[Structural Analysis of Salivary Bacterial and Fungal Microbiome in Oral Lichen Planus Patients].

Objective: To explore the differences of oral mycobiome and bacteriome between the healthy controls (H) and oral lichen planus (OLP) patients, and the co-occurrence patterns of the salivary mycobiome and bacteriome and the association with host immunity. Methods: Saliva samples were collected from clinical OLP patients (n=35) and healthy volunteers (n=18). Microbiome DNA was extracted for bacterial 16S rRNA genes sequencing and fungal internal transcribed spacer 2 (ITS2) sequencing. Bioinformatics analysis was performed on the data.The levels of IL-17 and IL-23, two pro-inflammatory cytokines, in the saliva were examined, and their correlation with the bacteria was analyzed. Results: There was no significant difference in the overall community structure of the mycobiome and the bacteriome between OLP patients and healthy controls. The abundance of Prevotellaand Solobacterium in the saliva bacteriome was significantly increased in the OLP group (P<0.05), and the relative abundance of Candidaand Aspergillusin the saliva mycobiome was also significantly increased (P<0.05). The co-occurrence pattern of the salivary mycobiome and bacteriome showed that the aforementioned difference was not related. However, the correlation between Aspergillusand bacteria was altered in the H group and the OLP group, and co-occurrence was reduced in the latter group. The level of IL-17 in the saliva was significantly increased in the OLP group. IL-17 and clinical scores were significantly correlated with the abundance of Porphyromonas. Conclusion: The increased abundance of Prevotella, Solobacterium, Candida, and Aspergillus was associated with the pathogenesis of OLP, and the changes of the microbiome co-occurrence relationship and host immunity may be involved in the pathogenesis of OLP.

Anti-H7N9 avian influenza A virus activity of interferon in pseudostratified human airway epithelium cell cultures.

BACKGROUND: Since H7N9 influenza A virus (H7N9) was first reported in 2013, five waves of outbreaks have occurred, posing a huge threat to human health. In preparation for a potential H7N9 epidemic, it is essential to evaluate the efficacy of anti-H7N9 drugs with an appropriate model. METHODS: Well-differentiated pseudostratified human airway epithelium (HAE) cells were grown at the air-liquid interface, and the H7N9 cell tropism and cytopathic effect were detected by immunostaining and hematoxylin-eosin (HE) staining. The H7N9 replication kinetics and anti-H7N9 effect of recombinant human α2b (rhIFN-α2b) and rhIFN-λ1 were compared with different cell lines. The H7N9 viral load and interferon-stimulated gene (ISG) expression were quantified by real-time PCR assays. RESULTS: H7N9 could infect both ciliated and non-ciliated cells within the three-dimensional (3D) HAE cell culture, which reduced the number of cilia and damaged the airways. The H7N9 replication kinetics differed between traditional cells and 3D HAE cells. Interferon had antiviral activity against H7N9 and alleviated epithelial cell lesions; the antiviral activity of rhIFN-α2b was slightly better than that of rhIFN-λ1. In normal cells, rhIFN-α2b induced a greater amount of ISG expression (MX1, OAS1, IFITM3, and ISG15) compared with rhIFN-λ1, but in 3D HAE cells, this trend was reversed. CONCLUSIONS: Both rhIFN-α2b and rhIFN-λ1 had antiviral activity against H7N9, and this protection was related to the induction of ISGs. The 3D cell culture model is suitable for evaluating interferon antiviral activity because it can demonstrate realistic in vivo-like effects.

The effects of aerobic exercise on the structure and function of DMN-related brain regions: a systematic review.

Physical activity may play a role in both the prevention and slowing of brain volume loss and may be beneficial in terms of improving the functional connectivity of brain regions. But much less is known about the potential benefit of aerobic exercise for the structure and function of the default mode network (DMN) brain regions. This systematic review examines the effects of aerobic exercise on the structure and function of DMN brain regions in human adulthood. Seven electronic databases were searched for prospective controlled studies published up to April 2015. The quality of the selected studies was evaluated with the Cochrane Collaboration's tool for assessing the risk of bias. RevMan 5.3 software was applied for data analysis. Finally, 14 studies with 631 participants were identified. Meta-analysis revealed that aerobic exercise could significantly increase right hippocampal volume (SMD = 0.26, 95% CI 0.01-0.51, p = 0.04, I2 = 7%, 4 studies), and trends of similar effects were observed in the total (SMD = 0.12, 95% CI -0.17 to 0.41, p = 0.43, I2 = 0%, 5 studies), left (SMD = 0.12, 95% CI -0.13 to 0.37, p = 0.33, I2 = 14%, 4 studies), left anterior (SMD = 0.12, 95% CI -0.16 to 0.40, p = 0.41, I2 = 74%, 2 studies) and right anterior (SMD = 0.10, 95% CI -0.17 to 0.38, p = 0.46, I2 = 76%, 4 studies) hippocampal volumes compared to the no-exercise interventions. A few studies reported that relative to no-exercise interventions, aerobic exercise could significantly decrease the atrophy of the medial temporal lobe, slow the anterior cingulate cortex (ACC) volume loss, increase functional connectivity within the hippocampus and improve signal activation in the cingulate gyrus and ACC. The current review suggests that aerobic exercise may have positive effects on the right hippocampus and potentially beneficial effects on the overall and other parts of the hippocampus, the cingulate cortex and the medial temporal areas of the DMN. Moreover, aerobic exercise may increase functional connectivity or activation in the hippocampus, cingulate cortex and parahippocampal gyrus regions of the DMN. However, considering the quantity and limitations of the included studies, the conclusion could not be drawn so far. Additional randomized controlled trials (RCTs) with rigorous designs and longer intervention periods are needed in the future.

Synthesis and cytotoxic activity of two steroids: icogenin aglycone analogs.

During the process of icogenin analog research, we obtained two cytotoxic steroids: compound 4 and compound 6 casually. Their in vitro antitumor activities were tested by the standard MTT assay. The results disclosed that compound 4 (IC50 = 3.65-6.90 µM) showed potential antitumor activities against HELA, KB cell lines and compound 6 (IC50 = 2.40-9.05 µM) showed potential antitumor activities against HELA, BGC-823, KB, A549, HCT-8 cell lines.

[Relationship between Electroencephalogram and Perceived Stress in Essential Hypertension Patients].

OBJECTIVE: To explore the relationship between electroencephalogram and perceived stress in Chinese essential hypertension (EH) patients. METHODS: Totally 60 EH patients were enrolled as the case group,and 60 gender-and age-matched healthy subjects as the control group. Graded mental arithmetic was used as a psychological stressor. The physiological values of electroencephalogram (Theta,Alpha,sensorimotor rhythm,and Beta) were recorded at baseline and in mission period and recovery period. Subjects were instructed to complete the Perceived Stress Scale. RESULTS: The electroencephalogram Theta value was significantly different between the case group and the control group (t=4.01,P=0.048). In the case group,the fluctuating Theta value was correlated with the control dimension in the Perceived Stress Scale (r=-0.26,P=0.044). Also in the case group,with the score of "sense of control" as the dependent variable and the fluctuating value of Theta after stress as the independent variables,regression analysis was performed. The opt-in standard was≤0.05 and the culling standard was≥0.01. The Results shows that the fluctuating value of Theta after stress in the case group met the requirements to enter the equation and could explain 5.2% of "sense of control" dimension. CONCLUSION: The Theta values associated with the sleeping and creativity decrease under stressful situations in EH patients following the increaed sense of pressure due to failure in controlling the pressure events.

MicroRNA-190 regulates hypoxic pulmonary vasoconstriction by targeting a voltage-gated K⁺ channel in arterial smooth muscle cells.

Pulmonary arterial hypertension (PAH) is associated with sustained vasoconstriction, profound structural remodeling of vasculatures and alterations in Ca(2+) homeostasis in arterial smooth muscle cells (SMCs), while the underlying mechanisms are still elusive. By regulating the expression of proteins, microRNAs (miRNAs) are known to play an important role in cell fates including differentiation, apoptosis and proliferation, and may be involved in the development of PAH. Based on our previous study, hypoxia produced a significant increase of the miR-190 level in the pulmonary artery (PA), here, we used synthetic miR-190 to mimic the increase in hypoxic conditions and showed evidence for the effects of miR-190 on pulmonary arterial vasoconstriction and Ca(2+) influx in arterial SMCs. Synthetic miR-190 remarkably enhanced the vasoconstriction responses to phenylephrine (PE) and KCl. The voltage-gated K(+) channel subfamily member, Kcnq5, mRNA was shown to be a target for miR-190. Meanwhile, miR-190 antisense oligos can partially reverse the effects of miR-190 on PASMCs and PAs. Therefore, these results suggest that miR-190 appears to be a positive regulator of Ca(2+) influx, and plays an important role in hypoxic pulmonary vascular constriction.

Relationship between cardiovascular responses and coping styles among recruits under the stress.

OBJECTIVE: To explore the relationship between the cardiovascular responses and coping styles among recruits in psychological stress. METHODS: Using random cluster sampling method,we chose 406 army recruits in Xinjiang from October 2013 to November 2013,and the questionnaire survey combined with laboratory test was applied. The questionnaire used in this study was the simplified coping style questionnaire. Systolic blood pressure(SPB),diastolic blood pressure(DPB),and heart rate(HR)were recorded. The relationship between cardiovascular responses and coping methods was analyzed using descriptive statistics,correlations,and regression analysis. RESULTS: There were statistically significant differences in changes of the indicators of cardiovascular responses(P=0.000). The positive coping styles were significantly associated with age(P=0.008)and father's education degree(P=0.010). The negative coping styles were associated with the mother's educational degree(P=0.046). The positive coping styles were associated with the SBP in recall task period(P=0.039)and with the HR in calculating task period(P=0.025). The negative coping styles were significantly associated with the SBP difference between the baseline and calculating task period(P=0.034). Stepwise regression analysis showed that age and father's education degree(P=0.008,P=0.041)had a positive effect on the positive coping styles,and the SPB difference between the baseline and calculating task period had a negative effect on the positive coping styles(P=0.034). CONCLUSIONS: Age and father's education degree can affect the positive coping style of the recruits. Increased psychological stress has a negative impact on the negative coping styles of the recruits.

[Comparing hydroxyapatite coated versus non hydroxyapatite coated femoral stems in primary total hip arthroplasty: a meta analysis of randomized controlled trial].

OBJECTIVE: To evaluate the difference of clinical outcomes and radiological outcomes through meta-analysis on the total hip arthroplasty (THA) between hydroxyapatite(HA) coating and non-HA coating femoral stems. METHODS: We searched the MEDLINE, Embase, Cochrane library and CBM for published randomized controlled trial (RCT) comparing HA coating and non-HA coating femoral stems in primary THA clinical outcomes with Harris hip score and incidence postoperative thigh pain, radiological outcomes with presence of endosteal condensation and radioactive line on the prothesis, heterotopic ossification. Data analysis were performed using RevMan 5.0(the Cochrane Collaboration). RESULTS: Ten studies and 917 hips into our analysis, with 464 hips in HA groups and 453 hips in non-HA groups. The combined results of the meta-analysis indicated there was no statistical differences between the two groups on postoperative Harris hip score(WMD = 3.04, 95%CI:-4.47-10.54, P = 0.43) , there was statistical difference on incidence postoperative thigh pain (RR = 0.56, 95%CI:0.33-0.94, P = 0.03) . There were no significant differences between the two groups on presence of endosteal condensation (RR = 1.01, 95%CI:0.91-1.11, P = 0.91), presence of radioactive line (RR = 0.99, 95%CI:0.88-1.11, P = 0.83) and incidence of heterotopic ossification (RR = 0.97, 95%CI:0.77-1.21, P = 0.77). CONCLUSIONS: There are no clinical and radiological benefits in the use of HA coating femoral stems in Primary THA, there is not enough evidence prove the HA can reduce the incidence postoperative thigh pain.

[Effect of different host plants on the volatile components in Taxillus sutchuenensis].

OBJECTIVE: To study the effect of different host plants on the volatile somponents in Taxillus sutchuenensis. METHODS: Volatile oils of T. sutchuenensis from two different hosts (Magnolia denudata and Camellia oleifera) were obtained by steam distillation. The chemical constituents and relative contents of the volatile oils were analyzed by GC-MS. RESULTS: There were some similarities of volatile constituents between T. sutchuenensis and the corresponding host plants while the volatile components of T. sutchuenensis isolated from different host plants were not the same. CONCLUSION: Host plants have effect on volatile constituents of T. sutchuenensis apparently.

[Effects of ERα gene overexpression on bone mineral density and calcium and phosphorus metabolism inovariectomized osteoporosis mice].

Objective: To investigate the effects of estrogen receptor α (ERα) gene overexpression on bone metabolism and calcium and phosphorus metabolism in ovariectomized osteoporosis mice, and to provide experimental basis for targeted gene therapy of osteoporosis. Methods: Thirty SPF female mice were randomly divided into sham operation group, model group and ERα overexpression group with 10 mice in each group. After the model was established, the ERα overexpression group was transfected with recombinant adenovirus vector carrying mouse ERα gene by intraspinal injection. The model group was transfected with empty virus, and the sham operation group was not treated. The expression of ERα gene in bone tissue of mice was detected by quantitative Real-time PCR (qRT-PCR). Bone mineral density (BMD) of mouse femur was measured after modeling. Trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular segregation (Tb.Sp), bone volume fraction (BV/TV) and biomechanical strength of femur were measured by micro-CT scanning. Serum levels of calcium (Ca), phosphorus (P), osteocalcin (BGP) and alkaline phosphatase (ALP) were measured by automatic biochemical analyzer. The expressions of tissue inhibitor of metalloproteinases 1 (TIMP-1) and monocyte chemotactic protein 1 (MCP-1) in bone homogenate were detected by Immunohistochemistry. Results: Compared with sham operation group, the expression level of ERα gene in bone tissue of model group was decreased significantly, the levels of BMD, BV/TV, Tb. Th, maximum load, rigidity coefficient, Ca and P were decreased, while the levels of Tb. Sp, BGP and ALP were increased significantly (P＜0.05). Compared with the sham operation group, the expression level of TIMP-1 protein in the bone tissue of the model group was significantly decreased, while that of MCP-1 protein was increased, while that of the ERα overexpression group was increased while that of MCP-1 was decreased (P＜0.05).The levels of ERα gene expression, BMD, BV/TV, TB. Th, maximum load, rigidity coefficient, Ca and P in the ERα overexpression group were significantly higher than those in the model group, while Tb. Sp, BGP and ALP were significantly lower (P＜0.05). Compared with the sham operation group, mean optical density of TIMP-1 in the bone tissue of the model group was significantly decreased, while that of MCP-1 was significantly increased, and that of the ERα overexpression group was significantly increased while that of MCP-1 was significantly decreased (P＜0.05). Conclusion: ERα gene overexpression can improve osteoporosis by regulating bone mineral density, bone parameters, bone metabolism, calcium and phosphorus metabolic indicators and the expression levels of TIMP-1 and MCP-1 in tissues.

Endometrial Preparation for Frozen-Thawed Embryo Transfer With or Without Pretreatment With GnRH Agonist: A Randomized Controlled Trial at Two Centers.

Objective: This prospective randomized controlled trial compared the reproductive outcomes of frozen embryo transfer (FET) with hormone replacement treatment (HRT) with or without gonadotropin-releasing hormone agonist (GnRHa) pretreatment. Methods: A total of 133 patients scheduled for HRT-FET mainly because of tubal and/or male factors who received two high-quality cleavage-stage embryos were enrolled at two participating centers. The GnRHa group (n = 65) received GnRHa pretreatment, while the control group (n = 68) did not. Analysis was based on the intention-to-treat (ITT) principle. Results: Among the 133 participants, 130 (97.7%) underwent embryo transfer and 127 (95.5%) completed the protocol. The clinical pregnancy rate according to ITT did not differ between the GnRHa and control groups [39/65 (60.0%) vs. 41/68 (60.3%), p = 0.887]. The implantation rate (47.6% vs. 45.3%, p = 0.713), early pregnancy loss rate (5.1% vs. 19.5%, p = 0.09), and live birth rate (49.2% vs. 50.0%, p = 0.920) were also comparable between groups. Conclusion: Pretreatment with GnRHa does not improve the reproductive outcomes for women receiving HRT-FET. Clinical Trial Registration: The study was registered with the Chinese Clinical Trial Registry (ChiCTR-IOR-17014170; http://www.chictr.org.cn).

Genome-wide analysis of the AINTEGUMENTA-like (AIL) transcription factor gene family in pumpkin (Cucurbita moschata Duch.) and CmoANT1.2 response in graft union healing.

AINTEGUMENTA-like (AIL) proteins are members of the APETALA 2/ETHYLENE RESPONSE FACTOR (AP2/ERF) domain family of transcription factors involved in plant growth, development, and abiotic stress responses. However, the biological functions of AIL members in pumpkin (Cucurbita moschata Duch.) remain unknown. In this study, we identified 12 AIL genes in the pumpkin genome encoding proteins predicted to be localized in the nucleus. Phylogenetic analysis showed that the AIL gene family could be classified into six major subfamilies, with each member encoding two AP2/ERF domains separated by a linker region. CmoAIL genes were expressed at varying levels in the examined tissues, and CmoANT genes showed different expression patterns under auxin (IAA), 1-naphthylphthalamic acid (NPA), and abscisic acid (ABA) treatments. Ectopic overexpression of CmoANT1.2 in Arabidopsis increased organ size and promoted growth of grafted plants by accelerating graft union formation. However, there was no significant difference at the graft junction for WT/WT and WT/ANT under IAA or NPA treatments. Taken together, the results of this study provide critical information about CmoAIL genes and their encoded proteins, and suggest future work should investigate the functions of CmoANT1.2 in the grafting process in pumpkin.

Sequence polymorphism and evolution of three cetacean MHC genes.

Sequence variability at three major histocompatibility complex (MHC) genes (DQB, DRA, and MHC-I) of cetaceans was investigated in order to get an overall understanding of cetacean MHC evolution. Little sequence variation was detected at the DRA locus, while extensive and considerable variability were found at the MHC-I and DQB loci. Phylogenetic reconstruction and sequence comparison revealed extensive sharing of identical MHC alleles among different species at the three MHC loci examined. Comparisons of phylogenetic trees for these MHC loci with the trees reconstructed only based on non-PBR sites revealed that allelic similarity/identity possibly reflected common ancestry and were not due to adaptive convergence. At the same time, trans-species evolution was also evidenced that the allelic diversity of the three MHC loci clearly pre-dated species divergence events according to the relaxed molecular clock. It may be the forces of balancing selection acting to maintain the high sequence variability and identical alleles in trans-specific manner at the MHC-I and DQB loci.

[Application of the interaction teaching mode integrated with virtual anatomy platform in teaching Meridian and Acupoints].

OBJECTIVE: To observe the effect of the interaction teaching mode integrated with Visible Body virtual anatomy platform in teaching Meridian and Acupoints. METHODS: A total of 60 students in the class of 2017 in the discipline of acupuncture-moxibustion and tuina, Xiangnan University were recruited and randomized into an observation group and a control group, 30 students in each one. In the control group, the traditional practical teaching mode was used. In the observation group, the interaction teaching mode integrated with virtual anatomy platform was adopted. The teaching duration was 10 class hours in both groups. After accomplishing the teaching schedule, the practical examination was conducted in the localization of commonly-used acupoints, very useful acupoints and the dangerous acupoints as well as acupuncture manipulation techniques. Moreover, the degree of satisfaction was investigated among the students in the two groups and the self-learning ability was evaluated in 3-month follow-up visit. RESULTS: In the observation group, the scores for the localization and acupuncture manipulation of commonly-used acupoints, very useful acupoints and the dangerous acupoints, as well as the degree of satisfaction of the 3 items, i.e. interesting, interaction and leaning-assistance were all higher than those in the control group (P<0.05). The degree of satisfaction in the acceptance and leaning-participation, as well as the scores of self-learning ability in 3-month follow-up visit were not different statistically between the two groups (P>0.05). CONCLUSION: The interaction teaching mode integrated with virtual anatomy platform improves the effect on teaching Meridian and Acupoints and achieves the high student satisfaction.

[Continuous axenic cultivation of Pneumocystis carinii isolated from the bronchoalveolar lavage fluid of infected rat].

OBJECTIVE: To establish axenic cultivation of Pneumocystis carinii (P.c). METHODS: The organisms of P.c were isolated from the bronchoalveolar lavage fluid (BALF) of the rats with Pneumocystis carinii pneumonia (PCP) and cultured in a medium which was based on IMDM(GIBCO) supplemented with S-adenosyl-L-methionine, putrescine, N-acetyl glucosamine, putrescine, L-cysteine and L-glutamine, and newborn calf serum. The organisms cultured in the system were identified by observing the morphology of cysts in smears stained with Gomori's methenamine silver nitrate stain (GMS). Ultrastructure of the cysts/trophozoites was examined by transmission electron microscopy. The sequences of mitochondrial large ribosomal DNA subunit of the cultured organisms were compared with the Pneumocystis carinii f.sp. ratti variant isolate (GenBank No U20173) and Pneumocystis carinii f.sp.hominis (GenBank No M58605). RESULTS: Five isolates of P. carinii received from BALF of 8 rats with PCP were cultured axenically and continuously in the system. The cultured organisms could be stored in frozen condition and used to reinitiate culture, and were amplified by 19-22 times within 72 h. The morphology, ultrastructure and gene sequencing of the cultured organisms confirmed that the isolated organisms were P. carinii. CONCLUSION: Five continuously and axenicly cultured isolates of P. carinii have been received.

[Study on hepatitis B virus intrauterine infection state and its correlation factors].

OBJECTIVE: To investigate the correlation factors of hepatitis B virus (HBV) intrauterine infection and the influence factors of HBV infection in peripheral blood mononuclear cells (PBMC) and placentas. METHODS: HBeAg and HBsAg in 151 pregnant women and their newborns were determined by enzyme linked immunoadsorbent assay (ELISA). HBV DNA in serum and PBMC of pregnant women and their newborns were determined by polymerase chain reaction. HBsAg in 151 placentas were detected by immunohistochemistry ABC. The correlation risk factors were analyzed by non-condition logistic regression model. RESULTS: HBV DNA positive in serum, HBV DNA positive in PBMC of pregnant women and HBsAg positive in placentas were the risk factors for HBV intrauterine infection. Their odds ratio (OR) and 95% confidence interval were 2.25 (1.08-4.72), 2.69 (1.26-5.73) and 4.63 (1.70-12.62), respectively. The influence factors of HBV infection in placenta included antepartum injection of hepatitis B immunoglobulin (HBIG) over thrice and HBV DNA positive in serum of pregnant women with OR 0.08 (0.01-0.69) and 4.24 (1.22-14.69). The risk factor for HBV DNA positive in PBMC of newborns was HBV DNA positive in PBMC of their mothers with OR 24.53 (7.92-76.01). CONCLUSIONS: HBV infection in placentas, HBV DNA positive in PBMC and serum of pregnant women are the risk factors for HBV intrauterine infection. Antepartum injection of HBIG over thrice can protect placentas from being infected by HBV to some extent. PBMC HBV DNA positive in pregnant women is probably the independent risk factor for PBMC HBV intrauterine infection in newborns.

[Effects of climate change on flax development and yield in Guyuan of Ningxia, Northwest China].

Based on variations of the annual mean temperature and precipitation analyzed using ob- servation data in Guyuan of Ningxia, the effects of climate change on the local flax developmental process and yield were investigated. The results showed that the annual mean temperature had an increasing trend (0.3 °C · (10 a)-1) and the annual precipitation had a decreasing trend (-20 mm · (10 a) -1) from 1957 to 2012. While the increasing trend of mean temperature during growing season of flax was more obviously than that of the annual temperature, the decreasing trend of precipitation during growing season was similar to that of annual precipitation. With temperature increasing and precipitation decreasing, the flax development rate was accelerated, resulting in the reduced growing period. Seedling stage was advanced 0.7 d with 1 °C increase in temperature during the period from sowing to seedling emergence. The duration from seedling emergence to two pairs of needles was shortened by 0.8 d with 1 °C increase in temperature and 0.1 d with 1 mm decrease in precipitation. Maturity stage was advanced 1.8 d with 1 °C increase in temperature and 0.1 d with 1 mm decrease in precipitation during the period from technical maturity to maturity. The flax development was accelerated because of temperature increasing and precipitation decreasing in the vegetative growth phase, which was one of the main causes of flax yield reduction year by year. Meanwhile, flower bud differentiation and pollination of flax were influenced by temperature increasing in the reproductive growth phase, which would affect the number of capsules and the seed setting rate per plant and lead to the decrease of flax yield. Therefore, adjusting plant structure and enlarging the planting area of late or middle-late variety were the important measures to reduce the effects of climate change on local flax production.

[Expression and immunological activity of an anti-CD3×VEGFR2 bispecific single-chain antibody].

AIM: To construct and express an anti-VEGFR2/anti-CD3 bispecific single-chain antibody (bscVEGFR2×CD3)and to identify its binding specificities to CD3 and VEGFR2. METHODS: The gene encoding anti-VEGFR2/anti-CD3 bispecific single-chain antibody was designed and synthesized. Bispecific single-chain antibody (bsc-Ab) DNA was subcloned into a eukaryotic expression vector pcDNA3.1(+), then transfected into Chinese hamster ovary (CHO) cells and stable expression cell lines were selected. Expressed Bsc-Ab was purified by His-tag affinity chromatography and confirmed by 120 g/L SDS-PAGE and Western blotting. Antigen binding activity of the bsc-Ab was analyzed by FACS. RESULTS: The plasmid DNA containing bispecific single-chain fragments were confirmed. BscVEGFR2×CD3 was secreted by CHO into the supernatant. Six stable expression cell lines were established. The molecular weight of bsc-Ab was correct indicated by SDS-PAGE and Western blotting. The bsc-Ab could specifically bind to CD3(+); jurkat cells and VEGFR2(+); A375 cells. CONCLUSION: An anti-VEGFR2/anti-CD3 bispecific single-chain antibody is successfully constructed and expressed, and the antibody has specific binding capacity to CD3 and VEGFR2.

Characterization of sodium dodecyl sulfate modified iron pillared montmorillonite and its application for the removal of aqueous Cu(II) and Co(II).

Anionic surfactant modified Fe-pillared montmorillonites were prepared by Fe-hydrate solution and sodium dodecyl sulfate (SDS) solution. These organo-inorgano complex montmorillonites were divided into three types (CM1, CM2 and CM3) depending on different intercalation processes. X-ray diffraction spectra, the Fourier transform infrared (FTIR) spectra were used to analyze the structure of the raw and modified montmorillonites. X-ray photoelectron spectra of the samples have been studied to determine spectral characteristics to allow the identification of Fe(III) hydroxide. The specific surface area of the host montmorillonite (M0) is 73.2m(2)/g, while for the modified montmorillonites it is 114.0m(2)/g, 117.2m(2)/g, and 115.8m(2)/g, respectively. The mesopore volumes of the montmorillonites decrease after modification. Ions of copper and cobalt were selected as adsorbates to evaluate the adsorption performance of each montmorillonite. The adsorption data was analyzed by both Freundlich and Langmuir isotherm models and the data was well fit by the Langmuir isotherm model. The adsorption was efficient and significantly influenced by metal speciation, metal concentration, contact time, and pH. Higher adsorption capacity of the modified montmorillonites were obtained at pH 5-6. The results of desorption indicated that the metal ions were covalently bound to the modified montmorillonites.