Objective To investigate changes of synaptic interface structural in the hippocampus Cal in high and low conditioned place preference(CPP) rats after chronic morphine treatment.Methods The male SD rats were randomly distributed to experiment group 130 cases(intraperitoneal injected morphine twice a day for ten days in an ascending dosage schedule) and control group 30 cases(injected saline of the same volume at same time).The rats in experiment group were re-classified into high preference group(HP),middle group and low preference group(LP) according to the numerical value of the CPP.The middle group was rejected.The rats in HP and LP were scarified at the time of 3h,3d and 14d after the last injection.The hippocampus Cal were removed and prepared for electron microscope specimen.The synaptie interface structure parameter were analyzed by image processing technique.Results ①No significant difference of pretest scores staying at the non-preference chambet existed among the three groups(F=0.78,P=0.47).However,the test scores of the CPP minus the time stayed at pretest natural preference in the high group was significantly higher than that of the low group(P=0.00).②At the 3h and 3d,the PSD of the high group((15.20±-3.65)nm) was significantly lower than low group((17.63±6.61)nm,P<0.01);the synaptic cleft of high group((5.77±2.08)nm) was significantly higher than low group ((4.92±1.65)nm,P<0.05).At the 14d,the PSD of the high group((16.22±4.93)nm) was significantly lower than low group((18.42±3.78)nm,P<0.01).Conclusion In hippocampal Cal area the synaptic cleft in the HP group was higher than that of LP group,the post-synaptic density in the HP group was lower than that of LP group.These changes may be the synaptic basic of the different susceptibility.

Objective This study is to investigate the changes in the NFATc 4/3 signaling pathway in rat hippocampus after whole brain radiation. Methods A total of 120 one?month?old male Sprague?Dawley rats were randomly divided into four groups to receive whole brain radiation using 4?MeV electron beams with doses of 0( control) ,2,10,and 20 Gy,respectively,in a single fraction. At 6 hours,12 hours,1 day,3 days,1 week,and 2 weeks after radiation,Western blot and real?time PCR were used to evaluate the changes in expression levels of CaN, NFATc 4/3, p?NFATc 4/3, and GSK?3β. Results There were no significant changes in the expression of NFATc 4/3 or p?NFATc 4/3 at 6 and 12 hours after whole brain radiation. At 1 day after radiation,compared with the control group,the expression of p?NFATc 4/3 in the radiation groups was significantly increased in a dose?dependent manner ( 2 Gy:P= 0. 014;10 Gy:P=0. 011;20 Gy:P=0. 000 );however, there was no significant difference in the expression of NFATc 4/3 between the radiation group and the control group. The expression of NFATc 4/3 was significantly decreased in the radiation groups than in the control group at day 3 ( 2 Gy:P=0. 040;10 Gy:P=0. 000;20 Gy:P=0. 000),1 week (2 Gy:P=0. 692;10 Gy:P=0. 032;20 Gy:P=0. 021),and 2 weeks (2 Gy:P=0. 001;10 Gy:P=0. 000;20 Gy:P=0. 000) after radiation,while there was no significant difference in the expression of p?NFATc 4/3 between any two groups. There were no time?or dose?dependent changes in expression of CaN or GSK?3β. Conclusions Ionization radiation has an inhibitory effect on the NFATc 4/3 signaling pathway in rat hippocampus. Combined with our previous results,this study suggests that radiation?induced cognitive dysfunction is associated with the NFATc 4/3 signaling pathway.

Objective To investigate the effects of hypoxia-inducible factor-1α (HIF-1α)expression on pathogenesis of retinopathy of prematurity (ROP) and to find new target for gene therapy.Methods After liposome-mediated small interference RNA (siRNA) transfection into rat retinal endothelial cells,the cells were cultured in medium with CoCl2-induced hypoxic condition.Expression of HIF-1α mRNA was determined by fluorenscence quantitative reverse transcription-polymerase chain reaction(RT-PCR),HIF-1α protein expression was detected by Western Blot after cocultured for 8 hours.Cell proliferation was measured with 3-(4,5)-dimethylthiazol (-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay after cocultured for 24 hours.Difference between groups was compared with independent samples t test.Results Rat retinal vascular endothelial cells were successfully transfected with siRNA.Fluorescence quantitative RT-PCR results showed that at 48 hours of transfection,the expression of HIF-1α mRNA in the interference group of siRNA1,siRNA2 and siRNA4 were 0.1620 ± 0.0147,0.2034 ± 0.0251 and 0.3049 ± 0.0165,which were 16.20％,20.34 ％ and 30.49％ of blank control group (1.0000±0.0344),and were lower than that of negative control group (0.8334±0.0242) (t=16.786,8.953 and 4.087,P＜0.05 respectively).Western Blot results showed that HIF-1α protein expression was significantly inhibited by siRNA1(0.4956 ± 0.0421 ) and siRNA2 (0.6544 ± 1.0032) comparing with blank control group (3.5105 ±0.4084) and negative control group (3.4019 ± 1.0677) (t =6.861,2.893,4.567 and 5.072,P＜0.05 respectively).As for cellular proliferation activity,(49.5±2.9) ％ and (67.4±1.2) ％ of cells growth inhibition were observed after transfection with siRNA1 and siRNA2,which were higher than those of negative control group [(15.7±1.5) ％ ] (t=2.786 and 6.904,P＜0.05).Conclusions The synthetic HIF-1α siRNA could effectively inhibit the expression of HIF-1α gene and reduce cell proliferation in rat retinal endothelial cells under hypoxic condition.RNA interference technology targeting HIF-1α might become a new strategy for gene therapy of ROP.

Objeetive To explore the differentiation of B lymphocytes and expression of B7-related protein-1 (B7RP-1)on B lymphocytes in patients with systemic lupus erythematosus(SLE).Methods Three-color immunofluorescent staining and flow cytometric assay were used to analyze the frequency of three types of B lymphocytes,I.e.,plasma cells,memory B lyphocytes and naive B lymphocytes,as well as the expression of B7RP-1 on these cells in peripheral blood from 23 patients with SLE and 16 normal human controls.Clinical data of these patients with SLE were collected.and SLE disease activi index(SLEDAI)was also evaluated.The relationship was assessed between the expression of B7RP-1 and SLEDAI.Results The frequency of plasma cells was highest in patients with active SLE.followed by patients with inactive SLE and normal human controls(P＜0.01).A significant decrease was observed in the frequency of memory B lymphocytes in patients with active SLE compared with normal controls (P＜0.01),but no significant difference was found between patients with inactive SLE and those with active SLE(P＞0.05).Regarding the frequency of naive B lymphocytes,there was no significant difierence among the three groups.Increased frequency of plasma cells was also noted in patients with lupus nephritis(LN)compared with those without LN [(6.15±3.12)%vs(3.31±1.41)%,P＜0.05 ],but no significant difierence was found with regard to the frequency of memory or naive B lymphocytes between these two groups.The expression rate Of B7RP-1 was significantly lower on total lymphocytes from patients with SLE than from normal human controls (46.51%vs 63.75%,P＜0.05),which was the case with B7RP-1 on plasma cells,memory B lyphocytes and naive B lymphocytes (all P＜0.01),whereas no significant difierence was found between patients with inactive SLE and active SLE or between patients with and without LN.In addition.no correlation was found between the expression of B7RP-1 and SLEDAI(r=0.035,P＞0.05).Conclusions In peripheral blood of patients with SLE,the frequency of plasma cells is increased,while the expression of B7RP-1 on lymphocytes is decreased,which may be relevant to the pathogenesis of SLE.

AIM: To investigate the effect of retinoic acid on proliferation of renal interstitial myofibroblasts in a rat model of the left unilateral ureteal obstruction (UUO). METHODS: UUO model was established by unilateral ligation of ureter in 36 SD rats. Rats were divided into 2 groups: Retinoic acid-treated group and control group, each with 18 rats. UUO rats were treated with either daily subcutaneous injection of 10 mg/kg body weight of all trans-retinoic acid or vehicle alone two days before the operation until being sacrificed. Groups of 6 rats were killed on day 3, 7 and 12 after ligation of the left ureter. The percentage of renal tubular lesion, interstitial fibrosis score, the number of interstitial myofibroblasts, the number of proliferating myofibroblasts and the expression of TGF?-1 mRNA were determined. RESULTS: There were significant accumulation and local proliferation of myofibroblasts in the interstitium of the UUO rats. On day 7 of the UUO model, the percentage of tubular lesions and interstitial fibrosis score were significantly lower in the retinoic acid-treated group than those in control group [(15.9?2.0)% vs (27.3?2.2)% and (0.47?0.12) vs (1.65?0.18), P

Objective To explore the empathy deficits of adolescents with different types of conduct disorder. Methods The participants included 65 adolescents ( who met the DSM-Ⅳ criteria for conduct disorder)ranging from 13 ～ 18 in age,and 195 normal adolescents ranging from 13 ～ 18 in age. All participants were assessed by revised Basic Empathy Scale. 65 patients were divided into 4 subgroups according to symptoms: destructive-nondestructive subgroups and overt-covert subgroups. Results (1) Scores of cognitive empathy were lower in patients than normal controls ( (29.86 ± 4.72) vs ( 32.09 ± 4.94), P ＜ 0. 01 ). (2) There were significant differences in the levels of cognitive empathy between patients and controls (P＜0. 01 ). Scores of cognitive empathy were lower in destructive subgroup than controls ( (29.76 ± 4.46) vs ( 32.09 ± 4.94) , P ＜ 0.01 ). (3) There were significant differences in the levels of two dimensions and total scores of empathy between patients and controls (F=3.10 ～5.36, P ＜ 0.05 ). Scores of cognitive empathy were lower in overt subgroup ( 29.22 ± 3.77 ) and covert subgroup (30.21 ± 5.17 ) than controls (32.09 ± 4.94) (P＜ 0.05, P＜0.01 );and scores of affective empathy were lower in overt subgroup than covert subgroup ( (26.13 ±5.05) vs (29.50 ±4.16), P＜0.05 ). Total scores of empathy were lower in overt subgroup (55.35 ±7.09) than covert subgroup (59.71 ±7.58) and controls (60.04 ±8.50 ) (P＜0.05). (4) Logistic regression analysis showed that cognitive empathy was a protective factor for conduct disorders( OR = 0.43 ). Conclusion The cognitive empathy level of patients is significantly lower than normal students;and boys with different types of conduct disorder possess different empathic abilities when compared with healthy controls;and cognitive empathy is a protective factor for conduct disorder.

OBJECTIVEThis study investigates the circadian variation rules of the clock gene Per2 and clock-controlled genes of vascular endothelial growth factor (VEGF), Ki67, c-Myc, and P53 in different stages of carcinogenesis in buccal mucosa carcinoma and their roles in the development of buccal mucosa carcinoma.

METHODSNinety Syrian golden hamsters were housed under. 12 h light/12 h dark cycles. Dimethylbenzanthracene (DMBA) was used to establish the carcinoma model by smearing the golden hamster buccal mucosa. Before DMBA painting and after 6 and 14 weeks, the hamsters were sacrificed at six time points within a period of 24 h (i.e., 4, 8, 12, 16, 20, and 24 h after light onset), and the normal buccal mucosa, precancerous lesions, and cancer tissues were simultaneously obtained. Hematoxylin and eosin stained sections were prepared to observe the canceration of each tissue. Real time polymerase chain reaction was used to detect the mRNA expression of Per2, VEGF, Ki67, c-Myc, and P53. Cosine analysis was employed to determine the circadian-rhythm variations of Per2, VEGF, Ki67, c-Myc, and P53 mRNA expression in terms of median, amplitude, and acrophase.

RESULTSThe expression of Per2, VEGF, P53, and c-Myc mRNA in three different stages appeared with circadian rhythms (P<0.05), whereas the Ki67 mRNA was expressed with circadian rhythm only in normal and precancerous lesion stages (P<0.05). The midline-estimating statistic of rhythms (MESORs) of Per2 and P53 mRNA were significantly down-regulated with the development of cancer (P<0.05), whereas the MESORs of VEGF, c-Myc, and Ki67 mRNA were up-regulated (P<0.05). The amplitude of P53 mRNA significantly decreased with the development of cancer (P<0.05). Moreover, compared with the normal group, the amplitudes of Per2, VEGF, Ki67, and c-Myc mRNA significantly increased in precancerous lesions and cancer tissue (P<0.05). In precancerous stage, the acrophases of Per2, VEGF, and c-Myc mRNA were earlier than that in the normal group, whereas that of Ki67 and P53 mRNA were delayed.

CONCLUSIONThe circadian-rhythm characteristics of the clock gene Per2 and clock-controlled gene expression of VEGF, Ki67, c-Myc, and P53 mRNA have changed with the occurrence and development of carcinoma.

9,10-Dimethyl-1,2-benzanthracene ; Animals ; Carcinogenesis ; Carcinoma, Squamous Cell ; metabolism ; Circadian Rhythm ; Cricetinae ; Mesocricetus ; Mouth Mucosa ; metabolism ; Mouth Neoplasms ; metabolism ; Neoplasm Staging ; Period Circadian Proteins ; genetics ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A

Objective To evaluate effects of a fusion protein LTβR-Fc, which can block the herpesvirus entry mediator ligand (LIGHT-HVEM)signaling pathway, on ovalbumin-induced dermatitis in a mouse model. Methods Thirty BALB/c mice were randomly and equally divided into 3 groups: blank control group treated with 100 μl of sodium chloride physiological solution, model group sensitized with 100 μl of sodium chloride physiological solution containing 100 μg ovalbumin, blocker group firstly blocked with 100 μl of sodium chloride physiological solution containing 100 μg LTβR-Fc followed by sensitization with 100 μl of sodium chloride physiological solution containing 100 μg ovalbumin at 24 hours after the blocking. Disease severity was evaluated by eczema area and severity index (EASI)score, and lesional size was measured on day 0, 4, 8, 12, 15, 20, 23, 27, 31 and 34 after the first sensitization. A total of three sessions of sensitization were carried out. At the end of treatment, all the mice were sacrificed after serum was obtained from their orbital cavities. Thereafter, tissue specimens were obtained from skin lesions, and single cell suspensions of the spleen were prepared. RT-PCR was performed to detect mRNA expressions of interferon γ (IFN-γ), interleukin 4 (IL-4)and IL-5 in murine lesions, ELISA to measure IFN-γ, IL-4 and IL-5 levels in culture supernatants of murine splenocytes, as well as ovalbumin-specific and total IgE and IgG1 levels in murine sera. Results LTβR-Fc significantly suppressed inflammatory response in the mouse model of dermatitis induced by ovalbumin. Compared with the model group, the blocker group showed significantly decreased lesion area and EASI score (both P < 0.05). In addition, a significant decrease was observed in the mRNA expressions of IL-4 (0.88 ± 0.25 vs. 1.81 ± 0.25, P < 0.05), IL-5 (0.75 ± 0.15 vs. 1.24 ± 0.26, P < 0.05)and IFN-γ (0.62 ± 0.09 vs. 1.11 ± 0.19, P < 0.05)in murine lesions, and in supernatant levels of IL-4 (9.58 ± 1.44 ng/L vs. 20.12 ± 5.39 ng/L, P < 0.05), IL-5 (11.37 ± 2.02 ng/L vs. 22.77 ± 4.07 ng/L, P < 0.05)and IFN-γ (16 167 ± 950.40 ng/L vs. 23 930 ± 44.20 ng/L, P < 0.05)in the blocker group compared with the model group. The serum levels of both total IgE and ovalbumin-specific IgE were significantly lower in the blocker group than in the model group(total IgE: 27 466.67 ± 2 052.64 μg/L vs. 32 277 ± 407.53 μg/L, P < 0.05; ovalbumin-specific IgE: 1 296.33 ± 32.72 μg/L vs. 2 323.33 ± 502.43 μg/L, P < 0.05), so were those of total IgG1 (0.46 ± 0.11 μg/L vs. 0.84 ± 0.11 μg/L, P < 0.05)and ovalbumin-specific IgG1 (0.62 ± 0.11 μg/L vs. 0.86 ± 0.07 μg/L, P < 0.05). Conclusion The fusion protein LTβR-Fc can alleviate symptoms of ovalbumin-induced dermatitis in the mouse model likely by suppressing the LIGHT-HVEM signaling pathway, suggesting that this signaling pathway may serve as a target for the treatment of dermatitis(such as atopic dermatitis).

Neurons in the laterodorsal tegmentum (LDTg) and pedunculopontine tegmental nucleus (PPTg) play important roles in central autonomic circuits of the kidney. In this study, we used a combination of retrograde tracers pseudorabies virus (PRV)-614 and fluorescence immunohistochemistry to characterize the neuroanatomic substrate of PPTg and LDTg innervating the kidney in the mouse. PRV-614-infected neurons were retrogradely labeled in the rostral and middle parts of LDTg, and the middle and caudal parts of PPTg after tracer injection in the kidney. PRV-614/TPH double-labeled neurons were mainly localized in the rostral of LDTg, whereas PRV-614/TH neurons were scattered within the three parts of LDTg. PRV-614/TPH and PRV-614/TH neurons were located predominantly in the caudal of PPTg (cPPTg). These data provided direct neuroanatomical foundation for the identification of serotonergic and catecholaminergic projections from the mid-brain tegmentum to the kidney.

OBJECTIVE:To provide reference for further refinement and improvement of pharmaceutical administration in hospital accreditation standards in China.METHODS:According to Joint Commission International Accreditation Standards for Hospitals (6th edition) [called "JCI standards (6th edition)" for short] and Implementation Rules of Level 3 General Hospital Accreditation Standards in China (2011 edition) [called "Implementation Rules of Standards (2011 edition)" for short],the similarities and differences of pharmaceutical administration were studied and compared;the advantages and disadvantages of Implementation Rules of Standards (2011 edition) were analyzed to put forward some suggestions on this basis.RESULTS:The number of the items,standards and key points of pharmaceutical administration in JCI standards (6th edition) were 8,22,89,respectively.The number of above indexes in Implementation Rules of Standards(2011 edition) were 11,39,280,respectively.The similarities mainly reflected in relevant laws,regulations,rules and regulations to be followed in pharmaceutical administration,personnel requirements,drug procurement,reserve,storage and recall,drug dispensing and preparation,special drugs management,prescription management,antibiotics management,drug use monitoring,ADR reporting,etc.The specific regulation of JCI standards (6th edition) involved drug management system documentation review,drag acquisition at night or after pharmacy closed,drug list examination,comparison of drug use list before admission and the first medical order,etc.The specific regulation of Implementation Rules of Standards (2011 edition) involved the promotion of National Essential Medicine System and the construction of clinical pharmacist system.CONCLUSIONS:In China,Implementation Rules of Standards (2011 edition) have clearly defined the National Essential Medicine System and the construction of clinical pharmacist system,and are more in line with the development requirements of medical and health services in China at the present stage;but there are also some deficiencies compared to JCI standards (6th edition),including the revision update,the management of the patient's own medicine,the safety management of the patient's identity,measurability of the accreditation standards.Implementation Rules of Standards should be further refined and perfected by updating version regularly,strengthening the management of details,increasing measurability of the accreditation standards,strengthening drug management outside the department of pharmacy,enhancing drug supply management,etc.