Objective To evaluate the relationship between the mechanism of spinal monocyte chemoattractant protein-1 (MCP-1)-mediated maintenance of chronic pathological pain and synaptic transmission in spinal dorsal horns of rats.Methods Female Sprague-Dawley rats,aged 2-3 weeks after birth,weighing 150-210 g,were studied.The experiment was performed in 2 parts.Experiment Ⅰ Eighteen Sprague-Dawley rats were randomly divided into 2 groups (n =9 each) on 7 days after intrathecal catheters were inserted:phosphate buffer solution (PBS) group and MCP-1 group.PBS 10 μl was intrathecally injected in group PBS,and PBS 10 μ1 containing 100 ng MCP-1 was intrathecally injected in group MCP-1.The mechanical pain threshold was measured at 30 and 60 min before intrathecal injection,and 30,60,90,120,150 and 180 min and 1,2 and 3 days after intrathecal injection.Experiment Ⅱ The transverse spinal cord slices were prepared,and substantia gelatinosa neurons were selected for whole-cell patch-clamp recording.Electrophysiological recording was performed at 1 h of incubation with artificial cerebrospinal fluid (ACSF) and immediately after adding MCP-1:for excitatory synaptic transmission recording,MCP-1 (final concentration 100 nmol/L),N-methyl-D-aspartate (NMDA,final concentration 100 μmol/L) and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA,final concentration 20 μmol/L) were added to ACSF,and spontaneous excitatory postsynaptic currents (sEPSCs),AMPA receptors-mediated currents and NMDA receptors-mediated currents were recorded;for inhibitory synaptic transmission recording,MCP-1 (final concentration 100 nmol/L) and γ-aminobutyric acid (GABA,final concentration 1 mmol/L) were added to ACSF,and spontaneous inhibitory postsynaptic currents (sIPSCs) and GABA receptors-mediated currents were recorded.Results Compared with group PBS,the mechanical pain threshold was significantly decreased at 30 min-2 days after intrathecal injection in group MCP-1 (P＜0.01).Compared with those at 1 h of incubation with ACSF,the frequency and amplitude of sEPSCs were significantly increased,the amplitude of NMDA receptors-and AMPA receptors-mediated currents were increased,the frequency and amplitude of sIPSCs were decreased,and the amplitude of GABA receptors-mediated currents was decreased immediately after adding MCP-1 (P＜0.05).Conclusion MCP-1 enhances excitatory synaptic transmission through enhancing the function of NMDA and AMPA receptors in the posterior substantia gelatinosa neurons of the spinal cord;MCP-1 weakens inhibitory synaptic transmission through inhibiting GABA receptor function,which may be involved in MCP-l-mediated maintenance of chronic pathological pain in rats.

ObjectiveTo investigate the clinical efficacy of bidirectional Glenn shunt with noncardiopulmonary bypass(CPB) for treating congenital heart disease.MethodsThe clinical data of forty-four patients receiving bidirectional Glenn shunt from August 2005 to August 2011 were analyzedretrospectively,twenty-three cases of which were treated by bidirectional Glenn shunt with non-CPB (non-CPB group) and twenty-one with CPB (CPB group).The duration of ventilator-assisted breathing,chest drainage volume,amount of blood product usage,postoperative hospital stay and pulse oxygen saturation (SpO2) before leaving the hospital were compared and analyzed between two groups.ResultsForty-four patients were all cured.The duration of ventilator-assisted breathing,chest drainage volume and the amount of blood product usage of non-CPB group were less than those of CPB group [ (4.5 ± 1.3 ) h vs.(6.7 ± 1.5 ) h,(4.6 ± 1.4 ) ml/kg vs.(7.7 ± 3.1 ) ml/kg,( 221.8 ± 97.6 ) ml vs.( 423.4 ± 149.7 ) ml,P ＜ 0.05 ].There was no significant difference of the postoperative hospital stay and SpO2 before leaving the hospital between non-CPB group and CPB group[ (7.4 ± 2.3 ) d vs.(7.8 ± 1.9) d,0.91 ± 0.05 vs.0.88 ± 0.39,P ＞ 0.05 ].Conclusion Bidirectional Glenn shunt with non-CPB is a safe and feasible surgical approach,which is worthy of promotion.

Background Penetrating keratoplasty (PKP) has become an effective method of treatment for fungal keratitis in recent years,but the application timing of glucocorticoids after PKP is still unclear.Literature reported that the concentration of tear (1,3)-β-D-glucan in fungal keratitis was significantly higher than that in normal.Objective This study was to investigate the change of tear (1,3)-β-D-glucan before and after PKP in fungal keratitis and to explore the application duration of anti-fungal drugs and application timing of glucocorticoids.Methods This study protocol was approved by ethic committee of Affiliated Hospital of Qingdao University.A serial cases-observational study was performed from August,2011 to December,2012.Twenty eyes of 20 patients with fungal keratitis were collected in Affiliated Hospital of Qingdao University.PKP was performed in affected eyes,and the fellow health eyes served as controls.Tear of 50 μl was obtained in the controls on 1 day before operation and 1 day,7,14,21 and 28 days after operation to detect tear (1,3)-β-D-glucan levels.Results Tear (1,3)-β-D-glucan levels were (14.67±3.84)mg/L,(1 861.66±196.17) mg/L,(927.71±155.82)mg/L,(392.30±71.22)mg/L,(179.60±40.47) mg/L,(40.20± 12.46) mg/L and (15.12± 1.80) mg/L in the control group,preoperative 1 day,postoperative 1 day,7,14,21,28 days,respectively,showing a significant difference among various time points (F=883.45,P=0.00).Tear (1,3)-β3-D-glucan levels were gradually reduced with the lapse of the postoperative time,with significant differences between adjacent timepoints (t' =13.84,t =16.67,t' =11.02,t' =13.97,t' =-8.45,all at P=0.00).Tear (1,3)-β-D-glucan levels in postoperative 28 days came near that of normal control group,without significant difference between them (P =0.64).Fungal keratitis recurred in 2 eyes on the fifth and sixth day after operation,with the tear (1,3)-β-D-glucan levels of 2 350.24 mg/L and 1 992.82 mg/L,respectively.Conclusions The concentration of (1,3)-β-D-glucan in the tears increases in the eyes with fungal keratitis and drops to normal range at 28 days after PKP,indicating that the antifungal eyedrops should be applied until 4 weeks after PKP,and this is an optimal timing of using corticosteroid eyedrops to resist reject reaction.

Objective To detect the doubling time of airway smooth muscle cells of human and rat by the xCELLigence in-strument,a real time cellular analyzer.Methods The airway smooth muscle cells were separated by collagenase-pancreatin digestion from the rat airway.Then,added to the different holes of E-plate of xCELLigence instrument with the planting destiny of 3 000 cells/well.and so were the human airway smooth muscle cells.The E-plate was then placed on the xCELLi-gence instrument to monitor cell proliferation for 100 hours to calculate the doubling time by using the RTCA Software Package 2.0 software.Results The doubling time of human airway smooth muscle cell calculated by the real time cellular analyzer was 23.96±0.47 h,which was consistent with the data provided by the reference.The doubling time of rat airway smooth muscle cell was 18.62±0.15 h,and the computational process was simple,time-saving and also effective.Conclusion The xCELLigence instrument can be used to calculate doubling time of airway smooth muscle cells of human and rat, which provides experimental methods and reference data for the basic respiratory disease research.

To evaluating the imaging features and the pathological basis of low grade malignant cystic mesothelioma of the peritoneum (LGMCMP). In 14 cases of LGMCMP, radiographic imaging (X ray, CT, and MRI) was reviewed, and diagnosis was established by pathology. The majority of the patients were middle aged women. The main symptoms were abdominal pain or discomfort. CT and MRI revealed solitary cystic masses in 3 of them, multiple cystic nodules in 5, and diffuse cystic nodules in 6. It is therefore concluded that X ray, CT, and MRI are helpful in revealing the characteristics of LGMCMP and confirm the diagnosis.

Objective To compare the effects of health education on fat intake behavior and its psychology in students with different BMI,provide evidence for improving individual healthy education.Methods A experimental epidemiology was used,Twenty-nine secondary school classes were randomly selected by multi-stage cluster sampling from two cities in Zhejiang province.Overweight or obesity group included all students who were judged overweight or obesity,normal group included students who were judged normal and basic matched by age,grade,sex and stage of baseline.Intervention based on transtheoretical mode was conducted on two groups and intervention results were measured by the adolescent fat intake behavior of psychological measurement scales (Chinese version).Chi-square tests,T-tests,non-parameters tests and binary logistic regression were performed for data analyses.Results Stage of students with overweight or obesity and with normal BMI all moved forward after intervention,while the proportion of stage advance for students with overweight or obesity was further than students with normal BMI.Change strategy(2.60±0.79),pros(3.58±0.90) and self-efficacy(3.01±0.72) of students with overweight or obesity better than students with normal BMI in baseline((2.34±0.86),(3.08±0.91),(2.79±0.81)) while difference was not significant after intervention.Logistic regression analysis discovered that the promotion of change strategy moved stage forward for students with normal BMI(OR=2.168,95％ CI:1.560-3.014)and promotion of self-efficiency moved stage forward for students with overweight or obesity (OR=1.801,95％ CI:1.026-3.159).Conclusion The intensity and duration of psychological intervention is essential to be reinforced for students with normal BMI,while for students with overweight and obesity,psychological intervention should be promoted in combination with social support.

Objective To understand the infection status and gene types of Cryptosporidium among HIV/AIDS patients in Guangxi Zhuang Autonomous Region. Methods The fecal samples were collected from 285 HIV/AIDS cases in Nanning,Gui?lin,Qinzhou,Baise,Hechi cities of Guangxi and 150 HIV negative persons in Nanning City. The modified acid?fast staining and nested?PCR based on 18S rRNA were employed to detect the infection status of Cryptosporidium. The nested PCR products were sequenced,and the homology searches and identification for the gene types of Cryptosporidium were done by DNAStar soft?ware. Results The infection rate of Cryptosporidium in HIV/AIDS patients was 0.70%(2/285),and the rate of those with chronic diarrhea was 6.67%(2/30),the latter was significantly higher than that of the HIV negative persons(0,0/150)(P=0.002). Both the two HIV/AIDS patients infected with Cryptosporidium were from Guilin City. By molecular identification,the Cryptosporidium strains which the above 2 patients were infected with were Cryptosporidium andersoni and Cryptosporidium hominis respectively. Conclusions Cryptosporidium co?infection can be found in HIV/AIDS patients in Guangxi. The geno?types of the infection strains include Cryptosporidium andersoni and Cryptosporidium hominis.

Objective To explore the feasibility of using irreversible electroporation (IRE)mediating HPV16 E6 shRNA into cervical cancer cell line SiHa,and to clarify the influence of their co-effect on the proliferation of SiHa cells and its mechanism.Methods A HPV16 E6 gene specific interference sequence was inserted in pGenesil-1 to build a interference vector.10 pulses of IRE with 800 V,100 μs,and 1 Hz were applied to the suspension of SiHa cells and vectors.According to the treatment factors,control group,IRE group,pGenesil-N group,pGenesil-N+IRE group,pGenesil-E6 group and pGenesil-E6 + IRE group were set up.The expression of green fluorescent protein (GFP)and transfection efficiency were confirmed by inverted fluorescence microscope 24 h after the vector was transfected by IRE,and the expression efficancy of GFP was calculated.The expression levels of E6 mRNA and protein were detected by RT-PCR and Western blotting method which was also applied to detect the expressions of P53 and PCNA.The proliferative activity of SiHa cells was determined by CCK-8 assay.Results Enzyme digestion and DNA sequencing verified that the vectors were correctly constructed.GFP was seen under inverted fluorescence microscope 24 h after IRE transfection.Compared with IRE group,the expression levels of E6 mRNA and protein were decreased detected by RT-PCR and Western blotting method after the vectors were treated with IRE,the P53 protein expression level was increased (P < 0.05),and the PCNA expression level was decreased (P <0.05).The CCK-8 assay results showed the proliferative activity of SiHa cells in pGenesil-E6+IRE group was decreased more obviously than that in pGenesil E6 group (P <0.05).Conclusion IRE can play the role of gene transfection of mediating HPV16 E6 shRNA into SiHa cells, and their co-effect can significantly inhibit the proliferation of SiHa cells.

Objective To summarize the CT features of pulmonary mucormycosis in post hematopoietic stem cell transplantation (HSCT) patients with leukemia,to provide timely and accurate guidance for clinical treatment.Methods 9 pulmonary mucormycosis in post HSCT patients with leukemia confirmed by surgery,biopsy and sputum culture were analyzed retrospectively.Distribution,morphology and CT features of the disease were analyzed and summarized.All patients were underwent non-enhanced MSCT.Results Reversed halo sign (n=7);patchy ground glass opacity (GGO) (n =5);bilateral multiple pulmonary nodules and nodular GGO (n=3);bilateral multiple pulmonary micro-cysts with spiculate boundary (n=1);pleural effusion (n=2);pneumo-mediastinum (n 1) were seen.Two or more than two CT signs were seen in some patients.The interval time between the appeared reversed halo sign and the transplantation were 0.5-19 months in 7 patients,which median time was 10 months and 6 cases (85.7％) appeared within 18months.8 patients (88.9 ％) presented cough,expectorated white or yellow phlegm,among them,4 cases (50 ％) presented bloodstained sputum or hemoptysis.3 cases (37.5 ％) presented left chest pain.1 case was asymptomatic.6 patients (66.7 ％) presented significant increased body temperature with the range from 38.4 ℃ to 39.6 C,the median was 39 C.3 patients (33.3％) showed normal temperature.The WBC counting was between 0.16-3.31 × 109/L,the median was 0.7 × 109/L.The neutrophil cell counting was between 0-2.46 × 109/L,with the median of 0.09 × 109/L.Among them,7 patients (77.8％) found reversed halo sign with neutrophil counting between 0-1.63 × 109/L,and the median was 0.09 × 109/L.Conclusion Various imaging manifestations of pulmonary mucormycosis are seen in post HSCT patients with leukemia.When patients is in agranulocytosis phase after transplantation HSCT,it is highly suggested pulmonary mucormycosis infection when the characteristic reversed halo sign is found.

Objective To explore the characteristic of ACE gene I/D polymorphism in Han broad jumpers .Methods Distribu‐tion of ACE gene I/D polymorphism for broad jumpers was analyzed by PCR ,DNA sequencing ,Hardy‐Weinberg and SPSS ,and the results were compared with that of normal people .Results There were II ,DD and ID in gene ACE 16 ,and the Hard‐Weinberg re‐sults showed that the subjects were group representative .The ACE DD genotype and D allele in broad jumpers was statistically higher than normal controls (P<0 .05) .Conclusion The results suggest that ACE DD genotype and D allele are associated with training sensitivity of broad jumpers and could be the molecular marker for athletes choosing genetically .