温州医科大学学报
 
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2014 Vol. 44, No. 8
Published: 2014-08-25
 
 
547 Identification of hepatitis C virus-specific HLA-A*1101-and A*2402-restricted CD8+ T-cell epitopes#br
ZHONG Xiaozhi1, DUAN Zhiliang2, GUO Jianglong1, LI Qiang3, WANG Sina1, LI Jing1, LIN Rong4, WEN Jinsheng1.?
Objective: To identify hepatitis C virus (HCV)-specific HLA-A*1101- and A*2402-restricted CD8+ T-cell epitopes. Methods: HCV-specific CD8+ T-cell epitopes were predicted using T epitope prediction software (SYFPEITHI) and then HCV-specific HLA-A*1101 and A*2402-restricted epitope candidates were selected and synthesized. Immortalized HLA-A*1101-positive and A*2402-positive B cell lines were established. Based on above immortalized B cell lines, competitive peptide-binding assay was used to evaluate the binding affinity of epitope candidates for HLA-A*1101 and A*2402 molecules, respectively. Epitope candidates were used to stimulate PBMCs from HLA-A*1101-positive or A*2402-positive HCV-infected patients. Enzyme-linked immunospot (ELISPOT) and intracellular cytokine staining (ICS) assay were applied to measure the frequencies of IFN-γ-secreting cells in total PBMCs and the percentages of IFN-γ+ CD8+ T cells in total CD8+ T. Results: Of five HLA-A*1101-restricted epitope candidates, NS3_609 (ITLTHPITK) and NS2_165 (VVFSDMETK) had a high affinity for HLA-A*1101 molecules. Among three HLA-A*2402-restricted epitope candidates, NS3_373 (KCDELASKL) and NS5b_382 (YYLTRDPTI) had a high affinity for HLA-A*2402 molecules. Furthermore, high levels of NS3_609- and NS2_165-specific IFN-γ-secreting CD8+ T cells were detected in HLA-A*1101-positive HCV-infected patients, whereas high levels of NS3_373- and NS5b_382-specific IFN-γ-secreting CD8+ T cells were detected in HLA-A*2402 positive HCV-infected patients. Conclusion: Our Results strongly confirmed that NS3_609 (ITLTHPITK) and NS2_165 (VVFSDMETK) are novel HCV-specific HLA-A*1101-restricted CD8+ T-cell epitopes while NS3_373 (KCDELASKL) and NS5b_382 (YYLTRDPTI) are novel HCV-specific HLA-A*2402-restricted CD8+ T-cell epitopes.
2014 Vol. 44 (8): 547-554 [Abstract] ( 581 ) HTML (1 KB)  PDF  (0 KB)  ( 386 )
555 Changes of the expression of signaling pathway of PI3K/Akt/GSK3β and endoplasmic reticulum stress in the formation of deep tissue injury of pressure ulcer in rats
CUI Feifei1, ZHANG Jufang1, WU Haiying1, SONG Yajun1, JIANG Meiqin1, WENG Yuying1, JIANG Liping2.
Objective: To explore changes of the expression of signaling pathway of PI3K/Akt/GSK3β and expressions of ER chaperones in the deep tissue injury of pressure ulcer in rats. Methods: Fifty-four male Sprague-Dawley rats were divided into normal group and compressed groups (1 cycle, 3 cycles, 6 cycles, and 9 cycles group), natural recovery group as 1 day group, 3 days group, 5 days group and 7 days group according to the random number table, with six rats in each group. Muscle tissues underneath compression region and the same region normal group tissue were collected for analysis. Haematoxylin and eosin staining was carried out to examine the tissue histology; the expression of caspase-3 was assayed by immunohistochemical staining (Vectastain avidin-biotin complex, ABC); during different stages (experimental group and natual recovery group) the dynamic change of the ER chaperone such as GRP78, CHOP, caspase-12 and Akt, GSK3β, p-Akt, p-GSK3β in compressed muscle tissue were assayed by Western blot. Results: Under the light microscope of HE staining, the compressed muscle tissue of the experiment group showed a gradual degradation in the pathological changes, in recovery group, there were still signs of the pathlogical phenomenon; The immunohistochemical analysis indicated that the protein abundance of caspase-3 was elevated in the experiment group and natural recovery group compared with the control group, the detected immunoreactivity were generally distributed in cytoplasm. According to our immunoblot analysis, the protein expression of GRP78, CHOP, caspase-12 displayed a general ascending trend during different cycle; the protein expression of p-Akt showed first increased and then decreased; the protein expression of p-GSK3β offered first increased and then decreased with the time going, in recovery group p-GSK3β expression appeared first decreased and then increased with time going. Conclusion: The reasons of deep layer ulcers hard to heal may be related to muscle injury of the surrounding tissue; ERS and PI3K/Akt/GSK3β signaling pathway may involved in the procedure of deep ulcers.
2014 Vol. 44 (8): 555-560 [Abstract] ( 673 ) HTML (1 KB)  PDF  (0 KB)  ( 392 )
561 The experimental study on bone marrow mesenchymal stem cells transplantation in uterine scar reparation
CHEN Zhiwen, YANG Xiaojun, ZHANG Qian, ZHU Hua, HU Yan
Objective: To investigate feasibility of bone marrow mesenchymal stem cells transplantation in uterine scar reparation. Methods: Animal model of uterine scar was constructed in female SD rats. MSCs marked with Ad-GFP was injected into the site of uterine scar. Muscle tissue was obtained after two weeks of cell transplantation for HE staining. The directional differentiation situation of WMCs at the incisional part was detected with VEGF, TGF-β1 immunohistochemistry, and immunofluorescence staining for α-SMA, and uterine scar bursting pressures test. Results: Animal model of uterine scar was successfully constructed. MSCs transplantation upregulate expression of VEGF, and downregulate TGF-β1 (both P<0.01). With the aid of α-SMA immunofluorescence, transplanted MSCs demonstrated characteristic of immunotype of smooth muscle cells differentiaton. MSCs transplantation increased bursting pressures significantly (P<0.05). Conclusion: These results provide the evidence that MSCs transplantation can be used to promote regenerative reparation of uterine scar, reduce the risk of uterine scar dehiscence in the following pregnancy.
2014 Vol. 44 (8): 561-564 [Abstract] ( 597 ) HTML (1 KB)  PDF  (0 KB)  ( 462 )
565 The imaging diagnosis of isolated noncompaction of ventricular myocardium in adults
The imaging diagnosis of isolated noncompaction of ventricular myocardium in adults
Objective: To summarize the imaging features and investigate its diagnostic value to INVM in adults. Methods: Twenty cases of INVM were enrolled, 15 cases were detected by ECG, 8 cases were confirmed by CMR, and the imaging performances were retrospectively analyzed. Results: Eighteen cases of patients were of left ventricular, 2 cases were of left and right. Findings of ECG of 15 patients were shown as flows: All the patients were of left ventricular. The most impaired locations in sequence were 15 segment (n=15), 16 segment (n=15) and 17 segment (n=13). The INVM respected as prominent and excessive myocardial trabeculations and deep intratrabecular recesses. The Color Doppler flow imaging showed blood flowing into intertrabecular recesses with dim blood in them. Cardiac insufficiency and back-streaming of atrioventricular valve were found in most of patients. Findings of CMR of 18 patients were shown as flows: 6 cases were of left ventricular, 2 cases were of left and right. The most impaired locations in sequence were 12 segment (n=7), 13 segment (n=6), 14 segment (n=6), 15 segment (n=6) and 16 segment (n=6). The INVM mainly manifested as thickened myocardardium consisting of two layers: epicardial compacted thin myocardial and noncompacted thincked subendocardium, and the later expressed as prominent and excessive trabaculations and deep recesses. Enhancement of 28 segments of left ventricle was shown. Conclusion: The imaging of INVM is characteristic. CMR can clearly demonstrate the extent of abnormalities of myocardium as well as myocardial perfusion of INVM, which makes CMR better than ECG.
2014 Vol. 44 (8): 565-569 [Abstract] ( 648 ) HTML (1 KB)  PDF  (0 KB)  ( 406 )
570 Effects of sevoflurane on pulmonary inflammatory responses after cardiopulmonary bypass in early stage#br#
CHEN Feifei, MIAO Jianxia, LIN Lina
Objective: To observe the effects of sevoflurane inhalation on changing inflammatory mediators in the early stage after cardioversion in patients receiving coronary artery bypass grafting under cardiopulmonary bypass (CPB). And to provide a theoretical basis for the selection and application of general anesthesia drug. Methods: Thirty patients, ASA status II or III, 45 to 75-year- old, undergoing coronary artery bypass grafting under CPB for the first time were included and randomized into two groups: sevoflurane group (group S, n=15) and propofol group (group P, n=15). 0.05 mg/kg midazolam, 1.0μg/kg sufentanil, 0.3 mg/kg etomidate, and 0.15 mg/kg vecuronium were injected into two groups of patients and then the tracheal intubation were inserted. In group S, the sevoflurane was inhaled during the whole operation.including the CPB procedure. While in group P, propofol was administrated by Target Controlled Infusion system instead, and the other were same. Blood samples from both pulmonary artery and vein were collected immediately before CPB (T0), 5 min (T1), 15 min (T2), 30 min (T3) and 1 hour (T4) after cardioversion. The serum levels of human neutrophilelastase (HNE), IL-6 and IL-10 concentrations were measured, and the relative concentration of these mediators in pulmonary vein versus artery, which was expressed as PV/PA ratio, was calculated. Meanwhile, the duration of CPB, aorta blockage and tracheal intubation, the hospitalization in CCU, and use of the vasoactive agents were recorded. Results: No statistically significant differences were revealed in the terms of inflammatory mediators (HNE, IL-6 and IL-10) before CPB between two groups (P>0.05). After recovery of heart beating, two groups of HNE, IL-6 content in PV, PA ratio increased in varying degrees, but in S group at each time point rise extent was lower than that of P group (P<0.01 or P<0.05). IL-10 in the CPB ratio also increased significantly, and S was increased significantly than that in Pgroup, the differences were statistically significant (P<0.01 or P<0.05). As compared to group P, the frequency of using vasoactive agents were less, and the duration of tracheal intubation and hospitalization in CCU were shorter in group S (P<0.05 or P<0.01). Conclusion: The sevoflurane inhalation, possibly by attenuating inflammatory response, can effectively reduce lung injury in the early stage after coronary artery bypass grafting under CPB and shorten the hospitalization in CCU. In addition, more conducive to the stable hemodynamics during CPB
2014 Vol. 44 (8): 570- [Abstract] ( 587 ) HTML (1 KB)  PDF  (0 KB)  ( 291 )
574 The value of electrocardiographic criteria in differential diagnosis of right ventricular outflow tract ventricular arrhythmias
WANG Qiaoqiao1, LIN Jiaxuan2, LI Jin2, LI Yuechun2, LIN Jiafeng2
Objective: To evaluate the value of electrocardiographic (ECG) criteria in differential diagnosis of right ventricular outflow tract ventricular arrhythmias (RVOT VAs). Methods: The surface ECG pattern in 272 cases of right ventricular outflow tract VAs patients who underwent successful catheter ablation and compared with 72 cases of left ventricular outflow tract (LVOT) VAs patients were analyzed. Five criteria of the indexes of the R-wave amplitude<0.3 and duration<0.5, the precordial transitional zone index≥0, the V2 transition ratio<0.6 and precordial transition were observed≥V3 for diagnosing RVOT VAs and calculated their sensitivities, specificities, positive predictive values and negative predictive values. And recejver operating characteristic (ROC) curve analysis was conducted. Results: ①The corresponding sensitivities were 93.38%, 91.18%, 93.75%, 71.69% and 77.94%, respectively, the corresponding specificities were 87.50%, 88.89%, 91.67%, 88.89% and 93.06%, respectively, the corresponding positive predictive values were 96.58%, 96.88%, 97.70%, 96.06% and 97.70%, respectively, the corresponding negative predictive values were 77.78%, 72.72%, 79.52%, 45.39% and 52.76%, respectively. ②The areas under ROC curve according to different criteria were 0.904 (SE1: 0.024, 95% CI:
0.857~0.952), 0.900 (SE2: 0.024, 95% CI: 0.854~0.947), 0.927 (SE3: 0.021, 95% CI: 0.887~0.968), 0.803 (SE4: 0.027, 95% CI: 0.749~0.857), 0.855 (SE5: 0.023, 95% CI: 0.809~0.901). There were significant differences found in the comparisons between the indexes of the r-wave amplitude<0.3 and duration<0.5 and the V2 transition ratio<0.6, the precordial transitional zone index≥0 and the V2 transition ratio<0.6, the precordial transitional zone index≥0 and precordial transition≥V3 (Z statistics were 2.795, 2.685, 3.625, 2.311 respectively, all P<0.05). There were no significant differences found in the comparisons between the other criteria. Conclusion: The indexes of the r-wave amplitude<0.3 and duration<0.5 and the precordial transitional zone index≥0 are more effective than the other criterias in differential diagnosis of RVOT VAs.
2014 Vol. 44 (8): 574- [Abstract] ( 617 ) HTML (1 KB)  PDF  (0 KB)  ( 346 )
578 Correlation between EPHX2 gene polymorphisms and coronary heart disease in Chinese population#br#
HUANG Yiwei1, ZHANG Huaiqin2, LIN Bin1, WANG Jun1, MAO Yijian
 Objective: To study the correlation between EPHX2 gene polymorphisms and CHD in Chinese population. Methods: One hundred and seventy-six patients were underwent selected coronary angiography and patients having more than one major coronary vessel with at least 50% stenosis were defined as CHD. The control group consisted of 179 healthy subjects. rs2271001 polymorphisms were detected by polymerase chain reaction (PCR) and gene sequence. Results: Three kinds of genotypes at the rs2271001 were detected, and no deviation was observed from Hardy-Weinberg equilibrium. The frequency of allele G, A and genotype AG, GG and AA were increased in CHD group when compared with control group (P<0.05). Conclusion: These results suggest that rs2271001 A/G polymorphism be associated with CHD in Chinese population, the G allele may be a risk allele of CHD in Chinese Han nationality in Zhejiang province.
2014 Vol. 44 (8): 578-580 [Abstract] ( 648 ) HTML (1 KB)  PDF  (0 KB)  ( 543 )
581 Sympathetic skin response and HRV in patients with pakinson’s disease
KE Jiangqiong, SHAO Shengmin, ZHENG Yuanyuan, LI Yong, ZHANG Wenxin, WANG Xiaotong.
Objective: To explore features of sympathetic skin response (SSR) and heart rate variability (HRV) in Pakinson’sdisease patients and to observe the damage of autonomic nervous system, which provides objective evidence for early diagnosis of autonomic nerve’s electrophysiology of Parkinson’s disease. Methods: SSR and HRV were measured in 30 normal controls and 33 patients with Pakinson’s disease. Results: SSR was induced in all normal controls. SSR was not induced in 2 patients with Pakinson’s disease. Compared with normal controls, the latency and wave amplitude differences of limbs in the Pakinson disease patients had statistical significance (P<0.05). The parameters of HRV (SDNN-24 h, SDANN Index, SDNN Index, rMSSD) between the two groups had statistical significance (P<0.05). The grade of H-Y was associated with SSR. Conclusion: SSR and HRV can provide objective evidence for the early diagnosis of autonomic neuropathy in Parkinson’s disease.
2014 Vol. 44 (8): 581- [Abstract] ( 754 ) HTML (1 KB)  PDF  (0 KB)  ( 435 )
585 Application of double contrast-enhanced ultrasonography (DCUS) and 3-phase CT enhanced scanning in the diagnosis of early gastric cancer
ZHANG Yanbing1,2, LI Pihong3, LU Mingdong3, LI Liyi3, WANG Feihai3,

Objective: To explore the clinical value of spiral 3-phase CT enhanced scanning and DCUS for the diagnosis of early gastric cancer through comparison of postoperative pathology and the two examination.Methods: A retrospective analysis was performed to review the 256-slice spiral 3-phase CT enhanced scanning and double contrast-enhanced ultrasonography of 89 patients which were diagnosed as early gastric cancer by pathology in Gastrointestinal Ward of the Second Affiliated Hospital of Wenzhou Medical University during November 2010 to November 2013. The analysis results were compared with post-operative pathology to get accuracy rates, and then those rates were compared with each other. Results: The accuracy of DUCS and 256-slice spiral 3-phase CT enhanced scanning was 64.0% (57/89) vs 43.8% (39/89). The accuracy of 67 patients’ examination at No phase was 77.6% (52/67) vs 86.6% (58/67); the accuracy of 22 patients’ examination at N+ phase was 68.2% (15/22) vs 77.3% (17/22). Conclusion: DUCS is more accurate in diagnosis than 256-slice spiral 3-phase CT enhanced scanning in T staging of early gastric cancer; Though in N staging it is less accurate than 256-slice spiral 3-phase CT enhanced scanning, there is no statistical difference. Combination of the two examination is important for diagnosis of early gastric cancer.
2014 Vol. 44 (8): 585- [Abstract] ( 777 ) HTML (1 KB)  PDF  (0 KB)  ( 487 )
589 Investigation on the effect of McKenzie therapy combined with core stability training for patients with lumbar disc herniation
Zhou Yuda, Ye Xiangming, Tan Tongcai, Zhang Dawe
Objective: To investigate the effect of McKenzie therapy combined with core stability training for patients with lumbar disc herniation. Methods: Forty patients met with inclusion criteria were divided into control group and experimental group with each of 20 patients, the control group was treated with lumbar traction, physical therapy and other conventional treatment and McKenzie therapy, treatment group was treated with core stability exercise on the basis of the control group. All patients were evaluated with visual analog pain score, Oswestry Disability Index Scale, range of motion of the lumbar flexion before treatment and after treatment, the effective rate was evaluated after 6 months of treatment in both groups. Results: After treatment, the visual analog pain scores, Oswestry Disability Index and range of motion of the lumbar flexion of two groups were improved compared with pre-treatment assessment with a statistically significant (P<0.05); and there was a significant difference existed between two groups (P<0.05); Six months after treatment, the relapse rate of treatment group was significantly lower than that of the control group, and effective rate was significantly higher than that of the control group (P<0.05). Conclusion: McKenzie therapy combined with conventional rehabilitation therapy can relieve the pain and improve the functional activity in patients with lumbar disc herniation, but the core stability exercise can further improve function activity, relieve lower back symptoms and reduce relapse rate in patients with lumbar disc herniation.
2014 Vol. 44 (8): 589- [Abstract] ( 652 ) HTML (1 KB)  PDF  (0 KB)  ( 487 )
593 The clinical application of lumber plexus-paravertebral sciatic nerve block combined with intravenous anesthesia for hip fracture repair
XU Shihao1, WANG Quanguang2, CHEN Chanjuan2, XU Xuzhong2
Objective: To evaluate the clinical efficacy of lumber plexus-paravertebral sciatic nerve block combined with intravenous anesthesia as an anesthetic technique for hip fracture repair. Methods: Forty patients scheduled for hip fracture repair were randomly devided into two groups:Lumbar plexus-sciatic nerve block combined with propofol intravenous anesthesia group (B group) and intravenous general anesthesia group (G group). The B group received the lumbar plexus-sciatic nerve block combined with propofol intravenous anesthesia, and the G group received the intravenous general anesthesia. Both groups received a patient-controlled intravenous analgesia after the operaion. Visual analog scale (VAS), morphine requirements and the side effects (vomiting, pruritus and urinary retention) were recorded. Results: After hip fracture repair, the RVAS (rest visual analog scale)
and the PVAS (passive VAS) scores of 0 h, 3 h and 6 h in the B group were significant lower than that in G group (P<0.05). The morphine requirement in the first day, second day after the operaion and the fentanyl and propofol consumption during the operaion in B group were reduced significantly than that in G group (P<0.05), the difference of associated side effects (vomiting, pruritus and sore throat) between two groups was statistically significant (P<0.05). Conclusion: Lumbar plexus-paravertebral sciatic nerve block combined with propofol intravenous provides reliable anesthesia and analgesia, reduces morphine requirement and side effects after hip surgery.
2014 Vol. 44 (8): 593- [Abstract] ( 672 ) HTML (1 KB)  PDF (866 KB)  ( 694 )
596 A radiographic study of the femoral tunnel of the anatomy reconstruction of the anterior cruciate ligament
QIAN Yuenan, DONG Yilong, JIANG Gangyi, JIANG Wenhui, ZHAO Zhangwei, CAI Chunyuan, YANG Guojing.
Objective: To investigate the position of femoral tunnel anatomy reconstruction of the anterior cruciate ligament (ACL), and offer radiology data for tunnel angle selection and assessment of bone tunnel after ACL anatomy reconstruction. Methods: 30 adults’ cadaveric knee specimens were used, containing knee at least 20 cm or so, except for obvious degeneration, deformity and joint injuries, including 13 cases of males and 17 cases of females. Among them 11 cases were left side and 19 cases were right side, aging from 24 to 65 years, 37.3 years on the average. A K wire was passed in the tunnel and radiographs obtained. These radiographs were then evaluated in coronal and sagittal planes and alpha, beta (α, β) angles calculated for each of those. The α was defined as the angle between the axis of the femoral tunnel and the joint line. The β was defined as the angle between the axis of the femoral tunnel and the femoral long axis. The external aperture of femoral tunnel (OMF) was defined as the meeting point of K-wire and lateral cortex of the femur. The position of the OMF was evaluated in relation to the rectangle and the coordinate axis. The rectangle was defined using Bernard’s method and the origin of the coordinate axis was defined as lateral epicondyle of femur (LEF). The distance between the OMF and LEF in sagittal plane, defined the far-near direction as long axis of femur and the front - rear direction as perpendicular to long axis of femur. Results: The mean α in coronal plane was (49.5±4.6)°, the mean β in the sagittal plane was (31.3±3.1)°. The external aperture of femoral tunnel (OMF) was all located outside of the rectangle. The external aperture of femoral tunnel located in near lateral epicondyle of femur (3.16±2.51) mm, and front (4.25±2.16) mm. Conclusion: Anatomy reconstruction of the anterior cruciate ligament femoral tunnel outer approach do not interfere with reconstruction of the posterolateral corner. Genuflex at 120° and AMP approach to reconstruct ACL can get satisfying angle in both sagittal and coronal plane.
2014 Vol. 44 (8): 596- [Abstract] ( 638 ) HTML (1 KB)  PDF  (0 KB)  ( 483 )
600 Clinical experience of meticulous capsular dissection technique in thyroidectomy on thyroid tumor
WANG Huangen, YE Ping, FANG Sunyang, WU Zhiming.
Objective: To investigate the value of meticulous capsular dissection technique in thyroidectomy on thyroid tumor. Methods: Ninety-nine cases undergone meticulous capsular dissection technique  were enrolled in the treatment group, 96 case undergo conventional thyroidectomy were enrolled in the conventional group, two groups were compared in patients with complications. Results: The amount of bleeding, postoperative drainage volume 24 h and hoarseness in the treatment group was obviously lower than that in control group, there was statistically significant difference (P<0.05), the occurrerce rate of parathyroid injury in the treatment group patients was less than that of the control group, but the difference was not statistically significant (P>0.05). Conclusion: Meticulous membrane dissection operation is a safe and effective implementation of total thyroidectomy operation technology and worthy of promotion.
2014 Vol. 44 (8): 600-602 [Abstract] ( 633 ) HTML (1 KB)  PDF  (0 KB)  ( 355 )
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