Objective: To investigate the subcellular localization of a sesquiterpene synthase from Aquilaria sinensis (ASS) and compare three transient expression systems in the subcellular localization study. Methods: An ASS gene was amplified by RT-PCR using specific primers and cloned into the pEZS-NL and pFGC5941GFP to generate two plant expression vectors: pEZS-NL-ASS- GFP and p5941-GFP-ASS. Three transient expression systems, agroinfiltration of tobacco leaves, particle bombardment of onion epidermal cells, and PEG transformation of protoplasts isolated from ASS calli were adopted and compared. The expression of the GFP fusion proteins were observed by a confocal laser scanning microscopy. Results: The green fluorescence could be observed in all the three systems. However, the results were comparatively different. The cytoplasm and plastid localization of the GFP fusion protein observed in protoplasts was comparatively clear and was consistent with the studies in other plant species. Conclusion: The results demonstrate that the ASS is located in the cytoplasm and plastid in the ASS protoplasts and the different locations in three systems might be caused by the distinct characters of the heterologous or homologous cells.

Objective To investigate the expressions of serum cystatin C CysC),retinolbinding protein (RBP),serum and urinary β2-microglobulin (β2-MG) and their effects on predicting renal damage in primary hypertension in elderly patients.Methods A total of 149 patients were divided into 3 groups of hypertension grade 1 (n =50),hypertension grade 2 (n =50) and hypertension grade 3 group (n=49).92 healthy controls who took physical examination were selected as control group.The levels of serum CysC,RBP,serum and uric β2-MG,blood urea nitrogen,creatinine and other biochemical indexes were detected.The complications of hypertension were observed.The differences in renal function indexes between groups and their correlations were analyzed.Results There were 86 cases with hyperlipidemia (57.7％),71 cases with atherosclerosis (47.7％),59 cases with type 2 diabetes mellitus (39.6％),56 cases with coronary heart disease (37.6％),and 49 cases with cerebral vascular disease (32.9％) in 149 hypertensive patients.The levels of serum CysC,RBP,serum and uric β2-microglobulin were increased in hypertensive patients as compared with healthy controls (P＜0.05 or 0.01),and the extents of increments in the above indexes were positively correlated with blood pressure level [CysC:(1.2± 0.5) mg/L,(1.7±0.6) mg/L,(2.2±0.9) mg/L;RBP:(53.1±10.2) mg/L,(65.2±16.3)mg/L,(75.8±17.5) mg/L;serum β2-MG:(2.7±1.5) mg/L,(4.0±2.1) mg/L,(4.9±2.0) mg/L;uric β2-MG:(178.3±73.9) ng/L,(237.9±78.6) ng/L,(277.5±87.5) ng/L;respectively,P＜0.05or 0.01].Conclusions Elderly hypertensive patients are complicated with a variety of diseases.The levels of serum CysC,retinol-binding protein,serum and uric β2 microglobulin have significant diagnostic values on renal impairment caused by hypertension,and can be considered as sensitive factors for early renal impairment in essential hypertension.

AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P ＜ 0.05).The positive rates of the cells before and after sorting were significantly different (P ＜0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.

Objective: To evaluate the clinical efficacy of sinew-regulating bone-setting manipulations plus exercise therapy in treating chronic non-specific low back pain (CNLBP). Methods: A total of 65 CNLBP patients were divided into two groups by the random number table method. Thirty-three cases in the treatment group were intervened by sinew-regulating bone-setting manipulations plus exercise therapy; 32 cases in the control group were intervened by medium-frequency electrotherapy plus exercise therapy. Before and after treatment, visual analog scale (VAS), dynamic and static muscle endurance of low back, median frequency (MF) of surface electromyography (sEMG) and Oswestry disability index (ODI) were used to evaluate the low back function. The therapeutic efficacy was estimated after treatment. Results: The two groups each had 2 dropouts during the study. The total effective rate was 90.3％ in the treatment group versus 66.7％ in the control group, and the between-group difference was statistically significant (P<0.05). After treatment, the VAS score, dynamic and static muscle endurance of low back, MF of sEMG and ODI score all changed significantly in both groups (all P<0.05); all the items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Sinew-regulating bone-setting manipulations plus exercise therapy can effectively release pain in CNLBP patients, increase muscle endurance of the low back and improve the quality of life, and its therapeutic efficacy is more significant than that of medium-frequency electrotherapy plus exercise therapy.

Objective To investigate the blood pressure status of preschool children in urban areas of Qingdao, and to determine the relationship between obesity and blood pressure in preschool children. Methods A stratified cluster sampling method was used to select a total of 13 kindergartens in urban districts of Qingdao. Height, weight, waist circumference, hip circumference and blood pressure of children in three classes were measured. Body mass index (BMI), waist to hip ratio and waist to height ratio were calculated and the relationship between obesity and blood pressure was analyzed. Results The mean values of systolic and diastolic blood pressure in preschool children in urban areas of Qingdao were (95.52±7.66) and (62.78±6.52) mmHg, respectively.The detection rate of hypertension in preschool children was 13.50%. The SBP and DBP were positively correlated with BMI, waist circumference, hip circumference and waist to height ratio. There was a linear regression relationship between body mass index and age and blood pressure. The risk of hypertension in overweight and obese children was 5.191 and 2.824 times of normal body weight, respectively. Conclusions The prevalence of hypertension in preschool children in Qingdao urban areas is high.Overweight and obesity are risk factors for elevated blood pressure.Therefore, while preventing preschool children from obesity, preschool children's blood pressure monitoring and blood pressure monitoring and early intervention of hypertension of preschool children should be implemented.

Periventricular white matter damage is one of the characteristics of brain damage in preterm infants, and it is the most important type of encephalopathy. The pathological changes including the white matter of coagulation necrosis, oligodendrocyte damage, myelin damage, axonal injury and reactive gliosis and microglia infiltration in necrotic areas. All of these lesions are closely related to the nervous system sequelae in later-neonatal period. The pathogenesis of periventricular leukomalacia in premature infants are mainly cause by its immature brain vascular, and precursor oligodendrocytes of the attack of hypoxia, ischemia, infection, oxygen free radicals, inflammatory cytokines, increasing glutamate, and other high-risk factors. In this paper, an overview of progress in the study of the pathogenesis of periventricular white matter damage in premature infants through literature review to provide a theoretical support for clinical prevention, diagnosis and treatment.
Apoptosis ; Cerebrovascular Circulation ; Cytokines ; physiology ; Excitatory Amino Acids ; toxicity ; Humans ; Infant, Newborn ; Infant, Premature ; Leukomalacia, Periventricular ; classification ; etiology ; Risk Factors

Objective To evaluate effects of interleukin-36α (IL-36α) on psoriasiform skin lesions and C-C motif chemokine ligand 20 (CCL20) expression in mice.Methods Totally,30 BALB/c female mice were randomly and equally divided into 3 groups:control group treated with topical vaseline cream on the shaved back and intracutaneous injection with phosphate buffer saline (PBS),model group treated with topical imiquimod cream on the shaved back and intracutaneous injection with PBS,experimental group treated with topical imiquimod cream on the shaved back and intracutaneous injection with IL-36α solution.Psoriasis area severity index (PASI) was used to evaluate changes of psoriasiform skin lesions in mice,and light microscopy to observe morphological changes of skin lesions and to measure the thickness of the epidermis.Real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the expression of IL-36α in skin lesions in the control group and model group,and qRT-PCR,Western blot analysis and immunohistochemical study to evaluate changes of CCL20 levels in skin lesions.Results The model group showed significantly increased mRNA (△ Ct value:0.0195 ± 0.0059) and protein expression (3.922 ± 0.248) of IL-36α compared with the control group (mRNA:0.0012 ± 0.0004,P ＜ 0.05;protein:0.690 ± 0.025,P ＜ 0.05).The mRNA and protein expression of CCL20 were significantly higher in the experimental group than those in the model group (mRNA:2.152 ± 0.793 vs.0.999 ± 0.178;protein:0.397 ± 0.033 vs.0.145 ± 0.030;both P ＜ 0.05),and higher in the model group than those in the control group (mRNA:0.378 ± 0.075;protein:0.025 ± 0.009;both P ＜ 0.05).Immunohistochemical study showed that the expression intensity of CCL20 in skin lesions significantly increased in the experimental group compared with that in the model group (Z =2.294,P ＜ 0.05).Conclusion IL-36α may aggravate psoriasiform skin inflammation in mice by promoting CCL20 expression.

?AlM: To investigate the expression of miR-181 in the lens tissue of cataract and the regulating mechanism of miR-181 on apoptosis of human lens epithelial cell.
?METHODS:Real time q-PCR was used to measure the expression of miR-181 in the anterior lens capsules of age - related cataract and human lens epithelial cell apoptosis model. miR- 181 mimic and inhibitor were transfected using Lipofectamine 2 000 to regulate the expression of miR-181, and then Real time q-PCR was used to verify transfection efficiency. Flow cytometry was used to detect the change of cell apoptosis rate.
? RESULTS: Compared with control group, the expression of miR-181 was significantly higher in both the anterior lens capsules of age-related cataract and human lens epithelial cell apoptosis model; the relative expression of miR-181 in lens epithelial cells transfected with miR-181 mimic was increased, whereas decreased in cells transfected with miR-181 inhibitor;the apoptosis rate of cells transfected with miR - 181 mimic was increased, while reduced in miR-181 inhibitor group. Each result was statistically significant (P<0. 01).
?CONCLUSlON:High expression of miR-181 is detected in anterior lens capsule of age-related cataract. miR-181 might play a certain role in the pathogenesis of cataract via promoting human lens epithelial cell apoptosis. miR-181 probably becomes a new approach for the nonoperative treatment of cataract, but the concrete mechanism still needs to be further studied.

Objective To evaluate the efficacy of emergency endoscopic intervention in acute obstructive suppurative cholangitis (AOSC) complicated with septic shock.Methods A total of 54 patients with AOSC and septic shock who underwent therapeutic emergency ERCP were included in this retrospective study,and were evaluated by the shock index (SI).Results ERCP was performed for all patients in 24hours after hospitalization,and the average ERCP operation time was 23.8 ± 12.5 min.All 54 patients underwent EST,46 of whom received ENBD,7 biliary stenting and 1 transferred to surgery due to bleeding.The post-ERCP mortality rate was 0,and the complications included 1 case of pancreatitis and 2 cases of pneumonia.The positive rate ofGram-Negative bacillus before ERCP was 46.9％ (15/32).The SI before ERCP was 1.250 ±0.200,which decreased to 0.950 ±0.119 at 2hr after the procedure (P ＜0.001 ),and decreased further to 0.598 ± 0.099 ( P ＜ 0.001 ) at 24 hours after ERCP.Conclusion Therapeutic emergency ERCP is of great importance in the treatments for AOSC complicated with septic shock.

This study aimed to explore the impact of depression caused by chronic unpredictable mild stress (CUMS) on in vivo activity of six kinds of CYP450 isoforms in rats. According to 'Katz' method, the model of CUMS was established. Tolbutamide, chlorzoxazone, theophylline, midazolam, omeprazole and dextromethorphan were chosen as probe substrates of CYP2C6, CYP2E1, CYP1A2, CYP3A2, CYP2D1 and CYP2D2 of rats. Plasma concentration of six kinds of CYP450 in control group and model group were determined by LC-MS/MS and computed pharmacokinetic parameters. Consequently, metabolism of theophylline and chlorzoxazone accelerated significantly (P < 0.01), but tolbutamide, dextromethorphan, omeprazole and midazolam had no significant difference. The present study proved that depression caused by CUMS had strong induction to CYP1A2 and medium induction to CYP2E1.
Animals ; Chlorzoxazone ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 Enzyme System ; metabolism ; Depression ; Dextromethorphan ; metabolism ; Liver ; enzymology ; Midazolam ; metabolism ; Omeprazole ; metabolism ; Rats ; Stress, Physiological ; Tandem Mass Spectrometry ; Theophylline ; metabolism ; Tolbutamide ; metabolism