Animals ; Cell Differentiation ; Cytokines ; metabolism ; Disease Progression ; Epithelial-Mesenchymal Transition ; Extracellular Matrix ; metabolism ; Fibroblasts ; metabolism ; pathology ; Hepatic Stellate Cells ; metabolism ; pathology ; Humans ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; etiology ; metabolism ; pathology ; Stem Cells ; metabolism ; pathology

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

To develop a cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy, we prepared the AVPI-LMWP/pTRAIL self-assembled complexes containing a therapeutic combination of peptide drug AVPI and DNA drug TRAIL. The chimeric apoptotic peptide AVPI-LMWP was synthesized using the standard solid-phase synthesis. The cationic AVPI-LMWP could condense pTRAIL by electrostatic interaction. The physical-chemical properties of the AVPI-LMWP/pTRAIL complexes were characterized. The cellular uptake efficiency and the inhibitory activity of the AVPI-LMWP/pTRAIL complexes on tumor cell were also performed. The results showed that the AVPI-LMWP/pTRAIL complexes were successfully prepared by co-incubation. With the increase of mass ratio (AVPI-LMWP/DNA), the particle size was decreased and the zeta potential had few change. Agarose gel electrophoresis showed that AVPI-LMWP could fully bind and condense pTRAIL at a mass ratio above 15:1. Cellular uptake efficiency was improved along with the increased ratio of W(AVPI-LMWP)/WpTRAIL. The in vitro cytotoxicity experiments demonstrated that the AVPI-LMWP/pTRAIL (W:W = 20:1) complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRAIL complexes on inhibition of HeLa cell growth. Our studies indicated that the AVPI-LMWP/pTRAIL co-delivery system could deliver plasmid into HeLa cell and induce tumor cell apoptosis efficiently, which showed its potential in cancer therapy using combination of apoptoic peptide and gene drugs.
Antineoplastic Agents ; chemistry ; Cell-Penetrating Peptides ; chemistry ; DNA ; chemistry ; Drug Delivery Systems ; HeLa Cells ; Humans ; Neoplasms ; drug therapy ; Particle Size ; Plasmids

To study the chemical constituents of Cymbopogon citratus, isolation and purification of constituents were carried out on silica gel, Sephadex LH-20 and prepatative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Eight compounds were isolated and identified as 3beta-methoxy lanosta-9(11)-en-27-ol (1), 3beta-hydroxylanosta-9 (11)-en (2), (24S) -3beta-methoxylanosta-9(11), 25-dien-24-ol (3), 8-hydroxyl-neo-menthol (4), (2E)-3,7-dimethyl-2,7-octadiene-1, 6-diol (5), (+)-citronellol (6), 7-hydroxymenthol (7) and ethyl nonadecanoate(8). Compounds 1 is a new one. Compounds 2-3 are obtained from C. citratus for the first time.
Cymbopogon ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene, to reveal its mutation features, and to establish a basis for genetic counseling and prenatal gene diagnosis of Hunter syndrome.

METHODSUrine glycosaminoglycans (GAGs) assay, PCR and DNA sequencing were performed to detect mutation of IDS gene of the patient and his parents.

RESULTSThe result showed that the patient was: DS(++), HS(++), KS(-), CS(-), and that both of his parents were negative. A frame-shift deletion mutation (1062 del 16) was identified in exon 7 of the patient's IDS gene. His parents' genotypes were normal.

CONCLUSIONThe patient's mutation was not inherited by his parents but a novel one. The mutation probably altered the primary structure and tertiary structure of IDS enzyme protein remarkably and lowered the activity of IDS enzyme greatly. Therefore it is supposed to be the direct cause of the disorder.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child, Preschool ; Female ; Glycoproteins ; genetics ; urine ; Humans ; Male ; Mucopolysaccharidosis II ; enzymology ; genetics ; urine ; Mutation ; genetics

Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg^-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between C_max and AUC_0-5d in the two groups. The liquid preparation was the reference preparation, with C_max ratio of 101.6% and AUC_0-5d ratio of 101.9%, the 90% confidence interval of C_max was 79.42%-129.92%, and the 90% confidence interval of AUC_0-5d was 85.72%-121.18%. These results suggested that different dosage forms of MIL50 had certain differences in the changes of blood drug concentration in rhesus monkeys.

Objective To establish and evaluate the predictive value of the risk prediction model for lung infection within postoperative 1 year in kidney transplant recipients. Methods Clinical data of 197 kidney transplant recipients were retrospectively analyzed. All recipients were divided into the infection group (n=42) and non-infection group (n=155) according to the incidence of lung infection within postoperative 1 year. The incidence and risk factors of lung infection after kidney transplantation were analyzed. Risk prediction model was established by multiple logistic regression analysis. Forty-five kidney transplant recipients who met the inclusion criteria, including 8 cases in the infection group and 37 cases in the non-infection group, were selected to verify the predictive effect of the established model. Results The incidence of lung infection within 1 year after kidney transplantation was 21.3% (n=42), including 38 cases (90%) of pneumonia severity index (PSI) class Ⅰ, 1 case (2%) of PSI class Ⅲ and 3 cases (8%) of PSI class Ⅴ. Lung infection occurred within 1 month after operation in 13 cases, within postoperative 2-6 months in 22 cases and after postoperative 6 months in 7 cases. Nineteen recipients were diagnosed with bacterial infection, 7 cases of fungal infection, 10 cases of viral infection and 6 cases of mixed infection. Smoking history, diabetes mellitus history, pulmonary disease history and albumin level of < 35 g/L were the independent risk factors for lung infection after kidney transplantation (all P < 0.05). The equation of risk prediction model for postoperative lung infection in kidney transplant recipients was logit (lung infection within postoperative 1 year in kidney transplant recipients)=-1.891+1.063×smoking history (yes=1, no=0)+1.398×diabetes mellitus history (yes=1, no=0)+1.732×pulmonary disease history (yes=1, no=0)+1.269×albumin level (< 35 g/L=1, ≥35 g/L=0). The area under the curve (AUC) of receiver operating characteristic (ROC) was 0.788, the sensitivity was 0.786, the specificity was 0.645, and the Youden index was 0.431, respectively. Hosmer-Lemeshow goodness-of-fit test demonstrated that the predicted value of this model yielded relatively high consistency with the observed value. The AUC in the verification group was 0.834. Hosmer-Lemeshow goodness-of-fit test validated high degree of calibration of this model. Conclusions The risk prediction model, consisting of smoking history, diabetes mellitus history, pulmonary disease history and albumin level as predictors, may effectively predict the incidence of lung infection within postoperative 1 year in kidney transplant recipients.

Objective:Response surface methodology was used to optimize the optimal extraction conditions of red ginseng polysaccharide and to study the antioxidant activity of red ginseng polysaccharide.Methods:Supercritical CO 2 extraction method was used to extract polysaccharides from red ginseng. The effects of solid-liquid ratio, extraction time, extraction temperature and extraction pressure on the extraction of polysaccharides from red ginseng were investigated. Box-behnken Design method was used to optimize the extraction process of red ginseng polysaccharide, and Logit method was used to calculate the semi-inhibitory concentration of red ginseng polysaccharide on DPPH clearance (half maximal inhibitory concentration, IC 50). Results:The optimal extraction conditions were as follows: extraction temperature 61.12 ℃, extraction pressure 20.64 MPa, extraction time 128.37 min, solid-liquid ratio 1∶25.61 g/ml, and the extraction yield of red ginseng polysaccharide was 36.89%. The results of three groups of repeatability tests showed that the relative error of polysaccharide yield of red ginseng was in the range of 5%. When the mass concentration of red ginseng polysaccharide was 25 μg/ml, it had better antioxidant activity and IC 50 was 10.97 μg/ml. Conclusion:The optimized extraction conditions of red ginseng polysaccharide were reasonable and reliable, and the antioxidant activity of red ginseng polysaccharide was strong, which could provide reference for the follow-up research.

Objective: To explore the effect of herb-partitioned moxibustion (HPM) on tight junctions (TJs) of intestinal epithelial cells in Crohn disease (CD) mediated by tumor necrosis factor-α (TNF-α)-nuclear factor kappa B (NF-κB)-myosin-light- chain kinase (MLCK) pathway. Methods: Forty-eight male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an HPM group and a mesalazine (MESA) group, with 12 rats in each group. Trinitrobenzene sulfonic acid (TNBS) was administered to establish CD models. When the model was confirmed a success, the HPM group rats were treated with HPM at Tianshu (ST 25) and Qihai (CV 6), while the MESA group rats were given MESA solution by lavage. When the intervention finished, the colonic epithelial tissues were separated, purified and cultured in each group to establish the intestinal epithelial barrier model in vitro, and TNF-α was added (100 ng/mL) in the culture medium and maintained for 24 h to establish an increased epithelial permeability model. Transepithelial electrical resistance (TEER) was used to examine the permeability of the barrier; Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway; immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium. Results: After TNF-α induction, compared with the MC+TNF-α group, the TEER value increased significantly in the HPM+TNF-α and MESA+TNF-α groups (both P<0.001); the expressions of nuclear factor kappa B (NF-κB) p65, MLCK, myosin light chain (MLC), tumor necrosis factor receptor-associated factor 6 (TRAF6) and receptor interaction protein-1 (RIP1) decreased significantly (P<0.01 or P<0.05), and the expression of zinc finger protein A20 (A20) increased significantly (P<0.01); the expressions of occludin, claudin-1, zonula occludens protein 1 (ZO-1) and F-actin also increased significantly (all P<0.01). Compared with the MESA+TNF-α group, the expressions of MLC, occludin, claudin-1, ZO-1 and F-actin increased significantly in the HPM+TNF-α group (P<0.01 or P<0.05). Conclusion: HPM can protect or repair the damage of intestinal epithelial barrier in CD rats, which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.

Autoimmune Diseases ; diagnosis ; genetics ; therapy ; Humans ; Liver Diseases ; diagnosis ; genetics ; therapy