Objective To observe the expression of neural nicotinic acetylcholine receptor subunit α3 (α3nAChR) and extracellular regulated protein kinases (ERK1/2),c-Jun N-terminal kinase (JNK),p38 kinases of mitogen-activated protein kinase (MAPK) pathway in human neuroblastoma cell line SH-SY5Y overexposed to fluoride,and try to investigate the molecular mechanism of cell damage caused by overexposure of fluoride.Methods The SH-SY5Y cell with low expression of α3nAChR suppressed by silence interference RNA served as α3nAChR silence group;the normal SH-SY5Y cell served as control group,and the effect of silencing of αt3nAChR gene in SHSY5Y was detected by Western blotting and real-time PCR;SH-SY5Y cell was treated with different concentrations of fluoride (0.000,0.005,0.050,0.500,1.000,2.500,5.000 mmol/L),the safe concentration of fluoride in SHSY5Y cell was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay;the SH-SY5Y cell of control group and α3nAChR silence group were treated with 4.000 mmol/L fluoride for 0,4,8,12,24,36,48 h according to the results of MTT assay;the expression of ERK1/2,JNK,p38 kinases of MAPK pathway in SH-SY5Y at protein levels was measured by Western blotting.Results The expression of α3nAChR mRNA (0.04 ± 0.03) and protein (12.0 ± 2.5) in α3nAChR silence group was decreased significantly compared with those of control group (1.00 ± 0.11,100.0 ± 11.3,t =24.58,28.80,all P ＜ 0.05).The viability of SH-SY5Y cell treated with 5.000 mmol/L fluoride (0.53 ± 0.15) was decreased significantly compared with that of SH-SY5Y cell treated with 0.000 mmol/L fluoride (1.05 ± 0.05,P ＜ 0.05).The increased expression of phospho-ERK1/2 was found in α3nAChR silence group and control group incubated with fluoride with time prolonged,and the expression of phospho-ERK1/2 increased significantly at time points 24,36 and 48 h (188.33 ± 7.33,200.00 ± 10.01,213.33 ± 11.55;125.33 ± 5.69,136.00 ± 4.52,155.33 ± 6.51) compared to 0 h in the same groups (100.00 ± 0.00,100.00 ± 0.00,all P ＜ 0.05),and the expression of phospho-ERK1/2 was higher significantly in α3nAChR silence group than those of control group (t =9.26,7.63,5.72,all P ＜ 0.05);no change of expression of total-ERK1/2 in the two groups was found with the passage of time.The gradually increased expression of phospho-JNK was found in α3nAChR silence group and control group,among which,the expression of phospho-JNK in o3nAChR silence group at time points 12,24,36 and 48 h (154.00 ± 6.25,149.00 ± 5.57,156.00 ± 6.08,141.67 ± 2.52) and in control group at 8,12,24,36,48 h (133.33 ± 10.69,173.00 ± 4.00,175.00 ± 11.79,200.67 ± 11.93,200.33 ± 18.58) was compared to those at 0 h in the same groups (100.00 ± 0.00,100.00 ± 0.00),and the difference was significant (all P ＜ 0.05);the higher expression of phospho-JNK was found in α3nAChR silence group other than control group at 8,12,24,36,48 h (t =-4.28,-5.02,-2.89,-8.33,-6.18,all P ＜ 0.05);no change of expression of total-JNK was found in the two groups (P ＞ 0.05).The increased expression of phospho-p38 was detected in control group at time points 24,36 and 48 h (120.33 ± 4.51,122.00 ± 7.55,119.67 ± 7.57) compared to 0 h in the same groups (100.00 ± 0.00,all P ＜ 0.05),and the expression of phospho-p38 was significantly higher than that in α3nAChR silence group at the same time points (93.33 ± 9.61,94.00 ± 5.01,98.33 ± 5.69,t =-4.01,-6.73,-5.59,all P ＜ 0.05);no change of expression of total-p38 was found in the two types of SH-SY5Y cells treated with fluoride (P ＞ 0.05).Conclusion When SH-SY5Y cells are exposed to fluoride;activation of ERK1/2 may be not depend on α3nAChR;α3nAChR may have protected the cell from apoptotic injury caused by activation of JNK pathway,and the activation of p38 may be depend on nAChRα3.

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 observe the effect of Yi Jin Jing (Sinew-transforming Qigong Exercises) on the muscle strength in senile sarcopenia. Methods: Sixty-five old people with sarcopenia were randomized into Yi Jin Jing group and a blank control group. Thirty-three patients in Yi Jin Jing group practiced Yi Jin Jing (Sinew-transforming Qigong Exercises), while 32 patients in the blank control group didn't receive any interventions. The muscle strength was measured before and after 12-week training. Results: During the study, each group had 1 dropout. The muscle strength was improved after 12-week training in Yi Jin Jing group, and the difference was statistically significant (P<0.05); there was no significant difference in the blank control group (P>0.05). After the intervention, there was a significant difference between Yi Jin Jing group and the blank control group in comparing the muscle strength (P<0.05). Conclusion: Constant Yi Jin Jing (Sinew-transforming Qigong Exercises) training can notably improve skeletal muscle strength in senile sarcopenia.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

To clarify the prognostic contribution of peripheral blood cell and bone marrow megakaryocyte counts in patients with idiopathic thrombocytopenic purpura, a series of data of 299 ITP patients including the counts of peripheral white blood cells and platelets, their increase potentials after treatment and the megakaryocytic counts on the bone marrow smears at diagnosis as well were collected and retrospectively analyzed to correlate with the disease development. The results showed that peripheral white blood cell and platelet counts at diagnosis were not associated with the prognosis, but positively associated with the increment of platelet counts after treatment. The cure rate reached up to 94.9% in the group with the platelet level restored to 100 x 10(9)/L in two weeks of therapy. The numerous megakaryocyte counts in bone marrow at diagnosis indicated good prognosis that the cure rate was up to 86.1% when the counts were more than 100 per 1.5 x 3 cm of smear. In conclusion, bone marrow examination on the quantity and quality of megakaryocyte would be critical for diagnosis and evaluation of the prognosis. Consecutive platelet counts during therapy is useful to estimate the disease development.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; cytology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukocyte Count ; Male ; Megakaryocytes ; cytology ; Middle Aged ; Platelet Count ; Prognosis ; Purpura, Thrombocytopenic, Idiopathic ; blood ; therapy

This device is used to measure the tension and the strength of a experimental healed wound in the research on stimulating the wound healing using vitamin A and vitamin C.The effect of vitamin A and vitamin C to the wound healing degree can be distinguished by using this device.

Objective To investigate the alexithymia, mental health condition and their relationship in female rheumatoid arthritis patients. Methods Fifty-four female patients with rheumatoid arthritis and 50 healthy women were enrolled and assessed with Chinese version of the 20-item Toronto Alexithymia Scale (TAS-20) and the symptom checklist-90(SCL-90). Disease duration and DAS28 of the patients were also recorded and calculated. The data were statistical analyzed by t test between the two groups and other block groups. The association between TAS-20, SCL-90, duration and DAS28 was assessed using Spearman corre-lations. Results ① The TAS-20 total score and Tf1, Tf2, Tf3 factors score of female rheumatoid arthritis patients were significantly higher than those of the control group (57±9, 51±7, t=4.15, P<0.01); it was also the same with the total score and factors score of SCL-90 (beside the paranoid factor) between patients and control groups (165±50, 138±41, t=3.06, P<0.05). ② Correlation analysis showed significantly positive correlation between Tf1 factor and all factors of SCL-90, also between Tf3 factor and obsession-compulsion, interpersonal sensitivity, phobias and paranoid factors(P<0.05). ③ Only hostility factor was different between the higher TAS-20 total score group and the lower one. However, the obsession-compulsion, anxiety and phobias factors scores of the higher Tf1 score group were significantly higher than those of the lower one (P<0.05); The higher Tf3 score group was significantly different from the lower one in obsession-compulsion, interpersonal sensitivity, depression, hostility, paranoid and psychoticism factors (P<0.05). The Tf1 factor score of higher SCL-90 total score group was significant higher than that of the lower one (P<0.05). ④ There were significantly positive correlation between duration and depression and paranoid factors, also between
DAS28 and anxiety and paranoid factors (P <0.05). Conclusion There is obvious alexithymia and psychological problems in female rheumatoid arthritis patients; the Tf1 and Tf3 factors of TAS-20, duration and DAS28 are all major factors related to the psychological state.

Objective To examine the effect of rapid infusion test guided by extravascular lung water index (EVLWI) on hemodynamics in critically ill patients at different states in order to guide volume resuscitation.Methods A prospective observation was conducted.Forty critically ill patients admitted to Department of Critical Care Medicine of Tianjin Third Central Hospital from June 2012 to April 2014 were enrolled.Based on the levels of EVLWI and pulmonary vascular permeability index (PVPI) and the cardiac function,the patients were divided into four groups:septic patients with normal EVLWI and PVPI (n =17),septic patients with increased EVLWI and PVPI (n =3),septic patients with increased EVLWI and normal PVPI (n =4),and coronary heart disease and heart failure patients with normal EVLWI and PVPI (n =16).The rapid infusion test was conducted in all patients using lactated Ringer solution 250 mL,followed by infusion of crystalloid with rate of 150 mL/h.The conditions of mechanical ventilation and vasoactive drugs were not changed during study.The changes in EVLWI,intrathoracic blood volume index (ITBVI),and cardiac index (CI) before capacity load,at immediate capacity load,and 15,45,105 minutes after load were determined by pulse indicator continuous cardiac output (PiCCO).On the base of volume status before and after the liquid infusion,the standard for the changes were:stroke volume (SV) increased by 12％-15％,central venous pressure (CVP) greater ≥ 2 mmHg (1 mmHg =0.133 kPa),CI ＞ 15％,and ITBVI change greater than 10％.Results There were no statistically significant differences in the observed indicators at the each time point before and after rapid infusion test among the four groups (all P ＞ 0.05).In septic patients with normal EVLWI and PVPI group,ITBVI was slightly increased by 5.4％-9.7％ from 15 minutes to 45 minutes after rapid infusion test.In coronary heart disease and heart failure patients with normal EVLWI and PVPI group,the EVLWI was increased by 11.9％,5.9％,and 14.7％ respectirely at 15,45,and 105 minutes,ITBVI was slightly increased by 6.4％ at 45 minutes,CI was increased by 29.5％ immediately after rapid infusion.In septic patients with increased EVLWI and PVPI group,CVP was increased by 8 mmHg immediately,EVLWI was increased significantly by 15.8％ at 45 minutes,ITBVI was slightly decreased by 10.0％ at 45 minutes,CI was increased by 24.7％ immediately,and increased by 17.0％ at 105 minutes,and PVPI was increased by 15.6％-28.1％ at 15-105 minutes after rapid infusion.In septic patients with increased EVLWI and normal PVPI group,CVP was increased by 1.5 mmHg at 15 minutes,EVLWI was increased immediately,which was increased by 17.4％,24.0％,and 31.4％ respectively at 15,45,and 105 minutes,ITBVI was increased by 13.9％ at 15 minutes,CI was increased by 16.1％ at 15 minutes after rapid fluid infusion.Conclusions Rapid fluid replacement in critically ill patients with crystalloid,regardless of whether the EVLWI was normal or increased,the short-term response was affected by the volume and cardiac function of patients.Different status of patients showed different volume effect curve:no significant changes in hemodynamic parameters were found in patients with normal EVLWI and volume parameters.In patients with potential cardiac dysfunction,CI and EVLWI increased significantly; regardless of PVPI increased or normal,EVLWI and CI were increased in patients with elevated EVLWI; two different changes could be found in the two types of pulmonary edema while ITBVI was increased.

Objective To investigate the effects of subchronic arsenic exposure on caspase-3 expression in Leydig cells and serum testosterone level in rats. Methods Forty SD rats were divided into four groups based on body mass (160-220 g) by random number table, 10 in each group and treated with As2O3 through drinking water at the doses of 0.0 (distilled water), 2.0, 12.0 and 60.0 mg/L for 14 weeks. Blood sample from heart was collected, serum testosterone was measured by enzyme linked immunosorbent assay (ELISA); the testicular tissue of rats was taken, the expression of caspase-3 was assayed by immunohistochemical staining, and its average gray value was analyzed with image analysis software (Biomias). Results There were statistically significant differences of the serum testosterone levels between groups (F= 37.99, all P< 0.05). Compared with the control group [(3.082 3 ± 0.653 0) ng/L], serum testosterone levels in middle-dose and high-dose groups [(1.694 6 ± 0.255 4), (1.350 5 ± 0.281 9)ng/L] were significantly lower (all P< 0.05). The numbers of positive cells of caspase-3 were 8.10 ± 1.91, 9.80 ± 2.15,10.00 ± 1.83 and 10.90 ± 2.38 in each vision in the 4 groups, there were statistically significant differences between groups (F=3.17, all P<0.05), compared with the control group , the middle-dose and high-dose groups were significantly higher (all P<0.05);the average gray values of caspase-3 were 135.10 ± 6.89, 130.00 ± 3.30, 117.10 ± 4.28, and 113.10 ± 5.38, there were statistically significant differences between groups (F = 41.09, P < 0.05), compared with the control group, the middle-and high-dose groups were decreased (all P<0.05). Conclusions One possible mechanism of As2O3 on male germ cell toxicity may be through up-regulation of the expression of caspase-3 in Leydig cells of SD rats and initiating the pathway of the apoptosis signal, which can lead to increased apoptosis of Leydig cells, decreased concentrations of testosterone, and damaged reproductive function in rats.

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.