Objective To explore the effect of different processes on the contents of active ingredients in Rhizoma Corydalis. Methods Processes of vinegar-boiling, vinegar-frying and alcohol-frying were used to fresh corydalis, boiled corydalis and purchased corydalis. Then the method of extraction by ultrasound with water and by reflux with methanol were used to extract the sample. The contents of Protopine, Tetrahydropalmatine and Dehydrocorydaline in corydalis was determined by RP-HPLC. Agilent TC-C18 (25 mm?4.6 mm, 5 ?m) column was used at 35 ℃, with acetonitrile-0.1% phosphonic acid (pH 5.3 with triethylamine) (28∶72) as mobile phase. The flow rate was 1 mL/min and detection wavelength was set at 280 nm. Results In the sample extracted by water, the contents of active ingredients in processed products were higher than in corydalis. And in the sample extracted by methanol, it also has the tendency that the contents of active ingredients in processed products were higher than in corydalis. The content of Dehydrocorydaline in fresh corydalis which was processed by vinegar-boiling, vinegar-frying and alcohol- frying was high. The content of Tetrahydropalmatine in purchased corydalis which was processed by water-boiling at production place was high. Conclusion The method of determining active ingredients in Rhizoma Corydalis was established. It was convenient and the result was accurate. The process technology that fresh corydalis was boiled by water and then processed or fresh corydalis was processed in production place can be used.

Isoliquiritigenin (ISL) is an active chalcone compound isolated from licorice. It possesses anti-inflammatory and anti-oxidative activities. In our previous study, we uncovered a great potential of ISL in treatment of type 2 diabetes mellitus (T2DM). Therefore, this study aims to reveal the mechanism underlying the alleviatory effects of ISL on T2DM-induced glycolipid metabolism disorder. High-fat-high-sugar diet (HFD) combined with intraperitoneal injection of streptozotocin (STZ) were used to establish T2DM mice model. All animal experiments were carried out with approval of the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments, and sodium palmitate (SP) was applied to establish insulin resistance (IR) model cells. The effects of ISL on body weight, fasting blood glucose levels, and pathological changes in the livers of mice were examined. Enzyme-linked immune sorbent assay (ELISA) and real-time quantitative PCR (RT-qPCR) were applied to detect the regulatory effects of ISL on key targets involved in glucolipid metabolism. Additionally, molecular docking and analytical dynamics simulation methods were used to analyze the interaction between ISL and key target protein. The results indicate that ISL significantly downregulates the transcriptional levels and inhibits the activities of key enzymes involved in gluconeogenesis, including pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), and fructose-1, 6-bisphosphatase (FBP). It also downregulates the transcriptional and protein levels of hepatocyte nuclear factor 4α (HNF4α) and cAMP response element binding protein (CREB), the two transcriptional factors involved in gluconeogenesis. Thus, ISL inhibits hepatic gluconeogenesis in T2DM mice. In addition, ISL reduces total cholesterol (TC) and triglyceride (TG) levels in the livers of T2DM mice. Moreover, ISL downregulates the mRNA levels of lipogenesis genes and upregulates those of genes involved in fatty acid oxidation, lipid uptake, and lipid export. In conclusion, ISL suppresses hepatic gluconeogenesis, promotes lipolysis, and restrains lipogenesis in T2DM mice, thereby improving the abnormal glycolipid metabolism caused by T2DM.

Isoliquiritigenin (ISL) is a flavonoid compound isolated from licorice. It possesses excellent antioxidant and anti-diabetic activities. This study aims to investigate the molecular mechanism underlying the alleviatory effect of ISL on energy metabolism imbalance caused by type 2 diabetes mellitus (T2DM). 8-week-old male C57BL/6J mice were used in in vivo experiments. The high-fat-high-glucose diet combined with intraperitoneal injection of streptozotocin was applied to establish T2DM animal model. All animal experiments were performed in accordance with the Institutional Guidelines of Laboratory Animal Administration issued by the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments. Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to examine the protein and mRNA levels of mitochondrial function-related targets. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) in HepG2 cells were measured by the flow cytometry. Additionally, the molecular docking of ISL and key target proteins was analyzed. It was found that ISL significantly inhibited the activity of mitochondrial respiratory chain complex I and increased the protein levels of uncoupling protein 2 (UCP2) in the livers of mice and HepG2 cells. It also obviously decreased the ROS levels and increased the MMP levels in cultured HepG2 cells. In addition, ISL promoted mitochondrial biogenesis by activating proliferator-activated receptor gamma co-activator 1α (PGC-1α) and enhanced mitophagy by upregulating Parkin. It also improved mitochondrial fusion by increasing the mRNA and protein levels of mitofusin 2 (MFN2). In conclusion, ISL alleviates energy metabolism imbalance caused by T2DM through suppression of excessive mitochondrial oxidative phosphorylation and promotion of mitochondrial biogenesis, mitophagy, and fusion.

Background The incidence and progression of glaucomatous optic neuropathy is related to a disturbance of the retrobulbar hemodynamics.Compound anisodine is clinically applied for the treatment of ischemic ocular diseases.Objective To evaluate the effects of compound anisodine injection on the ocular blood flow of glaucoma patients.Methods Twenty-one patients with primary glaucoma were divided into the treatment group and the control group.The eyes of each patient in the treatment group were selected further into the treatment eye group (11 eyes with greater mean deviation [MD]) or the opposite eye group(11 eyes with lesser MD).One of the eyes of each patient in the control group with MD value were selected as control eyes (10 eyes).The treatment eye group received compound anisodine on the para-superficial temporal artery via subcutaneous injection once a day for 2 treatment periods (each period equals 14 days,with 7 days intermittent between periods,totally 35 days) in addition to routine treatment.The retrobulbar blood flow,optic disc data,refraction error,visual field and intraocular pressure were measured in 3 time points:Before treatment period (baseline test),one day after treatment period (the 1st postreatment test) and 35 days after treatment period (the 2nd posttreatment test).Results Compared with the control group,the peak systolic velocity (PSV) and end diastolic velocity (EDV) of short posterior ciliary artery (SPCA) of the treatment eye group were relatively increased significantly in the 1st posttreatment test (P =0.017,0.028),the PSV of SPCA of the opposite eye group was relatively increased significantly in the 1 st posttreatment test (P =0.049),the EDV of central retinal artery (CRA) of the opposite eye group was relatively increased significantly in the 2nd posttreatment test (P =0.035).In contrast to the treatment eye group,the inferior quadrant RNFL thickness of the optic disc decreased significantly in the 2nd posttreatment test in the control group (P =0.009),the 6 o'clock RNFL thicknesses of the optic disc decreased significantly in the 1 st posttreatment test in the control group (P =0.014),and the 6 o'clock and 7 o'clock RNFL thicknesses of the optic disc decreased significantly in the 2nd posttreatment test in the control group (P =0.029,0.011).Conclusions The application of compound anisodine for the treatment of primary glaucoma relatively increases the PSV and EDV of SPCA.

Background Retinal retinoic acid (RA) plays an important role in the formation of the lensinduced myopia.However,it is not clear how RA transfer the myopic signal to choroid throughout the retinal pigment epithelium (RPE) barrier.Objective The aim of this study was to investigate the effect of all-trans retinoic acid (atRA) on the barrier of RPE in lens-induced myopic eye of guinea pig.Methods Thirty left eyes of 30 21-dayold clean guinea pigs were randomized into normal control group and the model group.The models of out of focus were induced by covering of-6.00 D concave lens on the left eyes for 15 days.Radius of corneal curvature was measured using corneal curvimeter,and diopeter of the guinea pig was examined by mydriatic optometry.The length of ocular axis was detected by A-sonography.The animals were sacrificed and the retinas of the left eyes were isolated for the culture and passage of RPE cells.The third generation of cells were incubated Millcell-PET microporous film,and atRA at the concentration of 1 × 10-6,1 × 10-7,1 × 10-8 and 1 × 10-9 mol/L was added to the micropore respectively for 12 hours,and the micropores with equal-solvent served as negative control group.Methyl thiazolyl tetrazolium (MTT)colorimetric method was used to detect the survival rate of the cells.Subsequently,the transepithelial electrical resistance (TER) of the monolayer cells was determined with CN10-EVOM2 resistance measuring meter.The vesicular transport change of RPE membrane in different groups was evaluated by FM1-43 fluorescence staining.Results The mean diopter was (-2.20±0.95) D in the models,and that in the controls was (+ 1.15 ±0.30) D,with a significant difference between them (t =14.57,P＜ 0.01).The axial length was (8.24 ± 0.09) mm in the models and it was significantly longer than (7.81±0.05) mm in the controls (t=17.20,P＜0.01).RPE cells grew well to form a monolayer in Millcell culture pool after one week.After 24 hours of the atRA treatment,the survival rate of RPE cells reduced gradually with the increase of atRA concentration with the highest rate in the 1 × 10-9 mol/L atRA group (93.3 ％) and followed by the 1 × 10-8 mol/L atRA group (88.2％).More than half of the cells dead in the 1 × 10-6mol/L and 1 × 10-7mol/L atRA groups (53.8％ and 47.1％).Significant differences in the TER value and fluorescence staining intensity of the cells were seen among the various groups (F =43.89,P =0.00 ; F =26.13,P =0.00),with the maximal values in the 1 × 10-8mol/L atRA group.The FM1-43 fluorescence located on the cellular membrane and cytoplasm.Conclusions AtRA can increase the functional state of tight junction and vesicular transport,which regulated the RPE cell barrier in the guinea pig.

The incidence of malignant melanoma, a highly fatal skin tumor, is on the rise worldwide. Melanomas are highly aggressive and have strong metastatic capability that leads to lethality. Recurrence occurs in patients with distant metastases, even with the latest treatments, and median survival is only a few months. At present, the prevention and treatment of melanoma includes surgical resection, chemotherapy, immunotherapy and targeted therapy. However, these strategies can lead to drug resistance and adverse effects. In recent years, an increasing number of studies have found that natural products have effective anti-melanoma activities, including inhibition of tumor growth, induction of cell apoptosis, inhibition of angiogenesis and metastasis and toxicity to tumor stem cells. In addition, several studies have reported that the combination of natural products and traditional anti-melanoma drugs can enhance the therapeutic efficacy. In this review we summarize the prevention and treatment of melanoma with natural products.

An Aspergillus sp. strain F3 was isolated and identified from the rhizosphere soil of mangrove plant, Rhizophora stylosa Griff in Dongzhai harbor mangrove forest conservation in China. In this study, the effects of media salinity and pH on the mycelial biomass and the ability of producing antibacterial metabo- lites from this isolate were carefully analyzed. Results showed that this isolate can grow well on the SDA medium with higher salinity (3%~9%) and higher pH (8~10). Under the modified culturing conditions, this isolate can secret the antibacterial and antitumor metabolites. The extracts of acetic ether were about 448 mg/L of the fermentation broth. The antibacterial activities of the acetic ether extract were analyzed with bacteria and fungus. Results showed this extract can suppress the growth of Staphylococcus aureus、S. epi-dermidis、Sarcina lutea、Bacillus subtilis and Escherichia coli with MIC of 31.3 ?g/mL, 31.3 ?g/mL, 7.8 ?g/mL, 7.8 ?g/mL and 125.0 ?g/mL, respectively. It can also suppress the growth of Candida albicans with MIC of 125.0 ?g/mL. Further studies uncovered the cytotoxicity of this extract against the tumor cells, such as ECV304, Lovo and HepG2 with IC50 of 3.45 ?g/mL, 4.88 ?g/mL and 14.31 ?g/mL respectively.

OBJECTIVETo explore the effect of α- galactosyleramide( α-GalCer ) on immune reconstitution under acute graft-versus-host disease (aGVHD).

METHODSBALB/c mice were transplanted wit hallogeneic C57BL/6 bone marrow cells and splenocytes (both 1×10(7))after receiving lethal total-body irradiation. α-GalCer (100 ug/kg) or vehicle (dimethylsulfoxide) was administered intraperitoneally immediately after transplantation. The effects of α-GalCer on immune reconstitution,proliferation of T cells and B cells, hematopoiesis,and thymic microenvironment were assessed.

RESULTSThe α-GalCer group exhibited higher percentages of CD3(+),CD4(+), CD8(+), B220(+), CD40(+), and CD86(+)cells compared with the vehicle group . The number of colony forming unit per 1000 CD34(+) cells in the α-GalCer group was higher than in the vehicle group ( P=0.0012).In vitro proliferation assays showed that the α-GalCer group had higher percentages of CD3(+), CD4(+), CD8(+),and B220(+) cells compared with the vehicle group. As for the results of in vivo proliferation assays, the numbers of CD3(+), CD4(+), CD8(+), and B220(+)cells were higher in the α-GalCer group than in the normal group ,especially the number of B220(+) cells ( P=0.007).Significant difference was not found in thymocyte count between the α-GalCer group and the vehicle group, nor in the percentages of CD3(+), CD4(+), and CD8(+) cells.

CONCLUSIONAdministration of α-GalCer after allogeneic bone marrow transplantation may promote immune reconstitution in the presence of aGVHD.

Animals ; B-Lymphocytes ; drug effects ; immunology ; Bone Marrow Transplantation ; immunology ; Female ; Galactosylceramides ; pharmacology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cells ; drug effects ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes ; drug effects ; immunology ; Transplantation, Homologous

Objective To investigate the effect of phosphoinositide-3-kinase inhibitor LY294002 on the differentiation of human embryonic stem cells (HESC) into more mature insulin-producing cells. Methods HESCs were induced to differentiate into insulin-producing cells through five stages. Nicotinamide and B27 (group B27), nicotinamide and LY294002 (group LY) were used to induce the nesting positive cells into mature insulin-producing cells. The morphological change of each stage was observed under microscope , and expressions of insulin, c-peptide, somatostatin and glucagon were identified by immunofluorescence staining. Results After 14 days in stage 5 , there was no significant difference in rate of insulin positive cells between group LY and group B27 (P﹥0.05), but rates of somatostatin and glucagon positive cells in group LY were lower than those in group B27(P﹤0.05). Furthermore, the co-stained rate of somatostatin and insulin in group LY was also lower than that in group B27 (P﹤0.05). Conclusion HESCs can be induced to differentiate into more mature insulin-producing cells by phosphoinositide-3-kinase inhibitor LY294002 in serum-free culture medium.

Objective To find out the iodine nutritional status and thyroid function of pregnant and lactating women in the urban areas of Wuwei City of Gansu Province,and to provide a scientific basis for iodine supplementation.Methods Blood and urine samples of pregnant (52 persons) and lactating women (59 persons) were collected in 2009.Urinary iodine content was measured with arsenic cerium catalytic spectrophotometric.Serum free three triiodothyronine(FT3),free thyroid hormone(FT4),triiodo thyronine(TT3),serum total thyroxine (TT4) and thyroid-stimulating hormone (TSH) were detected with direct chemiluminescence immunoassay.Results The medians urinary iodine of pregnant and lactating women were 274.68,217.88 μg/L.The rates of low urinary iodine (pregnant women below 150 μg/L,lactating women lower than 100 μg/L) were 9.62％ (5/52) and 6.78％ (4/59).Serum TT3,TT4 levels in pregnant women[(2.48 ± 0.59),(132.18 ± 33.36)nmoL/L] were higher than that in the lactating women[(2.16 ± 0.49),(108.79 ± 28.36)nmol/L,t =-3.123,-3.971,all P ＜ 0.05].Thyroid dysfunction incidence rates of pregnant and lactating women were 17.31％ (9/52) and 8.47％ (5/59).Thyroid dysfunction was mostly subclinical hypothyroidism.Conclusions Overall iodine nutrition of pregnant and lactating women is in good condition,some individuals have the trend of hypothyroidism.It is necessary to carry out routine monitoring of urinary iodine and thyroid function.