Objectives: To improve the procedures for establishment of focal cerebral ischemic rat models with the intraluminal suture method and to elevate the survival rate of the models. Methods: Sixty male SD rats (weight, 250-270 g) were randomly allocated into 3 groups (n =20 in each group). Control group used the conventional surgical method, the pterygopalatine arteries were exposed and ligated during the procedure, and the rats had free access to food after the procedure; gavage group used the same method as the control group, but postoperative gavage was performed; protective group did not ligate the pterygopalatine arteries, but the carotid arteries and vagus nerves were carefully separated. Trying to avoid stretching the vagus nerves during the procedure and postoperative gavage was performed. The body weight of the rats in each group was weighed continuously before and after modeling, and the neurological deficit scores were measured; the survival rate of rats in each group was calculated at day 3, 14, and 28, respectively after modeling; the brain infarct volume in each group was calculated at day 28 after modeling. Results: Circled digit oneThe mean time of model production was 19.5 min in the protective group, and it was shorter than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit twoAt day 3 after modeling, the survival rate of the rats in the protective group was higher than that in the control and gavage groups. The differences were statistically significant (P < 0.05); Circled digit threeAfter modeling, the body weight of the rats in the 3 groups were all decreased. The body weight of the rats in each group began to increase from day 7 after modeling. The body weight of the rats in the protective group was all higher than that in the control and gavage groups at day 7, 14, 21, and 28 after the procedure. The differences were statistically significant (P < 0.05); Circled digit fourthere were no significant differences comparing the neurological deficit scores and cerebral infarction volume among the 3 groups. Conclusion: The improved procedures could shorten the time of model production, regain body weight quickly, improve the survival rate effectively after establishment of the cerebral ischemic rat models.

Background Even though the change in wavefront aberrations with correction modality is well documented in the literature,little is known about the underlying mechanism.Complete understanding of the causes responsible for the wavefront change in the combined lens-eye system is important since it provides basic knowledge for further improving the technique to correct refractive error by correcting lenses.Objective The aim of this study was to investigate the influence of refractive correction lens on optical property of the eye by analyzing Zernike aberrations in myopic eyes with contact lens correction.Methods This study was approved by the Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject before entering this study.Zernike aberrations of 52 myopic eyes of 26 subjects with the spherical equivalent-1.75 to-8.50 D were measured using a Hartmann-Shock wavefront sensor.The human eye aberrations were examined at the uncorrected condition,rigid-gas-permeable contact lens (RGP-CL) corrected condition and soft contact lens (Soft-CL) corrected condition.The differences of wavefront aberrations and Zernike coefficients were compared by repeated measurement of single factor variance analysis,and correlation of the aberration changes between uncorrected condition and RGP-CL corrected condition or Sofi-CL corrected condition,between the right eyes and left eyes in different conditions were analyzed by Pearson linear correlation.Results Mean total root-mean-square (tRMS) was (0.71 ± 0.30)μm,(0.54±0.19)μm and (0.74±0.32)μm in the uncorrected condition,RGP-CL corrected condition and Soft-CL corrected condition,with a significant difference (F =8.758,P＜0.001),and tRMS was significant declined under the RGP-CL corrected condition compared with uncorrected condition (t =2.746,P =0.008),and tRMS in RGP-CL corrected condition was significantly lower than that in Soft-CL corrected condition (t =3.428,P =0.001).The high RMS (hRMS) was (0.34±0.12) μm,(0.28 ±0.12) μm,(0.40±0.14) μm in the uncorrected condition,RGP-CL corrected condition and Soft-CL corrected condition,with a significant difference among them (F =10.681,P＜0.001).An insignificant decrease of hRMS was seen in the RGP-CL corrected condition compared with uncorrected condition (t =1.987,P=0.053),but hRMS value was significant higher in the Soft-CL corrected condition than that in the uncorrected condition (t=2.101,P=0.041) and RGP-CL corrected condition (t=4.266,P＜0.001).Compared with uncorrected condition,the axis astigmatism (C5) and spherical aberration (C12) in the RGP-CL corrected condition and spherical aberration (C12) in the Soft-CL corrected condition were significantly reduced (P＜0.05),and the absolute values of trefoil (C6),vertical coma (C7) and tetrafoil (C10) in the RGP-CL corrected condition were lower than those of the uncorrected condition,but vertical coma (C7) absolute value in the Soft-CL corrected condition was increased (P＜0.05).A significantly positive correlation was seen in the spherical aberration (C12) between the RGP-CL corrected condition and uncorrected condition (r =0.763,P＜0.001),and less significant correlation was in the secondary astigmatism (C11) between the Soft-CL corrected condition and uncorrected condition(r=0.469,P＜0.001).Conclusions Different contact lens corrected conditions exert their effects on ocular wavefront structure due to its unique interaction with the eye.RGP-CL wearing has strong modification on wavefront aberrations probably due to its molding effect on corneal surface,which reduces the bilateral symmetry.High order wavefront aberration can be modified by Soft-CL wearing.

Objective To investigate the expression of Foxp3~+ lymphocytes in breast carcinoma tissues and their correlation with other pathological factors,and to investigate the mechanism of action of Treg cells.Methods The expression of Foxp3~+ lymphocytes in the breast cancer tissue and non-cancerous tissue was detected by flow cytometry (FCM) in 30 breast carcinoma patients, and its correlation with other pathological factors was statistically analyzed by multiple linear regression analysis.The expression of TGF-β and IL-10 in the lymphocytes infiltrated in breast cancer tissue and non-cancerous tissue was measured by immunohistochemistry, and their correlation with the expression of Foxp3~+ lymphocytes was statistically analyzed by linear correlation dependability analysis. Results There was significant difference in the expression of Foxp3~+ lymphocytes between the malignant and non-cancerous breast tissues(P<0.05),and it was positively correlated with the clinical stage,blood vessel invasion and the matter of axillary lymph node metastasis(P<0.05). The expression of IL-10 in the tumor infiltrating lymphocytes was positively correlated with the expression of Foxp3~+ lymphocytes(P<0.05).Conclusion The expression level of Foxp3~+ lymphocytes is correlated with invasion and metastasis of breast carcinoma, and the IL-10 secreted by Foxp3~+ lymphocytes may be involved in this effect.Foxp3~+ lymphocytes can be used as an assistant marker for prediction and new therpeutic target of breast cancer.

Animals ; Carcinoma, Hepatocellular ; enzymology ; DNA-Binding Proteins ; Disease Models, Animal ; Liver Neoplasms ; enzymology ; Male ; RNA ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Telomerase ; genetics ; metabolism

OBJECTIVETo explore the effect of arteriosclerosis obliterans (ASO) blood stasis syndrome (BSS) serum on vascular endothelial cell injury and to study the regulation of Taohong Siwu Decoction (TSD) on it.

METHODSUmbilical vein endothelial cell culture system was established. The serum endothelial cell injury model with ASO BSS was prepared. Low, medium, and high concentrations TSD containing serums were respectively added. The endothelial cell proliferation activity was observed by MTT method. Ultrastructures of endothelial cells were observed under transmission electron microscope. Changes of intracellular calcium ion concentration and the cytoskeleton were observed under laser confocal microscope. Contents of ET, NO, and transforming growth factor beta1 (TGF-beta1) in endothelial cell culture supernatant were detected by ELISA.

RESULTSIn ASO BSS serum group endothelial cell proliferation activities decreased, the cell structure was obviously destroyed, calcium ion concentration increased, contents of ET, NO and TGF-beta1 increased significantly (P < 0.01), and ET/NO ratio was imbalanced. After incubating with TSD drug containing serum, endothelial cell proliferation activities and injured cell structures were obviously improved; ET, NO and TGF-beta1 levels decreased (P < 0.05, P < 0.01), ET/NO ratios approximated to the normal level.

CONCLUSIONThe main mechanism of TSD for treating ASO ASS lied in improving injured vascular endothelial cells and endocrine disorder.

Arteriosclerosis Obliterans ; Cell Proliferation ; Drugs, Chinese Herbal ; therapeutic use ; Endothelial Cells ; Humans ; Medicine, Chinese Traditional ; Serum ; Transforming Growth Factor beta1 ; metabolism ; Umbilical Veins

Action Potentials ; Animals ; Atrial Fibrillation ; physiopathology ; Atrial Function, Left ; physiology ; Cardiac Pacing, Artificial ; Pulmonary Veins ; physiology ; Rabbits ; Streptomycin ; pharmacology ; Verapamil ; pharmacology

Aim To investigate the effect of dihydromyricetin on canine carotid artery and the underlying mechanism.Methods The in vitro isometric tension measurement technique was employed to investigate the effect of dihydromyricetin on canine carotid artery rings.Laser scanning confocal microscope technique was used to measure the dynamic change of intracellular calcium concentration in single VSMC.Results Dihydromyricetin(1～300 ?mol?L-1)caused a concentration-dependent contraction of both endothelium-intact and endothelium-denuded rings.This constrictive effect was attenuated in Ca2+-free solution(P

Aim To investigate the effect of hypercholesterolemia on L-type Ca2+(ICa-L) current and intracellular calcium concentration ([Ca2+]i) in single ventricular myocytes of hypercholesterolemic rats.Methods 12 male wistar rats were randomly divided into two dietary groups:a group fed a normal diet(n=6)and a group fed high-cholesterol diet(n=6) for 4 weeks,respectively. The level of serum lipid was examined.Zymolytic method was used to isolate single ventricular myocytes from hypercholesterolemic and normal rats,which were loaded with Ca2+-sensitive fluorescent indicator Fluo-3/AM.[Ca2+]i represented by fluorescent intensity(FI)was measured by laser scanning confocal microscope.Whole cell patch clamp technique was used to record ICa-L.Results There was no significant influence exhibited on TG level.However, the serum total cholesterol(TC)level of hypercholesterolemic rats was much higher than that of model control group; at the test potential of 0 mV, ICa-L decreased from(-8.56?1.29)pA/pF(Control)to(-5.24?0.90) pA/pF(HC)(n=6 in each group,P

Objective To investigate the relationship and clinical significance of blood plasma brain natriuretic peptide (BNP) and Keshan Disease (KD). Methods Seventy KD patients and 30 healthy volunteers in endemic area were investigated with Doppler Echocardiography for the measurement of left ventricular end-diastolic diameter(LVEDD) and left ventricular ejection fraction (LVEF), and the plasma BNP levels were determined with microparticle enzyme immunoassay. Results The BNP levels in plasma in KD patients [(444.61±102.31), (87.21±23.15)ng/L] were significantly higher than that of healthy volunteers [(34.91±15.21)ng/L],the differencesbeing statistical significant (q=39.74,5.82,P<0.01). The BNP levels in chronic KD patients were higher than that of latent KD patients (q=37.62,P<0.01). The plasma BNP levels in KD patients with LVEDD 60 nun [(928.80±134.27)ng/L] were significantly higher than those of patients with LVEDD 55～60 mm [(89.24±52.31)ng/L] and LVEDD<55 nun [(67.14±6.92)ng/L],the differencesbeing statistical significant (q=44.30,48.16, P<0.01), The plasma BNP levels in KD patients with LVEF<35%[(1654.21±421.35) ng/L] were significantly higher than those of patients with 35% ~ 50%[(421.54±112.32)ng/L] and50% [ (81.21±72.85 ng/L)], the differencesbeing statistical significant(q=24.91,72.66, P<0.01), The BNP levels in LVEF 35%～50% were higher than that of 50% (q=11.84,P<0.01). Conclusion The plasma BNP levels were important for the diagnosis, grouping, therapeutic effect and prognostic evaluation of KD.

OBJECTIVETo prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.

METHODSNP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.

RESULTSThree recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.

CONCLUSIONThe recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.

Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Influenza A virus ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology