Astragalus polysaccharide has been widely used in food and medicinal industry owing to its health-promoting properties. In order to characterize better the relationship among molecular weight, structure-activity and activities, a simple method was used different concentration of ethanol including 30% (PW30), 50% (PW50), 70% (PW70), 75% (PW75), 80% (PW80) and 90% (PW90) to precipitate Astragalus polysaccharides into different molecular weight. As a result, PW90 showed smooth surface and the strongest antioxidant activity among these six fractions (P < 0.05). In conclusion, graded ethanol precipitation was a simple method to separate Astragalus polysaccharides into different molecular weight with different antioxidant activity fractions.
Antioxidants ; pharmacology ; Astragalus Plant ; chemistry ; Chemical Precipitation ; Ethanol ; chemistry ; Polysaccharides ; chemistry ; pharmacology

Tumor angiogenesis provides adequate oxygen and nutrition for tumor development and supports tumor growth and metastasis. Stromal cell derived factor 1 (SDF-1) and its receptor C-X-C motif chemokine receptor 4 (CXCR4) in pancreatic cancer microenvironment are involved in tumor growth such as promoting tumor cell proliferation, migration, and angiogenesis. In this study, anti-CXCR4 nanobody (CXCR4 Nb) and anti-programmed cell death ligand 1 (PD-L1) &CXCR4 bispecific nanobody (PX4 BsNb) were expressed in Escherichia coli system and purified by nickel column affinity chromatography. We investigated the anti-angiogenesis activity and mechanism of CXCR4 Nb by in vivo and in vitro experiments. Ethical approval was obtained for collection of human peripheral blood mononuclear cell (hPBMC) samples from the Local Ethics Committee of Shanghai Jiao Tong University. All animal experiments were approved by the Animal Ethic Committee of Shanghai Jiao Tong University. The results showed that CXCR4 Nb at 0.1 μmol·L^-1 could effectively inhibit the proliferation and migration of human umbilical vein endothelial cells (HUVEC) promoted by pancreatic stellate cells in vitro. CXCR4 Nb and PX4 BsNb at 0.3 mg·kg^-1 obviously decreased tumor angiogenesis and inhibited the tumor growth in NOD/SCID mice, the inhibitory rates were 28.8% and 36.1%, respectively. CXCR4 Nb significantly inhibited tumor growth and angiogenesis with great safety, which provides support for application of CXCR4 Nb and anti-angiogenesis therapy of pancreatic cancer.

OBJECTIVETo identify SNP in flos Lonicerae, and authenticate Lonicera japonica from its adulterants and the mixture by using bidirectional PCR amplification of specific alleles (Bi-PASA).

METHODSNP of L. japonica and its adulterants was identified by using ClustulW to align trnL-trnF sequences of the Lonicera genus from GenBank database. Bi-PASA primer was designed and the PCR reaction systems including annealing temperature optimized. Optimized result was performed in 84 samples to authenticate L. japonica, its adulterants and the mixture.

RESULTWhen the annealing temperature was 61 degrees C, DNA from L. japonica would be amplified 468 bp whereas PCR products from all of the 9 adulterants were 324 bp. The established method also can detect 5% of intentional adulteration DNA into L. japonica.

CONCLUSIONThe Bi-SPASA could authenticate L. japonica from its adulterants and the mixture.

Alleles ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; analysis ; genetics ; Flowers ; genetics ; Lonicera ; classification ; genetics ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; RNA, Transfer, Leu ; genetics ; RNA, Transfer, Phe ; genetics ; Reproducibility of Results ; Species Specificity

Increasing evidence has revealed that maternal cytomegalovirus (CMV) infection may be associated with neurodevelopmental disorders in offspring.Potential relevance between the placental inflammation and CMV-related autism has been reported by clinical observation.Meanwhile,abnormal expression of Toll-like receptor 2 (TLR2) and TLR4 in placenta of patients with chorioamnionitis was observed in multiple studies.IL-6 and IL-10 are two important maternal inflammatory mediators involved in neurodevelopmental disorders.To investigate whether murine CMV (MCMV) infection causes alterations in placental IL-6/10 and TLR2/4 levels,we analyzed the dynamic changes in gene expression of TLR2/4 and IL-6/10 in placentas following acute MCMV infection.Mouse model of acute MCMV infection during pregnancy was created,and pre-pregnant MCMV infected,lipopolysaccharide (LPS)-treated and uninfected mice were used as controls.At E13.5,E14.5 and E18.5,placentas and fetal brains were harvested and mRNA expression levels of placental TLR2/4 and IL-6/10 were analyzed.The results showed that after acute MCMV infection,the expression levels of placental TLR2/4 and IL-6 were elevated at E13.5,accompanied by obvious placental inflammation and reduction of placenta and fetal brain weights.However,LPS 50 μg/kg could decrease the IL-6 expression at E13.5 and E14.5.This suggests that acute MCMV infection during pregnancy could up-regulate the gene expression of TLR2/4 in placental trophoblasts and activate them to produce more proinflammatory cytokine IL-6.High dose of LPS stimulation (50 tg/kg) during pregnancy can lead to down-regulation of IL-6 levels in the late stage.Imbalance ofIL-6 expression in placenta might be associated with the neurodevelopmental disorders in progeny.

OBJECTIVETo probe into effective methods for treatment of ischemic cerebral infarction and specificity of acupoints in the acupuncture treatment.

METHODSThe rat model of middle cerebral artery occlusion (MCAO) was established with thread ligation according to Zea-Longa's method. They were divided into normal group, non-operation group, model control group, non-acupuncture group and acupuncture group, and the acupuncture group was divided into sham-acupuncture group and four Xingnao Kaiqiao groups (contain Shuigou group, Neiguan group, Chize group, Sanyinjiao group and Weizhong group). Then they were treated by acupuncture at "Shuigou" (GV 26), "Neiguan" (PC 6), "Chize" (LU 5), "Sanyinjiao" (SP 6), "Weizhong" (BL 40) and non-acupoints, 3 times/second, for 5 seconds. Cerebral blood blow (CBF) was used for assessment of the effect.

RESULTSAs compared with the model control group, in the non-acupuncture group CBF did not significantly change (P>0.05); compared with the non-acupuncture group, after acupuncture CBF was significantly increased in the Shuigou group and the Neiguan groups (all P<0.05), but did not significantly increased in the Chize group, Sanyinjiao group and Weizhong group (all P>0.05).

CONCLUSIONCBF has a tendency of spontaneous cure within 72 h after cerebral infarction in the MCAO rat; acupuncture can significantly improve CBF in the MCAO rat, so it is an effective method for treatment of ischemic stroke; among the acupoints in the "Xingnao Kaiqiao" needling method, "Shuigou" (GV 26) and "Neiguan" (PC 6) have obvious effect in improvement of CBF, indicating acupoint specificity of "Shuigou" (GV 26) and "Neiguan" (PC 6) in treatment of ischemic stroke.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Brain ; blood supply ; Brain Ischemia ; therapy ; Cerebral Infarction ; therapy ; Disease Models, Animal ; Humans ; Male ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of acupoint Shuigou (GV 26) and non-acupoint on inhibiting the neuronal necrosis induced by the middle cerebral artery obstruction (MCAO) in rats through the morphological observation.

METHODSForty two healthy adult male Wistar rats were randomly divided into a normal group, false-ope ration group, model control group, non-acupuncture group, Shuigou-acupuncture group and non-acupoint acupuncture group, 7 rats in each group. Besides the normal and false-operation groups, the experimental modal of MCAO was established in the other groups by using the Zea-Longa thread method. Acupoint Shuigou (GV 26) and non-acupoint (below the costal region) were selected respectively in the Shuigou-acupuncture group and non-acupoint acupuncture group for puncturing at 180 times/min for 5 seconds. Its effect was estimated by measuring the rate of the neuronal necrosis at the level of the light microscope and the degree of the neuronal necrosis at the level of the electron microscope.

RESULTS(1) At the level of the light microscope, comparing with the model control group (0.66 +/- 0.18), of the neuronal necrosis there were no significant differences in the rate of neuronal necrosis in the non-acupuncture group (0.67 +/- 0.34) and non-acupoint acupuncture group (0.59 +/- 0.11) (both P > 0.05), while it was significantly decreased in the Shuigou-acupuncture group (0.200 +/- 12) (P < 0.05). It indicates that no intervention and wrong (non-acupoint) intervention could light the damage of the neurons, however, only right intervention (Shuigou, GV 26) can obviously decrease the rate of the neuronal necrosis, showing with the specificity of acupoint. (2) At the level of the electron microscope, the ultrastructure of the involved neurons in the non-acupuncture group and non-acupoint acupuncture group is similar to that of the model control group, showing with the serious edema and structure damage. In contrast, the ultrastructure of the involved neurons in the Shuigou-acupuncture group is similar to that of the normal group and false-operation group, showing with light neuronal damage.

CONCLUSIONAcupuncturing Shuigou (GV 26) could protect the involved neurons and inhibit the neuronal necrosis induced by the MCAO in rats, but acupuncturing the non-acupoint couldn't, which imply that there is the specificity of acupoint in the acupuncture.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Brain Ischemia ; pathology ; physiopathology ; therapy ; Disease Models, Animal ; Humans ; Male ; Necrosis ; Neurons ; pathology ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar

PURPOSE: The purpose of this study was to develop and preliminarily evaluate the reliability and validity of the Symptom-Management Self-Efficacy ScaleeBreast Cancer (SMSES-BC) related to chemotherapy. METHODS: The study included three stages. This paper presents the results of stage 2 and stage 3. In total, 34 items in the SMSES-BC were found during stage 1 from qualitative findings, a literature review, and expert suggestions; the 34 items were used for the psychometric properties test. Test-retest reliability and Cronbach alpha were assessed in the first sample, which included 45 participants for the pilot test (stage 2). The second sample, which included 152 patients, was used to assess the construct validity and concurrent validity (stage 3). RESULTS: The pilot test results revealed a test-retest reliability of .73 (p < .001) and Cronbach alpha coefficient of .96 for the total scale. Three factors (managing chemotherapy-related symptoms, acquiring problem solving, and managing emotional and interpersonal disturbances) were identified from exploratory factor analysis. Correlation coefficient r was .40 (p < .001), which supported the association between SMSES-BC and the General Self-Efficacy Scale for concurrent validity. CONCLUSIONS: The study results demonstrate acceptable reliability and validity for the SMSES-BC that was developed for measuring symptom-management self-efficacy related to chemotherapy for patients with breast cancer. This study suggests further research to validate the construct of the SMSES-BC.
Adult ; Aged ; Antineoplastic Agents/*therapeutic use ; Breast Neoplasms/*drug therapy/*psychology ; Drug Therapy/*psychology ; Factor Analysis, Statistical ; Female ; Humans ; Middle Aged ; Patients/*psychology ; Pilot Projects ; Psychometrics ; Reproducibility of Results ; Self Care/*psychology ; Self Efficacy ; Surveys and Questionnaires ; Taiwan

OBJECTIVETo screen the proteins interacting with FXR1P for functional investigation of FXR1P.

METHODSThe yeast strain AH109 transformed with the recombinant expression vector pGBKT7/FXR1 was mated with the yeast strain Y187 pretransformed with human fetal brain cDNA library. The positive clones were screened and identified by sequence analysis.

RESULTSThe recombinant expression vector pGBKT7/FXR1 was constructed successfully. Five proteins binding to FXR1P were screened from human fetal brain cDNA library using the yeast two-hybrid system, including CMAS, FTH1, GOLGA4, HSD17B1 and CSH1.

CONCLUSIONSThese results provide new clues for investigating the biological functions of FXR1P and the pathogenesis of Fragile X syndrome.

Autoantigens ; genetics ; metabolism ; Estradiol Dehydrogenases ; genetics ; metabolism ; Ferritins ; genetics ; metabolism ; Gene Library ; Humans ; Membrane Proteins ; genetics ; metabolism ; Protein Binding ; Protein Interaction Domains and Motifs ; genetics ; RNA-Binding Proteins ; genetics ; metabolism ; Two-Hybrid System Techniques

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection