Adolescent ; Adult ; Brain Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Meningeal Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Meningioma ; diagnosis ; metabolism ; pathology ; surgery ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Recurrence, Local ; Reoperation ; Retrospective Studies ; Tomography, X-Ray Computed ; Vimentin ; metabolism ; Young Adult

OBJECTIVETo evaluate the changes in the concentrations of five components in Fructus Evodiae used in Chinese medicine, including evodiamine and glycyrrhizic acid, during processing of Fructus Evodiae with Radix Glycyrrhizae extract by using high performance liquid chromatography (HPLC) and to provide a scientific basis for different clinical uses of processed and unprocessed Fructus Evodiae.

METHODSThe concentrations of the Fructus Evodiae components in processed or unprocessed Fructus Evodiae were evaluated by HPLC using a YMC J'sphere ODS-H80 column (4.6 mm×250 mm, 5 μm) with acetonitrile-water-tetrahydrofuran-acetic acid (41:59:1:0.2, v/v/v/v) as the mobile phase. The detection wavelength was 225 nm, the column temperature was 35°C, the flow rate was 1.0 mL/min, and the injection volume was 10 μL. The concentrations of the Radix Glycyrrhizae components were determined by HPLC with a Kromasil-C₁₈ column (4.6 mm×250 mm, 4 μm) and a gradient elution of acetonitrile (A) and 0.05% aqueous phosphoric acid (B) as the mobile phase. The detection wavelength was 237 nm, the column temperature was 35 °C, the flow rate was 1.0 mL/min, and the injection volume was 10 μL.

RESULTSThe calibration curves of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin showed good linear relationships (r>0.99). The recoveries of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin were 96.59%, 104.18%, 101.91%, 97.75%, and 97.95%, respectively. The concentrations of the components in processed Fructus Evodiae were obviously different to those in unprocessed Fructus Evodiae.

CONCLUSIONSThe developed method is rapid and accurate. The results provide a reference for processed Fructus Evodiae and the changes that could be expected in its effects compared to unprocessed Fructus Evodiae.

Calibration ; Chromatography, High Pressure Liquid ; Cooking ; Drugs, Chinese Herbal ; analysis ; Evodia ; chemistry ; Glycyrrhiza uralensis ; Plant Extracts ; analysis ; Reference Standards ; Solutions

Carcinoma, Hepatocellular ; metabolism ; Cell Line, Tumor ; DNA, Complementary ; genetics ; GTP-Binding Proteins ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; RNA, Messenger ; genetics ; Transglutaminases ; metabolism

OBJECTIVETo investigate the effects of sympathetic neurotransmitters and adrenergic receptors on liver fibrosis in murine schistosomiasis.

METHODSMice were infestated with schistosoma by means of pasting cercariae on their abdomens. Thirty mice were randomly divided into a control group and a model group. Hematoxylin eosin and Van Gieson staining were used to view the histopathology of their livers. Immunofluorescence histochemistry and laser scanning confocal fluorescence microscopy were used to measure the a1A and beta2 adrenergic receptors in livers of the two groups of mice. High performance liquid chromatography-electrochemical detector (HPLC-ECD) was used to determine the concentration of norepinephrine (NE) and dopamine (DA) in the plasma of the mice.

RESULTSImmunofluorescence histochemistry showed that a1A and beta2 receptors were present in hepatocytes and hepatic sinusoids of the livers of the mice of the two groups, but there were many more in the livers of the schistosoma infected mice (t=-2.888; t=-6.648) (P<0.05). The results of HPLC-ECD showed that the levels of NE and DA in the model group were higher than those of the control group (t=-3.372; t=-4.428) (P<0.05).

CONCLUSIONSympathetic neurotransmitters and adrenergic receptors may participate in liver fibrogenesis in mice infected with schistosoma.

Animals ; Dopamine ; blood ; Liver ; pathology ; Liver Cirrhosis ; metabolism ; parasitology ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; blood ; Norepinephrine ; blood ; Receptors, Adrenergic ; blood ; Schistosomiasis ; metabolism

OBJECTIVE To characterize the molecular weight of Radix Isatidis protein(RIP)and optimize the extraction process of protein from Radix Isatidis.METHODS Response surface methodology(RSM)was applied to optimize the extrac-tion of protein from Radix Isatidis.Extraction time,liquid-to-solid ratio and pH value were set as the investigated factors with respect to the protein yield.In addition,Design Expert software was used for data analysis.The RIP was characterized for composition using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Scanning electron microscopy(SEM)analysis was performed to observe microstructure of the Radix Isatidis powder before and after ultrasound-assisted ex-traction(UAE).RESULTS Based on the RSM analysis,optimum conditions were determined as follows:twice ultrasonic ex-traction in 50 mmol/L Tris-HCl buffer solution,pH at 7.8,liquid-to-solid ratio at 80:1 and extraction for 65 min each time.Under the optimized conditions,the experimental values were 0.705％,which is in close agreement with values predicted by the model.The characterization of the RIP demonstrated that it contained five major groups of protein bands,namely bands of 19.2 kDa,21.5 kDa,24.8 kDa,34～43 kDa and >170 kDa respectively.CONCLUSION RSM can be applied for the optimi-zation of extraction process of RIP,which is effective,stable and feasible.

OBJECTIVETo evaluate the mRNA and protein expressions of peroxiredoxin II (PrxII) in hepatocellular carcinoma (HCC) and their significance.

METHODSHCC was induced by aflatoxin B1 (AFB1) in 6 tree shrews (Tupaia belangeri chinensis). The expression levels of PrxII mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot on HCC tissues and on their surrounding liver tissues (para-HCC). Biopsied liver tissues were taken before the HCC induction (pre-HCC) from the same animals and from a group of blank controlled animals that served as controls. Liver biopsy specimens from 18 cases of human HCC and from 17 healthy human volunteers were studied using the same methods.

RESULTSThe mRNA and protein expressions of PrxII in tree shrew HCC tissues were significantly higher than those in para-HCC and pre-HCC tissues, and also higher than those in the liver tissues from the control animals (all P < 0.05). The expression levels of PrxII mRNA and protein in human HCC tissues were also significantly higher than those in their para-HCC tissues and in the human normal liver tissues (P < 0.05).

CONCLUSIONPrxII might play an important role in hepatocarcinogenesis and might be used as a molecular target for HCC prevention and treatment.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Middle Aged ; Peroxiredoxins ; genetics ; Tupaiidae

This study was aimed to investigate the molecular genetic basis and serological phenotype of Rh weak D type 15 individuals. Samples were identified by serological tests and genotyped by sequence specific primer-PCR (SSP-PCR), and were sequenced to detect the changes of all ten RHD exons. The number of gene RHD was detected through SSP-PCR. The results showed that in tested individuals of weak D type confirmed by the IAT, 18 cases (56% in weak D) were weak D type 15. Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E+e; Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E-e+; others (78%) were c-c+E+e+. The results by serological tests were consistent with the results genotyped by PCR-SSP method. In all 18 samples, the sequencing result revealed a gene mutation 845G > A at the exon 6 of the RHD and the point mutation changed amino acid G282D of the RhD polypeptide. The zygosity test demonstrated that 2 out of 18 weak D type 15 individuals were RHD(+)/RHD(+) homozygous (two DCe/DcE), 16 cases were RHD(+)/RHD(-) heterozygous (two DCe/dce and fourteen DcE/dce). It is concluded that Weak D type 15 is predominant in weak D individuals of Chinese Han Nationality, and most of them are heterozygous with various RH haplotypes.
Asian Continental Ancestry Group ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Erythrocytes ; immunology ; Exons ; genetics ; Genotype ; Haplotypes ; Humans ; Molecular Sequence Data ; Phenotype ; Point Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics ; immunology ; Sequence Analysis, DNA

Carcinoma, Hepatocellular ; metabolism ; Glypicans ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

To study the genetic polymorphism of HPA 1-16 platelet antigen alleles among unrelated volunteer donors and establish a typed platelet donor panel in Handan, typing was perfomed by polymerase chain reaction using sequence-specific primers (SSP-PCR); 148 random unrelated blood donors in Handan were genotyped for each of the HPA 1-16 antigen. The gene frequencies were analyzed and the genetype frequencies were determined by direct counting, and these data were compared with HPA distribution among various population by the chi-square test. The results indicated that HPA-1a, 2a, 4a-14a, 16a genes were found among the 16 HPAs in every sample tested. Monomorphic HPA-4a, 7a-14a, 16a were found in the samples. For HPA-1, 2, 5 and 6, a/a homozygosity was predominant with frequencies of 0.9595, 0.8108, 0.9865, 0.9797, respectively, and none of HPA b/b was found in the samples. HPA-1b, 2b, 5b, 6b were rarely found among subjects. HPA-15 had the greatest heterozygosity with a gene frequency of 0.2230, 0.5270, 0.2500 for HPA15a/15a, HPA15a/15b, HPA15b/15b, respectively. HPA-3 showed the second greatest heterozygosity with a gene frequency of 0.3851, 0.5135, 0.1014 for HPA3a/3a, HPA3a/3b, HPA3b/3b, respectively. HPA genotype frequencies showed a good fit to Hardy-Weinberg equilibrium. HPA1-5 gene frequencies for Chinese people in Handan were consistent with those of Chinese people in Shijiazhuang (P > 0.05). Among the HPA1-13, -15, the frequencies of HPA-1, -2, -6 for Chinese people in Handan differed appreciably from those for Chinese people in Taiwan (P < 0.05), others were similar to those of Chinese people in Taiwan. Among the HPA 1 - 8, a similarity was noted between Chinese people in Handan and Koreans (P > 0.05), except for HPA-3. Frequencies of HPA-1, -2, -5 significantly were differed from those in African Americans, as compared with HPA 1-5 (P < 0.05). Comparison of gene frequencies from HPA-1 and -5 showed significant differences between Chinese people in Handan and people in UK (P < 0.05). It is concluded that HPA-2, -3, -5, -15 of people in Western region of China have polymorphism, incompatible frequency of HPA antigen distribution is higher, which inevitably results in the increase of immunologic exposure, therefore attention must be paid to the importance of HPA-2, -3, -5, -15 in clinical disorders. This study for the first time completely analyses HPA1-16 gene frequencies in China, and provides data for establishing a typed platelet donor panel in Handan, China.
Antigens, Human Platelet ; classification ; genetics ; Blood Donors ; statistics & numerical data ; China ; Gene Frequency ; Genotype ; Humans ; Platelet Transfusion ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae