Objective To construct a rat model of trigeminal neuralgia(TN)to explore the expression of high-mobility group box-1(HMGB1)in the trigeminal ganglion(TG)and the possible mechanism of HMGB1's effect on pain.Methods TN model was constructed by infraorbital nerve constriction and divided into operation group(CCI group)and Sham group,and the success of the model construction was determined through mechanical pain thresh-old assessment.Real time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect high mobility group protein B1(HMGB1),Toll receptor 4(TLR4),and Nuclear Factor Kappa B(NF-κB)mRNA and protein expression in the ipsilateral trigeminal ganglion(TG)of the Sham and CCI rats.50 mg/kg HMGB1 inhibi-tor glycyrrhizin(GL)was injected intraperitoneally every day for two weeks,and normal saline(NS)was used as control.The patients were divided into CCI group,CCI+NS group and CCI+GL group.HMGB1,TLR4,and NF-κB mRNA and protein expression in the ipsilateral trigeminal ganglion(TG)were detected by RT-qPCR and West-ern blot in CCI group,CCI+NS group,and CCI+GL group.Results The mechanical threshold on the operated side of the rat continued to decrease(P<0.05),and mechanical pain threshold identification model was success-fully constructed.After chronic compressive injury to the infraorbital nerve in rats,HMGB1,TLR4,and NF-κB mRNA and protein expression in TG on the operated side increased(P<0.05);After administration of HMGB1 inhibitor Glcyrrhizin,HMGB1,TLR4,NF-κB showed a decrease(P<0.05).Conclusion HMGB1 is associat-ed with TN,and HMGB1 may be involved in the pathogenesis of TN through TLR4/NF-κB signaling pathway.

Objective To investigate the pathological role of detect protein kinase C(PKC)/ transient receptor po-tential vanilloid subtype 1(TRPV1) pathway in trigeminal neuralgia (TN) in rats.Methods The infraorbital nerve-chronic constriction injury (ION-CCI) was used to establish a rat model of TN.The rats were randomly di-vided into Sham group,CCI group, CCI+DMSO group and CCI+GF109203X (a PKC inhibitor) group.The me-chanical pain threshold of the rats was measured using a Von Frey brush.qRT-PCR and Western blot were used to detect PKC and TRPV1 in the trigeminal ganglion (TG) .HE staining was used to observe the pathological changes of TG.Results The mechanical pain threshold significantly decreased (P <0.05), and the expression of phos-phorylated PKC(p-PKC) and TRPV1 in TG significantly increased in the CCI group (P<0.05).Histopathological results showed that compared with the Sham group,the CCI group observed significant changes in TG such as in-creased inflammatory cell infiltration and nerve cell swelling.Injection of GF109203X effectively reduced the phos-phorylation of PKC and the expression of TRPV1 in the TG of rats, and the mechanical pain threshold of the rats in-creased (P<0.05) .Under the light microscope, cell swelling and inflammatory cells in the TG were reduced.Conclusion PKC/TRPV1 pathway may be involved in trigeminal neuralgia in rats.

Objective : To explore the effects of an ultradian light cycle on the periodic expression of glutamate re⁃ ceptors in the suprachiasmatic nucleus and the lateral habenula nucleus. Methods : An ultradian light cycle T7 (3. 5 hours/3. 5 hours light : dark) was used to establish an ultradian light cycle model group and T24( 12 hours/ 12 hours light : dark) was used to establish control group. The expression of key proteins in the brain regions of the suprachiasmatic nucleus ( SCN) and the lateral habenula ( LHB) of the hypothalamus were analyzed by Western blot. Including glutaminergic receptors , pituitary adenylate cyclase activating polypeptide (PACAP) receptors , and downstream signaling molecules. Results : Western blot results showed that the expression of α⁃amino⁃3 ⁃hydroxy⁃5 ⁃methyl⁃4 ⁃isoxazole⁃propionic acid receptor subunit 2 (GluR2) in SCN under Zeitgeber time (ZT ) 1 and ZT 5 increased under an ultradian light cycle compared with normal photoperiod ( P < 0. 05 , P < 0. 01) , the overall expression of GluR2 in T7 group was higher than that in T24 group (P < 0. 01) , the overall expression of GluR2 in LHB group T7 was lower than that in T24 group (P < 0. 01) . Compared with T24 photoperiod , the overall expression of N ⁃methyl⁃D ⁃aspartic acid receptor subunit 2 (NR2B) in SCN increased under T7 photoperiod (P < 0. 05) , the overall expression of phosphorylation of extracellular signal⁃related kinase ( P⁃ERK) in LHB significantly increased under T7 photoperiod (P < 0. 05) . Conclusion An ultradian light cycle would cause an up⁃regulating of GluR2 and NR2B expression , a down⁃regulating of GluR2 expression and up⁃regulating P ⁃ERK expression in LHB.

Objective : To study the role of Toll⁃like receptor 7 (TLR7) in trigeminal nerve pain in SD rats , and to explore the mechanism of TLR7 mediating related inflammatory factors through the activation of NF⁃κB pathway during the process of pain. Methods : The rat model of trigeminal neuralgia (TN) was established by infraorbital nerve constriction injury(ION⁃CCI) . The expression of TLR7 in trigeminal ganglion (TG) was detected by qRT⁃PCR and Western blot. Hydroxychloroquine (HCQ) , an inhibitor of TLR7 , was given to ION⁃CCI rats by intragastric administration. The expression of TLR7 and its downstream signal pathway NF⁃κB subunit p65 , p ⁃p65 and inflammatory factors (TNF⁃α , IL⁃1β) in TG were detected. Results : After trigeminal nerve injury induced by ligation of infraorbital nerve , the expression of TLR7 in TG of rats significantly increased. After administration of TLR7 inhibitor, the expression of TLR7 , TNF⁃α and IL⁃1β in TG decreased , the translocation and phosphorylation of p65 nucleus decreased , the activation of NF⁃κB signal pathway was inhibited , and the mechanical pain induced by ION⁃CCI was relieved in male SD rats. Conclusion TLR7 in TG regulates neuropathic pain by activating NF⁃κB signal pathway in primary sensory neurons and mediating the expression of inflammatory cytokines TNF⁃α and IL⁃1β .

Many people affected by fragile X syndrome (FXS) and autism spectrum disorders have sensory processing deficits, such as hypersensitivity to auditory, tactile, and visual stimuli. Like FXS in humans, loss of Fmr1 in rodents also cause sensory, behavioral, and cognitive deficits. However, the neural mechanisms underlying sensory impairment, especially vision impairment, remain unclear. It remains elusive whether the visual processing deficits originate from corrupted inputs, impaired perception in the primary sensory cortex, or altered integration in the higher cortex, and there is no effective treatment. In this study, we used a genetic knockout mouse model (Fmr1^KO), in vivo imaging, and behavioral measurements to show that the loss of Fmr1 impaired signal processing in the primary visual cortex (V1). Specifically, Fmr1^KO mice showed enhanced responses to low-intensity stimuli but normal responses to high-intensity stimuli. This abnormality was accompanied by enhancements in local network connectivity in V1 microcircuits and increased dendritic complexity of V1 neurons. These effects were ameliorated by the acute application of GABA_A receptor activators, which enhanced the activity of inhibitory neurons, or by reintroducing Fmr1 gene expression in knockout V1 neurons in both juvenile and young-adult mice. Overall, V1 plays an important role in the visual abnormalities of Fmr1^KO mice and it could be possible to rescue the sensory disturbances in developed FXS and autism patients.
Animals ; Disease Models, Animal ; Fragile X Mental Retardation Protein/metabolism* ; Fragile X Syndrome/metabolism* ; Humans ; Mice ; Mice, Knockout ; Neurons/metabolism*

Objective : To investigate the co-labeling of c-Fos protein expression and oxytocin ( OXT) neurons in related brain regions in mice injected with dexmedetomidine( Dex) by intraperitoneal injection． Methods : 8 －10 week old C57 mice were divided into Saline group and Dex group．The Saline group was intraperitoneally injected with normal saline，the Dex group was intraperitoneally injected with Dex ( 100 μg / kg) ，the control group was giv- en normal saline (Saline group) ，and the experimental group was given Dex 100 μg / kg (Dex group) ．120 minutes after intraperitoneal injection，the brains of two groups of mice were collected by perfusion，and thenfrozen section and immunofluorescence microscopy imagingwere performed. Results : Immunofluorescence c-Fos protein expres- sion in Dex group and Saline group．The results showed that compared with the Saline group，the Dex group had higher thalamotomy in the supraoptic nucleus ( SON) ，ventrolateral preoptic nucleus ( VLPO) ，subfornical organ (SFO) ，solitary tract (SOL) and area postrema (AP) ，and the difference was statistically significant．C-Fos in the locus coeruleus(LC) ，lateral hypothalamic area (LH) ，laterodorsal tegmental nucleus (LDT) ，parabigeminal nu- cleus (PB) ，ventral tuberomammillary nucleus ( TMN) and motor cortex M1 / M2 protein expression decreased， and the difference was statistically significant (P＜0. 05) ．There was no difference in c-Fos protein expression in parathalamic nucleus (PVN) and suprachiasmatic nucleus ( SCN) ．The results of the co-labeling group of OXT neurons and c-Fos protein in the PVN and SON brain regions showed that : compared with the Saline group，there was no significant difference in the co-labeling of OXT neurons and c-Fos protein in the PVN and SON brain regions of the Dex group． Conclusion The immunofluorescence co-labeling experiment shows that Dex can activate sleep- promoting and other related brain regions，and has a certain inhibitory effect on wake-promoting and exercise-relat- ed brain regions，however，it does not affect the co-labeling of c-Fos protein and OXT neurons in PVN and SON brain regions.

Objective To analyze the epidemiological features,spectrum and case fatality of malignant tumor patients in Shenzhen city,to provide evidence for the development of prevention and treatment strategies on malignant tumor in Shenzhen.Methods All the hospitalized malignant tumor patients including deaths,were monitored from 1995 to 2014 in Shenzhen,and data was analyzed by SPSS 20.0 software.Results There were 160 988 inpatients of malignant tumors between 1995 and 2014 in Shenzhen.The top three hospitalized tumors were lung (13.64％),liver (11.13％) and breast (7.86％) cancers.Numbers of the malignant tumor inpatients had been rapidly increasing during the past 20 years,12.3 times in 2014 higher than in 1995.The total number of deaths due to malignant tumors was 19 460.Deaths of the top three malignant tumors were lung (24.40％),liver (19.84％) and colorectal (8.63％) cancers and the number of deaths was increasing,12.5 times higher in 2014 than in 1995.The overall case fatality rate was 12.09％.The annual percent change (APC) of malignant tumors case fatality rate was 9.7％(95％CI:2.0％-18.0％),during 1995-2003,with an increasing trend (t=2.72,P＜0.05).The APC of case fatality rate during 2003-2014 was-3.4％(95％CI:-7.6％-1.1％),but the decreasing trend (t=-1.63,P＞0.05) was not statistically significant.The top three major malignant tumors related to case fatality rate were lung cancer (21.62％),liver cancer (21.39％),and esophageal cancer (16.50％).The case fatality rates of leukemia and liver cancer had decreased during the past 20 years.The case fatality rates of cancers in lung,esophagus,stomach,breast,colorectal and nasopharyngeal,had all increased.The number of male patients was significantly exceeding the females (x2=41.691,P＜0.01),with sex ratio as 1.65:1.From age 35 and on,the number of deaths due to malignant tumors increased significantly,with the peak after 60 years of age.Conclusions The number of malignant tumor inpatients had an annual increase as well as the case fatality rate.Cancers in lung,liver appeared the leading causes of death among the malignant tumor patients,with elderly in particular.Strategies related to the prevention and treatment of cancers in lung,liver should be strengthened.

Objective To investigate the way of fear conditioning memory model evoked and erased by foot-shock in tree shrew. Methods First, detect the tree shrew activities regularly in light/dark box. Second, test a suitable voltage degree of foot shock on tree shrew. Third, investigate the memory formation and erasing of fear conditioning on tree shrew of trial group. Results The duration of tree shrew (n=4) stay in the dark-box was significantly lon-ger than that of in the light box (P<0. 01) in normal condition. In the same environment of two light boxes, given different voltage degrees, the durations of tree shrew (n=6) stay in the stimulating chamber gradually reduced and the durations of tree shrew stay had significant difference between stimulatus chamber and no stimulatus chamber when the stimulus voltage up to 12 V ( P<0. 05 ) , 16 V ( P<0. 01 ) and 20 V ( P<0. 01 ) . The animal of trial group ( n=4 ) could build up the fear conditioning memory of the dark box with the stimulus of 16 V foot-shock in the dark box ( P<0. 001 ) . After formation of the fear conditioning memory, the same stimulus in light box ap-peared for 4 days. The durations of tree shrew stay in trial group (n=4) decreased in light box, and there was no significant difference between the trial group and the control group. Conclusion Tree shrew prefers to stay in the dark box. The suitable voltage for foot-shock on tree shrew is 16 V. The fear conditioning memory can be evoked and erased by foot-shock.

Background:Visceral hypersensitivity is considered to be one of the major pathophysiological mechanisms of irritable bowel syndrome. Aims:To investigate the expression of TRPV1 and electrophysiological characteristics of colon-specific dorsal root ganglion( DRG)neurons in rat model of visceral hypersensitivity. Methods:Twenty 10-day-old rats were randomly divided into two groups. In model group,visceral hypersensitivity was induced by colorectal administration of acetic acid;while in control group the same amount of saline was administered. Colon-specific DRG neurons were labeled retrogradely by injection of DiI,a fluorochrome,into the colon wall. Expression of TRPV1 in DRG neurons was detected by immunofluorescence and the electrophysiological characteristics of DRG neurons was detected by using patch-clamp technique. Results:In model group,the expression rate of TRPV1 in colon-specific DRG neurons was significantly higher than that in controls(46. 1% vs. 36. 6% ,P <0. 01),the average rheobase was significantly decreased[(57. 80 ±1. 32)pA vs.(73. 45 ± 4. 51)pA,P < 0. 05],while the frequency of action potentials(APs)in response to doubling rheobase stimulation was significantly increased[(8. 20 ± 1. 10)Hz vs. (4. 54 ± 0. 66)Hz,P < 0. 05]. Score of abdominal withdrawal reflex(AWR)under a 60 mm Hg colorectal distention was positively correlated with the expression rate of TRPV1 and the frequency of APs in response to doubling rheobase stimulation(r = 0. 87 and r = 0. 73,P < 0. 01),but was negatively correlated with the rheobase(r = - 0. 81,P < 0. 01)in model group. Conclusions:Increased expression of TRPV1 and excitability in colon-specific DRG neurons might be a crucial step in formation of visceral hypersensitivity.

Objective To explore whether gap junction disturbances are involved in the pathogenesis of levodopa-induced dyskinesia ( LID ). Methods The hemi-parkinsonian ( PD ) rat was treated intraperitoneally with L-dopa methylester (20 mg/kg) and benserazid (10 mg/kg) for 21 days and abnormal involuntary movement was evaluated to establish LID rat model. The experimental animals were divided into three groups: LID group, PD group and normal control group, respectively. The behavior responses of intraperitoneal injection of different doses of carbenoxolon and intracerebroventricular injection of quinine were observed to estimate the effects of gap junctional blockade on the abnormal involuntary movement ( AIM ) in the rat model of LID. Double immunofluorescence labeling was used to analyze the expression of connexin 36 ( Cx36 ) in enkephalin positive medium spiny neurons and parvalbumin ( PV ) positive interneurons in the striatum. Western blottings was used to observe the expression of Cx36 in the striatum and moter cortex. Results Behavioral characteristics indicated that high dose of carbenoxolone ( >60 mg/kg) intraperitoneal injection and intracerebroventricular injection of quinine ( 0.5, 1.0, 2.0 μmol/L, > 2.5 μmol/L ) could decrease the AIM score of LID rats. Western blotting indicated that expression of Cx36 in lesioned striatum and motor cortex of LID rat model was 219.56% ±18.12% and 226.03% ±16.33%, respectively, which induced a significant upregulation in comparison with the normal control group (104.05% ±3.82%, t=15.389, P<0.01;105.27% ±2.82%,t=8.074, P<0.01) and untreated PD group (119.31% ±8.92%, t=13.356, P<0.01; 138.20% ±17.88%, t=5.872, P<0.01). Double immunofluorescence labeling staining revealed that Cx36 expression was increased in Enk-positive striatum neurons in LID model ( 57.59% ±5.36%) compared with that in normal control group (32.67% ±4.22%) and PD group (37.24% ±0.86%, F=78.060, P<0.01). The expression of Cx36 in PV-positive interneurons was also elevated in LID group (68.49% ±11.60%) in comparison with normal control group ( 40.43% ± 2.30%) and PD group ( 31.92% ± 5.68%, F = 39.567, P < 0.01 ).Conclusions The Cx36 expression is generally increased in lesioned striatum and motor cortex of LID rat model. In the striatum, the up-regulation of Cx36 is specifically observed in Enk-positive striatum neurons and in PV-positive interneurons. The dyskinesia behavior of LID rats can be significantly reduced by treatment with gap junction blockade. All these results suggest that gap junction dysfunction may play an important role in the pathogenesis of LID.