Objective To construct an eukaryotic vector with expression of human survivin gene with green fluorescent protein which is named pIRES2-EGFP/survivin and transfected into K562 cell line.Methods Using pDNR/Survivin plasmid as a template, the full length of survivin cDNA was amplified by PCR and subsequently cloned into T-A vector and then subcloned into pIRES2-EGFP vector. After identified by digestion of restrictive endonucleases, pIRES2-EGFP/survivin was further confirmed by sequencing. Then it was transfected into K562 cells with superfect reagents. The mRNA was isolated and survivin gene was detected by Western blotting. Results The exact sequences of pIRES2-EGFP/survivin vector were confirmed by digestion of restrictive endonucleases and sequencing. After transfection, the expressions of green fluorescent protein were present. The mRNA expression of survivin has been detected in transfected cells by RT-PCR. Conclusion The vector pIRES2-EGFP/survivin has been constructed and could express survivin gone in K562 cells successfully.

Objective To explore the effect of costimulatory moleculars expressed peripheral CD4+ T lymphocyts on graft-versus-host disease(GVHD) after allogeneic hematopoietic stem cell transplantation (alloHSCT). Methods 21 patients who suffered of hematology diseases or malignant solid tumors were underwent allo-HSCT and 10 normal individuals were enrolled in the study. The levels of CD28, CD80 and CD152expressions on peripheral CD4+ T lymphocytes were detected by FCM in different time(before allo-HSCT, 7 day,14 day, 21 day, 30 day after allo-HSCT, thetime of GVHD and the time after GVHD treated). STR-PCR was used to detect micro-satellites chimeras forming. Results All 21 patients achieved engraftment. By STR-PCR assay, 12 cases formed complete chimeras (CC) and 9 cases formed mixed chimeras (MC). A multivariate COX survival function modle analysis showed: CD4 CD152 was independent prognostic factors for GVHD (x2=13.128,P ＜0.0001). Patients with GVHD demonstrated higher CD4+ CD28+, CD4+ CD80+ and CD4+ CD152+ T cell levels than those without GVHD (P ＜0.01); patients with aGVHD demonstrated higher than those with cGVHD (P ＜0.05)and without GVHD (P ＜0.05); Patients with GVHD demonstrated lower CD4+ CD152+ T cell level than those without GVHD (P ＜0.01); the same result occured between aGVHD and cGVHD and without GVHD.Conclusion The incidence of GVHD between NST and traditional HSCT have no difference. B7-CD28/CD152 costimulatory pathway plays a critical role in the development of GVHD.

Objective To investigate the effect of arsenic trioxide (As2O3) and buthionine sulfoximine (BSO) on inhibiting P-glycoprotein expression in multidrug-resistant cell-K562/ADM cell. To compare the effect of As2O3 and the combined group .To determine the effect of intracellular glutathione content on the arsenic effect. Methods To investigate the effect of the arsenic group (0.5 μmol/L, 2.0 μmol/L, 5.0 (μmol/L) and/or BSO (100 μmol/L) on K562/ADM cell. Intracellular GSH contents were measured using glutathione assay kit by spectrophotometry.P-gp expression were determined by flow cytometry. Mdr-1mRNA expression were directed by semi-quantitative RT-PCR. Results P-gp expression and mdr-1mRNA expression were inhibited in 24 hours in the combination of clinic dose arsenic group (0.5 (μmol/L, 2 μmol/L)and 880(100 μmol/L). In 48 hours, the mdr-1mRNA depressive effect of the combination group (clinic dose arsenic group) was obviously stronger than high dose arsenic group. In 72 hours, the P-gp depressive effect of the combination group (clinic dose arsenic group) was obviously stronger than high dose arsenic group.Conclusion The combination of clinic dose arsenic and BSO inhibit obviously P-gp expression and mdr-1mRNA expression in K562/ADM cell.

Objective To investigate the relationship between cytokines and human graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods In 21 patients undergoing allo-HSCT,the plasma concentrations of cytokines[soluble interleukin 2 receptor(sIL-2R), interferon-gama (IFN-γ), transforming growth factor-betal (TGF-β1)] were measured by using sandwich enzyme-linked immunological assay (ELISA) and the gene expressions of three cytokines were analysed by using semi-quantitate reverse transcriptase-polymerase chain reaction(RT-PCR). Results The concentrations and gene expressions of sIL-2R and IFN-γin the patients with GVHD were significantly higher than those without GVHD (P <0.01), and they were higher in the patients with aGVHD than with cGVHD and without GVHD(P <0.05); the levels of TGF-β1 in the patients with GVHD were significantly declined(P <0.01), but in those without aGVHD were obviously increased(P <0.05). After effective treatment, unnormal sIL-2R, IFN-γand TGF-β1 expressions recovered to the levels before transplantation. A multivariate COX analysis showed sIL-2R and TGF-β1 are independent prognostic factors for GVHD (P<0.001). Conclusion Monitoring the changes of sIL-2R, IFN-γand TGF-β1 expression levels (especially sIL-2R and TGF-β1) might provide predictive markers for GVHD after allo-HSCT. The sensitivity between RT-PCR and ELISA for detecting cytokines expressions had no difference.

Objective To compare the effects of Baimai ointment and baclofen in stroke patients with spas-ticity.Methods 84 cases accompanied by limb spasticity in stroke patients by digital table were randomly divided into Baimai ointment group and baclofen group,42 cases in each group.The Baimai ointment group were treated with Baimai ointment on the spastic limbs,the baclofen group received oral baclofen tablets 30 -75mg/days for 2 weeks, 4 weeks,8 weeks.The curative effects of the two groups were compared before and after treatment.Results Before and after treatment in the two groups,the levels of spasticity,pain and activities of daily living (ADL)differences were statistically significant and Baimai ointment in the treatment of spasm.After 4 weeks and 8 weeks,the Ashworth score of the Baimai ointment group were (1.59 ±0.46)points,(0.89 ±0.56)points,and those of baclofen group were (1.75 ±0.64)points,(1.45 ±0.48)points,the differences were statistically significant(t values were 2.916, 3.367,all P <0.05).After 2 weeks,4 weeks and 8 weeks,the VAS score of the Baimai ointment group were (2.72 ± 0.54)points,(2.02 ±0.24)points,(1.24 ±0.12)points,and baclofen group were (3.56 ±0.44)points,(3.15 ± 0.48)points,(2.58 ±0.26)points,the differences were statistically significant(t values were 2.975,3.359,5.416, all P <0.05),activities of daily living (ADL)was higher than that of the baclofen group.After 8 weeks,the MBI score of the Baimai ointment group was (64.46 ±10.78)points,and baclofen group was (50.74 ±9.18)points,the difference was statistically significant between the two groups (t values was 3.562,P <0.05).Conclusion Baimai ointment has the better antispasmodic effect than baclofen in patients with stroke.

Objective:To observe any effect of dynamic motor instability training on the balance and postural control of stroke survivors.Methods:Forty stroke survivors with poor balance were randomly divided into a control group and an observation group, each of 20. In addition to routine rehabilitation, the observation group was given 20 minutes of dynamic motor instability training, 5 days a week for 8 weeks, while the control group underwent routine rehabilitation for the same length of time. Before and after the intervention, surface electromyogram of the rectus femoris, biceps femoris, and erector spinae were recorded during perturbation. Activation time and the intensity of the anticipatory and complementary postural adjustments (APAs and CPAs) were also observed. Balance and lower limb motor functioning were assessed using the Berg balance scale (BBS), the Fugl-Meyer lower extremity assessment (FMA-LE), and GaitWatch analysis.Results:After the treatment the average activation time of the rectus femoris, biceps femoris in the affected side and those of the biceps femoris [(-84.31±5.74)s] and erector spinae in the intact side in APAs were all significantly shorter in the observation group than in the control group, while the average activation intensity of the rectus femoris and erector spinae was significantly greater. There was no significant difference in the activation intensity of each muscle group in CPAs after the treatment. After the intervention the average BBS score, FMA-LE score, stride length and walking speed of the observation group all were significantly better than the control group′s averages.Conclusions:Supplementing traditional rehabilitation training with dynamic motor instability training can further improve the posture control of stroke survivors and promote recovery of their balance and walking ability.

Balance impairment is a common complication after stroke, which often leads to difficulty in walking function recovery and high risk of fall, seriously affecting the independent activity ability and quality of life of stroke patients. Accurate assessment of balance is conducive to better formulation of rehabilitation plans, evaluation of rehabilitation effects, and guidance of safer daily living activities of stroke patients. This article reviews the research progress of various methods for accurate assessment of balance function in patients with stroke.

Objective:To observe any effect of repeated unilateral high frequency transcranial magnetic stimulation (rTMS) on the swallowing function and functional magnetic resonance images of dysphagic stroke survivors.Methods:Twenty stroke survivors with dysphagia were randomly divided into a treatment group ( n=10) and a control group ( n=10). Both groups received traditional swallowing rehabilitation training, while the treatment group was additionally provided with high frequency rTMS over the cortical area of the suprahyoid muscle group of the healthy contralateral cerebral hemisphere. The stimulation frequency was 5Hz and the stimulation intensity was 80% of the resting movement threshold. During the 20-minute treatment, there was a 20-second interval between each 2-second dose of stimulation. Before and after the treatment, both groups were evaluated using the Eating Assessment Tool-10 (EAT-10), a swallowing functional communication measurement (FCM) and the modified Mann Assessment of Swallowing Ability (MMASA). In addition, functional magnetic resonance imaging was used to observe any changes in the relevant brain regions. Results:After the treatment the average EAT-10, FCM and MMASA scores of both groups were significantly better than those before the treatment, with the treatment group′s averages significantly superior to those of the control group. After the treatment, the activation range of the parietal lobe, the superior parietal lobule, the BA7 region and the BA40 area in the treatment group was significantly larger than before the treatment and larger than the control group′s ranges.Conclusions:Repeated high-frequency transcranial magnetic stimulation of the cortical area of the suprahyoid muscles in the contralateral cerebral hemisphere can improve dysphagia and promote the activation of brain areas related to swallowing after a stroke.

Objective To observe the effect of high-frequency repetitive transcranial magnetic stimulation on hemiplegic shoulder pain during convalescence of stroke.Methods A total of 40 patients with hemiplegic shoulder pain during convalescing stroke who were hospitalized in Rehabilitation Medicine Department of Jiaxing Second Hospital from January 2021 to June 2022 were selected and randomly divided into treatment group(rTMS group)and control group(sham rTMS group),with 20 cases in each group.Both groups were given anti-inflammatory and analgesic drugs and conventional rehabilitation treatment.On this basis,the treatment group was combined with rTMS treatment(stimulation frequency was 10.0Hz,stimulation intensity was 80%resting motor threshold,stimulation time was 10s,interval was 50s,total treatment time was 20 minutes,once a day from Monday to Friday,continued treatment 2 days apart,20 times as a course of treatment,a total of 4 weeks),while the control group received sham rTMS.Before treatment,2 and 4 weeks after treatment,numerical pain rating scale(NPRS)was used to assess the degree of shoulder pain,passive shoulder range of motion was measured by protractor,upper limb motor function of hemiplegic side was assessed by Fugl Meyer assessment upper extremity(FMA-UE),and the ability of daily living activities was assessed by modified Barthel index(MBI).Results After treatment,NPRS score of shoulder pain in 2 groups was significantly decreased compared with before treatment(P<0.001),passive shoulder motion and Fugl-Meyer upper limb motor function were significantly improved compared with before treatment(P<0.001),and the treatment group was significantly better than the control group,the difference between groups was statistically significant(P<0.05).The modified Barthel index in 2 groups was significantly improved compared with before treatment(P<0.001),but there was no statistical significance between 2 groups(P>0.05).Conclusion 10.0Hz high-frequency repetitive transcranial magnetic stimulation can significantly relieve hemiplegic shoulder pain during the convalession of stroke,improve the passive range of motion of the affected shoulder,and improve the upper limb motor function of the affected side,but the effect is not significant in improving the ability of daily living activities.

Objective:To evaluate the dental biofilm penetration efficiency of a novel stabilized stannous-containing sodium fluoride dentifric(EXP) and its lipopolysaccharide (LPS) neutralization efficiency.Methods:A controlled,randomized,examiner-blind in situ clinical trial was conducted with the treatment of PBS(control),EXP and a marketed stannous-containing sodium fluoride dentifrice (MKD).Fluorescent dye,fluorescent probe and fluorescence colocalization were used for sample examination and analysis.Results:EXP offered better stannous penetration into the biofilm than MKD and PBS(P ＜0.05),as well as greater LPS neutralization efficiency(P ＜0.05).There was a 96.52％ overlap of stannous ions and bounded LPS at the same sites treated by EXP.Conclusion:EXP is more effective than MKD in the delivery of stannous into the biofilm and in the neutralization of LPS.