This study aims to explore the neuroprotective effect of bilobalide(BB) and the mechanisms such as inhibiting inflammatory response in macrophage/microglia, promoting neurotrophic factor secretion, and interfering with the activation and differentiation of peripheral CD4~+ T cells. BB of different concentration(12.5, 25, 50, 100 μg·mL~(-1)) was used to treat the RAW264.7 and BV2 cells for 24 h. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and cell counting kit-8(CCK-8) were employed to detect the cytotoxicity of BB and appropriate concentration was selected for further experiment. Lipopolysaccharide(LPS) was applied to elicit inflammation in RAW264.7 and BV2 cells, mouse bone marrow-derived macrophages(BMDMs), and primary microglia, respectively. The effect of BB on cell proliferation and secretion of inflammatory cytokines and neurotrophic factors was detected by enzyme-linked immunosorbent assay(ELISA). Spleen monocytes of C57BL/6 female mice(7-8 weeks old) were isolated, and CD4~+ T cells were separated by magnetic beads under sterile conditions. Th17 cells were induced by CD3/CD28 and the conditioned medium for eliciting the inflammation in BMDMs. The content of IL-17 cytokines in the supernatant was detected by ELISA to determine the effect on the activation and differentiation of CD4~+ T cells. In addition, PC12 cells were incubated with the conditioned medium for eliciting inflammation in BMDMs and primary microglia and the count and morphology of cells were observed. The cytoto-xicity was determined by lactate dehydrogenase(LDH) assay. The result showed that BB with the concentration of 12.5-100 μg·mL~(-1) had no toxicity to RAW264.7 and BV2 cells, and had no significant effect on the activity of cell model with low inflammation. The 50 μg·mL~(-1) BB was selected for further experiment, and the results indicated that BB inhibited LPS-induced secretion of inflammatory cytokines. The experiment on CD4~+ T cells showed that the conditioned medium for LPS-induced inflammation in BMDMs promoted the activation and differentiation of CD4~+ T cells, while the conditioned medium of the experimental group with BB intervention reduced the activation and differentiation of CD4~+ T cells. In addition, BB also enhanced the release of neurotrophic factors from BMDMs and primary microglia. The conditioned medium after BB intervention can significantly reduce the death of PC12 neurons, inhibit neuronal damage, and protect neurons. To sum up, BB plays a neuroprotective role by inhibiting macrophage and microglia-mediated inflammatory response and promoting neurotrophic factors.
Female ; Rats ; Mice ; Animals ; Bilobalides/pharmacology* ; Neuroprotection ; Lipopolysaccharides/toxicity* ; Culture Media, Conditioned/pharmacology* ; Mice, Inbred C57BL ; Macrophages/metabolism* ; Microglia ; Cytokines/metabolism* ; Nerve Growth Factors/pharmacology* ; Inflammation/metabolism*

Objective:To confirm the protective effect of Xiangsha Yuyang decoction on acetic acid-induced gastric ulcer model rats and explore its mechanism， so as to provide experimental basis for clinical drug use. Method:The 60 SPF Wistar rats were randomly divided into 6 groups： group， model group， high， middle and low dose groups of Xiangsha Yuyang decoction and omeprazole control group. The rat model of gastric ulcer was induced by acetic acid. The rats in the high， middle and low dose groups of Xiangsha Yuyang decoction were intragastrically administered at the dose of 28，14，7 g·kg^-1， and with omeprazole at the dose of 4.17 g·kg^-1in normal saline， respectively. The rats in the blank group and model group were intragastrically infused with the same volume of normal saline once a day. After 14 days of continuous treatment， the rats were killed， the blood was collected， the area and inhibition rate of gastric ulcer were measured and calculated， the histopathological sections of gastric mucosa were made and the state of gastric mucosal injury was observed， and the changes of gastric mucosal repair factor， gastric tissue related protein， oxidative stress factor and inflammatory factor in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Detected the expression of p62 Kelch-like epichlorohydrin-related protein 1 （Keap1）， nuclear transcription factor E2 related factor 2 （Nrf2） and heme oxygenase-1 （HO-1） signal pathway-related proteins in gastric mucosa by Western blot. Result:Compared with control group， the gastric mucosa of the model group showed obvious pathological changes and a large number of leukocytes infiltrated. In model group， the ulcer area was significantly increased（P<0.01）， the contents of mucin mucoprotein 5AC （MUC5AC）， epidermal growth factor （EGF）， superoxide dismutase （SOD） and increased prostaglandin E₂ （PGE₂） were significantly decreased（P<0.01）， the gastrin （GAS）， 8-hydroxydeoxyguanosine （8-OHdG）， malondialdehyde （MDA）， tumor necrosis factor-α （TNF-α）， cyclooxygenase-2 （COX-2） were significantly increased. The expression of HO-1 and Nrf2 protein decreased significantly（P<0.01）， the content of Keap1 increased significantly （P<0.05）， and the expression of p62 protein decreased. Compared with model group， the hierarchical structure of cells in Xiangsha Yuyang decoction high dose group and omeprazole group were clearer and regular， middle and low dose groups could also repair gastric mucosa to a certain extent. The high and middle dose groups of Xiangsha Yuyang decoction could significantly reduce the gastric ulcer area of acetic acid-induced gastric ulcer rat model （P<0.01） and increase the ulcer inhibition rate. It can effectively promote the expression of MUC5AC and EGF in gastric mucosa， decrease the level of GAS（P<0.05，P<0.01）， decrease the level of 8-OHdG and MDA， increase the activity of SOD（P<0.01）， decrease the expression level of TNF-α and COX-2， increase the content of PGE₂， and significantly increase the amount of Nrf2 and HO-1 protein in gastric mucosa（P<0.01）. The high dose group of Xiangsha Yuyang decoction could decrease the protein expression of Keap1（P<0.05） and increase the expression of p62 protein. Conclusion:Xiangsha Yuyang decoction is effective in the treatment of acetic acid-induced gastric ulcer model rats， which can effectively reduce the ulcer area， increase the ulcer inhibition rate and protect the ulcer tissue. Its mechanism may be related to activating p62/Keap1/Nrf2 signal pathway and regulating the expression of related genes so as to improve inflammatory response and regulate oxidative stress response.

Objective: Moxifloxacin (MFX) shows good activity against and can be a possible antibiotic therapy to treat infection; however, other studies have shown a lower or no activity. We aimed to evaluate MFX activity against using zebrafish (ZF) model . Methods: A formulation of labeled with CM-Dil was micro-injected into ZF. Survival curves were determined by recording dead ZF every day. ZF were lysed, and colony-forming units (CFUs) were enumerated. Bacteria dissemination and fluorescence intensity in ZF were analyzed. Inhibition rates of MFX and azithromycin (AZM, positive control) were determined and compared. Results: Significantly increased survival rate was observed with different AZM concentrations. However, increasing MFX concentration did not result in a significant decrease in ZF survival curve. No significant differences in bacterial burdens by CFU loads were observed between AZM and MFX groups at various concentrations. Bacterial fluorescence intensity in ZF was significantly correlated with AZM concentration. However, with increasing MFX concentration, fluorescence intensity decreased slightly when observed under fluorescence microscope. Transferring rates at various concentrations were comparable between the MFX and AZM groups, with no significant difference. Conclusion MFX showed limited efficacy against using ZF model. Its activity needs to be confirmed.
Animals ; Anti-Bacterial Agents ; pharmacology ; Disease Models, Animal ; Moxifloxacin ; pharmacology ; Mycobacterium Infections, Nontuberculous ; drug therapy ; Mycobacterium abscessus ; drug effects ; Zebrafish

Objective: Patients with opioid use disorder (OUD) have impaired attention, inhibition control, and memory function. The aldehyde dehydrogenase 2 (ALDH2 ) gene has been associated with OUD and ALDH2 gene polymorphisms may affect aldehyde metabolism and cognitive function in other substance use disorder. Therefore, we aimed to investigate whether ALDH2 genotypes have significant effects on neuropsychological functions in OUD patients undergoing methadone maintenance therapy (MMT). Methods: OUD patients undergoing MMT were investigated and followed-up for 12 weeks. ALDH2 gene polymorphisms were genotyped. Connors’ Continuous Performance Test (CPT) and the Wechsler Memory Scale-Revised (WMS-R) were administered at baseline and after 12 weeks of MMT. Multivariate linear regressions and generalized estimating equations (GEEs) were used to examine the correlation between the ALDH2 genotypes and performance on the CPTs and WMS-R. Results: We enrolled 86 patients at baseline; 61 patients completed the end-of-study assessments. The GEE analysis showed that, after the 12 weeks of MMT, OUD patients with the ALDH2 *1/*2＋*2/*2 (ALDH2 inactive) genotypes had significantly higher commission error T-scores (p = 0.03), significantly lower hit reaction time T-scores (p = 0.04), and significantly lower WMS-R visual memory index scores (p = 0.03) than did patients with the ALDH2 1 */*1 (ALDH2 active) genotype. Conclusion OUD patients with the ALDH2 inactive genotypes performed worse in cognitive domains of attention, impulse control, and memory than did those with the ALDH2 active genotype. We conclude that the ALDH2 gene is important in OUD and is associated with neuropsychological performance after MMT.

Objective To investigate the relationship between the number of colonies and the luminous intensity of two kinds of platelets contamination bacteria,Escherichia coli(ECO)and Pseudomonas aeruginosa(PAE)in single platelets, plasma and phosphate buffered salines(PBS), and to explore their growth and proliferation in different biological fluid media.Methods The two luminescent bacteria were constructed before the correlations between the number of colonies and the luminous intensity in different media in vitro were observed with a small animal bioluminescence imaging instrument, and correlation analysis was carried out.By recording the luminous intensity at serial time points in different media, a growth curve was drown to reflect the proliferation of bacteria.Results There was a significant positive correlation between the number of colonies and luminous intensity of ECO and PAE in single platelets,plasma and PBS.The larger the number of colonies was,the stronger the luminous intensity was.The results indicated that single platelets significantly inhibited the proliferation of ECO and PAE,and that plasma also had some inhibitory effect,but not so strong.Conclusion For the two luminous bacteria,luminous intensity can represent the growth and proliferation of bacteria.Single platelets have obvious inhibitory effect on the growth of ECO and PAE, and the plasma also has some inhibitory effect, but the effect is not so strong as that of single platelets.

OBJECTIVETo investigate Salmeterol/Fluticasone Propionate and Totropiumi treatment of Sillicosis merger Asthma.

METHODS30 patients with Sillicosis merger Asthma were randomly divided into group Salmeterol/Fluticasone Propionate( Single group) ( n=14) and group Salmeterol/Fluticasone Propionate and Totropiumi (Joint group) ( n= 16), patient in single group were only given Salmeterol/Fluticasone Propionate (50 f.Lg Bid) inhaling,and those in Joint group were given Salmeterol/Fluticasone Propionate (50 f.Lg Bid) and Totropiumi ( 18 f.Lg Qd) inhaling. The treatment was last for 6 months.Before the treatment,evaluation of the two groups of Sillicosis installment,determination their foungation lung function and ACT score .. After the cause of treatment, lung function FEV10/FVC(% ), FEV10 pred%, FEV10(ml), ACT score, the incidence of side effects of two groups were compared and analyzed.

RESULTThe two groups before the treatment of lung fuction and ACT score had no statistically significant difference. The two groups after treatment of lung fuction FEV10/FVC (% ),FEV10 pred%, ACT score obviously higher than before treatment (P<0.05), Joint group in FEV1/FVC(% ), ACT score significantly higher than in Single group (?<0.05), Joint group acute attack times(0.98±0.79)/time lower than Single group (2.10 ± 0.81 )/time (t=3.86,P<0.05). There were no significant side effect in two groups.

CONCLUSIONSalmeterol/Fluticasone Propionate or the combination of Salmeterol/Fluticasone Propionate and Totropiumi can improve lung function and clinical symptoms of patients with Sillicosis merger Asthma. It is also better that the combination of Salmeterol/Fluticasone Propionate and Totropiumi obviously improve clinical symptoms of patients and reduice acute attack times.

Administration, Inhalation ; Adult ; Albuterol ; analogs & derivatives ; therapeutic use ; Androstadienes ; therapeutic use ; Asthma ; complications ; drug therapy ; Drug Combinations ; Female ; Fluticasone-Salmeterol Drug Combination ; Humans ; Male ; Middle Aged ; Silicosis ; complications ; drug therapy ; Treatment Outcome

Background There is multi-dimensional order of spatial stereopsis perception in human,however,current stereopsis examination is zero-order of position disparity.A multi-dimensional space perception model is very important for the detection of stereoacuity.Objective This study was to screen the deficit of zero-order,first-order,second-order multi-dimensional spatial stereopsis perception in amblyopia and strabismus children and to explore the association of zero-order,first-order,second-order spatial perception deficit.Methods Multidimensional spacial perception was examined in 79 children aged 4-14 years in Beijing Children' s Hospital.Nineteen normal children,19 children with ametropia amblyopia,12 children with anisometropic amblyopia,18 children with strabismus and 11 children with strabismus combined amblyopia were included this study.The random-dot and line spatial stereopsis perception in zero-order,first-order and second-order were examined with a new system of multidimensional space perception screening.Written informed consent was obtained from each subject or custodian before any ocular examination associated with this study.Results Absence of zero-order,first-order,second-order random-dot channel was found in 24 children (24/79,30.4％),18 children (18/79,22.8％) and 24 children (24/79,30.4％) respectively,with an average percentage of 27.9％.Absence of zero-order,first-order,second-order line channel was examined in 37 children (37/79,46.8％),37 children (37/79,46.8％),32 children (32/79,40.5％),with an average percentage of 44.7％.In the children with a deficiency of the zero-order spatial perception,the children who still remained the first-order or/and second-order spatial perception of random-dot accounted for 41.6％ and that of lines accounted for 43.2％.In children without deficiency of zero-order random-dot or lines spatial space perception,deficiency of first-order and/or second-order spatial perception was in 37.5％ children.Various order spatial perception deficiency was seen in children suffering from amblyopia or strabismus compared with normal group(P ＜ 0.05).Conclusions There exists spatial perception deficiency in children with amblyopia or strabismus.The patients with zero-order spatial perception absence partially remain a first-order or/and second-order spatial perception;while the patients with normal zero-order spatial perception might have first-order or second-order spatial perception deficiency.The multi-dimensional space perception model has a directive role for the training of visual information process and the treatment of spatial perceptual learning in children with amblyopia or strabismus.

Through our pre-investigation and literature analysis, it was found that rhubarb could be categorized into two types, chrysophanol-type and rhein-type, based on the proportion of the two constituents in the total content of anthraquinones after acid hydrolysis. In this paper, the antimicrobial activities of chrysophanol-type and rhein-type rhubarbs against Staphylococcus aureus were compared with microcalorimetric analysis, in order to illustrate the bioactive differentiability between the two chemotypes. For the aim to display the distinction of chrysophanol and rhein percentage in total anthraquinones, the sampling volume was regulated to make the total anthraquinones equivalent, thus, the antimicrobial difference was only attributed to the difference of chemotypes. The results indicated that the antimicrobial difference between the two chemotypes was confirmable labeled at the biothermokinetic parameters of S. aureus growth affected by the rhubarb samples. The growth rate constant (k1) of the first exponential phase for the growth of S. aureus affected by the rhein-type rhubarb was significantly lower than that of chrysophanol-type (P<0.01), which suggested stronger antimicrobial activity of rhein-type rhubarb than that of chrysophanol-type. However, the antimicrobial activities of rhein-type rhubarbs were not positively correlated to the contents of rhein. It suggested that the antimicrobial activity of rhubarb might be related to some unknown components which were of same accumulating pattern of rhein. The findings in present study provided some experimental evidence on categorizing rhubarb into two chemotypes through the difference of antimicrobial activity on S. aureus by microcalorimetric analysis and, further, offered references to revision of the commercial specification of rhubarb from chemical view.
Anthraquinones ; isolation & purification ; pharmacology ; Anti-Infective Agents ; isolation & purification ; pharmacology ; Calorimetry ; Plants, Medicinal ; chemistry ; Rheum ; chemistry ; Staphylococcus aureus ; drug effects ; growth & development

OBJECTIVETo investigate the effects of Tangzhiping Granule (TZPG) on blood lipids and free fatty acids (FFA) in rats with insulin resistant diabetes (IRD).

METHODSA blank control group consisted of randomly selected normal rats was set up. The remaining rats were established to IRD model by high-fat high-sugar diet feeding and streptozotocin injection. Then the 32 successfully modeled rats were randomized into the model group (treated by saline), the Tangmaikang group (treated with Tangmaikang Granule 1.35 g/kg), and the two TZPG groups treated with high dose (2.70 g/kg) and low dose TZPG (1.35 g/kg) respectively through intragastric infusion for 4 weeks. The body weight (BW), fasting blood glucose (FBG), insulin (INS), blood lipids including triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and FFA were detected, and the insulin sensitivity index (ISI) calculated.

RESULTSCompared with the blank control group, BW, FBG and INS increased while ISI decreased in the model group (P < 0.05 or P < 0.01). All the above-mentioned abnormal indices were improved in the three treated groups (Tangmaikang, high and low dose TZPG group), but the improvements in the high dose TZPG group were more significant than those in the other two groups (P < 0.05 or P < 0.01). Similar outcomes were also seen in blood lipids detection, in which TG, TC, LDL-C and FFA were higher and HDL-C were lower in model rats than those in blank controls, they were improved in the three treated groups (P < 0.05), and the best improvements were seen in the high dose TZPG group (P < 0.05).

CONCLUSIONTZPG could reduce levels of BW, FBG, INS, TC, TG, LDL-C and FFA, and increase levels of ISI and HDL-C in rat model of insulin resistant type 2 diabetes, so as to improve the insulin resistance in them.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Acids, Nonesterified ; blood ; Female ; Insulin Resistance ; Lipids ; blood ; Male ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo analyze the clinical and laboratory features of patients with congenital dyserythropoietic anemia type I (CDA-I), and improve the clinical diagnostic accuracy.

METHODSThe clinical and hematological features of 5 patients diagnosed as CDA-I in our hospital between July 2002 and July 2007 were analyzed retrospectively, and the related literatures was reviewed.

RESULTSFive CDA-I patients, 1 male and 4 females, all had a long history of varied degree of chronic anemia. One patient had congenital malformations, 3 jaundice and 4 hepatosplenomegaly. Bone marrow specimens invariably showed hypercellularity due to erythroid hyperplasia with megaloblastic changes, irregularly shaped nuclear, and chromatin bridges in 0.2% to 0.6% of all erythroblasts. All the 5 patients' bone marrow erythroblasts showed spongy heterochromatin appearances (swiss-cheese) with electron microscopy examination. There was no morphologic abnormality in the granulocytes and megakaryocytes. Serum ferritin levels were increased in 3/4 patients. One patient had been misdiagnosed as hereditary spherocytosis and performed splenectomy in the local hospital with no improvement in Hb level.

CONCLUSIONSCDA-I is a rare congenital anemia characterized by ineffective erythropoiesis, jaundice, hepatosplenomegaly and iron overload, and may be misdiagnosed. Keeping these manifestations in mind should avoid misdiagnosis.

Adolescent ; Adult ; Anemia, Dyserythropoietic, Congenital ; blood ; diagnosis ; Female ; Humans ; Male ; Retrospective Studies ; Young Adult