Objective To investigate the effect of dichloroacetate on the expression of Kv1.5 in a rat model of pulmonary arterial hypertension (PAH) .Methods Thirty-two male SD rats weighing 200-250 g were randomly divided into 4 groups ( n = 8 each): normal control group (group C), dichloroacetate control group (group D),PAH group, and PAH + dichloroacetate group (group PD). PAH was induced by left lung resection combined with subcutaneous injection of monocrotaline 60 mg/kg in PAH and PD groups. In group PD, dichloroacetate 80 mg/kg was given through a gastric tube into stomach once a day for 28 consecutive days after monocrotaline injection,while the equal volume of normal saline was given instead of dichloroacetate in group PAH. Group D only received dichloroacetate 80 mg/kg through a gastric tube into stomach once a day for 28 consecutive days. Pulmonary arterial pressure (PAP) was measured at day 28 after monocrotaline injection. The rats were then sacrificed and lung tissues were removed to calculate the percentage of thickness of the tunica media of pulmonary artery and right venicular hypertrophy index and to determine the proliferating cell nuclear antigen (PCNA) and Kv1.5 protein expression (by Western blot) and Kv1.5 mRNA expression (by RT-PCR).Results Compared with group C, the PAP,percentage of thickness of the tunica media, right ventricular hypertrophy index were significantly increased, Kv1.5 mRNA and protein expression was down-regulated and PCNA expression was up-regulated in groups PAH and PD ( P ＜ 0.05). Compared with group PAH, the PAP, percentage of thickness of the tunica media, right ventricular hypertrophy index were significantly decreased, Kv1.5 mRNA and protein expression was up-regulated and PCNA expression was down-regulated in group PD (P ＜ 0.05). There was no significant difference in the indexes mentioned above between group C and group D ( P ＞ 0.05). Conclusion Dichloroacetat alleviates PAH through upregulating Kv1.5 expression in lung tissues and inhibiting pulmonary vascular remodeling in rats.

Objective To explore the underlying mechanism of potential effect of sildenafil on porcine pulmonary artery smooth muscle cells (SMCs) proliferation induced by serotonin. Methods Porcine pulmonary artery SMCs at 3-5 passages were randomly divided into 4 groups: control group (CON), 5-HT group (HT), sildenafil group (SIL) and sildenfil-5HT combined group (S-HT). Pulmonary artery SMCs at exponential growth phase were serum starved with 0.2% FBS for 72 h, followed by sterile PBS, serotonin (10 μmol/L) and sildenafil (1 μmol/L) incubation in CON group, HT group and SIL group, respectively. In combined group (S-HT): pulmonary artery SMCs were serum starved and then exposed to sildenafil for 20 min, followed by serotonin incubation for indicated time. The phosphorylation of extracellular signal regulated kinase (ERK1/ERK2) was measured by Western blot at indicated time points. Flow active cell sorting (FACS) was used to evaluate the ratio of S phase cells in the cell cycle after 24 h of treatment. MTT color metric method was used to analyze SMCs proliferative index after 72 h of treatment. Results Compared with CON group, the phosphorylation of ERK1/ERK2, the percentage of cells in S phase, and the cell proliferation index increased remarkably after incubation with 5-HT (P<0.05). Preincubation with sildenafil followed by serotonin enhanced the phosphorylation of ERK1/ERK2 (p-ERK1/ERK2) and further elevated the percentage of cells in S phase and the cell proliferation index compared with that of HT group. While the total ERK1/ERK2 (t-ERK1/ERK2) was not statistically different among these groups. Conclusions Sildenafil potentiates the proliferative effect of serotonin on pulmonary arterial SMCs via upregulating phosphorylation of ERK1/ERK2.

Objective To analyze the detection results of 75 suspected type A influenza cases and to understand the distribution situation of influenza patients to provide the guidance for its early prevention and control .Methods The nasopharyngeal swabs specimens from 75 patients with suspected type A influenza were detected for understanding the viral infection situation by real‐time fluorescent quantitative PCR .Results In the specimens of 75 suspected patients ,13 cases with type A influenza virus were detected out ,in which 9 cases were the H1N1 subtype ,mean age > 50 years old .The temporal distribution was 3 cases in March 2013 ,1 case in April ,7 cases in January 2014 and 2 cases in February .No H7N9 subtype was detected out .Conclusion Real‐time fluores‐cent quantitative PCR has the characteristics of rapidness and high accuracy .The winter is the high onset season of flu ,elderly pa‐tients are susceptible to influenza virus .The monitoring and early treatment should be strengthened .

Objective To study the feasibility of dose verification with Octavius 4D system in volumetric modulated arc therapy (VMAT).Methods Twenty patients were treated with the VMAT.Their treatment plans were performes and then acquired the measured data with the Octavius 4D system.In addition,the treatment plans are used in the CT phantom for Octavius 4D system,and acquire the simulated data.We use the γ pass rate to analysis the measured and simulated results for the conditions:the dose deviations are 2％3％5％,the displacement deviations are 2 mm/3 mm/5 mm and the thresholds are 5％ 10％,and got the Gamma pass rate and fitness of profile curve.Results The distributions of measured and simulated dose are matched well,and the fitness of profile curve are also matched well.The averaged Gamma pass rate is 97.78％ in the case 3 mm 3％ 10％.Conclusions Octavius 4D system can meet the dose verification for VMAT treatment.The measured plane is always following the rotating frame and perpendicular to the beam.

ObjectiveTo investigate and eveluate the significance of serum and urinary L-selectin in patients with transitional cell carcinoma.MethodsSerum and urinary samples were obtained before operation and 6 days after surgery from 46 patients with transitional cell carcinoma of bladder and from 14 patients with benign prostate hyperplasia.The serum and urinary L-selectin levels were measured using ELISA.ResultsThe preoperative serum concentration of L-selectin was significantly higher in patients with invasive bladder cancer [(766.3?135.1)ng/ml] than those with superficial bladder cancer [(563.8?118.2)ng/ml],P

Objective To highlight the diagnose and treatment of stationary paraganglioma in retroperitoneum.Methods Ten cases of stationary medullary chromaffinoma of retroperitoneum proven pathologically from 1997 to 2007 were reviewed and sutdied.Hypertension were not observed in all patients,who were admitted to hospital through routine phisical examination.Endocrine secretion examinations,B-US,CT,MRI,131IMIBG,and octreotide were used to dianose the diease.Among all 10 patients,4 cases took ?-receptor blocker for 2～4 weeks preoperatively.3 were operated laparoscopically,7 were operated with lumber incision.Results Hypertension and urinary catecholamine was usually normal or elevated lightly in stationary medullary chromaffinoma of retroperitoneum.131and octreotide have high sensitivity and accuracy in diagosing medullary chromaffinoma.4 were near renal hilum,6 was in para-aorta.They were from 3 cm to 15 cm in size.The results of pathology were paraganglioma in 7 and malignant paraganglioma in 3.Tumor markers were positive,such as cgA,syn,NSE and s-100.There was 1 case metastasis 1 years later.Conclusion Once the correct diagnosis is made,surgical treatment should be carry out on the basis of correct drug preparation.Intimate follow-up is necessary and important.

Objective To construct TIZ gene RNA interference plasmid,as well as study the biological effects of down-regulating of TIZ gene on ovarian cancer cells.Methods According to the mRNA sequence of TIZ on GenBank,3 pairs of small interference RNA (siRNA) for TIZ gene expression were designed and introduced into SKOV3 cells by liposome transfection reagent; the real-time quantitative PCR (QRT-PCR) was used to detect the interference efficiency and selected the most efficiency siRNA segment;pGPU6/GFP/Neo carrier was used to construct pGPU6/GFP/Neo-siRNA-TIZ-573 restructured interference plasmid,introduced into SKOV3 ccll (sh-TIZ-573/SKOV3).The cell growth curves were made by methyl thiazolyl tetrazolium (MTT) method.The clonogenicity efficiency was observed by clony formation test.The cell cycles were measured by flow cytometry (FCM).The ability of invasion,metastasis and adhesion of ovarian cancer cells were detected by the matrigel invasion assay,transwell migration assay and adhesion assay,respectively.Results QRT-PCR results showed that ovarian cancer SKOV3 cells which transfected by recombinant plasmids pGPU6/GFP/Neo-siRNA-TIZ-573 could down-express TIZ gene.The cell growth curves,FCM and clony formation tests showed that the growth and proliferation of sh-TIZ-573/SKOV3 cells were significantly speeded up compared with the control cells (P ＜ 0.01).There was no significant difference in cell invasion,migration and adhesion between sh-TIZ-573/SKOV3 cells [(48.5 ± 1.7) ％,(53.6 ±3.4)％,(64.9 ±5.0)％,respectively] and the control cells (P ＞0.05).Conclusion The growth of ovarian cancer cells could be speeded up by down-expressing of TIZ gene,which TIZ gene may be play a biological role as a tumor suppressor gene.

Objective To evaluate the clinical characteristics, diagnosis and treatment procedures for adrenal pheochromocytoma and adrenal ganglioneucoma.Methods Thirty-two cases of adrenal pheochromocytoma and 14 cases of adrenal ganglioneucoma surgical treated between 1999—2005 were analysed, The average of ganglioneucoma was 39 years old (16～63). tumor size ranged from 2.5 cm to 15 cm. The mean age of pheochromocytoma was 42 years old (19～74), tumor size ranged from 2cm to 16 cm. Tumor located in the adrenal area. All cases accepted surgical operation and were confirmed by pathology. Results Most cases of adrenal ganglioneucoma did not have any symptom but were founded incidentally. Differentials were obviously between ganglioneucoma and pheochromocytoma. Conclusion Both adrenal ganglioneucoma and pheochromocytoma are rare diseases.CT、urinary CA and 131I-MIBG are good methods for diagnosis.The final diagnosis depends on the pathology, surgical resection of tumor is a preferred choice.

[Objective ] Intrastriatal Parr interneurons are considered to be the junction between striatal input and output neurons. This study purposes to confirm the morphology of Parr neurons and their distribution in striatum. [Method] Adult male SD rats were perfused and the brains were removed. The sections were conducted with a semiconductor-frozen microtome, then single labeling and double labeling were immunocytochemically conducted with PAP technique. Positive neurons were observed, counted, and calculated, and analyzed with SPSS. [Results] The distribution of Parr positive neurons in striatum was inhomogeueous, and was the most dense in the dorsal lateral area (P<0.001) ; the distribution of Parr positive neurons in matrix compartment was apparently more than patch compartment (82.0% vs 18.0%, P<0.001); Parr positive neurons in striaturn were medium-sized cell bodies with polygonal or oval in shape (mean diameter is 11.68 μm), and were larger in the lateral area than that in the medial area (P<0.01) ; the positive dendrites were dense and smooth without dendrite spines; the positive axons 1 were slender and their collaterals extended in striatum. [Conclusion] The characteristic of Parr positive neurons collecting mostly in lateral striatum and matrix eompartment, as well as their traits of intemeurons, indicated that they would affect the striatal projection neuronal function.

Hepcidin are small cationic peptides with antibacterial activity expressed mainly in the liver of living organisms, and they play important roles in the host's immune response against microbial invasion and regulation of iron metabolism. Thus, they are considered to be good substitutes for traditional antibiotics. It is a good choice that the antimicrobial peptides are prepared by recombinant DNA expression. In the present study, two hepcidin mature peptide cDNAs from channel catfish (Ictalurus punctatus) (mCH) and tilapia (Oreochromis niloticus) (mTH) were connected by SOE-PCR in order to obtain more recombinant hepcidin with broad antimicrobial spectrum, and EcoR I and Not I sites were added to 5'- and 3'- ends of the fragment, respectively. The recombinant eukaryotic expression vector "pPIC9K-mCH-mTH" was successfully constructed, and transformed into Pichia pastoris GS115. The transformants containing multicopy gene insertion were selected by using different concentrations of G418 and other specific mediums, and identified by PCR for yeast genomic DNA. Expression was induced by adding 1% methanol at 30 degrees C for different times. Tricine-SDS-PAGE analysis demonstrated that the most appropriate expression time was 72 h, at which a high expression yield (77 mg/L) for the target protein was exhibited. The highly purified target protein was obtained from the fermentation supernatant by SP-Sepharose cation exchange chromatography. Bacteriostatic activity assay demonstrated that the fermentation supernatant containing the target protein and purified recombinant target protein had bacteriostatic activities against gram-positive and gram-negative bacterium. The present result provides the important initial value for industrial production of hepcidin antimicrobial peptide.
Animals ; Electrophoresis, Polyacrylamide Gel ; Fish Proteins ; biosynthesis ; genetics ; Fishes ; genetics ; Gram-Negative Bacteria ; Gram-Positive Bacteria ; Hepcidins ; biosynthesis ; genetics ; Pichia ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis