Objective To establish an effective UHPLC-ESI-MS/MS method for the simultaneous determination of 19 active components (ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside) of different types in Xintiantai I (XI), and provide a comprehensive and efficient quality control method for traditional Chinese medicine (TCM). Methods The analysis was performed on an Agilent 1290 system with a Agilent Zorbax Eclipse Plus C18 column (150 mm × 4.6 mm, 3.5 μm) at a flow rate of 0.4 mL/min using acetonitrile and 0.1% formic acid aqueous solution as mobile phase. Mass spectrometric detection was performed on multiple reaction monitoring (MRM) in positive and negative ionization mode. The contents of 19 active components in XI were determined by monitoring the specific ions of each component. Results The 19 active components were accurately determined in 15 min and had the good linearity (r2 > 0.999) within the linear ranges. The average recovery rates of ginsenoside Rg1, Rb1, Rd, berberine, epiberberine, jatrorrhizine, palmatine, columbamine, coptisine, evodiamine, rutaecarpine, dehydroevodiamine, limonin, hyperin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, echinacoside, and verbascoside were 94.80%, 96.78%, 95.59%, 96.88%, 97.74%, 100.08%, 96.27%, 100.25%, 98.32%, 97.16%, 95.60%, 95.28%, 96.81%, 95.22%, 96.85%, 95.31%, 93.86%, 94.79%, and 95.20%, respectively; The contents of three batches XI of the 19 components were in the ranges of 2.28-2.49, 0.82-0.90, 1.22-1.32, 14.44-15.50, 3.71-3.99, 3.26-3.49, 3.09-3.33, 4.39-4.72, 4.56-4.92, 0.52-0.57, 0.30-0.33, 4.46-4.76, 3.02-3.24, 2.59-2.76, 6.03-6.38, 1.47-1.58, 1.90-2.08, 3.40-3.88, and 1.53-1.74 mg/g, respectively. Conclusion The developed UHPLC-ESI-MS/MS-MRM method is fast, sensitive, and reproducible for TCM quality control. It can be used for the quality control of XI, which also provides reference for TCM quality research.

AIM To establish an HPLC-ELSD method for the simultaneous content determination of gastrodin,parishin A,parishin B,ginsenosides Rg1,Re,Rf,Rb1 and Rd in Tiantai No.1 Tablets (Ginseng Radix et Rhizoma,Gastrodiae Rhizoma,Cistanches Herba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 45 ℃C thermostatic Alltima C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrie-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner.RESULTS Eight constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.94％-97.93％ with the RSDs of 1.06％-2.48％.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Tiantai No.1 Tablets.

OBJECTIVETo explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.

METHODSSix clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.

RESULTS1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).

CONCLUSIONThere is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Osteoporosis ; genetics ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley

Previous methods used for nuclear transplantation were further investigated to develop a method that was both easy to carryout and did not require any special apparatus, such as Piezoimpact or Spindle-View. Following the puncture of zona pellucida with two holes by injection pipette that contained donor nuclei or cells, the injection pipette was pulled back to the perivitelline space while the negative pressure was increased in the holding pipette until the polar body and karyoplasm were wiped off completely. Then a reconstructed embryo was completed by the direct injection of the donor nucleus or cell without pulling out the injection pipette. 200 oocytes were manipulated using this method and it cost about 40 seconds with nucleus injection and about 30 seconds with cell injection to complete a reconstructed embryo. The success rates were 62.6% and 86. 0%, respectively, and enucleation rate was about 73.3% validated by Hoechst 33342. Using this method, the nucleus was completely eliminated and another was injected using the microscope and micromanipulator. Moreover, the efficiency of nuclear transplantation and survival rate of reconstructed embryos were greatly improved. Furthermore, it is very easy to manipulate and popularize in practice.
Animals ; Cell Culture Techniques ; methods ; Cell Nucleus ; metabolism ; Cells, Cultured ; Cloning, Organism ; methods ; Embryo, Mammalian ; cytology ; metabolism ; Embryonic Development ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mice, Inbred Strains ; Nuclear Transfer Techniques ; Oocytes ; cytology ; metabolism ; Zona Pellucida ; metabolism

OBJECTIVETo investigate the influence of persistent rapid atrial pacing on the levels of connexin 43 (Cx43) and type III collagen in pulmonary vein and atrium in a canine model.

METHODSSixteen mongrel dogs were divided into rapid atrial pacing (RAP) group (n = 8) and normal control group (n = 8) randomly. In the RAP group, atrial pacing was performed with a rate of 400 bpm for 10 weeks to establish atrial fibrillation model. The tissues of left superior pulmonary vein (LSPV), left atrial free wall (LAFW) and right atrial appendage (RAA) were collected from each dogs. The levels of Cx43 and type III collagen were measured in each tissue.

RESULTSTen weeks later, persistent atrial fibrillation was induced in all dogs in RAP group. The level of Cx43 in RAP group was higher than that in normal control group (LSPV: 3370.91 +/- 275.11 vs 1405.82 +/- 90.38, P < 0.05; LAFW: 2448.68 +/- 272.10 vs 1467.12 +/- 147.93, P < 0.05, RAA: 2331.96 +/- 199.61 vs 1288.27 +/- 216.22, P < 0.05). The level of Cx43 in LSPV was higher than that in LAFW and RAA in RAP group, whereas the difference between LAFW and RAA was not significant in RAP group. The quantities of type III collagen in RAP group were higher than those in normal control group (LSPV: 3301.97 +/- 309.70 vs 1404.56 +/- 178.02, P < 0.05; LAFW: 2477.86 +/- 190.43 vs 1479.20 +/- 187.17, P < 0.05; RAA: 2045.92 +/- 139.43 vs 1417.07 +/- 139.43, P < 0.05). The quantities of type III collagen in LSPV was higher than those in LAFW and RAA in RAP group.

CONCLUSIONSPersistent rapid atrial pacing could increase the levels of Cx43 and type III collagen in pulmonary vein and atrium in a canine model of atrial fibrillation. The levels of Cx43 and type III collagen in pulmonary vein were higher than those in atrium. This findings indicated that pulmonary vein may be a crucial regions in maintaining atrial fibrillation.

Animals ; Atrial Fibrillation ; metabolism ; physiopathology ; Cardiac Pacing, Artificial ; methods ; Collagen Type III ; blood ; Connexin 43 ; blood ; Disease Models, Animal ; Dogs ; Female ; Male ; Pulmonary Veins ; metabolism ; physiopathology

OBJECTIVETo study the efficacy and safety of multimodal prevention of postoperative venous thromboembolism for hip fractures.

METHODSFrom March 2009 to July 2011, preoperatively, patients were assigned to two groups on the basis of an assessment of their risk factors. One hundred and twelve patients were considered to be low risk, involving 47 males and 65 females,with an average age of (72.40 +/- 13.29) years ranging from 42 to 88,and were managed with aspirin (100 mg once daily for 14 days) as well as intermittent gasing compression devices. Twenty-six patients were considered to be high risk, involving 12 males and 14 females with an average age of (78.50 +/- 12.76) years ranging from 65 to 84,and were managed with low-molecular-weight heparin (0.4 ml,subcutaneous injection once daily for 14 days) and intermittent gasing compression. All patients were underwent Doppler ultrasonography within 24 hours before hospital discharge. All patients were followed-up for 3 months postoperatively. The incidence of deep venous thrombosis of lower limb, pulmonary embolism, gastrointestinal hemorrhage were recorded.

RESULTSOverall, there were no fatal pulmonary embolism, 1 case of symptomatic pulmonary emboli in low risk group, and none were detected in the high-risk group. Deep venous thrombosis was detected in association with 6 (6.25%) of the 112 procedures in the low-risk group and 2 (7.69%) of the 26 operations in the high-risk group. Paitents were selected in opened reduction and internal fixation, the quantity of bleeding, decrease of hemoglobin, hematoma rate, and gastrointestinal hemorrhage rate of low risk group were (538.10 +/- 390.20) ml, (30 +/- 19) g/L, 0, and 1 (1.03%) respectively; those of the high-risk group were (585.95 +/- 403.96) mL, (32 +/- 20) g/L,1 (4.76%), (4.76%), there were no significant different between the two groups, all P > 0.05.

CONCLUSIONThere were no statistic significances between the aspirin as well as intermittent gasing compression devices and the low-molecular-weight heparin and intermittent gasing compression in preventing venous thromboembolism (VTE) in postoperative postoperative venous thromboembolism for hip fractures. However, there are potential advantages to reduce complications of bleeding and cardiovascular disease. Multimodal prevention of postoperative venous thromboembolism can protect postoperative patients with hip fractures.

Adult ; Aged ; Aged, 80 and over ; Female ; Hip Fractures ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Risk ; Venous Thromboembolism ; prevention & control

OBJECTIVESTo study the contribution of the genes and environment to variation of serum levels of lipids and lipoprotein.

METHODSOne hundred and forty-three healthy monozygotic (MZ) twin pairs and 93 dizygotic (DZ) ones aged 5 to 19 [with a mean of (11.2 +/- 3.4) years]. Microsatellite polymorphism (STR) was used to diagnose zygosity of twins, and intraclass correlation coefficient method and Falconer formula were performed to investigate heritability of serum lipids and lipoproteins unadjusted or adjusted for age and sex. Logarithmic transformation was used for data with skewed distribution. Influence of relevant physical and biochemical indicators on serum lipids and other components was analyzed with partial coefficients of correlation adjusted for age and sex.

RESULTSIn the twin samples, difference in serum level of triglycerides (TG) between MZ and DZ was not statistically significant with intraclass variation and intraclass correlation. There was significant difference in serum levels of total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and lipoprotein [Lp(a)] between MZ and DZ, with their heritability estimates of 0.56, 0.55, 0.49 and 0.58 unadjusted, respectively, and 0.63, 0.63, 0.55 and 0.64 adjusted for age and sex, respectively. Serum levels of TC, HDL-C, LDL-C and Lp(a) correlated reversely with age. Serum levels of TC, HDL-C and LDL-C in girls were slightly higher than those in boys. Most indicators for serum levels of lipids and lipoprotein, except for serum level of Lp(a) correlated with body mass index (BMI), body fat ratio, Pelidisi index, and other indexes such as blood pressure, blood sugar, serum level of calcium, adjusted for age and sex.

CONCLUSIONSSerum levels of TC, HDL-C and Lp(a) were influenced more greatly by genetic factors, and serum level of TG was mainly influenced by environmental ones. Levels of blood lipids in children were influenced by age and sex, and correlated with indicators that reflect their body fat and nutritional status.

Adolescent ; Adult ; Age Factors ; Child ; Cholesterol, HDL ; blood ; genetics ; Cholesterol, LDL ; blood ; genetics ; Female ; Humans ; Lipids ; blood ; genetics ; Lipoproteins ; blood ; genetics ; Male ; Sex Factors ; Triglycerides ; blood ; genetics ; Twin Studies as Topic ; Twins, Dizygotic ; Twins, Monozygotic

Objective To observe the urodynamic change after spinal cord injury at different levels and the relationship with neurogenic dysfunction of bladder and urethra. Methods Eighty female rats were divided into a control group (20 rats) , a suprasacral spinal cord injury group (30 rats) and a sacral spinal cord injury (30 rats). The urodynamic exam was performed with all the rats before and 20 days after the spinal cord injury model was established by surgical operation. Results The maximum bladder volume and compliance in the su- prasacral injury group were significantly less than the sacral spinal cord injury group and the control, the maxi- mum volume and compliance in sacral spinal cord injury group were significantly less than the control. The DLPP in suprasacral injury group was significantly higher than that in the sacral spinal cord injury group and the con- trol, the DLPP in sacral spinal cord injury group was significantly less than that in the control group. Conclu- sion Urodynamic study is very useful for the early diagnosis and individualized treatment of the neurogenic bladder after spinal cord injury.

Objective To study the effect of Radix astragali injection (RA) on the activities of cytochrome P450 2D6(CYP2D6),2C19,3A4,1A2 and 2C9 by ultrahigh performance-LC and tandem mass spectrome try(UHPLC-MS/MS).Methods The activities of CYP2D6,2C19,3A4,1A2 and 2C9 were evaluated based on pharmacokinetics of metopmlol,omeprazole,midazolam,phenacetin and tolbutamide (five probe drugs) respectively.SD rats were assigned to two groups:control group and experimental group and the rats in the two groups were given 0.9％ NaCl and RA 10 mL · kg-1 by intraperitoneal injection for consecutive 7 days respectively.On the 7th day,after intraperitoneal injection for 30 min,a mix dose of five probe drugs was intragastric administration to all rats.Blood samples were collected at different times for UHPLC-MS/MS analysis.Pharmacokinetic parameters of five probe drugs were assessed between the two groups respectively.Results For metoprolol,omeprazole,phenacetin and tolbutamide,the AUG0-t in RA group decreased by 37.8％,55.5％,40.9％ and 38.9％ respectively,the Cmax in experimental group decreased by 25.4％,33.6％,14.6％ and 40.2％ respectively.While the AUC0-t and Cmax of midazolam in experimental group increased significandy by 60.1％ and 195％ respectively.RA could increase the total amount of midazolam absorbed into blood,while decreasing the total amounts of metoprolol,omeprazole,phenacetin and tolbutamide absorbed compared with control group respectively.The method of validation conformed to the requirements of the relevant rat pharmacokinetic guidehnes.Conclusion This method was successfully employed to assess the effect of RA on the activities of CYP2D6,2C19,3A4,1A2 and 2C9 in vivo.RA could inhibit the activity of CYP3A4,while inducing the activities of CYP2D6,2C19,1A2 and 2C9.

Objective To explore the risk factors of type 2 diabetes mellitus(T2DM)in mountainous countryside residents in order to provide relevant intervention measures.Methods 80 new cases of T2DM were selected from pingyuan community in Liandu District of Lishui City as case group,and 1 60 normal people were selected as control group at the same time and places,in which the 1∶2 matched design of case -control study method was used.Conditional Logistic regression analysis was applied in this study.Results The multiple conditional logistic regression analysis suggested that primary family history,history of hypertension,abdominal obesity were the risk factors of T2DM(OR=2.08,1.55 and 2.78 respectively);while education level,exercise and fish intake were the protective factors(OR=0.74,0.59 and 0.32 respectively).Conclusion T2DM is the result of both genetic factors and environmental factors.Moreover,health education,balanced diet and proper physical exercise should be encouraged among residents to prevent T2DM in mountainous countryside.