OBJECTIVETo analyze the electrical activity property changes in nucleus accumbens (NAc) of heroin-induced conditioned place preference (CPP) rats during different stages of heroin dependence and to explore NAc's roles in the formation of drug dependence.

METHODSRecording electrodes were bilaterally embedded into the NAcs of rats with the aid of stereotaxic apparatus, followed by establishment of heroin-dependent rat model. The NAc electrical activity during 3 different stages of heroin dependence, including heroin pre-exposure, immediate post-exposure and heroin withdrawal, were respectively recorded using EEG wireless telemetry techniques. The frequency distribution (ranging from 0.5 to 30 Hz) and the amplitude of NAc electrical activity were analyzed and measured.

RESULTSHeroin-dependent rat models were successfully established and their withdrawal symptoms were evident. All rats showed a conditioned place preference (CPP) for the white box after 5-10 days of heroin-exposure, and displayed a maximum withdrawal symptoms on 2d after heroin- withdrawal. During all statges of heroin-dependence, the NAc electrical activity contained the highest proportion of delta rhythm and the lowest proportion of alpha2 rhythm. The discharge frequence band was similar across different stages. There was a significantly increased ratio of low-frequency discharges (delta rhythm) and decreased ratio of high-frequency discharges (beta rhythm) in NAc of rats during the immediate post- heroin exposure stage when compared with that during pre-exposure and heroin withdrawal stages. During the withdrawal stage, the ratio of at rhythm was significantly lower than during pre- and post-heroin exposure stages (P < 0.01). Further, the mean discharge amplitude in NAcs during immediate post-exposure and withdrawal stages was significantly increased relative to pre-exposure stage. However, the mean discharge amplitude during heroin withdrawal stage was significantly lower than during immediate post-exposure stage.

CONCLUSIONThe electrical activity properties in rat NAcs showed a significant change during different stages of heroin-dependence, which suggested that neuronal activities in NAcs might contribute to the modulation of drug-dependence.

Animals ; Conditioning, Operant ; Heroin ; pharmacology ; Heroin Dependence ; physiopathology ; Male ; Nucleus Accumbens ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Telemetry

Aim To observe the influences of dexmen-detomidine on the spontaneous contraction of duodenal smooth muscle of rabbits in vitro and explore the mech-anisms.Methods The rabbits ( male or female ) were stunned and the duodenums were isolated .The sam-ples of duodenal segments were connected with tension transducer , which were then put into oxygen saturation Krebs-Henseleit ( K-H) solution .The influences of dex-mendetomidine on amplitude ( AM ) and frequency ( FR ) of duodenal smooth muscle were recorded by BL-420 F biological signal processing system .The cu-mulative dosing method was used to observe the differ-ent concentrations of dexmedetomidine on duodenal smooth muscle spontaneous contraction .Glibenclamide ( Gli) was added to K-H solution before dexmendeto-midine.In the calcium-free K-H solution, calcium chloride and rynodine were added before dexmendeto-midine.The mechanisms of dexmendetomidine were studied .Results ① Dexmendetomidine reduced the amplitude of spontaneous contraction of duodenal smooth muscle in rabbits in a dose-dependent manner ( P<0.05 or P<0.01 ) , while the frequency was not obviously influenced ( P >0.05 ) .② Gli ( P <0.05 ) partly abolished the inhibitory effects of dexmendetomi-dine on duodenal smooth muscle .③ Dexmendetomi-dine inhibited the contraction of duodenum smooth muscle induced by calcium chloride ( P <0.05 ) and rynodine ( P<0.05 ) application into calcium-free K-H solution.Conclusion Dexmendetomidine inhibits the spontaneous contraction of duodenal smooth muscle of rabbits in vitro.The mechanisms may be related to ac-tivating ATP sensitive potassium channels , inhibition of the extracellular calcium influx via cell membrane and intracellular calcium release via sarcoplasmic reticulum in duodenal smooth muscle .

OBJECTIVETo observe the effects of auricular acupuncture on the learning and memory abilities of model rats with Alzheimer's disease (AD), and investigate its mechanism.

METHODSThirty SD rats were randomly divided into a control group, a model group and an auricular acupuncture group, 10 rats in each group. The model rats with AD were established by multiple injections with Okadaic Acid into the CA1 region of hippocampus. In the control group, the same quantity injection with Dimethyl Sulfoxide (DMSO) was applied on experimental rats. The auricular acupoints of "Nao" (brain) and "Shen" (kidney) were used for treating in the auricular acupuncture group, in contrast, the auricular region were not treated in the model and the control groups. The learning and memory capabilities of the rats were assessed with Morris Water Maze behavioral test, and the expressions of choline acetyltransferase (ChAT) and glial fibrillary acidic protein (GFAP) were examined by immunohistochemistry.

RESULTSComparing with the model group, the treated AD rats with auricular acupuncture was showed that the average escape latency was obviously shortened in the place navigation test (P<0.01), the movement time in plateform quadrant was obviously prolonged in the spatial probe test (P<0.05), and the number of traversing platform obviously increased (P<0.01) after the platform was taken away. The expression of ChAT increased in the hippocampus and cortex (P<0.01, P<0.05), but the expression of GFAP obviously decreased in the CA1 region of hippocampus (P<0.01).

CONCLUSIONAuricular acupuncture can improve the learning and memory capability of the model rats with AD. Its mechanism might be related with decreasing cholinergic neuron damage and reducing the abnormal activation and hyperplasia of astrocyte.

Acupuncture, Ear ; Alzheimer Disease ; genetics ; metabolism ; psychology ; therapy ; Animals ; Choline O-Acetyltransferase ; genetics ; metabolism ; Disease Models, Animal ; Gene Expression ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Humans ; Male ; Memory ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship between survivin mRNA and protein expression and nasopharyngeal carcinoma (NPC).

METHODSSurvivin mRNA and protein were detected by in situ hybridization and immunohistochemical S-P staining respectively.

RESULTSAmong 64 cases of NPC, 42 cases (65.6%) were positive for survivin mRNA expression, 30 cases (46.9%) had high expression level. In 46 cases (71.9%) of NPC were positive for survivin protein expression, and 38 cases (59.4%) had high expression level. In 22 cases of NPC with detailed clinical information, the positive expression rates of survivin mRNA and protein in stages III+ IV of NPC were 66.7% and 61.1% respectively, which were higher than those in stages I+ II of NPC (50.0% and 50.0% respectively). There was no significant difference between survivin mRNA and protein expression regarding age or gender of NPC patients (all P> 0.05). The positive expression rates of survivin mRNA and protein in chronic nasopharyngitis group were 33.3% and 23.3% respectively, which were lower than those in NPC group (chi (2)= 12.04, P< 0.01 and chi (2)= 19.57, P< 0.01, respectively). In 64 cases of NPC, 36 cases were positive for both survivin mRNA and protein, and the expression of survivin mRNA and protein showed positive correlation (phi = 0.43).

CONCLUSIONThe expression of survivin gene may play some roles in the pathogenesis of NPC. Detection of survivin mRNA and protein will be helpful for diagnosis, clinical staging and prognosis of NPC.

Adult ; Age Factors ; Aged ; Carcinoma ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; Sex Factors ; Young Adult

AIM:To investigate the inhibitory effect of apolipoprotein A-I mimetic peptide D-4F on the scaven-ger receptor A1 ( SR-A1 ) in macrophage-derived foam cells induced by oxidized low-density lipoprotein ( ox-LDL ) . METHODS:RAW264.7 cells were pretreated with different concentrations (12.5, 25 and 50 mg/L) of D-4F or 50 mg/L inactive control peptide scrambled D-4F (sD-4F) for 1 h or endoplasmic reticulum stress (ERS) inhibitor 4-phenylbutyr-ic acid (5 mmol/L) for 30 min, followed by the treatment with 100 mg/L ox-LDL for 12 h.In addition, the cells were pre-treated with 50 mg/L D-4F or sD-4F for 1 h, and then stimulated with 2 mg/L tunicamycin (TM;an ERS inducer), for 4 h.The viability of the cells was measured by MTT assay, and the content of intracellular total cholesterol ( TC) was meas-ured by a tissue/cell TC assay.The protein and mRNA levels of SR-A1 and glucose-regulated protein 78 (GRP78) were analyzed by Western blotting and quantitative real-time PCR, respectively.The fluorescence intensity of DiI-ox-LDL in the cells was detected by a multifunctional microplate reader.RESULTS:D-4F significantly reduced ox-LDL-induced macro-phage injury and intracellular cholesterol accumulation, and attenuated the ox-LDL-induced expression of SRA1 and GRP78 in a dose-dependent manner.Additionally, D-4F significantly inhibited the TM-induced protein expression of SR-A1 and GRP78, and attenuated the uptake of ox-LDL by macrophages.CONCLUSION: D-4F reduces ox-LDL-induced macro-phage cholesterol accumulation and injury by inhibiting SR-A1 expression.The mechanism may be related to the inhibition of ERS signaling pathway mediated by GRP78.

Objective To investigate drug resistance genes and epidemic characteristics of β-lactamase carried by carbapenem-resistant Acinetobacter baumannii (CRAB) in the respiratory intensive care unit(RICU) in a hospital.Methods Clinically isolated CRAB from RICU patients in October-December 2015 were collected.Five drug resistance genes (KPC-2,IMP,VIM,NDM-1,OXA-23) were specifically amplified by polymerase chain reaction (PCR),amplified products were performed agarose gel electrophoresis and sequencing analysis,the homology was analyzed with pulsed-field gel electrophoresis (PFGE).Results A total of 22 CRAB strains were isolated in October-December 2015,19 (86.36％) of which were isolated from sputum.The resistance rate of 22 CRAB strains to compound sulfamethoxazole was 59.09 ％,resistance rate to minocycline was 9.09 ％,all were sensitive to polymyxin B,resistance rates to other antimicrobial agents were more than 80％.Three kinds of resistance genes KPC-2,IMP and NDM-1 were not found by PCR amplification,positive rates of VIM and OXA-23 were both 100％.PFGE homology analysis revealed that 22 strains were divided into 13 different types,each type contained 1-5 strains,9 types(69.23％) contained only 1 strain respectively,the other 4 types (30.77％) contained 2-5 strains.A5,A7,and A8;A9,A11,A14,A19 and A22;A4,A10 and A12;A16 and A18 were of the same type respectively.Conclusion The main types of β-lactamase-resistant genes of CRAB in RICU are VIM and OXA-23.Homology analysis shows a small parts are of the same clone strains,which reveals epidemic of a small scale.

OBJECTIVE: To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice, we have designed a new method to produce standard D1S549 allelic ladder. METHODS: Eight different PCR amplified D1S549 allelic fragments were isolated from the gel, eluted into the distilled water and re-amplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors and transfected into competent E. coli DH5 alpha cells. RESULTS: The sequencing results confirmed that the size and the construe of the inserts were correct. The recombinant plasmids DNA with 8 inserts were then used as templates for re-amplification to generate D1S549 standard ladder, with which the genetic polymorphisms of D1S549 locus in Chinese Han population in chengdu, Hui population in Gansu and Wei population in Xinjiang were studied. CONCLUSION The results showed that the standard ladder made via this method is excellent, and D1S549 locus is robust for genetic research and forensic application.
Alleles ; China ; Cloning, Molecular ; Forensic Medicine ; Gene Frequency ; Genetics, Population ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Tandem Repeat Sequences/genetics*

Objective To determine the effects of naloxone on the withdrawal symptom in heroin-induced conditioned place preference (CPP) rats and the spontaneous firing properties in nucleus accumbens (NAc).Methods The heroin-dependent rat models were established,and the recording electrodes were implanted to NAc via stereotaxic apparatus.After 4 days of natural withdrawal,rats were injected with naloxone (0.5mg· kg-1),after which the withdrawal symptoms were evaluated.The electrical activities of NAc neurons were recorded by wireless telemetry at the following stages:before modeling,immediately after heroin administration,and heroin withdrawal.The frequency distribution at 0.5 ～ 30.0 Hz of the spontaneous firing was arranged and compared with that after naloxone administration.Results The heroin-dependent rat model was effectively established through the CPP system.Rats showed significant withdrawal symptoms with dynamic characteristics,which appeared the most significant on the first two days of withdrawal (6.6 ± 1.7) point,compared with before modeling with statistical difference (P ＜0.01).After naloxone irritability,the withdrawal symptoms reached the highest level(7.2 ± 1.3) point,compared with before modeling with statistical difference(P ＜ 0.01).In each state,δ wave occupied the largest ratio in the spontaneous discharge,while β wave held the smallest proportion.However,the distribution gradient of the different electroencephalogram(EEG) in each state demonstrated similar characters.Compared with the state immediately after heroin administration ofδ wave (45.58 ± 7.80)％ and θ wave (22.67 ± 3.37)％,the ratio of δ wave (38.58 ±5.76％),θ wave (18.75 ±3.31)％ in the state of naloxone irritation with statistical difference(P ＜0.05).Compared with the state immediately after heroin administration of the ratio of β31 wave (7.50 ±2.28)％ and β2 wave (10.17 ± 3.33) ％,the ratio of β1 wave (11.00 ± 1.35) ％ and β2 wave (16.50 ± 2.15) ％ in the state of naloxone irritation with statistical difference (P ＜ 0.01).In summary,after naloxone irritability,EEG energy state showed the largest difference with the state immediately after heroin administration,and differed less with the state before heroin administration.Conclusion Naloxone can stimulate the withdrawal symptoms in heroin-dependent rats,and significantly affect the electrophysiological properties in NAc,which implies that naloxone exerts an important role in NAc energy state in drug-dependent rats.

Objective To evaluate the chemopreventive effects of boswellic acid and curcumin on 7,12-dimethyl benzanthracene (DMBA)-induced oral carcinogenesis in the hamster cheek pouch model.Methods Male Syrian golden hamsters(6-8 weeks old,80-130 g in weight) were randomly divided into seven groups,with group A serving as the untreated negative control.The left cheek pouch of the remaining hamsters was topically treated with 0.5％ DMBA in mineral oil three times a week for 6 weeks.They were then randomized to six groups with group B serving as a positive control and receiving no further treatment.Groups C-G were treated topically with 5,10 mg/L boswellic acid,5,10 μ moL/L curcumin,or the combination of 5 mg/L boswellic acid and 5 μmnol/L curcumin three times per week for 18 weeks.The animals were injected with bromodeoxyuridine intraperitoneally at 50 mg/kg 2 h prior to killing.At the 25 th week all the hamsters were sacrificed and cheek pouch tissue was harvested.One half of the tissue was snap frozen in liquid nitrogen for analysis of arachidonic acid metabolites,and the other half was fixed in 10％ phosphate-buffered saline(PBS)-buffered formalin for histopathological examination.Results Six-weeks of DMBA followed by 18-weeks of topical application of boswellic acid and curcumin,both boswellic acid (5,10 mg/L) and curcumin (5,10 μmol/L) significantly inhibited the incidence from 93.8％ to 73.9％ (P＞0.05),numbers from 2.19 ± 0.98 to 1.13 ± 0.81 (P ＜ 0.01) and size of visible tumors.Microscopically the incidence of squamous cell carcinoma and BrdU index were also significantly suppressed by boswellic acid and curcumin.Conclusions Both boswellic acid and curcumin were effective in preventing oral carcinogenesis in DMBA-induced hamster cheek pouch model.

The purposes of the present study were to investigate the inhibitory effect of quercetin (QUE) preconditioning on endoplasmic reticulum stress (ERS) inducer tunicamycin (TM)-induced apoptosis in RAW264.7 macrophages and the underlying molecular mechanisms. RAW264.7 cells were pretreated with different concentrations (20, 40, and 80 μmol/L) of QUE for 30 min and then treated with TM (5 mg/L) for 12 h. Cell viability and apoptosis were determined using MTT assay and Annexin V-FITC apoptosis detection kit, respectively. The nuclear translocation of activating transcription factor 6 (ATF6) in cells was detected by immunofluorescence analysis and Western blot. Protein and mRNA expressions of C/EBP homologous protein (CHOP) and Bcl-2 were examined by Western blot and real-time PCR, respectively. The results showed that TM reduced cell viability and induced apoptosis in RAW264.7 macrophages. The cytotoxic effects of TM were significantly inhibited by QUE pretreatment at the concentrations of 40 and 80 μmol/L. Interestingly, we found that QUE also significantly suppressed the TM-induced translocation of ATF6, an ERS sensor, from the cytoplasm to the nucleus. In addition, exposure of RAW264.7 macrophages to TM resulted in a significant increase of the expression of CHOP, a transcription factor regulated by ATF6 under conditions of ERS, as well as a decrease of Bcl-2 at transcript and protein levels. QUE blocked these effects in a dose-dependent manner. These data indicate that QUE can protect RAW264.7 cells from TM-induced apoptosis and that the mechanism at least partially involves its ability to inhibit the ATF6-CHOP signaling pathway.
Activating Transcription Factor 6 ; metabolism ; Animals ; Apoptosis ; Cell Survival ; Endoplasmic Reticulum Stress ; Macrophages ; cytology ; drug effects ; Mice ; Quercetin ; pharmacology ; Transcription Factor CHOP ; metabolism ; Tunicamycin ; pharmacology