Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

Biopsy ; Histological Techniques ; instrumentation ; methods ; Humans ; Indicators and Reagents ; Paraffin Embedding ; Ultrasonics

OBJECTIVETo investigate the feasibility and the mid-term effects of unilateral pedicle screw fixation and transforaminal lumbar interbody fusion in treating lumbar degenerative diseases.

METHODSFrom August 2005 to May 2010, 56 patients with lumbar degenerative diseases underwent lumbar posterolateral fusion,their clinical data were retrospective analyzed. The patients were divided into two groups (unilateral group and bilateral group) according to fixation methods,27 patients in unilateral group who were underwent unilateral pedicle screw fixation, including 18 males and 9 females with a mean age of (57.5 ± 7.1) years old (ranged from 41 to 66 years); and 29 patients in bilateral group who were treated with bilateral pedicle screw fixation (on the basis of the above, with contralateral vertebral pedicle screw fixation), including 19 males and 10 females with a mean age of (54.6 ± 5.1) years old (ranged from 43 to 68 years). The clinical data such as operation time, blood loss volume, hospitalization time and cost were compared between two groups. JOA score system was used to evaluate the neurological function. And fusion status and cage-related complication were also analyzed.

RESULTSAll patients were followed up from 36 to 60 months with an average of 45.8 months. No iatrogenic nerve, blood vessels or organs injury were found during operation. Operation time, blood loss volume, hospitalization time and cost in unilateral group were better than that of bilateral group (P < 0.05). There was no significant difference in JOA score between two groups (P > 0.05). Two patients in unilateral group developed with cage related complications, 1 case was cage displacement and 1 case was cage subsidence, while 2 patients in bilateral group developed with complications of no-fusion, and there was no significant differences between two groups (P = 0.58).

CONCLUSIONUnilateral pedicle screw fixation is a satisfactory method and can obtain good effects in treating lumbar degenerative diseases in mid-term, however, the indications should be well considered.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Humans ; Intervertebral Disc Degeneration ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fusion ; methods

OBJECTIVETo investigate the effect of phosphoinositide 4-phosphate (PI4P) on human glioma U87 cells and the mechanism of action of PI4P in the development of human glioma through the overexpression or silencing of PI4P in human glioma U87 cells, and to provide a new target for basic research and clinical treatment of glioma.

METHODSLV-Helper1, LV-Helper2, pWPXLd-PI4P, and pLL3.7-shPI4P were used to package pWPXLd-PI4P and pLL3.7-shPI4P lentiviruses. The U87-GFP (PI4P-overexpression control group), U87-GFP-PI4P (PI4P-overexpression experimental group), U87-Scramble (PI4P-silencing control group), and U87-shPI4P (PI4P-silencing experimental group) cell lines were established. Wound-healing assay and Transwell assay were used to evaluate cell migration and invasion, and Western blot was used to measure the expression of PI4P in each group.

RESULTSWestern blot detected the expression of exogenous PI4P in the U87-GFP-PI4P cell line, and the U87-shPI4P cell line showed reduced expression of PI4P compared with the U87-Scramble cell line in the control group. The U87-GFP-PI4P cell line with PI4P overexpression had a significantly stronger ability of migration than the U87-GFP cell line in the control group (P<0.01); the U87-shPI4P cell line with PI4P silencing had a reduced ability of migration than the U87-Scramble cell line in the control group (P<0.01). The U87 cell line with PI4P overexpression had a significantly stronger invasion ability than the control group (P<0.05); after PI4P silencing, the experimental group showed a significant reduction in invasion ability compared with the control group (P<0.05).

CONCLUSIONSIn human glioma U87 cells, PI4P can promote the invasion and migration of glioma cells and may become a new target in the basic research and clinical treatment of glioma.

Cell Line, Tumor ; Cell Movement ; drug effects ; Glioma ; pathology ; Humans ; Neoplasm Invasiveness ; Phosphatidylinositol Phosphates ; pharmacology

Adolescent ; Child ; Collagen Type I ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Fibrillin-1 ; Fibrillins ; Heart Defects, Congenital ; metabolism ; pathology ; Humans ; Male ; Microfilament Proteins ; metabolism

With a microsystem or micropump, the release rate of drug delivery is able to be controlled easily to maintain the therapeutic efficacy. A piezoelectric membrane-valve micropump for implantable and carryhome drug delivery system is developed and tested. The influence elements of dynamic performance of the PZT actuator and valve were analyzed, and the calculation method of resonant frequency of the membrane valve was provided. Study results showed that the output performance of the micropump depended on the coupling effect of the actuator and valve. For a given actuator, the output value and the optimal frequency of a micropump could be enhanced only by valve design. Two micropumps with different valve dimensions were fabricated for comparing examination. The smaller -valve micropump obtained higher output values (the maximum flow rate and backpressure being 3.5 ml/min and 27 KPa, respectively) and two optimal frequencies (800 Hz and 3 000 Hz). The larger -valve micropump achieved lower output values (the maximum flow rate and backpressure being 3.0 ml/min and 9 KPa, respectively) and one optimal frequency (about 200 Hz). The test results suggest that the output values and optimal frequency of micropump can be improved by changing the valve dimension, and the viewpoint that checkvalve micropump works only with low acting frequency is wrong.
Drug Delivery Systems ; instrumentation ; Equipment Design ; Humans ; Infusion Pumps, Implantable ; Micro-Electrical-Mechanical Systems ; instrumentation

OBJECTIVETo investigate the effects of peroxisome proliferator-activated receptor-gamma (PPARγ) agonist rosiglitazone on the expression of cyclin D1 in lung tissue, and the proliferation of airway smooth muscle cells (ASMCs) in mice with bronchial asthma.

METHODSThirty clean BALB/c mice were randomly divided into control group (n = 10), asthma group (n = 10), and rosiglitazone treatment group (n = 10). A mouse model of asthma was established by ovalbumin (OVA) sensitization and challenge. The treatment group received rosiglitazone (5 mg/kg) by gavage 1 hour before each challenge and the control group received saline instead of OVA sensitization and challenge. Leukocytes and eosinophils in bronchoalveolar lavage fluid (BALF) were counted under a microscope. Airway structural changes were observed by hematoxylin-eosin staining. Protein and mRNA expression levels of cyclin D1 were measured by immunohistochemical staining and RT-PCR. Perimeter of the basement membrane (Pbm), total bronchial wall area (WAt), airway smooth muscle area (WAm), and number of nuclei in ASMCs (N) were determined using image analysis software, and WAt/Pbm, WAm/Pbm, and N/Pbm were calculated.

RESULTSCompared with the control group, the asthma group showed significant increases in the total number of leukocytes and percentage of eosinophils in BALF, as well as in the mRNA and protein expression of cyclin D1, but changes in these indices were significantly reduced in the rosiglitazone treatment group (P < 0.05). In addition, compared with the control group, the asthma group had significantly increased WAt/Pbm, WAm/Pbm, and N/Pbm, but rosiglitazone significantly decreased these ratios (P < 0.05).

CONCLISONSRosiglitazone may delay the process of airway remodeling by inhibiting the proliferation of ASMCs, so it can be used for preventing and treating chronic asthma.

Airway Remodeling ; Animals ; Asthma ; drug therapy ; pathology ; Bronchi ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Cell Proliferation ; Cyclin D1 ; analysis ; genetics ; Female ; Lung ; chemistry ; pathology ; Mice ; Mice, Inbred BALB C ; Myocytes, Smooth Muscle ; physiology ; PPAR gamma ; physiology ; RNA, Messenger ; analysis ; Thiazolidinediones ; pharmacology

OBJECTIVETo establish copper deficiency model in rats, and observe the damnification on internal organ of copper deficiency rats.

METHODS27 male weanling SD rats with 5-week-old were randomized into 3 groups (n = 9), i.e., control group 1, control group2 and copper deficiency group (CuD). The rats of the control group 1 were fed commercial feed with copper level of 7.0 mg/kg, the rats of the control group 2 and the copper deficiency group were fed half-purified diet with copper level of 0.73 mg/kg. In afternoons, rats were orally perfused copper sulphate solution with copper concentration 0 (control group 1), 0.133 (control group 2) and 0 mg/ml (copper deficiency group), respectively, the volume was 1% of body weight, so theory copper level of the feed in the control group2 was 11.37 mg/kg. All of rats had free access to both food and de-ionized water. Rats were sacrificed at the end of test feeding on the 42nd day, blood was sampled for analyzing the plasma ceruloplasmin activity (PCP) and its content (PPD), and erythrocyte Cu-Zn SOD (EC Cu-Zn SOD); liver was sampled for analyzing the content of Metallothionein (MT), and liver copper (LC). The organic tissues of kidney, brain, heart, liver, spleen, lung and testes are sampled for histopathologic examination.

RESULTSThe PCP and EC Cu-Zn SOD, PPD, LC and LMT of rats in copper deficiency group was significantly lower than those of rats in control group1 and control group2 (except EC Cu-Zn SOD) (P < 0.01). The cardiac muscle fibers of a part of rats in copper deficiency group were broken and eosinophilic. The endothelial cell of a coronary artery branch was presented proliferation and swelling, subendothelial space was broadened. An arteriole in the lung was showed thickening of the wall, and presented obliteration of the lumen. No obvious pathological changes of other internal organ were found.

CONCLUSIONCopper deficiency model in rats is successfully established after rats ingesting diet of low copper for 42 days. Slightly pathologic changes in the cardiovascular tissues of part of rats in copper deficiency groups are observed.

Animals ; Blood Vessels ; pathology ; Copper ; deficiency ; Deficiency Diseases ; pathology ; Disease Models, Animal ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the expression of Wnt5a gene mRNA and Wnt5a, APC, β-catenin proteins in human colorectal adenocarcinoma (CRC) and explore its clinical significance.

METHODSWnt5a mRNA level was measured in 30 patients with CRC and paired non-tumor tissues by real-time PCR. Immunohistochemical staining of Wnt5a, APC, β-catenin was performed in samples of 62 patients with CRC using SP system.

RESULTSThe relative expression level of Wnt5a mRNA in fresh CRC is 0.1232 ± 0.0140, which is significantly higher than that in adjacent colorectal mucosa (0.0497 ± 0.0074, P = 0.02). A low expression of Wnt5a protein was observed in 38 of 62 CRC. Wnt5a protein expression was closely correlated with the tumor types and the degree of tumor differentiation (P < 0.05). There was no apparent relationship with lymph node metastasis, depth of myometrial invasion and TNM stages (P > 0.05). APC protein was decreased in 38 of 62 CRC. The expression of APC was closely correlated with the tumor types (P < 0.05). There was no apparent relationship with the degree of tumor differentiation, lymph node metastasis, depth of myometrial invasion and TNM stages (P > 0.05). The expression of β-catenin was observed in cytoplasm and/or cell nuclei in 50 of 62 CRC. The positive rate of β-catenin expression was closely correlated with the degree of tumor differentiation, lymph node metastasis, depth of myometrial invasion and TNM stages (P < 0.05). There was no apparent relationship with the tumor types (P > 0.05). The expressions of Wnt5a (r = 0.271, P = 0.027) and APC (r = 0.343, P = 0.004) were correlated with that of β-catenin in CRC, respectively, but there was no correlation between the expressions of Wnt5a and APC protein (r = 0.218, P = 0.078) in the tumors.

CONCLUSIONSWnt5a, APC and β-catenin genes might be involved in the carcinogenesis and development of CRC. It is hypothesized that down-regulation of APC and Wnt5a proteins may be one of causes of ectopic expression of β-catenin in CRC.

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adenomatous Polyposis Coli Protein ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Cell Differentiation ; Colorectal Neoplasms ; metabolism ; pathology ; Down-Regulation ; Female ; Genes, APC ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Wnt Proteins ; genetics ; metabolism ; Wnt-5a Protein ; beta Catenin ; metabolism

Objective To evaluate the efficacy and safety of acute ureteroscopy for the treatment of ureterie stones during middle and late pregnancy.Methods From June 1998 to March 2005,17 pregnant women(mean age,27 years;age range,21-35 years)with ureteric stones were treated by ureteroscopy when the fetus was at 20-36 weeks of gestation(mean,29 weeks).All the cases presented with urgent symptoms such as recurrent renal colic(11 cases),fever(4)or acute obstructive anuria(2).Among 17 cases,the stones(between 6 mm?7 mm and 13 mm?21 mm)were located in the upper(8 cases),middle(5)or lower ureter(4);and on the left side(5 eases),on the right(10)and on both(2)of the lower ureter. Mild hydronephrosis were observed in 6 cases and moderate hydronephrosis in 11,Of the 17 cases,14 under- went ureteroscopic pneumatic lithotripsy;in 1 case the calculi were pushed to the renal pelvis;and 2 cases were treated by Double-J catheter drainage.Results All the urgent symptoms in 17 cases were relieved after treatment.The stone-free rate of initial treatment was 82.4%(14 of 17).Three cases with residual stones were treated by Douhle-J catheters,which were replaced every 3 months until the calculi were re- moved.No abortion,premature delivery or complications such as ureter perforation occurred.Mild renal colic occurred in 1 case after insertion of Douhle-J catheter,and it was relieved 3d later;gross hematuria occurred in l case and disappeared 6 d later without treatment.All 17 patients had normal delivery and gave birth to healthy children.Conclusions Ureteroscopy is a safe and reliable method for the treatment of ureteric calculi during middle and late pregnancy.