Objective To explore the optimal method of extraction and determination of rhynchophylline and isorhynchophylline in Ramulus Uncariae cum Uncis. Methods A RP-HPLC was performed on a Phenomenex C18 column (4. 6 mm× 150 mm, 5m) at 30℃. The mobile phase consisted of methanol-0. 01 mmol/L triethylamine solution (70: 30, pH 7.0 adjusted by glacial acetic acid) at the flow rate of 0. 5 mL/min. The automatic sample injector was set at 5℃ and the ultraviolet detector was operated at 245 nm. And then, the effect of different extraction condition on the contents of rhynchophylline and isorhyn-ehophylline in Ramulus Uncariae cum Uncis was investigated. ResultsA good linearity of rhynchophylline was in the range of 2. 5μg/mL to 80. 0μg/mL (Y=76. 7170X-0. 0727,r=0. 999 8),the average recoveries were from 99. 84 ％ to 116. 91％, and the RSD (n=6) were less than 8.8 ％. A good linearity of isorhynchophylline was in the range of 2. 0 μg/mL to 80. 0 μg/mL (Y=87. 4729X-0. 3666, r=0.999 7), the average recoveries were from 87.08 ％ to 104. 97 ％, and the RSD (n=6) were less than 7.3 ％. The contents of rhynchophylline and isorhynchophylline in the methanol-extracted solu- tion were approximately ten times as much as those in water-decocted solution. The main factors which had effect on the ex-traction of rhynchophyUine and isorhynchophylline in methanol solution were supersonic time,methanol amount and cold-dousing time, in a decreasing sequence. The best condition selected by orthogonal experiment wasb as follows: extracting the medicinal material with 20-fold volumes of methanol, cold maceration for 24h and supersonic extraction for 1 h. Conclusion The extraction percentage of rhynchophylline and isorhynchophylline in the extraction condition of cold maceration with methanol and supersonic extraction is superior to that in the water-decocting condition. The method is simple, fast and accurate, and it can be used for the quality control of Ramulus Uncariae cum Uncis.

Objective To investigate the biokinetics of 131 I and its absorbed dose in patients with metastatic DTC. Methods Forty?one patients with metastatic DTC who were referred to 131 I therapy were prospectively recruited from September 2009 to September 2011. Scintigraphic images were serially acquired after the ingestion of 131I (3.70-9.25 GBq) to assess the biokinetics. CT was used to measure the masses of the DTC lesions. Calculation of absorbed doses to DTC lesions was based on the Medical Internal Radiation Dosimetry formula of the Society of Nuclear Medicine and Molecular Imaging. Kruskal?Wallis rank sum test was used to compare the differences of absorbed dose among different tumor deposits. Results In total, 59 io?dine?avid metastatic lesions (35 in cervical lymph node, 17 in lung and 7 in bone) in 41 patients were evalu?ated and the mean absorbed dose of DTC lesions was (10.4±6.5) Gy/GBq, with range of 0.1 to 136.2 Gy/GBq. The median absorbed dose was 11.3 Gy/GBq for lymph node metastase, 10.1 Gy/GBq for lung metastases and 9.6 Gy/GBq for bone metastases. There were no statistically significant differences in absorbed doses a?mong the metastases of lymph node, lung and bone ( Z=0.907, P>0.05) . Conclusion A wide inter?lesion variation of absorbed dose to metastatic DTC is observed.

Objective To explore the association between acute ischemic stroke and albumin to creatinine ratio (ACR).Methods A case-control study.During January to December in 2013, 127 acute ischemic stroke patients as case group and 150 controls who were similar with case group in age and gender were recruited in Tianjin Union Medicine Center . According to diabetes , hypertension , cardiovascular diseases and patients without these three diseases , case group was divided into A1, B1, C1 and D1 subgroups, control group was divided into A2, B2, C2 and D2 subgroups in the same way.The first morning urine from participants were collected .Urinary albumin concentration was tested by nephelometry , urinary creatinine was examined by using enzymic method , ACR were calculated(≥30 mg/g as the cutoff value). Then difference of ACR between case and control group was compared , the cutoff value of albuminuria for ischemic stroke patients was analyzed by ROC , and the risk factor of ischemic stroke were analyzed by logistic regression analysis.Results The prevalence of albuminuria in ischemic stroke patients was 38.58%(49).According to analysis of covariance, after adjustment for age, gender, cardiac-disease, diabetes, hypertension, lnACR in case group was significantly higher than control group (3.18 mg/g vs 2.78 mg/g, t=2.13 P=0.03), especially D1 was significantly higher than D2 subgroup (3.01 mg/g vs 2.51 mg/g,t=5.56,P=0.009) .If 19.82 mg/g from ROC analysis was used as the cut-off value, the sensitivity and specificity are 68.5% and 61.7%.The odds ratio ( OR ) of ACR >19.82 mg/g was about 2-fold when compared with ACR<19.82 mg/g adjusted for stroke risk factors , and the OR value is 2.43 in comparison of patients without diabetes , hypertension and cardiovascular diseases .Conclusions Urinary albumin excretion is the independent risk factor of ischemic stroke .The increased urinary albumin has important clinical significance to predict the risk of ischemic stroke for the patients without diabetes , hypertension and cardiovascular diseases.

Objective To establish a rapid monitoring method of the three common bacteria in mice frozen resources, such as embryo, sperm, etc. Methods To extract DNA of the three positive bacteria( Staphylococcus auerus, Klebsiella?pneumoniae and β?hemolyticstreptococcus) , and establish PCR monitoring method of the three positive strains through designing primer and refining PCR condition. Then extract total DNA of the frozen resources, detect the DNA according to the PCR condition of the three positive bacteria, some samples were detect by fluorescence quantitative PCR at the same time. Results ①successfully establish a PCR detection method of the three positive bacteria, the minimum detectable concentration of Staphylococcus auerus, Klebsiella pneumoniae and β?hemolytic streptococcus is 4?19 × 10 -5 ng/μL, 1?98 × 10 -5 ng/μL and 1?07 × 10 -3 ng/μL. ②Proved that the three bacteria doesn ’ t exist in the sample by normal PCR and fluorescence quantitative PCR methods. Conclusions Establising a rapid monitoring method of common pathogens of frozen embryo and sperm in mice.

Objective To investigate the changes of average body weight gain and serum biochemical indexes of C57BL/6J mice (B6 mouse) and their offspings after frozen-thawed embryo transfer of B6 mice.Methods The mice were divided into three groups in this study.In the experimental group I (E-I,30 males and 20 females),2-cell embryos after in-vitro fertilization were collected,and cryopreserved by EFS method,then obtained the offsprings after transplantation of the recovered embryos to oviduct of recipient mice (ICR mouse).In the experimental group II (E-II,26 males and 17 females),when the mice from E-I grew to maturity,the offsprings were obtained from natural mating of mice from E-I.In the control group (20 males and 20 females),the offsprings came from conventional feeding and natural mating.The three groups of mice were raised to 16 weeks old,weighing the body weight at a regular time intervals,and the serum biochemical indexes were obtained from 16-week-old mice.Then the changes of average body weight and serum biochemical indexes of the mice were analyzed.Results The average body weight of E-I mice was significantly higher than that of control group at each week-age (P<0.01).The average body weight of E-II female mice was significantly higher than that of the control group in 12-16-week old mice (P<0.01),but the average body weight of E-II male mice showed no significant differences compared with the control group except for few weeks.The serum biochemical indexes of E-I and E-II mice were changed in all items except for AST,TP and Ca.Conclusions There are some effects on the average body weight gain and serum biochemical indexes of C57BL/6J mice and their offspings after frozen-thawed embryo transfer.

Objective To determine whether the susceptibility to ischemia-reperfusion injury in aged heart is higher than that in adult heart and,if so,to clarify the mechanisms underlying this change.Methods Wister rats(5-or 20-month-old)were randomly divided into 4 groups(6 animals in each group).The rats were subjected to 30 minutes of myocardial ischemia via ligating the left anterior descending coronary artery,followed by 3 hours of reperfusion(Young-MI/R group and Old-MI/R group);A silk suture around the left anterior descending coronary artery was not ligated in young and old rats(Young-sham group and Old-sham group).Myocardial apoptosis was detected by terminal deoxynueleotidyl transferase biotin-d UTP nick end labeling(TUNEL)staining and caspase-3 activity was detected by using a caspase-3 colorimetrie assay.Nitrotyrosine content,a footprint of in vivo ONOO~-formation,and total NO content were determined by ELISA and chemiluminescence method respectively.Results A significantly exacerbated cardiac reperfusion injury was found in Old-MI/R group as evidenced by increased TUNEL positive myocytes[(19.0?2.1)% vs.(14.6?1.7)%],and increased myocardial caspase-3 activity[(436?35)?mool/mg vs.(340?32)?mol/mg] compared with Young-MI/R group(P＜0.05).Aged hearts had a markedly increased basal NOx level compared with young adult hearts.Marked higher myocardial nitrotyrosine content was found in OId-MI/R group[(7.25?0.18)nmol/g]than that in Young-MI/R group[(4.68?0.15)nmol/g] (P＜0.05).Conclusions In aged hearts,high levels of NO might form highly toxie derivant, ONOO~-,and its subsequent nitrified protein.This may attribute to the increased susceptibility of the aged heart to isehemic-reperfusion injury.

Aged ; Aged, 80 and over ; Blood Platelets ; chemistry ; Humans ; Receptor, PAR-1 ; blood ; Receptors, Thrombin ; blood ; Renal Dialysis ; Uremia ; blood ; therapy

OBJECTIVETo investigate the correlation between Chinese medical syndrome type (CMST) and the ID4 gene promoter methylation state in the bone marrow cells of acute myeloid leukemia (AML) patients, and to discuss the correlation between ID4 gene methylation and the occurrence, development of AML.

METHODSThirty-five inpatients or outpatients from the Department of Hematology, the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were recruited as the test group, while 10 healthy volunteers from the health medical center of the Affiliated Hospital, Shandong University of Traditional Chinese Medicine were recruited as the control group. The ID4 gene promoter methylation states were detected using methylation specific polymerase chain reaction (MS-PCR) in the two groups. Inter-group comparison was performed and CMSTs were compared to analyze the correlation between CMSTs and the gene promoter methylation.

RESULTSTwenty-seven AML patients (77.1%) had methylation of ID4 gene, showing statistical difference when compared with the control group (P < 0.01). The ID4 methylation positive rate of ID4 gene promoter methylation was sequenced from low to high as qi and yin deficiency syndrome < inter-accumulation of blood stasis and phlegm syndrome < toxic heat inflaming syndrome, showing statistical difference when compared with the control group (P < 0.05). The peripheral white blood cells and the bone marrow blast cells were higher in ID4 methylation positive patients than in the ID4 methylation negative control patients with statistical difference (P < 0.05).

CONCLUSIONSPatients of inter-accumulation of blood stasis and phlegm syndrome and toxic heat inflaming syndrome were more likely to have ID4 gene methylation. The ID4 methylation positive expression has verified the essence of evil domination in the early AML at the molecular level. It can also reflect the degree of malignancy of AML to some extent.

Adolescent ; Adult ; Aged ; Case-Control Studies ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Promoter Regions, Genetic ; Young Adult

Objective To investigate the changes of calcium /calmodulin - dependent protein kinase Ⅱ (CAMKⅡD) gene expression in 3T3 - L1 preadipocyte diffe-rentiation and TNF - ? regulation of CAMKⅡD gene expression on matured adipocytes. Methods 3T3 - L1 preadipocytes were cultured and differentiated into adipocytes. The levels of CAMKⅡD gene mRNA expression at various times were evaluated by RT-PCR. Matured adipocytes were interfered with 0.1,1.0,10.0 ?g/L TNF - ? and the levels of CAMKⅡD gene mRNA expression were evaluated. Results The levels of CAMKⅡD gene mRNA expression in 3T3 - L1 adipocytes were significantly down- regulated at first day, compared with those at zero day (P0. 05). CAMKⅡD gene mRNA expression decreased significantly at 12 hours after treatment with 10.0 ?g/L TNF-?, treatment of matured adipocytes with TNF - ? did not have any regulation role on it. Conclusions CAMKⅡD gene is involved in the differentiation of adipocytes and related to the etiology of obesity. The changes in the level of CAMKⅡD gene mRNA expression during 3T3 - L1 preadipocyte differentiation may contribute to the differentiation and adipogenesis of adipocytes. It seems that TNF - ? does not have any regulation role on the expression of CAMKⅡD genes.

Objective To investigate the situation of urine albumin measurement of clinical laboratory in Tianjin.Methods Control materials from patient mixed urine samples were made to validate precision in the clinical laboratories in Tianjin.Reference Material ERM-DA470K was prepared as the first external quality assessment ( EQA) sample, and the bias between laboratories was calculated.Then we give some advice about the methods of routine maintenance, calibration, standardized operation, internal quality assessment and so on to the laboratories which were not qualified in the first EQA.Then the second EQA was carried out and CV and bias were culculated.Results 52 clinical laboratories has 12 series of instruments and 17 series of reagents.The precision research showed that most laboratories ( 93.55%) had good precision for urine albumin measurement, while CV of inter-laboratory was great:the range of low level of control sample was 8.91 -43.95 mg/L, 34.46% for CV; and the high level was 36.32 -281 mg/L, 28.51% for CV.Only 36.5% laboratories were qualified in the first EQA. The qualified rate for nephelometry and turbidimetry was higher (55.6%, 42.9%).The qualified rate of trueness verification was 58.6%in the second EQA, and the CV between laboratories was significantly decreased, Inter-laboratory CV of the five samples were:19.83%, 13.57%, 13.41%, 13.08%, 11.37%. The qudified rate for nephelomety and turbidimetry was 71.4% and 56.3%.Conclusions There are a mide variety of measurement systems of urine albumin in Tianjin, and the CV between these systems is great.Clinical laboratory should strengthen the laboratory standardization operation and upgrade calibration testing to improve the testing consistency.