AIM:To explore whether histamine can regulate the expression of early growth response factor-1 (Egr-1) in the cerebral cortex astrocytes.METHODS:Normal wild-type (WT) mice, histidine decarboxylase knockout ( HDC-KO) mice and histamine treated HDC-KO mice were sacrificed for extracting the total RNA of the cerebral cortex. Primary cultured rat cortical astrocytes were treated with histamine at concentrations of 10 -8 , 10 -7 , 10 -6 , 10 -5 or 10 -4 mol/L for 15, 30, 60, 120 or 240 min.H1 or H2 receptor antagonists were pretreated for 15 min before histamine treat-ment.After histamine treatment, the cell total RNA or protein was extracted.The expression of Egr-1 at mRNA and protein levels was determined by real-time PCR and Western blot.RESULTS:The mRNA level of Egr-1 in cerebral cortex of HDC-KO mice was significantly lower than that in WT mice, while exogenous histamine induced the mRNA expression of Egr-1 in HDC-KO mice.In cultured astrocytes, histamine induced the mRNA expression of Egr-1.The maximum increase in the mRNA level of Egr-1 was produced by histamine at concentration of 10 -5 mol/L.In addition, histamine-induced Egr-1 mRNA accumulation peaked at 30 min after commencing stimulation, while histamine significantly increased Egr-1 protein expression at 60 min.Furthermore, histamine-induced Egr-1 expression was inhibited by H1 receptor antagonist but not H2 receptor antagonist.CONCLUSION:Histamine up-regulates the Egr-1 expression in cerebral cortex and cultured astrocytes, which may attribute to H1 receptor activation.

AIM: To determine the effect of endogenous histamine on transient forebrain ischemia-induced neuronal injury at the late phase of reperfusion using histidine decarboxylase knockout ( HDC-KO) mice.METHODS:Wild-type (WT) and HDC-KO mice were subjected to bilateral common carotid artery occlusion for 30 min followed by 3 d or 15 d of reperfusion.At different time points after reperfusion, the body weight, mortality rate, learning and memory in fear conditioning test and hippocampal CA 1 neuronal density were evaluated .RESULTS: At 1 d after reperfusion , the body weight loss was observed in both WT and HDC-KO mice.At 4 d, 5 d, 6 d and 7 d after reperfusion, the increment in the body weight of the HDC-KO mice was significantly smaller than that of the WT mice .During the period between 8 d and 14 d after reperfusion, the mortality rate of the HDC-KO mice was higher than that of the WT mice (P<0.05).At 14 d after reperfusion , the HDC-KO mice exhibited more aggravated deficits in contextual and cue memory compared with the WT mice.Correspondingly , a more severe CA1 neuronal injury in the HDC-KO mice than that in the WT mice was ob-served at 15 d but not at 3 d after reperfusion (P<0.05).CONCLUSION:Endogenous histamine may attenuate learn-ing/memory deficits and neuronal injury at the late phase of ischemia /reperfusion.However, the involved mechanisms need to be further investigated .

The purpose of this paper is to encapsulate neurotoxin-I (NT-I), a kind of analgesic peptide, into polylactic acid (PLA) nanoparticles (NPs) and to evaluate their transport into the brain after intranasal administration (in) by use of microdialysis sampling technique developed in our laboratory recently. NT-I-NPs (NT-Iradiolabeled with sodium 125I-Iodide) were prepared by a double emulsification solvent evaporation method, and were characterized in terms of surface morphology, particle size distribution, zeta potential and entrapment efficiency. Then, NT-I-NPs were administered intranasally or intravenously to rats and the radioactivities in periaqueductal gray (PAG) were monitored up to 240 min utilizing the microdialysis sampling technique. Nanoparticles prepared were spherical with homogenous size distribution. Their mean particle size and zeta potential measured were (65.3 +/- 10.8) nm and (-28.6 +/- 2.3) mV, respectively. The entrapment efficiency of NT-Iencapsulated into nanoparticles was (35.5 +/- 2.8)%. The brain transport results showed that the time to peak level (Tmax) of NT-I-NPs (in) was (65 +/- 10) min approximately, apparently shorter compared with NT-I-NPs [iv, (95 +/- 10) min] or NT-I [iv, (145 +/- 10) min]. The concentration to peak level (Cmax) and the area under the curves from zero to 4 h (AUC0-4h) of each group followed this order: NT-I-NPs (in) > NT-I-NPs (iv) > NT-I (iv). With nanoparticles as carriers and administered intranasally could be a potential way for centrally active peptides to improve their brain transport. Microdialysis is quite a good technique for the study of drug delivery to the brain.
Administration, Intranasal ; Animals ; Area Under Curve ; Cobra Neurotoxin Proteins ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Lactic Acid ; chemistry ; Male ; Microdialysis ; Nanoparticles ; Particle Size ; Periaqueductal Gray ; metabolism ; Polyesters ; Polymers ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Background and purpose: The incidence of breast cancer increases as patients age, elderly patients account for a large proportion. Due to the insufifcient systemic therapy, more complications and poorly physical condition, the prognosis of elderly patients is often worse than the younger. The aim of this study was to investigate the safety and tolerance with non-anthracyclin regimen as adjuvant chemotherapy in elderly breast cancer patients. Methods:From Nov. 2008 to Jan. 2012, 56 patients (≥65 years) after surgical excision were enrolled into this study. The patients were divided into two groups:TC and PC groups. Each patient received 4 or 6 cycles of chemotherapy of PC (175 and 600 mg/m2, respectively;n=21) or TC (75 and 600 mg/m2, respectively;n=35), administered intravenously every 3 weeks, as adjuvant chemotherapy. Radiation therapy (as indicated) and endocrine therapy, for patients with hormone receptor-positive disease, were administered after completion of chemotherapy. Results: In this study, 50 patients completed chemotherapy as plan, the proportion of two groups were above 90%. After a median follow-up of 33 months, the median disease-free survival(DFS) and overall survival(OS) were not reached. The relapse-free rate and survival rate were 89.5%and 100%in the PC regime group, which were 90.3%and 96.8%in the TC regime group. Major toxicities included:neutropenia, leucopenia, alopecia, nausea, vomiting and various degree of peripheral neuropathy. The incidence of gradeⅢ-Ⅳneutropenia was 76.2%in PC group vs 48.6%in TC group (P=0.044). The most common cause for withdrawing from treatment was to be unable to tolerate the adverse effects. Conclusion:Adjuvant chemotherapy with paclitaxel and cyclophosphamide is safe, tolerable and promising for elderly breast cancer patients.

A simple and quick method is described for the determination of ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in rhizomes of Ligusticum chuanxiong. The 5 active ingredients in the sample was extracted using 40% ethanol and analyzed by reversed-phase high performance liquid chromatography (HPLC). Chromatography separation was performed using Agilent 1100 series HPLC system with a Symmetry C18 column and gradient elution with a mixture of three solvents : solvent A, acetonitrile, solvent B, methanol and solvent C, 1% aqueous acetic acid, 0 min to 5 min A: B: C 20: 40: 40, 5 min to 30 min A: B: C 60 to 100 : 0 : 40 to 0. The effluent was monitored using a VWD detector set at 321 nm (0-4.3 min) and 275 nm (4.31-30 min). The flow rate was set at 1 mL x min(-1) and the injection volume was 10 microL. The column temperature was maintained at 35 degrees C. The calibration curve was linear (r > or = 0.99) over the tested ranges. The average recovery was 94.44%-103.1% (n = 6). The method has been successfully applied to the analysis in different harvest periods of L. chuanxiong samples. In this paper, single-factor randomized block design to study the 5 components content of L. chuanxiong on ten collecting stages. For the L. chuanxiong collected from April 15th to May 30rd, the content of 5 ingredients increased primarily, and then decreased. Determine the appropriate harvest time has important significance to the promotion of the quality of L. chuanxiong.
4-Butyrolactone ; analogs & derivatives ; analysis ; Acetic Acid ; chemistry ; Acetonitriles ; chemistry ; Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Ligusticum ; chemistry ; Methanol ; chemistry ; Solvents ; chemistry ; Time Factors

OBJECTIVETo analyze characterization and determination of nitrogen in the preparation of Qingkailing injection and its intermediate products.

METHODHitich amino acid auto analyzer was used, with the packed analysis column (2.6 mm x 50 mm) and the type of was Hitich 2622 SC resin. The speed of buffer solution and ninhydrin colorimetric solution were 0.4 mL x min(-1) and 0. 3 mL x min(-1) respectively. Program heating was used for controlling column temperature, from 57 degrees C (0.0 min) to 65 degrees C (36 min) to 57 (50 min). The reaction temperature was set at 130 degrees C.

RESULTFree and binding amino acid the existenceare the main form of nitrogen is amino acid in Qingkailing injection and its intermediate products. The total contents of amino acid in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed solution which is made from neutralization of Concha Margaritifera solution extracted by diluted sulfuric acid and Cornu Bubali solution extracted by diluted sodium hydroxide, aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, were 59.56%, 24.88%, 41.84%, 13.49, 14.63% respectively. The type of bonded amino acid was founded in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, and the contents were 9.33%, 15.07%, 16.85%, 19.94% and 19.55%, respectively.

CONCLUSIONThe main resource of the total nitrogen was Bubalus bubalis L. and Isatis indigotica Fort.

Amino Acids ; analysis ; chemistry ; Animals ; Buffaloes ; metabolism ; Chromatography ; methods ; Drugs, Chinese Herbal ; chemistry ; Isatis ; chemistry ; Nitrogen ; analysis ; chemistry

OBJECTIVETo study the chemical constituents of an active fraction (DSS-A-N-30) from Danggui Shaoyao San.

METHODDSS-A-N30 was prepared by macroporous resin chromatography, the compound was isolated by column chromatography on silica gel and RPC-18, the structure was elucidated by spectroscopic methods.

RESULTA new monoterpene glycoside was isolated and identified from DSS-A-N-30.

CONCLUSIONThe new monoterpene glycoside was identified as 4"-hydroxyl-albiflorin.

Bridged-Ring Compounds ; analysis ; isolation & purification ; Chromatography, Gel ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Monoterpenes ; analysis ; isolation & purification

Objective:To investigate the difference between simple posterior interbody fixation and fusion and posterior interbody fixation combined with focus debridement and bone graft fusion for the treatment of mono- and bi-segmental lumbar brucella spondylitis.Methods:A total of 63 patients (42 males and 21 females), aged 50.9±8.18 years (range from 38 to 69 years) with mono- and bi-segmental lumbar brucella spondylitis who received surgical treatment from June 2014 to Feb 2018 were retrospectively analyzed. There were 44 cases of mono-segmental and 19 cases of bi-segmental. Thirty-one cases were treated with single posterior interbody fixation and fusion (PIFF group), and 32 caseswere treated with posterior interbody fixation combined with focus debridement and bone graft fusion (debridement group). The main observation indicators include operation time, intraoperative blood loss, postoperative hospital stay, postoperative medication time, Visual Analogue Scale(VAS), Oswestry Disability Index (ODI), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), Frankel score and clinical efficacy.Results:All of 63 patients were followed up for 27.16±6.07 months (range 15 to 38 months). The operation time of mono-segmental patients of PIFF group was 105.86±16.66 min,the intraoperative blood loss was 295.00±55.11 ml, and the postoperative hospitalization was 4.45±1.53 days, which was significantly shorter than debridement group ( P<0.001), while the postoperative medication time was without significant difference between the two groups ( P>0.05). The opration time of bi-segmental patients of PIFF group was 150.33±26.29 min, the intraoperative blood loss was 242.05±50.56 ml, and the postoperative hospitalization was 4.56±1.50 days, which was significantly shorter than debridement group ( P<0.001), while the postoperative medication time was also without significant difference between the two groups. At the last follow-up time, the VAS scores and ODI values of mono- and bi-segments in PIFF group and debridement group were lower than those preoperation, but there was no significant difference between the two groups ( P>0.05). There was no significant difference in CRP between mono-segments of PIFF group and debridement group at the preoperation, 3 months after operation and the last follow-up time ( P>0.05). The CRP in mono-segments of PIFF group and debridement group decreased at 3 months after the operation compared with that preoperation, and the difference was statistically significant ( P<0.001). There was no significant difference in CRP between bi-segments of PIFF group and debridement group at 3 months after operation and the last follow-up time ( P>0.05). There was no significant difference in ESR between mono- and bi-segments of PIFF group and debridement group at 3 months after operation and the last follow-up time ( P>0.05). There was significant difference in ESR between mono- and bi-segments of PIFF group and debridement group at the preoperation, 3 months after operation and the last follow-up time. There was no statistical difference in the proportion of excellent postoperative clinical efficacy between the two groups. Complications were observed in two patients in PIFF group (6.5%, 2/31) compared with 8 patients in debridement group (25%, 8/32, χ2=4.057, P=0.044). Conclusion:On the basis of standardized anti-brucella drug therapy, simple posterior interbody fixation and fusion for the treatment of brucella spondylitis has a satisfactory surgical effect, and has the advantages of less surgical trauma, shorter time, earlier postoperative movement time and fewer complications.

A field trial was carried out to study the influence of different kinds of spring topdressing on growth, yield and quality of Ligusticum chuanxiong. The results showed that the spring topdressing had effects of improving root length, tiller numbers and plant height to some extent. At the same time the chlorophyll content and dry weight accumulation especially the dry weight of root increased significantly. It also showed that the yield increased and quality was improved significantly. The effect of different treatment with urea58.7 kg x hm(-2)(N 27 kg x hm(-2)) was the best and the treatment with N,P,K the second.
Alkaloids ; metabolism ; Coumaric Acids ; metabolism ; Fertilizers ; Ligusticum ; growth & development ; metabolism ; Seasons

OBJECTIVETo evaluate the diagnostic potential of previously published enterovirus (EV) reverse transcription polymerase chain reaction (RT-PCR) assay in detection of EV in CSF samples from children with a diagnosis of aseptic meningitis and to investigate the clinical characteristics of the patients seen in Shandong.

METHODSEV RNA was detected in 187 CSF samples and serum and/or urine samples of a part of patients by RT-PCR and viral culture technique.

RESULTSRT-PCR was positive in all 62 CSF specimens which were positive by cell culture (100%). In addition, 93 of 125 (74.4%) CSF samples negative by cell culture were RT-PCR positive. In 4 of these 93 (4.3%) patients, viral culture of specimens from other sites (serum or urine) was also positive. The sensitivity of CSF RT-PCR based on clinical diagnosis in patients with meningitis of negative bacterial culture results was 82.9% (155/187), which was considerably higher than the sensitivity of CSF virus culture 33.2% (62/187). The results of RT-PCR can be reported within 4 hours, whereas the viral culture of CSF requires 4.6 days for a cytopathic effect to develop. EV meningitis occurred in a sporadic form and in some areas there were outbreaks. The clinical characteristics of 155 patients with EV meningitis were different in different age groups.

CONCLUSIONEV was one of the most common causes of aseptic meningitis in Shandong area. The RT-PCR assay was rapid, sensitive and specific for the diagnosis of EV meningitis and may be a potential tests to shorten hospital stay and reduce the use of antibiotics.

Central Nervous System Infections ; blood ; diagnosis ; urine ; Child ; Child, Preschool ; China ; Enterovirus ; genetics ; isolation & purification ; Enterovirus Infections ; cerebrospinal fluid ; diagnosis ; Female ; HeLa Cells ; Humans ; Infant ; Infant, Newborn ; Male ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction