The concentration of 5 nucleosides, uracil, uridine, guanidine, adenine and adenosine in culture of Paecilomyces hepialid was determined by the developed method of HPLC. The HPLC method was performed on a Waters SunFire C18 (4.6 mm x 250 mm, 5 μm) column with methanol-water gradient elution as the mobile phase. The detection wavelength was 260 nm and the colunmn temperature was controlled at 30 °C. The linear range was 10.00-200.00 mg · L(-1) (r = 0.9994) for uracil, 10.10-202.00 mg · L(-1) (r = 0.9992) for uridine, 10.00-200.00 mg · L(-1) (r = 0.9991) for guanidine, 10.30-206.00 mg · L(-1) (r = 0.9992) for adenine and 10.45-209.00 mg · L(-1) (r = 0.9991) for adenosine, respectively. The RSD of precision was 0.032%, 0.035%, 0.039%, 0.049%, 0.00080%, respectively. The average recoveries of uracil, guanidine, adenine, and adenosine were 97.34%, 99.10%, 101.6%, 98.61% and 100.2% with RSD of 1.3%, 2.1%, 0.96%, 0.95%, and 1.3% respectively. The method showed high sensitivity, good selectivity, linearity and repeatability, which was suitable for the content analysis of 5 nucleosides components in P. hepialid and its extracts.
Chromatography, High Pressure Liquid ; methods ; Nucleosides ; analysis ; Paecilomyces ; chemistry

The present study was to investigate the role of dopamine D1 receptors and its relationship with glutamate, N-methyl-D-aspartic acid (NMDA) receptor and α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor in depression induced by chronic unpredictable mild stress (CUMS). CUMS-induced depression model was established in Sprague-Dawley rats, and intrahippocampal microinjections of D1 dopamine receptor agonist SKF38393, non-competitive NMDA receptor antagonist MK-801 and AMPA receptor antagonist NBQX were respectively adopted by rat brain stereotaxic coordinates. The behavioral observations were conducted by measurement of weight changes, sucrose preference test, open-field test and tail suspension test. The concentration of glutamic acid and the expression of its receptors' subunits were detected by HPLC and Western blot, respectively. The results showed that, compared with control group, CUMS rats showed depression-like behavioral changes, higher concentration of glutamic acid, lower expressions of NMDA receptor (NR1) and AMPA receptor (GluR2/3) in hippocampus. Pretreatment with injection of SKF38393 could rescue such depression effect of CUMS, decrease the concentration of glutamic acid, and increase the expressions of NMDA receptor (NR1), AMPA receptor (GluR2/3) in hippocampus. Pretreatment with MK-801 could enhance the antidepressant effect of SKF38393, while NBQX weakened. These results suggest that agonists of D1 dopamine receptor could reduce the concentration of glutamic acid in hippocampus, and its antidepressant effect may be mediated by AMPA receptor partially.
2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine ; pharmacology ; Animals ; Depression ; etiology ; physiopathology ; Dizocilpine Maleate ; pharmacology ; Excitatory Amino Acid Antagonists ; Glutamates ; metabolism ; Hippocampus ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; metabolism ; Receptors, Dopamine D1 ; agonists ; physiology ; Stress, Physiological ; physiology

Objective To observe the combined poisonous effects of fluoride and aluminum on sex hormone of male rats.Methods Sixteen weaned SD healthy male rats aged two week were selected and divided into control group,aluminum group,fluoride group,fluorine-aluminum group,four rats in each group.All rats in the experimental groups were fed with corn collected from the prevailng areas containing different fluorine contents respectively for 90 days.Serum testosterone(T)and estradiol(E2)were detected.Results Compared separatelv with the control group[(3.317±0.635)μg/L],serum T level of fluorine-aluminum group[(15.994±6.558)μg/L]was higher(P＜0.05),but aluminum[(8.134±3.134)μg/L]and fluorine[(1.868±0.367)μg/L]groups had no significant differences(P＞0.05).Compared separately with the control group[(0.319±0.072)nmol/L],E2 level of the fluorine group[(0.172±0.030)nmol/L]being lower(P＜0.05),and it was not significant differences(P＞0.05)in the control group when compared with aluminum group[(0.282±0.012)nmol/L],and fluorine-aluminum group[(0.265±0.047)nmol/L].Fluorine and aluminum interacted with each other(F=9.82,P＜0.05).Conclusion The combined poisonous effects of fluorine and aluminum may influence sex hormone levels of male rats.

AIMTo investigate the effects of sodium sulfonate daidzein (SSD) on stress-induced gastric ulcer and explore its possible mechanism.

METHODSUsing exhausted swimming and counting the number of gastric ulcer to establish the model of stress-induced gastric ulcer. Mouse experience intraperitoneal injection of different doses of SSD and L-NAME, and NDP histochemical method was used to detect the changes of nitric oxide synthase (NOS) positive neurons in stomach.

RESULTSSSD had dose-dependent protective effect on gastric mucosa. L-NAME could prevent stress induced gastric lesion. After combined injection of L-NAME and effective dose of SSD, the protective effect of SSD on gastric mucosa was reinforced. The number of NOS ganglion was constant, and effective dose of SSD had slight effect on NOS-positive neurons in normal mouse while it decreased NOS positive neurons in per area and in per ganglia after stress.

CONCLUSIONThe increased nitric oxide (NO) leads to gastric ulcer during stress, SSD has protective effect on gastric mucosa and this effect may be mediated by inhibiting NOS and restricting the overproduction of NO during stress.

Animals ; Gastric Mucosa ; drug effects ; pathology ; Isoflavones ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Nitric Oxide ; metabolism ; Stomach Ulcer ; Stress, Physiological

Objective To study the results of functional hemispherectomy for pediatric intractable epilepsy with hemisphere lesions , and to explore the indications of functional hemispherectomy , as well as the prognosis. Methods Twenty-four children with intractable epilepsy caused by hemisphere lesions were included , who underwent functional hemispherectomy. All patients were followed up for 3 ~ 48 months. Results All 24 cases obtained improvement of abnormal behavior , and no significant exacerbation of neurological deficits was observed. During the follow-up, 18 cases were in EngelⅠ(75%), 5 in Engel Ⅱ(21%), and 1 in Engel Ⅰ(4%) respectively. No long-term complication was observed. Conclusion Comprehensive presurgical evaluation is a prerequisite for the surgical treatment of pediatric intractable epilepsy caused by hemisphere lesions. Functional hemispherectomy can effectively control seizures and obviously improve the prognosis.

Accumulating evidence indicates an important role of hippocampal dendrite atrophy in the development of depression, while neuropeptide Y (NPY) participates in hippocampal dendrite growth. The present study was aimed to investigate the relationship between NPY and nitric oxide synthase (NOS) in chronic unpredictable mild stress (CUMS)-induced depression. CUMS-induced depression model was established in Sprague-Dawley rats. Intrahippocampal microinjections of NPY, NPY-Y1 receptor antagonist GR231118 and non-specific NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME) were respectively adopted by rat brain stereotaxic coordinates. The behavioral observations were conducted by sucrose consumption test, open field test and forced swimming test. The expressions of NPY, neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) in hippocampus were detected by immunohistochemistry. The results showed that, compared with the control group, rats receiving CUMS for 21 d or intrahippocampal microinjection of GR231118 showed a significant reduction in body weight and depression-like behavior, which included reductions in sucrose preference, locomotor activity, rearing and grooming in open field test, and increased duration of immobility in forced swimming test. Moreover, the expression of NPY significantly decreased (P<0.01), while the expressions of nNOS and iNOS increased obviously in the hippocampal dentate gyrus (DG) and CA3 regions (P<0.01). Intrahippocampal microinjections of NPY prevented the depression-like behavioral changes induced by CUMS and decreased the expressions of nNOS and iNOS in the hippocampal DG and CA3 regions (P<0.01). Intrahippocampal microinjections of GR231118 reduced behavioral ability of the rats dramatically and significantly increased the expressions of hippocampal nNOS and iNOS (P<0.01). Intrahippocampal microinjections of L-NAME suppressed the depression-like behavioral changes induced by CUMS or intrahippocampal microinjection of GR231118. In conclusion, reduced expression of NPY and increased expression of NOS in hippocampus may make significant contributions to CUMS-induced depression. These results suggest that the antidepressant function of NPY associates with down-regulation of NOS expression in hippocampus, possibly mediated via NPY-Y1 receptor.
Animals ; Antidepressive Agents ; pharmacology ; Behavior, Animal ; drug effects ; Depression ; Down-Regulation ; Hippocampus ; drug effects ; Microinjections ; NG-Nitroarginine Methyl Ester ; pharmacology ; Neuropeptide Y ; pharmacology ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Peptides, Cyclic ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Neuropeptide Y ; antagonists & inhibitors

The present study was aimed to investigate the role and relationship between N-methyl-D-aspartic acid (NMDA) receptor and neuropeptide Y (NPY) in depression induced by chronic unpredictable mild stress (CUMS). CUMS-induced depression model was established in Sprague-Dawley rats. Intrahippocampal injections of NMDA, non-competitive NMDA receptor antagonist MK-801 and NPY-Y1 receptor antagonist GR231118 were respectively adopted by rat brain stereotaxic coordinates. The behavioral observations were conducted by sucrose consumption test, open field test and forced swimming test. The expression of NPY in hippocampus was detected by immunohistochemistry. The results showed that compared with the control group, rats receiving CUMS for 21 days or intrahippocampal injection of GR231118 or NMDA showed depression-like behavioral changes, including a reduction in sucrose preference, body weight, locomotor activity, rearing and grooming in open field test, and increased duration of immobility in forced swimming test. Intrahippocampal injection of NMDA decreased the expression of NPY in hippocampal CA3 region and dentate gyrus (DG) region. Intrahippocampal injection of MK-801 improved the depression-like behavioral changes induced by CUMS, and increased the expression of NPY in hippocampal CA3 region and DG region. Co-injection of GR231118 and MK-801showed that GR231118 suppressed the antidepressant effect of MK-801. These data suggest that CUMS might induce depression through excessive release of glutamate (Glu), over-activation of NMDA receptors, and downregulation of NPY. Antidepressant effect of NPY was mainly mediated via NPY-Y1 receptor.
Animals ; Depression ; etiology ; physiopathology ; Dizocilpine Maleate ; pharmacology ; Hippocampus ; metabolism ; physiology ; Male ; Neuropeptide Y ; antagonists & inhibitors ; physiology ; Peptides, Cyclic ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors ; physiology ; Stress, Physiological

AIMTo Investigate the effect of glutamate (Glu) and gamma-aminobutyric acid(GABA) in orbitofrontal cortex (OFC) on regulation of gastric motility.

METHODSUsing microinjection in OFC,together with lesion of related nucleus,and recording the intragastric pressure(IGP).

RESULTS(1) Microinjection of Glu in OFC caused a significant reduce of the amplitude of gastric motility, this effect could be reverse by lesion of amygdala, while lesion of LC had no influence on the effect of Glu. (2) microinjection of GABA in OFC could increase the amplitude of gastric motility significantly,and lesion of LC could abolish this effect,while lesion of amygdala could enhance the effect of GABA more.

CONCLUSIONMicroinjection of Glu in OFC may enhance the normal inhibitory effect of amygdale on gastric motility, and the effect of microinjection of GABA in OFC on gastric motility is closely related with LC.

Amygdala ; physiology ; Animals ; Female ; Frontal Lobe ; metabolism ; physiology ; Gastrointestinal Motility ; drug effects ; Glutamic Acid ; pharmacology ; Locus Coeruleus ; physiology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; pharmacology

Aging can cause degenerative changes in the function of multiple tissues and organs in the body. Gastrointestinal diseases and intestinal dysfunction are very common in the elderly people. The purpose of this study is to explore the effect of the total extract of Astragalus membranaceus (Fisch.) Bge. on intestinal function and gut microbiota homeostasis in natural aging mice, which will provide clues for further mechanism study. The natural aging mice model is established and animal experiments follow the regulations of the Animal Ethics Committee of Nanjing University of Traditional Chinese Medicine. The overall health of the mice was evaluated by the "frailty index" scoring method. The intestinal absorption and transport function were measured by detecting intestinal glucose absorption capacity, transport time, lipase and amylase activities of aging mice. Intestinal inflammation was assessed by detecting inflammatory cytokines by enzyme-linked immunosorbent assay (ELISA). The pathological changes in the intestines of aging mice were tested by hematoxylin-eosin (H&E) staining and alizarin blue (AB) staining. The qRT-PCR method was used to explore the gene transcription level related with the proliferation and differentiation of intestinal stem cells. Microbiota analysis based on 16S rDNA were used to evaluate the composition of gut microbiota. The results showed that Astragalus had a tendency to reduce the "frailty index" of aging mice, but did not show a significant difference. In some indicators of aging phenotype, Astragalus has the most significant effect on hair loss and physical fitness. In terms of intestinal function, Astragalus could increase intestinal glucose absorption capacity, shorten intestinal transportation time and promote lipase secretion in aging mice. The levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-‍α) in the aging intestinal tissue were reduced after Astragalus administration. Astragalus also ameliorated the pathological degeneration of the intestinal tissue of aging mice by increasing the length of small intestinal villi, the thickness of colonic mucosa and goblet cell number. In addition, Astragalus elevated the expression of genes associated with the proliferation and differentiation in jejunum and modulated gut microbiota, especially restoring the abundance of Lachnospiraceae. Taken together, the above research results demonstrate the total extract of Astragalus as a key factor improving the intestinal function and gut microbiota homeostasis of aging mice.

Studies have shown cell-free microRNA (miRNA) circulating in the serum and plasma with specific expression in cancer, indicating the potential of using miRNAs as biomarkers for cancer diagnosis and therapy. This study was to investigate whether plasma miRNA-21 (miR-21) can be used as a biomarker for the early detection of non-small cell lung cancer (NSCLC) and to explore its association with clinicopathologic features and sensitivity to platinum-based chemotherapy. We used real-time RT-PCR to investigate the expression of miR-21 in the plasma of 63 NSCLC patients and 30 healthy controls and correlated the findings with early diagnosis, pathologic parameters, and treatment. Thirty-five patients (stages IIIB and IV) were evaluable for chemotherapeutic responses: 11 had partial response (PR); 24 had stable and progressive disease (SD+ PD). Plasma miR-21 was significantly higher in NSCLC patients than in age- and sex-matched controls (P < 0.001). miR-21 was related to TNM stage (P < 0.001), but not related to age, sex, smoking status, histological classification, lymph node status, and metastasis (all P > 0.05). This marker yielded a receiver operating characteristic (ROC) curve area of 0.775 (95% CI: 0.681- 0.868) with 76.2% sensitivity and 70.0% specificity. Importantly, miR-21 plasma levels in PR samples were several folds lower than that in SD plus PD samples (P = 0.049), and were close to that in healthy controls (P = 0.130). Plasma miR-21 can serve as a circulating tumor biomarker for the early diagnosis of NSCLC and is related to the sensitivity to platinum-base chemotherapy.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; blood ; Carboplatin ; administration & dosage ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; Early Detection of Cancer ; Female ; Humans ; Lung Neoplasms ; blood ; drug therapy ; pathology ; Male ; MicroRNAs ; blood ; Middle Aged ; Neoplasm Staging ; Remission Induction