Establishment of a Tm-shi Method for Detection of Cat-Derived Hookworms

Yeqi Fu,Yunqiu Liu,Asmaa M.I. Abuzeid,Yue Huang,Xue Zhou,Long He,Qi Zhao,Xiu Li,Jumei Liu,Rongkun Ran,Guoqing Li 대한기생충학ㆍ열대의학회 2019 The Korean Journal of Parasitology Vol.57 No.1

Melting temperature shift (Tm-shift) is a new detection method that analyze the melting curve on real-time PCR thermocycler using SYBR Green I fluorescent dye. To establish a Tm-shift method for the detection of Ancylostoma ceyl- anicum and A. tubaeforme in cats, specific primers, with GC tail of unequal length attached to their 5 ́ end, were designed based on 2 SNP loci (ITS101 and ITS296) of the internal transcribed spacer 1 (ITS1) sequences. The standard curve of Tm-shift was established using the standard plasmids of A. ceylanicum (AceP) and A. tubaeforme (AtuP). The Tm-shift method stability, sensitivity, and accuracy were tested with reference to the standard curve, and clinical fecal samples were also examined. The results demonstrated that the 2 sets of primers based on the 2 SNPs could accurately distin- guish between A. ceylanicum and A. tubaeforme. The coefficient of variation (CV) of Tm- values of AceP and AtuP was 0.07% and 0.06% in ITS101 and was 0.06% and 0.08% in ITS296, respectively. The minimum detectable DNA concen- tration was 5.22×10-6 and 5.28×10-6 ng/μl samples of AceP and AtuP, respectively. The accuracy of Tm-shift method reached 100% based on examination of 10 hookworm DNA samples with known species. In the clinical detection of hookworm in 69 stray cat fecal sample, the Tm-shift detection results were consistent with the microscopic examination and successfully differentiated between the 2-hookworm species. In conclusion, the developed method is a rapid, sensi- tive and accurate technique and can provide a promising tool for clinical detection and epidemiological investigation of cat-derived hookworms.

Enhancement mechanism of SO2 removal with calcium hydroxide in the presence of NO2

Jihui Gao,Guoqing Chen,Xiaolin Fu,Yijun Yin,Shaohua Wu,Yukun Qin 한국화학공학회 2012 Korean Journal of Chemical Engineering Vol.29 No.2

The enhancement mechanism of SO2 removal by the presence of NO2 under low temperature and humid conditions was studied in a fixed bed reactor system. The presence of NO2 in the flue gas can enhance SO2 removal. The interaction between SO2 and NO2 in gas phase could not explain the effect of NO2 on SO2 removal under lowtemperature and humid conditions. When Ca(NO3)2 and Ca(NO2)2 as additive were added on the surface of sorbent,the desulfurization activity of sorbent decreased. However, the sorbent pretreated by NO2 for a moment has higher SO2removal. The oxidization of SO32− to SO42− and the evolution of sorbent surface structure in the presence of NO2 can explain the enhancement of SO2 removal by the presence of NO2. HSO3− and SO3− reacted with NO2 to form sulfate,which can accelerate the hydrolysis of SO2. The reaction between NO2 and Ca(OH)2 can make the unreacted sorbet under the SO2 removal product exposed to the reactant gas.

Di-2-ethylhexyl phthalate induced oxidative damage involving FasL-associated apoptotic pathway in mouse spermatogenic GC-2spd cells

Xiao Tang,Kun Tong,Lishan Zhu,Guoqing Fu,Wei Chang,Ting Zhou,Zhibing Zhang,Ling Tong,Ling Zhang,Yuqin Shi,Ling Zhang,Yuqin Shi 대한독성 유전단백체 학회 2016 Molecular & cellular toxicology Vol.12 No.4

Di-2-ethylhexyl phthalate (DEHP) which belongs to phthalatic-acid esters, is a kind of harmful, global environmental pollutants. It is a known endocrine disrupting chemical and male reproductive toxicant. However, the mechanism by which DEHP exposure result in male reproductive toxicity is still unclear. To elucidate the productive toxicity mechanism of DEHP, we attemptted to investigate oxidative stress, apoptotic effects, mRNA and protein expression of apoptosis-associated genes including FasL, caspase-3, and caspase-8 in GC-2spd (mouse spermatogenic cells). The results showed that, with the increase of DEHP concentration, cell apoptosis rate increased; the activities of relation index of oxidative stress such as malondialdehyde (MDA), superoxide dismutase (SOD) and glutatione peroxidase (GSH-PX) changed significantly; the mRNA and protein expression levels of FasL, caspase-3, -8, altered obviously. These results suggested that DEHP could induce apoptosis of GC-2spd cells through oxidative stress and FasL-dependent pathway.

Evaluation of the Immunomodulatory and Anti-inflammatory Activities of Honey Bee Larva Powder

Kejuan Li,Shuang Sun,Masakatsu Kageyama,Long Xiao,Guoqing Xing,Ran Gao,Fengming You,Xi Fu,Zhenya Zhang 한국식품영양과학회 2020 Journal of medicinal food Vol.23 No.7

Honey bee larva powder (HLP) has traditionally been used as a daily supplement and tonic for health promotion with an uncertain scientific basis. In this study, B16-F10 tumor-bearing mice were established to evaluate the immunomodulatory activity of HLP. The proliferation and apoptosis assays were performed to evaluate the anti-inflammatory activity of honey bee larva extract (HLE) in RAW 264.7 macrophage. The in vivo experimental results demonstrated that the oral administration of freeze-dried HLP (4 and 6 g/kg) significantly enhanced the spleen index, the percentage of CD4+cells, and the ratio of CD4+ and CD8+ T lymphocytes (CD4+/CD8+) in the peripheral blood compared with those in the tumor control mice. The in vitro studies demonstrated the potent immunomodulatory activities of HLE through the induction of RAW 264.7 macrophage proliferation and the mitigation of doxorubicin (DOX)-induced toxicity. HLE also exhibits anti-inflammatory activity by decreasing the production of nitric oxide (NO) and the cytokine level of interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage. The present study provides important scientific evidence for the immunomodulatory and anti-inflammatory activities of HLP and HLE.

Effects of Acute Sacral Neuromodulation at Different Pulse Widths on Bladder Overactivity in Pigs

Huiling Cong,Limin Liao,Yiming Wang,Lingna Zhao,Zhaoxia Wang,Guang Fu,Guoqing Chen 대한배뇨장애요실금학회 2019 International Neurourology Journal Vol.23 No.2

Purpose: Sacral nerve stimulation has been used to treat overactive bladder. This study evaluated the effects of stimulation using different pulse widths on the inhibition of bladder overactivity by sacral nerve stimulation (SNM) in pigs. Methods: Implant-driven stimulators were used to stimulate the S3 spinal nerve in 7 pigs. Cystometry was performed by infusing normal saline (NS) or acetic acid (AA). SNM at pulse widths of 64 μsec to 624 μsec was conducted at the intensity threshold at which observable perianal and/or tail movement was induced. Multiple cystometrograms were performed to determine the effects of different pulse widths on the micturition reflex. Results: AA-induced bladder overactivity reduced the bladder capacity to 46.9%±7.1% of the NS control level (P<0.05). During AA infusion, SNM at 64 μsec, 204 μsec, and 624 μsec increased the bladder capacity to 126.1%±6.9%, 129.5%±7.3%, and 140.1%±7.6% of the AA control level (P<0.05). No significant differences were found among the results obtained using pulse widths of 64 μsec, 204 μsec, and 624 μsec (P>0.05). The actual intensity threshold varied from 0.7 to 8 V. The mean intensity threshold (T visual) for pulse widths of 64 μs, 204 μs, and 624 μs were 5.64±0.76 V, 3.11±0.48 V, and 2.52±0.49 V. T visual for pulse widths of 64 μsec was larger than the other two T visual for pulse widths of 204 μsec and 624 μsec (P<0.05). No significant differences were found among the T visual for pulse widths of 204 μsec and 624 μsec (P>0.05). Conclusions: This study indicated that different pulse widths could play a role in inhibiting bladder overactivity. It is not yet certain which pulse widths increased bladder capacity compared with AA levels, to minimize energy consumption and maintain patient comfort during stimulation, 204 μsec may be an appropriate pulse width for SNM.

Sequence Analysis of Mitochondrial Genome of Toxascaris leonina from a South China Tiger

Kangxin Li,Fang Yang,A. Y. Abdullahi,Meiran Song,Xianli Shi,Minwei Wang,Yeqi Fu,Weida Pan,Fang Shan,Wu Chen,Guoqing Li 대한기생충학열대의학회 2016 The Korean Journal of Parasitology Vol.54 No.6

Toxascaris leonina is a common parasitic nematode of wild mammals and has significant impacts on the protection of rare wild animals. To analyze population genetic characteristics of T. leonina from South China tiger, its mitochondrial (mt) genome was sequenced. Its complete circular mt genome was 14,277 bp in length, including 12 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 2 non-coding regions. The nucleotide composition was biased toward A and T. The most common start codon and stop codon were TTG and TAG, and 4 genes ended with an incomplete stop codon. There were 13 intergenic regions ranging 1 to 10 bp in size. Phylogenetically, T. leonina from a South China tiger was close to canine T. leonina. This study reports for the first time a complete mt genome sequence of T. leonina from the South China tiger, and provides a scientific basis for studying the genetic diversity of nematodes between different hosts.