Aim To establish a method for recording tension changes in small artery in vitro.Methods The vessel segments of intestine were dissected from the mesenteric artery of rats and mounted onto Myograph.The internal circumferences of the segments were normalized with Chart5 software.Then the internal diameters of the segments and initial setting of tension were calculated with self-made Excel file.Following these preparation steps, the tension changes induced by several stimulations were recorded by Myograph.Results Normalization procedure provided the data of lumen diameters of small arteries,which was convenient for initial setting of tension and guaranteed the uniformity of the experimental conditions.Myograph could record the tension changes of small arteries induced by several classic stimulations.Conclusion Precise recording method for tension changes in small arteries in vitro is successfully established.

The ancient discussion and modern clinical application regarding Yingxiang (LI 20) and Neiyingxiang (EX-HN 9) in acupuncture verses are explored. Acupuncture verses are the summary of clinical experiences of acupuncture scholars in the past dynasties, of which the records on application of Yingxiang (LI 20) and Neiyingxiang (EX-HN 9) mainly focused on nasal, facial and head-eye diseases, which is characterized with overall summarization, brilliant exposition, simple language, and distinctive characteristics. Nowadays, based on the ancient usage, the clinical application about these two acupoints is continuously developing. If the ancient acupuncture verses could be understood thoroughly, the clinical practice could be better guided and its effects could be improved.
Acupuncture Points ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Rhinitis, Allergic ; therapy

Objective To observe the effect of Xin Guanxin Suhe Drop Pill(XGSDP) on isoprenaline (ISO)-induced acute myocardial ischemia (AMI).Methods Acute myocardial ischemia rats model were established by injection of ISO. Electrocardiogram (ECG) was observed, superoxide dismutase(SOD) activity, and the contents of malondialdehyde(MDA) and nitric oxide (NO) in the myocardial homogenate as well as serum and glutamate oxaloacetate transaminase (GOT) were observed after ISO injection. Results XGSDP could significantly increase NO content and SOD activity, decrease contents of MDA and GOT and reduce the positive rate of ECG( P

Objective:To study the content determination method for the effective components in WudangⅡFlos lonicerae Caulis to lay foundation for the quality evaluation. Methods: An ultrasonic method was used. The effects of extraction solvent, ultrasonic time, ultrasonic power and ratio of solid to liquid on the contents of rutin and mignonette nucleoside were studied, and the extraction conditions were optimized by a 4-factor and 3-level orthogonal experiment. The chromatographic conditions were as follows:a Phenome-nex Luna-C18(250 mm ×4.60 mm, 5 μm) column was adopted for chlorogenic acid, and a Fortis Xi Phenyl column (250 mm × 4. 6 mm, 5 μm) was used for rutin, loganin and luteoloside;the mobile phase was acetonitrile (B)-0. 4% phosphoric acid (C) solu-tion (15 ∶85) for chlorogenic acid and loganin, and acetonitrile (B) -0. 5% glacial acetic acid aqueous solutjion (D) with gradient e-lution for rutin and luteoloside;the column temperature was 30℃, and the detection wavelength was 327,237,354 and 348 nm, re-spectively. Results:The optimum extraction conditions for rutin and luteoloside from WudangⅡFlos lonicerae Caulis were as follows:the extraction solvent was 60% ethanol, the solid-liquid ratio was 1 ∶30, the ultrasonic power and the ultrasonic time were 350 W and 50 min for rutin, and 250W and 60min for luteoloside. The content of chlorogenic acid, loganin, rutin and luteoloside was 10. 27, 6. 33, 0. 401 and 0. 450 mg·g-1 in the samples, respectively. Conclusion:The method is simple and convenient, accurate and re-producible, which can be used to control the quality of WudangⅡFlos lonicerae Caulis and provide reference for the further develop-ment.

Objective To explore the effect of hypoxia inducible factor-1α silencing by siRNA on proliferation, apoptosis and necrosis of human renal proximal epithelial cell ( HK-2 )under hypoxia. Methods CoCl2 was used to mimic hypoxia, and siRNA technique was used to silence HIF-1α. HK-2 cells were divided into five groups, including normal culture group,hypoxia culture group, transfection reagent group, negative control group and HIF-1α siRNA group.MTT assay was used to detect the growth inhibition ratio of cells. TUNEL and caspase-3 quantity was used to detect apoptosis. LDH activity in supernatant was detected to evaluate cell necrosis.Real-time PCR and Western blotting were used to detect mRNA and protein of HIF-1α, HIF-2α,glucose transporter 1(Glut-1) and vascular endothelial growth factor (VEGF). Results (1) The transfection efficiency of siRNA was 95%-100%. Under normoxia, the efficiency of siRNA silencing HIF-1α gene reached to 70%, while under hypoxia, it was 97%. (2) The growth inhibition ratio of cells in HIF-1α siRNA group was significantly higher than that of other groups including hypoxia culture group, transfection reagent group and negative control group (all P＜0.05). No significant difference was found in apoptotic ratio of the other four groups except normal culture group (P＞0.05). The LDH level in HIF-1α siRNA group was significantly higher than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). (3) The expression of HIF1α, Glut-1, VEGF mRNA and protein in HIF-1α siRNA group was significantly lower than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). No significant difference was observed on the level of HIF-2α mRNA and protein in the other four groups except normal culture group (P＞0.05). Conclusion HIF-1α gene silencing can inhibit the mRNA and protein expression of Glut-1 and VEGF, and can aggravate growth inhibition and necrosis of HK-2 cells under hypoxia.

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

Objective To explore the effects of Dimethyloxalyl Glycine(DMOG)on isehemic acute renal failure(iARF)in mice and its relationship with activation of hypoxia inducible factor 1α(HIF-1α).Method Twenty five C57/BL male mice were divided into 5 groups randomly:control group,DMOG group,sham operation group,ischemia/reperfusion(I/R)group and DMOG pretreated group(DMOG+I/R).Ischemia/reperfusion injury was induced in mice by clamping both renal pedicles for 30 minutes.The expression of HIF-1α was determined by Western blot.Renal function was reflected by blood urea nitrogen(BUN)and serum creatinine(Scr).Morphologic changes were evaluated under light microscopy.Apoptosis in the kidney was detected by TUNEL staining.Expression of Vimentin,a marker of tubulointerstitial damage was detected by immunohistochemistry.Results The elvated levels of of BUN((65.8±2.6) vs (13.6±0.7),P＜0.01],and Scr[(229.5±11.2) vs (6.5±0.8),P＜0.01]andwere found morphological injury were induced by the ischemic insult in I/R group.Administration of DMOG dramatically improved renal function[BUN,(26.3±6.5)vs(65.8±2.6);Scr,(27.0±14.1)vs(229.5±11.2),P＜0.01]associated with amelioration of tubulointerstital damage.In the DMOG treated group,the protein level of HIF-1α in the kidney of mile was also up-regulated significantly.Conclusions The protection against iARF in mice by DMOG administration is mediated by activation of HIF-1α.

Objective To analyze the significance of the Calgary Syncope Seizures Scores (CSSS)and the Modified Calgary Syncope Seizure Scores (MCSSS)for differential diagnosis of syncope or epilepsy in children. Methods Totally 201 children[95 male,and 1 06 female,aged 5 -1 8 years,mean age (1 1 .76 ±3.03)years]with syncope or epilepsy who visited the syncope clinic or admitted to the Department of Nerve Specialty Clinic of Pediatrics, the Second Xiangya Hospital of Central South University from October 201 3 to April 201 4 were included in the study. Patients were eligible if they had ≥1 loss of consciousness.The diagnosis was analyzed by the CSSS and the MCSSS and receiver operating characteristic (ROC)curve was used to explore the predictive value of different scores in differential diagnosis of syncope or epilepsy in children.Results There were significant differences in the CSSS be-tween syncope[-4(-6,1 )]and epilepsy[2(-3,5)]in children(Z =-1 1 .63,P <0.01 ).When the score was ≥1 ,the sensitivity and specificity of the differential diagnosis between syncope and epilepsy were 91 .46% and 95.80%, respectively;and Youden index was 0.87.Epilepsy should be considered when the score was ≥1 .There were significant differences in the MCSSS between syncope[-4(-6,1 )]and epilepsy[3(-3,6)]in children(Z =-1 1 .71 ,P <0.01 ).When the score was ≥1 ,the sensitivity and specificity of the differential diagnosis between syncope and epilep-sy were 92.68% and 96.64%,respectively;and Youden index was 0.89.Epilepsy should be considered when the score was ≥1 .Conclusions CSSS and MCSSS might be used as an initial diagnostic method in differential diagnosis be-tween syncope and epilepsy in children,based on the history of the patients.MCSSS in the differential diagnosis between syncope and epilepsy in children was more objective,easier to operate in the clinical work than CSSS.

OBJECTIVETo investigate the cell features of 6 Castleman's disease patients and evaluate their prognosis.

METHODSThe resected tumors were investigated by routine histopathology and immunohistochemistry. Reverse-transcript PCR (RT-PCR) and sequencing of RT-PCR products were used to assess the clonal characters of the main tumor cells.

RESULTSHistologically, all of the 6 tumors were classified as the hyaline vascular type. B-cells dominated the follicular germinal centers, with T-cells dispersing inter-follicularly. The results of RT-PCR each obtained a single band of either 128 bp or 122 bp and sequencing showed that there was highly homogeneity within the same length sequences, accompanied by fewer different nucleotide acids.

CONCLUSIONMonoclonal and/or oligoclonal B cells were identified in Castleman's disease. These B cells were originated from germinal center cells.

Adolescent ; Adult ; Antigens, CD20 ; analysis ; B-Lymphocytes ; metabolism ; pathology ; Castleman Disease ; genetics ; metabolism ; pathology ; Clone Cells ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Humans ; Immunohistochemistry ; Leukocyte Common Antigens ; analysis ; Male ; Prognosis ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the effect of CD40 small interfering RNA(siRNA)on the expressions of pe-ripheral blood interleukin(IL)-21 and IL -35 in rats with experimental autoimmune myocarditis (EAM)and its sig-nificance.Methods Twenty 6 -8 week male Lewis rats were divided into normal group,EAMgroup,CD40 siRNA group and siRNA group by using random number table,with 5 rats in each group.The normal rats were induced with phos-phate buffer saline in double foot pads on day 0 and day 7,while the rest 3 groups were induced with cardiac myosin protein to establish EAMmodels.The rats in CD40 siRNA group and siRNA group were respectively injected with CD40 siRNA and siRNA slow virus expression vector through the tail vein of rats on day 7.The rats were executed on 21 day after echocardiogram examination was made.The histopathologic changes were observed by using light microscope and the myocardial histopathology scores were calculated.Enzyme -linked immunosorbent assay was used to determine the levels of IL -21 and IL -35 in peripheral blood.Results (1)Except the normal group,the total incidence rate of rats of each group was 100%,and there was no rat death.(2)Compared with EAM group,the heart mass/body ratio and myocardial histopathology scores were lower in CD40 siRNA group,and the differences were significant (3.13 ±0.21 vs 3.80 ±0.29,2.22 ±0.43 vs 3.32 ±0.51,F =0.332,0.456,all P <0.05).(3)The echocardiogram showed that there was only 1 rat in EAM group with massive pericardial effusion,and there was no pericardial effusion in CD40 siRNA group.EAMgroup,CD40 siRNA group and siRNA group displayed hypertrophy of the ventricular septum and left ventricular wall,narrow heart cavity and weakening of ventricular wall motion.The left ventricular shortening rate in CD40 siRNA group was significantly higher than that in the EAMgroup[(63.34 ±11.06)% vs (38.56 ±6.98)%,F =16.080,P <0.05].(4)The peripheral blood level of IL -21 in CD40 siRNA group was lower than that in EAM group [(141.19 ±17.46)ng/L vs (157.81 ±17.58)ng/L,F =57.008,P <0.05],while its level of IL -35 was signifi-cantly higher than that in the EAMgroup [(195.96 ±18.26)ng/L vs (174.78 ±13.91 )ng/L,F =31.727,P <0.05].(5)The level of IL -21 in peripheral blood was positively correlated with myocardial histopathology scores in EAM group (r = 0.69,P < 0.05 ),but IL -35 was negatively correlated with myocardial histopathology scores (r =-0.64,P <0.05).Conclusions CD40 siRNA might relieve the myocardial inflammation and reduce the myocar-dial injury of EAMrats.The levels of IL -21 and IL -35 can partly reflect the degree of myocardial injury.The mecha-nism may be related to down -regulating the expression IL -21 and up -regulating the expression of IL -35.