The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0（NH2-PAMAM G2.0）-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of...The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0（NH2-PAMAM G2.0）-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of Au（Ⅲ） to NH2-PAMAM G2.0 were studied by both UV-visible spectrometry and fluorospectrometry. The NH2-PAMAM G2.0-Au nanocomposites, with a type of structure in which one Au nanoparticle is surrounded by several NH2-PAMAM G2.0 dendrimers, emit strong bluish violet fluorescence, and are uniform, water soluble and biocompatible as well as very stable in frozen conditions. The size of gold nanoparticles in the nanocomposites is about 2.5 nm and decreases with the increase of NH2-PAMAM G2.0 concentration. The NH2-PAMAM G2.0 plays an important role in acting as host or micro-reactor for Au（Ⅲ） before Au（Ⅲ） reduction and acting as dispersant and stabilizer for gold nanoparticles after Au（Ⅲ） reduction. Preliminary experiments of cells staining to human embryonic lung fibroblast cell lines show that the NH2-PAMAM G2.0-Au nanocomposites can be used as optical imaging markers for bioanalyses and medical diagnoses.展开更多
We developed a special methylene blue solution for staining of cervix shedding cells based on catalytic oxidizing chromogenic reaction, which shows a potential for cervical cancer cytology screening. We screened a tot...We developed a special methylene blue solution for staining of cervix shedding cells based on catalytic oxidizing chromogenic reaction, which shows a potential for cervical cancer cytology screening. We screened a total of 1 922 women for cervical cancer with the special methylene blue staining method and a conventional Pap smear method using cervix shedding cells. Then, the patients with positive indicators of the Pap smear or this special solution staining method were examined by the electron colposcopy and histopathological examination. Staining of cervical exfoliated cells with this reactive oxygen-based special solution showed that the number of positive cases was 140(7.28%). Among them, 21 cases showed dark green(1.09%), and 119 cases showed purple black(6.19%). The results of the Pap smear method showed that the number of positive cases was 123(6.40%), of which ASCUS was 105(5.46%), ASC-H was 5(0.26%), and LSIL was 9(0.47%), and HSIL was 4(0.21%). For cervical exfoliated cell special staining solution for screening cervical intraepithelial neoplasia(CIN-Ⅱ, CIN-Ⅲ) and cervical cancer, sensitivity was 83.33%, specificity was 65.52%, accuracy was 74.29%, missed diagnosis rate was 13.33%, positive coincidence rate was 51.43%, and the negative coincidence rate is 86.67%. Our results proved the value of this method for early screening of cervical cancer through clinical practice in China.展开更多
Purpose:.To compare the merits and limitations of hematoxylin-eosin(HE) and methyl violet staining for displaying ghost cells from vitreous or aqueous humor.Methods:.A specimen containing ghost cells was adjusted to f...Purpose:.To compare the merits and limitations of hematoxylin-eosin(HE) and methyl violet staining for displaying ghost cells from vitreous or aqueous humor.Methods:.A specimen containing ghost cells was adjusted to five different concentrations:(12×104,.10×104,.8×104, 6×104and 4×104cells / ml) and subjected to smearing and methyl violet and HE staining..The staining results were observed by light microscopy.Results: The ghost cells were readily observed at a cell density of > 8×104cells / ml with methyl violet staining,.but only a few cells were occasionally seen at lower cell densities..In contrast,.ghost cells were seen at all cell densities with HE staining.Conclusion: Methyl violet staining is more rapid and simpler for the identification of ghost cells, but its staining color more readily fades, the slides cannot be stored, and it is only effective at a cell density of > 8 ×104cells / ml. In contrast,.HE staining is more time-consuming but it can display cell morphology and distinguish cell components more explicitly and slides can be permanently stored. HE staining has advantages over methyl violet staining in detecting the ghost cells when the concentration is < 8×104cells / ml.展开更多
[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA...[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA from six kinds of mycoplasma that commonly contaminated cells. Then the mycoplasma contamination of 25 cell samples was defected by PCR and DNA fluorescence staining. EResultl When these cell samples were detected by DNA fluorescence staining, the positive rate and probable positive rate were respectively 24% and 16%. And when they were detected by PCR, the positive rate was 36%. [ Condusion] The PCR method is more sensitive and specific than the DNA fluorescence staining, and combining these two methods is the optimal way to detect mycoplasma contamination in cell cultures.展开更多
Interleukin 17(IL-17)and its main producer,T cell receptorγδcells,have neurotoxic effects in the pathogenesis of intracerebral hemorrhage(ICH),aggravating brain injuries.To investigate the correlation between IL-17 ...Interleukin 17(IL-17)and its main producer,T cell receptorγδcells,have neurotoxic effects in the pathogenesis of intracerebral hemorrhage(ICH),aggravating brain injuries.To investigate the correlation between IL-17 and ICH,we dynamically screened serum IL-17 concentrations using enzyme-linked immunosorbent assay and explored the clinical values of IL-17 in ICH patients.There was a significant negative correlation between serum IL-17 level and neurological recovery status in ICH patients(r=–0.498,P<0.01).To study the neurotoxic role of IL-17,C57 BL/6 mice were used to establish an ICH model by injecting autologous blood into the caudate nucleus.Subsequently,the mice were treated with mouse neural stem cells(NSCs)and/or IL-17 neutralizing antibody for 72 hours.Flow cytometry,brain water content detection,Nissl staining,and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling results indicated that NSC transplantation significantly reduced IL-17 expression in peri-hematoma tissue,but there was no difference in T cell receptorγδcells.Compared with the ICH group,there were fewer apoptotic bodies and more Nissl bodies in the ICH+NSC group and the ICH+NSC+IL-17 group.To investigate the potential effect of IL-17 on directional differentiation of NSCs,we cultured mouse NSCs(NE-4 C)alone or co-cultured them with T cell receptorγδcells,which were isolated from mouse peripheral blood mononuclear cells,for 7 days.The results of western blot assays revealed that IL-17 secreted by T cell receptorγδcells reduced the differentiation of NSCs into astrocytes and neurons,while IL-17 neutralization relieved the inhibition of directional differentiation into astrocytes rather than neurons.In conclusion,serum IL-17 levels were elevated in the early stage of ICH and were negatively correlated with outcome in ICH patients.Animal experiments and cytological investigations therefore demonstrated that IL-17 probably has neurotoxic roles in ICH because of its inhibitory effects on the directional differentiation of NSCs.The application of IL-17 neutralizing antibody may promote the directional differentiation of NSCs into astrocytes.This study was approved by the Clinical Research Ethics Committee of Anhui Medical University of China(For human study:Approval No.20170135)in December 2016.All animal handling and experimentation were reviewed and approved by the Institutional Animal Care and Use Committee of Anhui Medical University(approval No.20180248)in December 2017.展开更多
文摘The solution chemical and optical characteristics of formation of amine-terminated polyamidoamine dendrimer G2.0（NH2-PAMAM G2.0）-Au nanocomposites in the aqueous solution of NH2-PAMAM G2.0 at various mole ratios of Au（Ⅲ） to NH2-PAMAM G2.0 were studied by both UV-visible spectrometry and fluorospectrometry. The NH2-PAMAM G2.0-Au nanocomposites, with a type of structure in which one Au nanoparticle is surrounded by several NH2-PAMAM G2.0 dendrimers, emit strong bluish violet fluorescence, and are uniform, water soluble and biocompatible as well as very stable in frozen conditions. The size of gold nanoparticles in the nanocomposites is about 2.5 nm and decreases with the increase of NH2-PAMAM G2.0 concentration. The NH2-PAMAM G2.0 plays an important role in acting as host or micro-reactor for Au（Ⅲ） before Au（Ⅲ） reduction and acting as dispersant and stabilizer for gold nanoparticles after Au（Ⅲ） reduction. Preliminary experiments of cells staining to human embryonic lung fibroblast cell lines show that the NH2-PAMAM G2.0-Au nanocomposites can be used as optical imaging markers for bioanalyses and medical diagnoses.
基金Supported by the Joint Foundation from Health Commission of Hubei Province(WJ2019H007)
文摘We developed a special methylene blue solution for staining of cervix shedding cells based on catalytic oxidizing chromogenic reaction, which shows a potential for cervical cancer cytology screening. We screened a total of 1 922 women for cervical cancer with the special methylene blue staining method and a conventional Pap smear method using cervix shedding cells. Then, the patients with positive indicators of the Pap smear or this special solution staining method were examined by the electron colposcopy and histopathological examination. Staining of cervical exfoliated cells with this reactive oxygen-based special solution showed that the number of positive cases was 140(7.28%). Among them, 21 cases showed dark green(1.09%), and 119 cases showed purple black(6.19%). The results of the Pap smear method showed that the number of positive cases was 123(6.40%), of which ASCUS was 105(5.46%), ASC-H was 5(0.26%), and LSIL was 9(0.47%), and HSIL was 4(0.21%). For cervical exfoliated cell special staining solution for screening cervical intraepithelial neoplasia(CIN-Ⅱ, CIN-Ⅲ) and cervical cancer, sensitivity was 83.33%, specificity was 65.52%, accuracy was 74.29%, missed diagnosis rate was 13.33%, positive coincidence rate was 51.43%, and the negative coincidence rate is 86.67%. Our results proved the value of this method for early screening of cervical cancer through clinical practice in China.
文摘Purpose:.To compare the merits and limitations of hematoxylin-eosin(HE) and methyl violet staining for displaying ghost cells from vitreous or aqueous humor.Methods:.A specimen containing ghost cells was adjusted to five different concentrations:(12×104,.10×104,.8×104, 6×104and 4×104cells / ml) and subjected to smearing and methyl violet and HE staining..The staining results were observed by light microscopy.Results: The ghost cells were readily observed at a cell density of > 8×104cells / ml with methyl violet staining,.but only a few cells were occasionally seen at lower cell densities..In contrast,.ghost cells were seen at all cell densities with HE staining.Conclusion: Methyl violet staining is more rapid and simpler for the identification of ghost cells, but its staining color more readily fades, the slides cannot be stored, and it is only effective at a cell density of > 8 ×104cells / ml. In contrast,.HE staining is more time-consuming but it can display cell morphology and distinguish cell components more explicitly and slides can be permanently stored. HE staining has advantages over methyl violet staining in detecting the ghost cells when the concentration is < 8×104cells / ml.
基金Supported by Key Project of Anhui Province Natural Science Foundation(KJ2008A085)Key Sci-tech Research Project of Anhui Province(08010302179)2008 NSFC General Project of China ( 30872253)~~
文摘[ Objective ] To develop a rapid efficient method for detecting mycoplasma contamination in cell cultures. [ Method] A pair of primers was designed according to two highly conserved nucleotide sequences of the 16S RNA from six kinds of mycoplasma that commonly contaminated cells. Then the mycoplasma contamination of 25 cell samples was defected by PCR and DNA fluorescence staining. EResultl When these cell samples were detected by DNA fluorescence staining, the positive rate and probable positive rate were respectively 24% and 16%. And when they were detected by PCR, the positive rate was 36%. [ Condusion] The PCR method is more sensitive and specific than the DNA fluorescence staining, and combining these two methods is the optimal way to detect mycoplasma contamination in cell cultures.
基金supported by the Natural Science Foundation of Anhui Province of China,No.1708085MH211(to HWC)the College Top-notch Talent Foundation of Anhui Province of China,No.KJ2018A0207(to HWC)
文摘Interleukin 17(IL-17)and its main producer,T cell receptorγδcells,have neurotoxic effects in the pathogenesis of intracerebral hemorrhage(ICH),aggravating brain injuries.To investigate the correlation between IL-17 and ICH,we dynamically screened serum IL-17 concentrations using enzyme-linked immunosorbent assay and explored the clinical values of IL-17 in ICH patients.There was a significant negative correlation between serum IL-17 level and neurological recovery status in ICH patients(r=–0.498,P<0.01).To study the neurotoxic role of IL-17,C57 BL/6 mice were used to establish an ICH model by injecting autologous blood into the caudate nucleus.Subsequently,the mice were treated with mouse neural stem cells(NSCs)and/or IL-17 neutralizing antibody for 72 hours.Flow cytometry,brain water content detection,Nissl staining,and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling results indicated that NSC transplantation significantly reduced IL-17 expression in peri-hematoma tissue,but there was no difference in T cell receptorγδcells.Compared with the ICH group,there were fewer apoptotic bodies and more Nissl bodies in the ICH+NSC group and the ICH+NSC+IL-17 group.To investigate the potential effect of IL-17 on directional differentiation of NSCs,we cultured mouse NSCs(NE-4 C)alone or co-cultured them with T cell receptorγδcells,which were isolated from mouse peripheral blood mononuclear cells,for 7 days.The results of western blot assays revealed that IL-17 secreted by T cell receptorγδcells reduced the differentiation of NSCs into astrocytes and neurons,while IL-17 neutralization relieved the inhibition of directional differentiation into astrocytes rather than neurons.In conclusion,serum IL-17 levels were elevated in the early stage of ICH and were negatively correlated with outcome in ICH patients.Animal experiments and cytological investigations therefore demonstrated that IL-17 probably has neurotoxic roles in ICH because of its inhibitory effects on the directional differentiation of NSCs.The application of IL-17 neutralizing antibody may promote the directional differentiation of NSCs into astrocytes.This study was approved by the Clinical Research Ethics Committee of Anhui Medical University of China(For human study:Approval No.20170135)in December 2016.All animal handling and experimentation were reviewed and approved by the Institutional Animal Care and Use Committee of Anhui Medical University(approval No.20180248)in December 2017.
