Anti-inflammatory properties and characterization of water extracts obtained from Callicarpa kwangtungensis Chun using in vitro and in vivo rat models.

Callicarpa kwangtungensis Chun (CK) is a common remedy exhibits anti-inflammatory properties and has been used in Chinese herbal formulations, such as KangGongYan tablets. It is the main component of KangGongYan tablets, which has been used to treat chronic cervicitis caused by damp heat, red and white bands, cervical erosion, and bleeding. However, the anti-inflammatory effects of CK water extract remains unknown. This study assessed the anti-inflammatory effects of CK in vivo and in vitro, characterized its main components in the serum of rats and verified the anti-inflammatory effects of serum containing CK. Nitric oxide (NO), tumour necrosis factor α (TNF-α) and interleukin-6 (IL-6) release by RAW264.7 cells was examined by ELISA and Griess reagents. Inflammation-related protein expression in LPS-stimulated RAW264.7 cells was measured by western blotting. Furthermore, rat model of foot swelling induced by λ-carrageenan and a collagen-induced arthritis (CIA) rat model were used to explore the anti-inflammatory effects of CK. The components of CK were characterized by LC-MS, and the effects of CK-containing serum on proinflammatory factors levels and the expression of inflammation-related proteins were examined by ELISA, Griess reagents and Western blotting. CK suppressed IL-6, TNF-α, and NO production, and iNOS protein expression in LPS-stimulated RAW264.7 cells. Mechanistic studies showed that CK inhibited the phosphorylation of ERK, P38 and JNK in the MAPK signaling pathway, promoted the expression of IκBα in the NF-κB signaling pathway, and subsequently inhibited the expression of iNOS, thereby exerting anti-inflammatory effects. Moreover, CK reduced the swelling rates with λ-carrageenan induced foot swelling, and reduced the arthritis score and incidence in the collagen-induced arthritis (CIA) rat model. A total of 68 compounds in CK water extract and 31 components in rat serum after intragastric administration of CK were characterized. Serum pharmacological analysis showed that CK-containing serum suppressed iNOS protein expression and NO, TNF-α, and IL-6 release. CK may be an anti-inflammatory agent with therapeutic potential for acute and chronic inflammatory diseases, especially inflammatory diseases associated with MAPK activation.

Cayratia albifolia C.L.Li exerts anti-rheumatoid arthritis effect by inhibiting macrophage activation and neutrophil extracellular traps (NETs).

BACKGROUND: Cayratia albifolia C.L.Li (CAC), commonly known as "Jiao-Mei-Gu" in China, has been extensively utilized by the Dong minority for several millennia to effectively alleviate symptoms associated with autoimmune diseases. CAC extract is believed to possess significant anti-inflammatory properties within the context of Dong medicine. However, an in-depth understanding of the specific pharmaceutical effects and underlying mechanisms through which CAC extract acts against rheumatoid arthritis (RA) has yet to be established. METHODS: Twenty-four Sprague-Dawley rats were divided into four groups, with six rats in each group. To induce the collagen-induced arthritis (CIA) model, the rats underwent a process of double immunization with collagen and adjuvant. CAC extract (100 mg/kg) was orally administered to rats. The anti-RA effects were evaluated in CIA rats by arthritis score, hind paw volume and histopathology analysis. Pull-down assay was conducted to identify the potential targets of CAC extract from RAW264.7 macrophage lysates. Moreover, mechanism studies of CAC extract were performed by immunofluorescence assays, real-time PCR and Western blot. RESULTS: CAC extract was found to obviously down-regulate hind paw volume of CIA rats, with diminished inflammation response and damage. 177 targets were identified from CAC extract by MS-based pull-down assay. Bioinformatics analysis found that these targets were mainly enriched in macrophage activation and neutrophils extracellular traps (NETs). Additionally, we reported that CAC extract owned significant anti-inflammatory activity by regulating PI3K-Akt-mTOR signal pathway, and inhibited NETosis in response to PMA. CONCLUSIONS: We clarified that CAC extract significantly attenuated RA by inactivating macrophage and reducing NETosis via a multi-targets regulation.

New Terpenoids from Potentilla freyniana Bornm. and Their Cytotoxic Activities.

Two new A-ring contracted triterpenoids, madengaisu A and madengaisu B, and one undescribed ent-kaurane diterpenoid, madengaisu C, along with 20 known compounds were isolated from the roots of Potentilla freyniana Bornm. The structures were elucidated using extensive spectroscopic techniques, including 1D and 2D-NMR, HR-ESI-MS, ECD spectra, IR, and UV analysis. Moreover, all isolated constituents were evaluated for their anti-proliferative activity against RA-FLS cells and cytotoxic activities against the human cancer cell lines Hep-G2, HCT-116, BGC-823, and MCF-7. Ursolic acid and pomolic acid displayed moderate inhibitory activity in RA-FLS cells with IC50 values of 24.63 ± 1.96 and 25.12 ± 1.97 µM, respectively. Hyptadienic acid and 2α,3ß-dihydroxyolean-12-en-28-oic acid 28-O-ß-d-glucopyranoside exhibited good cytotoxicity against Hep-G2 cells with IC50 values of 25.16 ± 2.55 and 17.66 ± 1.82 µM, respectively. In addition, 2α,3ß-dihydroxyolean-13(18)-en-28-oic acid and alphitolic acid were observed to inhibit HCT-116 cells (13.25 ± 1.65 and 21.62 ± 0.33 µM, respectively), while madengaisu B and 2α,3ß-dihydroxyolean-13(18)-en-28-oic acid showed cytotoxic activities against BGC-823 cells with IC50 values of 24.76 ± 0.94 and 26.83 ± 2.52 µM, respectively, which demonstrated that triterpenes from P. freyniana may serve as therapeutic agents for RA and cancer treatment.

[Research progress on chemical constituents and pharmacological activities of Potentilla].

There are 200-500 species of Potentilla(Rosaceae) worldwide, among which 90 species are widely distributed in China and have a long history of ethnic medicinal use. According to our statistics, a total of 367 compounds have been isolated and identified from plants of this genus, including terpenoids, flavonoids, phenolic acids, tannins, and phenylpropanoids. The medicinal materials made from these plants mainly have antioxidative, blood sugar-lowering, anti-inflammatory, anti-tumor, cardiovascular system-protecting, neuroprotective, and hepatoprotective activities. This study systematically reviews the research progress on chemical constituents and pharmacological activities of Potentilla plants to provide a basis for further research and clinical application.

Interleukin-18 promotes fibroblast senescence in pulmonary fibrosis through down-regulating Klotho expression.

Idiopathic pulmonary fibrosis (IPF) is a lethal disease of unknown aetiology that largely presents in the elderly. The mechanisms related to aging such as fibroblast senescence have been strongly implicated in pathology of IPF. We have previously demonstrated the protective effects of IL-18 binding protein (IL-18BP) against bleomycin (BLM)-induced pulmonary fibrosis (PF) via inhibition of epithelial-to-mesenchymal transition. However, the role of IL-18 in fibroblast senescence in PF is still unknown. The aim of this study was to investigate the effects of IL-18 on fibroblast senescence in the development of PF. We found that SA-ß-gal positive cells, the proportion of cells in G1 phase, and expressions of p21 and p53 were increased in primary lung fibroblasts isolated from BLM-challenged mice, while the fibroblasts from IL-18BP-treated mice showed decreased senescence propensity. We further demonstrated that IL-18 was sufficient to trigger senescence of primary lung fibroblasts. The senescence-associated secretory phenotype (SASP) of fibroblasts treated with IL-18 robustly stimulated a fibrotic phenotype in pulmonary fibroblasts. Moreover, the expression of Klotho, an anti-senescence protein, was down-regulated after IL-18 treatment in primary lung fibroblasts. Overexpression of Klotho reversed the senescence and SASP induced by IL-18 in lung fibroblasts. In summary, we reported for the first time that IL-18 promoted the lung fibroblast senescence and SASP in PF through blocking Klotho pathway. Neutralize IL-18 by IL-18BP exhibited antifibrotic effects partly by suppressing lung fibroblast senescence in PF. It contributes to the growing evidence that IL-18 could be a therapeutic target for PF.

Sema 3A as a biomarker of the activated mTOR pathway during hexavalent chromium-induced acute kidney injury.

Semaphorin 3A (sema 3A) is one of a class of secretory proteins belonging to a family of axon-directed factors found in podocytes, distal tubules, and collecting tubes of the kidney. It is considered to be a potential target molecule involved in the mammalian target of the rapamycin (mTOR) pathway in renal injury or renal diseases, but it has an unknown role in the course of hexavalent chromium-Cr(VI) induced nephrotoxicity. In the present study, an acute kidney injury (AKI) model in rats or cultured tubular epithelial HK-2 cells was employed for Cr(VI) exposure alone or in combination with rapamycin (Rap) or N-acetyl-l-cysteine (NAC) or recombinant sema 3A. The methods of histopathology, biochemics, and western blotting were applied to evaluate tubular injury and the role of sema 3A. The results showed that a significant increase of urinary sema 3A indicates an early occurrence of AKI exposed to Cr(VI), accompanied with a significant increase of tubular injury score and phosphorylated mTOR proteins. Further, Cr(VI) treatment, in combination with pretreatment of the mTOR pathway inhibitor, Rap, showed a considerably stronger protective effect of Rap in protecting against Cr(VI)-induced nephrotoxicity than that seen with the free radical scavenger NAC, highlighting the dominant renal protective role of the mTOR pathway in inhibiting toxicity by downregulating the expressed levels of sema 3A in renal tissue. This study has demonstrated that an increased expression of sema 3A occurs in Cr(VI)-induced AKI resulting from activation of the mTOR pathway, and that inhibition of this pathway has been shown to decrease the severity of the toxicity. In conclusion, this study has shown that increased urinary sema 3A is indicative of an activated mTOR pathway and is a valuable biomarker of the early AKI induced by Cr(VI) exposure.

A de novo 1.38 Mb duplication of 1q31.1 in a boy with hemifacial microsomia, anophthalmia, anotia, macrostomia, and cleft lip and palate.

We reported a 2-year-old boy with developmental delay, mild mental retardation, and severe craniofacial malformation, including facial asymmetry with hypoplasia of the left zygoma, maxilla, and mandible, and left anophthalmia and anotia. A genome-wide screen revealed a 1.38 Mb duplication on chromosome 1q31.1, which was absent in his parents and 27 healthy controls. The duplication region contains two Refseq genes, PLA2G4A and C1orf99, which have not been reported to be implicated in craniofacial malformation. Functional studies of these genes and additional clinical analysis are necessary to elucidate the pathogenesis of craniofacial malformation.

[The epidemiological characteristic of 97,823 cases of pre-hospital medical care in Guangzhou city].

OBJECTIVE: To investigate the epidemiological information of patients in pre-hospital medical care in Guangzhou city, and to explore the characteristics of the patients. METHODS: The data in the year of 2008 were retrieved from the computer database of Guangzhou Emergency Medical Rescue Command Center. RESULTS: (1)In a total of 969 410 calls received, the time of distribution was found to be mainly between 16:00 and 18:00 [11.78% (114 224)], and least frequently between 04:00 and 06:00 [2.40% (23 237)]. (2)Among 109 682 dispatches of ambulances, Baiyun district received the most [26.77% (29 364)], and followed by Haizhu district [18.30% (20 069)], Tianhe district [18.20% (19 962)], respectively. (3)Among 97 823 cases of pre-hospital medical care, death rate of the male patients was higher than the female [amount: 57.65% (56 394) vs. 38.48% (37 641), mortality: 59.17% (3 269) vs. 33.95% (1 876)]. (4)In 9 7823 cases of pre-hospital medical care, trauma constituted the highest rate [34.57% (33 820)], especially traffic accidents [11.56% (11 307)], and the age of most of the patients ranged between 21 and 50. Disease of the nervous system ranged the second, followed by diseases of circulatory system, respiratory system and digestive system, and most of them were over 51 years old, and most frequently above 70. (5)In 97 823 cases of pre-hospital medical care, there were 5 525 deaths (5.65%), in whom the circulatory system diseases ranged first (especially sudden death) [33.07% (1 827)], followed by unclassified diseases [29.79% (1 646)], trauma [15.67% (866)], respiratory diseases [7.48% (413)], and neurological emergency illnesses [5.95% (329)]. The age of deceased was far older than 51, particularly 70. The age of most of the deceased was above 61, and age of traumatic death was 21-40. CONCLUSION: (1) It is very important to reduce the death rate of the middle-old aged patients by strengthening prevention and timely treatment of cardiovascular and cerebrovascular diseases, and improve the medical strategies in emergency care, in order to lower the death rate during emergency.(2)It is very important to emphasize safely in production lines and to strengthen traffic regulations in order to reduce the incidence of trauma, thus it is especially traffic accident, expect that the death rate of trauma could be lowered.

[A randomized controlled trial study of pulmonary rehabilitation with respiratory physiology as the guide on prognosis in patients with chronic obstructive pulmonary disease].

OBJECTIVE: To observe the effect of pulmonary rehabilitation with respiratory physiology as guide in patients with chronic obstructive pulmonary disease (COPD). METHODS: Sixty patients of severe and very severe COPD as categorized by global proposed diagnostic criteria for COPD (GOLD, 2006) were enrolled for study. They were randomly divided into three groups, and with 20 patients in each group. The patients in group A were given pulmonary rehabilitation guided by respiratory physiology thrice a day, 15 minutes each time for 8 weeks. The patients in group B were given pulmonary rehabilitation with pursed lip respiration thrice a day, 15 minutes per time for 8 weeks. The patients in group C were given no pulmonary rehabilitation. Six minute-walk-distance (6MWD), medical research council (MRC) dyspnea scale, activities of daily living (ADL), maximal expiratory pressure (MEP), maximal inspiratory pressure (MIP), and quality of life (QOL) were determined before and after respective pulmonary rehabilitation course. RESULTS: (1) There were 3, 5, 5 patients in group A, group B, group C dropped off in the course of rehabilitation respectively. (2) The patients' MRC grade after pulmonary rehabilitation in group A and group B decreased compared with that before pulmonary rehabilitation (both P<0.01), but the difference was not significant between two groups (P>0.05). (3) 6MWD, ADL, MEP, MIP of patients in group A and group B increased after pulmonary rehabilitation compared with that before pulmonary rehabilitation, and 6MWD, ADL, MEP, MIP of patients in group A were increased after pulmonary rehabilitation more than those in group B (P<0.05 or P<0.01). (4)The patients' body status, shortness of breath, social activity, home chores in group A and group B, and uneasiness in group A after pulmonary rehabilitation were improved more than those before pulmonary rehabilitation (P<0.05 or P<0.01), but the difference in state of mind, headache, appetite was not markedly different before and after pulmonary rehabilitation in two groups (all P>0.05). The difference in QOL was not marked between group A and group B after pulmonary rehabilitation (all P>0.05). CONCLUSION: (1) The pulmonary rehabilitation with pursed lip respiration and the pulmonary rehabilitation with the guide of respiratory physiology ameliorates dyspnea, improves ADL, QOL, exercise tolerance, function of respiratory muscle in the severe and very severe COPD patients remarkably. (2) The effect of the pulmonary rehabilitation with the guide of respiratory physiology is better than that of the pulmonary rehabilitation with pursed lip respiration, and it can be considered as a more effective pulmonary rehabilitation method for the patients with severe and very severe COPD.

[Changes in serum contents of inflammatory mediators after tetramethylene-disulfo-tetramine poisoning and clinical study of a new treatment regime for the poisoning].

OBJECTIVE: To study the changes in serum contents of beta-endorphin (beta-EP), endothelins (ET), nitric oxide (NO) and tumor necrosis factor (TNF) after acute tetramethylene-disulfo-tetramine (TDT) poisoning and therapeutic effect of a new treatment regime. METHODS: (1) Forty-eight patients with tetramethylene-disulfo-tetramine poisoning (experiment group) were enrolled in this study. The serum levels of beta-EP, ET, NO and TNF were measured upon hospitalization and 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19 days after poisoning, respectively, and compared with those of 30 healthy individuals (control group B). (2) They were treated with the improved regime and compared with patients treated with the conventional regime designated as control group A. RESULTS: (1) In 48 patients treated with improved regime, 45 were cured and 3 died. (2) The serum levels of beta-EP, ET, NO and TNF from 45 patients who were cured were significantly higher at hospitalization compared with those of healthy individuals, with the peak values appeared on day 1 after poisoning in the mild, moderate and severe groups. Beta-EP levels returned to normal range on days 9, 13 and 17 after poisoning respectively in the mild, moderate and severe groups. ET levels returned to normal range on days 7, 13 and 15 after poisoning respectively in the mild, moderate and severe groups. NO levels returned to normal range on days 7, 11 and 11 after poisoning respectively in the mild, moderate and severe groups. TNF levels returned to normal range on days 9, 11 and 17 after poisoning respectively in the mild, moderate and severe groups. (3) The serum levels of beta-EP, ET, NO and TNF in 3 non-survivors were very high at hospitalization and continued to increase in the course of treatment. (4) The cumulative doses of diazepam and Phenobarbital, and the eclampsia time were significantly less in the experiment group than those of control group A. CONCLUSION: (1) The serum levels of beta-EP, ET, NO and TNF are correlated with the severity of tetramethylene-disulfo-tetramine poisoning and general conditions of the patients. (2) When the serum levels of beta-EP, ET, NO and TNF decrease gradually in the course of treatment, prognosis is better. On the contrary, the prognosis is poor when their levels increase gradually. (3) Measures to decrease levels of beta-EP, ET, NO and TNF result in a better prognosis of patients with tetramethylene-disulfo-tetramine poisoning. (4) The improved regime can be considered a better therapeutic strategy in tetramethylene-disulfo-tetramine poisoning.