Ratiometric SERS aptasensing for simultaneous quantitative detection of histamine and tyramine in fishes.

Herein, a SiO2@Ag NPs core/shell nanoparticles were synthesized to fabricate a surface-enhanced Raman spectroscopy (SERS) sensor for the simultaneous determination of histamine (HIS) and tyramine (TYR) based on specific aptamer recognition and ratiometric strategy. SiO2@Ag NPs with 4-thiosaminophenol (4-ATP) and Nile blue A (NBA) molecules were used as an internal standard (IS) and labeled with aptamers corresponding to HIS and TYR, respectively. Raman reporter molecules ROX and Cy5 labeled complementary DNA (cDNA) were then hybridized with aptamers to form rigid double-stranded DNA. After the HIS and TYR were captured by their aptamers, resulting in the dissociation of cDNA and separated from the SERS substrate. Therefore, the SERS signal intensity at 1503 cm-1 of ROX and 1358 cm-1 of Cy5 tagged on the terminal of cDNA decreased with the concentration of HIS and TYR increasing, while the SERS signal intensity at 1079 cm-1 of 4-APT and 592 cm-1 of NBA on the substrate remain stable. Thus, the concentrations of HIS and TYR can be determined by the I1503/I1079 and I1358/I592 values, respectively. This sensing strategy achieves a lower detection limit of 0.2 ng/mL for HIS and 0.05 ng/mL for TYR, respectively, demonstrating promising applications in sensitive detection of BAs in animal-derived foodstuff.

[Bamboo-based medicinal moxibustion for chronic fatigue syndrome: a randomized controlled trial].

OBJECTIVE: To observe the clinical efficacy of bamboo-based medicinal moxibustion for chronic fatigue syndrome (CFS), and to preliminarily explore its action mechanism. METHODS: Sixty-four patients with CFS were randomly divided into a moxibustion group (32 cases, 1 case dropped off, 1 case excluded) and an acupuncture group (32 cases, 2 cases dropped off). The patients in the moxibustion group were treated with bamboo-based medicinal moxibustion, while the patients in the acupuncture group were treated with routine acupuncture. Both groups were treated once a day, 6 days as a course of treatment with 1 day interval, for a total of 2 courses of treatment. Before treatment, 1 and 2 courses into treatment and in the follow-up of 14 days after treatment, the fatigue scale-14 (FS-14) and somatic and psychological health report (SPHERE) scores were observed in the two groups. Before and after treatment, the contents of CD+3, CD+4, CD+8 of peripheral blood T lymphocyte subsets were measured and CD+4/CD+8 ratio was calculated; the clinical efficacy of the two groups was compared. RESULTS: Compared before treatment, the FS-14 and SPHERE scores in the two groups were decreased 1 and 2 courses into treatment and in the follow-up (P<0.01), and the FS-14 and SPHERE scores in the moxibustion group were lower than those in the acupuncture group (P<0.01, P<0.05). Compared before treatment, the contents of CD+3, CD+4 and CD+4/CD+8 ratio in the moxibustion group were increased after treatment (P<0.01). There was no significant difference of CD+3, CD+4, CD+8 and CD+4/CD+8 ratio between before and after treatment in the acupuncture group (P>0.05). After treatment, the contents of CD+3 and CD+4 in the moxibustion group were higher than those in the acupuncture group (P<0.05). The total effective rate was 93.3% (28/30) in the moxibustion group, which was higher than 73.3% (22/30) in the acupuncture group (P<0.05). CONCLUSION: Bamboo-based medicinal moxibustion could improve the physical and mental fatigue symptoms and psychological status in patients with CFS. Its effect may be related to regulating the contents of CD+3, CD+4 of peripheral blood T lymphocyte subsets and CD+4/CD+8 ratio.

[Different stimulation methods on auricular points for primary dysmenorrhea: a randomized controlled trial].

OBJECTIVE: To prove the therapeutic effect of auricular intradermal needling and auricular point sticking on primary dysmenorrhea (PD), and to explore its mechanism. METHODS: A total of 90 patients with PD were randomized into an auricular intradermal needling group, an auricular point sticking group and a placebo group, 30 cases in each one. Neishengzhiqi (TF2), Neifenmi (CO18), Shenmen (TF4), Shen (CO10), Jiaogan (AH6a), Gan (CO12) and Pizhixia (AT4) were selected in the 3 groups, intradermal needling and cowherb seed sticking were applied respectively in the auricular intradermal needling group and the auricular point sticking group, adhesive tape without needle was stuck in the placebo group. Pressing and kneading for 3 to 4 times were required each day, 3 to 4 min each time, and the intervention was started 5 d before menstruation, once every other day, 4 times each menstrual cycle were as one course, and totally 3 courses were required in the 3 groups. The follow-up was adopted at the next menstruation after treatment. The Cox menstrual symptom scale (CMSS) score, the visual analogue scale (VAS) score and the self-rating anxiety scale (SAS) score before treatment, 1,2,3 courses into treatment and at follow-up were compared, the serum levels of PGF2αand PGE2 before and after treatment were detected, and the clinical therapeutic effect was evaluated in the 3 groups. RESULTS: Compared before treatment, the scores of CMSS, VAS and SAS were decreased at each time point of treatment in the auricular intradermal needling group, 2, 3 courses into treatment and at follow-up in the auricular point sticking group and 3 courses into treatment in the placebo group (P<0.001, P<0.05). Compared with the auricular point sticking group, the CMSS scores at each time point of treatment and the VAS scores of 1, 2 courses and at follow-up were decreased in the auricular intradermal needling group (P<0.05). Compared with the placebo group, the CMSS scores were decreased at each time point of treatment in the auricular intradermal needling group and 3 courses into treatment and at follow-up in the auricular point sticking group (P<0.001, P<0.05); the VAS scores were decreased at each time point of treatment in the auricular intradermal needling group and the auricular point sticking group (P<0.001, P<0.05). After treatment, the serum levels of PGF2α were decreased (P<0.05) and the serum levels of PGE2 were increased (P<0.05) in the auricular intradermal needling group and the auricular point sticking group, and the serum levels of PGF2α were lower than the placebo group (P<0.05), the serum levels of PGE2 were higher than the placebo group (P<0.05) in the two groups. Compared with the auricular point sticking group, the serum level of PGF2 was decreased (P<0.05), the serum level of PGE2α was increased in the auricular intradermal needling group (P<0.05). The total effective rates were 93.3% (28/30) in the auricular intradermal needling group and 80.0% (24/30) in the auricular point sticking group, which were both superior to 63.3% (19/30) in the placebo group (P<0.05). CONCLUSION: Auricular intradermal needling and auricular point sticking can both improve the clinical symptom of primary dysmenorrhea, relieve the pain and anxiety, their mechanism may be related to regulating the serum levels of PGF2α and PGE2. The therapeutic effect of auricular intradermal needling is superior to auricular point sticking, and the placebo effect can be preliminarily excluded.

Paraventricular Calcitonin Receptor-Expressing Neurons Modulate Energy Homeostasis in Male Mice.

The paraventricular nucleus of the hypothalamus (PVH) is a heterogeneous collection of neurons that play important roles in modulating feeding and energy expenditure. Abnormal development or ablation of the PVH results in hyperphagic obesity and defects in energy expenditure whereas selective activation of defined PVH neuronal populations can suppress feeding and may promote energy expenditure. Here, we characterize the contribution of calcitonin receptor-expressing PVH neurons (CalcRPVH) to energy balance control. We used Cre-dependent viral tools delivered stereotaxically to the PVH of CalcR2Acre mice to activate, silence, and trace CalcRPVH neurons and determine their contribution to body weight regulation. Immunohistochemistry of fluorescently-labeled CalcRPVH neurons demonstrates that CalcRPVH neurons are largely distinct from several PVH neuronal populations involved in energy homeostasis; these neurons project to regions of the hindbrain that are implicated in energy balance control, including the nucleus of the solitary tract and the parabrachial nucleus. Acute activation of CalcRPVH neurons suppresses feeding without appreciably augmenting energy expenditure, whereas their silencing leads to obesity that may be due in part due to loss of PVH melanocortin-4 receptor signaling. These data show that CalcRPVH neurons are an essential component of energy balance neurocircuitry and their function is important for body weight maintenance. A thorough understanding of the mechanisms by which CalcRPVH neurons modulate energy balance might identify novel therapeutic targets for the treatment and prevention of obesity.

Clinical efficacy evaluation of a traditional Miao technique of crossbow needle therapy in the treatment of knee osteoarthritis: a multi-center randomized controlled trial.

BACKGROUND: Knee osteoarthritis (KOA) seriously reduces quality of life and is a major threat to the health of the middle-aged and elderly. This study aimed to assess the efficacy of Miao crossbow needle therapy vs. acupuncture for KOA therapy. METHODS: This multicenter, randomized controlled trial was performed at three hospitals between April 2016 and December 2016. The patients were randomized to receive crossbow needle (CN) or acupuncture (AT). All treatments were completed within 46 days. Evaluation of treatment was conducted on the 46th, 62nd, and 77th days. The primary endpoint was change of Western Ontario and McMaster Osteoarthritis Index (WOMAC) score on the 46th day. The secondary endpoints included WOMAC score, the Lysholm knee score, the Japanese Orthopedic Association (JOA) knee score, visual analog scale (VAS), and the MOS 36-item short-form health survey (SF-36), on the 46th, 62nd, and 77th day. RESULTS: Finally, data of 301 participants were analyzed for the efficacy of treatment. Compared with AT, there was a larger change of WOMAC score in the CN group after treatment [- 25.0 (95% CI - 27.0, - 23.0) vs. - 18.8 (95% CI - 20.8, - 16.9), P < 0.001]. In the CN group, the WOMAC score was lower at all three time points (P = 0.008, P = 0.003, P < 0.001 respectively), while the Lysholm knee score (P = 0.03) and JOA score (P = 0.013) were higher and the VAS score (P = 0.011) was lower on the 77th day. CONCLUSION: Both Miao crossbow needle therapy and acupuncture reduced the WOMAC score. Miao crossbow needle therapy can be an alternative method for treating patients with knee osteoarthritis. TRIAL REGISTRATION: ChiCTR, ChiCTR-INR-16008032. Registered on 12 March 2016.

Electroacupuncture at ST25 Inhibits Cisapride-Induced Gastric Motility in an Intensity-Dependent Manner.

Background. Previous studies have demonstrated the efficacy of frequency-specific EAS. However, evaluation of intensity-response effects is challenging and has yet to be addressed. Aims. Using cisapride to promote gastric emptying, we measured the intensity-response relationship of EA at ST25 on gastric motility. Methods. We determined the effects of EA at ST25 using intensities (0.5, 1, 3, 5, 7, and 9 mA) on gastric motility in rats injected with cisapride (0.2, 0.02, and 0.002 mg/kg). Results. Utilizing three concentrations of cisapride yielded significantly differing levels of gastric motility. Furthermore, log IC50 values for EAS were different within each group. Given the same EA intensity, cisapride antagonism decreased progressively in each group as a function of drug concentration. The relative amount of cisapride antagonized by EAS did not change in a linear fashion. Finally, EAS at different intensities within the three groups induced a similar pattern of cisapride antagonism. Conclusions. The ability of EAS to elicit a decrease in cisapride-induced gastric motility pressure was demonstrated in this study. The study encompasses construct validity to mirror individualized treatment being based on patients' subjective feelings, not on a set fixed EA intensity. Clinically utilizing EAS at the smallest intensity can achieve the desired therapeutic effect.

Electroacupuncture at ST25 inhibits jejunal motility: Role of sympathetic pathways and TRPV1.

AIM: To investigate whether electroacupuncture (EA) at ST25 affects jejunal motility in vivo and if so, whether a sympathetic pathway is involved. METHODS: Jejunal motility was assessed using a manometric balloon placed in the jejunum approximately about 3-5 cm away from the suspensory ligament of the duodenum in anesthetized animals. The effects of EA at ST25 were measured in male Sprague-Dawley rats, some of which were treated with propranolol or clenbuterol (EA intensities: 1, 3, 5, 7, and 9 mA), and in male transient receptor potential vanilloid-1 (TRPV1) (capsaicin receptor) knockout mice (EA intensities: 1, 2, and 4 mA). RESULTS: Anesthetized rats exhibited three types of fasting jejunal motor patterns (types A, B, and C), and only type C rats responded to EA stimulation. In type C rats, EA at ST25 significantly suppressed the motor activity of the jejunum in an intensity-dependent manner. The inhibitory effect of EA was weakened by propranolol (ß adrenoceptor antagonist) and disappeared with clenbuterol (ß adrenoceptor agonist) induced inhibition of motility, suggesting that the effect of EA on motility is mediated via a sympathetic pathway. Compared with wild-type mice, EA at ST25 was less effective in TRPV1 knockout mice, suggesting that this multi-modal sensor channel participates in the mechanism. CONCLUSION: EA at ST25 was found to inhibit jejunal motility in an intensity-dependent manner, via a mechanism in which sympathetic nerves and TRPV1 receptors play an important role.

Effects of Antioxidants in Human Cancers: Differential Effects on Non-Coding Intronic RNA Expression.

The notion that dietary antioxidants can help fight cancer is popular. However, the mechanism(s) behind the effect of antioxidants in cancer is still unclear. Previous studies indicate that supplements can influence gene expression; however, all of these studies were focused on the coding/exonic gene expression. Studies are now emerging to highlight critical functional roles for RNAs expressed from the non-coding regions. This project was designed to study the effect of antioxidant supplements on non-coding intronic RNA expression in human cancers. Vitamin E, N-Acetyl cysteine (NAC) and Sulforaphane are commonly used supplements to prevent diseases including cancers. We studied the effect of these antioxidant supplements on the non-coding intronic RNA expression using publicly available datasets from a mouse model for lung cancer and prostate cancer cell lines. Although high throughput polyA-enriched RNA-Seq data characterize spliced coding mRNA regions, recent studies reveal the expression of reads from the non-coding intronic regions. Our analyses indicate that cancer cells have higher expression of introns compared to that of normal cells and that treatment with antioxidant supplements reduces the increased expression of introns of several genes. However, we did find high expression of introns of multiple genes including many oncogenes in the supplement treated groups compared to that of the control; this effect was distinct depending on the cell type and the supplement studied. Using RT-PCRs, we validated the expression of introns of two oncogenes, DLK1 and LRG1, known to be key players in lung cancer progression, and demonstrate changed intronic expression with supplement treatment in cancer cells. With regard to the antioxidant system, supplements did not change the intronic RNAs for endogenous antioxidant enzymes except for a significant decrease in the expression of superoxide dismutase (SOD) intronic RNA. Concurrently, we also found that a prolonged (48 h) exposure to Vitamin C, Vitamin E and Green tea extract reduced the enzymatic activity of SOD in lung cancer cells. The results from this study reveal that the antioxidant supplements have a significant effect on the intronic RNA expression of many genes including cancer genes that are not directly linked to the body's antioxidant system. It is important to study this novel effect of antioxidant supplements in detail as it may have a significant role in disease progression.

Preliminary extraction of tannins by 1-butyl-3-methylimidazole bromide and its subsequent removal from Galla chinensis extract using macroporous resins.

In recent years, ionic liquids have become increasingly attractive as 'green solvents' used in the extraction of bioactive compounds from natural plant. However, the separation of ionic liquid from the target compounds was difficult, due to their low vapour pressure and high stabilities. In our study, ionic liquid-based ultrasonic and microwave-assisted extraction was used to obtain the crude tannins, then the macroporous resin adsorption technology was further employed to purify the tannins and remove the ionic liquid from crude extract. The results showed that XDA-6 had higher separation efficiency than other tested resins, and the equilibrium experimental data were well fitted to Langmuir isotherms. Dynamic adsorption and desorption were performed on XDA-6 packed in glass columns to optimise the separation process. The optimum conditions as follows: the ratio of column height to diameter bed was 1:8, flow rate 1 BV/h (bed volume per hour), 85% ethanol was used as eluant while the elution volume was 2 BV. Under the optimised conditions, the adsorption and desoption rate of tannins in XDA-6 were 94.81 and 91.63%, respectively. The content of tannins was increased from 70.24% in Galla chinensis extract to 85.12% with a recovery of 99.06%. The result of ultra-performance liquid chromatography (UPLC)-MS/MS analysis showed that [bmim]Br could be removed from extract.

Assessment of antibacterial properties and the active ingredient of plant extracts and its effect on the performance of crucian carp (Carassius auratus gibelio var. E'erqisi, Bloch).

BACKGROUND: In this study, the antibacterial properties and active ingredient of plant extracts and its effect on the performance of crucian carp (Carassius auratus gibelio var. E'erqisi, Bloch) were assessed. RESULTS: The transmission electron microscopy and flow cytometric analysis showed that the antibacterial activity of plant extracts is due to the disruption of the cell membrane and the leakage of cytoplasmic contents. The UPLC-MS/MS analysis showed that the contents of gallic acid, (-)-epigallocatechin, (+)-catechin, (-)-epigallocatechin gallate, (-)-epicatechin gallate, aloe-emodin, rhein, emodin, chrysophanol, and physcion, were 5.27%, 3.30%, 1.08%, 19.32%, 5.46%, 0.23%, 0.56%, 1.28%, 0.75% and 0.39% in plant extracts, respectively. Results of feeding experiment showed that feeding crucian carp with 1.0% and 2.0% plant extracts significantly enhanced specific growth rate, serum total protein, lysozyme, catalase and superoxide dismutase activities, and decreased the feed conversion rate, malondialdehyde contents and the mortality rate (P < 0.05). CONCLUSION: It can be concluded that plant extracts added to fish feed can act as natural antimicrobial and immunostimulants to prevent pathogenic infection, enhance immune response, and promote growth of the fish.

Detection of melatonin and homocysteine simultaneously in ulcerative colitis.

BACKGROUND: Oxidative stress could be a major contributing factor to the tissue injury that characterize inflammatory bowel disease (IBD). Homocysteine (HCY) could cause oxidative damage to the colon tissue in ulcerative colitis (UC) patients, melatonin (MLT) supplementation could reduce oxidative damage that caused by HCY in vitro and in vivo. In this study, we aimed to determine the levels of plasma HCY and MLT simultaneously in UC patients. METHODS: Collected the clinical data of 112 UC patients and 110 healthy controls (HC). The levels of plasma HCY and MLT were detected by HPLC-FD method. The levels of plasma folate, vitamin B(12) (VitB(12)) were detected by ELISA method. RESULTS: The levels of plasma HCY in UC patients were significantly higher than that in HC (11.27±7.26 µmol/L vs. 8.19±4.81 µmol/L, P=0.000). The levels of plasma MLT in UC patients were significantly lower than that in HC (49.06±31.40 pg/ml vs. 64.28±41.16 pg/ml, P=0.008). The levels of plasma folate and VitB(12) in UC patients were lower than that in HC (7.64±1.95 nmol/L vs. 9.14±1.23 nmol/L, 108.64±32.22 pmol/L vs. 112.64±33.33 pmol/L, P<0.05). The levels of plasma HCY and MLT in UC patients were not correlated with either the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) or the disease activity, localization and duration of UC (P>0.05). The change of increasing levels of plasma MLT but decreasing levels of plasma HCY was shown in UC patients, however, the association between the levels of plasma MLT and HCY were not statistically significant (P>0.05). CONCLUSIONS: The levels of plasma HCY were increased whereas the levels of plasma MLT were decreased in UC patients.

Ionic liquid-based ultrasonic/microwave-assisted extraction combined with UPLC-MS-MS for the determination of tannins in Galla chinensis.

Ionic liquid was used as extraction solvents and applied to the extraction of tannins from Galla chinensis in the simultaneous ultrasonic- and microwave-assisted extraction (UMAE) technique. Several parameters of UMAE were optimised, and the results were compared with of the conventional extraction techniques. Under optimal conditions, the content of tannins was 630.2 ± 12.1 mg g⁻¹. Compared with the conventional heat-reflux extraction, maceration extraction, regular ultrasound- and microwave-assisted extraction, the proposed approach exhibited higher efficiency (11.7-22.0% enhanced) and shorter extraction time (from 6 h to 1 min). The tannins were then identified by ultraperformance liquid chromatography tandem mass spectrometry. This study suggests that ionic liquid-based UMAE is an efficient, rapid, simple and green sample preparation technique.