To express a gene of heat shock protein 70(HSP70) in E.coli , HSP70 gene was amplified by PCR. The PCR products were cloned into pUCm T vector and were sequenced; The HSP70 gene was subcloned into vector pET 21a(+) and expressed in E.coli. SDS PAGE and Western blot were employed to identify the expression of the HSP70 gene. Results showed that a fragment about 1 96kb was amplified by PCR. Sequence analysis revealed that the sequence of HSP70 was correct. The HSP70 gene was cloned into pET 21a(+) identified by enzyme digestion and PCR. SDS PAGE and Western blot showed that a M 72 000 protein was expressed and could be recognized by anti HSP70 antibody. Therefore,HSP70 gene has been successfully expressed in E.coli .

Objective To construct a fusion gene vaccine of gastric cancer MG7-Ag mimotope and heat shock protein 70, and to investigate the humoral and cellular immune response in mice induced by the vaccine. Methods The coding sequence of MG7 mimotope was incorporated with HSP70 gene at the 3' terminus by PCR amplification. Then the PCR product was cloned into pcDNA3.1(+) vector to construct the eukaryotic expression vector. The pcDNA3.1/MG7+hsp70 was sequenced to ensure the proper encoding. C57BL/6 mice were immunized with the fusion gene vaccine, and pcDNA3.1/hsp70 and empty pcDNA3.1 were used as controls. The serum was collected at the 3rd week after each immunization. Cellular ELISA was performed to evaluate the induced humoral immunity. The splenocytes were first separated at the 9th week for the assay of antigen specific CTL lysis activity by 51Cr release. Results A fragment about 2.0 kb was obtained by PCR amplification. Sequence analysis revealed that the sequence of mimotope was connected successfully to 3' terminus of hsp70 and the fusion gene was cloned into the pcDNA3.1(+) successfully. Cellular ELISA results suggested that the serum level of MG7-Ag antibody appeared in the vaccinated mice at 9th week, while no MG7-Ag antibody was detected in the controls. The results of the specific lysis rate of splenocytes showed no statistical difference between fusion gene vaccine group and control groups. Conclusion The fusion gene vaccine was constructed successfully and the specific humoral immune response was induced by the fusion gene vaccine.

Objective To investigate the changes of brain function in moderate and high myopia patients using fractional amplitude of low frequency fluctuation (fALFF),and discuss the correlation between brain function changes and clinical data of patients with myopia.Methods Totally 21 moderate and high myopia patients (myopia group),and 21 healthy volunteers (normal control group) who were matched with myopia patients in age and gender,were selected to take rs-fMRI examination.The difference of fALFF of brain functional activity in patients with myopia and normal controls was compared,and the correlation between the changes of fALFF and clinical data of patients with myopia was analyzed,Results Compared with normal control group,the fALFF values of myopia group in the region of the left inferior frontal gyrus,putamen and right inferior frontal gyrus,putamen and insula were significantly lower (all P ＜ 0.05,AlphaSim corrected).However,in bilateral cingulate gyrus,bilateral anterior cingulate gyrus,left postcentral gyrus,left superior parietal lobule and region,fALFF values were increased (all P ＜ 0.05,AlphaSim corrected).Conclusion Patients with myopia are accompanied by abnormal neuronal activity in many brain areas,which may reflect the dysfunction of language understanding and attention control in myopic patients.

Objective To investigate the relationship between abnormal metabolism of aggrecan and joint destruction in patients with rheumatoid arthritis (RA).Methods 140 RA patients with duration less than 24 months were enrolled into this study.The study also included 100 normal controls and 95 patients with other rheumatic diseases.Three monoclonal antibodies (5D4,7D4 and BC-3) of aggrecan were used to detected aggrecan catabolic fragments in serum of RA patients and the other two groups of controls by enzyme linked immunosorbent assay (ELISA),and the correlation of aggrecan catabolic fragments with joint damage were analyzed.Sharp evaluation of hand joints in RA patients were performed at baseline and after one year follow-up.Calculating the area under the receiver operating characteristic curve (ROC) was used to evaluate the sensitivity and specificity of aggrecan catabolic fragments detected in serum of RA patients.Results Both levels of 5D4 fragment and BC-3 fragment of RA group were higher than those of normal control [5D4 of RA:(5.8±2.1) ng/μl,normal control:(2.2±1.3) ng/μl;BC-3 of RA:(11.1±3.4) ng/μl,normal control:(5.0±2.1) ng/μl,F=38.65,24.07,P＜0.001).There was no difference in 7D4 fragment among three groups (F=0.589,P=0.478).Both two fragment levels of RA patients with anti-CCP positive were greater than those patients with anti-CCP negative [5D4:(5.6±1.3) ng/μl vs (4.4±1.1) ng/μl,F=21.23,P＜0.01;BC-3:(12.2±3.9) ng/μl vs (9.3±2.8) ng/μ1,F=27.14,P＜0.01].Linear Regression showed that serum fragments detected by 5D4 and BC-3,and anti-CCP positive were risk factors for Sharp deterioration after one year follow-up.The sensitivity and specificity of combined detection of two aggrecan fragments in serum of RA patients for the prediction of joint Sharp were 56.5％ and 84.2％ respectively.Positive predictive value and negative predictive value are 74.3％ and 70.6％.respectively.Application of areas of ROC to identify the best evaluation of Sharp was 0.798.Conclusion There is positive correlation between aggrecan catabolic fragments in serum and joint Sharp evaluation of RA patients.Detection of aggrecan catabolic fragments in RA patients may predict early joint destruction.

ObjectiveTo investigate the value of double-balloon endoscopy for the diagnosis of small bowel Crohn disease(CD).MethodsData of 141 patients with suspected CD undergoing double-balloon endoscopy were reviewed.Diagnosis was made based on pathological,endoscopic findings and clinic follow-up results.Detection rates and diagnosis rates of small bowel CD were compared by double-balloon enteroscopy,CT enterography and capsule endoscopy.ResultsThe detection and diagnosis rates of small bowel CD by double-balloon endoscopy were 90.8％ ( 128/141 ) and 98.4％ ( 126/128),respectively.These two variables by CT enterography were 76.0％ (19/25) and 89.5％ (17/19),and those by capsule endoscopy were 60.0％ (15/25) and 93.3％ (14/15).ConclusionDouble balloon enteroscopy has high application value for the diagnosis of small bowel CD.For those contraindicated with endoscopy,CT enterography can be considered as a preferred auxiliary diagnostic modality.

Objective To investigate the expression changes of astrocytic syntrophin in hippocampus from human mesial temporal lobe epilepsy (MTLE). Methods From April, 2015 to July, 2016, 17 cases of hippocampus, collected from temporal lobectomy, were divided into MTLE group (n=13) and non-MTLE group (n=4) according to hematoxylin and eosin staining, glial fibrillary acidic protein and neuronal nu-clei immunohistochemical staining. Immunofluorescence double labeling and immunofluorescence histochemistry were used to observe the expression of syntrophin. Results The proliferation of astrocytes increased and neurons reduced in the hippocampus of MTLE group. Syntro-phin was found in the membrane and foot processes of astrocyte, that was enriched along perivascular astrocyte end-feet domain in non-MTLE group, but lost in MTLE group. While the whole expression of syntrophin was more in MTLE group than in non-MTLE group (t=5.421, P<0.001). Conclusion The distribution of syntrophin in hippocampus astrocytes may be related to the development of MTLE.

Objective To explore the efficacy and safety of endoscopic balloon dilatation of small bowel stricture in Crohn′s disease with single balloon enteroscopy (SBE).Methods Seven Crohn′s disease patients (four males and three females with a mean age of 37 years) were underwent 10 times dilatation.Before performance,Crohn′s disease with varying degrees of intestinal obstruction was confirmed by image and pathological examinations.There were 8 severe strictures,6 strictures in jejunum.The dilatation was performed step by step with controlled radial expansiondilator (CRE) balloon.After dilatation,the scope could reach the further small bowel was taken as a standard.Results The overall suecess rate was 8 of 10.The average diameter of intestinal stricture was expanded from (4.1 ± 2.5) mm before therapy to (10.2 ±1.5) mm (P＜0.01),and the average distance of the further small bowel that the scope reached was 62 cm.Two patients underwent twice and three times of dilatation.After the performance,all patients′ symptoms of abdominal pain and obstruction relieved obviously and there were no severe complication of bleeding or perforation.After the performance,the average follow-up time was 14 months,two patients were treated by surgery and the other two underwent dilatation treatment again because of the recurrence of symptoms.The median symptom-free time was 15 months and 70％ of patients were without symptom recurrence in one year.Conclusion Endoscopic balloon dilatation of small bowel stricture with SBE is a new technology of endoscopic treatment,which is safe and effective in the treatment of Crohn's diseasecomplicated with intestinal obstruction.

Objective To evaluate the utility of short tau inversion recovery (STIR) sequence in the diagnosis of hippocampal sclerosis ( HS).Methods Twenty-one patients with medial temporal lobe epilepsy without neoplasm lesions or injuries by conventional MRI sequence including T1WI,T2WI and FLAIR were included in this study.STIR imaging in axial,coronal and sagittal sequences was performed on these patients. Diagnosis of HS was based on the findings of hippocampal atrophy,alteration signal,disturbed internal structure and enlargement of the inferior horn. The findings shown on conventional MRI were compared with those on STIR sequence. Furthermore,the correlation of radiologic and histological findings was investigated in 6 patients operated for refractory seizures. Results On conventional MRI sequence,14 patients (66.7％) were confirmed with unilateral HS and 4 patients were suspected with unilateral HS. In contrast,all these 18 patients (85.7％) were confirmed with unilateral HS by STIR.Particularly,STIR sequence delineated the internal structure of hippocampus more clearly than conventional MRI sequences did. C shaped contour in subiculum-CA1-CA2 was revealed in normal hippocampus on STIR sequence and disappeared in HS,correlated to the pathology finding of loss of neuron in CA1 in resected tissues in 6 operated patients.The patients with HS also showed areas of hypodensity in CA4 on STIR,in accordance with pathologic findings of gliosis in this area in the 6 operated patients.Conclusion STIR sequence could depict the internal anatomical structure of hippocampus with high resolution superior to conventional MRI sequences,and can be of great value in the diagnosis of HS.

Objective In the context of worldwide shortage of amytal,explore the intracarotidpropofol test for lateralizing language area and assessing hemispheric memory function.Methods Fourteen patients with refractory partial epilepsy who were candidates for surgical intervention were included in the study.With guide under a digital subtraction angiography,propofol was injected in bilateral intracarotidsequentially.Muscle power deceasing to level 0 at the contralateral limb and eyes gazing to contralateral side were used as the mark of hemispheric anesthesia completely.The immediate language alterations were recorded.To evaluate the bilateral language and memory functions,the visual and auditory memory tasks were performed sequentially once patient could concentrate his attention ; and after limb muscle power recovering to normal level,patients were required to perform a free recall test.Any abnormal responses were recorded.Results Language dominant hemisphere was determined in 14 patients.Nine patients were confirmed as left language dominance,2 patients were right language dominance.The remained 3 patients were considered as bilateral language dominance.Meanwhile,the hemispheric memory function was able be evaluated in 13 patients.More than 67％ memory function was sustained in hemisphere contralateral to mesial temporal lesions.Transient responses including eye pain,facial muscle spasms,laughers and involuntary movements were observed.Conclusion Hemispheric language and memory functions can be assessed with direct intracarotidpropofol injection,and propofol could be an alternative drug to amobarbital used in the Wada test.

Objective:To investigate the incidence, drug sensitivity and drug resistance characteristies, and theraputic effect of staphylococcal peritoneal dialysis-associated peritonitis (PDAP), aim to provide clinical evidences for standardizing treatment therapy of staphylococcal PDAP. Methods:Clinical data of PDAP patients admitted to the Second Hospital of Jilin University, the First Hospital of Jilin University-the Eastern Division, Jilin Central Hospital and Jilin First Automobile Work General Hospital during January 1, 2013 and December 31, 2019 were retrospectively collected. The results of etiology, drug sensitivity and drug resistance of staphylococcal PDAP patients were collected. According to the pathogenic bacteria, patients were divided into staphylococcus aureus group ( n=48) and coagulase-negative staphylococcus group ( n=232). According to the results of methicillin resistance, patients were divided into drug-resistant group ( n=71) and drug-sensitive group ( n=30). The prognosis of antibiotic therapy in each group were compared. Poisson regression was used to test the changing trend of the incidence of staphylococcal PDAP. The changes of drug sensitivity and drug resistance of staphylococcus were compared between 2013 and 2019 by linear trend χ2 test. Results:A total of 1 085 cases of PDAP occurred in 625 patients were screened, and 280 cases of staphylococcal PDAP were finally included. The incidences of staphylococcal PDAP, staphylococcus aureus PDAP and coagulase-negative staphylococcal PDAP were 0.063 times per patient year, 0.010 times per patient year and 0.053 times per patient year respectively. In addition, the incidence of PDAP caused by staphylococcus, staphylococcus aureus and coagulase-negative staphylococcus decreased year by year (all P<0.05). With the change of years, the sensitivity rate of staphylococcus to rifampicin increased, while the sensitivity rate of staphylococcus to moxifloxacin decreased (both P<0.05). The drug resistance rate of staphylococcus to levofloxacin increased ( P<0.05). The staphylococcus aureus group was more prone to refractory PDAP and catheter removal than that in coagulase-negative staphylococcus group, and the recurrence rate was higher than that in coagulase-negative staphylococcus group (all P<0.05). The proportion of vancomycin used during the whole course of antibiotic therapy in drug-resistant group was higher than that in drug-sensitive group ( P<0.05). Conclusions:The incidence of staphylococcal PDAP decreases year by year, and the drug sensitivity characteristics of staphylococcus also change. The therapeutic outcomes of staphylococcus aureus PDAP are worse than that of coagulase-negative staphylococcus.