OBJECTIVE: To explore the pathogenic mechanism of liver-yang hyperactivity type of hypertension and to observe the effects of Pinggan Qianyang Formula (PGQYF), a compound of traditional Chinese herbals for calming the liver and suppressing yang, so as to provide experimental evidence for new marker proteins of drug therapy. METHODS: A rat model of liver-yang hyperactivity was prepared with spontaneous hypertensive rats (SHRs) by administration of Aconiti Praeparatae Decoction. Adrenal proteins were separated by 2D gel electrophoresis (2-DE). The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database analysis. RESULTS: The rat model of liver-yang hyperactivity was successfully reproduced, and the PGQYF could decrease the grades of irritability, conjunctival congestion and systolic blood pressure of the rats (P<0.05, P<0.01). After analysis, twelve obviously differentially expressed proteins were found, eight of which were identified. The expression levels of isocitrate dehydrogenase and steroidogenic acute regulatory protein in the untreated group were up-regulated as compared with those in the normal control group, and down-regulated in the treatment group. The expression levels of ferritin light chain, elongation factor Tu, Rho GDP disassociation inhibitor 1, flavin reductase and basic transcription factor 3 in the untreated group were down-regulated as compared with those in the normal control group, and up-regulated in the treatment group. CONCLUSION: Differentially expressed adrenal proteins in SHRs with live-yang hyperactivity are successfully identified. This approach may lay a foundation for the further investigation of pathogenic mechanisms in hypertension with liver-yang hyperactivity and the mechanisms of PGQYF treatment.

Objective To observe the effect of therapy of soothing liver and subsiding yang on hypothalamic protein expression in hypertensive rats with hyperactivity of liver yang,and to explore its therapeutic molecular mechanism for hypertension.Methods Spontaneous hypertensive rats (SHR)were given gastric gavage of Aconiti Praeparatae Decoction (APD)to induce hypertensive rat model with hyperactivity of liver yang,and SD rats served as the normal control.The treatment group received modified Tianma Gouteng Decoction (TGD,0.1g/mL).Two-dimension gel electrophoresis (2-DE)was used to separate the hypothalamic proteins,and the differential expressed proteins were identified by matrix-associated laser desorption-ionization time-of-flight massspectrometry (MALDI-TOF-MS)and database analysis.Results Hypertensive rat model with hyperactivity of liver yang was established successfully.The irritation degree,in-take water volume and systolic pressure were increased in the model group(P

Myelodysplastic syndromes(MDS) is a malignant disease of hematopoietic stem cell / progenitor cell clones. Therapies of MDS including immuno-suppressive drugs, chemotherapy, hemopoietic stem cell transplantation and drugs suppressing malignant clones have improved much in recent years. This review is about intervention therapy of MDS and the therapy effect.

Child ; Humans ; Lung Diseases, Interstitial ; diagnosis

Child ; Child, Preschool ; Enterovirus ; isolation & purification ; Enterovirus A, Human ; isolation & purification ; Female ; Fluorescence ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Male ; Polymerase Chain Reaction ; methods ; RNA, Viral ; analysis

Through analyzing the teaching effect,collected disagreements and suggestions by questionnaire,combining our teaching rules,we proposed efficient innovative methods and suggestions in probation of obstertrics-gynecology.

Objective To perform gene diagnosis of Duchenne muscular dystrophy (DMD) in 3 family members who were negative for DMD gene detected by multiplex PCR and to provide genetic counseling for their family members accordingly .Methods The clinical data and genomic DNA of patients and their family members were collected ,DMD gene mutation were detected by multiplex ligation dependent probe amplification (MLPA) or the 2nd generation of high‐throughput sequencing .Results In the first family ,3 male patients were detected deletion of Exon 7 and 2 female were heterozygous carriers .In the second family ,it was found in the proband that point mutation of c .3127C> T in the Exon 23 of chrX‐32486626 and c .3127C> T heterozygous mutations was confirmed in his mother ,the mother was pregnant with a girl .In the third family ,point mutation of c .2411G>A was detected in the Exon 20 of chrX‐32509581 in the proband and his mother had c .2411G> A heterozygous mutation .Conclusion MLPA or combining with the 2nd generation of high‐throughput sequencing can offer effective gene diagnosis for the patients of DMD and their family members ,and provide the basis for genetic counseling and prenatal diagnosis .

BACKGROUND:Granulocyte colony-stimulating factor (G-CSF) can strongly mobilize bone marrow hematopoietic stem cells (HSCs). It has been proved that G-CSF has the ability to mobilize both HSCs and mesenchymal stem cells (MSCs).OBJECTIVE:To investigate the therapeutic effect of G-CSF in mobilizing autologous bone marrow stem cells entering cerebral infarction zone on ischemic cerebral infarction in rats.DESIGN:A randomized grouping design, animal experiment.SETYING: Center for Stem Cell Biology and Tissue Engineering of Sun Yat-sen University.MATERIALS: This experiment was carried out at the Animal Experimental Department of Sun Yat-sen University (North District) and Center for Stem Cell Biology and Tissue Engineering of Sun Yat-sen University from September 2004 to January 2005. Totally 200 male Wistar rats were chosen and randomly divided into autologous bone marrow stem cells transplantation group and control group, with 100 rats in each group.METHODS:Rats of two groups were made cerebral infarction models by line occlusion. Transplantation group introduced intraperitoneal injection of 60 μg/kg G-CSF one hour after operation. The control group introduced intraperitoneal injection of saline of the same dosage at the same time. ①All rats were weighed before operation and 24 hours, 48 hours, one week after operation to evaluate body mass loss rate. They were also given neurological grading. Grading criteria: Grade 0 is normal. Grade Ⅰ is that the right forelimb bends. Grade Ⅱ is that the right forelimb grasped weakly when the tail is lifted. Grade Ⅲ is that the rat has no directivity in automatic action and circumrotates to right when the tail is lifted. Grade Ⅳ is that the rat circumrotates to right in automatic action. ②15 rats in each group were selected. 24 hours, 48 hours, one week after operation, we opened the skulls, took out the brain and used 2,3,5-Triphenyltetrazoluim Chloride (TTC) staining to measure infarction volume, hematoxylin-eosin(HE) staining to observe the pathological change , and immunohistochemistry to detect the infiltration of CD34+ cells.MAIN OUTCOME MEASURES:Body mass loss rate, neurological grade,infarction volume, pathological change and infiltration of CD34+ cells.RESULTS: Totally 180 of 200 rats were successfully made cerebral infarction model. 48 rats died in seven days after operation. As a result, 132 rat models were alive and 120 rats were randomly selected for data analysis. ①Measurement of body mass and neurological grading: There was no significant difference in body mass loss rate between two groups 24 hours and 48 hours after operation (P ＜ 0.05);one week after operation, body mass loss rate was significantly lower in transplantation group [(10.5±8.2)%]than in control group [(17.8±7.1)%] (P ＜ 0.05). There was no significant difference in neurology grade between two groups. ②Infarction volume:Infarction volume and the percent of infarction volume in the whole brain in control group were all higher than those in the transplantation group,with significant difference [ (251.69±52.77) mm3 vs(145.72±28.05)mm3,(17.00±2.69)% vs (9.90±1.62)% ,P ＜ 0.01]. ③Pathological change: 24 hours after operation, the brain tissue of two groups got classical pathological change of cerebral ischemia infarction. There were some mono-nucleus cells infiltrating in transplantation group while none in control group. 48 hours after operation, most nerve cells disappeared and the glial cells were degenerated. There were many mono-nucleus cells infiltrating in transplantation group while a few in control group. One week after operation, tissues in the infarction zone were liquescent with many monocaryons and lymphocytes infiltrating around them in control group. In transplantation group, part of the infarction zone was plerosised through proliferation of newly born capillaries and glial cells and inflammatory cells were not evident. ④Immunohistochemistry: CD34+ mono-nucleus cells were detected in the ischemic territory in transplantation group 24 hours after operation while none in the brain of other side and control group. There were CD34+ mono-nucleus cells and pyramidate cells with mutations in transplantation group 48 hours after operation while none in the brain of other side and control group.CONCLUSION:The stem cell transplantation in situ therapy, which employs self-marrow stem cells mobilized by G-CSF can relieve the ischemic degree and reduce the infarction volume.

Objective To investigate the clinical therapeutic effect of Xuebijing injection for treatment of patients with diabetic nephropathy(DN)and its mechanism. Methods 60 DN patients were randomly divided into Xuebijing group and control group(each,30 cases). The patients in both groups received western conventional treatment,and the patients in Xuebijing group received additionally Xuebijing injection intra-venous injection once a day for 14 days. The fasting blood glucose(FBG),glycosylated hemoglobin(HbA1c),urinary albumin excretion rate(AER),blood urea nitrogen(BUN),serum creatinine(SCr),hematocrit(HCT),fibrinogen(Fg),whole blood viscosity,total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and interleukin -6(IL-6),tumor necrosis factor-α(TNF-α)and urine β2-microglobulin(β2-MG)levels before and after treatment were detected,and the curative effect was also observed in both groups. Results In the control group blood FBG,BUN,SCr,TC,IL-6 and TNF-αafter treatment were significantly decreased and HDL-C significantly increased compared with those before treatment(all P<0.05). Compared with those before treatment,in Xuebijing group after Xuebijing therapy,blood FBG,β2-MG,AER,BUN, SCr,TC,TG,HCT,blood viscosity,IL-6 and TNF-αwere significantly decreased,and HDL-C was obviously increased,but there were no significant differences in HbA1c,LDL-C and Fg before and after treatment. The above indexes were changed significantly in Xuebijing group compared with those in control group〔FBG(μg/L):6.98±1.14 vs. 9.73±1.62,β2-MG(μg/L):32.1±10.9 vs. 57.2±15.1,AER(μg/min):86.0±28.1 vs. 152.0±51.6,BUN (mmol/L):12.4±8.1 vs. 19.5±8.9,SCr(μmol/L):301.2±151.9 vs. 371.3±168.6,HCT:0.283±0.075 vs. 0.351±0.059,TC(mmol/L):3.4±1.8 vs. 4.1±1.5,TG(mmol/L):3.4±1.5 vs. 3.6±1.7,HDL-C(mmol/L):1.90±0.75 vs. 1.50±0.25, IL-6 (ng/L):8.96±2.07 vs. 12.75±2.47, TNF-α(pmol/L):17.85±4.75 vs. 20.87±4.90,P<0.05 or P<0.01〕. The total efficiency in Xuebijing group was significantly higher than that in control group(83.3%vs. 36.7%,P<0.01). Conclusion Xuebijing injection has significant protective effects on patients with DN,and the mechanism might be associated with increasing tissue perfusion and inhibiting excessive inflammatory cytokines release.

To study the metabolite excretion of theophylline, a rapid and specific method by liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) method for simultaneous determination of theophylline, 1, 3-dimethyluric acid (1,3-DMU), 3-methylxanthine (3-MX) and 1-methyluric acid (1-MU) in human urine was developed using theophylline-d6 and 5-fluorouracil as internal standards. Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the negative mode for mass spectrometric detection. After diluted with methanol and centrifuged, the analytes and ISs were separated on a XDB-Phenyl (150 mm x 4.6 mm, 5 microm) column with a mixture of water-methanol-formic acid (30 : 70 : 0.15) as mobile phase at a flow rate of 0.6 mL x min(-1). The linear calibration curves for theophylline, 1, 3-DMU, 3-MX and 1-MU were obtained in the concentration range of 1.0-250 microg x mL(-1), separately. The method herein described is effective and convenient, and can be used for determination of theophylline and its three metabolites. The results showed that urinary excretion ratio of theophylline, 1,3-DMU, 3-MX and 1-MU is approximately 1 : 3 : 1 : 2 in Chinese subjects, which is similar to the reported excretion pattern in Caucasian.