Objective To observe the expressions of matrix metalloproteinase-9 mRNA and the change of cerebral edema after diffuse brain injury in rats and discuss their correlation.Methods Marmaruu's diffuse brain injury model of rat was made.Real-time quantitative RT-PCR,dry-wet meth- od,histological techniques and electron microscope were used to determine the expressions of MMP-9 containing water in brain tissue and inflammatory reaction and uhrastructural changes of blood capillary at different time phases after truama.Results The expressions of MMP-9 mRNA started to increase at 1 hour,peaked at 12 hours(P

OBJECTIVESpinal muscular atrophy (SMA) is a common autosomal recessive disorder and represents one of the most common genetic causes of death in childhood. The last 10 years have seen major advances in the field of SMA, but no curative treatment is available so far. This study aimed to analyze the clinical characteristics of SMA, improve the clinical diagnosis of SMA, and explore the importance of gene diagnosis and prenatal diagnosis of SMA by gene deletion analysis.

METHODSTotally 83 cases with SMA including 55 males and 28 females were enrolled in this study. The age was between 1 day and 14 years (average 23.7 months). The clinical characteristics and changes of electromyography were assessed in all cases. The muscular biopsy was performed in 2 of 83 cases. The deletion of survival of motor neuron gene (SMN) was detected by PCR and restriction endonuclease spectrum analysis in 13 of 83 cases.

RESULTSThe 83 cases were subdivided into three clinical groups based on age of onset of symptom, age at death and achievement of certain motor milestone, 60 cases with type I, 19 cases with type II and 4 cases with type III. They were all characterized by symmetric muscle weakness (more proximal than distal) associated with atrophy, absence or marked decrease of deep tendon reflexes. Electromyographic studies showed a pattern of denervation with neither sensory involvement nor marked decrease of motor nerve conduction velocities in all cases. Muscle biopsy provided evidence of skeletal muscle denervation with groups of atrophy in 2 cases. The SMN detection revealed deletion of exon 7 and exon 8 in 11 of 13 cases, only lacking exon 7 in 1 of 13 cases and lacking exon 8 in 1 of 13 cases.

CONCLUSIONSMA is characterized by degeneration of lower motor neuron associated with muscle paralysis and atrophy. The definite diagnosis of SMA will rely on the typical clinical characteristics, changes of electromyogram and muscle biopsy and gene deletion analysis. Gene diagnosis of SMA can provide a basis for prenatal diagnosis which is of great importance in preventing SMA.

Adolescent ; Biopsy ; Child ; Child, Preschool ; Electromyography ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Muscle, Skeletal ; pathology ; Prenatal Diagnosis ; Spinal Muscular Atrophies of Childhood ; diagnosis ; genetics

After human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of exenatide for 24,48,and 72 h under the conditions of5.5 and 33 mmol/L glucose,the cell viability was detected by MTF assay.The apoptosis rate of endothelial cells was measured by flow cytometry.Protein kinase B (Akt) phosphorylation,Bcl-2,and Bax protein expression were detected by Western blotting.The results showed that the cell viability was decreased after HUVECs were incubated with high glucose for48 and 72 h (P＜0.01).Treatment with 1,10,and 100 nmol/L exenatide for48 h significantly increased the cell viability in the presence of high glucose,in a dose-dependent manner (P＜0.01).High glucose inhibited Akt phosphorylation and Bcl-2 protein expression,stimulated Bax protein expression,with decreased Bcl-2/Bax ratio and increased apoptosis ratio of cells.After treatment with exenatide,the cell apoptosis reduced,Akt phosphorylation and Bcl-2 protein expression increased,and Bax protein expression decreased,with increased Bcl-2/Bax ratio (P ＜ 0.01).These effects of exenatide on endothelial cell were abrogated by an inhibitor of phosphotylinosital 3 kinase (PI3K) LY294002 (P ＜ 0.01),suggesting that the exenatide may regulate Bcl-2/Bax protein expression and inhibit endothelial cell apoptosis induced by high glucose through PI3k/Akt signal pathway,and thus may protect endothelial cells.

Objective To explore the diagnostic significance of differential cell count in induced sputum to chronic cough and assessment of airway inflammation.Methods The sputum of 335 chronic cough patients were induced.Differential cell counts were measured in these samples.The side effects were observed during the induced procedure.The final diagnosis was made based on clinical manifestation and examination findings including pulmonary function tests,provocation test,induced sputum cell differentials, etc.Results The cause of chronic cough was defined in 322 patients.The six most important causes of cough were typical asthma(TA,n=84),eosinophilic bronchitis (EB,n=62),atopic cough (AC,n= 42),cough variant asthma (CVA,n=40),gastroesophageal reflux cough(GERC,n=37),rhinitis and/ or paranasal sinusitis (PNDs,n=32),and others and indefinite cause (n=25,13).Percentage of eosinophils were significantly increased in the induced sputum of AC,EB,CVA,and GERC patients (0.005,0.052,0.059,0.234) compared with those in other causes and the healthy controls (0) (P

OBJECTIVETo evaluate the effects of asthma and inhaled corticosteroids (ICS) in children on the final adult height.

METHODSA search was performed to collect studies evaluating the relationship between asthma and ICS in children and the final adult height in PubMed, BCI, EMbase, Web of Science, CNKI and Wanfang databases, then a systemic review and Meta analysis were conducted.

RESULTSSix studies evaluating the relationship between childhood asthma and the final adult height were enrolled. Three of them indicated that the final adult height was not influenced by childhood asthma. Two of them suggested a mild effect, and the effect was correlated with severity of childhood asthma. One of them indicated that a lower final adult height related to childhhod asthma was found only in black females without a high school education. Four studies evaluating the relationship between ICS and the final adult height were included. Compared with the non-ICS treatment group, healthy control group and the target height, ICS treatment had no effects on the final adult height.

CONCLUSIONSChildhood asthma does not or only mildly decrease the final adult height. ICS treatment does not significantly affect the final adult height.

Administration, Inhalation ; Adrenal Cortex Hormones ; adverse effects ; Adult ; Asthma ; drug therapy ; Body Height ; drug effects ; Child ; Humans

Objective To explore the chronic effects of mild and moderate iodine excess and iodine restriction on apoptosis of thyrocytes. Methods Wistar rats were exposed to 4 different doses of iodine: 4 μg/d (control), 6 μg/d (1.5 fold iodine excess), 12 μg/d (3 fold iodine excess), and 24 μg/d (6 fold iodine excess) for 1, 2, 4 and 8 months. Some rats treated for 8 months were fed with 4 μg/d iodine for another 3 months. Urinary iodine concentration was monitored by arscnic/cerium catalyzing spectrophotography. Apoptosis was determined by flow cytometry after Annexin V-FTTC staining and uhrastructure assessment under electronic microscope. Cell cycle kinetics was analyzed by flow eytometry after propidium iodine staining. Fluorescent measurement by DCFH-DA probe was used to determine the intracellular reactive oxygen species (ROS) level. Expressions of apoptic proteins were analyzed by flow cytometry and immunohistochemistry. Results Apoptotosis rate and ROS production in thyrocytes were significantly increased in 3 and 6 fold iodine excess groups after 4 months and 8 months (all P < 0.05), which was reversed with iodine restriction. 6 fold iodine exposure was proved to cause a reduction of cells in GOG1-phase (64% and 67% vs 80%, both P < 0. 05) and a concomitant accumulation in S-phase (5% and 6% vs 3%, both P <0.05) after 4 months and 8 months. Expressions of Fas, FasL and TRAIL proteins in 3 and 6 fold iodine excess groups after 8 months were increased by 2 to 4 times compared with control group and did not return to normal after iodine restriction. Bcl-2 and Bax remained constant. Positive correlations were observed among iodine amount, apoptosis rate and ROS level in 6 fold iodine excess group after 8 months (r = 0. 637-0.790, P < 0.01). Conclusion Chronic iodine excess results in thyrocyte apoptosis due probably to generation of ROS.

Objective To construct four types of glucagon-like peptide-1 (GLP-1) and human serum albumin (HSA) fusion proteins that can be realeased at different rate in vivo by introducing protease cleavage sites between these two moieties.The therapeutic effect and release rate are studied to achieve balanced pharmacokinetics ( PK) and pharmacody-namics ( PD) of GLP-1 and HSA fusion proteins.Methods The gene with different polypeptide joint of GLP-1 and HSA fusion proteins were synthesized by overlap extension PCR amplification, cloned into expression vector pPIC9 and transformed into Pichia pastoris GS115.Then, fusion proteins were obtained by protein purification after being induced by methanol.The preliminary PK and PD of the fusion proteins were studied after purification.Results The fusion protein Gly2-GLP-1-GGGGG-HSA showed no release while Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA showed a slow, medium and fast release rate, respectively, after incubation with furin.In vitro biological activity test results dispalyed that each type of fusion protein promoted insulin secretion of MIN6 cells.In vivo PK test indicated the half-life size of fusion proteins was the largest in Gly2-GLP-1-GGGGG-HSA, followed by Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA.In vivo PD test exhibited hypoglycemic activity that was the highest in Gly2-GLP-1-VTR-HSA, followed by Gly2-GLP-1-SARSVRA-HSA, Gly2-GLP-1-GRSRVTRSV-HSA, and Gly2-GLP-1-GGGGG-HSA.Conclusion GLP-1 can be released from fusion proteins with full activity after the introduction of protease cleavage sites.Releasable fusion proteins at an appropriate release rate have the most balanced PK and PD.

Objective To investigate the effects of standardized tertiary rehabilitation (STR) on limb motor function (LMF) after stroke.Methods Eighty-two patients were divided into a primary cerebral infarction group (PCI group) and a primary cerebral hemorrhage group (PCH group),and then randomly further divided into experi- mental and control sub-groups.All patients received routine internal medicine treatment,supplemented with stand- ardized tertiary" rehabilitation in the experimental groups.All patients were assessed with the simplified Fugl-Meyer motor function assessment (S-FMMFA) at enrollment,and 1,3 and 6 months after their stroke.Results The scores of the experimental groups were higher than those of the controls.The experimental groups scores were 26.10% of normal at the time of the enrollment,and improved to 42.52%,65.62% and 83.71% by the end of the 1st,3rd and 6th month,respectively.The control groups started at 18.51%,and progressed to 24.85% ,37.24% and 45.84% over the same interval.Conclusion STR was associated with improved LMF scores of stroke pa- tients.

OBJECTIVETo investigate the expression features of glypican-3 (GPC-3) and its diagnostic and differential values in hepatocellular carcinoma (HCC).

METHODSRat hepatoma models were made and the dynamic expression features of GPC-3 protein and its gene were investigated by Western blotting and RT-PCR respectively. Liver specimens from 36 HCC patients were collected by self-control method and the expression and clinicopathological features of GPC-3 were analyzed by immunohistochemistry. Serum GPC-3 levels were quantitatively detected by ELISA and its efficiency for HCC diagnosis was evaluated in patients with liver diseases.

RESULTSThe incidence of GPC-3 was 0% in control, 83.3% in degeneration, 100% in precancerosis and 100% in canceration during dynamic formation of rat hepatoma, respectively. The positive GPC-3 was brown granule- like staining localized in membrane and cytoplasm in human HCC.

CONCLUSIONSThe GPC-3 positive rates were 80.6% in HCC, 41.7% in surrounding tissues and none in distal tissues (P < 0.01), respectively. No positive relationship presented between GPC-3 and differentiation grade or the number of tumor except of tumor size (Z = 2.941, P < 0.01). The incidence of serum GPC-3 was 52.8% in HCC patients except of one patient with cirrhosis. No significant differences were found between GPC-3 and sex, age, AFP, tumor number, Child classification or extrahepatic metastasis except of tumor size (χ² = 6.318, P < 0.05) and HBV infection (χ² = 23.362, P < 0.01). Combined detection of GPC-3 and AFP could rise up diagnosis of HCC. GPC-3 expression closely associated with HCC and might be useful for early diagnosis of HCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glypicans ; metabolism ; Humans ; Liver ; pathology ; Liver Neoplasms ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Rats ; Rats, Sprague-Dawley ; Young Adult

OBJECTIVETo explore the impact of different warm ischemia time on structure and function of the non-heart-beating donor lung and to find out the feasibility of non-heart-beating donor in rat lung transplantation.

METHODSSixty Sprague-Dawley rats were randomly divided into 3 groups: heart-beating donor (HBD) group, non-heart-beating donor (NHBD) with 30 minutes of warm ischemia time (WIT) group and NHBD with 60 minutes of WIT group. Each group has 10 pairs (the donors and the recipients). The donor lungs of group HBD were flushed with low potassium dextran (LPD) solution at 4 degrees C after asystolia while the lungs of group NHBD-30 and group NHBD-60 remained ventilated at the room temperature for 30 and 60 minutes after asystolia and then were flushed with LPD solution. All the donor lungs remained inflated when they were stored in LPD solution at 4 degrees C for 4 hours. The recipient rat underwent left thoracotomy, and then orthotopic left lung transplantation. Followed by a right thoracotomy, the right pulmonary hilum were ligated with one-hour reperfusion and ventilation.

RESULTSAll the recipients in group HBD and group NHBD-30 survived the observation period of one hour with excellent gas exchange, whereas 4 of recipients in group NHBD-60 survived for 10 minutes after the ligation of right pulmonary hilum and 3 for 20 minutes. The pulmonary compliance, ultrastructure, energy metabolite and other markers revealed no significant differences between group HBD and group NHBD-30 (P > 0.05). But the differences between group NHBD-60 and other two groups were significant (P < 0.05).

CONCLUSIONSThe adoption of non-heart-beating donor could be a safe and effective method to expand the lung donor pool. The NHBD lung with 30 minutes of WIT may be suitable for lung transplantation in rat.

Animals ; Heart ; physiopathology ; Ischemia ; physiopathology ; Lung ; physiopathology ; ultrastructure ; Lung Transplantation ; Male ; Microscopy, Electron ; Models, Animal ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Tissue Donors