Objective:To generate Epstein-Barr virus(EBV) Latent Membrane Protein 2A(LMP2A) recombinant adenovirus,and provide for further investigation on the therapy vaccine against EBV associated malignancies.Methods:Full length cDNA of encoding LMP2A of EBV had been amplified by reverse transcription-PCR and cloned into pGEM-T vector.The encoding cDNA of LMP2A was inserted into E1,E3-substituted adenovirus vector pAX1CW,then the LMP2A recombinant adenovirus vector was contransfected into 293 cells togetherwith EcoT221 digested Ad5-TPC.The LMP2A recombinant adenovirus was generated by homologous recombination,and primarily identificated by ClaI enzyme digestion.The expression of LMP2A on CV1 cells infected with recombinant adenovirus analyzed by fluorescence-activated cell sorting(FACS) and confocal microscope.Results:The replication-deficient LMP2A recombinant adenovirus was generated efficiently with the titers of 2.3?10 8 pfu/ml.The LMP2A could be seen on CV1 cells membrane with confocal microscope 48 h post infected with recombinant adenovirus and the percentage of CV1 cells expressing LMP2A was 94.4% by means of FACS analysis.Conclusion:These suggested that LMP2A could be expressed efficiently by recombinant adenovirus mediated transfer,and it was the foundation of further researching in its function and developing the suitable genetic engineering vaccine against EBV associated malignancies.

Objective To investigate the effects of co-exposure to LPS and heat on plasma tumor necrosis factor-? (TNF-?) and interleukin-6 (IL-6) in rats. Methods Eighty male pathogen-free Wistar rats were randomly assigned to 4 groups: saline-injected normothermic control (group C), saline-injected heat exposed (group H), LPS-injected normothermic control (group L), LPS-injected heat exposed (group HL). Rectal temperature (Tr), heart rate (HR), mean arterial pressure (MAP), and respiratory rate (RR) were continually monitored. Plasma levels of TNF-? and IL-6 were determined at 0, 40, 80, 120 min after treatment. Results The rats in group HL displayed significantly higher values of Tr , HR , and RR and lower values of MAP than that of group C at 120 min. There was a significant difference in the values of HR and MAP between group HL and the other 3 groups at the same time point. The rats in group HL displayed an early rise in levels of plasma TNF-?, IL-6 at 40 min. The significant elevation of the peak TNF-? level at 80 min in group HL was observed. Plasma IL-6 hyperexpression was shown in rats in group HL which was significantly higher than the other 3 groups at the same time point. Conclusion Co-exposure to LPS and heat induces the rat to develop and augment systemic inflammatory response syndrome.

Objective:To prepare biotin-poloxamer(BP) conjugate micelles for epirubicin through biotin conjugated on poloxam-er. Methods:Epirubicin(EPI) was encapsulated in BP micelles. The EPI-loaded BP micelles were characterized by its particle size, zeta potential,surface morphology,as well as the efficiency of drug loading and drug encapsulation and drug release. Marrow leukemia HL-60 cells were used to evaluate the cell cytotoxicity of EPI-loaded BP micelles in vitro. The tumor model in nude mice was estab-lished through the subcutaneous injection of HL-60 cells, and then the inhibitory effect of EPI-loaded BP micelles on tumor volume growth was investigated.Results:It was found that the average particle size of EPI-loaded BP micelles was about 100 nm. In addition, the enhanced cellular uptake ability of EPI-loaded BP micelles was proved by fluorescence microscope observation. The efficiency order of the tumor volume growth inhibition was:EPI-loaded BP micelles > EPI-loaded MATP micelles> EPI-loaded poloxamer micelles>EPI. BP micelles showed significant antitumor activity and low toxicity when compared with the non-targeted micelles. Conclusion:With the advantages of EPR effect and tumor-targeting potential,BP conjugate micelles might be developed as a new system for chemo-therapeutics. However,the tumor targeting technique should be demonstrated further by the other cell experiments and large animal ex-periments.

Objective To observe the effect of surface electromyographic biofeedback (sEMG BFB) combined with routine swallow training in treating dysphagia among those with nasopharyngeal carcinoma after radiation therapy.Methods Fifty dysphagic patients with nasopharyngeal carcinoma after radiation therapy were randomly divided into a biofeedback training group and a routine treatment group,each of 25.Both groups were given routine training including orofacial function training,sensory irritation,behavioral swallowing training,and electrical stimulation.The biofeedback group was additionally given behavioral swallowing training based on sEMG BFB.Before and 4 weeks after the treatment,a videofluoroscopic swallowing study was performed to observe the opening of the upper esophageal sphincter (UES).The penetration aspiration scale (PAS) and the functional oral intake scale (FOIS) were used to evaluate the subjects' swallowing function.Results Before the treatment there were no significant differences between the two groups in terms of UES opening,average PAS score or average FOIS score.Everyone improved significantly after the treatment,but compared with the routine treatment group,UES opening was significantly better after the treatment,the average PAS score was lower and the average FOIS score was higher in the biofeedback training group.Conclusion sEMG BFB combined with routine swallowing training can improve the UES opening and swallowing ability of dysphagic patients with nasopharyngeal carcinoma after radiation therapy.

Objective To investigate the clinical efficacy and safety of erlotinib combined with concurrent whole brain radiotherapy (WBRT) in the treatment of multiple brain metastases from lung adenocarcinoma,and to provide objective evidence for improving the prognosis of patients.Methods Eighty-nine patients with brain metastases from epidermal growth factor receptor (EGFR)-mutant lung adenocareinoma who were admitted to our hospital were divided into experimental group (n =45) and control group (n=44) according to the different treatment methods.The experimental group received erlotinib combined with concurrent WBRT.The control group received oral administration of erlotinib alone for 28 d and then received concurrent WBRT.The survival rates were calculated using the Kaplan-Meier method and analyzed using the log-rank test.The other data were analyzed by the chi-square test.Results The objective response rate was significantly higher in the experimental group than in the control group (78％ vs.55％,P=0.000).The median progression-free survival (PFS) time in the experimental group and the control group were 9.1 months (95％ confidence interval [CI]:5.18-12.47) and 5.6 months (95％CI:3.46-9.12),respectively (P=0.078).The median overall survival (OS) time in the experimental group and the controlgroup were 14.3 months (95％CI:9.51-17.82) and 9.7 months (95％CI:4.59-16.74),respectively (P=0.032).The incidence rates of headache and dizziness were significantly higher in the experimental group than in the control group (38％ vs.14％,P=0.029;33％ vs.9％,P=0.020).Conclusions In the treatment of multiple brain metastases from EGFR-mutant lung adenocarcinoma,erlotinib combined with concurrent WBRT is superior to erlotinib alone.The combination therapy increases PFS and OS time of the nervous system in patients.

Objective: To investigate the effect of global end-diastolic volume index (GEDI)-guided fluid resuscitation on the prognosis of patients with chronic heart failure and septic shock. Methods: This study was a prospective randomized controlled study. Consecutive eligible patients were divided into 2 groups according to the random number table method: control group (n=21) and experimental group (n=20). On the basis of routine treatment, patients in the control group received early goal-directed therapy until the central venous pressure (CVP) reaching 8-12 mmHg (1 mmHg=0.133 kPa), mean arterial pressure reaching over 65 mmHg, urine volume reaching over 0.5 ml·kg^-1·h^-1, and central venous oxygen saturation reaching more than 70%. On the basis of routine treatment, patients in the experimental group were monitored continuously on cardiac output with pulse indication and fluid resuscitation guided by volume index GEDI. The GEDI should be maintained on the range of 680-800 ml/m². The remaining resuscitation goals were the same as control group. General clinical data of the two groups were collected at admission. Negative fluid balance onset time, duration of mechanical ventilation, ICU mortality and 28-day mortality were compared between the two groups. The outcomes were recorded as listed: start time of negative fluid balance, duration of mechanical ventilation, mortality in ICU and 28-day mortality. Results: There was no significant difference in age, sex, weight, APACHE Ⅱ score, SOFA score and NYHA functional class score between the two groups (all P>0.05). The negative liquid balance onset time in the control group was 3.5 (2.5, 4.0) days, which was significantly longer than that in the experimental group (2.6 (2.0, 3.0) days,U=115.0, P=0.012). The duration of mechanical ventilation was 355 (118, 552) hours in the control group, which was significantly longer than that in the experimental group (132 (36.75, 233.3) hours, U=130, P=0.038). The ICU mortality was 38.1% (8/21) in the control group, tended to be higher than that in the experimental group (20.0%(4/20), χ²=1.620, P=0.203). The 28-day mortality was 42.9% (9/21) in the control group, similar as in the experimental group (25.0%(5/20), χ²=1.482,P=0.477). Conclusion Fluid resuscitation guided by volume index (GEDI) may improve the prognosis of patients with chronic heart failure complicated with septic shock.

Objective To investigate the effect of 11 mHz ultra-low frequency transcranial magnetic stimulation (ILF-TMS) on neural function recovery and expressions ofnestin and Brdu in the hippocampus of cerebral ischemia and reperfusion rats.Methods Seventy-two rats were used in our study and divided into 11 mHz ILF-TMS group,sham-stimulated group and sham-operated group (n=24);The filament method was used to establish the focal cerebral ischemia and reperfusion (I-R) rat models in the first two groups.Rats in the sham-operated group only performed blood vessel separation without ligation; rats in the 11 mHz ILF-TMS group were given magnetic stimulation at 11 rnHz and 500 Gs for 15 min once daily; rats in the sham-stimulated group were only inputted magnetic stimulator.Seven,14 and 21 d after the group division,the recovery of neurological function was evaluated by 18-point nerve dysfunction scoring method,and immunohistochemistry was used to detect the nestin and Brdu positive cells in the hippocampus of rats in each group.Results The neurological deficit scores in the 11 mHz ILF-TMS group were significantly higher than those in the sham-stimulated group and sham-operated group 14 and 21 d after group after dividing (P＜0.05).As compared with sham-stimulated group and sham-operated group,1 mHz ILF-TMS group had increased number of nestin positive cells,with significant difference on the 7th,14th and 21st d; there was no statistical significance in the number of Brdu positive cells between sham-stimulated group and 1 mHz ILF-TMS group (P＜0.05).Conclusions Brain injury can stimulate the expression of nestin and Brdu in a certain degree.Although 11 mHz ultra-low frequency transcranial magnetic stimulation can improve the expression of nestin more effectively,the function of promoting cell proliferation of Brdu-positive cells is not observed.

Objective To investigate the effect of extremely low frequency-electromagnetic field (ELF-EMF) on bone mesenchymal stem cells (BMSCs) differentiating into neuron like cells in vitro and research its mechanism.Methods BMSCs were collected from rats by means of whole bone marrow adherent.Flow cytometry was used to assay cell surface marker at passage 3.And then,BMSCs were assigned into four groups:ELF-EMF group,ELF-EMF+U0126 (inhibitor) group,U0126 group and control group;cells were induced by medium (2％ DMSO and 200 μmol/L BHA) for 5 h.In the process of neural induction,ELF-EMF group and ELF-EMF+U0126 group were received 10 Hz,500 GS ELF-EMF stimulation.Besides,ELF-EMF+U0126 group and U0126 group were pretreated with 50 μmol/L extracellular signal-regulated kinase (ERK)1/2 inhibitor U0126.The morphology of BMSCs was observed under inverted microscope.The expression of nestin was detected by immunofluorescent staining and Western blotting to identify and determine the differentiation.Western blotting was applied to detect the preotein level of phosphorylase-ERK1/2 after ELF-EMF exposure.Results BMSCs presented a single long spindle morphology,growing with close whirlpoor-like arrangement;CD90 expression rate was up to 97.9％,while that of CD45 only 4.7％.After induction,each group of cells showed similar shape with neuron-like cells gradually.As compared with the other three groups,ELF-EMF group had significantly higher expression levels of nestin and phosphorylatd ERK1/2 detected by immunofluorescent staining and Westem blotting,respectively (P＜0.05).Meanwhile,the expression levels of nestin among the ELF-EMF+U0126 group,U0126 group and control group were not statistically significant (P＞0.05).Conclusion ELF-EMF could promote neuronal differentiation of mesenchymal stem cells via activation of ERK1/2 signaling pathways.

OBJECTIVETo study the prevalence of cerebral palsy (here in after referred to CP) in children aged 1 - 6 in Guangxi, China, and its epidemiologic characteristics and relevant risk factors.

METHODSInvestigations on the prevalence and etiology of CP in children at ages of 1 - 6 were conducted in Nanning, Hengxian and Qinzhou of Guangxi from June to December in 1998, with a cluster sampling.

RESULTSTotally, 150 806 children aged 1 - 6 were investigated in Nanning, Hengxian and Qinzhou of Guangxi, with 89 418 boys and 61 388 girls. Among them, 193 children were diagnosed as CP, with a prevalence rate of 1.28 per thousand, higher in boys (136 cases, 1.52 per thousand ) than in girls (57 cases, 0.93 per thousand ), with statistical significance (chi(2) = 9.536 7, P = 0.02). Logistic regression analysis showed that risk factors for CP were children who could not cry after birth (neonatal asphyxia), Apgar score less than eight, delivery at lower level of maternity hospital, number of mother's gravidity, no prenatal checks for mothers, and taking antipyretics and preserved food with salt during pregnancy.

CONCLUSIONSPrevalence of CP in children of Guangxi was at a lower level in comparison with that in other areas at home and abroad, which, maybe, was related with the lower neonatal survival and higher infant mortality.

Cerebral Palsy ; classification ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Logistic Models ; Male ; Pregnancy ; Pregnancy Trimesters ; Prevalence ; Risk Factors

Objective:To analyze the short-term therapeutic efficacy of radial extracorporeal shock wave therapy for patients with subacromial impingement syndrome.Methods:A total of 106 patients diagnosed as having subacromial impingement syndrome between October 2017 and April 2019 were randomized into a radial extracorporeal shock wave therapy (rESWT) group of 36, an exercise rehabilitation group of 35 and a conventional therapy group of 35. In addition to family exercise therapy, the rESWT group underwent 2000 to 2500 shots of extracorporeal shock wave therapy at 10 Hz and a pressure of 1.5-2.5 bar, once a week for four consecutive weeks. The exercise group was given range of motion exercises, joint control training and tendon movement training for 45 minutes, three times a week for four consecutive weeks. The conventional therapy group was treated with a laser apparatus and low-frequency electrotherapy, once a day, three times a week. Constant-Murle scores (CMSs) and the short form health survey (SF-36) were used to evaluate the clinical efficacy before and after 1 month of treatment.Results:Before the treatment there were no significant differences among the 3 groups in any of the measurements. After one month of treatment the average CMS pain score and total score of the exercise rehabilitation group were significantly better than the conventional therapy group′s averages. Moreover, the average body pain score, daily life ability, range of motion, muscle strength and total score of the rESWT group were all significantly better than the exercise and conventional therapy groups′ averages. In the SF-36 the average physical function, bodily pain, general health, and mental health scores of the rESWT groups were also significantly better than the other 2 groups′ averages.Conclusions:Radial extracorporeal shock wave therapy is superior to exercise therapy and conventional therapy for patients with subacromial impingement syndrome. It can restore shoulder joint function and improve the quality of life in one month.