To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid ; methods ; Ginkgolides ; analysis ; Injections ; Limit of Detection ; Salicylates ; analysis ; Tandem Mass Spectrometry ; methods

Objective To investigate the effects of repeated neonatal administration of dizocipline maleate (MK-801), the N-methyl-D-aspartate (NMDA) receptor antagonist, on the expression of NMDA receptor subunits NMDAR 1 (NR1), NMDAR2A (NR2A), NMDAR2B (NR2B) and the protein levels of nerve growth factor (NGF) in neonatal rats. Methods Neonatal Sprague-Dawley (SD) rats were randomly divided into research group and control group, with 15 ani-mals in each group. Rats were administrated subcutaneously with MK-801 or normal saline from postnatal day (PND) 5 to PND14 (0.25 mg/kg, twice a day). The expression levels of NR1, NR2A, NR2B and NGF were examined on PND15, PND42 and PND70 in the prefrontal cortex and hippocampus. Results At PND15 (neonatal period), there were no signifi-cant differences in the expression levels of NR1, NR2A, NR2B and NGF in the prefrontal cortex and hippocampus be- tween the two groups (P>0.05). At PND42 (adolescence), NGF protein levels in the prefrontal cortex was significantly low-er in research group than in control group [(56.19±37.87) vs. (152.54±53.92), P<0.01]. At PND70 (adulthood), the expres-sion of NR1, NR2A in the hippocampus was significantly higher in research group than in control group [NR1:(149.55%± 27.00%) vs. (100.00%±32.08%);NR2A:(171.54%±19.85%) vs. (100.00%±51.04%). P<0.05]. Conclusion Neonatal re-peated treatment of MK-801 increases the expression of NMDA receptor subunits NR1, NR2A in the hippocampus in adulthood while decreases the expression of NGF in the prefrontal cortex in adolescence, suggesting that neonatal block-ade of the NMDA receptor may influence the growth and development of the nervous system.

Objective To analyze the differences between benign and potential malignant small pancreatic cystic lesions.Methods We retrospectively analyzed the clinical and pathological data of asymptomatic patients with pancreatic small cystic lesions and divided them into benign group (including serous cystic neoplasms,lymphoepithelial cyst and pseudocyst) and potential malignant group (including mucinous cystic neoplasms,intraductal papillary mucinous neoplasms and solid pseudopapillary neoplasms).Comparison of clinical data was made between the two groups.Results 46 patients with pathological results were included (22 cases in benign group and 24 cases in potential malignant group).No difference was detected on demographic data and lab results between the two groups.Compared with benign patients,patients in the potential malignant group were more likely to show thicken wall (P =0.000),mural nodule (P =0.000),solid constituents inside the cyst (P =0.001),wall enhancement (P =0.003) and uneven wall on CT scan (P =0.024).The diagnostic sensitivity,specificity and accuracy of the combination of above mentioned CT features for potential malignant diseases were 91.7％,77.3％ and 84.8％,respectively.Conclusions Pancreatic cystic lesions with thicken wall,mural nodule,wall enhancement,solid parts inside the cyst and uneven wall on CT were more likely of potential malignant entities.

Objective To evaluate the risk factors of massive blood loss in resection of giant liver hemangioma.Method The clinical data of 141 patients who underwent giant liver hemangioma resection were retrospectively studied.These data included general physical condition,laboratory tests,radiologic findings,and various surgical parameters.The patients were divided into the massive blood loss group (＞ 1 000 ml,n =27) and the minor blood loss group (≤1 000 ml,n =114).Logistic regression was performed to determine the risk factors of intraoperative massive blood loss.Results The average diameter of the liver hemangioma was significantly greater in the massive blood loss group than that in the minor blood loss group [(21.7 ± 8.5) cm vs.(14.1 ± 5.3) cm,P ＜ 0.05].The incidences of preoperative leukopenia,anemia,thrombocytopenia and prolonged prothrombin time were higher in the massive blood loss group than that in the minor blood loss group (48.1％ vs.16.7％,37.0％ vs.11.4％,25.9％ vs.3.5％,22.2％ vs.3.5％,respectively,all P ＜ 0.05).Hepatic hemangioma with compressed hepatic veins,inferior vena cava and porta hepatis were more frequently found in the massive blood loss group than in the minor blood group (55.6％ vs.14.9％,44.4％ vs.14.0％,55.6％ vs.12.3％,respectively,all P＜0.05).Logistic regression analysis demonstrated a diameter of hemangioma greater than 15 cm was a risk factor of intraoperative massive blood loss during surgical resection.Conclusions Giant hepatic hemangioma may cause disorders in the hematological and coagulation systems.Compression of major hepatic vessels raised technical difficulty and risks in surgery.Hemangioma with a diameter greater than 15 cm was recognized as a high-risk factor of intraoperative massive blood loss.

Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.
Angelica ; chemistry ; Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cytarabine ; administration & dosage ; Humans ; K562 Cells ; Leukemia ; drug therapy ; Liver ; drug effects ; Male ; Mice ; Mice, SCID ; Polysaccharides ; administration & dosage

OBJECTIVETo explore the role of peroxisome proliferator-activated receptor γ (PPARγ) signaling pathway in osteo- blast differentiation of rat bone marrow mesenchymal stem cells (BMSCs) cultured in simulated microgravity.

METHODSRat BMSCs were cultured in simulated microgravity (by rotating clinostat) in the presence of 10 µmol/L pioglitazone, 10 µmol/L GW9662, or both pioglitazone and GW9662, with the cells cultured in normal gravity as the control group. After osteogenic induction for 14 days, the cells were stained with alizarin red for the bone nodules and with oil red-O for the fat cells, and the fat rate was calculated. ALP activity in the cells was determined in each group, and RT-PCR was performed to detect cellular expressions of PPARγ mRNA.

RESULTSPioglitazone significantly inhibited osteoblast differentiation of the BMSCs, whereas GW9662 promoted the cell differentiation by suppressing the activation of PPARγ.

CONCLUSIONWe hypothesize that the activation of PPARγ signaling pathway is one of the main mechanisms for inhibited osteoblast differentiation of rat BMSCs in simulated microgravity, and inhibiting PPARγ pathway activation can effectively prevent and treat microgravity-induced osteoporosis.

Animals ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Osteogenesis ; PPAR gamma ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Weightlessness Simulation

Objective To construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization ( SSH) technique. Methods Suppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3. 1(-)-PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Results The subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones. Conclusion The obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1TP1.

AIMTo prepare capsaicin transfersomes and evaluate them in vitro and in vivo.

METHODSCapsaicin transfersomes were prepared by high shear dispersing machine and evaluated by entrapment efficiency, release rate, in vitro skin permeation and distribution in different tissues in vivo.

RESULTSCapsaicin transfersomes were composed of single unilamellar vesicles with an average diameter of 150.6 nm. Capsaicin entrapment efficiency increased distinctly with increasing of concentration of lecithin and entrapment efficiency is 96.7% while concentration of lecithin to 8%. Cumulative release amount of capsaicin is in direct proportion to the ethanol concentration in the receptor medium. In vitro capsaicin cumulative penetration amount showed higher levels in transfersomes than cream and suspension in rat abdominal skin. Abdominal skin cumulative penetration amount in vitro of capsaicin transfersomes in mouse was significantly higher than that from rat and men. In the same way, abdominal skin epidermal membrane cumulative penetration amount in vitro of capsaicin transfersomes was significantly higher than that from derma and full skin in human abdominal skin. The capsaicin tissue distribution of capsaicin injection by multiple celiac injections in rats is different: bone > plasma > skin > muscle. There is a similar result by multiple thigh topical application of capsaicin transfersomes: bone > skin > plasma > muscle.

CONCLUSIONEntrapment efficiency of capsaicin transfersomes reached the criterion of China Pharmacopoeia (> 80%) and capsaicin skin penetration can be increased by capsaicin transfersomes. It should be noted that the diverse characters and levels of skin may probably affect the permeating capability of capsaicin. Capsaicin tissue distribution in bone and muscle is similar and is different in plasma and skin by multiple injections and topical skin apply.

Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; pharmacokinetics ; Capsaicin ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Humans ; Lecithins ; chemistry ; Male ; Mice ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Sodium Cholate ; chemistry ; Tissue Distribution

BACKGROUNDThe Taq/B, Msp/ and I405V polymorphisms of cholesteryl ester transfer protein (CETP), an important regulatory factor of lipid metabolism, have been attracted much more attention by the researchers. In this study, we investigated the associations between these 3 polymorphisms of CETP gene and variations in plasma lipid and lipoprotein levels in patients with coronary heart disease (CHD).

METHODSGenomic DNA was extracted from leukocytes of 203 CHD patients and 100 control subjects using the salting out method. Genotyping of the CETP gene was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques. Statistical analysis was conducted using the SPSS 10.0 software package.

RESULTSThe distribution of allele and genotype frequencies of the Taq/B, MspI, and I405V polymorphisms was similar in the CHD patient group and the control group. The B1B1 genotype of the Taq/B polymorphism was associated with significantly higher TC (P=0.039) and LDL-C (P=0.044) levels than the B2B2 genotype in CHD patients, and with significantly higher LDL-C (P=0.034) levels than the B2B2 genotype in controls. Homozygotes of the I405V polymorphism exhibited significantly higher HDL-C levels than VV homozygotes among control subjects (P=0.023). In male CHD patients with unambiguously assigned haplotypes, B2-M2-V/B2-M2-I patients demonstrated significantly higher HDL-C concentrations than B1-M2-V/B1-M2-I (P=0.023) and B1-M2-V/B1-M2-V patients (P=0.047).

CONCLUSIONSGenetic variations in the CETP gene may account for a significant proportion of the differences in plasma lipid and lipoprotein concentrations among the general population. The B1B1 genotype of the Taq/B polymorphism is probably a genetic risk factor for CHD in the study population.

Adult ; Aged ; Carrier Proteins ; genetics ; Cholesterol Ester Transfer Proteins ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; genetics ; Female ; Gene Frequency ; Glycoproteins ; genetics ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo evaluate the effect of Valeriana officinalis var latifolia(VOL) on expression of transforming growth factor beta 1 (TGF-beta 1) in hypercholesterolemic rats and study its possible mechanisms.

METHODDietary-induced hypercholesterolemia was induced in male Wistar rats by given 4% cholesterol and 1% cholic acid diet for 16 weeks. Changes of serum lipid, urinary albumin, renal function and Mesangial matrix index were assessed. Moreover, immunohistochemical stain for TGF-beta 1 and type IV collagen were performed.

RESULTVOL could reduce the serum levels of total cholesterol, low density lipoprotein, urinary albumin and serum creatinine. Light microscopy and immunohistochemical stain revealed that in the same time of lowing serum lipid, Mesangial matrix index was significantly reduced, accompanied by decreased expression of TGF-beta 1 and type IV collagen.

CONCLUSIONVOL has the protective effect on lipid-induced nephropathy, and the inhibition of TGF-beta 1 expression might be the mechanism of VOL on renal protection.

Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Hypercholesterolemia ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; Male ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; Valerian ; chemistry