Objective To produce a hybridoma secreting stable monoclonal antibodies against Aβ22-35 and to develop a detection method for the assay of Aβ. Methods Spleen cells from Balb/cmice immunized with Aβ22-35-KLH were fused with mouse myeloma cells SP2/0. The techniques of immunoprecipitation and western blotting plus ECL were used to investigate the levels of Aβ in the rat brain. Results Two strains of hybridomas (3A8 and 3B2) secreting stable monoclonal antibodies raised against Aβ22-35 were obtained. The subtypes of Aβ22-35 were IgG3. The levels of Aβ in young and older rat brain were 9.8±2.8 and 13.36±2.65 (pmol/12mg brain tissues, x±s), respectively. Conclusion The Aβ22-35 mAb obtained had high titres and specificity. The levels of Aβ in the older rat brain were significantly increased as compared with the young one (P＜0.05).

The combination of Danggui and Honghua (GH) is a popular herb pair commonly used in clinic for the treatment of blood stasis syndrome in China. To evaluate the activating blood circulation and dissipating blood stasis effects of the combination of different proportions of Danggui and Honghua on acute blood stasis rats, and optimize the proportion of GH to have the best activating blood circulation and dissipating blood stasis effect. Acute blood stasis rat model was induced by subcutaneous injection of adrenaline and ice water bath. The blood stasis rats were administrated intragastrically with GH (1 : 0, 4 : 1, 2 : 1, 3 : 2, 1 : 1, 2 : 3, 1: 2, 1 : 4 and 0 : 1) extracts. The whole blood viscosity (WBV), plasma viscosity (PV), and high shear whole blood relative index (HSWBRI), low shear whole blood relative index (LSWBRI), and erythrocyte aggregation index (EAI) were tested to observe the effects of GH on hemorheology of blood stasis rats. And the maximum aggregation induced by adenosine diphosphate (ADP) was tested to observe the effect of GH on platelet aggregation index of blood stasis rats. In addition, the prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT) and plasma fibrinogen (FIB) were tested to observe the effects of GH on blood coagulation function of blood stasis rats. Then principal component analysis and multi-attribute comprehensive index methods were both used to comprehensively evaluate the total activating blood circulation and dissipating blood stasis effects of GH. The results showed that the hemorheological indexes and coagulation parameters of model group both had significant differences with normal group. Compared with model group, GH (1 : 0, 4 : 1, 2: 1, 3 : 2, 1 : 1, 2 : 3, 1 : 2, 1 : 4 and 0 : 1) could improve all the blood hemorheology indexes and regulate part indexes of blood coagulation function and platelet aggregation in acute blood stasis rats. Based on principal component analysis and multi-attribute comprehensive index methods, GH 1 : 1 and GH 3 : 2 both had the best effect of blood circulation and dissipating blood stasis, and the effect of GH 1 : 1 was slightly better than GH 3 : 2. These results suggest that GH could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats. The optimized proportion of GH was consistent with regulations of medicine usage that GH 1 : 1 had the highest frequency used in traditional Chinese formulae. It could provide scientific basis for more effective application of the compatibility between Danggui and Honghua in modern clinic medicine.
Animals ; Blood Coagulation ; Blood Viscosity ; Carthamus tinctorius ; China ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Aggregation ; Hemorheology ; Partial Thromboplastin Time ; Platelet Aggregation ; Principal Component Analysis ; Prothrombin Time ; Rats ; Rats, Sprague-Dawley ; Thrombin Time

Objective To analyze the treatment outcome and related influencing factors of Tibet- an nationality new smear-positive pulmonary tuberculosis patients in Qinghai Province，so as to provide evidence for tuberculosis control and treatment among Tibetan population． Methods Statistical analysis was conducted on 5 564 Tibetan nationality new smear-positive pulmonary tuberculosis cases in Qinghai province who were reported in the China Tuberculosis Information Management System and approved to receive treatment from 2008 to 2017． The main influencing factors were detected by unconditional Logistic regression model analysis，dependent variable was successful treatment or not，independent variables were other factors related to the treatment outcome． Ｒesults The treatment success rate of Tibetan nationality new smear-positive pulmonary tuberculosis cases was 87. 1% ( 4 848 /5 564) ，and the adverse outcome rate was 12. 9% ( 716 /5 564) ． Unconditional Logistic regression model analysis indicated that non-full- course supervision management，living in agricultural and pastoral area，having severe disease，floating population，and age older than 60 years were risk factors of adverse outcome． The odds ratio( OＲ) 95% confidence interval( CI) of the above risk factors were 13. 044( 10. 671-15. 944) ，2. 305( 1. 703-3. 119) ， 2. 090( 1. 346-3. 243) ，1. 967( 1. 443-2. 682) ，and 1. 909( 1. 410-2. 586) ． Clinical consultation，farmers and herdsmen were protective factors． The OＲ( 95% CI) were 0. 451( 0. 375-0. 543) ，and 0. 786( 0. 627- 0. 985) ． Conclusions Treatment success rate of Tibetan nationality new smear positive pulmonary tuberculosis cases was low． Therefore，the directly observed treatment short-course ( DOTS) strategy should be strictly implemented and the full-course supervision management should be strengthened to improve the treatment success rate． More attention should be paid to the elderly，severe，floating，agricultural and pastoral populations among the Tibetan population．

Along with progress of modern science and technology, human is utilizing natural resources and their inherent law more effectively and more efficiently according to their own purposes. Chinese medicine pair (CMP) is relatively fixed combination of two TCMs which was proven to be effective in clinical application. CMP has its inner specification, and it is an intermediate point between single herb and many TCM formulae. With the aid of modern science and technology, and by means of choosing appropriate strategies and approaches, the compatibility rules of CMP might be revealed, which will be significant to develop the compatibility theory of TCM formulae and create modern TCM new drugs.
Animals ; Clinical Trials as Topic ; Drug Interactions ; Humans ; Medicine, Chinese Traditional ; methods ; Research Design ; Systems Biology

OBJECTIVEInsulin-like grouth factor-1 (IGF-1) is polypetide hormone that has demonstrated effects on neural cells. Up to now, there is few reports about the relation between serum IGF-1 and brain damage in neonates with hyperbilirubinemia. This study explored the potential role of serum IGF-1 in neonatal hyperbilirubinemia.

METHODSSerum levels of IGF-1 were measured using ECLIA in 57 term neonates with hyperbilirubinemia and 25 normal term neonates. Meanwhile, total serum bilirubin (TSB), unconjugated bilirubin (USB) and serum albumin (ALB) contents were measured by the automatic biochemistry analyzer and the ratio of USB/ALB (B/A) was calculated. The hyperbilirubinemia group was classified into three subgroups based on serum TSB levels: mild (221-256 micromol/L), moderate (257-342 micromol/L) and severe (>342 micromol/L). Serum TSB levels in the 25 normal neonates were less than 85 micromol/L (control group). NBNA was performed on the day of serum sample collection.

RESULTSSerum IGF-1 levels in the mild, moderate and severe hyperbilirubinemia groups (39.38+/- 8.42, 30.77+/- 4.65 and 26.34+/- 2.05 ng/L, respectively) were obviously lower than those in the control group (50.16+/- 15.73 ng/L) (P< 0.01). There were significant differences among the three hyperbilirubinemia subgroups in serum IGF-1 levels (P< 0.01). Mean NBNA scores in the mild, moderate and severe hyperbilirubinemia groups (35.01+/- 2.26, 32.45+/- 2.74 and 26.77+/- 5.02, respectively) were significantly lower than those in the control group (38.24+/- 0.78) (P< 0.01). Significant differences in the NBNA scores were noted among the three hyperbilirubinemia subgroups (P< 0.01). Serum IGF-1 levels were positively correlated to NBNA scores (r=0.603, P< 0.01) and negatively correlated to the ratio of B/A (r=-0.483, P< 0.01).

CONCLUSIONSSerum IGF-1 levels decreased obviously in neonates with hyperbilirubinemia and correlated to the severity of disease. IGF-1 might be associated with bilirubin-induced brain damage.

Bilirubin ; blood ; Brain ; physiology ; Female ; Humans ; Hyperbilirubinemia, Neonatal ; blood ; complications ; Infant Behavior ; Infant, Newborn ; Insulin-Like Growth Factor I ; analysis ; Male

The metabolic effect of Fo-Shou-San on blood deficiency mice was studied by using metabolomic method. UPLC-QTOF/MS was used to analyze the plasma metabolome in blood deficiency mice. MS data were processed by MarkerLynx software. With multivariate statistical analysis of plasma metabolite profiles, a clear separation among control, blood deficiency model, and Fo-Shou-San groups was achieved. Potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. The metabolic network of blood deficiency was predicted via MetPA database. Twenty-two potential biomarkers were identified and used to explain the thiamine metabolism, arachidonic acid metabolism, sphingolipid metabolism, glyoxylate and dicarboxylate metabolism, histidine metabolism, nicotinate and nicotinamide metabolism, cysteine and methionine metabolism, tryptophan metabolism, starch and sucrose metabolism, tyrosine metabolism and citrate cycle (TCA cycle). Those metabolic pathways were disturbed in blood deficiency mice, but which could be regulated nearly to normal state after Fo-Shou-San administration. In this study, the metabolomics of blood deficiency mice and the action mechanism of nourishing blood effect of Fo-Shou-San were evaluated. The physiological and metabolic state of the organism could be represented comprehensively by using metabolomics. And metabolomics can be used to evaluate the pharmacodynamics and related mechanisms of Chinese medicine and formulae.
Animals ; Arachidonic Acid ; metabolism ; Biomarkers ; blood ; Blood Coagulation Disorders ; blood ; metabolism ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; Female ; Metabolic Networks and Pathways ; drug effects ; Metabolome ; Metabolomics ; Mice ; Mice, Inbred ICR ; Plasma ; metabolism ; Random Allocation ; Spectrometry, Mass, Electrospray Ionization ; Sphingolipids ; metabolism ; Thiamine ; metabolism

OBJECTIVETo assess the effect and safety of nano-Amoni Paste (nmAP) in the treatment of children's anorexia (AR).

METHODSOne hundred and eighty patients of AR were assigned according to the randomized, double-blinded, double-simulated and parallel controlled principle to three groups, the treated group (TG), the positive control group (PCG) and the negative control group (NCG), 60 in each group. The patients in TG were treated by sticking 1.5 ml of nmAP on the acupoint of Shenque (Ren 8) once a day and orally taking placebo liquid 10 ml twice a day; those in PCG and NCG treated with sticking paste of placebo on Ren 8, and oral taking of Shanmai Jianpi Oral Liquid and placebo liquid respectively, 10 ml each time twice per day. The course of treatment for all was 10 days, all patients were treated for 2 courses.

RESULTSThe total effective rate and the effective rate on cardinal symptom in TG was 85.0% and 95.0% respectively, that in PCG 86.2% and 96.55% and in NCG 45.5% and 65.45%, respectively, showing significant difference between groups (P<0.05). Comparison of the clinical manifestation before and after treatment showed significant improvement in volume of food intake, appetite, complexion and reduction of restlessness symptom (P<0.05) in all three groups, and there was no adverse reaction found in them.

CONCLUSIONnmAP is an effective and safe remedy for treatment of Children's anorexia.

Acupuncture Points ; Amomum ; chemistry ; Anorexia ; physiopathology ; therapy ; Appetite ; drug effects ; Body Height ; drug effects ; Body Weight ; drug effects ; Child ; Child, Preschool ; Double-Blind Method ; Eating ; drug effects ; Female ; Humans ; Liposomes ; Male ; Nanoparticles ; Oils, Volatile ; administration & dosage ; adverse effects ; isolation & purification ; therapeutic use ; Ointments ; Phytotherapy ; Plant Extracts ; administration & dosage ; adverse effects ; therapeutic use ; Psychomotor Agitation ; physiopathology ; Treatment Outcome

OBJECTIVETo determine the effect of montelukast, a cysteinyl leukotriene receptor 1 antagonist, on morphological changes in rat neurons after ischemic injury.

METHODSThe in vivo ischemia injury was induced by oxygen-glucose deprivation (OGD) for 2 h and reperfusion (R) for 24 h (OGD/R) in rat neurons primary culture and mixed cortex culture. In the presence or absence of various concentrations of montelukast, neuron number, area of neuron, number of neuritis per neuron, branch number of primary neuritis and primary neurite length were determined for evaluating morphological changes in neurons.

RESULTSOGD/R significantly reduced neuron number, and altered neuron morphology. In cortical neuron cultures, montelukast (0.0001-1 μmol/L) attenuated OGD/R-induced reduction in neuron number, and inhibited OGD/R-induced increase in branch number of primary neuritis. In the mixed cultures, montelukast (0.0001-0.1 μmol/L) increased the primary neurite length, and reduced number of neuritis and branch number of primary neurite after OGD/R.

CONCLUSIONMontelukast has a protective effect on ischemic injury in neurons.

Acetates ; pharmacology ; Animals ; Animals, Newborn ; Cell Hypoxia ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Glucose ; pharmacology ; Leukotriene Antagonists ; pharmacology ; Neurons ; drug effects ; pathology ; Neuroprotective Agents ; pharmacology ; Quinolines ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of cysteinyl leukotriene (CysLT) receptors in the differentiation of rat glioma C6 cells.

METHODSRat glioma C6 cells were treated with the agonist LTD(4), the CysLT(1) receptor antagonist montelukast and the differentiation inducer forskolin. Cell morphology and GFAP protein expression were determined after treatments.

RESULTForskolin (10 μmol/L) induced morphological changes and GFAP protein expression (cell differentiation) in C6 cells, but LTD(4) (0.1-100 nmol/L) did not induce these changes. Montelukast (1 μmol/L) alone did not affect C6 cell differentiation, while it induced the differentiation when combined with the LTD(4) (100 nmol/L).

CONCLUSIONThe CysLT(2) receptor may modulate the differentiation of rat glioma C6 cells.

Acetates ; pharmacology ; Animals ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Colforsin ; pharmacology ; Cysteine ; Glioma ; metabolism ; pathology ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Leukotrienes ; Quinolines ; pharmacology ; Rats ; Receptors, Leukotriene ; agonists

The combination of Angelicae sinensis Radix (Danggui, DG) and Astragali Radix (Huangqi, HQ) is a popular herb pair commonly used in clinic for the treatment of blood deficiency syndrome in China. The aim of this paper is to study the interaction of DG and HQ nourishing and tonifying blood effects by response surface method. The blood deficiency mice were induced by injecting N-acetylphenylhydrazine (sc) and cyclophosphamide (ip). The blood deficiency mice were administrated intragastrically with DG-HQ extracts (0:1, 1: 5, 2:5, 2:3, 1:1, 3:2, 5:2, 5:1, 1:0). The changes of the peripheral blood indexes and organ indexes were observed. The indexes were integrated by comprehensive index method; the interactions of DG and HQ were analyzed by the response surface diagram established with Matlab software. The results showed that DG and HQ at most of their combination ratios had synergic effect. Within the range of 1:5 - 5:1, all of the extracts of DG-HQ showed synergic effect, and among which, high-doses had better effects than low-doses. The highest value (-1) of the synergic effect was showed when DG was 10 - 40 g at the same time of HQ as 90 -180 g, and DG was 50 - 100 g at the same time of HQ as 20 - 100 g. DG-HQ at all combination dosages within Chinese Pharmacopeia (DG: 6 - 12 g, HQ: 9 - 30 g) had certain synergic effect, and Danggui Buxue Decoction (DG: 6 g, HQ: 30 g) also was at this range. The results provided scientific basis to the clinical application of DG and HQ. And the response surface method was firstly applied to quantitatively evaluate the bio-activity change of herb combination, which provided a novel way for modern basic research on the interaction of herbs.
Angelica sinensis ; chemistry ; Animals ; Astragalus membranaceus ; chemistry ; Blood Cell Count ; Drug Combinations ; Drug Interactions ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Mice ; Mice, Inbred ICR ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation