Objective To explore the residual undifferentiated mouse embryonic stem cells (ESCs) in embryoid bodies. Methods Mouse R1 and Oct-4-GFP transgenic ESCs were firstly cultured in suspension to form embryoid bodies (EBs). Twenty days later, EBs were digested into single cells and then re-plated in standard ESC culture condition. The morphology of residual undifferentiated cells in EBs was observed, and surface makers and in vitro redifferentiation potency of residual cells were examined by flow cytometry and immunofluoreseent staining. The residual cells were expanded and subcutaneously injected into nude mice, and the specimens were harvested from the injection site for histological analysis 6 weeks after injection. Results There were residual undifferentiated ESCs in EBs differentiated for 20 days, which displayed clonal morphology and expressed undifferentiated cell markers of ESCs, including SSEA1, CD31, CD9 and Oct-4. The cells could be differentiated to form EBs again, and could be re-expanded from secondary EBs. The residual cells were able to form teratoma at the injection site, and mature endoderm, mesoderm and ectoderm tissues could be found in teratoma tissues. Conclusion There are residual undifferentiated ESCs after differentiation of ESCs into EBs. The residual ESCs can differentiate again in vitro and in vivo, and can residue again in the in vitro differentiation.

italic>Ardisia crispa (Thunb.) A. DC. is a traditional Miao medicinal herb with significant therapeutic effects in the treatment of sore throat, tonsillitis, edema of nephritis and bruising and rheumatism, etc. Ardisia crispa var. amplifolia and Ardisia crispa var. dielsii are varieties of A. crispa. A. crispa var. amplifolia and A. crispa var. dielsii are controversial in terms of species evolutionary relationships and taxonomic identification. In this study, we sequenced the whole genome sequences of A. crispa var. amplifolia and A. crispa var. dielsii chloroplasts using Illumina platform, assembled, annotated and characterized them, compared the structural features and degree of variation among chloroplast genomes using bioinformatics methods, and also downloaded constructing phylogenetic trees to analyze the phylogenetic relationships of chloroplasts in Primulaceae and Myrsinaceae using whole genome sequence information. The results showed that the complete chloroplast genome sequences of A. crispa var. amplifolia and A. crispa var. dielsii were 156 749 bp and 156 748 bp in length, with 132 genes annotated, including 87 protein-coding genes; the codon preference of A/U was greater than that of G/C; The differences in the coding regions of rps15 and rpoB genes in the comparative genome analysis can be used as loci for molecular identification of the two species; the differences in the coding regions of ycf1, ycf2, rpoC1, ycf3, petD and rpl16 genes in the chloroplast genome compared with those of the same genus can be used as loci for identification of the genus. In the phylogenetic results, A. crispa var. amplifolia and A. crispa var. dielsii were clustered together with 100% support, indicating that they are closely related. In this research, we analyzed the chloroplast genome structure and phylogenetic relationships of A. crispa var. amplifolia and A. crispa var. dielsii, providing an important theoretical basis for their molecular identification, genetic variation, breeding and phylogenetic analysis.

Child, Preschool ; China ; Female ; Hepatitis A ; immunology ; prevention & control ; Hepatitis A Antibodies ; blood ; Hepatitis A Vaccines ; adverse effects ; immunology ; Humans ; Infant ; Male ; Safety ; Sampling Studies ; Vaccination ; Vaccines, Inactivated ; adverse effects ; immunology

OBJECTIVETo study eleven organophosphorus insecticides residuals in four kinds of Chinese crude drugs.

METHODThe organophosphorus insecticides were extracted with dichloromethane and cleaned-up with a mixture of Celite 545-activated carbon. The extracts were analyzed by gas chromatography equipped with a flame photometric detector (FPD).

RESULTAnalysis of fortified Chinese crude drug showed that the average recoveries ranged from 77.5% -112.3% at three different levels, the RSDs were below 10% (n = 4). Trace organophosphorous pesticide residues were found in samples of Rhizoma Atractylodis Macrocephalae and Flos Chrysanthemi.

CONCLUSIONA method was established for determination multi-residues in Rhizma Atractylodis Macrocephalae, Radix Curcumae, Bulbus Fritillariae Thunbergii and Flos Chrysanthemi. It provides a method for the risk assessment of organophosphorous pesticide in Chinese crude drugs.

Atractylodes ; chemistry ; Chromatography, Gas ; Chrysanthemum ; chemistry ; Curcuma ; chemistry ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Fritillaria ; chemistry ; Organophosphorus Compounds ; analysis ; Pesticide Residues ; analysis ; Plants, Medicinal ; chemistry

OBJECTIVETo explore the effect of apigenin on semen parameters in male mice.

METHODSTotally 100 healthy male mice of Kunming strain were randomly divided into 5 groups according to the body weight: negative control, solvent control, low-dose apigenin, median-dose apigenin and high-dose apigenin, the latter three groups given intragastric apigenin at a fixed time every day for 7 and 14 days. At 35 days after the first medication, all the mice were killed and detected for the sperm motion parameters by computer aided sperm analysis (CASA).

RESULTSThere were no statistically significant differences in sperm motion parameters, density and motility between the negative control and the three apigenin groups after 7-day medication. At 14 days, the high-dose apigenin group showed remarkable decreases in average path velocity (VAP), straight line velocity (VSL), straightness (STR), wobbliness (WOB), the percentage of grade b sperm and sperm motility, and a significant increase in beat cross frequency (BCF) as compared with the negative control group (P < 0.05).

CONCLUSIONApigenin affects sperm motility in male mice to a certain extent.

Animals ; Apigenin ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Semen ; drug effects ; Sperm Motility ; drug effects

Objective To explore the effect of Internet addiction on adolescent's attention.Methods Neuropsychological evaluations of attention function were done separately in 18 adolescents who met the diagnostic criterion for Internet addiction and in other 18 ones without Intemet addition tendency as a control group. With auditory and visual oddball paradigms, and stimulated by standard,target and novel stimuli, EEG was recorded and analyzed to get the event-related potential P300 and compare the latency and amplitude of P3a and P3b between the 2 groups. Results Compared with the controls, the attention of Internet addiction group was decreased significantly in neuropsychological evaluations. The latency of P3a potentials induced by novel stimulus was much shorter and the amplitude of it was higher in addiction group than in control group, but the latency of P3b potentials generated by target stimulus was prolonged and the amplitude decreased obviously. Conclusions Intemet addiction can cause damage to adolescent's attention function. There is a correlation between the attention impairment and the change of P300 potentials.

OBJECTIVETo investigate the levels of cytokines related to T-helper (Th) 17 cells in serum and signal transducers in the psoriatic lesions of patients with psoriasis vulgaris of blood-heat syndrome (BHS) and blood-stasis syndrome (BSS).

METHODSSixty patients with psoriasis vulgaris were divided into the BHS and BSS groups according to the syndrome differentiation of Chinese medicine (CM). Ten healthy subjects were considered as the control group. Cytokine levels of interleukin (IL)-17, IL-23 and IL-6 in serum were determined by enzyme-linked immunosorbent assay. Expression levels of signal transducer and activator of transcription 3 (STAT3), p38-mitogen-activated protein kinase (MAPK) and STAT6 in the psoriatic lesions were determined using immunohistochemistry (IHC), Western blot, and real-time quantitative reverse transcription polymerase chain reaction, respectively.

RESULTSProduction of IL-17, IL-23 and IL-6 in the BHS group and BSS group were significantly increased compared with those in the control group (P<0.05). Levels of IL-17 and IL-23 in the BHS group were higher than those in the BSS group (P<0.05). Compared with the control group, IHC positive expressions and protein expressions of STAT3 and p38-MAPK, and the STAT3 mRNA expressions in the BHS and BSS groups were significantly higher (P<0.05 or P<0.01). The protein expression of STAT3 in the BHS group was significantly higher than that in the BSS group (P<0.05).

CONCLUSIONSCytokines in serum and signal transducers in the psoriatic lesions alter with various CM syndromes of psoriasis. The results provide scientific basis for the treatment based on syndrome differentiation of CM in treating psoriasis vulgaris.

Adult ; Female ; Gene Expression Regulation ; Humans ; Immunohistochemistry ; Interleukin-17 ; blood ; Interleukin-23 ; blood ; Interleukin-6 ; blood ; Male ; Psoriasis ; blood ; enzymology ; genetics ; immunology ; RNA, Messenger ; genetics ; metabolism ; STAT3 Transcription Factor ; genetics ; metabolism ; STAT6 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; Syndrome ; Th17 Cells ; immunology ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism