In order to identify the chemical constituents of Yushu tablets comprehensively, we studied the chemical constituents of CHCl3 extract from Yushu tablets by the ultra performance liquid chromatography-electrospray ionization-ion trap-time of flight mass spectrometry (UPLC-ESI-IT-TOF/MS). It showed that there were more than 100 compounds separated, and forty-nine peaks among these were identified on the basis of high resolution mass spectrometry data and literature data reported. Determination of twelve peaks was further confirmed by standard substances. These components assigned to the different plant sources mainly included phenylpropanoids, triterpenoids, quinones and m-trihydroxybenzene compounds. By analyzing the chemical components of CHCl3 extract from compound Chinese medicine Yushu tablets comprehensively, this study provided the foundation for studying chemical components, pharmacodynamic substance and quality control of Yushu tablets.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drugs, Chinese Herbal ; analysis ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Tandem Mass Spectrometry

Objective To study the expression level of thyroid insulin-like growth factor-Ⅰ (IGF-Ⅰ) in iodine deficiency and excess mice and the effect of thyroid gland IGF-Ⅰ in the thyroid morphological change. Methods Forty-eight Balb/c mice were chosen as studied objects,weighing about 16 g. They were divided into three groups: low iodine(LI,iodine content of 50 μg/kg in feed,drinking deironized water) group,normoi(NI,iodine content of 300 μg/kg in feed,drinking deironized water) group and high(HI,iodine content of 300 μg/kg in feed,iodoine of content 14 700 μg/kg in drinking) group,16 mice in each group. Mice were put to death after 12 weeks and taken out of their thyroid gland. The body weight,absolute and relative weights of thyroid gland were measured and the morphological change of thyroid gland were observed under microscope. The expression levels of thyroid gland IGF-Ⅰ mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively. Results There were statistical significances between groups of thyroid absolute and relative weights(F = 315.881,405.921,all P < 0.01). LI group [(10.71±4.03) mg,(44.98±15.39)mg/100 g body weight]and HI group [(3.42±1.17)mg,(13.50± 3.89)mg/100 g body weight]had heavier thyroid absolute and relative weights than NI group[(2.11±0.53)mg,(8.35±1.98)mg/100 g body weight,all P < 0.01]. Under microscopy,the thyroid follicle capacity grew down and the follicle quantity grew up in LI group,the epithelium was stylolitic,the colloid diminished or absence in follicular cavity,while HI group presented colloid accumulation without follicular hyperplasia. The expression level of thyroid gland IGF-Ⅰ mRNA in LI group(1.03±0.32) was more than that in NI(0.65±0.19) and HI(0.59± 0.20) groups(F= 7.518,P< 0.01). In contrast to NI,there was no difference in the expression level of thyroid gland IGF-Ⅰ mRNA in HI group(P > 0.05). The brownish particles of LI group were more than NI and HI groups in the thyroid follicle epithelium by immunohistochemistry,while HI group was less than NI group. Conclusions The mice of iodine deficiency presented follicular hyperplasia goiter,the mice of iodine excess presented colloid accumulative goiter. The change of IGF-Ⅰ mRNA and protein expression may participate morphologleal change,indicating autocrine IGF-Ⅰ of thyroid gland may play an important role in regulating goiter formation.

Cantharidin is a terpenoid compound secreted by Mylabrisphalerata Pallas.In a variety of cancer types,cantharidin has shown the effects of inhibiting cancer cell growth,proliferation and migration.The anticancer mechanism of cantharidin involves inducing cell cycle arrest and apoptosis,inhibiting autophagy,enhancing DNA damage,inhibiting DNA repair,and regulating various cell signaling pathways.This article reviews the mechanism and research progress of cantharidin and its derivatives in common cancer types,in order to identify novel targets of cancer treatment using cantharidin,and provide new direction for clinical research to improve anticancer therapy.

OBJECTIVEThis study was designed to examine the in vitro effects of fenvalerate on steroid production and steroidogenic enzymes mRNA expression level in rat granulosa cells.

METHODSUsing primary cultured rat granulosa cells (rGCs) as model, fenvalerate of various concentrations (0, 1, 5, 25, 125, 625 micromol/L) was added to the medium for 24 h. In some cases, optimal concentrations of 22(R)-hydroxycholesterol (25 micromol/L), Follicle stimulating hormone (FSH, 2 mg/L), or 8-Bromo-cAMP (1 mmol/L) were provided. Concentrations of 17 beta-estradiol(E2) and progesterone (P4) in the medium from the same culture wells were measured by RIA and the steroidogenic enzyme mRNA level was quantified by semi-quantitative RT-PCR.

RESULTSFenvalerate decreased both P4 and E2 production in a dose-dependent manner while it could significantly stimulate rGCs proliferation. This inhibition was stronger in the presence of FSH. Furthermore, it could not be reversed by 22(R)-hydroxycholesterol or 8-Bromo-cAMP. RT-PCR revealed that fenvalerate had no significant effect on 3 beta-HSD, but could increase the P450scc mRNA level. In addition, 17 beta-HSD mRNA level was dramatically reduced with the increase of fenvalerate dose after 24 h treatment.

CONCLUSIONFenvalerate inhibits both P4 and E2 production in rGCs. These results support the view that fenvalerate is considered as a kind of endocrine-disrupting chemicals. The mechanism of its disruption may involve the effects on steroidogenesis signaling cascades and/or steroidogenic enzyme's activity.

3-Hydroxysteroid Dehydrogenases ; analysis ; metabolism ; 8-Bromo Cyclic Adenosine Monophosphate ; pharmacology ; Animals ; Base Sequence ; Cells, Cultured ; Dose-Response Relationship, Drug ; Estradiol ; analysis ; metabolism ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Hydroxycholesterols ; pharmacology ; Nitriles ; pharmacology ; Progesterone ; analysis ; metabolism ; Pyrethrins ; pharmacology ; RNA, Messenger ; analysis ; metabolism ; Rats ; Steroids ; metabolism

OBJECTIVETo observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α) in IL-1β induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages.

METHODSNaringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1β-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1β-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1β and TNF-α in IL-1β-induced CDs was detected by RT-PCR.

RESULTSThe appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1β on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α in IL-1β induced CDs (P < 0.05), which was approximate to the level of the normal group.

CONCLUSIONSNaringin could not only promote the proliferation of CDs, but also protect IL-1β-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1β and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Animals ; Blotting, Western ; Cartilage, Articular ; Caveolins ; Chondrocytes ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Interleukin-1beta ; metabolism ; Polypodiaceae ; Rabbits ; Rhizome ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Apoptosis ; physiology ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Carrier Proteins ; biosynthesis ; genetics ; DNA-Binding Proteins ; metabolism ; Gene Targeting ; Humans ; Liver Neoplasms ; enzymology ; pathology ; RNA Interference ; Telomerase ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo investigate the improvement of the Nuss procedure with the classification of the pectus excavatum by symmetry and extend the indication of the operation for a better outcome.

METHODSThe clinic data of 403 patients who underwent repair of pectus excavatum by Nuss technique and its modifications from July 2002 to September 2007 were reviewed retrospectively. There were 299 male patients and 104 female patients. The age ranged from 2 years and seven months old to 32 years old, with a mean of (8.0 +/- 5.1) years old. The entire group of the CT index ranged from 3.25 to 51.20, with a mean of (5.0 +/- 3.0). According to the morphology of the pectus and practically the bar shaping, Park's classification was simplified to symmetric type, eccentric type, and unbalanced type. The patients of symmetric type were all received original Nuss procedure, and the other two types underwent the procedure using characteristic bar shaping and technical modification.

RESULTSAmong the 403 patients, 257 patients (63.8%) were symmetric pectus excavatum, and 48 patients (11.9%) were eccentric, other 98 patients (24.3%) were unbalanced type. All the patients underwent the procedure successfully. The total time of the procedure ranged from 30 to 165 min, with a mean of (45.7 +/- 12.6) min. Volume of blood loss during the operation ranged from 1 to 80 ml, with a mean of (4.8 +/- 6.2) ml. The days of hospitalization ranged from 4 to 12 d, with a mean of (7.1 +/- 1.0) d. The duration of following up ranged from 1 month to 5 years, and the results of the repair were excellent in 391 patients (97.0%). Five patients received a further operation, including 1 patient of recurrent pectus excavatum caused by bar displacement, 4 patients of protruding side way. There were 23 patients of complications for an overall complication rate of 5.7%. And the other 18 patients of complications included 2 patients of pericardium perforation, 2 patients of bar displacements caused intercostal dilaceration, 2 patients of diaphragmatic muscle injuries, 1 patient of hemothorax, 8 patients of pneumothorax, 2 patients of intermittent pains for 2 months, 1 patient of scoliosis caused by persistent pain.

CONCLUSIONTo classify the pectus excavatum with symmetry and to choose different ways of procedure can extend the indication of Nuss procedure and receive a better outcome.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Funnel Chest ; surgery ; Humans ; Male ; Minimally Invasive Surgical Procedures ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo study the phenotypes and genotypes of a protein C (PC) deficiency pedigree.

METHODSImmunoassay (ELISA) was used for PC antigen and activated PC (APC) detection, PCR for amplification of the fragment of protein C gene exon II to exon IX, single-strand conformation polymorphism (SSCP) for difference of denatured cDNA and DNA sequencing for gene mutation.

RESULTSFour members in the pedigree were found to be PC antigen levels between 34.3% - 67.8% and PC activity between 22% - 49% which are lower in comparison with normal references (80% - 120% and 70% - 130%, respectively). A G-to-A mutation in exon VII of the protein C gene at 6 219 position was identified in 9 members. This mutation resulted in the substitution of Arg for Gln at 169 amino acid.

CONCLUSIONThe proband is of heterozygosity. The G6219 A mutation in exon VII of the protein C gene leads to the substitution of Arg 169 Gln. This mutation is reported for the first time in China.

Adult ; DNA Mutational Analysis ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Protein C ; genetics ; metabolism ; Protein C Deficiency ; congenital ; genetics

OBJECTIVETo study the chemical constituents from Lamium maculatum L. var Kansuense.

METHODThe chemical constituents were isolated and repeatedly purified by silica gel column chromatography and the structures were elucidated by the NMR spectra and physico-chemical properties.

RESULTTen compounds were obtained and they were identified as D-mannitol, beta-sitosterol, stigmasterol, rutin, 3'-methylquercetin-3-O-rutinoside, n-butyl-beta-D-fructopyranoside, daucosterol, acteoside, 20-hydroxyecdysone, allantoin.

CONCLUSIONAll the compounds were obtained from L. maculatum L. var Kansuense for the first time.

Allantoin ; chemistry ; isolation & purification ; Ecdysterone ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Mannitol ; chemistry ; isolation & purification ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rutin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Stigmasterol ; chemistry ; isolation & purification

BACKGROUND: The effect of extracellular matrix on stem cells is the focus of tissue engineering. However, there are few reports about the synthesis and secretion of extracellular matrix as well as its effects on cells. OBJECTIVE: To isolate, culture and identify rabbit bone marrow mesenchymal stem cells (BMSCs), and to explore the changes of extracellular matrix and whole structure under the intervention of ascorbic acid. METHODS: Rabbit BMSCs were isolated by differential adherent method of the bone marrow, and the expression of CD44, CD45 and CD31 was identified by flow cytometry. The BMSCs were cultured in the culture medium containing 20 mg/L ascorbic acid. Then the cell morphology, gross structure, ultrastructure, and histological changes of BMSCs were observed. The expression of extracellular matrix related genes was detected by RT-PCR. RESULTS AND CONCLUSION: Over 95% passage 2 BMSCs could express CD44, but the expression levels of CD45 and CD31 were extremely low. Intervention with ascorbic acid enhanced the proliferation of BMMSCs with unclear cell boundaries. A cell-sheet structure formed at 10-14 days after intervention. Hematoxylin-eosin staining results showed a layered cell arrangement, and Masson staining findings showed a large amount of extracellular matrix composition. Abundant endoplasmic reticula and vesicle-like structure were observed under the transmission electron microscope. RT-PCR findings showed that ascorbic acid significantly increased the expression of fibronectin mRNA in the BMSCs (P < 0.05), but slightly increased the mRNA expression of collagen type I. All these findings indicate that ascorbic acid not only increases the proliferation and transformation of rabbit BMSCs, but also promotes the synthesis and secretion of extracellular matrix, which has great potential in tissue engineering applications.