Objective To construct an eukaryotic expression vector carrying human glutathione-S-transferase(GST)A1gene and to provide study materials for toxicology and pharmacogenetic.Methods The GST A1cDNA was amplified and separated from human liver total RNAs by RT-PCR approach and recombined with eukaryotic expression vector pcDNA3.The recombined plasmid pcDNA3-hGSTA1was verified using PCR,restriction analysis and sequencing determination.Results Human GST A1gene was recombined correctly with pcDNA3,compared with Genbank,in code152T→C,amino acid Met→Thr.Conclusion The eukaryotic expression vector pcDNA3-hGSTA1was constructed for research on toxicology and pharmacogenetics.

The effect of povidone iodine solution(PIS)on the metabolic activity of Candida albi- cans and the effect of PIS treated C.altn'cans to cultured human keratinocytes were studied by using the Alamar blue fluorescence assay.It was found that PIS inhibited the metabolic activity of Candida albi- cans with a dose-dependent manner,and that the LD_(50)of PIS for Candida albicans is 0.075%.After treating for 30 min by 2% PIS/PBS(-),Candida albicans almost lost the adherence and invasion to the cultured human keratinocytes,the wall of yeast cells became uneven and shrunken,the intracellular structures became obviously indistinct,and the yeasts could not develop to hypha form.Candida albi- cans,treated with 2% PIS in PBS(-)for 30 min or with 10% PIS Sabouraud's liquid medium for 33 hr,did not form colony when it was inoculated to Sabouraud's dextrose agar.This result suggests that the fungicidal effective of PIS is influenced by its solvent.

Objective: To investigate the surgical methods and outcomes of the enlarged translabyrinthine approach in the removal of large acoustic neuromas. Methods: A large mastoidectomy involved complete exposure of the sigmoid sinus, the dura behind the sinus for at least 1 cm, the superior petrosal sinus and the middle fossa dura. The jugular bulb was exposed and pressed downwards if necessary. The internal auditory meatus was skeletonized and uncovered for at least 270°.The debulking of the tumor began inside the anterior and inferior poles in order to find the brainstem and the facial nerve root as early as possible, and then the dissection of the nerve was done medially to laterally. Intraoperative facial nerve monitoring and postoperative CT and MRI were done in all cases. Results: Total removal was achieved in all 18 patients with tumors larger than 3 cm (mean size: 4.2 cm). There were no deaths or other complications such as intracranial infection and persistent cerebrospinal fluid leakage. There were no obvious cerebral sequelae. The facial nerve was preserved both anatomically and functionally in 14 cases, with Grade Ⅰ or Ⅱ in 8 cases, Grade Ⅲ or Ⅳ in 6 cases. Nerve interruption occurred in 4 patients who all had severe facial palsy or nerve interruption before operation. Sixteen patients resumed work within 1-3 months. Conclusion: Total removal of large acoustic neuroma could be acomplished via the translabyrinthine approach, with good preservation of facial nerve function and minimum incidence of morbidity.

BACKGROUND: Naloxone has a significant arousal effect on many types of comas. It is usually believed that this is because its inhibition on endogenous opioid peptides. But depth of coma is not necessarily positively correlated to endorphin (EP).OBJECTIVE: Based on existing findings on direct stimulating effect of naloxone on cerebral cortex, further studies need to be done to explore whether it is dose-dependent or not.DESIGN: Single-factor design based on cells.SETTING: Neurology department in a university hospital and the neurology department in a hospital of a military medical university of Chinese PLA.MATERIALS: This study was completed in the Laboratory Center of Tongji Medical College, Huazhong University of Science and Technology. Thirty healthy new born Wistar rats, regardless of their gender, aging 8 - 12 days and weighing 150 -250 g, were selected.METHODS: The experiment was performed at room temperature. The perfusion slot were placed on the microscope stage, and cells with smooth surfaces, triangle or pyramidal shapes, strong refraction and more than one neurites were selected for patch clamp experiment. Patch clamp whole-cell recording technique was used to measure the pyramidal cells of the frontal lobe immediately after separated from the Wistar rats, and to investigate the fluctuations of their membrane potential of cerebral cortex neurons and the frequencies of their spontaneous electric activities after administration of naloxone at different doses.MAIN OUTCOME MEASURES: The neural excitatory reaction rate, depolarization amplitude and increasing rate of spontaneous electric activities after administration of different doses of naloxone were selected as main outcome measurements.RESULTS: The excitatory reaction rates of cerebral cortex neurons immediately after separation to doses of naloxone(100, 50, 10, 1, 0. 1 μmol/L)were 83%, 67%, 86%, 71% and 33%; while the depolarization amplitude of them were 9. 8, 9.6, 8.4, 5.2 and 1. 3 mV respectively; and the corresponding spontaneous electric activity were increased by 587% , 375% ,291%, 125% and 69%.CONCLUSION: Naloxone can induce excitatory reactions in cerebral cortex neurons directly, and the reactions have proved to be dose-dependent.

Objective To observe the clinical features and cardiac magnetic resonance (CMR)imaging characteristics in patients with cardiac amyloidosis. Methods A total of 5 patients (4 males and 1 female) with the diagnosis of cardiac amyloidosis (3 were proven by heart transplantation, 2 by endomyocardial biopsy) were evaluated by electrocardiogram, echocardiogram, chest X-ray and CMR with delayed Gadolinium enhancement. Results Echocardiograms were abnormal in all five patients; chest X-ray showed pulmonary hemorrhage ( 3 ), cardiomegaly (5), pleural effusion (3); echocardiogram showed atrial enlargement, left ventricular wall thickening, limited ventricular wall motion, etc. CMR exhibited increased thickness of the left ventricular wall, mild to moderate depression of systolic function ( mean ejection fraction: 32.5% ± 15.0% ) and bilateral atrial enlargement with restriction of diastolic ventricular filling. In all patients, there were widespread enhancement of the thickened myocardium on delayed postcontrast studies. In 4 patients, global subendocardial delayed gadolinium enhancement was found, in papillary muscles, and interventricular septa with" zebra-like" sign in 3 patients. Left ventricular transmural delayed gadolinium enhancement was found in 1 patient. Conclusions CMR shows a characteristic pattern of global subendocardial delayed gadolinium enhancement in cardiac amyloidosis. The findings may be valuable in the diagnosis of cardiac amyloidosis.

Objective To analyze the characteristics of hypertrophic cardiomyopathy (HCM) on delayed contrast-enhanced cardiac magnetic resonance imaging (CMRI). Methods All patients underwent delayed contrast-enhanced CMRI. The left ventricle was divided into 9 segments to assess the location,extent and function of the hypertrophic segments. The t test was applied for the statistics. Results Of 154 patients, delayed enhancement of hypertrophic segment was found in 95 cases and non-delayed enhancement in 59 cases. The thickness and number of hypertrophic segment in patients with delayed enhancement were larger than those with non-delayed enhancement [ (24. 8 ± 5. 5 ) mm vs (20. 4 ±3.8) mm, t = 3.82, P < 0.05; (3.3 ± 1.9) vs (2.4 ± 1.7), t = 2. 26, P < 0.05 ], and the age was younger [ (46. 0 ± 15.2) years vs (55.0 ± 11.9) years, t = - 3. 67, P <0. 05 ]. The diffuse enhancement was found in 62 patients, and confluent enhancement in 33 patients. Confluent enhancement was found in all 14 patients after the alcohol ablation procedure. Conclusion The age, thickness and number of hypertrophic segments in patients with delayed enhancement are different from those with non-delayed enhancement.

Objective:To explore the effect of laparoscope combined with choledochoscope in the treatment of cholecystolithiasis complicated with choledocholithiasis.Methods:Totally 40 patients with cholecystolithiasis complicated with choledocholithiasis in the Second People's Hospital of Anqing from January 2016 to December 2019 were divided into double mirror combined group (20 cases) and open surgery group (20 cases) according to the operation mode.The patients in the double mirror combined group were treated by laparoscope combined with choledochoscope, and patients in the control group were treated by cholecystectomy and common bile duct exploration.The operation time, amount of bleeding, time to first flatus, postoperative complications and blood indicators were compared between the two groups.Results:The amount of bleeding[(32.50±14.82)mL], the length of hospital stay[(17.30±3.34)d], and the incidence of complication[ 5%(1/20)] in the double mirror combined group were lower than those in the open surgery group[(68.50±30.82)mL, (21.15±5.18)d, 40%(8/20)], the operation time[(162.50±56.39)min] in the double mirror combined group was longer than that in open surgery group[(102.25±21.18)min], there were statistically significant differences ( t=4.707, 2.792, 4.473, χ 2=2.692, all P<0.05). The levels of alanine aminotransferase (ALT), total bilirubin (TBIL) and albumin (ALB) in the two groups after operation were lower than those pre-operation, which in the double mirror combined group were better than those in the open surgery group[double mirror combined group (preoperation and postoperation): ALT(215.35±272.00)U/L, (44.60±44.63)U/L, TBIL(38.80±43.23)μmol/L, (16.68±11.93)μmol/L, ALB(42.65±3.25)g/L, (39.58±3.78)g/L; open surgery group(preoperation and postoperation): ALT(201.78±61.14)U/L, (61.14±48.35)U/L, TBIL(80.89±91.16)μmol/L, (40.24±53.61)μmol/L, ALB(39.37±6.81)g/L, (34.11±4.78)g/L], there were statistically significant differences ( t1=2.86, 2.72, 4.02; t2=3.54, 3.89, 4.34, t3=3.56, 4, 12, 4.01, all P<0.05). There was no statistically significant difference in white blood cell count ( P>0.05). Conclusion:The clinical efficacy of laparoscope combined with choledochoscope in the treatment of cholecystolithiasis complicated with choledocholithiasis is better than open surgery.

Objective To evaluate the efficacy of MRI for assessment of re-distribution of bone marrow mesenchymal stem cells injected intramyocardially in main organs (heart, liver, spleen and kidney) under different heart status (beating or arresting) in a porcine model. Methods Bone marrow-derived mesenchymal stem cells were obtained from the male swine and labeled with iron oxide during culture. Acute myocardial infarction was created in female swine, one week later, the survivors were randomly divided into 4 groups. Cardiopulmonary bypass was set up to arrest the heart, and then labeled cells (1×108) were intramyocardially injected into the border of the infracted myocardium in group 1 (n=6). The same volume of cells was grafted into the beating heart in group 2 (n=6). In group 3 and 4, saline was injected into either the arresting or beating myocardium. Three days later, re-distribution of stem cells and cardiac function were assessed by T2*WI and cine MRI, respectively. All animals were sacrificed for histology and real-time quantitative polymerase chain reaction (RT-PCR) of sex-determining region on Y-chromosome (SRY) investigation.The ANOVA and t test was used for statistics. Results The left ventricular end-diastolic volume (LVEDV) before transplantation for group 1-4 were: (56.8±5.3),(54.8±6.8),(57.4±4.3)and(56.8±2.8) ml, and after transplantation for group 1-4 were: (65.2±5.2),(63.2±3.7),(60.2±4.7)and(62.2±4.4) ml. The left ventricular end-systolic volume (LVESV) before transplantation for group 1-4 were: (33.5±7.6),(32.3±5.3),(33.5±3.6)and(32.7±4.6) ml,and after transplantation for group 1-4 were: (37.3±5.6),(36.3±6.9),(34.3±5.4)and(36.3±8.1) ml. The left ventricular EF values (LVEF) before transplantation for group 1-4 were: (42.3±7.2)%,(41.7±6.8)%,(41.8±8.6)% and(42.7±7.7)%,and after transplantation for group 1-4 were: (44.5±8.7)%,(43.1±7.4)%,(42.8±5.6)% and(43.3±8.4)%. The myocardial infarction area (MI) before transplantation for group 1-4 were: (6.5±2.1),(6.4±1.9),(6.5±2.5)and(6.4±2.6) cm2,and after transplantation for group 1-4 were: (6.4±2.3),(6.2±2.6),(6.3±2.5)and(6.4±2.8) cm2 . There were no statistical differences before and after transplantation in these 4 groups[P values of before and after transplantation for LVEDV, LVESV, LVEF,MI were >0.05 (F= 0.277, 0.066,0.066, 0.003); and >0.05 (F= 1.137,0.182,0.021,0.008),respectively]. The T2 value of the infracted myocardium in group 1 decreased more obviously than that in group 2[(-22.3 ± 2.2) vs (-17.0 ± 0.8) ms, t=-5.489, P＜0.01], while the T2 value of the spleen decreased more significantly in group 2 than that in group 1[(-7.7 ± 0.7) vs (-13.3 ± 1.1) ms,t=9.055, P＜0.01]. The T2 values of the liver and kidney were no significant differences in group 1 and 2 (liver, t=-0.532,P>0.05 and kidney, t=-0.113,P>0.05). The results of RT-PCR in group 1 and 2 showed significant differences in heart[(150±62) vs (72±4) U/L ,P<0.05, t=3.109], spleen[(131±1) vs (233±17) U/L, P＜0.01, t=- 13.286]and liver[(17±1) vs (9±5) U/L ,P<0.01,t= 3.492]. Pathological examination demonstrated that the transplanted stem cells were positive for Prussian blue staining, which had a good correlation with MRI results. Conclusion MRI can serve as a convenient and efficient imaging method to track the migration of stem cells with SPIO labeled in early stage and evaluate its early re-distribution in vivo. Injection of bone marrow mesenchymal stem cells in the arresting heart could favor retaining more cells in the myocardium.

Objective: To explore the short and mid-term efficacy of device closure of patent foramen ovale (PFO) for treating the patients with PFO combining cryptogenic stroke (CS) and transient ischemic attack (TIA). Methods: A total of 56 PFO patients with CS and TIA receiving device closure in our hospital from 2009-05 to 2015-12 were retrospectively studied. Transthoracic echocardiography (TTE), electrocardiogram (ECG), chest X-ray were examined at 24h, 1 month, 3 and 6 months after theoperation; telephone visit was conducted every 6 months thereafter. Results: There were 54/56 PFO patients combining CS and 2 combining TIA; 53 (94.6%)patients received PFO occluder from Starway medical technology. Aspirin was used for 6 months after the operation. The patients were followed-up for the average of (34.67±23.24) months. No body suffered from post-operative stroke and TIA; no residual shunt was observed. Conclusion: The short and mid-term efficacy of device closure has been satisfactory for treating the patients with PFO combining CS and TIA; its overall clinical value should be further investigated in large population and long-term study.

Objective To define the diagnostic criteria of cardiovascular magnetic resonance imaging in distinguishing isolated left ventricular noncompaction (LVNC) from lesser degrees of hypertrabeculation. Methods Twenty-five patients with LVNC, 39 with dilated cardiomyopathy ( DCM), 16 with aortic stenosis(AS), 15 with aortic regurgitation(AR) , 19 with hypertension (HT) and 22 normal subjects were enrolled in this study. Cardiac magnetic resonance imaging was performed to evaluate the left chamber diameter, functional parameters and noncompaction or hypertrabeculation of the left ventricle in diastole with one-way ANOVA. The left ventricle was divided into 17 segments for localizing all involved segments in this present study. Results The LVNC patients had the commonest myocardial segments involved (10±2)in all subjects. Each patient with LVNC was unexceptionally associated with apical noncompaction (17th segment) , which was seldom found in the other subjects. The lateral walls including 16th, 12th and 11th segments were the most vulnerable segments in all subjects, but nobody was found to involve the basal and mid septum including 2nd, 3rd, 8th and 9th segments. The end-diastolic NC/C (noncompaction/compaction) ratio was, on average, the greatest in patients with LVNC (3.3±0.6), compared with all other subjects(AS:1.0 ±0.3, AR:1.0 ±0.3,HT:0.8 ±0.1,healthy volunteers:0.9 ±0. 2) (F = 169. 62,P <0.05). Receiver operating characteristics analysis identified the end-diastolic NC/C ratio of>2.5 as a valuable parameter to distinguish LVNC from DCM.with values for sensitivity of 96.O%(24/25)and specificity of 94.9%(37/39),respectively.The mean number of NC/C ratio>2.5 segments in the LVNC patients was 4.0 ±2.0.while 8 of 39 patients with DCM had only one segment of NC/C ratio >2.5.Conclusions MRI is all exceUent imaging modality to diagnose LVNC and distinguish LIVNC from hypertrabeeulation.The criteria of LVNC is the NC/C ratio>2.5 in two or more than two segments of free ventricular walls associated with the left ventrieular apex involved.