Du Huo Ji Sheng Tang inhibits Notch1 signaling and subsequent NLRP3 activation to alleviate cartilage degradation in KOA mice.

BACKGROUND: Knee osteoarthritis (KOA) has a complex pathological mechanism and is difficult to cure. The traditional medicine Du Huo Ji Sheng Tang (DHJST) has been used for the treatment of KOA for more than one thousand years, but its mechanism for treating KOA has not been revealed. In our previous study, we confirmed that DHJST inhibited the activation of NLRP3 signaling in rats and humans. In the current study, we aimed to determine how DHJST inhibits NLRP3 to alleviate knee cartilage damage. METHODS: Mice were injected with NLRP3 shRNA or Notch1-overexpressing adenovirus into the tail vein to construct systemic NLRP3 low-expressing or Notch1 high-expressing mice. Mice were injected with papain into the knee joint to replicate the KOA model. DHJST was used to treat KOA model mice with different backgrounds. The thickness of the right paw was measured to evaluate toe swelling. The pathohistological changes and the levels of IL-1ß, MMP2, NLRP3, Notch1, collagen 2, collagen 4, HES1, HEY1, and Caspase3 were detected by HE staining, ELISA, immunohistochemical staining, western blotting, or real-time qPCR. RESULTS: DHJST reduced tissue swelling and serum and knee cartilage IL-1ß levels, inhibited cartilage MMP2 expression, increased collagen 2 and collagen 4 levels, decreased Notch1 and NLRP3 positive expression rates in cartilage, and decreased HES1 and HEY1 mRNA levels in KOA model mice. In addition, NLRP3 interference decreased cartilage MMP2 expression and increased collagen 2 and collagen 4 levels without affecting the expression levels of notch1, HES1 and HEY1 mRNA levels in the synovium of KOA mice. In KOA mice with NLRP interference, DHJST further reduced tissue swelling and knee cartilage damage in mice. Finally, Notch1-overexpressing mice not only showed more severe tissue swelling and knee cartilage degradation but also abolished the therapeutic effect of DHJST on KOA mice. Importantly, the inhibitory effects of DHJST on the mRNA expression of NLRP3, Caspase3 and IL-1ß in the knee joint of KOA mice were completely limited after Notch1 overexpression. CONCLUSION: DHJST significantly reduced inflammation and cartilage degradation in KOA mice by inhibiting Ntoch1 signaling and its subsequent NLRP3 activation in the knee joint.

[A new dihydrostilbene from aerial part of Cannabis sativa].

The present study investigated the chemical constituents in the aerial part of Cannabis sativa. The chemical constituents were isolated and purified by silica gel column chromatography and HPLC and identified according to their spectral data and physicochemical properties. Thirteen compounds were isolated from the acetic ether extract of C. sativa and identified as 3',5',4″,2-tetrahydroxy-4'-methoxy-3-methyl-3″-butenyl p-disubstituted benzene ethane(1), 16R-hydroxyoctadeca-9Z,12Z,14E-trienoic acid methyl ester(2),(1'R,2'R)-2'-(2-hydroxypropan-2-yl)-5'-methyl-4-pentyl-1',2',3',4'-tetrahydro-(1,1'-biphenyl)-2,6-diol(3), ß-sitosteryl-3-O-ß-D-glucopyranosyl-6'-O-palmitate(4), 9S,12S,13S-trihydroxy-10-octadecenoate methyl ester(5), benzyloxy-1-O-ß-D-glucopyranoside(6), phenylethyl-O-ß-D-glucopyranoside(7), 3Z-enol glucoside(8), α-cannabispiranol-4'-O-ß-D-glucopyranose(9), 9S,12S,13S-trihydroxyoctadeca-10E,15Z-dienoic acid(10), uracil(11), o-hydroxybenzoic acid(12), and 2'-O-methyladenosine(13). Compound 1 is a new compound, compound 3 is a new natural product, and compounds 2, 4-8, 10, and 13 were isolated from Cannabis plant for the first time.

Increased Consumption of Sulfur Amino Acids by Both Sows and Piglets Enhances the Ability of the Progeny to Adverse Effects Induced by Lipopolysaccharide.

This study determined the effects of increased consumption of sulfur amino acids (SAA), as either DL-Met or Hydroxy-Met (OH-Met), by sows and piglets on their performance and the ability of the progeny to resist a lipopolysaccharide (LPS) challenge. Thirty primiparous sows were fed a diet adequate in SAA (CON) or CON + 25% SAA, either as DL-Met or OH-Met from gestation day 85 to postnatal day 21. At 35 d old, 20 male piglets from each treatment were selected and divided into 2 groups (n = 10/treatment) for a 3 × 2 factorial design [diets (CON, DL-Met or OH-Met) and challenge (saline or LPS)]. OH-Met and/or DL-Met supplementation increased (p ≤ 0.05) piglets' body weight gain during day 0-7 and day 7-14. Sow's milk quality was improved in the supplemented treatments compared to the CON. The LPS challenge decreased (p ≤ 0.05) piglets' performance from 35 to 63 d and increased (p ≤ 0.05) the levels of aspartate aminotransferase, total bilirubin, IL-1ß, IL-6, TNF-a, and malondialdehyde. Plasma albumin, total protein, total antioxidant capacity and glutathione peroxidase decreased post-challenge. The results were better with OH-Met than DL-Met. The increase of Met consumption, particularly as OH-Met increased piglets' growth performance during the lactation phase and the challenging period.

[Microbial Population Dynamics During Sludge Granulation in a Simultaneous Nitrogen and Phosphorus Removal System].

In this study, domestic sewage was utilized to cultivate aerobic granular sludge (AGS) in a simultaneous nitrogen and phosphorus removal (SNPR) system. The bacterial population dynamics during the aerobic sludge granulation were investigated to reveal the granulation mechanisms using Illumina MiSeq PE300 high-throughput sequencing. Quantitative real time polymerase chain reactions (PCR) were used to investigate shifts in the abundance of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), nitrite-oxidizing bacteria (NOB) and polyphosphate accumulating organisms (PAOs). After cultivation for 100 d, the AGS was compact and demonstrated good SNPR performance. During the AGS formation process, extracellular polysaccharides obviously increased, while extracellular proteins kept relatively stable. The abundance of AOA significantly decreased during the formation of AGS process, while the abundance of PAOs increased. The bacterial diversity increased at first and then decreased during the formation of AGS. The bacterial community changed dramatically during aerobic sludge granulation. Persistent operational taxonomic units (OTUs) accounted for 92.70% of the total sequences. Proteobacteria (31.07%-53.67%), Bacteroidetes (6.70%-16.50%) and Chloroflexi (7.84%-13.36%) were the dominant phyla. Candidatus competibacter was obviously enriched in the AGS formation process (increased from 0.11% in the seed sludge to 35.33% in the AGS) and may play an important role in the formation of AGS.

[Chemical constituents from roots of Ixeris chinensis].

Silica gel column chromatography and preparative HPLC were applied to isolate the chemical constituents from the roots of Ixeris chinensis. Fifteen compounds were isolated and their structures were elucidated by the physicochemical properties and spectral analysis as chinensioide G(1), chinensioide B(2), 10α-hydroxy-guaia-12,6-lactone-3-keton(3), chinensioide C(4), 10α-hydroxy-11ßH-guaia-4(5) -ene-12,6-lactone (5), 3ß, 10α-dihydroxy-11ßH-guaia-11 (13)-ene-12,6-lactone (6), 3ß, 10α-dihydroxy-4ßH, 11ßH-guaia-12,6-lactone(7), 3ß, 10α-dihydroxy-guaia-4 (15), 11 (13) -diene-12, 6-lactone (8), caffeic acid (9), p-hydorxyphenylacetic acid(10), methyl p-hydroxyphenylacetate (11), ethyl p-hydroxyphenylacetate (12), sitosterol (13), daucosterol (14), and ixerin D(15). Compound 1 was new, and 6 and 7 were isolated from I. chinensis for the first time.

[Chemical constituents from Euphorbia lunulata].

The chemical constituents from Euphorbia lunulata was investigated in this paper. Fourteen compounds were isolated and purified by column chromatographies on silica gel and preparative HPLC. Their structures were identified by physiochemical properties and NMR data analysis as lupeol (1), euphol (2), cassipourol(3) , 24-methylenecycloartan-3beta-ol (4), 24-hydroperoxycycloart-25-en-3beta-ol (5), 25-hydroperoxycycloart-23-en-3beta-ol (6), betulin (7), uvaol (8), (23E) -25-methoxycycloart-23-en-3beta-ol (9), (23E) -cycloart-23,25-dien-3beta-ol (10), 24-methylenecycloartan-3beta, 28-diol (11), salicinolide (12), 2alpha, 3beta, 5alpha, 9alpha, 15beta-pentaacetoxy-11,12-epoxy-7beta, 8alpha-diisobutyryloxyjatropha-6 (17) -en-14-one (13) and 3beta, 5alpha, 15beta-triacetoxy-7beta-isobutyryloxy-9alpha-nicotinoyloxyjatropha-6 (17), 11(E)-dien-14-one (14). Among them, compounds 1-11 were isolated from E. lunulata for the first time.

Inheritance of hetero-diploid pollen S-haplotype in self-compatible tetraploid Chinese cherry (Prunus pseudocerasus Lindl).

The breakdown of self-incompatibility, which could result from the accumulation of non-functional S-haplotypes or competitive interaction between two different functional S-haplotypes, has been studied extensively at the molecular level in tetraploid Rosaceae species. In this study, two tetraploid Chinese cherry (Prunus pseudocerasus) cultivars and one diploid sweet cherry (Prunus avium) cultivar were used to investigate the ploidy of pollen grains and inheritance of pollen-S alleles. Genetic analysis of the S-genotypes of two intercross-pollinated progenies showed that the pollen grains derived from Chinese cherry cultivars were hetero-diploid, and that the two S-haplotypes were made up of every combination of two of the four possible S-haplotypes. Moreover, the distributions of single S-haplotypes expressed in self- and intercross-pollinated progenies were in disequilibrium. The number of individuals of the two different S-haplotypes was unequal in two self-pollinated and two intercross-pollinated progenies. Notably, the number of individuals containing two different S-haplotypes (S1- and S5-, S5- and S8-, S1- and S4-haplotype) was larger than that of other individuals in the two self-pollinated progenies, indicating that some of these hetero-diploid pollen grains may have the capability to inactivate stylar S-RNase inside the pollen tube and grow better into the ovaries.

Polymorphisms in luteinizing hormone receptor and hypothalamic gonadotropin-releasing hormone genes and their effects on sperm quality traits in Chinese Holstein bulls.

Genes of hypothalamic-pituitary-gonadal axis play a key role in male reproductive performance. This study evaluated the polymorphisms of luteinizing hormone receptor (LHR) and hypothalamic gonadotropin-releasing hormone (GnRH) genes and their effects on sperm quality traits including semen volume per ejaculate (VOL), sperm density (SD), fresh sperm motility (FSM), thawed sperm motility (TSM), acrosome integrity rate (AIR), and abnormal sperm rate (ASR) collected from 205 Chinese Hostein bulls. The study bulls consisted of 205 mature Chinese Holstein, 27 Simmental, 28 Charolais, and 14 German yellow cattle. One single nucleotide polymorphism (SNP) (A883G) in exon 2 of GnRH and two SNPs (A51703G and G51656T) in intron 9 of LHR were identified in 274 bulls. Analysis of variance in 205 Chinese Holstein bulls showed that age had significant effect on both SD and FSM (P < 0.01), and ASR (P < 0.05). With regards to genotype and its interaction with age, only the SNP of G51656T in LHR gene had significant effect on SD (P < 0.05, P < 0.01; respectively). The association result showed that bulls with AG genotype had higher FSM than bulls with AA and GG genotype in LHR at 51,703 locus (P < 0.10), and bulls with GG genotype had higher SD than bulls with TT genotype in LHR at G51656T locus (P < 0.10). Phenotypic correlation among the traits revealed that significant negative correlations were observed between ASR and AIR (r = -0.736, P < 0.01), ASR and AIR (r = -0.500, P < 0.01). There were moderate positive correlations between VOL and SD (r = 0.422, P < 0.01), as well as FSM (r = 0.411, P < 0.01). In conclusion, LHR may be a potential marker for sperm quality of SD and FSM.

[Study on chemical constituents of the Ixeris chinensis].

OBJECTIVE: To study the chemical constituents of the Ixeris chinensis . METHODS: The constituents were isolated by silica gel column chromatography, HPLC and recrystallization and their structures were elucidated on the basis of spectral analysis. RESULTS: Fifteen compounds were isolated and identified as Methyl-4-hydroxyphenylacetate (1), Daucosterol (2), Sitosterol (3), Luteolin-7-O-beta-D-glucoside (4), 15-hydroxy-2-oxoguaia-1 (10), 3,11 (13)-triene-12,6-lactone (5), Chinensiolide B (6), Chinensiolide E (7), Ixerochinoside (8), Chinensiolide C (9), 10alpha-hydroxy-guaia-4(15)-ene-12,6-lactone (10), 10alpha-hydroxy-guaia-4 (15), 11 (13)-diene-12,6-lactone-3beta-O-beta-D-(6'-p-hydroxyphenylacetyl) glucopyranoside (11), Epiloliolide (12), Apigenin-O-beta-D-glucopyranoside (13), Luteolin (14), Lutein (15). CONCLUSION: Compounds 1,10,11,12 and 15 are isolated from this plant for the first time.

Effects of raspberry phytochemical extract on cell proliferation, apoptosis, and serum proteomics in a rat model.

The red raspberry extract possesses potent antioxidant capacity and anticancerous activity in vitro and in vivo. The objective of this study was to determine whether red raspberry extract affected the cell cycle, angiogenesis, and apoptosis in hepatic lesion tissues from a rat model induced by diethylnitrosamine (DEN) as well as changes of serum proteomics. Rats were treated with red raspberry extract (0.75, 1.5, or 3.0 g/kg of body weight) by gavage starting 2 h after DEN administration and continued for 20 wk. Red raspberry extract inhibited cell proliferation, vascular endothelial growth factor VEGF expression, and induced apoptosis in the hepatic lesion tissues. In addition, 2 protein peaks (2597.93 and 4513.88 m/z) were identified to differentially express in the 3.0 g/kg body weight and positive control groups by serum proteomics. These results suggest that a dietary supplement with red raspberry effectively protects against chemically induced hepatic lesions in rats.

[Development and usage of the convenient moxa cone maker].

Moxibustion therapy is a non-invasive and natural therapy that is accepted by modern society with advantages of effectiveness and low-cost. However, the traditional process of producing hand-made moxa cone is time consuming, complicated, inefficient and difficult for mass production. Consequently, it hardly meets the clinical demands. This seriously reduces the popularization and promotion of traditional therapy with moxa cone. To deal with this problem, a new type of convenient moxa cone maker, which can easily and quickly produce multi-model moxa cones in mass production with constant specifications and uniform density. The mass production process is saving in material and low cost with high production efficiency. Application of the convenient moxa cone maker meets the needs of moxa cone in the clinical and scientific research and contributes to the popularization and promotion of traditional therapy with moxa cone.

[Studies on the chemical constituents of Pharbitis purpurea].

OBJECTIVE: To study the chemical constituents of Pharbitis purpurea. METHODS: The constituents were isolated by silica gel column chromatography, HPLC and recrystallization and their structures were elucidated on the basis of spectral analysis. RESULTS: Fourteen compounds were isolated and identified as daucosterol (1), umbelliferone (2), ursolic acid (3), N-p-hydroxy-cis-coumaroyltyramine (4), N-p-hydroxy-trans-coumaroyltyramine (5), N-cis-feruloyltyramine (6), N-trans-feruloyltyramine (7), (3R, 5R, 6S, 7E, 9S)-megastigman-5,6-epoxy-7-ene-3,9-diol (8), (6S,9R)-vomifoliol (9), (+)-syringaresinol (10), isovitexin (11), syringopicroside( 12), uricil (13), (6S,9R)-roseoside (14). CONCLUSION: Compounds 3, 8-2,14 are isolated from the genus for the first time.

[Study on the chemical constituents from fresh roots of Euphorbia fischeriana].

OBJECTIVE: To study the chemical constituents in ethyl acetate fraction from the Euphorbia fischeriana. METHODS: The compounds were isolated by silica gel column chromatography and HPLC, and their structures were elucidated by means of spectral analyses. RESULTS: Fourteen compounds were identified as 2,4-dihydroxy-6-methoxy-3-methylacetophenone (1), 12-deoxyphorbol-13-hexadecanoate (2), ethylgallate (3), esculetin (4), 2,4-dihydroxy-6-methoxy-acetophenone (5), 12-deoxyphorbol-13-acetate (6), 12-deoxyphorbol-13,20-diacetete (7), jolkinolide B (8), 17-hydroxyjolkinolide B (9), 17-hydroxy-jolkinolide A (10), langduin C (11), 3-methylp-hydroxybenzoic acid (12), methyl gallate (13), 3, 3'-diacetyl-pyridine-4,4'-dimethoxy-2, 2', 6, 6'-tetrahydroxydiphenylmethane (14). CONCLUSION: Compounds 3, 5, 12, 13 are isolated from this plant for the first time.

[Autotrophic nitrogen removal and enhanced biological phosphorus removal from municipal wastewater in a three-sludge system].

Using a three-sludge system consisted of anaerobic/oxic (A/O) process, partial nitritation and anaerobic ammonium oxidation (ANAMMOX) reactors, cost-effective removal of nitrogen and phosphate from municipal wastewater was achieved. The experimental results showed that effluent total phosphorus (TP) of the A/O system was less than 0.5 mg/L under hydraulic retention time (HRT) of 3.6 h. Partial nitritation with nitrite accumulation efficiency of 75% -96% was realized in the partial nitritation system under room temperature, DO < 0.2 mg/L and HRT of 4.6 h. Under temperature of 27-30 degrees C and HRT of 1.4 h, effluent total nitrogen (TN) and TN removal rate of ANAMMOX reactor were less than 8 mg/L with the minimum value of 1.6 mg/L and 0.57 kg/(m3 x d), respectively. In the three-sludge system, phosphate accumulating organisms, ammonia-oxidizing bacteria and Anammox bacteria existed under suitably environmental condition to optimize the microbial community structure and improve treatment efficiency of various units. Autotrophic nitrogen removal can reduce 62.5% of the oxygen supply, save 100% of denitrification carbon sources theoretically, lower the sludge production, and greatly decrease carbon dioxide emission. As compared to traditional biological nutrient removal process, the three-sludge system has great advantages and potential in energy saving and carbon dioxide emission reduction to realize sustainable development of water resources.

[Multi-central randomized controlled investigation on the massage for supplementing qi and removing obstruction in the Governor Vessel for treatment of infantile diarrhea due to spleen deficiency].

OBJECTIVE: To make multi-central clinical evaluation of the massage for supplementing qi and removing obstruction in the Governor Vessel for treatment of infantile diarrhea due to spleen deficiency. METHODS: By using multi-central, randomized and controlled method, 275 cases were randomly divided into an observation group (n = 137) and a control group (n = 138). The observation group were treated by the massage for supplementing qi and removing obstruction in the Governor Vessel, and the control group by routine massage therapy in Tuina Science, a teaching material for college and school of TCM. After treatment for 7 days, their therapeutic effects were compared. RESULTS: The cured rate was 83.2% in the observation group and 69.6% in the control group with a signifi cant difference between the two groups (P < 0.05), the former being better than the latter. The mean cured time was (3.22 +/- 1.04) days in the observation group and (4.20 +/- 1.11) days in the control group with a significant difference between the two groups (P < 0.05), the former being shorter than the latter. CONCLUSION: The massage for supplementing qi and removing obstruction in the Governor Vessel has a definite therapeutic effect on infantile diarrhea due to spleen deficiency, with rapid effect.

[On moxibustion for preventing diseases].

Along with the improvement of living standard, people's consciousness about health care is gradually raising. Moxibustion is a convenient and effective therapy for preventing diseases and prolonging life. In this paper, functions of moxibustion therapy are probed from the preventing diseases, reasonable selection of acupoints and clinical studies, etc. It is held that moxibustion can promote metabolism and increase immune function, hence preventing diseases and prolonging life.

[Studies on chemical constituents of Astragalus dahuricus].

OBJECTIVE: To study the chemical constituents in Astragalus dahuricus. METHOD: The compounds were isolated and purified by column chromatography on silica gel and HPLC, and their structures were elucidated by their spectroscopic evidences. RESULT: Six compounds were identified as: 7, 2'-dihydroxy-3', 4'-dimethoxyisoflavan (1), 2'-hydroxy-3', 4'-dimethoxyisoflavan-7-O-beta-D-glucopyranoside (2), 8, 2'-dihydroxy-7, 4'-dimethoxyisoflavan (3), 7-hydroxy-4'-methoxyisoflavone (4), 7, 3'-dihydroxy-4'-metho-xyisoflavone (5), 9, 10-dimethoxypterocarpan-3-O-beta-D-glucopyranoside (6). CONCLUSION: Compounds 1-6 were obtained from this plant for the first time.

[Chemical constituents from Cirsium pendulum Fisch. ex DC].

To study the chemical constituents of Cirsium pendulum Fisch. ex DC. Thirteen compounds were obtained from ethanol extract of Cirsium pendulum Fisch. ex DC. These compounds were characterized by spectroscopic analysis and comparison with published data to be beta-sitosterol(1), alpha-spinastenorl(2), taraxasterol(3), triacontanol(4), chlorophylla(5), lutein(6), linarin(7), 3-sitosterol-3-O-beta-D-glucopyranoside (8), 3', 4'-dihydroxy phenylethanol glucoside(9),3-(4'-methoxy-3',5'-dihydroxyphenyl)-allyl alcohol glucoside (10), uracil (11), methyl chlorogenate (12), chlorogenic (13). All of them were isolated from this plant for the first time.