OBJECTIVETo reveal the transmembrane signal pathway participating in regulating neuron functions of treating Alzheimer's disease (AD) by acupuncture.

METHODSSAMP8 mice was used for AD animal model. The effect of acupuncture method for qi benefiting, blood regulating, health supporting, and root strengthening on the amount and varieties of transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice was detected using HPLC MS/MS proteomics method.

RESULTSCompared with the control group, acupuncture increased 39 transmembrane signal proteins from hippocampal lipid rafts in SAMP8 mice, of them, 14 belonged to ionophorous protein, 8 to G protein, 8 to transmembrane signal receptor, and 9 to kinase protein. Totally 3 main cell signal pathways were involved, including G-protein-coupled receptors signal, enzyme linked receptor signal, and ion-channel mediated signal. Compared with the sham-acupuncture group, acupuncture resulted in significant increase of kinase signal protein amount. From the aspect of functions, they were dominant in regulating synapse functions relevant to cytoskeleton and secreting neurotransmitters.

CONCLUSIONThe cell biological mechanism for treating AD by acupuncture might be achieved by improving synapse functions and promoting the secretion of neurotransmitters through transmembrane signal transduction, thus improving cognitive function of AD patients.

Acupuncture Therapy ; Alzheimer Disease ; metabolism ; Animals ; Disease Models, Animal ; Female ; Male ; Membrane Microdomains ; metabolism ; Mice ; Proteomics ; Signal Transduction ; Tandem Mass Spectrometry

Traditional processes are mostly adopted in traditional Chinese medicine (TCM) preparation production and the quality of products is mostly controlled by terminal. Potential problems of the production in the process are unpredictable and is relied on experience in most cases. Therefore, it is hard to find the key points affecting the preparation process and quality control. A pattern of research and development of traditional Chinese medicine preparation process based on the idea of Quality by Design (QbD) was proposed after introducing the latest research achievement. Basic theories of micromeritics and rheology were used to characterize the physical property of TCM raw material. TCM preparation process was designed in a more scientific and rational way by studying the correlation among enhancing physical property of raw material, preparation process and product quality of preparation. So factors affecting the quality of TCM production would be found out and problems that might occur in the pilot process could be predicted. It would be a foundation for the R&D and production of TCM preparation as well as support for the "process control" of TCMIs gradually realized in the future.
Drug Compounding ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; trends ; Quality Control ; Research ; standards ; trends ; Research Design ; standards ; Technology, Pharmaceutical ; instrumentation ; methods ; standards

Objective · To investigate whether the quality of embryos will result in biochemical pregnancy or arrest of embryo development in the freezing and thawing cycles of in-vitro fertiliazation-embryo transfer (IVF-ET). Methods · The clinical data of patients who accepted IVF-ET in Center of Reproductive Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from January 2015 to June 2016 were retrospectively studied. The data includes 115 cycles of biochemical pregnancy, 64 cycles of arrest of early embryonic development and 871 cycles of ongoing pregnancy after frozen thawed embryo transfer. We compared the embryo score on the third day after embryo transfer (D3), the blastocyst development rate and the blastocyst grade in the three groups. Results · There were no significant differences in the period of infertility, the age of the patients and their spouses, the endometrial thickness, the estrogen and progestogen levels of the day of transplantation among the three groups (P > 0.05). The scores of most frozen thawed embryos on D3 were from 6 to 8, and the scores were not statistically significant among the three groups (P > 0.05). The proportion of transplanted blastocyst on D5 was higher than that on D6 in the three groups, but there was no significant difference among the three groups (P > 0.05). There was no significant difference in the proportion of inner cell mass of blastocysts which were scored as Grade A&B or Grade C among the three groups. Nevertheless, in the arrest of early embryonic development group, the proportion (52.2%) of the trophoblast of blastocysts which were cored as Grade C was significantly higher than the proportion (35%) in biochemical pregnancy group and the proportion (29.3%) in ongoing pregnancy group (P<0.05). Conclusion · The quality of embryos is not necessarily related to biochemical pregnancy, but the score of trophoblastic may be related to the arrest of early embryo growth.

Objective To study the antigenicity of human thyrotropin receptor(hTSHR)amino terminus (amino acid 29～280)and its association with Graves' disease.Methods Total thyroid RNA was prepared from human normal thyroid tissue.RNA was then reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and the recombinant plasmid was named pcDNA3.1/hTSHR188～940bp. Balb/c mice were immunized with peDNA3.1/hTSHR188～940bp. The levels of serum thyroxin,anti-TSHR antibody(TRAb)and thyroid stimulating antibody(TSAb)were measured,and the pathological changes of thyroid tissue were also observed.Results A 753 bp fragment encoding hTSHR ectodomain amino end was obtained after PCR amplification.Confirmed by Hind Ⅲ restriction enzyme digestion and DNA sequencing,pcDNA3.1/hTSHR188～940bphad been constructed successfully,with the correct sequence and direction of hTSHR188～940bp.In the Balb/c mice treated with pcDNA3.1/hTSHR188～940bp,elevated TRAb in week 6(0.148±0.018)were observed compared with those at week o(0.106±0.006,P＜0.01),and kept a higher level till week 10(0.134±0.011,P＜0.01).T4 and TSAb index values were significantly increased in week 10.Serum T4 concentration increased from(41.02±7.97)μg/L in week 0 to(62.20±12.77)μg/L in week 10(P＜0.01);TSAb index values rose from 0.864±0.076 at week 0 to 1.392±0.615(P＜0.01).Thyroid pathological examination showed that proliferated thyroid follicular epithelial cells and foll icular eapacity increased.Inflammatory cells were occasionally found.Conclusions There are antigen epitopes in hTSHR ectodomain amino acid 29～280,which can stimulate the production of TSAb.And the latter induces hyperthyroidism and Graves' disease like manifestations.It suggests that hTSHR ectodomain amino acid 29～280 is closely associated with Graves' disease,and maybe one of important etiological factors leading to the disease.

Apyrase activities in some tissues and cells, such as peripheral vascular endothelial cells, have been reported, but these in endocardium endothelial cells have not been reported. The present study was to characterise the properties of bovine endocardium endothelial cells (BEEC)-associated apyrase. Apyrase activity was assayed by inorganic phosphate release, which could be inhibited concentration-dependently by NaN3, an apyrase inhibitor. NaF (20 mmol/L), another inhibitor of apyrase, also markedly inhibited the activity. EDTA or EGTA (1 mmol/L) could also inhibit the activity completely. However, the inhibitor for Na+/K+-ATPase, ouabain (3 mmol/L) did not affect the enzyme activity. BEEC apyrase activity was dependent on divalent cations (Ca2+ or Mg2+) and pH value.

Objective To construct the recombinant eukaryotic expression vector pRNAT-U6,1- siEdg4 which curries small interfering RNA(siRNA)of Edg4 and observe the silencing effect of Edg4 gene targeted siRNA in ovarian cancer cell line SKOV3.Methods The Edg4 gene-targeted hairpin siRNA sequence was designed according to the Edg4 sequence in Genbank,and the two complementary oligo nucleotide strands were synthesized and annealed and inserted into the pRNAT-U6.1 plasmid to build a recombinant Edg4 siRNA eukaryotic expression vector,which was sequenced and identified to contain the correct Edg4 siRNA sequence.The human ovarian carcinoma cell lines SKOV3 were transfeeted with the vector using lipofeetamine method.The efficiency of transfecting cells was observed with fluorescent microscope and the mRNA expression level of Edg4 gene was detected by real time quantitative PCR.The LPA levels in cell supernatants were detected using a biochemical method.And the apoptosis of SKOV3 cells induced by the vector was evaluated by flow cytometry.Results The recombinant eukaryotic expression vector was confirmed to contain correct Edg4 siRNA sequence by PCR and sequencing.After transfection large amounts of green fluorescence were seen in plasma and nuclei of SKOV3 cells and the positive cell rates were 64%.The expression level of Edg4 mRNA in transfeeted SKOV3 cell line was significantly decreased (0.05?0.01 vs 0.29?0.04,P

OBJECTIVETo study the effect of SiO(2) on the expression of platelet derived growth factor (PDGF) in human silicotic alveolar macrophages (AM) and human embryonic lung fibroblasts (HELF).

METHODSHuman alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to SiO(2) for 3, 6, 12, 18, 24 and 36 h. The cultured supernatant at 24 h was incubated with human embryonic lung fibroblasts for 6, 12, 18, 24, 36 and 48 h. The immunocytochemistry and Western blot were used to detect the level of expression of PDGF in lung fibroblasts and their supernatant respectively. (3)H-proline was used to detect the synthesis and secretion of collagen in HELF.

RESULTSThe expression of the PDGF in the supernatant of alveolar macrophages exposed to SiO(2) increased significantly and reached the peak at 24 h (average optical density: 0.282 +/- 0.019 vs 0.214 +/- 0.014, P < 0.01) with ELISA. The expression of PDGF in lung fibroblasts and their supernatant increased at different time (6, 12, 18, 24, 36 and 48 h) with immunocytochemistry and Western blot respectively when incubated with the cultured supernatant of silicotic AM exposed to SiO(2). The expression of PDGF was significantly different from the control group (P < 0.05). The synthesis and secretion of collagen in FB were increased markedly when incubated with the cultured supernatant of AM stimulated by SiO(2) compared with the control group.

CONCLUSIONSiO(2) may affect the expression of PDGF and synthesis of collagen through AM mediation and participate in the formation of lung fibrosis.

Cells, Cultured ; Collagen ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Macrophages, Alveolar ; drug effects ; metabolism ; Male ; Middle Aged ; Platelet-Derived Growth Factor ; metabolism ; Silicon Dioxide ; pharmacology

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; In Situ Nick-End Labeling ; Liver ; cytology ; drug effects ; ultrastructure

OBJECTIVETo investigate the effects of cis-combretastatin-A1 phosphate (cis-CA1P) on tumor cell proliferation, and its effects on the blood vessel formations.

METHODSMTT and IC50 values were used to assess the inhibitory effects of cis-CA1P on tumor cell proliferation. Chicken embryo chorioallantoic membrane and thoracic aorta annulations isolated from rats were used to investigate the effects of cis-CAIP on the blood vessel formation.

RESULTSCis-CA1P concentration-dependently inhibited the proliferations of several cancer cell lines, including human gastric carcinoma cell line MGC-803, human leukemic monocyte lymphoma cell line U937, human melanoma cell line A375, human colon cancer cell line HCT116, human breast carcinoma cell line MDA-MB-231, and human leukemia cell line K562. Cis-CAIP significantly decreased the formation of blood vessels in chicken embryo chorioallantoic membrane and in thoracic aorta annulations.

CONCLUSIONCis-CA1P inhibits cancer cell proliferation and prevents blood vessel formation.

Animals ; Aorta ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; drug effects ; Humans ; In Vitro Techniques ; Neovascularization, Pathologic ; prevention & control ; Phosphates ; pharmacology ; Rats ; Stilbenes ; chemistry ; pharmacology

OBJECTIVEThe present study was designed to investigate the influence of viral myocarditis on mental behavior of the children.

METHODSMentality and behavior of the 178 children suffering from viral myocarditis were examined with Achenbach Child Behavior Checklist (CBCL) edited by Gong Yao-xian in 1986 and 104 sex, age, education and achievement-matched children with pneumonia or upper respiratory infection were enrolled as controls.

RESULTSThe mentality and behavior of the children with viral myocarditis were distinctly abnormal. The abnormal rates of boys and girls suffering from acute and deferment viral myocarditis were evidently higher than those of control children (P < 0.01). The behavioral abnormalities of boys were hypochondria, social difficulties, unwell of body and attack. Whereas, the girls presented hypochondria, unwell of body, social flinch and disobeyed discipline, which was significantly different from the control children. The total and hypochondria cursory mark of the second check result of deferment boys were evidently higher than those of the first check (P < 0.05). The total cursory mark of the second check result of deferment girls was higher than that of the first check (P < 0.05) and evidently higher than that of the acute second check result (P < 0.01). The abnormal rates of mentality and behavior correlated positively with the age of children and they were associated with the severity of the illness.

CONCLUSIONViral myocarditis evidently affected the mentality and behavior of children, which should be paid great attention to.

Acute Disease ; Adolescent ; Adolescent Behavior ; psychology ; Case-Control Studies ; Child ; Child Behavior ; psychology ; Child Behavior Disorders ; etiology ; psychology ; Female ; Humans ; Hypochondriasis ; etiology ; psychology ; Juvenile Delinquency ; psychology ; Male ; Myocarditis ; psychology ; virology ; Neuropsychological Tests ; Severity of Illness Index ; Social Behavior ; Virus Diseases ; psychology