Objective To construct transgenic mice tissue-specifically co-expressing human DAF/CD59 in the vascular endothelia and to investigate the ability to protect against human comple- ment-mediated attack.Methods Transgenic mice were generated by co-microinjection of hDAF/CD59 expression constructs driven by the human intercellular adhesion molecule-2(ICAM-2)promoter.PCR and Southern blot of genomic DNA were used to assess the presence of hDAF/CD59 in the genome of the founders,and the expression at protein level was measured by flow cytometry.Immunohistochemis- try was used to detect the distribution of hDAF/CD59.An ex vivo perfusion model was used to com- pare hearts from these hDAF/CD59 transgenic mice with hDAF hearts.Results After microinjection of genes,11 of 135 mice born were shown to be double-transgenic,and human DAF/CD59 were ex- pressed on the surface of leucocytes in 6 of the 11 DAF/CD59-integrated mice.Expression levels of 6 founders ranged from 80% to 95% of that in human leucocytes.Human DAF/CD59 were strongly expressed in the vascular endothelia of heart,kidney,with little or no positive staining observed in non- endothelial cells.Compared to hDAF hearts that maintained approximately 20% maximum work during perfusion with 20% human plasma,these endothelial-specific hDAF/CD59 hearts were further protected with work maintained at 40% of the maximum level during 60 min.Conclusion The intro- duced hDAF/CD59 genes have been integrated and specifically expressed in the vascular endothelia. The endothelial co-expression of hDAF/CD59 can provide greater protection against human complement-mediated attack than the expression of hDAF alone.

Objective To investigate the proliferation of rabbit adipose-derived stem cell(ADSCs) transfected by adenoviral vector mediated hTGF-?_1 gene and its chondrocyte differentiation potential.Methods The Ad-hTGF-?_1 plasmid vetor which had the hTGF-?_1 gene was developed and transfected the ADSCs.The experimental group was the hTGF-?_1 transfected group.The cells enclosed by alginate were cultured in com- plete chondrogenie medium(CMM).The morphology of the cells were observed,and RT-PCR was used to measure hTGF-?_1 and collagenⅡexpression,at the same time western blot and immunohistochemistry were applied to detect the expression of collagenⅡin ADSCs before and after transfected with hTGF-?_1 gene. Results The hTGF-?_1 transfected ADSCs became the polygon and it proliferated well.The RT-PCR result of hTGF-?_1 on the transfected group was better than the control after transtected for 7 day and 21 day.The dif- ference between the two groups was significant(P

AIMTo determine clonidine in rabbit plasma by LC-MS.

METHODSThe LC-MS system consisted of Waters Alliance 2790 HPLC and Micromass ZQ-4000 MS. The HPLC was performed by using XTerra C18 (150 mm x 2.1 mm ID, 5 microm). The mobile phase, consisting of acetonitrile/ammonium hydrogen carbonate solution, was maintained to a flow-rate of 0.2 mL x min(-1) and the linear gradient elution was adopted. Mass spectrum was obtained by using electrospray ionization interface and the m/z of SIM was 230.

RESULTSThe average recovery was high and the method was reproducible. The calibration curve showed good linearity in the range of 1 - 80 microg x L(-1), the lowest limit of detection was 0.05 microg x L(-1). The Cmax, AUC0-t, and Tmax value of the pharmacokinetics parameter were (27 +/- 9) microg x L(-1), (5,352 +/- 1,121) microg x L(-1), (79 +/- 17) h.

CONCLUSIONThe results demonstrated that the method had high sensitivity, good selectivity, accuracy and precision. It is used to determine the clonidine concentration in plasma. The transdermal patch can deliver clonidine to the surface of rabbit skin stably for periods of up to 1 week after a single application.

Administration, Cutaneous ; Animals ; Antihypertensive Agents ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Clonidine ; administration & dosage ; blood ; pharmacokinetics ; Rabbits ; Spectrometry, Mass, Electrospray Ionization ; methods

Objective To survey and analyze characteristics of brucellosis epidemic in Weinan city of Shaanxi province for the purpose of setting up prevention and control measures for the disease. Methods According to "The Executing Plan for the Work of Surveying Brucellosis Disease in Shaanxi Province", 35 villages(towns) of designated monitoring locations and 24 villages (towns) of randomized monitoring locations in five countries of Weinan were chosen to survey brucellosis disease. The five countries were Chengcheng, Dali, Heyang, Tongguan and Hancheng. High risk populations with a history of contacting livestock and livestock products aged between 7 and 60 underwent clinical and serology examination[rose bengal plate agglutination test(RBPT) and standard tube agglutination test(SAT)]. All manipulation methods and judging standards were in accord with the "Diagnostic Standard for Brucellosis" (WS 269-2007). Results In the designated monitoring location, a total of 8664 people at high risk were investigated, among whom 1407 people were tested by RBPT test and 27 people were positive,the positive rate was 1.92%(27/1407); 27 people were tested by SAT test and 27 people were positive, the positive rate was 100% (27/27); 25 people were diagnosed and the diagnosis rate was 92.59%(25/27). In the randomized monitoring location, a total of 3464 people at high risk were investigated, among whom 411 people were tested by RBPT test and 3 people were positive, the positive rate was 0.73%(3/411 ), 3 people were tested by SAT test which were all positive and made a definite diagnosis. Twenty-eight new cases were made a definite diagnosis and its incidence was 2.06 in a hundred thousand(28/1 361 618). Conclusions The infection of human brucellosis in Weinan city stays at higher level. The governments should increase input for the monitoring,investigating and disinfecting to prevent the disease from increasing and outspreading.

OBJECTIVE: To study nomal valuels of PI, RI and S/D in healthy male penile dorsum artery (DA) and cavernosal artery (CA). METHODS: 257 healthy mature men were divided into 5 groups by age. Group 1: <30, n=65; Group 2: 30-39, n=83; Group 3: 40-49, n=61; Group 4: 50-59, n=38; Group 5: > or = 60, n=10. Hibateral PI, RI and S/D values of penile dorsum artery and cavernosal artery were examined by Logidop(r)2 Type Digital Doppler Ultrasonography. RESULTS: There were no significant difference for PI, RI and S/D of penile dorsum artery and cavernosal artery not only in personal left and right artery but also in different age groups. Normal values advised: (1)LDA:PI 1.43-3.43, RI 0.72-0.92, S/D 2.68-10.56. (2)RDA PI 1.47-3.47, RI 0.73-0.93, S/D 3.27-10.09. (3)LCA:PI 1.49-3.21, RI 0.74-0.90, S/D 3.17-9.55. (4)RCA:PI 1.93-3.27, RI 0.72-0.90, S/D 3.22-9.42. CONCLUSION Doppler ultrasonography is a favorable method in filtering penile arterial function.
Adult ; Age Factors ; Arteries/diagnostic imaging* ; Blood Flow Velocity ; Blood Pressure ; Humans ; Impotence, Vasculogenic/diagnosis* ; Male ; Middle Aged ; Penis/diagnostic imaging* ; Ultrasonography, Doppler

The present study aimed to investigate the protective effect and mechanism of hydrogen sulfide donor NaHS administration against gastric mucosal injury induced by gastric ischemia-reperfusion (GI-R) in rats. GI-R injury was induced by clamping the celiac artery of adult male SD rats for 30 min and followed by reperfusion for 1 h. The rats were randomly divided into sham group, GI-R group, NaHS group, glibenclamide group and pinacidil group. Gastric mucosal damage was analyzed with macroscopic injured area, deep damage was assessed with histopathology scores, and the hydrogen sulfide concentration in plasma was determined by colorimetric method. The results showed that pretreatment of NaHS significantly reduced the injured area and deep damage of the gastric mucosa induced by GI-R. However, NaHS did not significantly alter the levels of hydrogen sulfide in plasma 14 d after NaHS administration. The gastric protective effect of NaHS during reperfusion could be attenuated by glibenclamide, an ATP-sensitive potassium channel (K(ATP)) blocker. However, K(ATP) opener pinacidil inhibited the GI-R-induced injury. These results suggest that exogenous hydrogen sulfide plays a protective role against GI-R injury in rats possibly through modulation of K(ATP) channel opening.
Animals ; Gastric Mucosa ; pathology ; Hydrogen Sulfide ; metabolism ; Ischemic Preconditioning ; methods ; KATP Channels ; metabolism ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Stomach ; blood supply ; Sulfides ; pharmacology

OBJECTIVETo investigate the regional spread of micrometastatic nodules in the mesorectum from low rectal cancer, and provide further pathological evidence to optimize radical resection procedure for rectal cancer.

METHODSA total of 62 patients with low rectal cancer underwent low anterior resection and total mesorectal excision (TME) was included in this study. Surgical specimens were sliced transversely and serial embedded blocks were made at 2.5 mm interval, and paraffin sections were stained with hematoxylin and eosin. The mesorectum on whole-mount sections was divided into three regions: outer region of mesorectum (ORM), middle region of mesorectum (MRM) and inner region of mesorectum (IRM). Microscopic spread were examined microscopically on the sections for the distribution in different mesorectal regions, frequency, types, involvement of lymphatic system and correlation with the primary tumor.

RESULTSMicroscopic spread of the tumor in mesorectum and ORM was observed in 38.7% (24/62) and 25.8% (16/62) of the patients, respectively. Circumferential resection margin (CRM) involved by microscopic tumor foci occurred in 6.5% (4/62) of the patients, and distal mesorectum (DMR) involvement was recorded in 6.5% (4/62) with a spread extent within 3 cm of distal border of the main lesions. Most (20/24) of the patients with microscopic spread in mesorectum were in TNM stage III.

CONCLUSIONResults of the present study support that complete excision of mesorectum without destruction of the ORM is essential for surgical management of low rectal cancer, and an optimal DMR clearance resection margin should not be less than 4 cm.

Adenocarcinoma ; pathology ; surgery ; Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Mesentery ; pathology ; surgery ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Neoplasm, Residual ; Neoplastic Cells, Circulating ; pathology ; Peritoneal Neoplasms ; pathology ; surgery ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; surgery

Cell Line ; Dialysis Solutions ; adverse effects ; Epithelial Cells ; metabolism ; Fibrosis ; Humans ; Peritoneal Dialysis, Continuous Ambulatory ; adverse effects ; Peritoneum ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; analysis ; RNA, Small Interfering ; pharmacology ; Transforming Growth Factor beta ; antagonists & inhibitors ; genetics ; Transforming Growth Factor beta1

OBJECTIVETo study the effects of methyl jasmonate (MJ) on the accumulation and release of tanshinones in suspension cultures of Salvia miltiorrhiza hairy roots.

METHODAfter 18 day's suspension culture of S. miltiorrhiza hairy roots induced by Agrobacterium rhizogenes ATCC15834, the chemical elicitor--methyl jasmonat was added into 6-7V suspension cultures and at the same time, tanshinones contents (including cryptotanshinone and tanshinone II(A)) on the day 2, 6 and 9, after dealing with MJ, was quantified by HPLC.

RESULTAfter dealing with MJ on the day 2, 6 and 9, the concentration of cryptotanshinone reached to 0.039, 0.204, 0.571 mg x g(-1) respectively,and tanshinone II(A) reached 0.251, 0.601 and 1.563 mg x g(-1) respectively. After 9 day's treatment by MJ, the maximum increase of cryptotanshinone and tanshinone II(A) were 23.8 fold and 6.2 fold higher than that of the control respectively.

CONCLUSIONMJ could stimulate the accumulation of tanshinones in S. miltiorrhiza and have released them into the culture medium.

Acetates ; pharmacology ; Culture Techniques ; Cyclopentanes ; pharmacology ; Diterpenes, Abietane ; Oxylipins ; pharmacology ; Phenanthrenes ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; metabolism ; Plants, Medicinal ; drug effects ; growth & development ; metabolism ; Salvia miltiorrhiza ; drug effects ; growth & development ; metabolism

OBJECTIVEThe multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity.

METHODDuringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations.

RESULTThe results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group.

CONCLUSIONThis method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.

Angelica ; chemistry ; Chemical Phenomena ; Coumarins ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; methods ; Molecular Structure