Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Objective To explore the molecular mechanisms underlying therapeutic responses of the anti-proteinuria drugs from the view of podocyte molecule. Methods Adriamycin (ADR) nephropathy was induced by a single tail intravenous injection of adriamycin. Lisinopril, prednisone and all-trans retinoic acid (ATRA) were administered once a day to the adriamycin-induced nephrotic rats at the first day after adriamycin injection respectively. Renal tissue samples were collected at day 3, 7, 14, and 28 after adriamycin injection respectively. The distribution, mRNA expression and protein expression of nephrin, podocin, CD2AP and ?-actinin-4 were examined by indirect immunofluorescence, real-time PCR and Western blotting, respectively. The interactions among nephrin and podocin, nephrin and CD2AP, as well as the nephrin phosphorylation were detected by immunoprecipitation, respectively. Results Compared to the control rats, 24 h urinary protein of the ADR rats increased significantly at day 14 (P

Objective This paper is mainly to discuss accuracy and clinical application value of MDCT double-period enhanced scanning with low -tension water enteroclysis for colon cancer preoperative TNM stag-ing.Methods Sixty-two colon cancer patients with complete images and pathological data were selected in our hospital from January 2012 to May 2013 .We retrospectively analyzed CT image changes of the tumor location ,the extent of tumor invasion,the surrounding fat space,lymph node metastasis and distant metastasis.We compared them with postoperative pathology to prove the accuracy of MDCT double -period enhanced scanning with low -tension water enteroclysis.Results The results showed that its accuracy rate reached to 90.32%(56/62)in co-lon cancer preoperative Stage T,80.64%(50/62)in Stage N,and 100%(62/62)in Stage M respectively.Con-clusions MDCT double-period enhanced scanning with low -tension water enteroclysis can accurately display the site of colon cancer and determine the scope of tumor invasion ,lymph node metastasis and distant metastasis , and give more precise diagnosis of colon cancer and preoperative staging assessments .In conclusion , it can be used as the preferable method of preoperative examination in the colon cancer .

Objective To observe the clinical features, phenotypes and genotypes in a Chinese family with choroideremia (CHM). Methods A Chinese four-generation family (15 members) with CHM, including 5 patients (4 males/1 female), 2 female carriers and 8 healthy members, was enrolled in this study. Initially all family members underwent best corrected visual acuity (BCVA), indirect ophthalmoscopy, fundus fluorescein angiography, optical coherence tomography (OCT), visual field and full view electroretinogram (ERG). BCVA was followed up for 3 years. Venous blood samples were collected, and all of the 15 coding exons and flanking intron regions were amplified in the proband by polymerase chain reaction followed by direct sequencing. Protein structure was modeled based on the protein data bank and mutations in DeepView v4.0.1 to predict the effect of the mutations. A total of 180 healthy volunteers were enrolled as control group to matching CHM gene sequences. Results The visual acuity (VA) of 3/4 adult male patients began to decrease at less than 10, 10 and 30 years old, the average BCVA was 0.43. There were characteristic signs and symptoms of CHM including narrow visual field, extinguished rod and cone response in ERG, disappeared junction line and intermediate line of photoreceptor inner segment/outer segment on OCT. After 3 years, the mean BCVA decreased to 0.11. The BCVA of one young male patient was 1.0 in both eyes with minor changes fundus and visual field. The VA of the female patient began to decrease at 50 years old, her BCVA of two eyes were 0.5 and 0.25, respectively. The fundus changes were typical of CHM, with relative scotomas in the peripheral visual field of OD, and big scotomas in the OS. After 3 years, her mean BCVA decreased to 0.2. Of 2 female carriers, one had minor fundus changes (patches of pigmentary deposits, atrophy spots of retinal pigment epithelium cells), and the other was normal. A novel heterozygous c.1837G>A mutation in exon 15 of CHM was detected in the proband, which resulted in the substitution of serine by proline at codon 613 (p.D613N). Based on molecular modeling, the misfolded protein caused by the mutation might destabilize the structure of the helix that potentially could affect the global stability of the Rep-1 protein. Conclusions A novel c.1837G>A (p.D613N) mutation may be the causative mutation for CHM in this family. Female CHM carriers may have some signs and symptoms.

OBJECTIVE:To prepare buspiron hydrochloride sustained-release tablets and to study its release characterization in vitro and the factors affecting drug release.METHODS:Buspiron hydrochloride sustained-release tablets were prepared with hydroxypropyl methylcellulose(HPMC)as hydrophilic gel-matrix material and ethylcellulose(EC)as retarder by wet granulation.The impacts of releasing transmitters,contents of HPMC and EC,and viscosity on the drug release in vitro of the tablets were studied.RESULTS:For the prepared sustained release tablets,the 24h drug release amount was over 90%,and the drug release curve conformed to Higuchi equation.The more contents of HPMC and EC and the higher viscosity of HPMC in the tablets,the slower drug release velocity was obtained;but the viscosity of EC and the releasing transmitters had no significant impacts on the drug release velocity.CONCLUSION:With HPMC and EC as matrix materials,the 24h continuous drug release is available for buspirone hydrochloride sustained-tablets.

Objective To explore the molecular effect and interaction among nephrin, podocin, CD2AP and ?-actinin-4. Methods Firstly, the recombinant RNA interference (RNAi) plasmid-psiRNA-hH1GFPzeo, specifically targeting to the mRNA of nephrin, podocin, CD2AP or ?-actinin-4, was respectively tansfected into the mouse podocyte clone (MPC5) to each knockdown (KD) the expression of nephrin, podocin, CD2AP or ?-actinin-4. Molecular distributions were revealed by confocal microscopy, and the mRNA and protein expressions were detected with semi-quantitative RT-PCR and Western blotting. Results (1)In podocin KD group (siPod966 and siPod54), the mRNAs of podocin and nephrin were not detected, their protein decreased 92% and 79%, 82% and 67%, respectively. The mRNA and protein level of CD2AP increased 62% and 42%, 71% and 46%, respectively, whereas ?-actinin-4 did not change. In nephrin KD group (siNep492), the mRNA expression and protein level of nephrin were not detected, CD2AP increased 35% and 48%, respectively; and whereas podocin and ?-actinin-4 did not change. In CD2AP KD group (siCda744 and siCda21), the mRNA of expression CD2AP was not detected, and its protein level decreased 92% and 83%, the mRNA and protein of nephrin decreased 60% and 48%, 76% and 72%, respectively; podocin increased 38% and 22%, 56% and 44%, respectively; whereas ?-actinin-4 did not change. In ?-actinin-4 KD group (siAct1790 and siAct319), the mRNAs expression of ?-actinin-4 and nephrin decreased 69% and 58%, 64% and 49%, respectively; their protein level decreased 81% and 55%, 71% and 64%, respectively. However, the mRNAs of podocin and CD2AP increased 50% and 34%, 45% and 28%, respectively; and their protein level increased 64% and 46%, 65% and 42%, respectively. (2) With their expression change, the distributions of nephrin, podocin and CD2AP shifted evidently from the cell membrane surface to the nucleus circumference, whereas ?-actinin-4 showed no change, which was still localized in the cytoplasm and further extended to foot processes. Conclusion (1) Nephrin might more independently play a crucial role in the slit diaphragm complex. (2) Alpha-actinin-4 might interact direcdy or indirectly with nephrin, podocin and CD2AP. (3) The relationship among these podocyte molecules might not be spontaneous, either a single-directional or bi-directional reaction. (4) The normal localization of these podocyte molecules might depend on their normal expression quantity.

Objective To dynamically observe the expression of slit diaphragm complex molecules, including nephrin, podocin, CD2AP, and cytoskeleton protein a-actinin-4, in adriamycin-induced nephrotic (ADN) rats, and to further explore the molecular behavior of podocyte proteins during the occurrence and development of proteinuria and their possible mechanisms. Methods Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. Renal tissue samples were collected at day 3, 7, 14, and 28, respectively. The distribution, mRNA expression and protein expression of nephrin, podocin, CD2AP and a-actinin-4 were examined by indirect immunofluorescence, real-time PCR and Western blotting, respectively. Results (1) After the adriamycin injection, a significant increment of the 24-hour urinary protein was observed at day 14 and persisted up to day 28 (P

Objective To evaluate the role of α2 adrenergic receptors in dexmedetomidine-induced inhibition of lipid peroxidation during lung ischemia-reperfusion (I/R) injury in rats.Methods Thirty-two isolated rat lungs in which the model of isolated lung perfusion was successfully established,were divided into 4 groups (n=8 each) using a random number table:control group (C group),I/R group,dexmedetomidine group (D group) and dexmedetomidine plus yohimbine group (DY group).The isolated lungs were subjected to 60 min of ischemia and apnea followed by 75 min of reperfusion and ventilation to establish the model of isolated lung I/R injury.From the beginning of reperfusion,2.3 ng/ml dexmedetomidine was added to the perfusion fluid in D group,and 2.3 ng/ml dexmedetomidine and 0.4 μg/ml yohimbine (an α2 adrenergic receptor blocker) were added to the perfusion fluid in DY group.Lung specimens were obtained immediately after the end of reperfusion for determination of the wet/dry weight ratio (W/D ratio),superoxide dismutase (SOD) activity (by using modified pyrogallol autoxidation method) and malondialdehyde (MDA) content (by thiobarbituric acid method) and for examination of the pathological changes (using haematoxylin and eosin staining).Results Compared with C group,the W/D ratio and MDA content were significantly increased,and the SOD activity was decreased in I/R,D and DY groups (P＜0.05).Compared with I/R group,the W/D ratio and MDA content were significantly decreased,and the SOD activity was increased in D group (P＜0.05).Compared with DY group,the W/D ratio and MDA content were significantly decreased,and the SOD activity was increased in group D (P＜0.05).The pathological changes of lung tissues were significantly attenuated in D group as compared with I/R and DY groups.Conclusion The mechanism by which dexmedetomidine inhibits lipid peroxidation is related to activating α2 adrenergic receptors during lung I/R injury in rats.

Objective To evaluate CTA characteristics of anomalous origin of coronary artery from the pulmonary artery (ACAPA).Methods The clinical data of 24 patients with ACAPA were retrospectively analyzed,and the results of CTA were compared with operation.Results In 24 ACAPA cases,20 cases (20/24,83.33％) occurred in the left coronary artery (LCA),1 case (1/24,4.17％) was in the right coronary artery (RCA),1 case (1/24,4.17％) was in the anterior descending artery (LAD) and 2 cases (2/24,8.33 ％) were in the circumflex artery (LCX).The origins of coronary anomalies originated from the posterior wall of the pulmonary sinus or pulmonary trunk in 11 cases (11/24,45.83 ％),left wall in 7 cases (7/24,29.17％),right wall in 4 cases (4/24,16.67％),originated in the left pulmonary artery in 2 cases (2/24,8.33％).Collateral circulation:Infant type was in 5 cases,no collateral vessels between the coronary artery was observed;adult type was in 19 cases,of which 16 cases were of abnormal origin of the LCA and RCA,1 case was of LAD,2 cases were of LCX.The double LAD and coronary arteries with an intramural segment were found in 1 case respectively.Surgery were performed in 19 cases.Five cases were reviewed by CTA,1 case with anastomotic stenosis of LCX,1 case with restenosis of right ventricular outflow tract and 1 case with coronary pseudoaneurysm.Conclusion CTA can clearly show the origin of abnormal coronary artery,the distance from the ascending aorta,collateral vessels,combined with other coronary artery malformations,which can help surgical preparation of preoperative surgical approach,and postoperative follow-up.

Objective This study is to analyse the clinical feature and risk factors of morbidity after pulmonary resection for lung cancer in patients older than 70 years. Methods The clinical records of 222 patients older than 70 years who had undergone pulmonary resection for their lung cancer was reviewed. The patients were divided into 3 groups: group Ⅰ including the patients who had severe postoperative complications, group Ⅱ including the patients who had mild complications and group Ⅲ including the patients who had no complications. Moreover, the definitions were made that group A1 = group Ⅰ+ Ⅱ , group B1 = group Ⅲ, group A2 = group Ⅰ and group B2 = group Ⅱ + Ⅲ. Univariate analyses and multivariate binary logistic regressions relating postoperative morbidity to risk factors were performed between the group Al and Bl, A2 and B2, resulting in the identification of the independent risk factors for overall morbidity and major morbidity. Results Preoperative comorbidity was recorded in 161 patients (72.5%). Lobectomy (64.9% ) was the predominant surgical procedure. The median number of dissected LN was 14, with the range of 0 to 57. The overall morbidity was 63.5% , including major morbidity of 13.5%. Perioperative mortality was 1.8% (4 cases). The results of binary logistic regression analyses indicated that the independent risk factors for overall morbidity were preoperative weight loss (P =0.020), ASA score (P＜0.001), MVV (% predicted) (P=0. 020 ) and the number of dissected LN ( P = 0.004 ). The independent risk factors for major morbidity were ASA score ( P =0.003), MVV (% predicted) (P= 0.018) and the location of tumor (P=0.007). Conclusion Preoperative weight loss and numbers of dissected mediastinal lymph nodes were risk factor for lung cancer patients older than 70 years, Proper perioperative management for the elderly patients with high ASA score, low MVV (% predicted) or central tumor, could reduce the major postoperative morbidity.