The dissolution of Panacis Quinquefolii Radix ultrafine granular powder and common powder, traditional pieces in water and simulated gastric juice in vitro was compared, and the effect of particles size of Panacis Quinquefolii Radix on the dissolution was studied. HPLC method was used for determination of five ginsenosides including Rg1, Re, Rb1, Rc and Rd from ultrafine granular powder and common powder, traditional pieces of Panacis Quinquefolii Radix at different points in time, furthermore, the dissolution curves of Panacis Quinquefolii Radix ultrafine granular powder and common powder, traditional pieces were obtained. The dissolution characteristics of the three Panacis Quinquefolii Radix forms were also compared in this study. According to the results, the dissolution rates of ginsenosides from ultrafine granular powder exceeded 90% of the total content with 5 min, significantly higher than that of the other two forms in water in vitro. At the same time, the dissolved amount of the ultrafine granular powder was fourteen percent higher than that of the traditional pieces and eight percent higher than that of the common powder. Under the condition of simulated gastric juice in vitro, the dissolution rates of ginsenosides from ultrafine granular powder were little lower than that of the other two, but the maximum dissolved amount of the former was fourteen percent higher than that of the common powder and five percent higher than that of the extracts. Therefore the conclusion is that micronization of Panacis Quinquefolii Radix contributed to dissolution of effective components.
Chromatography, High Pressure Liquid ; Ginsenosides ; chemistry ; Panax ; chemistry ; Plant Roots ; chemistry ; Powders ; Solubility

This study aims to analyze and compare the effect of cell wall-broken decoction pieces, conventional decoction pieces and conventional powder of Rhodiolae Crenulatae Radix et Rhizoma on the intestinal flora of normal mice. The conventional bacterial culture and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) were adopted for the mice after the oral administration for 14 days. According to the bacterial culture results, the 1/8 dose cell wall-broken decoction pieces group showed fewer Enterococcus and Escherichia coli bacillus but more Lactobacillus and Bifidobacterium than the conventional decoction pieces group and the traditional powder group (P <0.05). Meanwhile, on the basis of the PCR-DGGE results, the 1/8 dose cell wall-broken decoction pieces group revealed the highest Shannon-Wiener index (H) and species richness (S) among the seven groups, with extremely significant differences compared with the normal group (P <0.01), significant differences compared with the conventional decoction pieces group and the conventional powder group (P <0.05) and a high intra-group similarity. In conclusion, the long-term intake of 1/8 dose Rhodiolae Crenulatae Radix et Rhizoma cell wall-broken decoction pieces showed a certain effect in regulating intestinal tract by promoting the growth of Lactobacillus and Bifidobacterium. Furthermore, the intestinal flora community will become more stable.
Animals ; Bifidobacterium ; drug effects ; genetics ; growth & development ; Cell Wall ; Denaturing Gradient Gel Electrophoresis ; Intestines ; microbiology ; Lactobacillus ; drug effects ; genetics ; growth & development ; Mice ; Mice, Inbred C57BL ; Polymerase Chain Reaction ; Rhizome ; Rhodiola

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; analysis ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Powders ; Quality Control

OBJECTIVETo study the infected information, clinical symptom and molecular epidemiological characteristics of HuCV infection among children under 5 years old in Nanjing.

METHODSIn Nanjing Children's Hospital of Nanjing Medical University from July 2010 to June 2011, we collected 428 stool specimens from children with diarrhea and 428 asymptomatic controls. Human Calicivirus were tested by using RT-PCR. Then we sequenced the nucleic acid of PCR amplifications and identified the genotype and gene group of prevalent strains.

RESULTS63 (14.72%) out of 428 stool samples were detected as HuCV. 58 were norovirus and 5 were sapovirus, while GII-4 2006b was the predominant strain of NoV. In the 428 control samples, 19 samples were positive for calicivirus, there were 8 NoV and 13 SaV (Including 3 co-infection cases).

CONCLUSIONHuman caliciviruses with different genotypes circulated among children in Nanjing,and GII. 2006b is the dominant genotype.

Caliciviridae ; classification ; genetics ; isolation & purification ; Caliciviridae Infections ; epidemiology ; virology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Molecular Epidemiology ; Phylogeny ; Seasons

OBJECTIVETo establish a nude mouse model of malignant ascites with human ovarian carcinoma cell line OVCAR3 which highly expresses VEGF and evaluate the therapeutic of Avastin combined with cisplan.

METHODSForty-eight nude mice with malignant ascites resulting from intraperitoneal transplantation of human ovarian carcinoma cell line OVCAR3 were treated with intraperitoneal injection of Avastin, cisplan, their combination, and PBS, respectively, to observe the effect on ascites development, VEGF content in the ascites, peritoneal permeability, development of new vessels and number of tumor cells in the ascites.

RESULTSAvastin obviously inhibited ascites accumulation and peritoneal capillary permeability, reduced VEGF protein level and microvascular density in the tumor tissues and the number of red cells and tumor cells in the malignant ascites, and prolonged the survival of the mice. The combination of Avastin and cisplan further enhanced the therapeutic efficacy of Avastin.

CONCLUSIONThe bio-chemotherapeutic strategy with Avastin combined with cisplan can be a promising method for treatment of malignant ascites.

Animals ; Antibodies, Monoclonal ; administration & dosage ; Antibodies, Monoclonal, Humanized ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Ascites ; etiology ; metabolism ; prevention & control ; Bevacizumab ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; Drug Synergism ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; pathology ; prevention & control ; Ovarian Neoplasms ; complications ; genetics ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Xenograft Model Antitumor Assays

Objective:To investigate the short-term efficacy and safety of albumin-bound paclitaxel combined with nedaplatin followed by concurrent radiotherapy in treatment of stage Ⅲ massive cervical cancer.Methods:The clinical data of 84 patients with massive cervical cancer admitted to Harbin Medical University Cancer Hospital from April 2019 to April 2020 were retrospectively analyzed. According to the different treatment regimens, patients were divided into the observation group and the control group, each with 42 cases. The observation group received albumin-bound paclitaxel combined with nedaplatin followed by concurrent radiotherapy, and the control group received solvent-based paclitaxel combined with nedaplatin followed by concurrent radiotherapy. The short-term efficacy and adverse reactions of the two groups were compared.Results:The partial remission (PR) rate of the observation group and the control group at 1 month of treatment was 92.9% (39/42) and 35.7% (15/42), respectively, and the difference was statistically significant ( χ2 = 29.867, P < 0.01). The complete remission (CR) rate of the observation group and the control group at 1 month after treatment was 59.5% (25/42) and 38.1% (16/42), respectively, and the difference was statistically significant ( χ2 = 3.859, P = 0.049). The incidence of diarrhea of the observation group was lower than that of the control group [33.33% (14/42) vs. 54.8% (23/42)], and the difference was statistically significant ( χ2 = 3.913, P = 0.048). There were no statistical differences in the incidence of hematological adverse reactions and abnormal liver and kidney functions between the two groups (all P > 0.05). Conclusion:The albumin-bound paclitaxel combined with nedaplatin followed by concurrent radiotherapy have a good short-term efficacy in treatment of stage Ⅲ massive cervical cancer, and the adverse reactions are tolerable.

BACKGROUNDT-lymphoma and metastasis gene 1 (Tiam1) produces a guanine nucleotide exchange factor (GNEF) that regulates guanosine triphosphatase, which transforms guanosine diphosphate to guanosine triphosphate. Recently published data indicate that Tiam1 was associated with gastric cancer. The aim of this study was to investigate biological effects and potential mechanisms of Tiam1 in gastric carcinoma.

METHODSWe analyzed the expression of Tiam1 in 114 pair-matched gastric neoplastic and adjacent non-neoplastic tissues by quantitative real-time PCR. We investigated Tiam1 expression and its prognostic value for gastric cancer. Furthermore, the functions of Tiam1 over-expression were analyzed with stable-expression Tiam1 plasmid in human gastric cancer cell lines.

RESULTSTiam1 expression was significantly associated with cell differentiation and lymphatic metastasis; expression of Tiam1 mRNA was up-regulated in gastric cancer compared to pair-matched adjacent non-tumor tissues. Analyses of surgical tissue samples and 5-year survival of gastric cancer patients showed that those with strong Tiam1 expression had significantly shorter overall survival time than those with negative Tiam1 expression. Ectopic expression of Tiam1 promoted cell growth, migration and invasion of gastric cancer cells in vitro.

CONCLUSIONSIn gastric cancer cells, Tiam1 affects multiple properties associated with acquisition of the metastatic phenotype, and may be a marker of gastric cancer progression and metastasis in a subset of cancer.

Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Cell Proliferation ; Guanine Nucleotide Exchange Factors ; genetics ; metabolism ; Humans ; Neoplasm Metastasis ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; T-Lymphoma Invasion and Metastasis-inducing Protein 1

OBJECTIVETo compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model.

METHODSFive New Zealand white rabbits (2 to 3 kg, either gender) were isolated nucleus pulposus and annulus fibrosus under sterilized condition, then cultured in nutrient solution with 15% FBS and DMEM/F12 (1:1) by enzyme digestion combined with tissue block method. When 90% cells fused, subcultring were performed. Cell morphology were observed by inverted phase contrast microscope, cell viability were detected by trypan blue staining, histological were observed by a toluidine blue and HE staining, cell proliferation were tested by MTT method, then the cell morphology, viability, proliferation between nucleus pulposus and annulus fibrosus were compared.

RESULTSThere were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later, and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day.

CONCLUSIONThe cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus,local biomechnical changes, furether caused structure change and function loss of annulus fibrosus.

Animals ; Cell Proliferation ; Cell Survival ; Disease Models, Animal ; Female ; Humans ; Intervertebral Disc ; cytology ; Intervertebral Disc Degeneration ; physiopathology ; Male ; Rabbits

OBJECTIVETo explore the effect of pro-angiogenic factors and their receptors on angiogenesis in hepatocellular carcinoma.

METHODSExpression of VEGF/KDR and Angiopoietins/Tie2 was detected by RT-PCR and Western blot in 15 cases with hepatocellular carcinoma, 15 tumor adjacent tissues (<1 cm, >5 cm), 8 cirrhotic liver, and 4 normal liver. Immunohistochemistry (IHC) was used to detect CD34 expression, and the relationship between neovascular density and angiogenesis was analyzed.

RESULTSThe expression levels of VEGF and Ang2 were significantly higher in hepacellular carcinoma group than those in the other groups (P < 0.01), and so did the expression of CD34. The expressions of KDR and Ang1/Tie2 showed no significant difference in all groups, but they indeed increased to various levels in tumor and tumor adjacent tissues as compared with those in cirrhosis and normal liver.

CONCLUSIONVEGF/KDR and Angiopoietins/Tie2 may be the crucial signal pathways in the development of hepatocellular carcinoma.

Adult ; Aged ; Angiopoietin-2 ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Middle Aged ; Neovascularization, Pathologic ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, TIE-2 ; biosynthesis ; genetics ; Signal Transduction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; genetics

OBJECTIVETo screen the gene expression profiles of IFN-alpha antiviral proteins based on a low-density cDNA Macroarray, and to explore the relationship between the expression of antiviral protein and the HBV replication.

METHODSThe HepG2 and HepG2.2.15 cells were treated with various concentrations of IFN-alpha (0 IU/ml, 100 IU/ml, 1000 IU/ml) of IFN-alpha for 6 h, and then the low-density cDNA Macroarray was used for analysing the expression profiles of antiviral genes and screening differential expressions of antiviral proteins. Meanwhile, the HepG2 cells were transiently transfected with HBV core protein-expressed plasmid pHBc-EGFP, and the expressions of antiviral proteins were analysed by RT-PCR assay. Moreover, the HepG2.2.15 cells were also transfected with the antiviral protein-expressed plasmid pcDNA3.1-Flag-MxA. ELISA was used for analysing the secreted HBV antigens, while dot blot and Southern blot were applied for analysing the extracellular HBV DNA and intracellular replicative intermediate HBV DNA in HepG2.2.15 cells. All data were presented as mean+/-SD and analyzed using the t-test and one-way analysis of variance (ANOVA) in the experiments.

RESULTSThe Macroarray results suggested that the expression of IFN-alpha antiviral genes like 6-16, IFITM1, IFITM2, IFITM3 and RING4 in HepG2.2.15 cells were partially inhibited. More importantly, it was found, in this research, the expression of antiviral protein MxA in HepG2.2.15 cells was completely suppressed. RT-PCR analysis indicated that the expression of MxA was also significantly decreased in HepG2 cells transfected with pHBc-EGFP plasmid. Although HepG2.2.15 cells transfected with pcDNA3.1-Flag-MxA plasmid could not inhibit extracellular HBV DNA and intracellular replicative intermediate HBV DNA, the MxA exerted some antiviral activities as it effectively suppressed the secretion of HBsAg and HBeAg in HepG2.2.15 cells.

CONCLUSIONSHBV and its antigen components probably influence the expression of antiviral proteins. IFN- resistance may be related to the down-regulation of antiviral proteins expression.

Antiviral Agents ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Viral ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Interferon-alpha ; pharmacology ; Plasmids