Membrane proteins are the main undertakers of biofilm function, and also the most important target group for innovative drug discovery and research. About 60% of drugs targets are membrane proteins. Due to the obvious aggregation and denaturation tendency of membrane proteins in aqueous solution, it is difficult to simulate the membrane like environment to maintain the correct conformation of membrane proteins in vitro, which results in the slower-growing research on the structure and function of membrane proteins and related ligand drugs than that of water-soluble proteins. Membrane protein stabilization technology is the premise of establishing high specificity, high sensitivity and high throughput drug screening methods for membrane protein ligands, which is of great significance. In this paper, some techniques for stable separation and purification of membrane proteins are reviewed, including detergents, artificial membranes, polymers, lentiviral particles and so on, as well as their specific applications in drug screening.

OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits

Objective To preliminarily investigate the protective effect of chronic clonorchis sinesis(Cs) infestation against sepsis in Sprague Dawley(SD) rats in order to explore its underlying mechanism.Methods Chronic Cs infestation model of SD rats was reproduced by intra-gastric administration with Cs ova.Twenty rats were randomly(random number) divided into normal group(n=10) and Cs group(n=10).The proportion of differentiation in M1 and M2 macrophages were detected by flow cytometry.The expressions of Arg-1(arginine-1),FIZZ 1,iNOs and TNF-αmRNA were examined by reverse transcriptase polymerase chain reaction(RT-PCR).The cecal ligation and puncture(CLP) procedure was performed to reproduce sepsis model of SD rats.Sixty rats were randomly(random number) divided into control group,SHAM group,CLP group,Mφ+CLP group,Cs-Mφ+CLP group,and Cs-CLP group.The cumulative mortalities were calculated.The pathological changes of the lung tissue in different groups were demonstrated by HE staining.The serum levels of cytokines TNF-α and IL-10 were detected by ELISA at 0,24,48 and 72 h after CLP procedure.Results Compared with M1 peritoneal macrophages differentiation in control group(91.9%),rat peritoneal macrophages were activated to M2 differentiation(95.1%) in chronic Cs infection group.RT-PCR assay showed expression of Arg-1 and FIZZ 1 mRNA were higher in M2 macrophages,and on the contrary, the expression of iNOS mRNA expression was higher in M1 macrophages.The expression of TNF-α mRNA in M1 was significantly higher than that in M2, whereas the expression of IL-10 mRNA in M2 was higher than that in M1.The cumulative mortality of septic rats 72 h after CLP procedure were much lower in both chronic Cs infestation group and M2 macrophages adoptive transfer group(CLP group 70%vs.Mφ+CLP group 50%vs.Cs-Mφ+CLP group 30%vs.Cs-CLP group 0%,P<0.05).In these two groups,the pathological damages in lung tissues were significantly improved.The serum level of TNF-α was decreased and the anti-inflammatory IL-10 level was increased significantly in these two groups with Cs compared with other groups.Conclusion M2 macrophages polarization induced by chronic Cs infestation with M2 phenotype gene and expression of anti-inflammatory cytokine gene play key role in increasing anti-inflammatory cytokines and decreasing pro-inflammatory cytokines to allerviate organ damage and ameliorating the survival rate in septic rats.

ObjectiveTo investigate the clinical characteristics of the inpatients suffering from dengue fever in order to provide references for better diagnosis and treatment.Methods The clinical data of 158 dengue fever patients admitted to the First Affiliated Hospital of Sun Yat-Sen University from July 23rd to October 31st, 2014 during the 2014 epidemic in Guangzhou area were retrospectively analyzed, including general clinical manifestations, conventional examinations, pathogenesis, and prognosis.Results The mean age of the 158 patients was (56±20) years, with half of them over 60 years old (79 cases). Among them, 94 (59.49%) were male.① The common manifestations included fever (100%), headache (70.89%), myalgia/bone soreness (62.03%), and skin rash (54.43%). Bleeding and plasma leakage were found in 25.95% and 14.56% of the patients respectively.② Laboratory examination:leucopenia (75.32%) and thrombocytopenia (77.85%) were found, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were elevated in 57.59% and 77.85% of the patients respectively. However, elevation of blood hematocrit was rare (1.27%).③ It was found that in the acute phase (0 - 5 days of the onset), serum dengue virus antibody IgM (DF-IgM) was positive in 63.54% of the patients (61/96), and 92.62% (113/122) of patients were dengue virus RNA (DENA-RNA) positive.④ The rate of comorbidity in this study was 55.06% (87/158), including hypertension (27.22%) and type 2 diabetes (15.82%), which were the two most common co-morbidities.⑤ All the patients were given supportive therapy to prevent complications. They were also isolated for more than 5 days after onset, and at least for 24 hours after subsidence of fever in addition.⑥ The criteria for the diagnosis of severe dengue were fulfilled in 18 patients (11.39%). One patient died of massive hemorrhage from gastro-intestinal tract, and 1 patient voluntarily left hospital with untreated multiple organ dysfunction syndrome (MODS). Another 2 patients of dengue fever died from primary cardio-cerebrovascular disease, and the remaining 154 patients (97.47%) fully recovered with supportive therapy and complication prevention measures.Conclusions The clinical manifestations of inpatients with dengue fever in this study were typical, and they manifested a higher incidence of severe illness. DENA-RNA could be a sensitive indicator for early pathogenic diagnosis. With symptomatic and supportive therapy, most patients had a good outcome. However, early diagnosis and clinical interventions of severe dengue still need further studies.

Objective Observe the response and adverse effects of impact dose recombination human erythropoietin(rhEPO)in the breast cancer patients with anemia induced by chemotherapy.Methods 43 breast cancer patients with anemia induced by chemotherapy were randomly divided into two groups,treatment group(23 patients)and the control(20 patients).The treatment group received rhEPO for 10 days and oral iron therapy,while the control group was oral iron therapy only.Results After 4 weeks,the hemoglobin level, hematocrit,reticulocyte and quality of life be revaluated,the treatment group was significantly better than the control.The adverse effects was tolerable.Conclusion The impact dose rhEPO is effective and tolerable in the breast cancer patients with anemia induced by chemotherapy.

Objective To evaluate the diagnostic performance of a point-of-care testing for sensitive cardiac troponin Ⅰ (POCT-cTnI) in early diagnosis of chest pain patients who had a high pretest probability of acute myocardial infarction (AMI).Methods Total of 127 patients with new-onset chest pain at the emergency department were enrolled.Blood samples were drawn for the routine blood test,and determined POCT-cTnI and central laboratory testing for high sensitive cardiac troponin T (CLT-hscTnT) at admission,three and then at six hours after admission.All patients were divided into AMI group and non-AMI group according to the final diagnosis,which was adjudicated independently by two physicians who reviewed all available medical records for the 90-day follow-up period,and they were unaware of the results of the investigational assays of cardiac troponins.The receiver operating characteristic (ROC) curves were constructed to assess and compare the diagnostic performance of AMI of two cardiac troponin assays.The comparison of areas under the ROC curves (AUC) was performed by DeLong test,and the sensitivity,specificity,negative predictive values (NPV) and positive predictive values (PPV) for the target markers were calculated by applying a maker-specific cutoff value.Results The final diagnosis of AMI was made in 40 of 127 patients (31.5 ％).The diagnostic accuracy of the two assays oBtained at presentation,as quantified by AUC,was no statistically differences (AUC for POCT-cTnⅠ,0.901,95％ CI,0.901 to 0.947;and for CLT-hscTnT,0.907,95％ CI,0.842 to 0.951;Z =0.235,P =0.745).The AUC for POCT-cTnI at 3 hours after admission was significantly higher than that on admission (0.931 vs.0.858;Z =-2.038,P =0.042),while there was on further improvement at 6 hours after admission (0.931 vs.0.949;Z =-1.435,P =0.151).With use of POCT-cTnI (cutoff value 0.023 ng/mL,which was the 99th percentile upper reference limit) on adimission,the clinical sensitivity was 77.5％,and the specificity was 94.2％.A single sample of POCT-cTnI at 3 hours after admission improved the diagnostic accuracy,with a sensitivity of 96.4％,a specificity of 92.0％,and a NPV of 98.6％,a PPV of 81.8％.While,with use of CLT-hscTnT (cutoff value 0.014 ng/mL,was the 99th percentile upper reference limit) at 3 hours after admission,the NPV reached to 100％.Conclusions The use of a POCT-cTnI assay in chest pain patients can identify and exclude the AMI rapidly and exactly at three hours after admission,and the diagnostic performance is equivalent to CLT-hscTnT.

Objective To study the effect of Xin Kang Injection on cardiac function of rats with adriamycin-induced myocarditis and to explore its pharmacological mechanism.Methods Seventy Wistar rats were randomly divided into seven groups:normal control group,model group,three-dosage Xin Kang Injection groups(high-,moderate-and low-dosage respectively),deslanoside group,and Huangqi group.The myocarditis rat model was established by intraperitoneal injection of adriamycin.Left ventricular end-diastolic pressure(LVEDP),left ventricular systolic pressure(LVSP),heart rate(HR),T value,and ? DP/DTmax were observed.Results LVSP,+ DP/DTmax in the treatment groups were higher and-DP/DTmax lower than those in the model group(P

Objective To describe the hemodynamic changes during piggyback liver transplantation (PBLT), and to analyze the hemodynamic correlation with various degrees of cirrhosis according to Childpugh classification. Methods Between March 1999 and June 2004, 180 patients underwent PBLT procedure in our institution, and 95 cases were selected and divided according to Child classification. The intraoperative hemodynamics of different time points were retrospectively analyzed, including mean artery pressure (MAP), heart rate (HR), central vein pressure (CVP) and mean pulmonary artery pressure (MPAP). Results Hemodynamic changes were minimal before and during anhepatic phase in all the patients. At reperfusion, a hemodynamic disturbance occurred featured by decrease of MAP and increase of MPAP. Comparison between different Child class showed that in the Child C group, MAP were lower and HR were higher before new liver phase, while CVP and MPAP were higher during new liver phase. Conclusion Hemodynamic changes were minimal before and during anhepatic phase for PBLT, while they were more severe during reperfusion, and they also correlates with the different Child class before transplantation. The more severe of the cirrhosis before transplantation according to Child classification, the greater hemodynamic changes during the operation.

Contrast-enhanced ultrasound (CEUS) is widely used to evaluate tumor microcirculation, which is useful in the differential diagnosis between benignity and malignancy. In the last 10 years, the applicability of CEUS to thyroid nodules has greatly improved due to technological refinements and the development of second-generation contrast agents. In this review, we summarize the applications of CEUS for thyroid nodules, focusing on the imaging findings of malignant and benign nodules in the existing literature and the use of those findings to predict malignancies, with an additional brief description of the utilization of CEUS for other thyroid-related diseases.
Contrast Media ; Diagnosis ; Diagnosis, Differential ; Elasticity Imaging Techniques ; Microcirculation ; Thyroid Gland* ; Thyroid Nodule* ; Ultrasonography*

Liquid-phase microextraction is a novel pretreatment technique for biological samples developed on the basis of liquid-phase extraction technology, which is simple, rapid, economical, and environmentally friendly, and has been widely used in the analysis of biological matrix samples such as blood, urine, and saliva. In this paper, we review the basic principles of the main modes of liquid-phase microextraction techniques, i.e., single-drop microextraction, dispersive liquid-liquid microextraction, and hollow-fiber liquid-phase microextraction, and the progress of their applications in biological sample pretreatment by reviewing the literature in the past five years, with a view to providing technical support and reference for sample pretreatment in the fields of in vivo drug analysis, pharmacokinetic studies and new drug development.