ObjectiveTo explore the effect of blocking the JAK/STAT signal pathway on the activity of Caspase-3 in the synovial tissue of rheumatoid arthritis rats.MethodsFifty rat models of collageninduced arthritis,which had arthritis index more than 2 were divided into the model group,the low dosage of AG490 group,the medium dosage of AG490 group and high dosage of AG490 group.Inaddition,6 rats were treated as normal control group.Normal saline,AG490 1,5,10 mg·kg-1 ·d-1 were given by intraperitoneal injection.Then the volume claws and pathologic scores of the rat models were recorded and the activity of Caspase-3 in the synovial tissue were compared.The results were analyzed by one-way ANOVA and LSD-t or Tamhane's T2 test.ResultsThe arthritis of the CIA models progressed fast,the volume of the claws and the pathological score of them were significantly higher than those of the control group.At the same time,no Caspase-3 positive express could be detected in the control group,whilethe model group had slightly increased expression.After different dosages of AG490 were applied,the swollen of joints was significantly improved compared with the model group.The histopathological score of the medium AG490 dosage of group and high dosage group(2.7±0.8,1.8+0.9) were remarkably decreased than those of the model group(4.3+1.2),the differences were statistically significant (P＜0.01).In addition,the Caspase-3 expression in the low,medium and high AG490 dosage group ( 1.90±0.15,3.13±0.33,3.56+0.34) was significantly higher than that of the model group(1.48±0.18)(P＜0.05 or P＜0.01 ).ConclusionBlocking JAK/STAT signal pathway can increase the activity of Caspase-3,reduce the excessive proliferation of synovial tissue,and improve arthritis symptoms.

OBJECTIVETo observe the effect of activation of aldehyde dehydrogenase 2 (ALDH2) by ethanol on the expression of c-Jun N-terminal kinase (JNK) in the kidney of diabetic rats.

METHODSEightheen healthy male SD rats were randomly divided into 3 groups (n = 6): normal control group, diabetes group and ethanol + diabetes group. After 8 weeks, 24 h urine samples from rats were collected to detect urinary protein content. The kidney was isolated and the ratio of kidney weight/body weight (index of kidney weight) was detected. The levels of fasting blood glucose, glycosylated hemoglobin serum urea nitrogen and serum creatinine were measured. Morphological changes of renal tissue were observed by optical microscope. The protein expressions of ALDH2 and JNK in renal tissue were detected by Western blot.

RESULTSCompared with the normal control rats, the levels of fasting blood glucose, glycosylated hemoglobin, serum urea nitrogen, serum creatinine and the index of kidney weight were increased markedly in diabetic rats. The expression of ALDH2 protein was decreased, while p-JNK, JNK protein expressions and the ratio of p-JNK/JNK were increased. The morphological observation was shown that the amount of glomerular mesangial matrix were increased, basement membrane were thickened and capillary lumen were narrowed. However,in ethanol + diabetes group, renal function was improved and the damage of renal structure was attenuated. The expression of ALDH2 protein was increased, while p-JNK, JNK and the ratio of p-JNK/JNK were decreased.

CONCLUSIONEnhanced ALDH2 expression can protect kidney in diabetic rats, which may be relevant with inhibitting the activity of JNK pathway.

Aldehyde Dehydrogenase ; metabolism ; physiology ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Diabetes Mellitus, Experimental ; enzymology ; Ethanol ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Kidney ; enzymology ; Male ; Mitochondrial Proteins ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

In order to investigate the effect of N-tosyl-L-phenylalnylchloromethyl ketone (TPCK) and dexamethasone (Dex) on expression of nuclear transcription factor-kappaB (NF-kappaB) in childhood acute lymphoblastic leukemia (ALL) and its significance, so as to provide the experimental basis for corresponding clinical treatment of ALL, in which NF-kappaB is taken as a target. The biotin-streptavidin method was used to detect the expression of NF-kappaB P65 protein and the effects of TPCK and Dex at clinically relevant dosage on activity of NF-kappaB P65 protein in 20 childhood ALL patients. The results indicated that the expression of NF-kappaB P65 protein was strongly diminished and reached to negative level at 2 hours by treatment with 40 micromol/L TPCK, the positive expression of NF-kappaB P65 protein was (2.5 +/- 1.6)%. TPCK had a time-dependent inhibitory effect on ALL cells cultured in vitro. The expression of NF-kappaB P65 protein in ALL cells was strongly inhibited by clinically relevant concentration of dexamethasone 5.0 microg/ml for 24 hours in vitro. The positive expression was (25.0 +/- 3.0)%, there was significant difference, as compared with untreated ALL cells (T=55, P<0.01). It is concluded that TPCK and Dex can inhibit NF-kappaB activity. Inhibition of NF-kappaB activity may be one of the effect mechanism of dexamethasone on ALL cells. Inhibition of NF-kappaB conduction pathway may have a significant value in childhood ALL treatment.
Bone Marrow Cells ; pathology ; Cells, Cultured ; Child ; Child, Preschool ; Dexamethasone ; pharmacology ; Female ; Humans ; Infant ; Leukocytes, Mononuclear ; pathology ; Male ; NF-kappa B ; biosynthesis ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Protein Synthesis Inhibitors ; pharmacology ; Tosylphenylalanyl Chloromethyl Ketone ; pharmacology ; Tumor Cells, Cultured

Triplet surveys were conducted in the city of Xi' an and two villages (one in the vicinity and the other at a distance) in Shaanxi Province in China in October-November (when agricultural activitis were low), 1997, to elucidate nutrient intakes with a focus on possible urban-rural differences. Total food duplicate samples were collected from non-smoking and non-habitually drinking adult healthy women (about 50 subjects per site and 149 in total). The nutrient intakes were estimated from the weight of food items in reference to national food composition tables. On average, the women took 1873 kcal energy, 54 g protein and 37 g lipid per day, with a lipid energy ratio of18.4%. Both excess and insufficient energy intake was observed as a result of food intake analysis and body mass index determination. With regard to minor nutrient intakes, insufficiency was serious in the case of calcium, vitamin A and vitamin B2, but not with iron. Whereas dependency on plant foods for sources of energy and protein was common to the three regions, Xi' an people consumed more animal foods than those in the villages. Intake of fish and shellfish was quite low throughout the three regions. Among the four types of cereals, wheat was consumed most substantially in the three regions and in three meals (except for the village where people essentially did not take lunch in reflection of low agricultural activities), whereas rice was consumed more in Xi' an than in the two villages. Maize consumption was higher in the two villages (especially for breakfast) than in the city.In contrast, foxtail millet (although in small amounts) was taken primarily in Xi'an and only at the time of breakfast.

OBJECTIVETo evaluate the anti-apoptotic effect of aldehyde dehydrogenase 2 (ALDH2) on myocardial ischemia/reperfusion (I/R) injury in diabetic rats.

METHODSNormal male SD rats were divided into normal, diabetes and ethanol (the agonist of ALDH2) + diabetes groups. In the latter two groups, diabetes was induced by an intraperitoneal injection of 55 mg/kg STZ. Four weeks after the modeling, myocardial I/R was mimicked ex vivo, and lactate dehydrogenase (LDH) content in the coronary flow was determined. The activities of caspase-3 and ALDH2 were evaluated, and the expressions of Bcl-2 and Bax mRNA in the left anterior myocardium were detected using RT-PCR.

RESULTSIn diabetic group, LDH release and caspase-3 activity were increased, while ALDH2 activity and Bcl-2/Bax mRNA expression were decreased as compared to those in normal control group. Compared with the diabetic group, ALDH2 agonist ethanol significantly reduced LDH release and caspase-3 activity, increased ALDH2 activity and Bcl-2/Bax mRNA expression.

CONCLUSIONIn diabetic rats, enhanced ALDH2 expression can offer mycardial protection possibly in relation to suppress cell apoptosis.

Aldehyde Dehydrogenase ; metabolism ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Diabetes Mellitus, Experimental ; complications ; enzymology ; Ethanol ; pharmacology ; Male ; Mitochondrial Proteins ; agonists ; metabolism ; Myocardial Ischemia ; enzymology ; etiology ; Myocardial Reperfusion Injury ; enzymology ; pathology ; prevention & control ; Myocardium ; enzymology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Current quality control patterns are limited to industrial application, for most of the natural chemical reference substances are expensive and unavailable. Herein, a method, quantitative analysis of multi-components with single marker (QAMS), was established and validated to simultaneously determine nine ginsenosides (ginsenoside Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rb3, Rd) in P. ginseng and four ginsenosides (ginsenoside Rg1, Rh1, Rb1, Rd) in P. notoginseng. Using ginsenoside Rb1 as the contrast, the relative correction factors (RCF) of the other eight ginsenosides were determined by HPLC-DAD. Within the linear ranges, the values of RCF of ginsenoside Rb1 to ginsenoside Rg1, Re, Rf, Rh1, Rc, Rb2, Rb3 and Rd were 1.400, 1.215, 1.517, 1.801, 0.944, 1.012, 1.143, and 1.135, respectively. The RCF had a good reproducibility in various instruments, chromatographic columns (RSD = 0.30% - 3.9%). According to their RCF, we simultaneously determined nine ginsenosides in P. ginseng only using one marker. In addition, the RCF of ginsenosides were used to simultaneously quantitative analysis of four ginsenosides in P. notoginseng. The results of QAMS method were validated by comparing with that of external standard method, and no obvious significant difference was found.
Chromatography, High Pressure Liquid ; methods ; Ginsenosides ; analysis ; Panax ; chemistry ; classification ; Quality Control ; Reproducibility of Results

OBJECTIVETo observe the role of activation of aldehyde dehydrogenase 2 (ALDH2) on myocardial ischemia/reperfusion (I/ R) injury in diabetic rats.

METHODSDiabetic rat model was simulated by intraperitoneal injection 55 mg/kg streptozotocin (STZ) and divided into diabetes and ethanol + diabetes groups (n = 8). After 8 weeks, myocardial ischemia/reperfusion model was mimicked in vitro. The ventricular dynamical parameters and lactate dehydrogenase (LDH) content in coronary flow were determined. The fasting blood glucose and glycosylated hemoglobin (HbA1c) level were determined by automatic biochemistry analyzer. The ALDH2 mRNA and protein expressions of left anterior myocardium were evaluated by RT-PCR and Western blot.

RESULTSIn contrast to I/R in normal rat, in diabetic rat, left ventricular development pressure (LVDP), maximal rise/fall rate of left ventricular pressure (+/- dp/dtmax) and left ventricular work (RPP) were decreased, left ventricular end diastolic pressure (LVEDP) and LDH release were increased, and ALDH2 mRNA and protein expressions were decreased; compared with I/R in diabetic rat, ALDH2 agonist ethanol significantly promoted the recovery of LVDP, +/- dp/dtmax, RPP, reduced HbA1c level, LVEDP and LDH released, ALDH2 mRNA and protein expressions were increased.

CONCLUSIONIn diabetic rat, the expression of ALDH2 was decreased when heart was subjected to I/R. Enhanced mitochondrial ALDH2 expression in diabetic rat could play cardiac protective role.

Aldehyde Dehydrogenase ; metabolism ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Diabetes Mellitus, Experimental ; complications ; metabolism ; Male ; Mitochondrial Proteins ; metabolism ; Myocardial Reperfusion Injury ; etiology ; metabolism ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Cell-free stem cell therapy has been an issue of concern, but there is no conclusion on how to extract high-quality exosomes. OBJECTIVE: To extract exosomes from human umbilical cord mesenchymal stem cells by using three different methods, and then to screen the optimal method. METHODS: Exosomes were extracted from human umbilical cord mesenchymal stem cells by using the Total Exosome Isolation test kit, Exo Quick test kit and differential ultracentrifugation method, respectively. Then, transmission electron microscopy was used for morphological observations, BCA was utilized to quantify the protein, and western blot assay was applied to detect surface markers CD9, CD81 and CD63. RESULTS AND CONCLUSION: Extraction of exosomes was completed by all the three methods, and round or oval membranous vesicles were observed under the transmission electron microscope. The protein content and purity of exosomes was highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group, and there were significant differences among the three groups (P < 0.05). Under the same protein concentration, surface specific markers, CD81, CD63 and CD9, were expressed highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group. The operating time was significantly lower in the Exobiology Quick kit group compared with the other two groups (P < 0.05). To conclude, despite a longer operating time, the differential ultracentrifugation method is a rational method to extract enough exosomes with relative high purity.

Objective To describe the epidemical characteristics of A (H1N1) influenza identified in the early stage (from May 11 to June 22, 2009) of the epidemic, in mainland China. Methods Epidemical characteristics of 420 confirmed A (H1N1) influenza cases reported from May 11 to June 22, 2009 were analyzed descriptively, including the distribution of age, sex, source of infection, main symptoms and incubation period. Results A total of 77.8% early cases in mainland China were imported from other countries. Three countries including America, Canada and Australia were attributed to 90% of the imported cases. Most of the cases were from 6 months to 73 years old, with 94% of them under 50 years. Most of the symptoms would include fever (81%), cough (40%) and sore throat (35%). The mean incubation period of second-generation cases was 4.3 (4.2±1.5) days. Conclusion Imported cases dominated the total cases in the early stage of the epidemic had similar gender distribution of those from exporting countries. Fever, cough and sore throat were the three main symptoms manifested in influenza cases. 2.5±1.9(1-11)days was found in imported cases between the day of off-board and the onset of symptoms. The incubation period was 4.3±1.7 (1-8) days among the secondary cases.

Objective To explore the relevant factors of prognosis of intracranial aneurysms after microsurgical treatment and risk factors of cerebral vasospasm (CVS). Methods Three hundred and twenty-two patients with intracranial aneurysms, admitted to and received surgical treatment in our hospital from June 2006 to May 2009, were chosen in our study; their clinical data were retrospectively analyzed. Logistic regression analysis was employed to analyze the influences of age, gender, blood pressure level, blood sugar level, operation time, Fisher's grade, Hunt Hess grade and infection on the prognosis of patients with intracranial aneurysms and the risk of CVS. Results Multivariate logistic analysis indicated that age, Hunt Hess grade, Fisher's grade, CVS, infection and lumber puncture times were the independent risk factors influencing the prognosis of patients with intracranial aneurysms (P< 0.05). Hunt Hess grade, Fisher's grade, number of aneurysms, endplate colostomy, lumber puncture times and infection were the independent risk factors of CVS (P<0.05). Conclusion Age, Hunt Hess grade, Fisher' s grade, CVS, lumber puncture times and infection are the independent risk factors affecting the prognosis of patients with intracranial aneurysms, among which, CVS is the most important factor. CVS is mainly affected by Hunt Hess grade, Fisher's grade, number of aneurysms, endplate colostomy, lumber puncture times and infection.