Animals ; Disease Models, Animal ; Hypoglycemia ; chemically induced ; metabolism ; Hypothalamus ; drug effects ; metabolism ; Insulin ; pharmacology ; Intracellular Signaling Peptides and Proteins ; metabolism ; Male ; Neuropeptides ; metabolism ; Orexins ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore effective pretreated methods for hydrosalpinx before frozen embryo transfer (FET).

METHODSA randomized controlled study was performed on 229 FET cycles of hydrosalpinx patients. They were assigned to two groups by random digit table, Group A (94 cases), Group B (89 cases), and Group C (46 cases). Patients in Group A received transvaginal aspiration of hydrosalpinx combined with auricular point sticking. Those in Group B received transvaginal aspiration of hydrosalpinx group. Those in Group C received no transvaginal aspiration of hydrosalpinx. Pregnancy outcomes of FET, endometrial and subendometrial blood flow distribution on the embryo transfer day were compared among the three groups.

RESULTSThere was no statistical difference in the endometrial thickness on FET day, the number of transfer embryos, the number of transferred good quality embryos among the three groups (P > 0.05). The clinical pregnancy rate and the embryo implantation rate were significantly higher in Group A than in Group C (P < 0.05), and the clinical pregnancy rate was significantly higher in Group A than in Group B (P < 0.05). The early abortion rate and the transfer cycle cancel rate were significantly lower in Group A than in Group C (P < 0.05). Type A endometrial and subendometrial blood flow distribution was dominant in Group A, which was significantly higher in Group A than the rest two groups (P < 0.05). Type A distribution rate was also significantly higher in Group B than in Group C (P < 0.05).

CONCLUSIONTransvaginal aspiration of hydrosalpinx combined with auricular point sticking before FET could improve the endometrial receptivity and improve outcomes of IVF.

Embryo Implantation ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Humans ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate

Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

OBJECTIVETo study the effects of Danggui Buxue Decoction on the hematopoietic reconstruction of mice transplanted by muscle satellite cells ( MSCs).

METHODSMSCs were procured from newly born male mice of the homologous series. The female Kunming receptor mice irradiated with 8Gy137Cs-gamma ray were randomly divided into six groups, i. e., the blank control group, the transplanted MSC group, four groups intervened by different doses of Danggui Buxue Decoction after transplanted MSC (gastrogavage by one, three, five, and ten times of clinically equivalent dose for seven days, as the DGBX 1 group, the DGBX 2 group, the DGBX 3 group, and the DGBX 4 group). After transplantation changes of splenic colony forming unit spleen (CFU-S), white blood cells (WBC), hemoglobin (Hb), and platelet (PLT) in the peripheral blood were observed in pos-transplanted 1-, 2-, and 3-week receptor mice. The 3-week survival rate was calculated. The source of hematopoietic reconstruction was identified using PCR.

RESULTSThe desmin protein was positive in the cultured MSCs. WBC of each transplanted MSC group obviously increased at the 2nd week (P<0.05). Hb obviously increased in the DGBX 2 group, the DGBX 3 group, and the DGBX 4 group (P<0.05). WBC and Hb obviously increased in the DGBX 3 group and the DGBX 4 group when compared with the transplanted MSC group at the 3rd week (P<0.05). The recovery of PLT was significant in the DGBX 4 group (P<0.05). Compared with the blank control group at the 2nd week, CFU-S obviously increased in the DGBX 3 group and the DGBX 4 group (P<0.05). PCR results of Y chromosome in the survived transplant mice indicated that the hematopoietic cells of reconstruction female receptor mice were originated from male donors.

CONCLUSIONThe hematopoiesis of mice transplanted by muscle satellite cells could be constructed after intervention of Danggui Buxue Decoction.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hematopoiesis ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred Strains ; Satellite Cells, Skeletal Muscle ; drug effects ; transplantation

OBJECTIVETo investigate the chemical constituents of dried whole plants of Artemisia annua.

METHODThe chemical constituents were isolated by repeated silica gel chromatography, medium pressure column chromatography, and semi-preparative HPLC, and their structures were elucidated by spectroscopic analyses and comparison of NMR data with those reported in literature.

RESULT15 compounds were isolated and identified to be 5-O-[(E)-Caffeoyl] quinic acid(l), 1,3-di-O-caffeoylquinic acid(2), 4 5-di-O-caffeoylquinic acid(3), 3, 5-di-O-caffeoylquinic acid (4), 3, 4-di-O-caffeoylquinic acid (5), methyl-3,4-di-O-caffeoylquinic acid(6), methyl-3,5-di-O-caffeoylquinic acid(7), 3,6'-O-diferuloylsucrose(8), 5'-β-D-glucopyranosyloxyjasmonic acid(9), Scopoletin(10), scoparone (11), 4-O-β-D-glucopyranosyl-2-hydroxyl-6-methoxyacetophenone (12), chrysosplenol D (13), casticin (14), chrysosplenetin(15).

CONCLUSIONCompounds 2, 6, 8 and 9 are obtained from the Artemisia genus for the first time. Compounds 7 and 15 are obtained from this plant for the first time.

Artemisia annua ; chemistry ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Medicine, Chinese Traditional ; Plants, Medicinal ; Quinic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Silica Gel

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

OBJECTIVETo purify caffeic acid tetramer (CAT) with macroporous resin on the basis of its fundamental physicochemical stability research.

METHODThe changes of CAT content were compared by HPLC method before and after the purification process, or while other conditions were altered.

RESULTLK001 was the best one among 7 kinds of macroporous resin in regard of purifying ability. The optimum absorbing technology was the solution concentration at 10 g x L(-1), pH at 4.5, and the flow rate at 3 BV x h(-1). The best eluting technology was 45% ethanol as eluting agent, pH at 5.0, eluting volume at 50 mL after applying super-purified water and 20% ethanol. The yield of product was 3. 6 percent, and the active compound CAT was 58 percent in the product.

CONCLUSIONMacroporous resin LK001 is effective in enriching CAT from the crude extracts, thus this method of purification is advisable.

Absorption ; Boraginaceae ; chemistry ; Caffeic Acids ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Hydrogen-Ion Concentration ; Light ; Oxygen ; chemistry ; Porosity ; Resins, Plant ; chemistry ; Temperature ; Water ; chemistry

Macrophage migration inhibitory factor (MIF) is a widely expressed multipotent cytokine that participates and plays an important role in various inflammatory and immune diseases‚ and is a biomarker or therapeutic target of many diseases. MIF is highly conserved in phylogeny and there are specific binding sites of various transcription factors in its promoter region‚ which can regulate the expression of MIF. MIF functions both inside and outside cells‚ and MIF is constitutively expressed. Therefore‚ it is of great significance to study the related factors that regulate MIF gene expression and stimulate MIF secretion. This article summarizes and classifies the related factors affecting MIF gene expression by briefly describing the binding sites on the MIF gene and MIF promoter. According to the way of binding with the MIF gene‚ it can be divided into:(1) binding to specific sites of MIF gene promoters to change transcription activity; (2) binding to MIF CATT5-8 microsatellite repeats to change highly expressed MIF alleles (3) non-coding RNAs regulating MIF expression; (4) related factors affecting MIF secretion. By reviewing the four types of related factors that regulate MIF gene expression‚ we will understand the regulatory mechanism and influencing factors of MIF gene expression‚ in order to provide a theoretical basis for its treatment of related diseases.

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism

OBJECTIVETo observe the effects of angelica polysaccharides (APS) on the proliferation of mouse skeletal muscle satellite cells (MSCs) and c-kit expression in different in vitro hematopoietic microenvironments.

METHODSMSCs were primarily cultured. The desmin protein was examined by immunohistochemical assay five days later. The MSCs were randomly divided into 8 groups, i. e., the control group, the supernatant from cultured bone marrow stroma cells group, 100, 200, 300 microg/mL APS added in the DMEM/F12 medium experimental groups, and 100, 200, 300 microg/mL APS intervened medium groups. The effects of the proliferation activities of MSCs were detected using MTT method. The c-kit protein of the MSCs was stained by immunohistochemistry.

RESULTSThe desmin protein was positive in the isolated cultured MSCs. Results of MTT method showed the proliferation of MSCs in APS intervened medium groups was significant. The strong positive c-kit immunoreactivity existed in APS intervened medium groups. The strong positive c-kit immunoreactivity was present in the cytoplasmic of the MSCs in the DMEM/F12 medium experimental groups and the APS intervened medium groups.

CONCLUSIONSThe APS intervened MSC medium could effectively change the growth properties of MSCs, obviously promote the proliferation of MSCs and c-kit expression. The c-kit protein might play some regulative roles in the proliferation of the MSCs.

Angelica ; Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Male ; Mice ; Mice, Inbred Strains ; Polysaccharides ; pharmacology ; Proto-Oncogene Proteins c-kit ; metabolism ; Satellite Cells, Skeletal Muscle ; drug effects ; metabolism