Objective To investigate the effects of ketamine on the expression of NMDA receptor-1(NRⅠ)in a rat model of focal cerebral ischemia and the possible mechanism of the neuroprotection.Methods Forty healthy male SD rats weighing 250-290g were randomly divided into 2 group(n=20 each):groupⅠketamine and groupⅡpentobarbital.The aminals were anesthetized with intraperitoneal ketamine 60 mg?kg~(-1) in groupⅠor pentobarbital 40 mg?kg~(-1) in groupⅡ.Focal cerebral ischemia was produced by permanent middle cerebral artery occludion(MCAO).The animals were killed at 24 h and 72 h of MCAO and their brains removed for determination of infarct size,the number of living neurons in the penumbra and the expression of NRⅠprotein(immuno- histochemistry).Results The infarct size was significantly smaller;the number of living neurons in penumbra significantly larger and NRⅠexpression significantly down-regulated in ketamine group than in pentobarbital group.Conclusion Ketamine can protect the brain against ischemia through downregulation of NMDA receptor-1.

Flammulin, an anti-tumor protein, was purified from the aqueous extract of basidiomes of Flammulina Velutipes. Purified flammulin emulsified with Freund's adjuvant was injected subcutaneously into New Zealand white rabbits. After several immune enhancements, these animals were bled and sera were separated. Antiserum against flammulin in Western blots were applied to determine if flammulin be present in the liquid state culture or fruiting body. The result showed that anti-flammulin serum could recognize the aqueous extract of fruiting body in SDS-PAGE gels under the reducing conditions, no flammulin was detected in mycelia of Flammulina Velutipes.

Purpose To assess the efficacy and prognostic factors of alpha-fetoprotein (AFP) negative hepatocellular carcinoma (HCC) by transcatheter arterial chemoembolization (TACE). Materials and Methods 67 AFP negative HCC patients and 67 AFP positive HCC patients underwent TACE, the survival of two groups was compared, and the prognostic factors were also analyzed. Results All 134 HCC patients were followed up for 24 months, the one-year, two-year and five-year survival rate of AFP negative and AFP positive HCC patients were 86.6%, 58.2%, 31.3%; and 81.6%, 37.8%, 13.4%, respectively, with median survival time of 34.0 months and 19.0 months, respectively. The survival rate were statistically different between AFP negative and AFP positive HCC patients (P<0.001). In AFP positive group, the median survival time of patients with AFP>400 ng/ml and AFP ≤400 ng/ml were 18.0 months and 31.0 months, respectively with statistical difference (P<0.05). Univariate analysis showed that Child-Pugh classification, histological grade, tumor size, tumor number, portal vein tumor thrombus, BCLC staging and AFP level were independent risk factors influencing the prognosis of HCC patients. Conclusion AFP negative and AFP positive (AFP ≤400 ng/ml) HCC patients achieves good efficacy and prognosis by TACE, Child-Pugh classification, portal vein tumor thrombus, BCLC staging and AFP level were risk factors influencing the prognosis of HCC patients.

To optimize indices of molecular identification for authentication of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, four indices, including sequence similarity, specific positions, genetic distance and phylogenetic tree, were compared based on trnL-trnF sequences. Total DNA was extracted from Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, and trL-trnF sequences were amplified and sequenced. Sequence similarity was calculated by BLAST analysis. Specific positions were compared by DNAman software. Genetic distance and phylogenetic tree were analyzed by Mega software. The results showed that the inter-specific and intra-specific similarity of P. ginseng and P. quinquefolius respectively was 100% and 99. 6%. There were four specific positions at G153A, T463A, C732G and T818C. The inter-specific genetic distance (0) of trL-trnF sequences was lower than intra-specific genetic distance (0. 004). P. ginseng can be distinguished from P. quinquefolius based on the phylogenetic tree. It is concluded that Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix can be authenticated by identification indices of sequence similarity, specific positions, genetic distance and phylogenetic tree. Index of specific positions based on trnL-trnF sequences is the most efficient index to authenticate Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix.
Chloroplasts ; genetics ; DNA Barcoding, Taxonomic ; methods ; Panax ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Rhizome ; classification ; genetics

Objective To investigate the effects of saponins from Allium Macrostemon Bunge Bulbs (SMBB) on platelet aggregation and platelet-neutrophil-interactions. Methods The effects of SMBB on platelet aggregation in SD rats were observed in vivo and in vitro. The adhesion of platelets to neutrophils was measured by rosette test. The effect of SMBB on activated platelet calcium levels was detected. Results Platelet aggregation induced by platelet activating factor (PAF), adenosine diphosphate (ADP) and arachidonic acid (AA) was significantly inhibited by SMBB in vitro concentration-dependent. Platelet aggregation induced by PAF, AA and ADP was significantly inhibited by 5 mg/kg of SMBB. SMBB could reduce the intracellular calcium concentration in the wash platelets. SMBB significantly reduced the adhesion between neutrophils and thrombin-activated platelets and inhibited neutrophil supernatant-induced platelet aggregation, with IC50of 2.7 μmol/L and 9.6 μmol/L, respectively. Conclusion SMBB can inhibit platelet aggregation in vitro and ex vivo, and inhibit the interactions between platelets and neutrophils.

The fruit of Lycium ruthenicum is a common folk medicine in China. Now it is popular for its antioxidative effect and other medical functions. The adulterants of the herb confuse consumers. In order to identify a new adulterant of L. ruthenicum, a research was performed based on NCBI Nucleotide Database ITS Sequence, combined analysis of the origin and morphology of the adulterant to traceable varieties. Total genomic DNA was isolated from the materials, and nuclear DNA ITS sequences were amplified and sequenced; DNA fragments were collated and matched by using ContingExpress. Similarity identification of BLAST analysis was performed. Besides, the distribution of plant origin and morphology were considered to further identification and verification. Families and genera were identified by molecular identification method. The adulterant was identified as plant belonging to Berberis. Origin analysis narrowed the range of sample identification. Seven different kinds of plants in Berberis were potential sources of the sample. Adulterants variety was traced by morphological analysis. The united molecular identification-origin-morphology research proves to be a preceding way to medical herbs traceability with time-saving and economic advantages and the results showed the new adulterant of L. ruthenicum was B. kaschgarica. The main differences between B. kaschgarica and L. ruthenicum are as follows: in terms of the traits, the surface of B. kaschgarica is smooth and crispy, and that of L. ruthenicum is shrinkage, solid and hard. In microscopic characteristics, epicarp cells of B. aschgarica thickening like a string of beads, stone cells as the rectangle, and the stone cell walls of L. ruthenicum is wavy, obvious grain layer. In molecular sequences, the length of ITS sequence of B. kaschgarica is 606 bp, L. ruthenicum is 654 bp, the similarity of the two sequences is 53.32%.
Berberis ; classification ; cytology ; genetics ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Drug Contamination ; Drugs, Chinese Herbal ; isolation & purification ; standards ; Lycium ; classification ; cytology ; genetics ; Medicine, Chinese Traditional ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity

This study was aimed to explore the effect of BCL11A gene on transcription of γ-globin gene in K562 cells. B-cell lymphoma/leukemia 11A (BCL11A) gene was silenced by small interfering RNA (siRNA) expression vectors in K562 cells (human erythroblastic leukemia cell line). Gamma-globin mRNA level in K562 cells was determined by RT-PCR. Association between the BCL11A gene and γ-globin gene transcription was explored by comparison of mRNA levels. The results indicated that the silence rate of the BCL11A gene in K562 cells by 4 siRNA expression vectors was 49.7%, 55.4%, 78.2%, and 84.1%, respectively. The siRNA expression vector with 84.1% silence rate was transfected into K562 cells, transcription level of γ-globin mRNA in K562 cells transfected with siRNA expression vector increased 2.4 times as compared with control K562 cells. It is concluded that level of γ-globin mRNA increases when the BCL11A gene is silenced. It indicates that the BCL11A gene may be a negative regulator for γ-globin gene expression.
Carrier Proteins ; genetics ; Gene Expression Regulation, Leukemic ; Genes, Regulator ; Genetic Vectors ; Humans ; K562 Cells ; Nuclear Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transcription, Genetic ; Transfection ; gamma-Globins ; genetics

OBJECTIVETo explore the effects of prenatal exposure to low level lead on learning and memory of rat's offspring.

METHODSThe pregnant rats were randomizedly divided into 4 groups and provided with doubly evaporated water in the control group and 125, 250 and 500 mg/L lead acetate solution via drinking water in three exposed groups respectively during the pregnancy. The learning and memory ability of 21-day old and 60-day old offsprings were tested by the Morris water maze and the shuttling and avoiding dark box respectively.

RESULTSThe blood and hippocampus lead concentrations of 1-day old and 21-day old offspring in the 3 lead-exposed groups were significantly increased compared with the control group (P < 0.05), while at the period of 60-day old there was no significant difference between them. The time to find the platform in Morris water maze for 21-day old and 60-day old offsprings of the 3 lead in the exposed groups was significantly increased compared with the control group during the 4 days training (P < 0.05). The times of initiative avoiding in the shuttling and avoiding dark box for 21-day old and 60-day old offsprings in the 3 lead-exposed groups were significantly decreased, while the latency of the initiative avoiding and latency of the passive avoiding were significantly increased compared with the control group (P < 0.05).

CONCLUSIONPrenatal exposure to low level lead impaired the space learning and memory ability of offsprings of the rats, and this kind of influence will continue till the offspring's maturity.

Animals ; Dose-Response Relationship, Drug ; Female ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Maze Learning ; drug effects ; Memory ; drug effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo summarize the experience of hepatectomy for patients with centrally located primary liver cancer.

METHODSThe clinical data of patients with centrally and non-centrally located primary liver cancer were retrospectively reviewed. The biochemical indicators, operation duration, hepatic inflow occlusion time, hospital stay, operative blood loss, amount of blood transfusion, complication, and effectiveness of three occlusion methods (semi-hepatic inflow occlusion, Pringle's manoeuvre, and modified Pringle's manoeuvre) were analyzed.

RESULTSTumor diameter, Child-Pugh score, indocyanine green retention rate, aspartate aminotransferase, alanine aminotransferase, glutamyltransferase, total bilirubin, direct bilirubin, albumin, prealbumin, cholinesterase, hepatic inflow occlusion time, blood transfusion, postoperative complications, and operative blood loss were not significantly different between patients with centrally and non-centrally located primary liver cancer. Patients with centrally located liver cancer had significantly longer operation duration and hospital stay than patients with non-centrally located liver cancer (P < 0.05). The modified Pringle's manoeuvre of hepatic inflow occlusion had the same effectiveness of the Pringle's manoeuvre and could be performed in a simpler way.

CONCLUSIONSHepatectomy is safe and feasible for patients with centrally located primary liver cancer. Appropriate preoperative evaluation and preparation, sufficient knowledge of liver anatomy, and proper selection of hepatic inflow occlusion method are key factors to guarantee the success of the resection.

Adult ; Case-Control Studies ; Female ; Hepatectomy ; methods ; Humans ; Liver Function Tests ; Liver Neoplasms ; blood supply ; complications ; physiopathology ; surgery ; Male ; Middle Aged ; Postoperative Complications ; etiology ; therapy ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the splice variants of the calpain 3 gene existing in human skeletal muscle tissue and white blood cells, and to explore the feasibility of gene diagnosis using CAPN3 mRNA extracted from peripheral leukocytes.

METHODSTotal RNA was extracted from peripheral blood and skeletal muscle tissue in healthy individuals. CAPN3 cDNAs were determined by reverse transcriptase polymerase chain reaction and DNA sequencing. CAPN3 cDNAs from peripheral leukocytes were compared with sequences obtained from skeletal muscle tissue.

RESULTSRT-PCR and DNA sequencing showed that the CAPN3 cDNAs comprised 24 exons in human skeletal muscle tissue, while the number of exons was 23 in white blood cells. Exon 15 was spliced out in human white blood cells.

CONCLUSIONSplice variants exist in human skeletal muscle tissue and white blood cells. Gene diagnosis may omit the mutations of exon 15 using mRNA extracted from peripheral leukocytes. These findings suggest that mutation analysis of the CAPN3 cDNA should use skeletal muscle tissue as materials instead of peripheral blood.

Calpain ; genetics ; DNA Mutational Analysis ; DNA, Complementary ; genetics ; Exons ; genetics ; Humans ; Leukocytes ; metabolism ; Muscle Proteins ; genetics ; Muscle, Skeletal ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction