AIMTo elucidate possible mechanisms underlying the differences between 1S-[1a,2b,3b,4a(S*)]-4-[7-[[1-(3-chloro-2-thienyl)methylpropyl]propyl-amino]-3H-imidazo[4,5-b]pyridyl-3-yl]-N-ethyl-2,3-dihydroxycyclopentane carboxamide (AMP- 579) and adenosine in pharmacological and clinical effects. METHODSNa+/Ca2+ exchange current was recorded by patch-clamp technique in whole-cell configuration. RESULTSAMP579 significantly enhanced both outward and inward Na+/Ca2+ exchange currents in a concentration dependent manner. Neither infusion of an adenosine A1 receptor antagonist PD116948 30 μmol*L-1 or an adenosine A2 receptor antagonist DMPX 10 μmol*L-1 nor a protein kinase A special blocker KT 5720 0.2 μmol*L-1 or a protein kinase C special blocker GF 109203X 0.4 μmol*L-1 had effect on Na+/Ca2+exchange current increased by AMP579, suggesting that AMP579 possess a direct activating effect on Na+/Ca2+ exchange current. CONCLUSIONAMP579 possibly possesses a direct activating effect on Na+/Ca2+ exchange current.

Objective To investigate the relationship of RhoA and Snail expressions, and the invasion and metastasis in salivary adenoid cystic carcinoma (SACC). Methods The expressions of RhoA protein and Snail protein in 55 samples of SACC (SACC group ) and 20 samples of para-carcinoma normal tissues(control group) were detected using immunohisto?chemical method. The relationship between RhoA protein and Snail protein expressions and clinical and pathological charac?teristics were analyzed. Results The positive expressions of RhoA protein (69.1% vs 5.0%) and Snail protein (72.7% vs 10.0%) were significantly higher in SACC group than those in control group (P < 0.05). The positive expression rates of RhoA protein and Snail protein were significantly higher in patients with lymph node metastasis than those in patients with?out lymph node metastasis. The positive expression rates of RhoA protein and Snail protein were significantly higher in pa?tients atⅢ+Ⅳstage than those in patients atⅠ+Ⅱstage. The positive expression rates of RhoA protein and Snail protein were significantly higher in substantive carcinal tissues than those in screen roller type and tubular carcinal tissues. The posi?tive expression of Snail protein was significantly higher in substantive and tubular carcinal tissues than that in screen roller type carcinal tissues (P<0.05). There were no significant differences in positive expression rates of RhoA and Snail between different gender, age and different carcinal tissues. There was a positive correlation beween expression rates of RhoA and Snail protein in SACC (r=0.414, P<0.001). Conclusion RhoA and Snail may both facilitate the infiltration and metastasis of SACC through RhoA/ROCK/PKD1/NF-kappa B/Snail signaling pathways.

Objective To evaluate of the clinical efficacy of two kinds of methods extracorporeal shock wave lithotripsy in treatment of ureteral calculi. Methods A retrospective analysis was conducted on the data of 108 patients with acute renal colic and ureteral stones (calculi diameter<2 cm) from March 2011 to April 2013. The patients were divided into two groups. 69 cases in A group underwent emergency positioning urography ureteral catheterization combined extracorporeal shock wave lithotripsy, 39 cases in B group underwent emergency extracorporeal shock wave lithotripsy. Results A group:lithotripsy was successful in 63 cases among 69 cases at the first time,the first success rate was 91.33% (63/69), the second success rate was 94.2%(65/69),2 cases of retrograde ureteroscopy failed, 2 cases of self-discharge after intubation stones, the recurrent renal colic rate was 14.3%(9/63),the use of analgesic drugs rate was 11.1%(7/63) . B group:the first success rate of stone clearance rate was 74.4%(25/39),the second success rate of stone clearance rate was 87.2%(34/39),5 cases underwent ureteroscopy lithotripsy, the renal colic recurrence rate was 20.5% (8/39), the use rate of analgesic drugs was 15.4% (6/39) . Conclusion Extracorporeal shock wave lithotripsy assisted by urography through retrograde ureteral catheterization has satisfying therapeutic effect in treatment of patients with ureteral calculi.

Objective To study the effects of IL-4 in prostate cancer cells glycometabolism and proliferation. Methods We used IL-4 to treat PC3 cells, then tested the changes of LDH-A expression by RT-qPCR, Western Blot, CCK-8 and lactate production assay. Results Our data showed that IL-4 induced LDH-A up-expression in PC3 cells at mRNA and protein levels. Also, IL-4 promoted the proliferation activation and increased lactate production in PC3 cells. Conclusion IL-4 can strengthen the proliferation activation in PC3 cells by up-regulating LDH-A expression.

Aim To examine the effect of zacopride,a specific inward rectifier potassium channel(IK1)agonist,on L-thyroxine(T4)-induced ventricular remodeling and the underlying mechanism.Methods SD rats were randomly divided as control,L-thyroxine(L-thy,1 mg·kg-1·d-1,ig,10 d)model,L-thy +zacopride(5,15,50 μg·kg-1,respectively,ip),L-thy+zacopride(15 μg·kg-1)+chloroquine(7.5 μg·kg-1,ip)and L-thy+captopril(100 mg·kg-1·d-1,drinking water)groups.Echocardiography and cardiac hypertrophic indexes were measured to confirm the establishment of the ventricular remodeling model.The changes of IK1 and L-calcium current(ICa-L)were detected by whole cell patch clamp technique.The confocal microscopy and fluorescent indicator Fluo-4 were applied to examine the intracellular Ca2+ concentration([Ca2+]i)of isolated adult rat ventricular myocytes.Results L-thyroxine induced left ventricular hypertrophy with increased ratio of heart weight(HW)to body weight(HW·BW-1),ratio of left ventrical weight(LVW)to body weight(LVW·BW-1),left ventricular dimension in diastole(LVIDd),left ventricular dimension in systole(LVIDs),interventricular septum thickness(IVS)and decreased ejection fraction(EF),fractional shortening(FS)(P<0.01).Patch clamp data suggested IK1 was downregulated,while ICa-L was upregulated(P<0.01).In isolated adult cardiomyocytes,L-thyroxine increased the cell area and [Ca2+]i(P<0.01).Zacopride treatment obviously alleviated cardiac remodeling,improved cardiac function,reversed the changes of IK1 and ICa-L,and significantly attenuated intracellular calcium overload(P<0.01).The optimum dose of zacopride in vivo was 15 μg·kg-1 at which the effect was compared favourably with captopril,a classical anti-remodeling agent.Low-dose IK1 atagonist chloroquine could reverse the effect of zacopride(P<0.01).Conclusion Via activating IK1,zacopride could significantly decrease Ca2+ influx and intracellular calcium overload thereby inhibiting L-thyroxine-induced cardiac ventricular remodeling.

OBJECTIVETo reveal the variation content of polysaccharides in cultivated Dendrobium candidum and the relationship between germplasms, harvesting and polysaccharides content for the breeding of quality of D. candidium.

METHODThe morphological characteristics were recorded when 33 samples were collected. The content of polysaccharides was determined by phenol-sulphuric acid method.

RESULTThe average content of polysaccharides in 2-year-old samples was 34.47% (25.63%-41.65%). The polysaccharides content of samples were significantly different among germplasms and physiological ages.

CONCLUSIONThe polysaccharides content of cultivated D. candidum is higher than that of wild materials. Germplasms and physiological age impact on the polysaccharides content significantly. D. candidum breeding and the control of harvesting can increase polysaccharides content.

Breeding ; Dendrobium ; chemistry ; genetics ; growth & development ; Drugs, Chinese Herbal ; analysis ; Polysaccharides ; analysis

Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.