatic attraction and the carboxyl group of TiO2@CdSe nanohybrids. PLL was electropolymerized on ITO electrode by cyclic voltammetry (CV). Simultaneously, the amino group of PLL can interact with the carboxyl group of TiO2@CdSe nanohybrids to enhance the stability of the photoelectrochemical signal. The fabricated aptsensor exhibited excellent performance towards PCSK6 with a wide linear range (0.5pg/mL to 80.0ng/mL) and a detection limit of 0.1fg/mL. This work opens up a new detection platform for PCSK6 with good sensitivity, reproducibility and stability. © 2015 Elsevier B.V.


PubMed | Shandong Medicinal Biotechnology Center, Shandong Academy of Sciences, Shandong University and University of Jinan
Type: | Journal: Biosensors & bioelectronics | Year: 2015

Based on ZnO nanorods (NRs)/CH3NH3PbI3/nitrogen-doped carbon quantum dots (NCQDs) nanocomposites, the highly sensitive detection of fibroblast-like synoviocyte (FLS) cell was realized by a photoelectrochemical (PEC) biosensor. ZnO/CH3NH3PbI3/NCQDs nanocomposites were exploited as the photo-to-electron generator to produce the signal. CH3NH3PbI3 was spin-coated on ZnO surface after ZnO NRs grew on ITO electrode then by dropping on the modified electrode, NCQDs were diffused and adhered to the surface of ZnO and CH3NH3PbI3. In the presence of EDC/NHS, the combination of CH3NH3PbI3 and NCQDs was achieved by the carboxyl groups (-COOH) and amino groups (-NH2) in the preparation process. Furthermore, the capture probe of FLS cell, CD95 antibody, can be anchored by -COOH and -NH2 groups through EDC/NHS. The specific recognition between the antibody capture probes and cell targets gained high-sensitive detection for FLS cell for the first time. The developed biosensor showed a wide linear range from 1.0 10(4)cell/mL to 10 cell/mL and a low detection limit of 2 cell/mL. This kind of biosensor would provide a novel detection strategy for FLS cell.


PubMed | Shandong Medicinal Biotechnology Center and University of Jinan
Type: | Journal: Biosensors & bioelectronics | Year: 2015

Proprotein convertase subtilisin/kexin type 6 (PCSK6) plays a major role in promoting the progression of rheumatoid arthritis to a higher aggressive status. A novel highly sensitive photoelectrochemical platform was developed for the detection of PCSK6 by using CdSe quantum dots (QDs)-functionalized TiO2 nanoparticles (NPs) nanohybrids (TiO2@CdSe) as the photo-to-electron conversion medium. TiO2@CdSe showed excellent visible-light absorbency, and much higher photoelectrochemical activity than both CdSe QDs and TiO2 NPs. The 5 and 3 primers of PCSK6 ssDNA acted as capture probes to realize the detection of PCSK6 ssDNA by the specific recognition. The capture probes can be fixed by poly-l-lysine (PLL) through positively strong electrostatic attraction and the carboxyl group of TiO2@CdSe nanohybrids. PLL was electropolymerized on ITO electrode by cyclic voltammetry (CV). Simultaneously, the amino group of PLL can interact with the carboxyl group of TiO2@CdSe nanohybrids to enhance the stability of the photoelectrochemical signal. The fabricated aptsensor exhibited excellent performance towards PCSK6 with a wide linear range (0.5 pg/mL to 80.0 ng/mL) and a detection limit of 0.1 fg/mL. This work opens up a new detection platform for PCSK6 with good sensitivity, reproducibility and stability.


Yang Y.,Shandong Medicinal Biotechnology Center | Fu J.-F.,Shandong Medicinal Biotechnology Center | Xu X.-B.,Shandong Medicinal Biotechnology Center | Wang S.-L.,Shandong Medicinal Biotechnology Center
Chinese Journal of Biologicals | Year: 2015

Objective: To express gonadotropin releasing hormone (GnRH) analogue in prokaryotie cells and purify the expressed product. Methods: The 8GnRH analogue was synthesized, based on which the recombinant plasmid containing 16GnRH analogue tandem repeats was constructed by isocaudamer technology. The 8GnRH and 16GnRH analogues were inserted into prokaryotic expression vector pGEX-2t, and the constructed recombinant plasmids were transformed to E. coli BL21. The recombinant 8GnRH analogue or 16GnRH analogue expressed after induction with IPTG under the optimized condition was purified by Glutathione Sepharose 4B column chromatography, and identified by SDS-PAGE and Western blot. Results: Both restriction analysis and sequencing proved that recombinant plasmids pGEX-2t-8GnRH analogue and pGEX-2t-16GnRH analogue were constructed correctly. The expression level of target protein in supernatant reached a peak value after induction with IPTG at a final concentration of 0. 2 mmol/L at 37 °C for 2 h. The expressed recombinant protein, with a relative molecular mass of about 37 000, showed specific binding to rabbit monoclonal antibody against GnRH. The relative molecular mass and concentration of purified recombinant protein were about 1 400 and 260 μg/ml respectively. Conclusion: Recombinant plasmid with multicopy GnRH analogue gene was successfully constructed and highly expressed in E. coli, which laid a foundation of production of GnRH analogue by multicopy method.


Wu X.-Y.,Shandong Medicinal Biotechnology Center | Wang S.-L.,Shandong Medicinal Biotechnology Center
Chinese Journal of Biologicals | Year: 2011

Objective: To develop a novel method for harvest of hematopoietic stem cells and provide an experimental basis for clinical application. Methods: Gelatin sponges (GS) were implanted into the spatium intermusculare of mice hind limbs, in which the migrating cells were isolated 12 d later. The expressions of CD34 +, Sca-1 +, and CD34 +Sca-1 + cells in migrating cells (MCs) and bone marrow cells (BMCs) were determined by flow cytometry and IFA, and the density of hematopoietic colonies by colony-forming cell assay in vitro. Results: The count of migrating cells reached the maximum 12 d after transplantation. The proportions of CD34 +, Sca-1 + and CD34 +Sca-1 + cells (each P < 0.05) as well as the density of hematopoietic colonies were significantly higher in MCs than in BMCs (P < 0.01). Conclusion: An effective method for harvest of hematopoietic stem cells was successfully developed with GS as a biomaterial, by which autologous cell therapy might be feasible.


Wang F.,Shandong Medicinal Biotechnology Center | Wang L.,Shandong Medicinal Biotechnology Center | Jiang H.,Shandong Medicinal Biotechnology Center | Chang X.,Shandong Qianfoshan Hospital | Pan J.,Shandong Medicinal Biotechnology Center
Journal of Rheumatology | Year: 2015

To assess the effect of proprotein convertase subtilisin/kexin type 6 (PCSK6) in the synovial fibroblasts of rheumatoid arthritis (RA). PCSK6 is a proteinase implicated in the proteolytic activity of various precursor proteins and involved in the regulation of protein maturation. Methods. PCSK6 expression was detected in the synovial tissue of 10 patients with RA, 10 controls with osteoarthritis, and 10 controls with ankylosing spondylitis using Western blotting and quantitative real-time PCR. Genotyping of 67 tag single-nucleotide polymorphisms (SNP) was performed using an Illumina VeraCode (Illumina) microarray in a case-control study including 267 patients with RA and 160 healthy controls. Genotyping of 4 other tag SNP was performed using a TaqMan probe genotyping assay in 1056 healthy controls and 1151 patients with RA. Cultured RA synovial fibroblasts (RASF) were transfected with PCSK6 small interfering RNA to study changes in the proliferation, invasion, migration capacity, secretion of inflammatory cytokines, cell cycle, and expression profiles of the RASF. Results. Expression of PCSK6 mRNA and protein was significantly higher in the synovial tissues of individuals with RA than in control tissues. One SNP, rs8029797, was significantly associated with RA (p = 0.011). Knockdown of PCSK6 by RNA interference significantly decreased proliferation, invasion, and migration of RASF. These changes in RASF appeared to be related to reduced tumor necrosis factor-α secretion, G0/G1 arrest, and altered expression of various proteins including those involved in angiogenesis (matrix metalloproteinase 9, nitric oxide synthase trafficking), hypoxia (hypoxia-inducible factor-α, thioredoxin domain containing 5), proliferation (chromosome 10 open reading frame 116), and inflammation [CCL7, chemokine (C-X-C motif) ligand 9, interleukin 26]. Conclusion. PCSK6 is upregulated in the synovial tissues of patients with RA and has a genetic effect on the risk of RA. Inhibition of PCSK6 may play a protective role in the development of RA © 2015. All rights reserved.


Cai W.-W.,Shandong Medicinal Biotechnology Center
Chinese Journal of Biologicals | Year: 2013

Objective: To investigate the effect of various dosages of Antarctic krill powder, alone or in combination with calcium, on osteoporosis in rats. Methods: One hundred of eight week old rats were divided into eight test groups as well as one sham operation control group and one model control group, 10 for each. Osteoporosis model was established by ovariectomy, while the partial fatty tissues adjacent to ovaries of rats in sham operation control group were resected. The rats in OVX + LK, OVX + MK and OVX + HK groups were treated with Antarctic krill powder at low, moderate and high dosages, while those in OVX + Ca group with calcium, those in OVX + LK + Ca, OVX + MK + Ca and OVX + HK + Ca groups with Antarctic krill powder at low, moderate and high dosages + calcium, and those in NaF group with sodium fluoride, by gavage, respectively. However, the rats in sham operation control (Sham) and model control (OVX) groups were fed with normal forage. Twelve weeks later, the serum samples were collected and determined for calcium ion concentration and alkaline phosphatase (ALP) activity. The thigh-bones of rats were collected, observed for differences in bone mineral density and bone maximum load, and determined for ash content, while the shin bone were determined for fluorine content. Results: The serum calcium contents of rats decreased gradually with the increasing dosage of Antarctic krill powder, which showed significant difference in OVX + HK and OVX groups(P < 0.05), while were higher in OVX + LK + Ca, OVX + MK + Ca and OVX + HK + Ca groups than in OVX group. The ALP activity increased with the increasing dosage of Antarctic krill powder, which were higher in OVX + LK, OVX + MK and OVX + HK groups than in OVX group (P < 0.05). The bone density of rats in OVX + LK + Ca, OVX + MK + Ca and OVX + HK + Ca groups were higher than those in OVX, OVX + LK, OVX + MK and OVX + HK groups. The bone maximum loads of rats in OVX + LK, OVX + MK + Ca and OVX + HK + Ca groups were significantly higher than those in OVX group (each P < 0.05). As compared with those in OVX group, the fluorine contents in bones of rats in OVX + LK, OVX + MK and OVX + HK groups increased with the dosage of Antarctic krill powder, while decreased significantly in OVX + LK + Ca, OVX + MK + Ca and OVX + HK + Ca groups (each P < 0.01). The fluorine contents in OVX + LK + Ca, OVX + MK + Ca and OVX + HK + Ca groups were lower than those in OVX + LK, OVX + MK and OVX + HK groups, respectively. The ash content in OVX + LK, OVX + MK and OVX + HK groups were significantly higher than those in OVX group (P > 0.05). Conclusion: The Antarctic krill powder at a low dosage showed satisfactory curative effect on osteoporosis. However, the curative effect of Antarctic krill powder at a moderate dosage was satisfactory when its was administered in combination with calcium.


PubMed | Shandong Medicinal Biotechnology Center
Type: Journal Article | Journal: Molecular medicine reports | Year: 2015

The aim of the present study was to assess the effects of pro-protein convertase subtilisin/kexin type 6 (PCSK6), a proteinase implicated in the proteolytic activity of various precursor proteins and involved in the regulation of protein maturation, in fibroblastlike synoviocytes (FLS) of a rat model of collageninduced arthritis (CIA). Cultured FLS from CIA models were subjected to small interfering RNA mediated PCSK6 knockdown, followed by assessment of the proliferation, invasive and migratory capacity, the secretion of inflammation factors and the cell cycle. Expression of genes associated with proliferation, invasion, migration and inflammation was detected by reverse transcription polymerase chain reaction. The results showed that PCSK6 knockdown significantly decreased the cell proliferation, invasion and migration of FLS from rats with CIA. ELISA showed an obvious decrease of tumor necrosis factor and interleukin 1 secretion, and flow cytometric analysis revealed G0/G1 arrest of FLS following PCSK6 knockdown. Furthermore, a decrease in the mRNA levels of inflammationassociated chemokine CXCL9, angiogenesisassociated genes MMP2, MMP9 and NOSTRIN, hypoxiaassociated gene HIF1, adhesionassociated gene MPZL2, proliferationassociated gene IGF2 and citrullinationassociated gene PADI4 was detected after PCSK6 knockdown. The results of the present study indicated that inhibition of PCSK6 may have a protective role against synovitis in rheumatoid arthritis.

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