Highly Sensitive and Linear Resonator-Based Biosensor for White Blood Cell Counting: Feasible Measurement Method and Intrinsic Mechanism Exploration.

Since different quantities of white blood cells (WBCs) in solution possess an adaptive osmotic pressure of cells, the WBCs themselves and in solution have similar concentrations, resulting in them having similar dielectric properties. Therefore, a microwave sensor could have difficulty in sensing the quantity variation when WBCs are in solution. This paper presents a highly sensitive, linear permittivity-inspired microwave biosensor for WBCs, counting through the evaporation method. Such a measurement method is proposed to record measurements after the cell solution is dripped onto the chip and is completely evaporated naturally. The proposed biosensor consists of an air-bridged asymmetric differential inductor and a centrally located circular fork-finger capacitor fabricated on a GaAs substrate using integrated passive fabrication technology. It is optimized to feature a larger sensitive area and improved Q-factor, which increases the effective area of interaction between cells and the electromagnetic field and facilitates the detection of their changes in number. The sensing relies on the dielectric properties of the cells and the change in the dielectric constant for different concentrations, and the change in resonance properties, which mainly represents the frequency shift, corresponds to the macroscopic change in the concentration of the cells. The microwave biosensors are used to measure biological samples with concentrations ranging from 0.25 × 106 to 8 × 106 cells per mL in a temperature (26.00 ± 0.40 °C) and humidity (54.40 ± 3.90 RH%) environment. The measurement results show a high sensitivity of 25.06 Hz/cells·mL-1 with a highly linear response of r2 = 0.99748. In addition, a mathematical modeling of individual cells in suspension is performed to estimate the dielectric constant of individual cells and further explain the working mechanism of the proposed microwave biosensor.

Cedrol from Ginger Ameliorates Rheumatoid Arthritis via Reducing Inflammation and Selectively Inhibiting JAK3 Phosphorylation.

Ginger, as a food spice, is widely applied due to its extensive effects. Cedrol (CE) found in ginger is a sesquiterpene with anti-inflammatory activity. The objective of this research is to discuss the efficacy of CE on ameliorating rheumatoid arthritis (RA). CE inhibited chronic inflammation and pain in a dose-dependent manner accompanied by rapid onset and long duration. Besides, CE treatment effectively ameliorated the paw edema volume and arthritis score with no significant effect on body weight. Organ index, T-cell and B-cell proliferation, histopathology, and immunohistochemistry demonstrated that CE had immunological enhancement and attenuated RA effects. Remarkably, inhibition of phosphorylated-JAK3 protein, thereby abating the secretion of pro-inflammatory cytokines and inflammation-related mediators, was involved in the potential mechanism of CE efficiency through forming a hydrogen bond with ARG953 and ILE955 in the JAK3 active pocket. At the same time, the pharmacokinetic results showed that the absolute bioavailability of CE at 20, 40, and 80 mg/kg was 30.30, 23.68, and 16.11%, respectively. The current results offered clues for mastering the ameliorated RA of CE and further perfected the effective substance basis on the anti-inflammatory effect of ginger, which was beneficial for further applications.

Cedrol attenuates collagen-induced arthritis in mice and modulates the inflammatory response in LPS-mediated fibroblast-like synoviocytes.

Ginger has been used as a flavoring agent and traditional medicine for a long time in Asian countries. Pharmacological studies showed that it has antiemetic, anti-inflammatory and analgesic effects, which is attributed to its chemical constituents. The aim of the present study is to investigate the anti-rheumatoid arthritis properties of cedrol (CE) found in ginger. In an in vivo anti-RA study, CE remarkably alleviated the paw swelling and arthritis score in CE-treated CIA mice compared with the model group. The neutrophil count and the productions of TNF-α and IL-1ß were inhibited, and the expressions of Rankl, Cox-1 and Cox-2 were down-regulated at the mRNA level. Radiologic evaluation, histopathological analysis and immunohistochemistry indicated that CE ameliorated inflammatory cell infiltration, synovial hyperplasia, and bone and cartilage damage, and exhibited an immunotherapeutic effect. The MTT assay demonstrated that CE (10-10-10-5 M) had no cytotoxicity on fibroblast-like synoviocytes (FLSs), and exhibited an inhibitory effect on the proliferation of LPS-induced FLSs at concentrations of 10-6 M and 10-5 M. Mechanism research showed that the suppressed expressions of pivotal inflammatory mediators including COX-1 and COX-2 subsequently reduced the production of PGE2, thereby causing the secretions of pro-inflammatory cytokines, ultimately attenuating the progression of inflammation. Meanwhile, the reduction in the mRNA levels of Mmp-13 and Mcp-1 responsible for osteoclastogenesis resistance was detected. This illustrated that CE showed anti-rheumatoid arthritis properties via blocking the phosphorylation of ERK/MAPK and p65/NF-κB signaling pathways in LPS-activated FLSs. The current research suggested that CE is an important functional component in ginger, which may be a promising candidate drug for RA therapy.

Seroprevalence and genetic characterization of Toxoplasma gondii in masked palm civet (Paguma larvata) in Hainan province, tropical China.

In the present study, the seroprevalence, risk factors and genotyping of Toxoplasma gondii in masked palm civet were investigated in tropical China. A total of 500 serum were collected from five administrative farms in tropical China, and assayed for T. gondii antibodies by modified agglutination test (MAT). The brain samples of 20 aborted fetuses were examined by semi-nested-PCR, and positive aborted fetuses (50%) were necropsied to collect the brain tissue for molecular and bioassay examinations. Genomic DNA was extracted from the 29 brain tissues of infected mice and T. gondii B1 gene was amplified using multilocus PCR-RFLP. Overall, 27.6% (95% CI: 23.682-31.518) of the animals was positive for T. gondii antibodies. Ages of masked palm civet was considered as a main risk factor associated with T. gondii infection. 4 DNA samples (13.8%) were positive for the T. gondii B1 gene. Three samples belong to ToxoDB#9, and one belongs to genotype the type II variant (ToxoDB genotype#3). Our results indicated that ToxoDB Genotype#9 has a distribution in masked palm civet that could be potential reservoirs for T. gondii transmission, which may pose a threat to human health.

Seroprevalence and risk factors of Chlamydia psittaci infection in domestic geese Anser domestica, in Hainan province of China.

Chlamydia psittaci, the agent of psittacosis in humans, infects a wide range of bird species. To assess the risk of psittacosis posed by domestic geese in China, the seroprevalence of C. psittaci infection in domestic geese in Hainan province, tropical China was examined using indirect hemagglutination assay (IHA). The overall seroprevalence was estimated at 25.6% (461/1800; 95% CI: 23.6-27.6). The C. psittaci seroprevalence ranged from 19% (95% CI: 14.6-23.4) to 34% (95% CI: 28.6-39.4) among different regions in Hainan province, and the differences were statistically significant (P<0.01). The risk factors significantly associated with C. psittaci seroprevalence were the presence of hygiene conditions, age, gender, and environment of geese in the farms. The results of the present investigation indicated the high seroprevalence of C. psittaci infection in geese in Hainan province, tropical China. Close contact with these geese is associated with a risk of zoonotic transmission of C. psittaci. Public education should be implemented to reduce the risk of avian to human transmission of such a pathogenic agent. To the best of our knowledge, this is the first report documenting the occurrence of C. psittaci seroprevalence in geese in China.

Seroprevalence, risk factors and genotyping of Toxoplasma gondii in domestic geese (Anser domestica) in tropical China.

BACKGROUND: Little information is available about the seroprevalence of Toxoplasma gondii infection in geese (Anser domestica) in China. In the present investigation, the seroprevalence, risk factors and genotyping of T. gondii in geese were investigated in Hainan province, tropical China. FINDINGS: A total of 600 serum samples and 150 brain tissue samples were collected from six administrative regions in tropical China, and assayed for T. gondii antibodies by Indirect Haemagglutination (IHA) test. Genomic DNA was extracted from the 30 brain tissues of seropositive geese and T. gondii B1 gene was amplified using a semi-nested PCR. DNA samples giving positive B1 amplification were then genetically characterized using multi-locus PCR-RFLP. Overall, 17% (95% CI: 14-20) of the animals were positive for T. gondii antibodies. Presence of cats in the household (odds ratio, OR 3), hygiene (OR 2.3) and presence of stray cat around the house (OR 2.3) were considered as main risk factors associated with T. gondii infection. Of 30 DNA samples, three were positive for the T. gondii B1 gene, two showed complete genotyping results. Only one genotype (type II) was identified. CONCLUSIONS: The results of the present survey indicated the presence of T. gondii infection in geese in tropical China. Therefore, it is imperative that improved integrated measures be carried out to prevent and control T. gondii infection in geese in this province. This is the first report documenting the occurrence of T. gondii genotype in geese in China.

Seroprevalence and molecular detection of Mycoplasma ovipneumoniae in goats in tropical China.

In the present study, the seroprevalence and genetic identification of Mycoplasma ovipneumoniae infection in goats were investigated in Hainan Province, tropical China between October 2012 and October 2013. A total of 1,210 serum samples collected from 16 herds in various administrative regions in tropical China were evaluated using indirect hemagglutination assay (IHA). Antibodies to M. ovipneumoniae were tested in (31.7 %, 95 % confidence interval (CI) 29-34.3) 383 of 1,210 serum samples (IHA titer ≥1:16). The M. ovipneumoniae seroprevalence ranged from 26.8 % (95 % CI 20.8-32.9) to 39 % (95 % CI 30.8-47.2) among different regions in tropical China, and the difference was statistically significant (P < 0.01). The seroprevalence of M. ovipneumoniae infection in goats was higher in winter (46.1 %, 95 % CI 39.6-52.5) and spring (33.8 %, 95 % CI 28.3-39.3) than in autumn (27.5 %, 95 % CI 22.6-32.3) and summer (24.7 %, 95 % CI 20.3-29.1), and the difference was statistically significant (P < 0.01). In addition, DNA was extracted from nasal swab; lung samples and the 16S rRNA gene sequences were amplified by polymerase chain reaction (PCR) and then sequenced. Twenty-four of 329 (7.3 %) nasal swab samples and 73 of 280 (26.1 %) pneumonic lung tissues were found to contain M. ovipneumoniae, respectively. The results of the present survey indicate that M. ovipneumoniae infection is highly prevalent in goats in tropical China. This is the first report of the comprehensive survey of M. ovipneumoniae prevalence in goats in China.

AD-1, a novel ginsenoside derivative, shows anti-lung cancer activity via activation of p38 MAPK pathway and generation of reactive oxygen species.

BACKGROUND: Ginseng is a traditional Chinese herb that has been used for thousands of years. In the present study, effects and mechanisms of AD-1 were evaluated for its development as a novel anti-lung cancer drug. METHODS: The cytotoxic activity was evaluated by MTT assay. Flow cytometry was employed to detect cell cycle, apoptosis and ROS. Western blot and immunohistochemistry were used to analyze signaling pathways. Lung cancer xenograft models were established by subcutaneous implantation of A549 or H292 cells into nude mice. RESULTS: AD-1 concentration-dependently reduces lung cancer cell viability without affecting normal human lung epithelial cell viability. In A549 and H292 lung cancer cells, AD-1 induces G0/G1 cell cycle arrest, apoptosis and ROS production. The apoptosis can be attenuated by a ROS scavenger - N-acetylcysteine (NAC). In addition, AD-1 up-regulates the expression of p38 and ERK phosphorylation. Addition of a p38 inhibitor SB203580, suppresses the AD-1-induced decrease in cell viability. Furthermore, genetic silencing of p38 attenuates the expression of p38 and decreases the AD-1-induced apoptosis. Treatment with NAC reduces AD-1-induced p38 phosphorylation, which indicates that ROS generation is involved in the AD-1-induced p38 activation. In mice, oral administration of AD-1 (10-40mg/kg) dose-dependently inhibited the growth of xenograft tumors without affecting body weight and decreases the expression of VEGF, MMP-9 and CD34 in tumor tissue. TUNEL staining confirms that the tumors from AD-1 treated mice exhibit a markedly higher apoptotic index. CONCLUSIONS AND GENERAL SIGNIFICANCE: These data support development of AD-1 as a potential agent for lung cancer therapy.

Application of 1-hydroxyethylidene-1, 1-diphosphonic acid in boiler water for industrial boilers.

The primary method used for boiler water treatment is the addition of chemicals to industrial boilers to prevent corrosion and scaling. The static scale inhibition method was used to evaluate the scale inhibition performance of 1-hydroxyethylidene-1, 1-diphosphonic acid (HEDP). Autoclave static experiments were used to study the corrosion inhibition properties of the main material for industrial boilers (20# carbon steel) with an HEDP additive in the industrial boiler water medium. The electrochemical behavior of HEDP on carbon steel corrosion control was investigated using electrochemical impedance spectroscopy and Tafel polarization techniques. Experimental results indicate that HEDP can have a good scale inhibition effect when added at a quantity of 5 to 7 mg/L at a test temperature of not more than 100 °C. To achieve a high scale inhibition rate, the HEDP dosage must be increased when the test temperature exceeds 100 °C. Electrochemical and autoclave static experimental results suggest that HEDP has a good corrosion inhibition effect on 20# carbon steel at a concentration of 25 mg/L. HEDP is an excellent water treatment agent.

Novel dammarane-type sapogenins from Panax ginseng berry and their biological activities.

Three new dammarane-type sapogenins (1, 3, and 5) together with two known ones (2 and 4) were isolated from the total hydrolyzed saponins extracted from Panax ginseng berry. Their structures were elucidated using a combination of 1D and 2D (1)H and (13)C NMR spectra and mass spectroscopy as 20(R)-25-methoxyl-dammarane-3ß,12ß,20-triol (1), 20(R)-25-methoxyl-dammarane-3ß,6α,12ß,20-tetrol (2), 20(R)-20-methoxyl-dammarane-3ß,12ß,25-triol (3), 20(R)-20,25-dimethoxyl-dammarane-3ß,12ß-diol (4), and (12R,20S,24S)-20,24-; 12,24-diepoxy-dammarane-3ß-ol (5). Their antitumor activities were evaluated in six human cancer cell lines. The novel compounds 1 and 3 showed significant cytotoxic activity against the six cell lines. The IC(50) values of 3 against HepG2, Colon205, and HL-60 were the lowest (8.78, 8.64, and 3.98 µM, respectively). Compounds 1 and 20(S)-25-OCH(3)-PPD, which are a pair of configuration isomers, showed a 10- to 100-fold greater growth inhibition than ginsenoside-Rg(3) (an anti-cancer clinical agent in China). The data presented here may be useful for the development of novel anti-cancer agents.

Di-4-pyridyl disulfide-isophthalic acid (1/1).

In the title 1:1 cocrystal, C(10)H(8)N(2)S(2)·C(8)H(6)O(4), the asymmetric unit contains an isophthalic acid mol-ecule and a 4,4'-dipyridyl disulfide mol-ecule. The two carboxyl groups of isophthalic acid inter-act with neighbouring 4,4'-dipyridyl disulfide mol-ecules through O-H⋯N hydrogen bonds, forming a one-dimensional zigzag chain. Neighbouring chains are linked to each other via π-π stacking inter-actions between the pyridyl rings of adjacent 4,4'-dipyridyl disulfide mol-ecules [centroid-centroid distance = 3.7346 (6) Å], resulting in a layered motif. The dihedral angle between pyridine rings of 84.13 (7)° and the C-S-S-C torsion angle of 91.95 (1)° confirm the gauche conformation of 4,4'-dipyridyl disulfide.

[Sorption behavior of bisphenol A on anaerobic sludge].

Batch experiments were conducted to investigate the sorption behavior of bisphenol A (BPA) with low concentration on anaerobic sludge. The autoclaved inactivated sludge was used to minimize the influence of degradation of activated sludge. The results showed that the sorption of BPA on anaerobic sludge was a quick process. Anaerobic sludge had achieved 89.1% of the maximum sorption capacity at 15 min. The sorption isotherms of BPA were fitted well by Freundlich and linear sorption models. The sorption coefficients normalized by organic carbon contents were in the range of 665-878 at 10-30 degrees C. MLSS had significant effect on the sorption behavior. With the increase in MLSS, the sorption removal efficiency increased. Through the determination of thermodynamics parameters of BPA, it can be concluded that the sorption of BPA on the anaerobic sludge was mainly a physical process and partitioning between the water and sludge phases played a dominant role. Desorption tests showed that BPA sorption on anaerobic sludge was partially reversible.

[Experimental study of low dose irradiation for treatment of immuno-mediated aplastic anemia in mice].

As the lymphocytes of immuno-mediated aplastic anemia (IMAA) are in active state, and the hematopoietic stem cells are in silence, this study was aimed to design a new strategy to treat IMAA. To utilize the difference of radiosensitivity between active lymphocytes and silent hematopoietic stem cells, the animals suffered from IMAA were treated with a single low dose of irradiation, killing the active lymphocytes to release its suppression to hematopoietic stem cells without injuring the hematopoietic stem cells. Therefore, the hematopoiesis can be restored. Experiments were completed in IMAA mouse model. At day 4 after making IMAA, the model mice were giren total body irradiation of 150 cGy, the non-treated model mice and normal mice irradiated with 150 cGy were used as control. The survive time and survive rate of mice, blood picture, the account of nucleated cell of bone marrow, and pathological changes of bone marrow and lymphoid tissues of each group mice were observed. The results were as follows: (1) Survive rate of IMAA mice in non-treated group was 12.5%, the average survive time was 27.4 +/- 13.4 days. 100% of IMAA mice in irradiation-treated group survived over 60 days. The mice of irradiation control group all survived. (2) The account of WBC of IMAA mice in non-treated group dramatically decreased until to die, and in the irradiation-treated group it was gradually increased since the 10th day after treatment and close to normal level at the 28th day. (3) The RBC hematocrit of IMAA mice in non-treated group progressively decreased at day 14, and IMAA mice of irradiation-treated group gradually recovered closely to normal level after slightly fall at day 14, similar to the mice of irradiation control group. (4) The account of nucleated cells of bone marrow in non-treated IMAA mice dramatically decreased, and in the IMAA mice of the irradiation-treated group it was rapidly increased following transient fall, and restored to normal. (5) Pathological observations showed that the bone marrow and spleen of non-treated IMAA mice demonstrated typical aplastic anemia pattern, including bone marrow failure, marked splenatrophy, but the bone marrow and lymphoid tissues in the IMAA mice of irradiation-treated group were recovered to normal at day 28 after treatment. It is concluded that the low dose of irradiation displayed a significant therapeutic effect to IMAA mice, their hematopoisis could be completely restored to normal. The mechanism of therapeutic effect may contribute to low dose of irradiation killing the immunocompetent lymphocytes, therefore, suppressing hematopoiesis. The experiment results not only set up a new strategy for IMAA treatment, but also provided a clue to study the mechanism of IMAA.

[Toxicity reduction of synthetic municipal wastewater treated by anaerobic, anoxic or aerobic reactors].

Bioassay with Photobacterium phosphoreum was used to evaluate the toxicity of municipal wastewater. The toxicity reduction capabilities of three biological treatment processes (anaerobic reactor, anoxic reactor and aerobic reactor) treating the synthetic municipal wastewater were studied. Seven compounds including toluene, p-xylene, o-xylene, indole, pyridine, cyclohexanone and benzenepropanoic acid were selected to represent the main organic toxicants in the municipal wastewater. The GC-MS analysis was conducted to study the transformation of organic compounds in the effluents. The results show that in anaerobic reactor, the toxicity of effluent was higher than influent at short HRTs (no more than 10 h). The toxicity reduction capability of the anoxic reactor was in the medium level among the three reactors. The aerobic reactor had the strongest toxicity reduction capability.

[Activation of auto-mesenchymal stem cells of skeletal muscle by bone morphogenetic protein for rescuing bone marrow failure].

OBJECTIVE: To explore the effect of bone morphogenetic protein (BMP) to activate mesenchymal stem cells of skeletal muscle for rescuing bone marrow failure. METHODS: The study was performed on lethal rat acute aplastic anemia model induced by combined 5-fluorouracil (5-FU) and busulfan. The rh-BMP-2 was implanted into the thigh muscle of the rats at 3 days before aplastic anemia was induced. In the control group the rats were implanted with agar into the thigh muscle. The blood picture, pathologic changes and the mortality in two groups were observed. At the same time, rh-BMP-2 were implanted into the thigh muscle of normal Kun-min mice for dynamic control observation of the implantation local morphological changes, colony forming units-spleen (CFU-S) and stem cell growth factor (SCF) expression of the stroma cells of ectopic ossicles induced by BMP. RESULTS: At 7 days after BMP implantation in the mice the mesenchymal cells around BMP in muscle proliferated, and appeared in bone marrow to form an ectopic ossicles. The SCF expression of stroma cells in ectopic ossicles were higher than that of self-bone marrow. 56.3% of BMP-treated aplastic rats were survived over 3 months and its hematopoiesis was completely reconstituted and the histo-morphological picture of the spleen and bone marrow were recovered to normal. But in the control group only one of 23 rats was survived, the remainder died of hematopoietic failure. CONCLUSIONS: BMP-implantation into the skeletal muscle could rescue the bone marrow hematopoietic failure. The mechanism might be related to the BMP activated auto-mesenchymal cells of skeletal muscles to direct hematopoietic cell differentiation. In our hands it might create a new pathway for utilization of auto-muscle derived mesenchymal cells to reconstitute hematopoiesis.

[Study of Low Dose Irradiation Enhancing the Effect of Chemotherapeutic Agents against Leukemia and Its Mechanism]

For effectively enhancing the anti-leukemia effect of chemotherapeutic agents, and meanwhile decreasing the side effect of these agents, the study has been made to explore the synergistic effect of low dose irradiation (LDI) combined with Ara-C on murine leukemia and its mechanism. Firstly, an optimal scheme of low dose total body irradiation combined with Ara-C was established in L615 leukemia (T lymphocytic leukemia) mouse model. The machanism of the enhancing effect was explored by patho-morphological observation, examination of residual leukemia cells, the expression of GM-CSF on the surface of marrow stromal cells and in the bone marrow cultural supernatants. The results showed that the optimal scheme was 300 cGy irradiation at 4 days after inoculation of leukemic cells followed by Ara-C 30 mg/kg x 3 days in an interval of 1, 2 or 3 days after irradiation. The mean survival time of the L615 leukemia mice in LDI + Ara-C combined treatment groups was longer than that of control groups. The percentage of long-term survival mice (> 30 days) was the highest (58% - 72%), too. 17% of the mice were be cured. The numbers of blood leukocytes and marrow nucleated cells were transiently decreased in combined treatment group, and then recovered rapidly. Slight myelosuppression and marrow sinus dilation and congestion were seen after 300 cGy irradiation. The expression of GM-CSF either on the stromal cells or in marrow cultural supernatant after irradiation increased strikingly (P < 0.05). Therefore, LDI combined with Ara-C possesses synergistic effect. The mechanism is possibly related to three facts: LDI could increase the permeability of bone marrow sinus; LDI could promote marrow stromal cells to produce some cytokines (such as GM-CSF, etc.) which drive leukemia cells into cell cycle to make the cells more sensitive to chemotherapeutic agents; and LDI could augment Ara-C-induced cytotoxicity through the mechanism of apoptosis.