Objective: To observe the effect of modified Erchentang on the expressions of NLRP3 inflammasome genes in peripheral blood mononuclear cells (PBMCs) and the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)and chemokine8 (CXCL8) in lung tissue of rats with chronic obstructive pulmonary disease (COPD), in order to explore the molecular mechanism of modified Erchentang against inflammation of COPD. Method: Forty SD rats were randomly divided into normal control group, model group, MCC950 (NLRP3 inhibitor) group and modified Erchentang group. The COPD model of rats was prepared by using cigarette smoke and dripping with lipopolysaccharide (LPS). During the modeling period (from the 1^st to the 30^th day), the MCC950 group received a single intraperitoneal injection with 60 mg · kg^-1 on the first day of the experiment,and the modified Erchentang group was given intragastric administration with 10 g · kg^-1, once every 2 days. From the 31^st to the 45^th day, the MCC950 group was intraperitoneally injected with 3 mg · kg^-1, once every 2 days, the modified Erchentang group was given intragastric administration with 10 g · kg^-1, twice a day, and the normal group and the model group received normal saline (NS) with 10 g · kg^-1, twice a day. The levels of interleukin-1β(IL-1β), interleukin-18(IL-18) and chemokine8 (CXCL8) in rats lung tissue homogenate were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of NLRP3, apoptosis-associated speck-like protein (ASC) and cysteinyl aspartate specific proteinase-1 (Caspase-1) mRNA in PBMCs were measured by Real-time fluorescence quantitative PCR (Real-time PCR). Western blot was used to detect the levels of NLRP3, ASC and Caspase-1 proteins in PBMCs. Immunohistochemical(IHC)method was used to detect the expressions of NLRP3, ASC and Caspase-1 proteins in lung tissues. Result: The expressions of NLRP3, ASC and Caspase-1 mRNA and protein were increased significantly (P<0.01) in model group compared with normal group. Compared with model group, the expressions of NLRP3, ASC and Caspase-1 mRNA and protein were decreased significantly in PBMCs (P<0.05,P<0.01) of modified Erchentang group. The levels of IL-18, IL-1β and CXCL8 in lung tissue homogenate in model group were significantly higher than those in the control group. However, compared with model group, the levels of IL-18, IL-1β and CXCL8 were decreased significantly (P<0.05,P<0.01) in modified Erchentang group. Conclusion: NLRP3 inflammasome is involved in the inflammatory response in COPD rats. Modified Erchentang may inhibit the inflammatory response of COPD effectively. The mechanism may be correlated with the reduction of NLRP3, ASC and Caspase-1 gene expressions, and the inhibition of the release of IL-18, IL-1β and CXCL8.

The research is aimed to study of the influence of environmental factors on the yield and quality traits, and find out the regularity of the growth and development of perilla. The main environmental factor data in six ecological area in Guizhou province were collected, and the correlation analysis with yield and quality traits of 15 perilla strains was conducted. The results showed that the cultivation environment has significant effects on the yield and quality traits of perilla. The effect of environment on main yield composed traits, contained grain number in top spike, effective panicle number per plant, plant height, top spike length, growth period, and thousand seed weight was degressive. In the different environmental factors, the latitude showed positive correlation with yield, growth period and effective panicle number per plant, and negative correlation with top spike length and grain number in top spike. Elevation showed negative correlation with the growth period of perilla. The perilla yield increased at first and then decreased with altitude rising, with the maximum in the 800 m altitude. The 600-900 m altitude is suitable area for perilla. Except for positive correlation with the plant height, and negative correlation with top spike length, the longitude showed in apparent impact on other traits. Sunshine duration, temperature and rainfall accumulation showed different effect on the different perilla strains. For yield composed traits, the sunshine duration was negatively correlation with the plant length. The accumulated temperature and mean temperature showed negative correlation with the main spike length, the rainfall showed negative correlation with the precipitation and growth period, plant height, ear number. The environmental impact on the oil compounds decreased with oleic acid, stearic acid, linoleic acid, -linolenic acid, palmitic acid and oil content. Correlation analysis showed that the significantly negative correlation between the oil content and palmitic acid and linoleic acid content, and the positive correlation between linolenic acid content, -linolenic acid content showed significant negative correlation with other fatty acids composition, and palmitic acid, stearic acid, oleic acid, linoleic acid showed significant positive correlation with each other. The influence of different environmental factors on the quality of perilla were as follows: the oil content was positively associated with elevation and sunshine duration. -Linolenic acid content showed negative correlation with longitude, latitude, accumulated temperature and mean temperature, but positive correlation with altitude, sunlight and rainfall capacity. The correlation between palmitic acid, stearic acid, oleic acid, linoleic acid and environmental factors showed contrast character of -linolenic acid. This study detailed discussed the influence of environmental factors on the quality of perilla, which provided the foundation of ecological planting technology and geoherbalism research of perilla.
Environment ; Fatty Acids ; analysis ; Perilla frutescens ; chemistry ; growth & development ; Phytochemicals ; analysis ; Plant Oils ; analysis

To study the pharmacokinetic effect of Mori Folium flavones and alkaloids in normal and diabetic rats. An UPLC-TQ-MS method was developed for the simultaneous determination of rutin, isoquercitrin, astragalin, kaempferol, quercetin, chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, DNJ and fagomine in plasma of rats. The diabetic rat model was induced through intravenous injection with alloxan and high-fat diet. Samples of plasma of rats were obtained at different time points, after the rats were administrated with Mori Folium flavones and alkaloids. After the deproteinization with acetonitrile, the concentrations of Mori Foliam constituents in rats at different time points were detected by UPLC-TQ-MS method, and pharmacokinetic parameters were calculated by DAS 2.0 software. The results showed that quercetin and kaempferol reached peak at 0.333 h, indicating that Mori Folium flavonoid constituents were absorbed and distributed quickly. At about 4 h after administration, both of them reached the peak concentrations for the second time, suggesting that they stayed in intestine for a long time. DNJ and fagomine in gastrointestinal tract can be quickly absorbed into blood, and the concentration in plasma reached peak after 0.667 h, suggesting that both of them could be rapidly distributed in the systemic circulation of rats. Cryptochlorogenic acid, neochlorogenic acid, quercetin, kaempferol and rutin were found to have a higher Cmax and AUC0-t in normal rats than those in diabetic rats. The t1/2values of cryptochlorogenic acid and neochlorogenic acid were shorter in diabetic rats, while quercetin, kaempferol and rutin had a longer t1/2value in diabetic rats. Chlorogenic acid, astragalin, isoquercitrin, fagomine had a higher Cmax in diabetic rats, and the t1/2values of astragalin and fagomine were longer, which suggested differences in absorption of active ingredients under normal and diabetic conditions.

Microglia play a pivotal role in clearance of Aβ by degrading them in lysosomes, countering amyloid plaque pathogenesis in Alzheimer's disease (AD). Recent evidence suggests that lysosomal dysfunction leads to insufficient elimination of toxic protein aggregates. We tested whether enhancing lysosomal function with transcription factor EB (TFEB), an essential regulator modulating lysosomal pathways, would promote Aβ clearance in microglia. Here we show that microglial expression of TFEB facilitates fibrillar Aβ (fAβ) degradation and reduces deposited amyloid plaques, which are further enhanced by deacetylation of TFEB. Using mass spectrometry analysis, we firstly confirmed acetylation as a previously unreported modification of TFEB and found that SIRT1 directly interacted with and deacetylated TFEB at lysine residue 116. Subsequently, SIRT1 overexpression enhanced lysosomal function and fAβ degradation by upregulating transcriptional levels of TFEB downstream targets, which could be inhibited when TFEB was knocked down. Furthermore, overexpression of deacetylated TFEB at K116R mutant in microglia accelerated intracellular fAβ degradation by stimulating lysosomal biogenesis and greatly reduced the deposited amyloid plaques in the brain slices of APP/PS1 transgenic mice. Our findings reveal that deacetylation of TFEB could regulate lysosomal biogenesis and fAβ degradation, making microglial activation of TFEB a possible strategy for attenuating amyloid plaque deposition in AD.
Alzheimer Disease ; metabolism ; pathology ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; chemistry ; genetics ; metabolism ; Brain ; metabolism ; Cells, Cultured ; Chloride Channels ; genetics ; metabolism ; Disease Models, Animal ; HEK293 Cells ; Humans ; Lysosomes ; genetics ; metabolism ; Mice ; Mice, Transgenic ; Microglia ; cytology ; metabolism ; Mutagenesis, Site-Directed ; Peptides ; analysis ; chemistry ; Protein Binding ; RNA Interference ; Sirtuin 1 ; antagonists & inhibitors ; genetics ; metabolism

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.
Alkaloids ; chemistry ; Amino Acids ; metabolism ; Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; drug therapy ; Flavones ; chemistry ; Flavonoids ; chemistry ; Metabolic Networks and Pathways ; Metabolomics ; Mice ; Morus ; chemistry ; Plant Leaves ; chemistry ; Sphingolipids ; metabolism ; Tandem Mass Spectrometry

OBJECTIVETo analyze the correlation of Chinese medicine syndrome evolvement and cardiovascular: events in patients with stable coronary heart disease (CHD).

METHODSThis prospective cohort study investigated and: collected the clinical information of patients with stable CHD and observed the syndrome type at the baseline and 6-month at follow-up, as well as the cardiovascular events during the 6-month and 12-month follow-up. The patients were divided into the event group and the non-event group. The interaction and the impact of syndrome evolvement on cardiovascular events were examined through multifactor dimensionality reduction (MDR) analysis and the results were verified by Chi-square test.

RESULTSTotally 1,333 of 1,503 stable CHD patients enrolled met the inclusion criteria: of MDR analysis. Among them, 959 (71.9%) cases were males and 374 (28.1%) cases were females. Thirty seven cases had cardiovascular events during 6 to 12 months after the study began. The results of the MDR analysis and verification using Chi-square test showed that the development of cardiovascular events was positively correlated with interaction between blood stasis and toxic syndrome at the baseline, blood stasis at the baseline and qi deficiency at the 6-month follow-up, toxic syndrome at the baseline and qi deficiency at the 6-month follow-up, toxic syndrome at the base line and blood stasis at the 6-month follow-up, qi deficiency and blood stasis at the 6-month follow-up (P<0.05 for all).

CONCLUSIONSBlood stasis, toxic syndrome and qi deficiency are important factors of stable CHD. There: are positive correlation between cardiovascular events and syndrome evolution from blood stasis to qi deficiency, from toxic syndrome to qi deficiency and from toxic syndrome to blood stasis, indicating the pathogenesis of toxin consuming qi, toxin leading to blood-stasis in stable CHD patients prone to recurrent cardiovascular events.

Aged ; Cardiovascular Diseases ; etiology ; Coronary Angiography ; Coronary Disease ; complications ; physiopathology ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Multifactor Dimensionality Reduction ; Syndrome

OBJECTIVETo investigate retrospectively the clinical effects and recurrence rate of 143 cases who underwent one level discectomy and followed up more than 10 years. To evaluate the outcome of patients in groups of different operating age and extents of disc herniation, and analyse whether difference exists in each group.

METHODSThere were 143 patients (operation time from January 1996 to December 2000) including 80 males and 63 females, aged from 18 to 66 years old with an average of 37.85 years. The followed-up time was 10 to 15 years with an average of 12.7 years. Patients were divided into 3 groups depends on operating age: < 30 years old, 30 to 50 years old and > 50 years old; 87 patients who's pre-operative CT scan could be collected among 143 cases were divided into 3 groups depends on extents of disc herniation: I degree, II degrees, and III degrees. The final followed-up was obtained in 2011, to evaluate each group and the holistic clincal outcome with JOA scores and ODI scores, and observe whether there were difference between every groups; to judge the effects by patient himself with modified Macnab Criteria.

RESULTS(1) JOA scores pre-operation and final followed-up was 5.11 +/- 2.02 and 12.51 +/- 2.35 respectively; ODI scores pre-operation and final followed-up was 33.98 +/- 7.42 and 13.39 +/- 6.79 respectively. There were significant differences between pre-operative and final followed-up in JOA and ODI (P < 0.01). The excellent-good rate was 83.2% (119/143 ) according to modified Macnab Classification with recurrence rate of 6.3% at final follow-up. (2) Obvious difference was found in JOA scores in group who's age at operation less than 30 years old compared with other 2 groups at followed-up time, and no significant difference was found in JOA scores between other two groups ; no significant difference was found in ODI scores among the three groups. (3) Significant difference was found in JOA and ODI scores in group with III degrees lumbar disc herniation group compared with other 2 groups, and no statistical difference was found in clincal scores between other 2 groups.

CONCLUSION(1) Long-term followed-up of 143 cases prove mono-level lumbar discectomy is an option for disc herniation with good curative effect and lower recurrent rate, the technique should be the prior selection in dealing with patients with lumbar disc herniation. (2) 51% patients (19/37) in group under 30 years old endure persistent low back pain. (3) The long-term clinical effects in patients with severe disc protrution who underwent lumbar discectomy is worse than those patients with mild lumbar disc herniation.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc ; surgery ; Intervertebral Disc Displacement ; surgery ; Laminectomy ; methods ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies

BACKGROUNDErythromycin-resistant Streptococcus pneumoniae isolates that causing invasive pneumococcal diseases (IPD) in Chinese children remain uncharacterized. This study aims to identify the resistance genes associated with erythromycin resistance and to determine the genetic relationships of IPD isolates in Chinese children.

METHODSA total of 171 S. pneumoniae strains were isolated from 11 medical centers in China from 2006 to 2008. All the isolates were characterized via serotyping and antibiotic susceptibility determination. The erythromycin-resistant isolates were further characterized via ermB and mefA gene detection, multi-locus sequence typing analysis, and pulsed-field gel electrophoresis.

RESULTSA total of 164 (95.9%) isolates showed resistance to erythromycin, of which 162 strains with high high-level resistance (MIC ≥ 256 µg/ml). A total of 104 (63.4%) isolates carry the ermB gene alone, whereas 59 (36.0%) harbor both ermB and mefA genes. Of the 59 strains, 54 were of serotypes 19A and 19F and were identified as highly clonal and related to the Taiwan(19F)-14 clone.

CONCLUSIONSThe erythromycin resistance rate in IPD isolates is significantly high and is predominantly mediated by the ermB gene. Isolates that carry both ermB and mefA genes are predominantly of serotypes 19A and 19F.

Adolescent ; Anti-Bacterial Agents ; pharmacology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Erythromycin ; pharmacology ; Humans ; Infant ; Multilocus Sequence Typing ; Pneumococcal Infections ; microbiology ; Serotyping ; Streptococcus pneumoniae ; classification ; drug effects ; genetics ; isolation & purification

OBJECTIVETo investigate the hematological characteristics of co-inheritance of α-thalassemia (α-thal) and β-thalassemia (β-thal) and to survey the incidence of co-inheritance of α-thal and β-thal in Guangxi.

METHODSDNA samples from 370 primary and middle school students who were β-thal carriers in Guangxi were further processed for the α-goblin gene mutation screening, and were grouped based on the genotype of β- and α-goblin gene. The hematological indexes to the different groups were compared by One-way ANOVA.

RESULTSOf the total 370 β-thal carriers, 79 were found to carry α-thal, which gave a frequency of 21.35% for β-thal carriers and 1.36% for coincidence of these two common disorders in the local population. As expected, the 79 patients presented very variable α-globin alterations in combination with β-globin mutations, showing 31 genotype combined with the coincidence of both Hb disorders. Except the genotypes of 3 β-thal heterozygotes combined with ααα(anti3.7) triplication and 2 β-thal carriers with IVS-II-654(C→T)/N combined-α(3.7)/αα presented the phenotype of thalassemia intermedia, and other 74 carriers with co-inheritance of α-thal and β-thal all presented the phenotype of β-thal trait. There were significant differences between β-thal heterozygotes and the carriers with a co-inheritance of both β+α(0) thal in MCH, MCV and Hb. In addition, there existed significant difference between the carriers with a co-inheritance of both β+α(+) thal and a co-inheritance of both β+α(0) thal in MCV, MCH and Hb.

CONCLUSIONCompared to that of β-thal heterozygotes, the carriers with a co-inheritance of α-thal and β-thal had slighter phenotype with hematological characteristics. It's difficult to distinguish the double heterozygotes with the co-inheritance of α-thal and β-thal from β-thal heterozygotes by hematological indexes, the molecular diagnosis should be performed.

Child ; China ; epidemiology ; Female ; Gene Frequency ; Genotype ; Humans ; Incidence ; Male ; alpha-Thalassemia ; blood ; epidemiology ; genetics ; beta-Thalassemia ; blood ; epidemiology ; genetics

Objective To analyze the drug resistant characteristics of 84 clinical isolated Helicobacter pylori (Hp) strains, and to observe the inhibitory effects of anti-Hp Lactobacillus acidophilus (La)4 and La6 on different antibiotic-resistant Hp strains. Methods Hp strains were isolated and cultured from gastric mucosa of 84 different gastropathy patients (20 patients with chronic gastritis, 24 with gastric ulcer, 19 with duodenal ulcer and 16 with gastric cancer). The minimum inhibitory concentration (MIC) of metronidazole, clarithromycin and amoxicillin were tested by E-test in order to determine the resistance of these three antibiotics in clinical isolated Hp strains. With standard La as control, the supernatant of anti-Hp La4 and La6 was added into Hp strains culture wells. Hp strains were cultured in solid media for 72 hours, and then inhibition ring were recorded. Anti-Hp Lactobacillus acidophilus liquid was also added to culture medium of different Hp strains, which were in liquid culture, culture medium were taken at different time points (4,8,12,24,48 hrs) to calculate bacteria colony number and test urease activity. Results In 84 clinical isolated Hp strains, the resistant rates of metronidazole, clarithromycin and amoxicillin resistance rates were 67.9%, 17.9% and 1.2% respectively. Of those 11 strains were mixed drug resistance, which included 10 strains of metronidazole and clarithromycin mixed drug resistance, and one of metronidazole and amoxicillin mixed drug resistance. In solid culture conditions, supernatant of anti-Hp Lactobacillus acidophilus La4 and La6 had obvious inhibitory effect on antibiotic-resistant and non-resistant Hp strains. In liquid culture conditions, anti-Hp Lactobacillu acidophilus La4 and La6 bacterium liquid could inhibit the proliferation of antibiotic-resistant and non-resistant Hp strains, the antagonistic role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). The urease activity of antibiotic-resistant Hp strains was inhibited since mixed cultured with anti-Hp Lactobacillu acidophilus La4 and La6 for 4 hours, the urease activity gradually decreased as culture time extended, and the inhibitory role was significantly stronger than the standard Lactobacillus acidophilus strains (P<0.05). Conclusions In 84 Hp strains, most were metronidazole resistant strains, followed by clarithromycin resistant strains, metronidazole and clarithromycin mixed resistance strains. In vitro, anti-Hp Lactobacillu acidophilus La4 and La6 had obvious inhibitory effects on antibiotic-resistant and non-resistant Hp strains.