BACKGROUND: Small conductance,Ca2+-activated potassium(SK)channel,presents in various cell types and plays a crucial role in action potential profile.However,coupling and modulation of calcium and associated molecules to SK2 channel remains unclear.OBJECTIVE: To construct the recombinants of pGBAT7 and target fragments of SK2 gene,so as to observe the coupling and regulation of SK2 channel gene to calcium and other molecules.METHODS: Three pairs of primers of the target fragments of SK2 gene were designed and synthesized based on the full-length sequences of SK2.After being identified,they were individually sub-cloned into the yeast expressive plasmid pGBKT7 to construct pGBKT7-SK2 vectors.The recombinant pGBKT7-SK2 vectors were transformed into yeast AH109 by electroporation,and their activation was tested.The recombinants were extracted from yeast AH109 and verified by electrophoresis and sequencing.RESULTS AND CONCLUSION: The target fragments of SK2 gene by PCR were 411,546 and 729 bp,respectively.Three sub-clones of pGBKT7-SK2 were successfully constructed.Electrophoresis and sequencing showed that the constructed sub-clones of pGBKT7-SK2 met the expected requirements.The recombinant pGBKT7-SK2 vectors transformed into the yeast could be activated.The successful construction of the sub-clones of SK2 gene provides an important material basis for further study in the SK2 channel and function-associated molecules.

Objective:To investigate the effects of Daidzein on behavior of chronic stress depression rats and the expression of hippocampal brain-derived neurotrophic factor ( BDNF ) , neuropeptide Y ( NPY ) and non-specific immune regulation.Methods: 40 healthy adult male SD rats with body weight(210±19)g,clean grade,were chosen and fed with 1%sucrose solution for 4 d to change drinking habits.On the fifth day rats were subjected to water deprivation for 24 h without fasting.On the sixth day rats were fed with 1%surcrose solution.4 h later, preference of 1% surcrose solution was examined.According to the 1% sucrose solution preference and weight rats were randomly divided into 4 groups,normal control group(CG),model control group,(MG),fluoxetine group(FG,10.0 mg/kg),daidzein group(DG,80.0 mg/kg).At the same time of establishing model,rats were administered orally once a day for 32 d.The depression model was established by chronic unpredictable mild stress model and separation.The behavioral changes of the rats were observed, and expression of BNDF in hippocampus and NPY was measured by Western blot technology and immunohistochemistry.It was observed the proliferation function of lymphocytes,spleen index,the number of peripheral blood leukocytes and antibody-secreting cell function.Results: Compared with the normal control group(CG),the weight of rats with chronic stress protocol was lower, 1%sucrose consumption decreased,scores of rats in the open field test dropped significantly,the immobility time in the forced swimming test prolonged,the level of expression of BNDF and NPY decreased,all the differences above were statistically sig-nificant(P<0.01).Compared with model group,weight of rats in fluoxetine treatment group(FG) and daidzein treatment group(DG)in-creased,sugar consumption,scores in the open field test and the levels of expression of BNDF and NPY significantly increased,the differences were statistically significant( P<0.05 or P<0.01 ).The number of peripheral blood leukocytes and antibody-secreting cell function and proliferation of lymphocytes force in daidzein treatment group was significantly higher than the model group,daidzein dose spleen index was significantly higher than the model group(P<0.01).Conclusion: The daidzein can antagonize depressive symptoms in chronic stress mice,daidzein may increased content of BDNF in hippocampus and NPY protein, and enhanced the role of humoral immune response and lymphocyte proliferation in rats with chronic stress model.The mechanisms of antidepressant effects of daidzein might be related to the increase of content of BDNF in hippocampus and NPY protein and non -specific immune regulation.

BACKGROUND: Traditionally, cranial perforate-rinse-dram operation and tube drainage were often used in the treatment of chronic subdural hematoma in the elderly, recently,instead of which oxygen-exchange therapy through dural puncture via cranium is more and more used.OBJECTIVE: To investigate the reliability and safety of the new operation-method using oxygen-exchange in treating the older people with chronic subdural hematoma in comparison with traditional cranial perforate-rinse-dram operation.DESIGN: Retrospective case analysis.SETTING: The Department of Neurosurgery, the Second Clinical College of China Medical University.PARTICIPANTS: Eleven male patients (meanly 62 years of age)who had undertaken oxygen-exchange therapy via skull without drain tube in the Department of Neurosurgery, Second Clinical College of China Medical College from January 1997 to December 2004 were enrolled in the study, with an average disease history of 1.5 months. Among them, 10 subjects suffered from head injury to different extent within 7 weeks on average. Main chief complaint was headache, and Unilateral limb asthenia above Ⅳ was found in 5 cases asking for medical service. As shown by CT and MRI, all the subjects were diagnosed as having chronic subdural hematoma located at supratentorium, 5 cases in the right side and 6 in the left side. Volume of hematom was calculated as the following formula: volume of hematom=length×width×number of layers (1 cm thick for one layer). And the range of volume was from 70 mL to 140 mL, and the average value was 105 mL. The hematom in all the cases was found to move to the midline to different extents.METHODS: Patients in lateral recumbent position were undertaken boring at the CT-located thickest area with bone awl of 0.4-0.5 cm under local anesthesia. After boring, 14-size lumbar puncture needle with trochar was used to acupuncture dura mater then moving the needle core so that blood was discharged. Then 10 mL medical oxygen was perfused into the needle guard to cause the blood discharged from hematom again. Oxygen was perfused repetitively, once for 10 mL, till there was no blood flow. Finally, 10 mL oxygen was perfused following moving of trochar and bandaging.Oxygen volume used in each case was recorded. After operation,the volume of normal saline infusion could be increased as large as possible. The duration of infusion was 2 weeks.MAIN OUTCOME MEASURES: Improvement in limb function.RESULTS: All the patients were involved in the result analysis.①Within 24 hours, volume of hematomwas decreased obviously detected with CT, and hematom completely disappeared in 3 cases,which was replaced by oxygen. Three weeks later, all the oxygen was absorbed, the structure of midline was symmetrical and the form of brain ventricles was normal. No pain was found and 5 casesof limb asthenia were also recovered. ②Advantage and disadvantage of foramen-vertebrate oxygen-exchange operation: Advantages were listed as follows: It was simple and spent shorter time,there were few complications, and patients had no limitation in movement after operation. The operation avoided the occurrence of thrombosis of lower limbs. Cranial pressure could not lower quickly. As the pressure resident in envelope, cerebraospinal fluid could not move into the envelope. Along with the absorption of oxygen,hematom was decreased gradually till completely disappeared. Occurrence of clinical symptoms resulting from cerebral blood perfusion was decreased so as to draw rein. The disadvantage of this operation was that it was not suitable for heart disease patients to undertake this operation at bedside, and headache or limb asthenia could not be alleviated immediately.CONCLUSION: The new operation-method of cranial foramen-vertebrate oxygen-exchange to treat chronic subdural hemtoma in the elderly is safe, reliable and feasible through preoperative CT localization.

Objective To study the value of carotid plaque magnetic resonance imaging (MRI) in pre-operation assessment in the elderly patients with carotid atherosclerosis and explore the possibility of minimizing the contrast weightings to gain sweeptime. Methods Totally 70 elderly patients with cerebral ischemia (average age of 68.8 years) underwent carotid MRI and digital subtraction angiography (DSA) due to the appearance of carotid plaque detected by ultrasound. Carotid plaque MRI was acquired with 3.0T MR scanner and 8 channel surface coil. The standard carotid plague MRI program included pre-and post-contrast T1 weighted imaging (T1WI), T2 weighted imaging, proton density weighted imaging and 3D time of flight MR angiography (3D TOF MRA). All these program were divided into two combinations: the 5-sequence MRI (all the sequences) and 2-sequence MRI (T1WI and TOF MRA). Digital subtraction angiography (DSA) in coronal and lateral views of carotid artery was performed with GE Advantx LCN+. The software SPSS 13.0 was used to statistically analyze the difference between MRI and DSA, and that of two sequence combinations was used in the detection of luminal stenosis and fibrous cap (FC) rupture. Results Totally 135 arteries were analyzed while 3 arteries in one patient were excluded due to the poor quality image and stent placement. The degree of luminal stenosis were (38.3±31.0)% and (38.5±30.9)%, respectively, detected by the two MRI sequence-combination with no significant difference (t=2.447, P＞0.05) and was (35.1±31.8)% by DSA. There was a good concordance between MRI and DSA in luminal stenosis detection (Kappa value: 0.773). No statistical difference was found between two MR sequence combinations in detecting FC rupture (both in 36 vessels). DSA detected FC rupture of 16 vessels, showing remarkably difference contrast to MRI(χ2=12.0, P＜0.01). Conclusions MRI can accurately detect the luminal stenosis and FC rupture. The short time scanning resulting from sequence optimization could make MRI much more suitable than DSA to do the pre-operation assessment for senile carotid atherosclerotic patients.

Objective To study the protective effect of erythropoietin (EPO) on brain tissue with cardiac arrest-cardiopulmonary resuscitation (CA-CPR) and its mechanism.Methods 120 male Sprague-Dawley (SD) rats were randomly divided into three groups (each n =40),namely:sham group,routine chest compression group,and conventional chest compression + EPO group (EPO group).The rats in each group were subdivided into CA and 6,12,24,48 hours after restoration of spontaneous circulation (ROSC) five subgroups (each n =8).The model of CA was reproduced according to the Hendrickx classical asphyxia method followed by routine chest compression,and the rats in sham group only underwent anesthesia,tracheostomy intubation and venous-puncture without asphyxia and CPR.The rats in EPO group were given the routine chest compression + EPO 5 kU/kg (2 mL/kg) after CA.Blood sample was collected at different time points of intervention for the determination the content of serum S100 β protein by enzyme linked immunosorbent assay (ELISA).All the rats were sacrificed at the corresponding time points,and the hippocampus was harvested for the calculation of the number of S100 β protein positive cells,and to examine the pathological changes and their scores at 24 hours after ROSC by light microscopy.Results With prolongation of ROSC time,the serum levels of S100 β protein (μg/L) in the routine chose compression group and the EPO group were significantly elevated,peaking at 24 hours (compared with CA:305.7 ± 29.2 vs.44.4 ± 6.2 in routine chest compression group,and 276.7±28.9 vs.44.7±5.6 in the EPO group,both P ＜ 0.05),followed by a fall.The levels of S100β protein at each time point after ROSC in EPO group were significanthy lower than those of the routine chest compression group (83.2 ± 7.5 vs.114.3 ± 15.3 at 6 hours,123.9 ± 20.2 vs.184.9 ± 22.2 at 12 hours,276.7 ± 28.9 vs.305.7 ± 29.2 at 24 hours,256.3 ± 26.6 vs.283.2 ± 23.6 at 48 hours,all P ＜ 0.05).With the prolongation of ROSC time,the S100 β protein positive cell number in brain (cells/HP) in the routine chest compression group and the EPO group was significantly increased,peaking at 24 hours (compared with CA:14.3±2.2 vs.6.7±0.7 in the routine chest compression group,11.3± 1.3 vs.6.8±0.9 in the EPO group,both P ＜ 0.05),then it began to fall.The S100 β protein positive cell number in brain at each time point after ROSC in the EPO group was significantly lower than that of the routine chest compression group (7.0±0.9 vs.7.9± 1.9 at6 hours,8.4± 1.1 vs.10.2±2.2 at 12 hours,11.3± 1.3 vs.14.3±2.2 at24 hours,8.3±0.8 vs.10.8±2.0 at48 hours,all P ＜ 0.05).Under the light microscope,a serious brain cortex injury was found after reproduction of the model,and the degree of injury was reduced after EPO intervention.The pathological score at 24 hours after ROSC in EPO group was lower than that of routine chest compression group (3.83±0.73 vs.4.17±0.75,P ＜ 0.05).Conclusions The S100β protein level in serum and brain tissue was increased early in asphyxia CA-CPR rats.EPO intervention can reduce the expression of S100 protein and reduce the degree of brain injury.

Objective To investigate the clinical and immunological features of rapid progressive interstitial lung disease (ILD) in patients with idiopathic inflammatory myopathy (ⅡM).Methods The medical records of 146 adult ⅡM patients associated with ILD in Peking University People's Hospital from February 1996 to February 2015 were retrospectively analyzed.They were divided into three subgroups:the classic DM,PM and the clinical amyopathic dermatomyositis (CADM),and were further stratified based on with or without rapid progressive ILD (RP-ILD).Chi-squared test was used for group comparisons of categorical data and independent-sample t test for numerical data.Results ① Among 146 patients,62 (42.5％) developed RP-ILD.The prevalence of RP-ILD in CADM was signifcantly higher than dermatomyositis (DM) and polymyositis (PM).② ILD occurred after or at the same time with ⅡM in more than 90％ of patients.ILD arising before or simultaneously with DM (named early-onset ILD) tended to progress rapidly.③ For the DM group,mechanic's hands,fever,lymphopenia,hypoxemia,hypoalbuminemia,and the elevation of globulin α1 were associated with RP-ILD.CADM-ILD with weakness of swallow muscles,increasing C-reactive protein (CRP) and globulin α2,decreased vital capacity and the relatively high level of segmented granulocytes in bronchoalveolar lavage fluid tended to undergo a rapid progressive clinical course.Additionally,the decreased free triiodothyronine (FT3) and the elevation of tumor markers including carcino-embryonic antigen (CEA),neuron specific enolase (NSE),cytokerantin-19-fragment (CYFRA211) and ferritin were significantly more frequent both in DM and CADM with RP-ILD (P＜0.05) patients.④ According to multivariate analysis,mechanic's hands [HR =31.747,95％CI (2.367,425.817),P=0.009] and the elevation of CEA [HR =57.047,95％CI (1.140,2 854.885),P=0.043] and CRP [HR=31.568,95％CI (1.818,548.093),P=0.018] were potential predictive factors for RP-ILD in DM and CADM patients,respectively.Conclusion The prevalence of RP-ILD in CADM is higher than DM and PM.ILD usually occurs after or simultaneously with ⅡM,early-onset ILD in DM patients tends to progressive rapidly.Increased tumor markers and decreased FT3 indicate the deterioration of disease.Mechanic's hands and the elevation of CEA and CRP are potential predictive factors for RP-ILD in DM and CADM patients,respectively.

Epidemiology is a discipline characterized by complicated theory and practice.How to make the practice course function better is a topic worthy of exploring in educational reform for clinical students.The article explored the‘Student-Dominated’ Model based on ‘Problem-Based Learning ’ and ‘Team Based Learning ’ in teaching process and compared the model with the traditional one ( Teacher-Dominated Model) .Suggestions were given to further improve effectiveness of epidemiology practice courses.

Objective To study the clinical char acteristics and outcome of two lymphoma patients mimicking Beh?et's disease. Methods Lymphoma was diagnosed in two patients mimicking Beh?et's disease referred to our Department in 2015. A search on published similar cases in Chinese database and the Pubmed was also performed and then analyzed. Results Eight patients were indentified in this pooled analysis, six of which were non-Hodgkin lymphoma (NHL). All of the eight cases presented with cutaneous lesion, seven cases with fever, seven cases with oral ulceration and six cases with orogenital ulceration, respectively. Ocular involvement was present in four of the eight cases, two were with a positive pathergy test. Among feverish patients, six were moderate or high fever, four were high fever, one was low-grade fever. Neutropenia was found in four patients, and lymphocytoponia in four of five patients with detailed data. All patients fulfilled the 2014 International Criteria for Beh?et's Disease (ICBD) with an average score of (5.8 ±1.5), ranging from 4 to 8. Survival period ranged from one month to 36 months, with an average of 8 months. Conclusion For patients diagnosed as Beh?et's disease are finally diagnosed as lymphoma. For patients with Beh?et's disease present-ation but also present with mediate to high fever, atypical deepseated ulcer, neutropenia or lymphocytoponia, malignancy especially lymphoma should be investigated.

Objective: To investigate the relationship between human bocavirus 2 (HBoV2) infection and acute diarrhea in children younger than 5 years of age in a case-control study. Methods: This was a prospective case-control study. During May 2016 to December 2016, fecal specimens were collected from children ≤5 years of age with acute diarrhea who visited the Affiliated Children's Hospital of Capital Institute of Pediatrics (case group), or from children ≤5 years of age without diarrhea from Longtan Community Medical Service Center, Beijing (control group). The case group (n=240) and the control group (n=240) were divided into 8 age subgroups: ≤1 month old, >1-3 months old, >3-6 months old, >6-12 months old,>1-2 years old,>2-3 years old,>3-4 years old and >4-5 years old, and there were 30 cases in each age subgroup. The specimens were tested for 7 types of diarrhea-associated viruses, especially for HBoV2 by real-time PCR method. The HBoV2 viral load was predicted according to the cycle threshold (Ct). Finally, t-test was used to compare the differences between groups. Results: In the case group (n=240), the positive rate of norovirus was 16.7% (40 cases); rotavirus, 10.8% (26 cases); HBoV2, 7.5% (18 cases); adenovirus, 7.1% (17 cases); astrovirus, 6.3% (15 cases); parachovirus, 3.8% (9 cases); and Aich virus, 0.4% (1 case). The positive rates of HBoV2 in case group (7.5%, 18 cases) and control group (5.0%, 12 cases) showed no significant difference (χ²=1.280, P=0.258), as well as in different age groups (all P>0.05) . However, the mean viral load of the HBoV2 in the case group (1×10⁹copies/L with cycle threshold (Ct) 25.8) was higher than that of control group (1×10⁵copies/L with Ct 33.8), showing a significant difference (t=0.597, P=0.000). Conclusions Norovirus and rotavirus are still the important viral pathogens in children with acute diarrhea. A higher load of HBoV2 may indicate a higher risk of acute diarrhea in children ≤5 years of age in Beijing.