Objective:To study the function and mechanism of miR-30a in rat cerebral ischemia-reperfusion (I/R) injury.Methods:The middle cerebral artery occlusion (MCAO) model was established by intravascular suture method, and the expression of miR-30a in brain tissue was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After intracerebroventricular injection of miR-30a lentivirus, the infarct area was detected by 2, 3, 5-triphenyltetrazole chloride (TTC) staining, the neurological deficit was detected by Bederson method, and the concentration of neurotrophin-3 (3-NT) and nitric oxide (NO) in brain tissue was detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of kelch like ECH associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf-2) and hemeoxygenase-1 (HO-1) in brain tissue were detected by Western blot. Double luciferase reporter assay was used to detect the targeting relationship between miR-30a and Keap1.Results:Compared with sham operation group, the expression of miR-30a was down-regulated in a time-dependent manner after I/R. The overexpression of miR-30a can reduce the area of cerebral infarction tissue at the pathological level, the degree of neurological impairment at the functional level, the 3-NT, NO and Keap1 at the molecular level, and enhance the expression of Nrf2 and HO-1. The dual luciferase reporter assay also showed that miR-30a could bind to Keap1 mRNA.Conclusions:The expression of miR-30a was down-regulated in MCAO rat brain tissue, and miR-30a could attenuate cerebral I/R injury in rats by targeting Keap1.

Adult ; Bacteremia ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Nontuberculous Mycobacteria ; Postoperative Complications

Humans ; Intestinal Obstruction/complications* ; Ileus/etiology* ; Abdominal Injuries/complications*

Objective To observe the effect of dexamethasone(Dex)on the proliferation and os- teogenic differentiation of human marrow stromal cells(MSCs)in vitro.Methods The primary human MSCs were isolated and cultured by Ficoll seperation culture in vitro.In subcultures,human MSCs were respectively treated with dexamethasone 10~(-9),10~(-8) and 10~(-7) mol/L.The proliferation of human MSCs was measured using MTF method;cytoplasmic alkaline phosphatase(ALP)activity was measured;the osteogenic marker osteopontin (OPN)mRNA were examined by reverse transcriptase polymerase chain reaction(RT-PCR).Results The op- tical density values in cultures treated with dexamethasone 10~(-8) and 10~(-7) mol/L for 8 days were significantly lower than those in the controls(P＜0.05).Treatment of cells with Dex for 12 days led to a significant increase in cytoplasmic ALP activity(P＜0.05)in a dose-dependent manner.Dex induced OPN mRNA.Conclusion Dex inhibits the proliferation of human MSCs and dexamethasone 10~(-7) mol/L leads to a strong decrease in cell number.Dex induces human MSCs differentiate to osteoblastic cells.

Objective To explore the role of clinical pharmacists in rational drug use through the pharmacy care of an elderly pneumonia patient with Chlamydia psittaci infection and drug-induced liver injury. Methods The clinical pharmacists participated in the treatment of one patient with Chlamydia psittaci pneumonia and drug-induced liver injury. Based on the results of second-generation gene sequencing, the characteristics of the pathogen were learned by literature search. The clinical pharmacists monitored the patient’s liver and kidney function, provided a new medication treatment plan to Doctors, and performed patient education during the treatment. Results The initial empirical anti-infective treatment with teicoplanin and imipenem-cilastatin was not effective. After the diagnosis of Chlamydia psittaci and Candida albicans infection, the combination of doxycycline with azithromycin and fluconazole was administered. Drug-induced liver injury was found with this treatment. The clinical pharmacist proposed to switch to doxycycline and clarithromycin with co-administration of magnesium isoglycyrrhizinate and polyene phosphatidylcholine to protect the liver. With this new regime, patient's liver function was improved and the infection was under control. Conclusion Individualized pharmaceutical cares provided by clinical pharmacists helped the safe, rational and effective use of medications.

Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
3-Hydroxybutyric Acid ; chemistry ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; Hydrolases ; genetics ; metabolism ; Hydroxybutyrates ; chemistry ; Recombinant Proteins ; genetics ; metabolism ; Rhodospirillaceae ; enzymology ; genetics

Objective To assess the feasibility of using PET molecular imaging to evaluate the therapeutic effects of traditional Chinese medicine FuZhiSan (FZS) on the model of aging Alzheimer's disease (AD) rats. Methods Twenty aged AD rats (Sparague-Dawley rats,male) were randomly divided into FZS treated group (n = 10) and control group (n = 10). Another 10 healthy adult rats were as blank controls. Morris water maze record system was used for cognitive function assessment. Before and after FZS treatment 18 F-fluorodeoxyglucose (FDG) and 11 C-2- [4'-(methylamino) phenyl] benzothiazol-6-ol ( PIB )PET imaging was undertaken. After post-treatment imaging procedures the brain tissues of all animals were taken for histochemical study,such as staining with HE,congo red,amyloid β (Aβ) immunofluorescence,5-bromo-2-deoxyuridine (BrdU) immunofluorescence and NeuN immunofluorescence. Paired t-test was performed with SPSS 13.0 software for the data analysis. Results The cognitive dysfunction of aging AD rats was improved after FZS treatment. The escape latency in FZS treated group was significantly shorter than that of control group ((32.5 ±10.8) s vs (102.6±8.8) s,t =15.7987,P=0. 0001). Diffuse neuronal loss and Aβ deposition were detected in the hippocampus and cortex in the aged AD rats. The imaging data showed that brain glucose metabolism was amended in FZS treated group while the abatement of amyloid deposition was not significant. Immunofluorescence results indicated that the neuronal proliferation was more remarkable in FZS treated group. Conclusions It may be feasible to use PET imaging as a method to evaluate the therapeutic effect in AD rats. FZS may ameliorate memory dysfunction of aged AD rats. Its mechanism may be partly contributed to the enhancement of the neuronal proliferation and survival.

Objective:To compare the two-dimensional electrophoresis(2-DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3-10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS-PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2-DE map of tissues.

OBJECTIVETo study the effect of hyperbaric oxygen (HBO) administered at different pressures and different exposure time on the differentiation of neural stem cells (NSCs) in vitro.

METHODSThe cerebral cortices from newborn rats (0-3 days old) were sterilely collected, digested, and centrifuged. After removal of the supernatant, the cells were re-suspended with DMEM/F12 medium containing B27, bFGF and EGF. The NSCs of 2-3 passages were randomly divided into seven groups: a control (untreated) and 6 HBO treatment groups that NSCs were subjected to HBO treatment of different pressures (1, 2 or 3 ATA) and different exposure time (30 or 60 minutes). The differentiated NSCs were examined by neuron-specific enolase (NSE) immunocytochemistry 24 hrs later. Percentage of NSE positive cells differentiated from NSCs was assessed by fluorescent microscopy.

RESULTSThe percentage of NSE positive cells differentiated from NSCs was the highest in the HBO 2ATA-60 min group (9.17+/-0.50%) (P<0.01), followed by the HBO 3ATA-60 min (7.89+/-0.62%), HBO 2ATA-30 min (6.72+/-0.76%), HBO 3ATA-30 min (6.08+/-0.57%), HBO 1ATA-60 min (5.45+/-0.52%), HBO 1ATA 30 min (3.85+/-0.44%) and control groups (3.72+/-0.88%). In addition to the HBO 1ATA-30 min group, the other HBO treatment groups had increased significantly percentage of NSE positive cells compared with the control group (P<0.01). Under the same pressure, the 60 min treatment groups had increased significantly percentage of NSE positive cells compared with the 30 min treatment groups (P<0.01).

CONCLUSIONSHBO treatment (2 ATA, 60 minutes) produces a best effect in the differentiation of NSCs into neurons.

Animals ; Animals, Newborn ; Cell Differentiation ; Hyperbaric Oxygenation ; Neurons ; cytology ; Pressure ; Rats ; Stem Cells ; cytology ; Time Factors

Objective To assess the role of 68Ga-N,N′-bis(2-hydroxy-5-(carboxyethyl)benzyl) ethylenediamine-N,N′-diacetic acid(HBED-CC)-(Ahx)Lys-CO-Glu(PSMA-11) PET/CT on the detection of metastatic lesions from castration-resistant prostate cancer (CRPC).Methods Sixteen patients with CRPC who underwent 68Ga-PSMA-11 PET/CT between January 2015 and November 2015 were recruited in this study.Mean age of patients was (72±9) years.The PSA levels were 4-12 356 μg/L, Gleason score was 7-10.PET/CT was performed at 1 h postinjection of 68Ga-PSMA-11.Patient-based analysis and lesion-based analysis were performed.ROI analysis was used to calculate the tumor uptake (SUVmax).Final diagnosis was based on histopathology and results of other imaging examinations(99Tcm-MDP imaging, MRI).χ2 test was used to compare the diagnostic efficiencies of PET and CT.Results No adverse effects were observed in patients.68Ga-PSMA-11 PET/CT showed moderate physiologic uptake in salivary glands and proximal small intestine, with predominant tracer clearance by the kidneys.All patients were positive on 68Ga-PSMA-11 PET/CT.Bone metastasis was found in 16 patients, liver metastasis in 2 patients (5 lesions), and lymph node metastasis in 4 patients (26 lesions).The SUVmax of liver, lymph node and bone metastases were 15.06±2.77, 7.54±5.20, 19.01±16.96, respectively.The diagnostic sensitivity, specificity and accuracy on bone metastasis with 68Ga-PSMA-11 PET and CT were 96.30%(52/54) vs 61.11%(33/54), 3/3 vs 1/3, 96.49%(55/57) vs 59.65%(34/57).The sensitivities and accuracies of the two modalities were significantly different(χ2=19.943, 22.593, both P<0.01).Conclusions 68Ga-PSMA-11 PET/CT could precisely detect both primary and metastatic lesions of CRPC, suggesting that it is of great value for the clinical management and treatment.