UPLC-Q-TOF-MS analysis of chemical constituents of classical prescription Yiguanjian standard decoction / 中国中药杂志

An UPLC-Q-TOF-MS method was employed to characterize and classify the chemical components of the standard decoction of Yiguanjian, a classical famous recipe. Chromatographic separation was performed on an Acquity HSS T3(2.1 mm ×100 mm, 1.8 μm) column with a mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile using gradient elution. The flow rate was 0.4 mL·min~(-1) and the column temperature was 40 ℃. Mass spectrometry was performed on electrospray ionization source(ESI) with positive and negative ion scanning modes. The potential compounds were identified by comparing the reference compounds, analyzing the mass spectrometry data and matching the published articles on Masslynx 4.1 software and SciFinder database. Finally, a total of 113 compounds, including 11 amino acids, 19 terpenoids, 13 phthalides, 11 steroidal saponins, 10 coumarins, 9 alkaloids, 7 flavonoids, 8 phenylethanoid glycosides, 8 organic acids and 17 other categories were identified. The established method systematically and accurately characterized the chemical components in Yiguanjian, which could provide experimental evidences for the subsequent studies on the pharmacodynamical material basis and quality control of Yiguanjian.

Physalin B reduces Aβ secretion through down-regulation of BACE1 expression by activating FoxO1 and inhibiting STAT3 phosphorylation / 中国天然药物

Physalin B (PB), one of the major active steroidal constituents of Solanaceae Physalis plants, has a wide variety of biological activities. We found that PB significantly down-regulated β-amyloid (Aβ) secretion in N2a/APPsw cells. However, the underlying mechanisms are not well understood. In the current study, we investigated the changes in key enzymes involved in β-amyloid precursor protein (APP) metabolism and other APP metabolites by treating N2a/APPsw cells with PB at different concentrations. The results indicated that PB reduced Aβ secretion, which was caused by down-regulation of β-secretase (BACE1) expression, as indicated at both the protein and mRNA levels. Further research revealed that PB regulated BACE1 expression by inducing the activation of forkhead box O1 (FoxO1) and inhibiting the phosphorylation of signal transducer and activator of transcription 3 (STAT3). In addition, the effect of PB on BACE1 expression and Aβ secretion was reversed by treatment with FoxO1 siRNA and STAT3 antagonist S3I-201. In conclusion, these data demonstrated that PB can effectively down-regulate the expression of BACE1 to reduce Aβsecretion by activating the expression of FoxO1 and inhibiting the phosphorylation of STAT3.

UPLC-Q-TOF-MS analysis on chemical constituents of classical prescription Huangqi Guizhi Wuwu Tang standard decoction / 中国中药杂志

UPLC-Q-TOF-MS technology was used to analyze the chemical constituents from classical prescription Huangqi Guizhi Wuwu Tang standard decoction. Acquity HSS T3 column(2.1 mm × 100 mm, 1.8 μm) was used as the chromatographic column, with 0.1% formic acid solution-0.1% formic acid acetonitrile as the mobile phase for gradient elution. The volume flow rate was 0.4 mL·min~(-1) and the column temperature was 40 ℃. Mass spectrometry data of Huangqi Guizhi Wuwu Tang standard decoction were collected in positive and negative ion modes. The chemical constituents from classical prescription Huangqi Guizhi Wuwu Tang standard decoction were analyzed and identified by Masslynx 4.1 software combined with SciFinder database, comparison with reference mate-rials, mass spectrometry data analysis and reference to relevant literature. A total of 110 compounds were analyzed and identified, including 33 flavonoids, 14 monoterpene glycosides, 8 triterpenoids, 8 gingerols, 17 phenylpropanoids, 12 organic acids, 7 amino acids and 11 other compounds. The results of this study provide an experimental basis for the further research on the substance basis and quality control of Huangqi Guizhi Wuwu Tang standard decoction.

Application of Photoshop in Measurement of Microsphere Diameter and Microstructure Drawing of Medicinal Materials / 中国实验方剂学杂志

<b>Objective::To explore the feasibility of Photoshop image processing software in the micro-spherical diameter measurement and identification of medicinal materials, in order to provide a new method for the measurement and mapping of traditional Chinese medicine. <b>Method::Photoshop (CS3 version and above) software was used to measure the diameter of 200 Notoginseng Radix et Rhizoma albumin microspheres in the same batch, digital microscopy and Photoshop image processing software was used to draw schematic diagrams of microstructures of Notoginseng Radix et Rhizoma medicinal powders and its four common pseudo-products and powders, namely Bletillae Radix, Alpiniae Officinarum Rhizoma, Curcumae Longae Rhizoma and <italic>Manihot esculenta,</italic> and make the identification table for classification and identification. <b>Result::The average diameter of the microspheres was (30.62±4.21)μm, and the diameter was mainly distributed in 20-40 μm. The average diameter of the microspheres was verified by laser particle size analyzer (30.18±4.67)μm. The differences between the two methods were not statistically significant. There were many starch granules in Notoginseng Radix et Rhizoma powder, with no calcium oxalate needle, oil cell, brown pigment and stone cell. Microscopic identification could be made for Notoginseng Radix et Rhizoma powder and 4 common counterfeits. <b>Conclusion::Photoshop image processing software measures the diameter of the microspheres with a high speed, high accuracy, simplicity, and low requirements for measuring instruments. It provides a new way to quickly measure the diameter of microspheres. Photoshop image processing software draws a schematic diagram of the microstructure, which is convenient and faster. The original morphology of the ground-reactive microstructure provides a new method for the microstructural drawing.

The effects of painful procedures during neonatal period on rats' hippocampal gene expression profiles in later life / 中国新生儿科杂志

Objective To study the effects of painful procedures during neonatal period on rats' hippocampal gene expression in later life.Method A total of 28 newborn male rats were randomly assigned into two equal groups.From day1 (P1) to day7(P7) after birth,rats in the pain group received four times of needle acupuncture at the foot everyday,simulating the infants' experience in the NICU.Rats in control group were touched at the same side of the foot with a cotton swab.Transcriptome sequencing of the hippocampus of the two groups were examined on day 8 (P8) and day 21 (P21).The gene expression profiles were established and the differentially expressed genes were screened for functional analysis.Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to confirm these differentially expressed genes (GABRB1,GRIN2A,IL1 RAPL1) related to pain-stimulated response or brain cognition,and one of the key genes was further verified using Western blotting.Result The sequencing results showed that there were only 6 differentially expressed genes in hippocampal tissues of rats on P8 in the two groups.Howerer,the number increased to 53 on P21 and 85.0％ of these genes were down-regulated (45/53).Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the differentially expressed genes were mainly expressed on the cell membrane,voltage-gated ion channels,synapses,neurotransmitter receptors,immune responses,etc.The qRT-PCR and Western-blot results of key genes were consistent with the transcriptome sequencing.Conclusion Pain stimuli at an early stage after birth may trigger differentially expression of voltage-gated ion channel proteins,neurotransmitter receptors,and some key genes such as GABRB1 on hippocampal synaptic cell membranes in rats.These phenomenon may provide initial explanation for the molecular mechanism of early pain stimuli on neonatal brain development.

Isolation and identification of exosomes derived from bone marrow mesenchymal stem cells / 基础医学与临床

Objective To develop a method of isolation and identification of exosomes of bone marrow mesenchymal stem cells.Methods The mesenchymal stem cells were cultured by whole bone marrow adherence method.A new reagent -exosomes extraction kit was used to isolate and collecte exosomes.The exosomes were identified by elec-tron microscopy,particle size detection,flow cytometry and Western blot.Results The expression of CD45 on the surface of the third generation bone marrow mesenchymal stem cells was negative,and CD73 and CD105 were posi-tive;exosomes derived from bone marrow mesenchymal stem cells were round or oval,the size is non-uniform,the diameter is 30~100 nm,have a complete membrane structure,and containing low-density substances;Particle size detection particle diameter of the main peak was 61.25 nm, in which the diameter of particles was about 20-200 nm accounted for 72.4%;exosome expressed CD63 and CD81;The expression of CD9 and CD63 from cell cul-ture supernatants was positive.Conclusions The exosomes can be collected in the medium of mesenchymal stem cells.The exosomes derived from bone marrow mesenchymal stem cells can be identified by electron microscopy, particle size detection,flow cytometry and Western blot.

Mechanism of MBL inhibiting the LPS-induced DC maturation / 中国实验血液学杂志

The study was aimed to investigate the mechanism of mannan-binding lectin (MBL) on bacterial lipopolysaccharide (LPS)-induced human peripheral blood monocyte-derived dendritic cell (DC) maturation. The monocytes were prepared from the peripheral blood of healthy adult volunteers. The immature dendritic cells (imDC) were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4. FACS was used to investigate the interaction of MBL with imDC and the impact of MBL on LPS binding to imDC. ELISA and Western blot was used to analyze the interaction of MBL with soluble TLR4 ectodomain protein (sTLR4); Western blot was used to detect LPS-induced NF-κB translocation in imDC. The results showed that MBL could directly bind to imDC in the presence of calcium. sTLR4 protein or LPS could competitively inhibit the binding of MBL to imDC. ELISA and Western blot showed that MBL could evidently bind to sTLR4 protein in a concentration-dependent manner. FACS showed that MBL could competitively inhibit the binding of LPS to imDC by binding to imDC directly. Western blot showed that MBL decreased LPS-induced NF-κB translocation in imDC. It is concluded that MBL may competitively inhibit the binding of LPS to imDC by binding to TLR4 expressed on imDC, resulted in inhibition of LPS-induced DC maturation, suggesting that MBL can regulate DC maturation through ligand-binding. This study provides the good foundation to clarify the mechanism of MBL inhibiting the LPS-induced DC maturation.

Effect of progesterone on expression of brain tissue damage factors and anti damage factors in neonatal rats with hypoxic-ischemic brain damage / 中华实用儿科临床杂志

Objective To study the effect of progesterone on tumor necrosis factor (TNF-α),interleukin-1β (IL-1β),nerve growth factor(NGF) and brain derived neurotrophic factor(BDNF) expression in the cortex and hippocampus tissue in newborn rats with hypoxic-ischemic brain damage (HIBD),and to discuss the protective molecular mechanism of progesterone on HIBD in neonatal rats.Methods Forty-eight 7-day-old neonatal rats were randomly divided into 3 groups:sham-operated group,hypoxic-ischemic group and pretreatment group.Rats in hypoxic-ischemic group and pretreatment group were subjected to left common carotid artery ligation,then they were exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in the closed container at 37 ℃ for up to 2.5 h to establish HIBD models.Progesterone was injected intraperitoneally into the rats in the pretreatment group 30 min before hypoxia,and solution was injected into the first 2 groups.All the rats were killed at the 24 h after operation.The levels of TNF-α,IL-1β,NGF,BDNF were measured by enzyme linked immunosorbent assay and the expressions of TNF-α,IL-13,NGF,BDNF mRNA were analyzed by reverse transcription-polymerase chain reaction.Results The contents of TNF-α,IL-1β,NGF,BDNF and their mRNA expressions in hypoxic-ischemic group were significantly higher than those in the sham-operated group.In pretreatment groups,the levels of TNF-α,IL-1β and their mRNA expressions were significantly lower than those in hypoxic-ischemic group.The levels of NGF,BDNF and their mRNA expressions in pretreatment group were significantly higher than those in hypoxic-ischemic group (all P ＜ 0.05).Conclusions Progesterone exerts neuroprotective effect on hypoxic-ischemic encephalopathy-induced brain damage,and the action mechanism is related to down-regulate the expression of damage factor and up-regulate the expression of anti damage factor.

Exploration on the establishment of animal models for gouty nephropathy complicated with chronic renal failure / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the method for establishing animal models of gouty nephropathy complicated with chronic renal failure.</p><p><b>METHODS</b>Six-eight weeks old male Wistar rats were fed with 10% fodder yeast. The adenine at the daily dose of 100, 150, 200, 250, and 300 mg/kg was administrated to them by gastrogavage. The serum levels of blood urea nitrogen (BUN), creatinine (Cr), and uric acid (UA) were dynamically monitored. Meanwhile, the pathological changes of rat kidney were observed.</p><p><b>RESULTS</b>Compared with the normal control group, serum BUN, Cr, and UA obviously increased in rats administered with 100 mg/kg for 7 days (P<0.05). Meanwhile, pathological changes as gouty nephropathy occurred. Along with the prolongation of the modeling time, the aforesaid biochemical indices and pathohistological changes of the kidney were more obvious. The blood Cr level just reached the chronic renal failure level on the 26th day of the administration (about the 4th week), and obviously exceeded the renal failure level on the 41st day (about the 6th week). The blood UA level increased to a higher level on the 7th day of modeling, and maintained at a higher level for a long time. It decreased rapidly from the 41st day to the 48th day. The renal pathological examination showed aggravated infiltration of lymphocytes and stromal fibrous proliferation. On the 48th day of modeling, the proliferation of the fibrous tissue and the interstitial fibrosis were obvious on the bases of the aforesaid changes. The serum BUN, Cr, and blood UA obviously increased in the rats administered with 150, 200, 250, and 300 mg/kg when compared with the normal control group, reaching the level of chronic renal failure (P<0.05). These levels obviously decreased 17 days after restoring to normal fodder feeding, and approached the normal levels till the 35th day.</p><p><b>CONCLUSION</b>Ideal experimental animal models of gouty nephropathy complicated with chronic renal failure could be established in male Wistar rats by feeding with 10% fodder yeast and 100 mg/kg adenine by gastrogavage for 5 weeks.</p>

Prediction model of health workforce and beds in county hospitals of Hunan by multiple linear regression / 中南大学学报(医学版)

OBJECTIVE@#To construct prediction model for health workforce and hospital beds in county hospitals of Hunan by multiple linear regression.@*METHODS@#We surveyed 16 counties in Hunan with stratified random sampling according to uniform questionnaires,and multiple linear regression analysis with 20 quotas selected by literature view was done.@*RESULTS@#Independent variables in the multiple linear regression model on medical personnels in county hospitals included the counties' urban residents' income, crude death rate, medical beds, business occupancy, professional equipment value, the number of devices valued above 10 000 yuan, fixed assets, long-term debt, medical income, medical expenses, outpatient and emergency visits, hospital visits, actual available bed days, and utilization rate of hospital beds. Independent variables in the multiple linear regression model on county hospital beds included the the population of aged 65 and above in the counties, disposable income of urban residents, medical personnel of medical institutions in county area, business occupancy, the total value of professional equipment, fixed assets, long-term debt, medical income, medical expenses, outpatient and emergency visits, hospital visits, actual available bed days, utilization rate of hospital beds, and length of hospitalization.@*CONCLUSION@#The prediction model shows good explanatory and fitting, and may be used for short- and mid-term forecasting.

Effects of activation of liver X receptor and peroxisome proliferator-activated receptor alpha on bile acid synthesis in rats / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the effects of the simultaneous activation of liver X receptor (LXR) and peroxisome proliferator-activated receptor alpha (PPARalpha) on bile acid biosynthesis in rats.</p><p><b>METHODS</b>Totally 36 male SD rats were divided into three groups with 12 rats in each group: control group, high cholesterol (HC) group, and high cholesterol + fenofibrate (HC + FENO) group. Total bile acids (serum bile acids plus fecal bile acids) level was assayed. The levels of mRNA for peroxisomal palmitoyl-CoA oxidase (Acox1), LXR, cholesterol 7alpha-hydroxylase (CYP7A1), D-bifunctional protein (DBP), trihydroxycoprostanoyl-CoA oxidase (Acox2), sterol 12alpha-hydroxylase (CYP8B1), and sterol 27-hydroxylase (CYP27A1) in liver were detected by RT-PCR.</p><p><b>RESULTS</b>Total bile acid level was significantly higher in HC + FENO group than in HC group (P < 0.01), and both were significantly higher than that in control group (P < 0.01). Compared with HC group, the mRNA expression of Acox1 and DBP was significantly higher in HC + FENO group (P < 0.01), but no statistical differences was found between HC group and control group. The mRNA levels of LXR and CYP7A1 in HC + FENO group and HC group were not significantly different but were both significantly higher than that in control group (P < 0.01, P < 0.05). No changes were observed in Acox2, CYPSB1, and CYP27A1 mRNA levels among these three groups.</p><p><b>CONCLUSION</b>Simultaneous activation of LXR and PPARalpha can increase of CYP7A1 and DBP mRNA exDression and thus accelerates the biosynthesis of bile acid.</p>

Relationship between the increase of hepatic D-bifunctional protein activity and bile acid biosynthesis in rats / 中国医学科学院学报

<p><b>OBJECTIVE</b>To determine the physiological role of D-bifunctional protein (DBP) in bile acid biosynthesis through investigating the effect of increasing activity of DBP on bile acid biosynthesis.</p><p><b>METHODS</b>Twenty male Wistar rats were divided into two groups: diethylhexyl phthalate (DEHP) group (n = 10) and control group (n = 10). Serum triglyceride, total cholesterol, hepatic DBP activity, and fecal bile acids were assayed. The mRNA levels of hepatic peroxisome proliferator-activated receptor alpha (PPARalpha), DBP, and cholesterol 7alpha-hydroxylase (CYP7A1) were detected by RT-PCR.</p><p><b>RESULTS</b>Compared with control group, serum triglyceride level was decreased significantly and PPARalphamRNA level was increased significantly in DEHP group (P < 0.01). Together with a sharp induction of DBP mRNA expression and DBP activity in DEHP group (P < 0.01), the levels of CYP7A1 mRNA and fecal bile acids were significantly increased by 1.9 times and 1.6 times respectively compared to control group (P < 0.01). There was a significantly positive correlation between DBP mRNA level or DBP activity and CYP7A1 mRNA level (r = 0.89, P < 0.01; r = 0.95, P < 0.01).</p><p><b>CONCLUSION</b>The up-regulation of DBP mRNA and activity in liver can result in the increase in CYP7A1 mRNA expression and bile acid biosynthesis, suggesting that DBP may be involved in bile acid biosynthesis together with CYP7A1.</p>

Modulation of lianbizi injection (andrographolide) on some immune functions / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of andrographolide on immune functions and the immune mechanism of its clinical application.</p><p><b>METHOD</b>The amounts of IFN-alpha, IFN-gamma, TNF-alpha, IL-8 in the peripheral blood mononuclear cells(PBMCs) culture supernatants deal with by different concentrations of LianBiZhi (LBZ) injection made of andrographolide were detected by biological activity test or ELISA in vitro. The effects of LBZ injection on macrophage phagocytotic function and natural killer cells cytotoxicity were examined by means of macrophage to phagocytize cock erythrocyte and measurement of lactate dehydrogenase(LDH) activity released from the damaged cells, respectively.</p><p><b>RESULT</b>The LBZ injection could promote IFN-alpha, IFN-gamma, TNF-alpha inductions of PBMCs, but had no effect on IL-8. At the same time, the LBZ injection could not only enhance the phagocytosis activity of peritoneal macrophage from guinea pig to phagocytosis cock erythrocyte, but also augment the cytotoxicity mediated by natural killer cells from PBMCs to damage the K562 cell lines.</p><p><b>CONCLUSION</b>Andrographolide is an immunostimulant agent which can modulate both antigen specific and nonspecific immune function by means of its natural killer cells and macrophage and cytokines induction.</p>

Influences of angiotensinⅡtype 1 receptor antagonist on pancreatic steilate cells / 中华消化杂志

To investigate the effects of Losartan,an angiotensinⅡ(AngⅡ)receptor(AT_1) antagonist,on pancreatic stellate cells(PSCs)and its possible mechanisms.Methods (1)PSCs were isolated from pancreatic cancerous samples to test the expressions of AT_1 and collagenⅠafter incubated with AngⅡor/and Losartan.(2)Ninety S-D rats were divided into normal group,control group and treatment group,with 30 rats in each.The rats in control and treatment groups were induced pancreatic fibrosis by injection of 2% trinitrobenenze sulfonic acid(TNBS)into biliopancreatic duct.Rats in treat- ment group were then treated with Losartan by garage daily and rats in control group were only given distilled water.The rats were sacrificed on day 3,7,14,21 and 28,respectively,and pancreas were removed.The histological abnormalities were observed by electron microscope.The mRNAs of trans- forming growth factor?_1(TGF?_1)and procollagenⅠwere detected by reverse transcription-polymerase chain reaction(RT-PCR).The expression of TGF?_1 and?-smooth muscle actin(?-SMA)proteins was assessed by immunohistochemistry and the level of?-SMA protein was quantified by Western blot. Results In vitro,there existed AT_1 expression in PSCs,and Losartan reduced expression of collagenⅠ.Losartan treatment reversed the histological abnormalities observed by electron microscope,com- pared to treatment with distill water.The expression of?-SMA,TGF?_1 and procollagenⅠwere signifi- cantly higher in the control group than those in normal group and were reduced by Losartan to different extent in treatment group.Conclusion AT_1 antagonist can inhibit the activation and the profibrogenic action of PSCs by blocking AT_1 receptor-mediated pathways.