Involvement of Opioid Peptides in the Analgesic Effect of Spinal Cord Stimulation in a Rat Model of Neuropathic Pain / 神经科学通报·英文版

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.

Responses of Primary Afferent Fibers to Acupuncture-Like Peripheral Stimulation at Different Frequencies: Characterization by Single-Unit Recording in Rats / 神经科学通报·英文版

The pain-relieving effect of acupuncture is known to involve primary afferent nerves (PANs) via their roles in signal transmission to the CNS. Using single-unit recording in rats, we characterized the generation and transmission of electrical signals in Aβ and Aδ fibers induced by acupuncture-like stimuli. Acupuncture-like signals were elicited in PANs using three techniques: manual acupuncture (MAc), emulated acupuncture (EAc), and electro-acupuncture (EA)-like peripheral electrical stimulation (PES). The discharges evoked by MAc and EAc were mostly in a burst pattern with average intra-burst and inter-burst firing rates of 90 Hz and 2 Hz, respectively. The frequency of discharges in PANs was correlated with the frequency of PES. The highest discharge frequency was 246 Hz in Aβ fibers and 180 Hz in Aδ fibers. Therefore, EA in a dense-disperse mode (at alternating frequency between 2 Hz and 15 Hz or between 2 Hz and 100 Hz) best mimics MAc. Frequencies of EA output >250 Hz appear to be obsolete for pain relief.

Development of an Autism Subtyping Questionnaire Based on Social Behaviors / 神经科学通报·英文版

Autism spectrum disorder can be differentiated into three subtypes (aloof, passive, and active-but-odd) based on social behaviors according to the Wing Subgroups Questionnaire (WSQ). However, the correlations between the scores on some individual items and the total score are poor. In the present study, we translated the WSQ into Chinese, modified it, validated it in autistic and typically-developing Chinese children, and renamed it the Beijing Autism Subtyping Questionnaire (BASQ). Our results demonstrated that the BASQ had improved validity and reliability, and differentiated autistic children into these three subtypes more precisely. We noted that the autistic symptoms tended to be severe in the aloof, moderate in the passive, and mild in the active-but-odd subtypes. The modified questionnaire may facilitate etiological studies and the selection of therapeutic regimes.

Transcutaneous electrical acupoint stimulation for asthenozoospermia / 中华男科学杂志

Objective@#To study the effect of transcutaneous electrical acupoint stimulation (TEAS) in the treatment of asthenozoospermia.@*METHODS@#We randomly divided 72 asthenozoospermia patients into a 2 Hz TEAS (n = 29), a 100 Hz TEAS (n = 20), and a blank control group (n = 23), those in the former two groups treated by 30 minutes of TEAS at 2 Hz and 100 Hz respectively, applied to the acupoints of bilateral Shenshu, left Zusanli, and Guanyuan, once a day for 60 days, while those in the blank control group left untreated. Using computer-assisted sperm analysis (CASA), we examined sperm concentration and motility as well as the percentages of grade a and grade a+b sperm in different groups of the patients.@*RESULTS@#Compared with the baseline, 2 Hz TEAS significantly increased sperm motility (［12.76 ± 1.39］ vs ［18.89 ± 2.46］%, P<0.05) and the percentage of grade a+b sperm ( ［10.68 ± 1.22］ vs ［16.32 ± 2.10］%, P<0.05) in the asthenozoospermic patients, while 100 Hz TEAS improved not only sperm motility (［12.32 ± 2.21］ vs ［23.81 ± 3.42］%, P<0.01) and the percentage of grade a+b sperm (［10.45 ± 1.98］ vs ［20.25 ± 2.82 ］%, P<0.01), but also the percentage of grade a sperm (［6.44 ± 1.16］ vs ［13.31 ± 2.30］%, P<0.05). Moreover, in comparison with the blank control group, 2 Hz TEAS also remarkably increased sperm motility (［9.57 ± 1.60］ vs ［18.89 ± 2.46］%, P<0.05) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［16.32 ± 2.10］%, P<0.05) in the asthenozoosperma patients, while 100 Hz TEAS improved not only sperm motility (［9.57 ± 1.60］ vs ［23.81 ± 3.42］%, P<0.01) and the percentage of grade a+b sperm (［7.81 ± 1.31］ vs ［20.25 ± 2.82］%, P<0.01) but also the percentage of grade a sperm (［4.87 ± 1.01］ vs ［13.31 ± 2.30］%, P<0.01). Meanwhile, the rate of clinical effectiveness was significantly higher in the 100 Hz TEASthan in the blank control group either in intention-to-treat (ITT) analysis (100% vs 18.18%) orper-protocol (PP) analysis (90% vs 0%), and so was it than in the 2 Hz TEAS group based on the data of ITT (100% vs 33.33%).@*CONCLUSIONS@#Both 2 Hz and 100 Hz TEAS are effective for the treatment of asthenozoospermia by improving sperm motility and vitality.

Effect of Itk down regulation on cytokines production in Jurkat cell / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the effect of Itk down regulation on Jurkat cell proliferation and inflammatory cytokines production, and provide useful data for Itk as an attractive target for potential drugs.</p><p><b>METHODS</b>Three shRNAs against different region of Itk were constructed and cotransfected with pEGFP-C1-hItk. The shRNA, which can knock down Itk, was selected and packed into lentivirus. After Jurkat cells were transfected with shRNA lentivirus, the change of Itk protein expression, cell proliferation and cytokines production was observed.</p><p><b>RESULTS</b>Itk mRNA was reduced about 55% in Jurkat cells transfected with Itk-shRNA1, compared with that in control cells shRNAnon (P < 0.05). Knocking down Itk expression had a profound inhibitory effect on Jurkat cell proliferation. In addition, there was a substantial decrease in level of cytokines, such as IL-2, IL-5, IL-10 and IFN-gamma, produced by cell transfected with Itk-shRNA1.</p><p><b>CONCLUSION</b>Knocking down Itk expression can inhibit Jurkat cell proliferation and inflammatory cytokines production.</p>

Research of ITK regulation on mouse spleen lymphocytes proliferation and differentiation / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>By using the RNAi method to inhibit Itk protein expression specificity, to observe lymphocytes proliferation and cytokines production, verify its function as a drug target.</p><p><b>METHODS</b>Designed siRNA aims at Itk sequence according to its sequence and solid structure, then electrotransfected into mouse spleen lymphocytes, We validated the decrease of Itk protein by Western-Blot, and detected the change of the cell proliferation by MTS and the change of inflammatory cytokines by ELISA.</p><p><b>RESULTS</b>Itk protein can be suppressed by Itk-siRNA, there were significantly reduced compared to its control group on cell proliferation as well as cytokine secretion such as IL-2, IL-4, IL-5, IFN-gamma. They all have statistical difference (P < 0.05).</p><p><b>CONCLUSION</b>Itk has an important immunomodulatory effect in mouse spleen lymphocytes proliferation and secretion of inflammatory cytokines.This can supply an experimental basis to regard Itk as drug target for inflammation therapy.</p>

Electroacupuncture analgesia is enhanced in transgenic nociceptin/orphanin FQ knock-out mice / 北京大学学报(医学版)

Objective: The modulatory effects of nociceptin/orphanin FQ ( OFQ) on acupuncture ( EA, a odern version of acupuncture)-induced analgesia are still controversial. Transgenic OFQ knock-out mice rovide us a useful tool to investigate the role of endogenous OFQ in EA analgesia. The present tudy aims to investigate the role of OFQ in the EA-induced analgesia with OFQ knock-out mice. ethods; Acupoints were selected as "Zusanli" (ST36) and "Sanyinjiao" (SP6) , EA parameters were as ollows; constant current output, rectangular (square) wave pulses, increased intensities of 0. 5, 0. 7,0. 9 mA with stepwise fashion, 10 min for each intensity, frequency of 100 Hz (with 0. 2 ms as ulse width) or 2 Hz ( with 0. 6 ms as pulse width). Tail flick latency (TFL) evoked by radiant heat was used to evaluate the change of pain threshold before, during and after EA application. Thenock-out mice were divided randomly into 3 groups; needling control, EA at 100 Hz and 2 Hz. ild-type mice of litter-mates of were used as Control. Results: It was found that OFQ knock-out mice had a longer basal ther-mal threshold; EA had enhanced analgesic effect in the knock-out mice than in wild-type Control mice. Conclusion: Endogenous OFQ might be algesic in basal condition and antagonize EA analgesia.

Intracellular clearance of Coxsakievirus B3 infection by short interfering RNA and its mechanism study / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the possibility of short interfering RNA (siRNA) inhibiting Coxsackievirus B3 (CVB3) infection in vitro, and discover the mechanism initially.</p><p><b>METHODS</b>We obtained proper effective dosage of siRNA by observing cytopathic effect (CPE). Estimate its antiviral activities and its pathway of siRNA by Western Blot assay and RT-PCR.</p><p><b>RESULTS</b>Results showed that siRNA-3753 can be effectively transfected into HeLa cells, we can achieve a high transfection efficiency up to 98.77% and its effect can last for 48 h stably in cells. 0.6 micromol/L siRNA-3753 got a high inhibiting effect of virus and didn't show any toxicity to cells. So we consider this concentration as the experimental concentration. siRNA-3753 can debase virus reproduction. The antiviral effect is sequence-specific and is not attributable to either interferon or the interferon response effectors protein kinase R (PKR).</p><p><b>CONCLUSION</b>The data confirmed that siRNA can effectively inhibit CVB3 infection in vitro, its antivirus effect was gained from specific debase of virus genome.</p>

Study on inhibition of coxsackievirus B3 infection in HeLa cell by short interfering RNA targeting 2B protein / 病毒学报

To study the inhibitory effect and function characteristics of small interfering RNA (siRNA) on cosxackievirus B3(CVB3) infection by RNA interference technique, siRNA-2B against the viral 2B region was synthesized and transfected into HeLa cell, which was then infected with CVB3. The efficiency of siRNA transfection was examined by FCM, the cell toxicity of siRNA-2B by MTT, and the antiviral ability of siRNA-2B by cytopathic effect (CPE), plaque reduction assay and RT-PCR. The results showed that siRNA-2B could be transfected efficiently into HeLa cell and lasted at least 48h. High concentration of siRNA-2B didn't show any sign of toxicity to cells. siRNA-2B exhibited a significant protective effect on cell viability by specific inhibition of viral replication. It showed a close relationship between the concentrations of siRNA-2B and the antiviral effects. siRNA-2B led to dramatical reduction of viral titers in supernatant of cell culture and weakened the reinfection ability of the virus. These data proposed that siRNA-2B, targeting 2B protein, can effectively inhibit CVB3 infection in HeLa cell and exhibits its transfection efficiency, viral inhibition specificity and adose-dependant manner, suggesting its potential role in prevention and treatement of CVB3 infection.

Short interfering RNA-mediated inhibition of coxsakievirus B3 infection in vitro / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate feasibility of inhibiting coxsackievirus B3 (CVB3) infection at cellular, protein and gene levels by using small interfering RNA (siRNA).</p><p><b>METHODS</b>Antiviral activities of siRNAs were evaluated by observing cytopathic effect (CPE), using plaque reduction Western blotting assays and RT-PCR.</p><p><b>RESULTS</b>Eight siRNAs were synthesized, among them, SiRNA-2, SiRNA-3, SiRNA-6 and SiRNA-7 which were targeted against sequences located in 2B, VP4, 2A and 3C section of CVB3 genome, were designed to have different effect of inhibiting CVB3 infection in vitro. SiRNA-2 showed the best protective effect, 95 percent inhibition of CVB3 cytopathic effect and plaque forming effect was observed at 0.0001 MOI, viral protein synthesis and replication were inhibited. SiRNA-2 showed 30 percent inhibition of virus at 0.1 MOI, 70 percent inhibition at 0.01 MOI, 88 percent inhibition at 0.001 MOI, and 99 percent inhibition at 0.0001 MOI 48 hours after CVB3 infection.</p><p><b>CONCLUSION</b>SiRNA could effectively inhibit CVB3 infection in vitro, siRNA-2, which is targeted against sequence in 2B section of CVB3 genome, seemed to be the best one among those synthesized in this study.</p>

Tramadol and dihydroetorphine produce synergistic analgesic effect and postpones acute opiate tolerance in rats / 生理学报

The present study investigated whether a co-application of tramadol (TRA) and dihydroetorphine (DHE) would exert a synergy in analgesic effect and delay acute tolerance development. Intraperitoneal injection of TRA (in mg) and subcutaneous injection of DHE (in ng) were delivered in fixed proportions (1:6.25, 1:12.5, 1:25, 1:50, 1:100, and 1:200). The effect of analgesia was accessed by tail-flick test and analyzed with isobolographic analysis. For test of acute tolerance, six successive injections of either TRA (20 mg/kg) alone, DHE (1 000 ng/kg) alone, or a combination of TRA (20 mg/kg) and DHE (250 ng/kg) were administered. We found that (1) except for 1 mg : 6.25 ng and 1 mg : 50 ng, combinations, all the other ratios produced a significant synergy in their analgesic effect; (2) the effect of analgesia induced by repeated TRA plus DHE injections lasted significantly longer, indicating a slower onset of acute tolerance. These results indicate that TRA and DHE injections in certain dose ratios can induce synergistic analgesia, which is resistant against the development of acute tolerance.

Protective effect of tripchlorolide on dopaminergic neurons in partially lesioned rat model of Parkinson's disease / 药学学报

<p><b>AIM</b>To study whether the immunosuppressant tripchlorolide (T4) exerts neuroprotective effect on dopaminergic neurons.</p><p><b>METHODS</b>A rat model of Parkinson's disease (PD) was set up by transection of the medial forebrain bundle (MFB) with a wire knife. The rotational behavior, HPLC-ECD, tyrosine hydroxylase (TH) immunocytochemistry, ELISA methods were used to evaluate the influence on the dopaminergic neurons following T4 treatment.</p><p><b>RESULTS</b>T4 treatment was shown to effectively attenuate the rotational behavior challenged by amphetamine (2.5 mg.kg-1, i.p.) in the PD rats. T4 markedly prevented the decrease of dopamine content in the striatum and the loss of dopaminergic neurons in the substantia nigra pars compacta. T4 was found to suppress the abnormal increase of TNF-alpha and IL-2 level in brain tissues of PD rats after MFB transection.</p><p><b>CONCLUSION</b>The evidence that the immunosuppressive Chinese herb extract T4 possesses neuroprotective activities on the dopaminergic neurons in PD rats was presented. The underlying mechanism of T4 may be relevant to its immunosuppressive activity.</p>