AIM: To investigate the effect of selective silencing of SLC7a8 on uptaking L-dopa in renal tubular epithelial cells of rat (NRK-52E). METHODS: The three siRNAs targeting SLC7a8 (siRNA-1, siRNA-2, siRNA-3) were designed and synthesized. A siRNA with nonspecific coding sequence (siRNA-con) was used for control. All siRNAs were transfected into NRK-52E cells. The siRNA-con transfected group, blank control group and gene-specific silencing SLC7a8 group were set up. The efficiency of transfection was estimated by flow cytometry. The efficiency of RNA interference was detected and screened by RT-PCR preliminarily, and was followed by Western blotting at protein level. The concentrations of L-dopa uptake into the NRK-52E cells were detected by ultraviolet spectrophotometry at different time points (6, 12, 24, 36, 48, 60, 72 and 120 min). RESULTS: The transfection efficiency was 94% detected by flow cytometry. The initial screening of RT-PCR showed that the efficiencies of RNA interference of siRNA-1 and siRNA-3 were higher, and siRNA-3 was the highest at protein level determined by Western blotting. No distinctive change was found between siRNA-con treated NRK-52E and blank control cells. The L-dopa uptake at different time points (6, 12, 24, 36, 48, 60, 72 and 120min) in siRNA-interference group was lower than that in siRNA-con transfected group and blank control group. No significant difference of L-dopa uptake between siRNA-con group and blank control group was observed. CONCLUSION: RNA interference technology selectively down-regulates SLC7a8 expression in rat renal tubular epithelial cells. The L-dopa uptake is also decreased after specifically silencing the slc7a8 expression.

Background Many ophthalmologists have proved that the intravitreal injection of plasmin can safely induce posterior vitreous detachment(PVD),but if it can generate the complete PVD need further to seek confirmation.Researches showed that the safe dose and toxicity dose of dispase are very near,so its application is limited.Whether hyaluronidase can induce PVD is still in controversy.Objective This study is to clarity the mechanism of pharmacological vitreolysis with plasmin and hyaluronidase.Methods Plasmin 4μmol/L,2μmol/L and 1μmol/L,plasmin 1μmol/L+ hyaluronidase 20μmol/L,hyaluronidase alone were intravitreally injected in lateral eye of 4 clean New Zealand white rabbits respectively,and 0.1mL BSS was injected as control group.Electron immunocytochemical technique was used to detect the laminin and fibronectin of interface between vitreous and retina in 7 days after intravitreal injection.Other 14 eyes of 7 clean New Zealand white rabbits were used in this study.Plasmin 1μmol/L + hyaluronidase 20μmol/L was intravitreally injected in the lateral eyes,and only plasmin 1μmol/L was injected in the fellow eyes.Plasmin activity in vitreous was evaluated in 15 and 30 minutes,1 hour,2,3,6,12 hours after intravitreal injection.The use of animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The amounts of laminin and fibronectin in the vitreoretinal interface were decreased in 4μmol/L plasmin group,2μmol/L plasmin group,1μmol/L plasmin group,1μmol/L plasmin+20μmol/L hyaluronidase group compared with control group(P＜0.01).No significant difference was seen in the density of gold particles of anti FN between 20μmol/L hyaluronidase group and control group (P＞0.05).The change of amounts of fibronectin in the vitreoretinal interface was similar to that of laminin.Plasmin activity remained the highest level 1 hour after injection and thereafter gradually decreased and extincted in 12 hours and presented the same trend between plasmin 1μmol/L+hyaluronidase 20μmol/L group and only plasmin 1μmol/L group.Conclusion The mechanism of pharmacological vitreolysis is to dissolve laminin and fibronectin in the interface between vitreous and retina and therefore induce PVD.Combination of plasmin with hyaluronidase can increase the efficiency of pharmacological vitreolysis.The optimum selection of drug in inducing PVD should consider not only its role of lysis laminin and fibronectin but also the role of liquefying the vitreous.

OBJECTIVE:To prepare Activated carbon N-acetylcysteine microcapsule (ACNAC),and to optimize preparation technology. METHODS:ACNAC was prepared by emulsion cross-linked method using biodegradable material gelatin as capsule wall material. Using comprehensive evaluation index of drug-loading amount,entrapment rate and particle size distribution percent-age(the percentage of 80-140 μm particle)as index,drug-loading ratio,amount of gelatin,mixing speed and the amount of emul-sifier as factors,single factor test and orthogonal test were used to optimize formulation technology. The technology was validated and distribution of particle size of ACNAC was determined. RESULTS:The optimal formulation technology was as follows as drug-loading ratio 1∶1,gelatin 15%,emulsifier 2.0%,mixing speed 1 000 r/min. Average drug-loading amount of 6 batches of ACNAC was 15.9%(RSD=1.21%),average encapsulation efficiency was 78.1%(RSD=1.11%)and average particle size distri-bution percentage was 81.9%. CONCLUSIONS:ACNAC is prepared successfully,and formulation technology is reasonable and feasible.

OBJECTIVETo observe the clinical efficacy of Tongfu Mixture (TM) for post-ERCP pancreatitis (PEP).

METHODSTotally 54 PEP patients were randomly assigned to the control group (treated by routine therapy, 26 cases) and the TM treatment group (treated by TM, 28 cases). Clinical indices including the alleviation time of abdominal pain/distention, gastrointestinal function recovery time, and the post-surgical length of stay were observed. Blood amylase (AMY), C-reactive protein (CRP), plasma endotoxin (PLS), TNF-alpha, and IL-6 were detected before surgery, 12 h, 48 h, and 96 h after surgery.

RESULTSThe alleviation time of abdominal pain/distention, the gastrointestinal function recovery time, and the post-surgical length of stay were obviously shorter in the TM treatment group than those in the control group (P < 0.05). The recovery of AMY and CRP were better in the TM treatment group than in the control group at post-operative 48 h and 96 h (P < 0.05). The levels of LPS, TNF-alpha, and IL-6 were lower in the TM group than in the control group at post-operative 96 h (P < 0.05).

CONCLUSIONTM showed better clinical efficacy and could significantly decrease the post-surgical length of stay. post-ERCP pancreatitis; integrative medicine; Tongfu Mixture

Adult ; Cholangiopancreatography, Endoscopic Retrograde ; adverse effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; drug therapy ; etiology ; Phytotherapy

Animals ; Apolipoproteins E ; deficiency ; genetics ; Blood Vessels ; anatomy & histology ; Fluorescence ; Lipofuscin ; Mice ; Mice, Knockout

Objective (1) To evaluate the effect of insertion of two 15-amino-4,7,10,13-tetraoxapentadecanoic (2 PEG4 ) linkers into cyclic Arg-Gly-Asp (RGD) Dimer E [c(RGDfK)]2 on receptor binding in vitro, (2) to assess its biodistribution in vivo and (3) to investigate the value of 99Tcm labeled 2PEG4-Dimer for integrin αvβ3-positive tumors imaging.Methods The expression of U87 human glioma cells and integrin αv β3 was determined by immunofluorescence staining.The half-inhibition concentrations (IC50) for 125 I-cyclo (Arg-Gly-Asp-D-Tyr-Lys) (c(RGDyK) ) of c ( RGDyK ), hydrazinonictinamide ( HYNIC )-Dimer and HYNIC-2PEG4-Dimer binding to integrin αvβ3 were measured.99Tcm-HYNIC-Dimer and 99Tcm-HYNIC-2PEG4-Dimer were synthesized using non-SnCl2 formulation.Biodistribution and imaging studies were performed in nude mice bearing human glioma xenografts.The unpaired t test was used for statistical analysis.Results The labeling yield of the two radiotracers was more than 95%, and the radiochemical purity was more than 99% through Sep-Pek C18 cartridge.HYNIC-2PEG4-Dimer had significantly higher binding affinity of integrin αvβ3 than c(RGDyK) and HYNIC-Dimer (IC50 = 0.8 nmol/L, 27 nmol/L and 2.4 nmol/L, respectively).Biodistribution study showed that 99Tcm-HYNIC-2PEG4-Dimer was mainly excreted via the kidney.The tumor uptake of 99Tcm-HYNIC-2PEG4-Dimer was higher than that of 99Tcm-HYNIC-Dimer at 2h post injection ((5.71 ±0.96) and (2.10 ±0.50) % ID/g, t =4.80, P＜0.05).The xenografted tumors were visible at 0.5 h post injection and the image contrast increased with time due to the tracer clearance of the background tissue.Conclusion 99 Tcm-HYNIC-2PEG4-Dimer is a promising radiotracer for integrin αvβ3-positive tumor imaging.

This paper introduces the old nine-needle including its name, shape, function, manipulation and origination, as well as systematically introduces the new nine-needle as a new product which has been developed under the guidance of modern science and technology based on the old nine-needle since 1949. As a new tool, the new nine-needle provide much more choice for acupuncture treatment and obtained satisfied clinical effect. During the development of research and clinical practice on the new nine-needle, its theory has been consummated successively. Combining various kinds of needles for treatment is not only paying attention to the specificity of one kind of needles, but also emphasizing the whole function of all kinds of needles, which makes the new nine-needle formed the characteristic and distinct theory of its own.
Acupuncture Therapy ; history ; instrumentation ; methods ; China ; History, Ancient ; Needles

Objective To investigate the effect of telmisartan on the expression of peroxisome proliferator activated receptor gamma(PPARγ)and its regulation in myocardial remodeling in spontaneously type 2 diabetic male Otsuka Long-Evans Tokushima Fatty(OLETF)rats fed with high-fat diet.Methods Twenty-eight four-week-old male OLETF rats were fed with high-fat diet. From 22-week of age, the pre-diabetic OLETF rats were randomly assigned to three groups:telmisartan-treated group[O-T group,5 mg/(kg·d),n=10],pioglitazone-treated group[O-P group,10 mg/(kg·d),n=8]and untreated group ( O-C group, equal volume of normal saline, n=10), continuously administration for 22-week. Twelve sex and age matched Long-Evans Tokushima Otsuka(LETO)rats were used as control(LETO group).At 22 and 48-week of age, the glucose tolerance of the rats was assessed by the oral glucose tolerance test(OGTT).At 48 weeks of age,five rats were randomly selected from each group,and clamp experiments were carried out.The glucose infusion rate from 60 min to 120 min(GIR60-120)was measured.All rats were sacrificed and the myocardial tissues were dissected.The ratio of heart weight to body weight(HW/BW)was calculated.Blood samples were collected,and serum PPARγ,tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6)and adiponectin were measured using ELISA and radioimmunoassay.The mRNA expressions of IL-6,PPARγ1 and PPARγ2 were measured by real-time PCR.The protein expression levels of PPARγ,adiponectin,IL-6 and NF-κB were determined by Western blot assay.The myocardial pathological changes were observed under light microscope (HE staining, Masson staining and PAS staining), and ultrastructural changes were observed under transmission electron microscope.Results At 22-week of age,neither type 2 diabetes mellitus(T2DM)nor IGT were found in three groups.At 48-week of age,T2DM was found in seven rats of O-C group,and IGT was found in two rats.T2DM was found in one rat of O-C group,and IGT was found in three rats.Neither T2DM nor IGT was found in the other two groups.GIR60-120and HW/BW were all significantly lower in the O-P group and O-T group than those of the O-C group at 48-week of age (P<0.05). Systolic blood pressure(SBP)and diastolic blood pressure(DBP)were significantly lower in O-T group than those in other three groups(P<0.05).Compared with the O-C group,the serum levels of PPARγ and adiponectin were significantly up-regulated,whereas the serum levels of IL-6 and TNF-α were down-regulated by telmisartan administration in O-T group (P<0.05).There were no significant differences in the above indexes between O-P and O-T groups.The results of real-time PCR and Westen blot assay showed that the mRNA expression of PPARγ1 was increased,and IL-6 expression decreased in O-T group compared with those of O-C group. The protein expressions of PPARγ and adiponectin were increased, and protein expressions of NF-κB and IL-6 were significantly decreased in O-P group and O-T group compared with those of O-C group(P<0.05).In the O-C group,the arrangernent of myocardial cells was irregular,myocardial fibers were swollen,a large amount of fibrotic tissue in the myocardial interstitium, and glycogen accumulation under light and electron microscope. Besides, myofibril breakage and perinuclear space expansion, myocardial mitochondria were apparently damaged or even dissolved. Compared with the O-C group, myocardial fibers arranged neatly, no obvious glycogen deposition and the ultrastructural changes of myocardium were obviously reduced in O-T group and O-P group. Conclusion Telmisartan can increase the expression level of PPARγ in the serum and myocardial tissue, reduce myocardial fibrosis and alleviate cardiac remodeling in the high-fat-diet OLETF rats.

Objective To study the effects of coal burning related endemic fluorosis on body development and intelligence levels of fluorosic children in Zhijin county.Methods One hundred and seventy-six fluorosic children and 50 healthy(without fluorostic teeth) children aging 7-12 years who were sampled in cluster sampling were examined for urine fluorosis,physical examination and intelligence tests in Zhijin county,with coal burning pollution related fluorosis.Results Physical development of sick children and the children in control group were in low levels. The intelligence levels and the leves of urine fluorosis in study group were lower than those in control group and there was a significant difference(P

OBJECTIVETo explore the mechanism of Shenqi compound recipe (SQCR) anti-earlier diabetic artherosclerosis in GK rats.

METHODFour-month specefic pathogen free (SPF) GK rats were divided randomly according to blood glucose level into four groups: model group (5 mL x kg(-1) x d(-1) sterile water), ramipril group (positive control, 1 mg x kg(-1) x d(-1)), SQCR low dosage (0.72 g x kg(-1) x d(-1)) and SQCR high dosage group (2.88 g x kg(-1) x d(-1)) and Wistar rats as normal control group(5 mL x kg(-1) x d(-1) sterile water). GK rats took high-fat diet freely and meanwile were injected N-omega-nitro-L-arginine methyl ester (L-N-AME) intra-peritoneally with the dose of 10 mg x kg(-1) x d(-1) in order to induce earlier diabetic artherosclerosis, while normal control group took regular diet and were injected normal saline intra-peritoneally. In the experiment periods, each group was administrated correspondent substance respectively for 32 d. At the end, sampling blood by abdominal aorta and picking aorta on ice. Determined monocyte chemoattractant protein-1 (MCP-1) concentration by ELISA, messenger ribonucleic acid (mRNA) expression of MCP-1 and peroxisome proliferator-activated receptor gamma (PPARgamma) in aorta by reverse transcriptase PCR (RT-PCR).

RESULTConcentrations of MCP-1 in serum in SQCR low and high dosage groups and the mRNA expression of MCP-1 in SQCR high dosage group were all decreased significantly compared with model group (P < 0.05). The mRNA expression of PPARgamma in SQCR low and high dosage groups all increased compared with model group (P < 0.05 or P < 0.01).

CONCLUSIONInhibiting the mRNA and protein expression of MCP-1 and upregulating the mRNA expression of PPARgamma in aorta might be contribute to SQCR anti-earlier diabetic artherosclerosis in GK rats partly.

Animals ; Aorta ; metabolism ; Astragalus membranaceus ; chemistry ; Atherosclerosis ; etiology ; metabolism ; Chemokine CCL2 ; biosynthesis ; blood ; genetics ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Male ; PPAR gamma ; biosynthesis ; genetics ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar