Objective To observe the effects of glucagon like peptide-1 (GLP-1) analogues liraglutide on expressions of nitric oxide synthase (NOS) and cyclo-oxygen-ase (COX)2 in renal medulla of type 2 diabetes rats, and the mechanism of its lowering blood pressure and promoting excretion of water and salt in kidney. Methods Type 2 diabetes model rats were generated by high-fat and high-sugar feeding for 8 weeks followed by intraperitoneal injection of streptozotocin (STZ). Subse?quently, eighteen type 2 diabetes rats were divided into two groups: liraglutide treatment group (DMT) and diabetes group (DM). Twelve normal rats were divided into two groups: liraglutide treatment wild type group (WTT) and wild type group (WT). DMT and WTT groups were given liraglutide (200μg/kg) by subcutaneous injection, DM and WT groups were given equivalent normal saline by the same way. The levels of blood glucose and blood pressure were detected at 0, 2, 4 and 6 weeks after treatment in groups of rats. Samples of urine were collected for detecting ion concentrations (K+, Na+and Cl-) af?ter treatment for six weeks. Rats were sacrificed and blood samples were collected for detecting ion concentrations (K+, Na+and Cl-). The expression levels of NOS and COX2 mRNA and protein in renal medulla were detected by real-time PCR and Western blot assay. Results After treating with liraglutide, the values of blood glucose (F=5.933, P<0.05) and blood pres?
sure (F=22.070, P<0.05) were gradually decreased in DMT group. After treatment with liraglutide for 6 weeks, the values of blood glucose (mmol/L:12.78 ± 3.82 vs. 18.75 ± 1.68) and blood pressure (mmHg:119.98 ± 4.43 vs. 136.42 ± 4.48) were signifi?cantly decreased (P<0.05) in DMT group than those of DM group (P<0.05). There were no significant differences in the concentrations of K+, Na+and Cl-between the two groups. There were higher levels of K+(mmol/L:46.55 ± 6.43 vs. 33.13 ± 9.71), Na+(mmol/L:56.33±8.83 vs. 41.20±7.25) and Cl-(mmol/L:159.81±25.06 vs. 71.44±12.99) in urine in DMT group than those of DM group (P<0.05). The mRNA levels and protein expressions of NOS and COX2 in renal medulla were significant?ly increased in DMT group than those of DM group (P<0.05). Conclusion GLP-1 analogues liraglutide may enhance the expression of COX2 by increasing the expression of NOS to excrete water and salt, and decrease blood pressure.

Objective To study correlation between systemic sclerosis(SSc)and plasma D-dimer and to reveal the probably rules of the fibrinolytic systems in SSc.Methods In 2013 January to 2014 January,cellected 32 patients with SSc and 35 with healthy controls to detected level of plasma D-dimer.Logistic and t student test were used for statistical analysis for correlation between SSc and pulmonary artery hypertension.Results When compared to healthy controls (0.28±0.04μg/ml),the level of plasma D-dimer were significantly increased in SSc patient (0.31±0.05μg/ml,t=1.997,P=0.008).After stratifying SSc patients according to disease subset,whereas patients with diffuse subset displayed substantially increased values (0.41±0.06μg/ml,t=2.051,P<0.001).The level of plasma D-dimer was associated with pulmonary artery hyper-tension (OR=4.38,95%CI=2.59~8.91,P=0.008).Conclusion Demonstrated that SSc patients with diffuse subset are characterized by increased plasma D-dimer values,reflecting a potential activation of fibrinolytic cascaded,which might finally predispose these patients to thrombotic complications and pulmonary artery hypertension.

Objective To investigate the values of three-dimensional visualization technology in the diagnosis and surgical treatment of hepatic hemangioma. Methods Thirty two patients with hepatic hemangioma who had been hospitalized during the period from July 2010 to January 2014 in our hospital were scanned by 64-slice spiral computerized tomography (CT) before surgical treatment. Three-dimensional (3D) reconstruction based on the CT data was carried out to achieve dignosis and surgical planning. Assisted with the 3D model , we chose the best surgical procedure for liver resection, protecting the major blood vessels near hemangioma and retaining normal liver tissue as far as possible. Results The location, size and shape of hepatic hemangioma, vascular variation, and spatial relationship with intrahepatic vessel were shown factually by three-dimensional reconstruction. All the hemangiomas were preoperatively assessed to be resectable. The compliance rate for preoperative surgical planning to actual surgery was 100%. Under assistance of the 3D model during surgery , 14 patients received laparoscopic hepatectomy and 18 underwent hepatectomy. Pringle′s maneuver was applied in 18 patients , with blocking time of (15.32 ± 7.12) min and blood loss of (188.63 ± 66.37) mL. The postoperative complications included pleural effusion in 5 patients and incision infection in one patients. Conclusions Three-dimensional visualization technology for the individualized diagnosis and treatment of hepatic hemangioma helps reduce surgical trauma and incidence of postoperative complications.

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

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

OBJECTIVETo investigate the potential role of serum citrulline level in evaluating the intestinal absorptive area and capacity in patients with short bowel syndrome (SBS).

METHODSSerum citrulline concentration was determined using high performance liquid chromatography (HPLC) in SBS patients (n=22) and healthy controls (n=33). In SBS patients, the remnant small bowel lengths and diameters were measured by radiography, and their 5- hour urine D- xylose excretion and intestinal protein absorption were also determined. The correlationship of serum citrulline level with remnant small bowel length, surface area, protein and D- xylose absorption was analyzed. The 6 patients receiving intestinal rehabilitative therapy, serum citrulline level, protein and D- xylose absorption after therapy were also measured.

RESULTSSerum citrulline level of SBS patients was significantly lower than that of healthy controls [(5.94+/- 2.65) vs [(16.87 +/- 5.97) micromol/L, P < 0.01]. In SBS patients, serum citrulline was positively correlated with remnant small bowel length and surface area (r=0.82 and r=0.86 respectively). There was also a significant correlationship of serum citrulline level with 5- hour D- xylose excretion (r=0.56) and intestinal protein absorption (r=0.48). Serum citrulline, 5- hour D- xylose excretion and intestinal protein absorption were all significantly raised in patients after rehabilitative therapy, although no correlation of increasing percentage was found among above three parameters.

CONCLUSIONSSerum citrulline concentration is positively correlated with intestinal absorptive area and capacity in SBS patients. It is a potential marker for evaluating the severity of intestinal failure and the efficacy of rehabilitative therapy in short bowel patients.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Citrulline ; blood ; Female ; Humans ; Intestine, Small ; metabolism ; Male ; Middle Aged ; Short Bowel Syndrome ; blood ; metabolism ; physiopathology ; Xylose ; metabolism ; Young Adult

Objective:To establish an HPLC method for the determination of four kinds of isoflavone compounds including daid-zin, tectoridin, daidzein and tectorigenin in the extracts of Pueraria lobata flowers. Methods:The separation was performed on an Agi-lent C18 column(250 mm × 4. 6 mm, 5 μm) using a mobile phase of methanol-acetonitrile-water(2∶1∶2). The flow rate was 1. 0 ml· min-1 ,and the detection wavelength was 264nm. The column temperature was 25℃ and the sample size was 20 μl. Results:The de-tection could be accomplished within 10 minutes with good separation and specificity of four isoflavone compounds with the retention timeof3.4,3.8,5.7and7.2min,respectively. Thelinearrangewas0.784-78.440 μg·ml-1(daidzin),2.000-200.000 μg· ml-1(tectoridin) and 0.800-80.020 μg·ml-1 (daidzein and tectorigenin),and the relative coefficient was 0.999 9, 0.999 8, 0. 999 7 and 0. 999 9, respectively. The average recovery was 100. 54%(RSD=1. 66%,n=6),100. 03%(RSD=1. 00%, n=6), 99. 48%(RSD=1. 76%, n=6) and 100. 92%(RSD=2. 26%, n=6), respectively. Conclusion: The method is simple, rapid and accurate with good repeatability, which can be used in the rapid determination of isoflavone compounds in the flowers of Pueraria loba-ta.

OBJECTIVETo explore the mechanism of rehabilitation after middle cerebral artery occlusion (MCAO).

METHODSMCAO model was reproduced with two-kidney, two clip renovascular hypertensive rats stroke-prone (RHRSP), which were divided into two groups, the treated group (treated with electric stimulus) and the control group (untreated model) randomly. The rehabilitation of rats was evaluated by balance beam walking test. The ultrastructural changes of neurons and astrocytes, expressions of glial fibrillary acidic protein (GFAP)-positive cells, neurofilament (NF) protein, and cerebral capillary dilatation M-associated protein-2 (MAP2), as well as the neurons apoptosis and the number of dilatation of cerebral capillary in the margin of infarcted area were observed by the end of 1st, 3rd, 6th and 9th week after modeling.

RESULTSThe motor function of paralysed limbs recovered better in the treated group than that in the control group by the end of 3-9th week after MCAO, the expression of GFAP-positive cells in astrocytes and NF, MAP2 in neurons as well as the number of cerebral capillary dilatation at the margin of infarcted area were higher than those in the control group (P < 0.05).

CONCLUSIONElectric stimulation treatment could improve the recovery of motor function of paralyzed limbs. It might be due to the effect of electric stimulus in increasing astrocytes proliferation, reinforcing activity of neurons and evoking the dilatation of cerebral capillary.

Animals ; Apoptosis ; Astrocytes ; ultrastructure ; Electric Stimulation Therapy ; Glial Fibrillary Acidic Protein ; metabolism ; Infarction, Middle Cerebral Artery ; pathology ; physiopathology ; rehabilitation ; Male ; Neurons ; ultrastructure ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To optimize the extraction technology of flos Puerariae lobata isoflavone .Methods The flos Pu-erariae lobata isoflavone was distilled by ethanol circumfluence .Total flavonoids ,tectoridin and tectorigenin extracted from Puerariae using the UV and HPLC spectromertry methods were taken as evaluation indexes .Extraction technology was opti-mized with L9 (34 ) orthogonal test on the base of single observation of ethanol concentration ,solvent dosage and distilling time . Results The best extraction technology of flos Puerariae lobata isoflavone was :to add 12 times the amount of 70% ethanol for 90 minutes for the first time ,and 10 times the amount of 70% ethanol for 60 minutes for the second time .Conclusion The op-timized extraction process of flos Puerariae lobata isoflavone is reasonable and feasible ,and it can offer reference to actual pro-duction .

Objective To construct an expression system to produce the virus-like particles containing a part of the sequence of PSA mRNA, which are ribonuclease-resistant due to the encapsulation of the mRNA by bacteriophage MS2 coat proteins. Methods The PCR products of PSA cDNA fragments were cloned to TA vector pBS-T, then the targeted segments could be obtained when the pBS-T-PSA were digested by restriction endonuclease Hind Ⅲ and cloned to prokaryocytic expression vector pNCCL1. The recombinant plasmids named PNCCL1-PSA were transfected into E. Coli BL21-DE3 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. The bacteriophage MS2 coat protein which expressed in BL21 can self- assemble to form ribonuclease resistant virus-like particles and the PSA mRNA was encapsulated into virus-like particles. Conclusions The virus-like particle containing PSA mRNA can be expressed in prokaryocyte and it can be used as standard and control in detecting PSA mRNA. It provides a new, stable and ribonuclease-resistant RNA standard in RNA detection.