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

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

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.

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

OBJECTIVESIn order to understand the effect of long term consumption of mineral-fortified dephytinized soy milk powder on skeletal health in children, biological effects of mineral-fortified dephytinized soy milk powder, dephytinized mineral-unfortified soy milk powder and fresh mineral-fortified cow milk were compared.

METHODSTotally, 90 boys aged 12 to 14 years were involved in the study and were given daily with mineral-fortified dephytinized soy milk powder, dephytinized soy milk powder and fresh fortified cow milk, respectively, for two months. The blood samples were collected from all the subjects for measuring activity of serum bone specific alkaline phosphatase and serum level of oesteocalcin, and the urine samples were collected for determining urine C-terminal telopeptides of type I collagen (CTx).

RESULTSAfter two-month intervention, serum level of oesteocalcin increased significantly in the all groups, (16.42 +/- 11.25) micro g/L for the group with fortified soy milk powder, (18.86 +/- 11.57) microg/L for the group with unfortified soy milk powder and (13.81 +/- 10.06) microg/L for the group with fortified cow milk, respectively. The activity of bone specific alkaline phosphatase significantly decreased in the all groups, by (0.28 +/- 0.40) micromol x s(-1) x L(-1) for the group with fortified soy milk powder, (0.26 +/- 0.35) micromol x s(-1) x L(-1) for the group with unfortified soy milk powder, and (0.17 +/- 0.28) micromol x s(-1) x L(-1) for the group with fortified cow milk, respectively. As well as, urine level of CTx decreased significantly, by (349.1 +/- 380.3) mg/L for the group with fortified soy milk powder, (260.4 +/- 412.4) mg/L for the group with unfortified soy milk powder, and (354.4 +/- 404.7) mg/L for the group with fortified cow milk, respectively.

CONCLUSIONSLong term consumption of mineral-fortified and dephytinized soy milk powder could improve bone mass retention and calcium nutrition status efficiently in children.

Adolescent ; Bone Density ; drug effects ; physiology ; Child ; Dietary Supplements ; Food, Fortified ; Humans ; Male ; Minerals ; pharmacology ; Phytic Acid ; metabolism ; Soybeans ; chemistry

OBJECTIVESIn order to understand the absorption and utilization of calcium, iron and zinc in cow milk, soy milk powder and dephytinized soy milk powder in the children and adolescents, and to lay a foundation for the improvement of protein and mineral nutrition in Chinese residents with popularizing consumption of soy milk powder.

METHODSTotally, 57 boys aged 12 to 14 years were selected and divided into three groups, matched by age, hemoglobin concentration, height and weight. The boys were given 220 ml of fortified and dephytinized soy milk, soy milk or cow milk, respectively, containing calcium 270 mg, iron 4 mg and zinc 4 mg, which were labeled with stable isotopes (44)Ca, (58)Fe and (70)Zn in a single serving size, and their feces were labeled with brilliant blue and dysprosium. Feces specimens were collected for all the subjects and absorption rate of calcium, iron and zinc were measured for all the children who took fortified and dephytinized soy milk, soy milk or cow milk, respectively. Calcium in feces was measured with heat ionized mass spectrometry, and recovery of (58)Fe and (70)Zn in feces and content of dysprosium were measured with induction coupling iso-ionic mass spectrometry, and then absorption rates of iron and zinc were calculated and adjusted by the recovery rate of dysprosium.

RESULTSIron absorption rate was (6.7 +/- 3.8)% in soy milk powder group, (15.5 +/- 9.2)% in the cow milk group and (20.6 +/- 7.3)% in dephytinized soy milk powder group, respectively. Calcium absorption rate was (43.5 +/- 10.7)%, (64.2 +/- 11.4)%, and (50.9 +/- 6.6)% in the three groups, respectively. Absorption rate of zinc was (11.3 +/- 6.5)%, (31.2 +/- 10.4)% and (20.1 +/- 7.4)%, respectively. Significant increase in absorption of calcium, iron and zinc was observed in the groups with fortified and dephytinized soy milk powder, as compared with those consuming nondephytinized soy milk powder. Absorption rate of calcium and zinc was significantly higher in the group with cow milk than that in the group with dephytinized and fortified soy milk powder, and iron bioavailability was lower in the group with cow milk than that with dephytinized soy milk powder, with no significant difference. Absorption rates of calcium, iron and zinc were higher in children of China due to their long term adaptation to dietary intake of lower mineral and protein, and higher fiber.

CONCLUSIONSBioavailability of calcium, iron and zinc in soy milk powder could be increased by dephytinized treatment for it.

Absorption ; drug effects ; physiology ; Adolescent ; Calcium ; metabolism ; Child ; Food, Fortified ; Humans ; Iron ; metabolism ; Male ; Minerals ; metabolism ; Phytic Acid ; pharmacology ; Soybeans ; chemistry ; Zinc ; metabolism

OBJECTIVETo investigate the factors affecting clinical therapeutic effect on acute tetramine poisoning.

METHODSUsing Logistic regression to analyze the relationships among the degree of tetramine poisoning, time of onset, time of admission, exposure history, sex, age, unithol, gastric lavage, etc with the death of poisonded patients.

RESULTSThe fatality rate of patient with tetramine poisoning who got gastric lavage was less than that who did not (5.85% vs 38.00%, P < 0.01). In patients who got gastric lavage, the fatality rates were increased with the degree of tetramine poisoning (control: 0%, mild poisoning: 3.07%, severe poisoning: 9.14%, P < 0.05). There was no significant difference in fatality between using unithol and not using patients (7.22% vs 8.25%, P > 0.05).

CONCLUSIONUnithol has no significant influence of clinical therapeutic effect on tetramine poisoning patients and dose not reduce the fatality rate of patient with tetramine poisoning, but gastric lavage and the degree of tetramine poisoning do. Logistic regression analysis showed that gastric lavage is the main factor affecting the therapeutic effect on tetramine poisoning.

Acute Disease ; Adolescent ; Adult ; Antidotes ; therapeutic use ; Bridged-Ring Compounds ; poisoning ; Child ; Female ; Gastric Lavage ; methods ; Humans ; Insecticides ; poisoning ; Logistic Models ; Male ; Poisoning ; mortality ; therapy ; Retrospective Studies ; Survival Rate ; Time Factors ; Treatment Outcome ; Unithiol ; therapeutic use

OBJECTIVETo investigate the relationship between the helicobacter pylori (HP) infection and the genetic instability of mitochondrial DNA (mtDNA) in human gastric adenocarcinoma epithelial cells (AGS).

METHODSAfter treated with extracts of HP11638 (CagA+, VacA+) or Hp11638 mutant strain (CagA+, VacA-), AGS cells were collected, and mitochondrial DNA was extracted and Cox-I, Cox-II, Cox-III, ATPase6, ATPase8 and Cytb genes and the D-Loop region were amplified by PCR and then sequenced.

RESULTSThe mutation rates of the mtDNA in AGS cells were correlated with the extracts of the two HP strains in a concentration- and time-dependent manner. But the mtDNA mutation rate in AGS cells treated with the HP11638 extract was higher than that treated with the Hp11638 mutant extract. Total of 616 mutations in D-Loop region were detected, including 489 point mutations, 81 insertions and 46 deletions. Among them, 70.9% (437/616) belonged to GC to AT and AT to GC transition. Seventeen out of 20 (85%) AGS cells treated with extract of HP had mutations in 303PolyC, 16184PolyC and 514CA regions of mtDNA D-Loop. No mutation was detected in Cox-I, Cox-II, Cox-III, ATPase6 and ATPase8 genes, three point mutations were found in the Cytb gene.

CONCLUSIONHP can cause the accumulation of mutations in mtDNA, in particular, in the D-Loop region, and the VacA participated in the process.

Antigens, Bacterial ; pharmacology ; Base Sequence ; DNA, Mitochondrial ; drug effects ; genetics ; Endothelial Cells ; drug effects ; pathology ; Helicobacter Infections ; complications ; Helicobacter pylori ; chemistry ; Humans ; Mutation ; Stomach ; pathology

OBJECTIVETo investigate the effect and regulation of extracellular signal-regulated kinase (ERK1/2) signaling pathway on the expression of transforming growth factor-beta1 (TGF-beta1) in human embryonic lung fibroblasts induced by SiO2.

METHODSHuman alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and in the presence or absence of SiO2 (50 ug/ml) exposition for 18h, and then the conditioned supernatants were used to incubate HELF. The expressions of TGF-beta1, of the HELF acted with the conditioned AM supernatant fluid were detected with the immunocytochemistry method after treatment with PD98059 of inhibitor of ERK.

RESULTSThe expression of TGF-beta1 in HELF of the SiO2 treatment group (OD value is 0.322 7 +/- 0.023 8) exceed blank group (OD value is 0.163 7 +/- 0.019 6) and AM control group (OD value is 0.240 6 +/- 0.022 5) by the immunocytochemistry method. But the expression of TGF-beta1 had reduction in some extent in the PD98059 intervention group (OD value is 0.271 1 +/- 0.022 9). The values were statistically different (P < 0.05).

CONCLUSIONERK inhibitor PD98059 have inhibition effect on the expression of transforming growth factor-beta1 and expression of cytokine of human embryonic lung fibroblasts stimulated by SiO2. The study indicate that the proliferation and collagen production of HELF activated by SiO2 are mediated by ERK/MAPK signal pathway in some extent. PD98059 may antagonizes silica-induced lung fibrosis by inhibiting the expression of transforming growth factor-beta1.

Bronchoalveolar Lavage Fluid ; cytology ; Cells, Cultured ; Fibroblasts ; metabolism ; Flavonoids ; pharmacology ; Humans ; Lung ; cytology ; metabolism ; MAP Kinase Signaling System ; Macrophages, Alveolar ; cytology ; metabolism ; Male ; Middle Aged ; Silicon Dioxide ; adverse effects ; Silicosis ; metabolism ; Transforming Growth Factor beta1 ; metabolism