The thalamocortical (TC) circuit is closely associated with pain processing. The hyperpolarization-activated cyclic nucleotide-gated (HCN) 2 channel is predominantly expressed in the ventral posterolateral thalamus (VPL) that has been shown to mediate neuropathic pain. However, the role of VPL HCN2 in modulating TC circuit activity is largely unknown. Here, by using optogenetics, neuronal tracing, electrophysiological recordings, and virus knockdown strategies, we showed that the activation of VPL TC neurons potentiates excitatory synaptic transmission to the hindlimb region of the primary somatosensory cortex (S1HL) as well as mechanical hypersensitivity following spared nerve injury (SNI)-induced neuropathic pain in mice. Either pharmacological blockade or virus knockdown of HCN2 (shRNA-Hcn2) in the VPL was sufficient to alleviate SNI-induced hyperalgesia. Moreover, shRNA-Hcn2 decreased the excitability of TC neurons and synaptic transmission of the VPL-S1HL circuit. Together, our studies provide a novel mechanism by which HCN2 enhances the excitability of the TC circuit to facilitate neuropathic pain.
Animals ; Mice ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/genetics* ; Neuralgia ; RNA, Small Interfering ; Thalamus/metabolism* ; Up-Regulation

OBJECTIVE: To investigate the effect of miR-133b on cardiomyocyte apoptosis induced by myocardial ischemia-reperfusion (I/R) and explore the mechanism. METHODS: Thirty-six adult SD rats were randomized into sham-operated group, I/R group, AdmiR-NC group and AdmiR-133b group, and rat models of myocardial I/R were established in the latter 3 groups with myocardial injections of saline or recombinant adenoviruses in the left ventricle. The expression of MiR-133b was detected using RT-qPCR, and cardiac function of the rats was determined using FDP 1 HRV and BRS analysis system. Serum CK-MB and cTnI levels were determined by ELISA, myocardial injury was evaluated with HE staining, cardiomocyte apoptosis was detected by flow cytometry, and ROS content was determined using a DCFH-DA probe. In the in vitro experiment, H9C2 myocardial cells with hypoxia/reoxygenation (H/R) treatment were transfected with Mir-NC or MiR-133b mimic, and the cellular expression of MiR-133b, cell apoptosis, and ROS content were determined. Dual luciferase reporter assay was performed to verify the targeting relationship between miR-133b and YES1. The effects of pc-YES1 or miR-133b mimic transfection on YES1 expression, apoptosis, and ROS content in H9C2 cells were evaluated. RESULTS: Compared with those in I/R group, miR-133b expression was obviously up-regulated, LVEDP, cTnI and CK-MB levels were significantly decreased, and LVSP, +dp/dt, -dp/dt, HR and CF levels were increased in admiR-133b group ( CONCLUSIONS miR-133b can inhibit I/R-induced myocardial cell apoptosis and ROS accumulation by targeting YES1 to reduce myocardial I/R injury in rats.
Animals ; Apoptosis ; MicroRNAs/genetics* ; Myocardial Reperfusion Injury ; Myocytes, Cardiac ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species

Objective:To investigate the specificity and sensitivity of four methods including ultrafastreal-time fluorescence polymerase chain reaction (PCR), real-time fluorescence (RT)-PCR, enzyme-linked immunosorbent assay (ELISA) and gold immunochromatography assay (GICA) for the detection of novel Bunya virus, so as to provide experimental basis for the early diagnosis of severe fever with thrombocytopenia syndrome (SFTS).Methods:Serum samples from 86 clinically diagnosis SFTS patients admitted to the Jinan Infectious Diseases Hospital Affiliated to Shandoug University were tested by ultrafast real-time fluorescence PCR, RT-PCR, ELISA and GICA during June 1 to September 30, 2017. Chi-square test was used for statistical analysis.Results:Among 86 serum samples, the positive rate of novel Bunya virus of ultrafast real-time fluorescence PCR, RT-PCR, IgM-ELISA, IgG-ELISA, IgM-GICA and IgG-GICA were 82(95.34%), 79(91.86%), 41(47.67%), 8(9.3%), 19(22.09%) and 3(3.49%), respectively. The specificity of ultrafast real-time fluorescence PCR was 100%, and the sensitivity was 1×10 3 copies/mL.Repeated amplification test showed that the variation coefficient of the computed tomography value was <2%.During phases one, two and three, the positive rates of ultrafast real-time fluorescence PCR were 41(97.62%), 34(94.44%) and 7(87.50%), and RT-PCR were 39(92.86%), 33(91.67%) and 7(87.50%), respectively. During phases one and two, the positive rate of ultrafast real-time fluorescence PCR was slightly higher. The positive rate of anti-novel Bunya virus antibody (IgM) tested by ELISA had a significant increase from phase one (28.57%)to phase three (87.50%). There were statistical differences between phase two and phase, as well as between phase three and phase one ( χ2=8.347 and 7.561, respectively, both P<0.01). IgM-GICA also had an increase from phase one (14.29%) to phase two (33.33%)( χ2=3.962, P<0.01), while it was still lower than the other tests.In phase one, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG)( χ2=33.740, 55.080, 49.010 and 64.340, respectively, all P<0.01). In phase two, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG) ( χ2=7.700, 46.720, 23.700 and 50.630, respectively, all P<0.01). In phase three, the positive rates of ultrafast real-time fluorescence PCR, RT-PCR and IgM-ELISA were equivalent, which were all higher than those of IgG-ELISA and GICA (both IgM and IgG). The positive rates of RT-PCR and IgG-ELISA, IgM-GICA and IgG-GICA were significantly different (all χ2=6.250, all P<0.05). Conclusion:In the early detection of novel Bunya virus, ultrafast real-time fluorescence PCR has higher sensitivity, specificity, good repeatability and high stability, which greatly reduces the amplification time compared with the traditional RT-PCR, and is of great value in the early and rapid diagnosis of SFTS.

Objective To observe the effects of lactobacillus complex capsules in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and different degrees anorexia. Methods 100 anorexia score 1 and 100 anorexia score 2-5 AECOPD patients were divided into 4 groups: the therapy group 1 (anorexia score 1,n = 50) and the therapy group 2 (anorexia score 2-5,n = 50) with conventional and lactobacillus complex capsules therapy, the control group 1 (anorexia score 1, n = 50) and the control group 2 (anorexia score 2-5,n = 50) with conventional therapy. Parameters were recorded, which include length of hospital stay , markers of inflammation , arterial blood gases , lung function , clinical symptoms , death rate and side-effects of lactobacillus complex capsules. Results After treatment, WBC, trophil ratio, length of hospital stay in the therapy group 2 were lower than the control group 2(P < 0.05). The PCO2 in the therapy group 1 was lower than the control group 1 (P < 0.05). There were no differences in the rest parameters between the two control groups and the two therapy groups (P > 0.05). No side-effects like rashes, diarrhea, were observed in the two therapy groups. Conclusion For anorexia patients with AECOPD, oral lactobacillus complex capsules can safely control infection and reduce length of hospital stay.

[ ABSTRACT] Rebound depolarization is a special phenomenon of the neurons which generates action potential fol-lowed by a hyperpolarization stimulation.It can be recorded in many kinds of neurons and is the intrinsic membrane charac-teristic of them.Rebound depolarization plays an important role in regulating the firing pattern, rhythmic activity and sy-naptic plasticity of neurons.This review focuses on the basic characteristics, the function and mechanism of the rebound depolarization in physiological and pathological conditions, which provides reference for the clinical treatment of rebound depolarization-related diseases.

Objective To observe the effects of lactobacillus complex capsules in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods A hundred AECOPD patients were divided into 2 groups: the therapy group(n = 50) with conventional and lactobacillus complex capsules therapy, and the control group with conventional therapy only. Parameters before and after therapy were compared, which included length of hospital stay, arterial blood gases, dyspnea grade, respiratory function test, markers of inflammation, anorexia score, diarrhea, fungi infection, breathing rate, death rate and side-effects of lactobacillus complex capsules. Results After 7 days of treatment, the blood leukocyte count and neutrophil ratio in the therapy group were lower than that of the control group (P < 0.05). The rate of anorexia score ≤ 2 in the therapy group was higher than that of the control group (P < 0.05). There were no differences in the rest parameters among the two groups(P > 0.05). There were no side-effects, such as skin rash and diarrhea, to be observed in the therapy group. Conclusion Oral lactobacillus complex capsule can be safe, anti-infective and improve appetite in patients with AECOPD.

OBJECTIVETo investigate the effect of minocycline on hyperpolarization-activated current (Ih) in the substantia gelatinosa (SG) neurons in rat spinal dorsal horn.

METHODSIn vitro spinal cord transverse slices were prepared from 3-5-week-old male Sprague-Dawley rats. Using whole-cell patch clamp technique, Ih currents were recorded before and after bath application of minocycline (1-300 µmol/L) to the SG neurons.

RESULTSIh currents were observed in nearly 50% of the recorded neurons, and were blocked by Ih blocker CsCl and ZD7288. Minocycline rapidly and reversibly reduced the amplitude of Ih and decreased the current density in a concentration-dependent manner with an IC50 of 34 µmol/L.

CONCLUSIONMinocycline suppresses the excitability of SG neurons through inhibiting the amplitude and current density of Ih and thereby contributes to pain modulation.

Animals ; Male ; Minocycline ; pharmacology ; Neurons ; drug effects ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Substantia Gelatinosa ; cytology

Objective To observe the efficacy and safety of radiofrequency thermocoagulation of lumbar sympathetic ganglia combined with pregabalin in the treatment of complex regional pain syndrome(CRPS).Methods 26 patients with lower limb CRPS were selected and treated by the radiofrequency thermocoagulation of lumbar sympathetic ganglia combined with oral pregabalin capsule.The visual analogue scale (VAS)and the quality of sleep(QS)were adopted to evaluate the pain change before treatment and on 1,7,14,28,56 d after treatment.The temperature change of lower limb skin and the occurrence situation of adverse reactions were recorded.Results Compared with before treatment,the scores of VAS and QS at different time points after treatment were decreased significantly (P <0.05),the skin temperature of affected lower limb after treatment was increased significantly (P <0.05).The total effective rates on 28,56 d after treatment were 88.46% and 96.15% respectively.The adverse reactions were mainly dizziness and somnolence.No severe complications such as vascular,neural and intra-abdominal organs injury were found in the treatment process.Conclusion The radiofrequency thermocoagulation of lumbar sympathetic ganglia combined with pregabalin in the treatment of CRPS can rapidly alleviate pain,improve the patients′quality of sleep and living.

Objective To compare the difference of the positive rates between real-time RT-PCR and ELISA-IgM methods in diagnosis of severe fever with thrombocytopenia syndrome (SFTS) cases.Method 138 serum samples from 78 suspected acute SFTS patients were collected continuously in our hospital,2014.And 91 serum samples from 49 confirmed cases were detected by real-time RT-PCR and ELISA-IgM,simultaneously.The results were analyzed by Software SPSS17.0.Result 138 serum samples were detected by the real-time RT-PCR and ELISA-IgM methods,and the positive rate was 51.45％,43.48％ respectively.There was significant difference between their positive rates (P ＜ 0.05).The courses of disease influenced the positive rates notablely in 49 confirmed cases between the real-time RT-PCR and ELISA-IgM assays.When the serum samples were collected within 8 days onset of illness to be detected,the positive rate of real-time RT-PCR method was higher than that of ELISA-IgM method.While the positive rate of ELISA-IgM showed a rising tendency along with the course extension.Conclusion Real-time RT-PCR would be the first choice for the early diagnosis of suspected SFTS cases.While ELISA-IgM methods could be the necessary supplemental assay and it would be the important diagnostic method as the extension of the course of SFTS.

Objective To evaluate the clinical significance of carcinoembryonic antigen (CEA),cancer antigen 125 (CA125),carbohydrate antigen 19-9 (CA19-9) and eytokeratin 21-1 fragment (CYFRA21-1) assays of sputum in patients with lung cancer. Method Fifty-two cases with lung cancer and 46 cases with benign lung diseases underwent detection of CEA ,CA125 ,CA19-9 and CYFRA21-1 in sputum by using the method of chemi-luminescent enzyme immunoassay. Results The levels of CEA,CA125,CA19-9 and CYFRA21-1 in sputum of 52 cases with lung cancer were (27.6±31.2) μg/L, (76.4±65.2)kU/L, (56.1±31.6) kU/L and ( 25.2±9.1 )μg/L respectively. But the levels of those of 46 cases with benign lung diseases were (6.1±7.5)μg/L, (23.7±7.9) kU/L, (17.3±10.2) kU/L and (1.2±1.7)μg/Lrespectively. The levels of these tumor markers in sputum in patients with lung cancer were significantly higher than those in patients with benign lung diseases(P< 0.01 ). The diagnostic sensitivity of CEA,CA125,CA 19-9 and CYFRA21-1 in sputum in 52 cases with lung cancer was 42.3%( 22/52 ), 46.2% (24/52), 36.5%( 19/52 ) and 51.9% (27/52) respectively ( P > 0.05 ). Among the cancer patients, the sensitivity of sputum CEA in patients with adenocareinoma was significantly higher than that in patients with squamons cell carcinoma (X2= 4.193, P < 0.05 ) ; while the sensitivity of sputum CYFRA21-1 in patients with squamous cell carcinoma was significantly higher than that in patients with adenocarcinoma ( X2 = 4.806,P < 0.05 ). The sensitivity of CA125 in advanced lung cancer( Ⅲ +Ⅳ stage) was higher than that in early lung cancer( Ⅰ+Ⅱstage) (X2= 5.202,P < 0.05 ). Compared with the single tumor marker assaying, the combination of CEA,CA 125, CA 19-9 and CYFRA21-1 in sputum could significantly improve the sensitivity and accurate value in the diagnosis of lung cancer (P<0.05). Conclusion To assay CEA,CA125,CA1g-9 and CYFRA21-1 in sputum is valuable in diagnosis of lung cancer, and the combination of CEA,CA125,CA19-9 and CYFRA21-1 in sputum can significantly improve sensitivity and accurate value in the diagnosis of lung cancer.