AIM: To research the processed products of Rhizoma Atractylodis Macrocephalae at the level of sesquiterpene lactones from it. METHODS: Sesquiterpene lactones were separated by the silica gel column and layer preparation, and the active substances were determined by HPLC. RESULTS: Atractylone、atractylolide Ⅰ, atractylolide Ⅲ, and biepiasterolid were separated from Rhizoma Atractylodis Macrocephalae. Atractylone was (0.535 8)%, atractylolide Ⅰ was (0.044 0)% and actractylolide Ⅲ was (0.081 4)% in Atractylodis stir-fried with wheat bran determined by HPLC. CONCLUSION: The method has good accuracy and repeatability and it can be used for the quality control of Rhizoma Atractylodis Macrocephalae.

An α-amylase inhibitor (α-AI) was isolated from white kidney beans (Phaseolus vulgaris. L) by ethanol fractional precipitation, ion exchange chromatography and gel filtration column chromatography. It was a homogeneity glycoprotein demonstrated by SDS-PAGE and gel filtration on CL-6B. The glycoprotein contained 88.2% protein and was rich in aspartic acid, glutamic acid, leucine, threonine and serine. The carbohydrate moiety was consisted of Man, Glc, Gal and Xyl in a mole ratio of 2.42∶1.50∶1.52∶1.00. The glycan and the core protein backbone was connected by O-linkage as determined by β-elimination reaction. The continuous oral administration of the α-AI (150 mg·kg-1·d-1 ) for 7 days can lower fasting blood glucose and 300 mg·kg-1 ·d-1 α-AI for 7 days can improve the sugar tolerance on alloxan-dependent diabetic model rats. The result showed the α-AI obtained from white kidney beans had good hypoglycemic effect on alloxan induced diabetic rats and may have high potential pharmaceutical value as a regulative digestive-starch degradation in patients suffering from diabetes.

Adult ; Aged ; Aged, 80 and over ; Humans ; Middle Aged ; Pneumoconiosis ; diagnostic imaging ; Radiography, Thoracic ; methods ; Tomography, X-Ray Computed ; X-Ray Film

Objective　To study the protocol that induces adult rat bone marrow stromal cells (MSCs) to express neuron-specific enolase (NSE) in vitro. Methods　MSCs were preinduced with β-mercaptoethanol for 24 h, and then induced for 5 h. The positive percentages of NSE protein expression were measured by immunocytochemistry SABC staining. Results　After the induction, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and processes which formed extensive networks. The positive percentage of NSE protein expression was (63.7±4.5)%. Conclusions　β-mercaptoethanol may induce adult rat MSCs to NSE-positive cells in vitro.

Objective To explore whether the transcription factor NF-κB takes part in baicalin-induced differentiation of bone marrow stromal cells (MSCs) into neurons. Methods MSCs from adult rats were induced by baicalin in the serum-free medium for 6 hs. The un-treated cells, as the control group, were only induced in the serum-free medium without baicalin. The expression of neuronal specific markers was evaluated by indirect immunofluorescence cytochemistry staining. The activity of NF-κB was measured by the presence of the NF-κB subunit RelA (p65) translocated into the nucleus in the same way. Results After the induction by baicalin, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and extensive networks of processes, and the expression of neuron-specific markers was detectable 6 ds after the induction. Neuronal specific marker proteins did not express in the control group. Six days after the induction, the P65 positive rate in the cytoplasm of the control group decreased to (18.4±3.0)%, while in the baicalin group, the P65 positive rate in the cytoplasm was (84.8±3.0)%. Conclusions Baicalin may inhibit the activation of NF-κB, which may act in the differentiation of MSCs into neurons.

Objective To explore the optimal HbAlc diagnostic cutpoint in different glucose tolerance populations in the plateau region.Methods (1) 472 diabetes mellitus (DM) patients and highrisk groups accepting diabetes screening in the First Affiliated Hospital of Kunming Medical College (217 males and 255 females,≥20 years old,median age 54 years old) were collected,oral glucose tolerance test (OGTT) and HbAlc were tested.(2) the research subjects were divided into normal glucose adjustment group (NGT),Impaired fasting glucose group (IFG) and (or) Impaired glucose tolerance IGT group and diabetes mellitus (DM) group.The receiver-operating characteristic curve (ROC) was explored to determine the optimal HbA1c diagnostic cut point for IFG,IGT and DM status respectively.Results The average HbA1 c values of NGT,IFG and (or) IGT,DM groups were (6.06 ± 0.11) ％,(6.63 ± 0.11) ％,(8.70 ± 2.08)％ respectively,for IFG and IGT groups,the optimal HbA1c diagnostic cut points were 6.7％ and 6.6％,respectively; If use either FBG or 2 h PG to diagnose DM,the corresponding optimal HbA1 c diagnostic cut point was 7.1％ ; If use anyone of FBG or 2hPG to diagnose DM,the corresponding optimal HbA1c diagnostic cut point was 7.0％ ; If both FBG and 2hPG were used to diagnose DM,the corresponding optimal HbA1 c diagnostic cut point was 7.1％.Conclusion Preliminarily confirm the optimal HbA1c diagnostic cut point in different glucose tolerance populations in the plateau region of Kunming,and provide the evidence for further clinical application of HbA1c.

We designed and synthesized eighteen lycorine derivatives with five different structural types, and evaluated their antiviral activities on a HCoV-OC43-infected H460 cell model. Structure-activity relationships suggested that the introduction of appropriate substituents on the 6N atom of lycorine was beneficial to activity. Compound 6a gave a good activity with the half effective concentration (EC₅₀) and selectivity index (SI) values of 2.36 μmol·L^-1 and 16.52, respectively. Surface plasmon resonance (SPR) result indicated that 6a might target the non-structural protein 12 (NSP12) subunit in RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with the dissociation constant (K_D) value of 1.36 μmol·L^-1. Molecular docking indicated that 6a might act on nidovirus RdRp-associated nucleotidyltransferase (NiRAN) catalytic center of NSP12, distinct from the mechanism of nucleoside-like drugs such as remdesivir. This study provides scientific data for the development of lycorine derivatives into a new class of anti-SARS-CoV-2 small molecule inhibitors.

OBJECTIVETo explore the experimental conditions for H2O2 to injure astrocytes and the effect of baicalin in protecting neurons and astrocytes from oxidative stress injury.

METHODSNeurons and astrocytes from forebrain of rats were cultured in vitro and treated with H2O2, baicalin and combination of the two, respectively. The cell viability or survival rate was determined using MTT.

RESULTSEffects of H2O2 in different concentrations on survival rate of astrocytes showed significant difference (F = 28.569, P < 0.01) in a dose-dependent manner. Degrees of H2O2 injury, with the same concentration of H2O2, on cells with different seeding density were also significantly different (F = 5.439, P < 0.01), and dose-dependently. Baicalin didn't influence the survival rate of neurons and astrocytes when the concentration was within 2.5-40 mumol/L (for neurons, F = 0.49, P > 0.05; for astrocytes, F = 1.001, P > 0.05), but baicalin showed significant antagonism to the injury of oxidative stress (for neurons, F = 24.384, P < 0.01; for astrocytes, F = 5.000, P < 0.01). The higher the concentration of bainalin, the higher the cell survival rate.

CONCLUSIONA model of astrocytes oxidative injury induced by H2O2 is established. Baicalin shows no toxicity on neurons and astrocytes when the concentration is within 2.5-40 mumol/L, but could antagonize the H2O2 caused oxidative injury on cells in a dose-dependent manner.

Animals ; Animals, Newborn ; Astrocytes ; pathology ; Cells, Cultured ; Flavonoids ; pharmacology ; Hydrogen Peroxide ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

Objective To explore the protocol that induced marrow mesenchymal stem cells (MSCs) differentiating into islet-secreting cells in vitro and to provide new clues for the sources of islet transplantation.Methods Using a defined culture medium and technique for transdifferentiation, MSCs from adult SD rats were guided into specific insulin-secreting cells. The expressions of nestin and islet-specific hormones and proteins, such as insulin, glucagon, somatostatin and pancreatic duodenal homeobox 1 (Pdx-1) were analyzed by indirect immunofluorescence cytochemistry staining before and after induction. The expressions of pancreatic islet cell differentiation-related transcripts, such as nestin, insulin 1, glucose transporter 2 (GLUT 2), Isl-1, Pdx-1, Pax-4 and Pax-6 were detected by reverse transcription-PCR (RT-PCR). In addition, the quantity of insulin secretion was examined using radioimmunoassay. Results Five hours after induction, (44.6±7.3)% of differentiated MSCs expressed nestin and it increased to (61.8±8.4)% 24 hs after induction, but the expression of nestin almost disappeared at day 14. In the meantime, islet-like cellular clusters appeared after day 14 and became more apparent by day 28. Differentiated cells were found to be immunoreactive to insulin, glucagon, somatostatin and Pdx-1, and expressed insulin 1, GLUT 2, GK, Isl-1, PDX-1, Pax-4, Pax-6 mRNA. In addition, the results of cumulative quantities of insulin of 24 hs and the stimulation index showed that differentiated cells were able to produce insulin at higher levels, and displayed glucose-dependent insulin release in vitro. Conclusions Adult rat MSCs can be differentiated into insulin-secreting cells in vitro. This approach might lead to widespread cell replacement therapy for Type 1 diabetes.

Objective To study the immune regulatory effect of natural killer cell(NKT) in the early stage of murine liver injury induced by type 5 adenovirus(Ad5). Methods Animal models were con-structed by injected C57BL/6 mice with 1.5×109-3×109 PFU Ad5 into the tail vein. Liver injury of mouse at day 0, 1, 2, 3, 4, 5 after infection was determined by HE staining and serum ALT/AST(alanine amin-otransferase/aspartate aminotransferase) level. Flow cytometry analysis was used to measure the proportion of lymphocytes, expression of Fas/FasL on the surface of NKT cells and level of IL-4, IFN-γ/in NKT cell plasma in the infected mouse liver. RT-PCR was applied to semi-quantify the chemokines and their receptors mRNA in infected mouse liver. Results NKT cells of mouse increased significantly at day 1 after infected with high titer Ads(3×109 PFU), expression of FasL on NKT cell and plasma IL-4, IFN-γ/level in NKT cells were also up-regulated, hence the obviously infiltration of lymphocytes in routine liver. Comparing with high titer Ads infection, low titer Ads infection (1.5×109 PFU) lead to little change of NKT cell proper-tion, and fewer infiltration of lymphocytes in murine liver. Hepatic chemokine RANTES, 1P-10, and MIP-1β mRNA expression in C57BL/6 was up-regulated 2 d after intravenous administration of 3×109 PFU Ad5. Corresponding chemokine receptor CCR5, CCR1, CXCR3 mRNA expression was up-regulated 3 d after in-fection. Conclusion NKT cells play an important role in lymphocytes recruitment into the liver of mouse in-fected with AdS, which may relate to up-regulatio of the plasma IL-4, IFN-γ level and expression of FasL of NKT cells, therefore facilitating the production of chemokines, e.g. IP-10 and Mig.