The present study aims to predict the action targets of antidepressant active ingredients of Xiaoyaosan to understand the "multi-components, multi-targets and multi-pathways" mechanism. Using network pharmacology, the reported antidepressant active ingredients in Xiaoyaosan (saikosaponin A, saikosaponin C, saikosaponin D, ferulic acid, Z-ligustilide, atractylenolide I, atractylenolide II, atractylenolide III, paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid and pachymic acid), were used to predict the targets of main active ingredients of Xiaoyaosan according to reversed pharmacophore matching method. The prediction was made via screening of the antidepressive drug targets approved by FDA in the DrugBank database and annotating the information of targets with the aid of MAS 3.0 biological molecular function software. The Cytoscape software was used to construct the Xiaoyaosan ingredients-targets-pathways network. The network analysis indicates that the active ingredients in Xiaoyaosan involve 25 targets in the energy metabolism-immune-signal transmutation relevant biological processes. The antidepressant effect of Xiaoyaosan reflects the features of traditional Chinese medicine in multi-components, multi-targets and multi-pathways. This research provides a scientific basis for elucidation of the antidepressant pharmacological mechanism of Xiaoyaosan.
Antidepressive Agents ; pharmacology ; Benzoates ; Bridged-Ring Compounds ; Coumaric Acids ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; Glucosides ; Glycyrrhizic Acid ; Lactones ; Medicine, Chinese Traditional ; Monoterpenes ; Sesquiterpenes ; Software

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

OBJECTIVETo investigate the effect of plasma neuropeptide Y (NPY), neurotensin (NT) and their relationships on the diurnal rhythm of BP and target organ damage for essential hypertension.

METHODSAmbulatory BP monitoring (ABPM) for 24 hours was performed in ninety patients with essential hypertension and thirty healthy subjects. The patients were divided into two groups: 55 dippers and 35 non-dippers according to the ABPM results. Plasma NPY and NT levels in subjects were measured at 8:00, 16:00 and 2:00 o'clock by using radioimmunoassay method. Target organ functions were measured.

RESULTSThe plasma NPY level was higher and NT was lower in patients with hypertension those in normal controls (P < 0.001). Compared with dippers, non-dippers had a higher NPY and a lower NT plasma levels, especially appearing at 2:00 o'clock. The patients with left ventricular hypertrophy, stroke or kidney damage had a higher NPY and a lower NT levels compares with those without target organ damage.

CONCLUSIONThe changes in plasma NPY and NT may contribute to disturbance of the diurnal rhythm of blood pressure in patients with essential hypertension, especially in those with target organ damages, which may serve as a pathophysiologic mechanism for target organ damages in hypertension.

Adult ; Aged ; Blood Pressure ; Blood Pressure Monitoring, Ambulatory ; Circadian Rhythm ; Female ; Humans ; Hypertension ; blood ; physiopathology ; Male ; Middle Aged ; Neuropeptide Y ; blood ; Neurotensin ; blood

Objective To investigate the spiral CT features of gastric carcinoma in the invasion and metastasis and its correlation with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast grouth factor(bFGF).Methods Spiral CT plain scan and triphasic enhanced scans were performed in 83 patients.The postoperative specimens were embedded with paraffin to obtain 5?m thickness tissues and stained with HE and immunohistochemistry.Spiral CT findings were compared with the expression of phosphatase and tensin horology deleted on chormosome ten(PTEN)and basic fibroblast growth factor(bFGF).Results(1)The accuracy of spiral CT in T and N staging of gastric carcinoma was 94.0%(78/83)and 89.2%(74/83),respectively.(2)The expression of PTEN was 47.0%(39/83)in gastric carcinoma.The expression of PTEN in T_(3.4)(40.8%,29/71)and N_(1+2) (38.3%,23/60)gastric carcinoma was significantly lower than that of T_2(10/12)and N_0(16/23)gastric carcinoma,respectively(X~2=7.439,P=0.006;X~2=6.511,P=0.011).(3)The expression of bFGF was 63.9%(53/83)in gastric carcinoma.The expression of bFGF in T(3.4)(70.4%,50/71)and N_(1+2) (71.7%,43/60)gastric carcinoma was significantly higher than that of T_2(3/12)and N_0(10/23)gastric carcinoma,respectively(X~2=7.314,P=0.007;X~2=5.724,P=0.017).(4)Both PTEN-positive expression and bFGF-positive expression were detected in 16 specimens.The expression of PTEN(41.0%, 16/39)was negatively correlated with that of bFGF(30.2%,16/53)(r=-0.447,P=0.000). Conclusion Spiral CT triphasic enhanced scans combined with biologic characteristics can improve diagnostic accuracy of gastric carcinoma in the invasion,metastasis and prognosis.

Adult ; Embolism, Amniotic Fluid ; etiology ; therapy ; Female ; Humans ; Pregnancy

Objective To compare the efficacy of different target concentrations of sufentanil target-controlled infusion used to supplement topical anesthesia for fiber-optic bronchoscopy (FOB)-assisted awake nasotracheal intubation in patients with obstructive sleep apnea syndrome (OSAS).Methods Forty-five ASA physical status Ⅱ or Ⅲ patients with OSAS,aged 28-60 yr,with body mass index of 30-40 kg/m2,scheduled for elective surgery,were randomly assigned into 3 groups (n =15 each):control group (group C) and sufentanil with the target plasma concentration of 0.4 ng/ml (group S1) and 0.6 ng/ml groups (group S2).Naso-pharyngeal and laryngeal mucous membrane was sprayed with 2％ lidocaine mixed with 1％ ephedrine for topical anesthesia in both groups.In addition 1％ tetracaine 3 ml was injected into trachea through cricothyroid membrane.FOB-assisted awake nasotracheal intubation was performed after the target concentration was achieved.The degree of airway obstruction was scored during intubation.The highest values of MAP and HR,rate-pressure product ＞ 12 000,decreased respiratory rate and hyoxemia were recorded during the period between induction of anesthesia and 3 min after intubation was completed.The changes in MAP and HR as percent of baseline values were calculated.Before topical anesthesia (T0),when target concentrations were reached (T1),and at 1 and 3 min after intubation (T2,3),blood samples were taken to determine the plasma concentrations of epinephrine (E),norepinephrine (NE) and cortisol.Results Compared with group C,the airway obstruction score was significantly decreased in group S1,the incidence of changes in MAP and HR ＞ 30％ of baseline values and rate-pressure product ＞ 12 000 was decreased,the plasma concentrations of E,NE and cortisol were decreased in S1 and S2 groups,and the incidence of the respiratory rate was decreased and hypoxemia was increased in group S2 (P ＜ 0.05).Compared with group S1,the airway obstruction score were significantly decreased,and the incidence of respiratory rate was decreased and hypoxemia was increased in group S2 (P ＜ 0.05).Compared with the baseline value at T0,the plasma concentrations of E,NE and cortisol were significantly increased at T2,3 in group C,while decreased at T1 in S1 and S2 groups (P ＜ 0.05).Conclusion Compared with pure topical anesthesia,sufentanil with the target plasma concentration of 0.4 ng/ml does not induce respiratory depression,maintains hemodynamics stable,attenuates the stress responses and provides better intubation conditions when used to supplement topical anesthesia for FOB-assisted awake nasotracheal intubation in patients with OSAS.

Objective To investigate the effect of different doses of sufentanil combined with dexmedetomidine ( DEX) on hemodynamic and Narcotrend index ( NI) during pediatric anesthesia induction. Methods A total of 45 children with lower abdominal surgery were randomly divided into three groups evenly: sufentanil 0. 1 μg·kg-1+ DEX (S1 group),sufentanil 0. 2 μg·kg-1+DEX (S2 group),and sufentanil 0. 3μg·kg-1+DEX (S3 group). Patients in each group began with intubation at the peak point of administration. Blood pressure,heart rate,perfusion index (PI) and NI were detected at the baseline (t0), delivering DEX 0.5 μg·kg-1·h-1 and sufentanil intravenously for 5 min (t1),delivering sufentanil for 3 min (t2),time of intubation ( t3 ) ,1 min ( t4 ) ,and 5 min ( t5 ) after intubation. The application rate of atropine and propofol was recorded. Patient recovery time and adverse reactions were observed. Results Compared with basicline value at t0 time point, hemodynamic parameters and NI were decreased at t1 and t2 ,while PI was increased in both groups. At t3 ,t4 ,and t5 ,all of the indicators in S1 group were significantly different from those at t0 ,and also significantly different from those in S2 and S3 group. Six patients were treated with propofol in S1 group and four presented with agitation after operation,more than S2 and S3 groups. Three patients were treatment with atropine in S3 group. Conclusion Sufentanil (0. 2 μg·kg-1 ) combined with dexmedetomidine can be used to induce intubation for pediatric anesthesia with stable hemodynamic profile and low incidence of adverse effects.

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

Objective To investigate the expression of extraeellular signal-regulated protein kinase (ERK1/2)pathway in rat brains with fluorosis and the effects of fluoride on learning and memory of the rats,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of the ion.Methods Seventy-two SD rats were divided into 3 groups and 24 rats were in each group.Three groups were fed respectively with different concentrations of fluoride(NaF)for 6 months to establish rat models with fluorosis.Controls were fed with tap water (NaF＜0.5 mg/L):lower and higher concentration group were fed with water containing NaF(5,50 ms/L).Animals are sacrificed after 6 months of treatment with fluoride and the dissected brains were kept for analysis.The protein levels of ERK1/2 in rat brains were detected by Western-blotting and the mRNA level by RT-PCR. The spatial learning and memorizing ability was measured by Morris water maze test. Results The ERK1/2 protein in control group,lower and higher concentration group was 0.944±0.10,1.253±0.02,1.953±0.07,the differece being statistieally sighificant between any two groups (P ＜ 0.05). The phospho-ERKl/2 protein in control group,lower and higher concentration group was 0.73±0.08,0.77±0.07,1.28±0.11,the differece being statistieally sighificant between any two groups(P ＜ 0.05);the activation rate of phospho-ERK1/2 in lower and higher concentration group [(68.4± 3.8)%,(64.1±3.2)%] was decreased compared to control group[ (82.3±10.7)%],the differece being significant(P ＜ 0.05). In the navigation trial,longer escape latencies of lower concentration group on the second, the third,the fifth and the sixth day were observed[ (46.0±8.0),(24.0±2.7),(8.9±5.3),(7.4±4.1 )s] compared to the control[ (39.3±6.9),(19.1±9.1 ),(8.3±3.4),(4.8±2.7)s],the differece being significant (P ＜ 0.05 or ＜ 0.01 );the similar results were also observed in the higher concentration group[ (36.9±16.8),(37.7±12.9), (19.7±7.6),(12.2±5.7 )s],and the escape latencies of the higher concentration group on the third,the fifth and the sixth day were longer than that in lower concentration group. In the probe test,the rats took more time to reach the first cross in lower and higher concentration group[(1.17±0.75),(4.18±1.10)s] than control group[ (5.89± 0.56 ) s ],the differece being significant (P ＜ 0.05 or ＜ 0.01 ) ;stayed shorter [ ( 17.05±4.25 ),(18.20±4.57 ) s ] than control [(25.37±5.65 )s ] in platform area (P ＜ 0.01 );the activation rates of ERK1/2 were directly correlated with the time taken to reach the first cross platform located in the probe test(r = 0.364,P ＜ 0.05) and the activation rates were also directly correlated with the escape latencies on the sixth day(r = 0.497,P ＜ 0.05). Conclusion Long-term exposure of excessive fluoride induces the change of expression and activating rate of the ERK1/2 in rat brains,leading to the decreased capacity of learning and memory.

Objective To investigate the changes of oxidative stress level in brain tissues and serum, and learning and memory in rats with oxidative stress level in nerve damage in chronic fluorosis. Methods The rats were randomly divided into 3 groups according to the body weight, eight rats in each group, i.e., control group, drinking water containing less than 0.5 mg/L of fluoride; lower fluoride exposure group, drinking water containing 5 mg/L of fluoride; higher fluoride exposure group, drinking water containing 50 mg/L of fluoride. The animals were examined six months after initiating the experiment. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA), as well as learning and memory, were measured. Results Escape latency in higher fluoride exposed group[ (14.37±3.48)s] was significantly higher than that of controls[ (5.84±1.87)s] and exposed te lower fluoride [ (7.18±1.42)s], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.17±0.11)× 103 U/L , (0.79±0.11)×103 U/g Pr] ,the rats exposed to higher fluoride and lower fluoride exhibited lower levels of T-AOC [(1.37±0.27)×103 U/L,(0.24±0.06)×103 U/g Prand (1.20±0.14) x 103 U/L,(0.41 ~ 0.10)×103 U/g Pr], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.34±0.16) mmoL/L, (2.97±0.11)mmol/g Pr] and low fluoride exposed group[ (2.68±0.33)mmoL/L, (3.38±0.21)mmol/g Pr], higher level of MDA were observed in higher fluoride exposed group[ (3.72±0.59)retool/L, (4.01±0.21)mmol/g Pr], the difference being statistically significant(P<0.05). Conclusion The results indicated that higher amount of fluoride induced an increased level of oxidation, which might result in the decreased capacity of intelligence of rats with fluorosis.