Background::Links between alterations in gut microbiota composition and amyotrophic lateral sclerosis (ALS) have previously been reported. This study aimed to examine the microbiota in the nasal cavity of ALS.Methods::Sixty-six ALS patients and 40 healthy caregivers who live in close proximity with patients were enrolled. High throughput metagenomic sequencing of the 16S ribosomal deoxyribonucleic acid (rDNA) gene V3–V4 region of nasal microbiota was used to characterize the alpha and beta diversity and relative abundance of bacterial taxa, predict function, and conduct correlation analysis between specific taxa and clinical features.Results::The nasal microbiome of ALS patients showed lower alpha diversity than that of corresponding healthy family members. Genera Gaiella, Sphingomonas, Polaribacter_1, Lachnospiraceae_NK4A136_group, Klebsiella, and Alistipes were differentially enriched in ALS patients compared to controls. Nasal microbiota composition in ALS patients significantly differed from that in healthy subjects (unweighted UniFrac P = 0.001), while Linear discriminant analysis Effect Size (LEfSe) analysis indicated that Bacteroidetes and Firmicutes dominated healthy nasal communities at the phylum level, whereas Actinobacteria was the predominant phylum and Thermoleophilia was the predominant class in ALS patients. Genus Faecalibacterium and Alistipes were positively correlated with ALS functional rating scale revised (ALSFRS-R; rs = 0.349, P = 0.020 and rs = 0.393, P = 0.008), while Prevotella-9 and Bacteroides operational taxonomic units (OTUs) were positively associated with lung function (FVC) in ALS patients ( rs = 0.304, P = 0.045, and rs = 0.300, P = 0.048, respectively). Prevotella-1 was positively correlated with white blood cell counts (WBC, rs = 0.347, P = 0.021), neutrophil percentage (Neu%, rs = 0.428, P = 0.004), and neutrophil-to-lymphocyte ratio (NLR, rs = 0.411, P = 0.006), but negatively correlated with lymphocyte percentage (Lym%, rs = -0.408, P = 0.006). In contrast, Streptococcus was negatively associated with Neu% ( rs = -0.445, P = 0.003) and NLR ( rs = -0.436, P = 0.003), while positively associated with Lym% ( rs = 0.437, P = 0.003). No significant differences in nasal microbiota richness and evenness were detected among the severe and mild ALS patients. Conclusions::ALS is accompanied by altered nasal microbial community composition and diversity. The findings presented here highlight the need to understand how dysbiosis of nasal microbiota may contribute to the development of ALS.

Human sex glands (ovary and testis) play extremely important roles in reproductive function and sexual behavior by secreting various hormones.However,complex changes in the structure and the function of sex gland occur with ageing,resulting in a series of clinical consequences.This article explores these changes and discuss about their diagnosis and treatment.

Objective To evaluate the feasibility of the lateralization of unilateral medial temporal lobe epilepsy(mTLE)by using arterial-spin-labeling(ASL)based perfusion MR imaging and investigate the changes of perfusion in the regions related to mTLE network and the relationship between the perfusion and the clinical status.Methods Twenty-five patients with left-sided and 23 with right-sided mTLE were enroiled,and 30 healthy volunteers were recruited.The cerebral blood flow(CBF)of related region was measured based on pulsed-ASL sequence on Siemens 3 T scanner.The CBF of the mTLE group were compared with that in the controls by using ANOVA analysis.The asymmetric indices of CBF in the medial temporal lobe were calculated as the lesion side compared with the normal side in matched region in mTLE group.Results Compared with the volunteers,the patients with mTLE showed the decrease of CBF in the bilateral medial and lateral temporal,the frontal and parietal regions relating to the default-mode network and more serious in lesion side.The CBF values of the medial temporal lobe were negatively correlated with the epilepsy duration(r =-0.51,P ＜0.01).The asymmetric index of CBF as-0.01 has a 76.0％(19/25)sensitivity and a 78.3％(18/23)specificity to distinguish the lesion side.Conclusions The decrease of CBF in the temporal and extra-temporal region by ASL-based MRI suggests the functional abnormalities in the network involved by mTLE.The ASL technique is a useful tool for lateralizing the unilateral mTLE.

OBJECTIVETo develop a high performance liquid chromatography coupled with a photodiode array detector (HPLC-DAD) method for simultaneous determination of 7 nucleosides and nucleobases in Fritillaria taipaiensis.

METHODThe analyses were performed on an Agilent Zorbax-SB-Aq-C18 column (4.6 mm x 250 mm, 5 microm) eluted with water and methanol in gradient mode. The flow rate was 1.0 mL x min(-1). The detection wavelength was 260 nm. The temperature of sample manager was set at 25 degrees C, and the injection volume was 20 microL.

RESULTThe investigated compounds including uracil, cytidine, uridine, guanosine, thymidine, adenosine and adenine were shown good linearity (r > or = 0.999 8) over the tested ranges. The average recoveries were within 96.96% - 103.5% with RSD < or = 3.8%.

CONCLUSIONThe accuracy, stability, repeatability and average recovery of the method are satisfying, and the seven nucleosides and nucleobases components in F. taipaiensis can be rapidly and accurately quantified by HPLC-DAD. This work provided helpful information for comprehensive quality evaluation of F. taipaiensis.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fritillaria ; chemistry ; Nucleosides ; analysis ; Reproducibility of Results ; Water ; chemistry

In order to reveal the treatment mechanism of Chinese medicine with the effect of activating blood and resolving putridity, we selected acetyl-11-keto-beta-boswellic acid (AKBA) and arsenic trioxide (ATO), the main monomeric components of frankincense and arsenolite which are two most commonly used Chinese medicine with effect of activating blood and resolving putridity. We combined AKBA and ATO as a compound, and explored its regulatory role in productions and activities of matrix metalloproteinase (MMP)-1, MMP-2 and MMP-9 in human skin fibroblasts (HSFbs) and human acute monocytic leukemia cell line THP-1 in inflammatory state.

Osteoporosis is characterized by reduced bone mass and impaired micro-architectural structure, leading to an increased susceptibility to fractures. It is a complex, multifactorial disorder resulting from genetic factors, environmental factors and gene-environment interactions. Currently there are three opinions on the main pathogenesis of primary osteoporosis in traditional Chinese medicine: kidney deficiency, spleen deficiency, and spleen-kidney deficiency, in which disagreement remains. In this paper, the authors combine the modern etiology of osteoporosis to explain scientific connotation of the three opinions, aiming to comprehend the pathogenesis of primary osteoporosis and strengthen the communication between traditional Chinese medicine and Western medicine, and trying to evaluate the clinical curative effect on osteoporosis.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, which as a group can degrade essentially all extracellular matrix components. The proteolytic property of the MMPs is important during wound healing to remove debris and facilitate cell migration. Targeting towards the decreased MMPs activities is a new treatment strategy for healing chronic wounds. Salvia miltiorrhiza is a popular Chinese herb that could promote chronic ulcers healing for topical use. Salvianolic acid B (Sal B) is the most abundant bioactive component in Salvia miltiorrhiza. The research was designed to explore the inhibitory effects of Sal B on MMP-1, MMP-2 and MMP-9 activities.

AIM: To evaluate the effects of acetyl-11-keto-beta-boswellic acid (AKBA, a main active component from frankincense, one of the traditional Chinese herb for healing wounds) on the activities of matrix metalloproteinase(MMP)-1, MMP-2 and MMP-9.METHODS: Pure human interstitial collagenase (MMP-1) or gelatinase A (MMP-2) was activated by p-aminophenylmercuric acetate (APMA), and was incubated with AKBA for 1 h. The activities of the enzymes were observed by quenched fluorescent substrate. The lysates of rat polymorphonuclear neutrophils [PMNs, rich in gelatinase B (MMP-9)] was incubated with AKBA for 1 h, and activity of MMP-9 was tested by gelatin zymography. Three cell models: activated human dermal fibroblasts by TNF-α, activated THP-1 cells by PMA and fibroblasts-THP-1 co-culture system were established. AKBA was cultured with these cell models for 24 h. The levels of MMP-1, MMP-2 and MMP-9 in the cell culture supernatants were tested by ELISA and activities of MMP-2 and MMP-9 were tested by gelatin zymography assays.RESULTS: AKBA dose-dependently inhibited the activities of human MMP-1 and MMP-2 at the range of 0.1-0.8 mmol/L, with 50% inhibitory concentration (IC50) of 0.18 mmol /L and 0.27 mmol/L, respectively. In the range of 0.05-0.85 mmol/L, AKBA inhibited the MMP-9 activity (P<0.01). Although AKBA promoted fibroblasts to secrete MMP-2, the production of MMP-9 by THP-1 was inhibited. In the cell co-culture system, the inhibitory effects on MMP-1, MMP-2 and MMP-9 productions were also observed.CONCLUSION: AKBA, as a bioactive component of frankincense, has an inhibitory effect on MMPs production and activities, indicating the possible mechanism for healing chronic wounds by frankincense.

OBJECTIVE: To observe the effects of Tribulus terrestris L. saponion (TTLS) on apoptosis in cortical neurons induced by hypoxia-reoxygenation in rats. METHODS: Primary culture of rat cortical neurons was performed in vitro. A model of apoptosis of cortical neurons was established by hypoxia and reoxygenation. Hypoxia for 3 h in neural cells was induced with mixture of 95% N(2) and 5% CO(2), and then reoxygenation in neural cells was induced with mixture of 95% O(2) and 5% CO(2) for 12 h. Different concentrations of TTLS were administered to traditional Chinese herbal medicine-treated group separately during hypoxia and reoxygenation. The apoptosis rate was analyzed quantitatively by flow cytometry with Annexin V-FITC and propidium iodide staining. Mitochondria membrane potential was observed by a confocal laser-scanning microscope with JC-1 fluorescence. Caspase-3/7 activity in cytoplasm was measured by fluorescent plate reader. Bax protein expression was observed by immunohistochemical technique. RESULTS: The percentage of apoptosis was significantly increased, mitochondria membrane potential was obviously decreased, fluorescence of caspase-3/7 activity was increased, and Bax protein was abundantly expressed followed by 3 h of hypoxia and 12 h of reoxygenation (P<0.01). TTLS could inhibit the depression of membrane potential induced by hypoxia and reoxygenation, decrease the activity of caspase-3/7, reduce the expression of Bax protein, and inhibit the apoptosis of the cortical neurons. CONCLUSION: Hypoxia and reoxygenation can induce apoptosis of rat cortical neurons. TTLS can decrease the apoptosis induced by hypoxia and reoxygenation. The mechanism might be related to stabilization of mitochondria membrane potential, inhibition of caspase activity and reduction of Bax protein expression.

AIM: To study the effect of berberine(Ber) on invasion and migration of PG cells from a high metastatic human giant lung carcinoma cell line and to explore its mechanism. METHODS: Agarose drop method was used to detect PG migration; transwell cabin with FN in lower chamber was adopted to detect PG chemotaxis. PG adhesion to FN and martrigel was detected by MTT; PG invasive ability was determined by transwell cabin covered with martrigel. Expression of MMP2/TIMP2 protein and mRNA were detected by quantitative immunocytochemical method and RT - PCR respectively. RESULTS: After PG was treated by Ber( 10 mg/L) for 24 h: 1 ) migration distance of Ber- treated PG cells was markedly shorter than that of control cells (P ＜0. 01 ) and the number of passed membrane cells towards FN was much fewer than that of control cells ( P ＜ 0. 01 ); 2) PG adhesion to FN and martrigel was inhibited remarkably by Ber compared with control PG; 3) the migration of PG cells through the martrigel -coated transwell was significantly inhibited by the addition of Ber; 4) MMP2 expression was reduced significantly(P ＜0. 01 ), while the TIMP2 expression showed up - regulating tendency, but had no differences compared with control group (P ＞ 0. 05). The MMP2/TIMP2 ratio was decreased; 5 )the MMP2 mRNA/TIMP2 mRNA ratio was decreased by Ber. CONCLUSION: Inhibition of cell migration, adhesion to ECM and invasion into ECM of tumor cells and regulation of homeostasis between MMPs and TIMPs to maintain ECM integrity may be the basic mechanism of inhibitive effect of Ber on invasion and metastasis of tumors.