The effect of combining transcranial magnetic stimulation with escitalopram on neurological functioning, depression and serum inflammatory factors after a stroke / 中华物理医学与康复杂志

Objective:To investigate the effect of combining repeated high-frequency transcranial magnetic stimulation (rTMS) with escitalopram in treating the neurological functioning and post-stroke depression of stroke survivors.Methods:Eighty persons with post-stroke depression were randomly divided into an observation group and a control group, each of 40. The control group was treated with oral escitalopram, while the observation group also received transcranial magnetic stimulation at 5Hz. The magnetic stimulation intensity was 80% of each person′s resting motion threshold. The rTMS was administered once a day, 5 days a week for 8 weeks. Neurological functioning and depression were evaluated using National Institutes of Health stroke scoring (NIHSS) and the Hamilton depression scale before and after the 8 weeks of treatment. The levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2) and interleukin-6 (IL-6) in serum samples from the two groups were detected using enzyme-linked immunosorbent assays.Results:After treatment, the NIHSS and Hamilton scores and TNF-α, IL-2 and IL-6 levels in both groups had improved significantly, but in each case they were significantly better in the treated group, on average.Conclusion:Supplementing escitalopram with high-frequency transcranial magnetic stimulation more effectively improves depression and neurological functioning after a stroke.

Standing at a table while training improves the upper limb function of hemiplegic stroke survivors / 中华物理医学与康复杂志

Objective To explore the effect of standing at a table while training on the upper limb function and muscle surface electromyography of hemiplegics.Methods Sixty hemiplegic persons were randomly divided into an experimental group and a control group,each of 30.The affected upper limbs of the experimental group were trained while standing at a table,while the control group was trained while sitting.Before the treatment,as well as after 2 and 4 weeks of treatment,both groups' motor functioning was evaluated using the Fugl-Mayer upper limb assessment (FMA),as well as muscle surface electromyography.Results Before the treatment there was no significant difference betweenthe two groups' average FMA scores.After 2 and 4 weeks of treatment it had increased significantly in the experimental group,but in the control group the increase was not significant until the fourth week.In terms of surface myography,significant differences were observed in the biceps femoris and gastrocnemius muscles of both groups after 2 weeks.Two weeks later there was further significant improvement in both groups except for the tibialis anterior muscles of the control group.The differences between the two groups were significant after two weeks in the electromyograms of the biceps femoris,gastrocnemius muscle and anterior tibialis.After four weeks the differences between the groups in all of the electromyograms were significant.Conclusion Compared with the traditional sitting position,standing at a table while training can effectively improve the muscle activity of the rectus abdominis and the spine so as to promote the recovery of movement in hemiplegic upper limbs.

Effects of paired associative stimulation and repetitive transcranial magnetic stimulation on the excitability of the motor cortex / 中华物理医学与康复杂志

Objective To compare the effects of paired associative stimulation (PAS) and repetitive transcranial magnetic stimulation (rTMS) on motor cortex excitability.Methods The baseline corticospinal excitability of the left hemispheres of 10 healthy subjects was measured in terms of resting motor threshold (RMT) and other indicators of motor evoked potentials (MEP).On the following day they received PAS composed of trascranial magnetic stimulation (TMS) to the motor cortex of the left hemisphere and electric stimulation (ES) of the median nerve contralateral to the motor cortex,with an interval of 10ms between the TMS and ES (termed PAS10).The PAS10 was delivered at a frequency of 0.05 Hz and an intensity of 120％ of the RMT,for a total of 90 pulses.The MEP amplitude,MEP latency and RMT were evaluated one minute after the stimulation.After the PAS intervention,an interval of one week was allowed to eliminate any effect of PAS on motor cortex excitability.Then rTMS was delivered to the subjects' left motor cortex at the same time of day at a frequency of 1 Hz and an intensity of 120％ of the RMT,for a total of 1000 pulses.MEP amplitude,MEP latency and RMT were evaluated one minute after the stimulation.The two interventions were compared in terms of MEP amplitude,MEP latency and RMT.Results The average MEP amplitude,MEP latency and RMT at baseline were (2.93 ± 0.99) mV,(20.97 ± 1.67) ms,and (46.06 ±5.32) ％,respectively.One minute after PAS10,the MEP amplitude,MEP latency and RMT were (1.14 ± 0.76) mV,(21.87 ± 1.09) ms and (52.06 ±4.20) ％,respectively.One minute after rTMS,the MEP amplitude and latency and the RMT were (2.24 ± 0.79) mV,(20.88 ± 1.94) ms,and (49.00 ± 4.54) ％,respectively.The differences in MEP amplitude,MEP latency and RMT pre-and post-intervention were (0.69 ± 0.10) mV,(0.09 ±0.05) ms and (3.94 ± 0.93) ％,respectively for rTMS.For PAS10 they were (1.83 ± 0.14) mV,(0.90 ± 0.26)ms and (6.00 ± 1.13)％,respectively.The differences in MEP amplitude decrease and MEP latency lengthening between the two stimulation protocols were significant,but the difference in RMT elevation was not.Conclusion Both PAS10 and low frequency rTMS suppressed motor cortex excitability,but the suppressive effect of PAS10 is more significant.

The effects of paired associative stimulation on motor cortex excitability and the recovery of motor function after stroke / 中华物理医学与康复杂志

Objective To investigate the effects of paired associative stimulation (PAS) on the recovery of upper limb motor function in stroke patients,and to analyze the relationship between the change of motor cortex excitability in the contralesional hemisphere and the recovery of motor function in the affected upper limb.Methods Thirty hemiplegic stroke patients were divided randomly into a treatment group and a control group.Both groups were given routine rehabilitation therapy,but the treatment group also received PAS consisting of transcranial magnetic stimulation (TMS) of the intact motor cortex and electrical stimulation (ES) of the median nerve at the wrist of the intact arm with an interval of 10 ms between the TMS and ES (PAS10).The PAS10 was delivered at a frequency of 0.05 Hz and an intensity of 120％ the resting motor threshold (RMT),once daily for 30 minutes,five times a week for 4 weeks.Corticospinal excitability was measured using motor evoked potentials (MEP) and the RMT.The FuglMeyer upper limb assessment (FMA),Brunnstrom staging and the modified Barthel index (MBI) were also applied before and at the end of the 4 weeks of treatment.Correlation was sought between any changes in MEP amplitude,the RMT of the contralesional hemisphere and changes in the FMA results.Results Before the intervention there were no significant differences between the two groups in terms of any of the assessments.After 4 weeks of treatment,all the assessments had changed significantly compared to those before the treatment,but there were still no significant differences between the two groups in terms of any the assessments.After 4 weeks of treatment,the differences in MEP amplitude from the contra-lesional hemisphere and the differences in FMA scores were positively and significantly correlated with a correlation coefficient of r =0.431.The lesioned hemisphere was also positively correlated with the differences in FMA scores with a significant correlation coefficient of r =0.608.Conclusion PAS10 can facilitate the recovery of upper limb motor function.The change in motor cortex excitability of the contra-lesional hemisphere significantly correlates with functional recovery in the upper limb.

Effects of 20 Hz repetitive transcranial magnetic stimulation with different intensities on neurobehavior and glial fibrillary acidic protein in rats with cerebral infarction / 中华物理医学与康复杂志

Objective To investigate the effects of 20 Hz repetitive transcranial magnetic stimulation (rTMS) with different intensities on neurobehavior and expression of glial fibrillary acidic protein (GFAP) in ischemic penumbra of rats with cerebral infarction,so as to explore the probable mechanism. Methods Forty-three rats were randomly divided into a blank control group( n =7 ),a model control group( n =7),a sham stimulation control group(n =8) and a rTMS group (n =21) ; the rTMS group was further subdivided into 3 subgroups:80％ MT subgroup,100％ MT subgroup and 120％ MT subgroup,with 7 rats in each subgroup.The cerebral infarction model was established by right middle cerebral artery occlusion (MCAO) in each group except the blank control group.The 3 rTMS subgroups were given 14 successive blocks of 20 Hz rTMS with corresponding intensity.The sham stimulation control group received sham treatment (without any output).The model control group was given no stimulation,and the blank control group did not receive any special treatment.Functional assessments were performed at 3 different time points.After 14-day treatment,the expression of GFAP proteins in ischemic penumbra were detected by immunohistochemistry technique. Results Functional outcome reflected from 3 behavioral tests in 100％ MT subgroup after 14-day stimulation was better than 1 day after operation,while in the other rTMS subgroups functional outcomes were just better in 2 behavioral tests.The expressions of GFAP in 3 rTMS subgroups were all less than that in model control group. Conclusions The 20 Hz rTMS with 80％ MT and 100％ MT might be safe and effective to improve the functional outcome in rats with acute cerebral infarction,especially 100％ MT.Decrease of expression of GFAP in ischemic penumbra might be one of the mechanisms of beneficial effects of rTMS in ischemia brain injury.

Effects of high-frequency repetitive transcranial magnetic stimulation on the ultrastructure of an ischemic brain penumbra and the expression of brain-derived neurotrophic factors in rats / 中华物理医学与康复杂志

Objective To investigate the effects of high-frequency repetitive transcranial magnetic stimulation (rTMS) at different intensities on the ultrastructure of an ischemic brain penumbra and the expression of brainderived neurotrophic factor (BDNF) using rats with permanent middle cerebral artery occlusion (MCAO).Methods Forty-two rats were randomly divided into a blank control group,an MCAO model control group,a sham stimulation control group and an rTMS group.The rTMS group was divided further into 3 subgroups:an 80％ of motor threshold (MT) subgroup,a 100％ of MT subgroup and a 120％ of MT subgroup.The cerebral infarction model was established by right MCAO.rTMS treatment was given 24 hours after the MCAO model was successfully established.The rTMS group and sham stimulation control group were given 20 Hz rTMS with the planned intensities.The MCAO model control group was not given any stimulation.After 14 days of treatment,transmission electron microscopy,immunohistochemical and Western blotting ( WB ) methods were used to investigate the ultrastructure of the ischemic penumbra and the expression of BDNF.Results Damage reflected in the ultrastructure in the 3 rTMS subgroups was less than in the model control group and the sham stimulation control group.Expression of BDNF protein increased significantly in 100％ of the MT group and blank control group rats as compared with that in the sham stimulation control group,while the blank control group and the 3 rTMS subgroups had no statistically significant difference in comparison with the MCAO model control group.The expression of BDNF protein had no statistically significant difference between any of the groups.Conclusion 20 Hz rTMS might,especially at 100％ of the MT,promote the recovery of the ultrastructure of neural tissues in the ischemic penumbra after acute cerebral infarction and enhance the expression of BDNF in the ipsilesional hemisphere.This may be one of the important mechanisms of rTMS's effectiveness in the treatment of ischemic stroke.

The study on hepatocarcinoma cells cotransfected with B7.1 and SEA genes / 细胞与分子免疫学杂志

Aim To obtain the HCC cell lines which could coexpressed the B7.1 and SEA. Methods The positive clones expressing the B7.1 and SEA were screened by immunohistochemical staining. The amount of SEA in culture supernatant was detected by ELISA. Results HCC cell clones coexpressing B7.1 and SEA were obtained, and expression amount of SEA in culture supernatant reached 10～ 14× 10-8g/L. Conclusion The co-rec-ogenition immune effective system of SEA and B7.1 on HCC cells is established.

Immune responses and anti-tumor effects of melanoma antigen-n protein vaccine accompanied by CpG-containing oligodeoxynucleotide adjuvant / 中国医师杂志

Objective To prepare melanoma antigen n(MAGEn)protein vaccine and to investigate the immune responses and anti-tumor effects of MAGE-n protein vaccine accompanied by CpG-containing oligodeoxynucleotide(CpG-ODN)adjuvant.Methods The DH5? containing the MAGE-n prokaryotic expression plasmid pGEX-MAGE-n was induced and the protein was purified as protein vaccine.The CpG-ODN was synthesized as adjuvant and the C57BL/6 mice were inoculated.The cellular and humoral immune responses were detected by ELISPOT,cytotoxicity assay and enzyme linked immunosorbent assay(ELISA).The antitumor effects were detected through tumor volume and life span.Results The MAGE-n protein accompanied by CpG-ODN could induce strong MAGE-n-specific cellular and humoral immune responses.In the MAGE-n positive B16 tumor model of C57BL/6,the growth velocity of tumor was decreased and the life span was prolonged with the treatment of vaccine.Conclusion MAGE-n protein vaccine accompanied by CpG-ODN adjuvant can induce strong immune responses and anti-tumor effects against MAGE-n positive B16 tumor,which provides a new way for tumor therapy.

Internalization mediated by HSP70 receptors expressed on the surface of dendritic cells / 解放军医学杂志

Objective To study the internalization mediated by heat shock protein 70 (HSP70) receptors expressed on the surface of dendritic cells (DCs). Methods DCs from C57BL/6 mouse bone marrow were divided into HSP70 receptor-blocking group, non-receptor-blocking group and control group. The HSP70-FITC positive cells were detected by flow cytometry (FCM). Results FCM indicated that in HSP70 receptor-blocking group, the HSP70 receptors could be almost totally blocked by 50?g HSP70. With the increase of HSP70-FITC dosage, the number of HSP70-FITC positive cells increased. In non-receptor-blocking group, the rates of the HSP70-FITC positive cells were up to 90% either when 50?g HSP70-FITC or 100?g HSP70-FITC was used with no remarkable difference between them. Conclusions The internalization mediated by HSP70 receptors plays a major role in the stage of ingestion of HSP70 with the characteristics of saturation and priority. Once the receptors are blocked, non-receptor pathway participates in internalization in a dose-dependent manner.

Construction,prokaryotic expression and identification of mutant SEA(D227A) / 中国免疫学杂志

Objective:To construct the prokaryotic expression vector of SEA mutant gene SEA(D227A).The gene was expressed in E.Coli, and the induced protein was purified and identified.Methods:The SEA gene was cloned by PCR from Staphylococcus aureus strain FRI 100.D227A was introduced by changing the Asp codon GAT into GCT(Ala)in the primer.The expression plasmid pRSET SEA(D227A)was constructed and transformed into E.Coli BL21(DE3)pLysS.The induced protein was identified by Western blot.Results:The nucleotide sequence of SEA(D227A)was found to be identical to the designed sequence. E.Coli BL21(DE3)pLysS contained pRSET SEA(D227A)can express a 32 kD protein which can specially bind with the anti SEA mAb.The induced protein was purified with Ni 2+ system.Conclusion:The SEA(D227A)gene was constructed and expressed successfully.The study gave a clue to the research of low toxic superantigen. [

Induction of specific cytotoxic T lymphocytes using hepatoma antigenic peptide mixed with HSP70 in vitro / 中国免疫学杂志

Objective:To explore the possibility of inducing cell mediated immune response with HSP70 antigenic peptide complex in vitro.Methods:HSP70 peptide complex was reconstituted in vitro.Granulocyte/macrophage colony stimulating factors and interleukin 4 were used to cultivate DC from peripheral blood of HLA A2 positive healthy donors.HSP70,HSP70 peptide complex or peptide was used to activate the DC individually,which will initiate homogenize T lymphocyte to form cytotoxic T lymphocyte(CTL).The cytotxicity of the CTL was detected by MTT assay.Results:It was found that peptide specific CD8 + CTL responses were readily elicited by HSP70 peptide complex or peptide.The CTL response primed by HSP70 peptide complex was more potene than peptide alone.Conclusion:The results suggest that HSP70 peptide complex is immunogenic and HSP70 can lead to great efficient CTL response,antigenic peptides and HSP70 complex may be used as peptide vaccines for cancer immunotherapy.

Effect of HLA-I on the role of B7.1 / 中国免疫学杂志

Objective:Study the relationship of HLA I and B7 1 to elucidate the role of costimulatory molecule on antigen presentation in immunological rejection.Methods:FCM was applied to detect the expression of B7 1 Then the cytotoxicity of PBML was observed before and after blocking the HLA I Results:The level of HLA I expression increased greatly(P

Ethical issues in clinic application of tumor vaccine / 中国医学伦理学

Tumor vaccine is current hotspot of medicine. The article explicates the status quo, merit, side effects of tumor vaccine and brings up the ethic principle we should follow in clinic application.

Experimental studies on targeting-therapy of "double bomb"-Mixed monoclonal antibodies immunoconjugates against hepatocellular carcinoma / 中华消化杂志

Two highly specific monoclonal antibodies with potentul affinity against hepatocellular carcinoma HAb18, HAb25 as carriers, 131I and adriamycin (ADM) as warhead "double bomb"-mixed monoclonal antibodies immunoconjugates(131I-HAb18-ADM)/(131I-HAb25-ADM) was prepared by indirectly conjugated with dextran T-40 and chloramine-T methods. The immunoconjugating rate to target cells was 42.5%, labelling rate 49.8%, and specific radioactivity 6.88?104Bq/?g. In vitro, (131I-HAb18-ADM)/(131I-HAb25-ADM) showed stronger selective cytotoxicity against target tumor cells than either HAb18-ADM, or 131I-HAb18, or HAb25-ADM, or 131I-HAb25 conjugate alone(P

The Anti-Hepatoma Effect of Superantigen Staphylococcal Enterotoxin A Targeted by Monoclonal Antibody / 中国肿瘤生物治疗杂志

Objective: To prepare the conjugate of supcrantigen (SAg) staphylococcal enterotoxin A (SEA) and monoclonal antibody (McAb) against human hepatocellular carcinoma HAbl8 F(ab' )_(2) fragment and to investigate the anti-human hepatoma effect of peripheral blood mononuclear cells (PBMC) targeted by HAbl8 F(ab' )i-SEA. Methods: McAb HAbl8 was extracted and its F(ab' )_(2) fragment was prepared with papain; the conjugate HAblS F(ab' )_(2)-SEA was prepared with N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP); eveny collected peak after purification was identified with gel chromatography; the activity of antibody in the conjugate was identified with immunohistocheinical ABC method; the anti-hepatoma effect of PBMC targeted by HAbl8 F(ab' )_(2)-SEA was observed with MTT method. Results: The conjugate HAbl8 F(ab' )_(2)-SEA was prepared successfully and it had obvious effect of targeting PBMC to kill hepatoma cells, and this effect is correlated positively with the dose of HAbl8 F(ab')_(2)-SEA. Control groups had no such effect. Conclusion: Targeting therapy of hepatoma with McAb-SAg conjugate might be a kind of hopeful model of hepatoma im-munotherapy.

Application of activated T lymphocytes induced by hepatocellular carcinoma rejection antigenic peptide-pulsed DCs in vivo and in vitro / 中国药理学通报

AIM To study if the activated T lymphocytes induced by hepatocellular carcinoma (HCC) rejection antigenic peptide SLIVHLNEV (mutant peptide of C met) pulsed dendritic cells (DCs) can provide protection against tumor challenge in nude mice HCC model and in vitro . METHODS SLIVHLNEV was eluted from HCC cell line HHCC by mild acid buffer and was identified by the mass spectrometry technique. SLIVHLNEV pulsed DCs isolated from HLA A2+ ruptured spleen were co cutured with isogeneic T lymphocytes. Cytotoxicity was tested using lactate dehydrogenase (LDH) release assay. The induced cytotoxic T lymphocytes (CTLs) were implanted into nude mice in order to protect them against transplanted HHCC tumor cells. The inhibition effect of CTLs on the growth of implanted tumor in nude mice was also observed. RESULTS The CTLs induced by SLIVHLNEV pulsed DCs had unique ability to kill HHCC tumor cells in vitro. They also protected the nude mice against the further challenge of HHCC tumor cells and inhibited the growth of implanted tumor. CONCLUSION DC based HCC rejection antigenic peptide SLIVHLNEV vaccine can induce powerful immune reaction both in nude mice HCC model and in vitro.

The design and prediction of biology characteristics of recombinant toxin CD80-Linker-SEA / 中国免疫学杂志

Objective:To construct the eukaryotic expressed vector which express recombinant toxin CD80 Linker SEA and predict the rationality and feasibility of the linker Methods:Utilize the sequence analysis software to analyze the flexibility、antigenicity、Hoop&Woods hydrophilicity and episode of recombinant toxin CD80 Linker SEA Results:Through the analysis of the software,it could be found that the recombinant toxin has correct domains of CD80 and SEA The linker has low episode、low antigenicity and high flexibility Conclusion:The results of computer analysis could help us to rationally design the recombinant toxin CD80 Linker SEA and keep it's maximum biological activity

CORRELATION OF EBV-DNA IN CANCER CELLS AND ANTIBODY OF SERUM EBV IN PATIENTS WITH NASOPHARYNGEAL CARCINOMA IN NORTHWEST CHINA / 解放军医学杂志

Assay"'of Epstein-Barr virus (EBV) DNA was conducted in the tissues of 60 patients with nasopharyngeal carcinoma (NPC), 30 patients with chronic inflamation of nasopharyngeal mucous membrane and 40 patients with other malignant epithelial tumor respectively using DNA in situ hybridization method. The positive rate of EBV-DNA in patients of NPC was 71.6%, while it was negative in pat ents with other malignant tumors a small number of EB-VDNA positive cells were also discovered in the epithelial cells of paratumours and chronic inflamation of nasopharyngeal mucous membrane. Significant correlation was found in the study of serum EBV-antiby in patients of NPC and EBV-DNA in cancer cells. The results suggest that EBV may be of etiologic significance in the pathogenesis of NPC.