OBJECTIVE To investigate the protective effect of Codonopsis Pilosula Polysaccharide (CPPS) on improving of the memory consolidation disorder induced by Cycloheximide and its possible mechanisms in mice. METHODS The mice was divided into five groups, as normal control group, cycloheximid model group, piracetam positive control group, CPPS 300 mg · kg- 1 group, and CPPS 150 mg·kg-1 group. The mice respectively were given saline, piracetam, and CPPS for 15 d. The memory consolidation disorder model in mice was established by ip. Cyclohexylamine, and orally administered CPPS(300 mg·kg-1 or 150 mg·kg-1) every day. Then experimental groups were subjected Morris Water Maze test. Western blotting analysis were used to analysis the expression of CaMKⅡ/CREB signaling pathways. RESULTS Morris water maze experiment showed that cyclohexylamine can cause memory consolidation disorder(P<0.01), and giving piracetam and CPPS (300 mg · kg- 1) can improve spatial memory impairment in mice(P<0.05, P<0.01). Western blotting experiment results show that compared with normal control group, CaMKⅡ and CREB contents of brain in model group mice had significant decreased(P<0.001); Compared with model group, CaMK Ⅱ and CREB contents of brain tissue in piracetam and CPPS groups increased significantly(P<0.05,P<0.01,P<0.001). CONCLUSION Cyclo?heximide can induce the memory consolidation disorder, and its effect in mice related to CaMK/CREB signaling pathways. CPPS can improved this memory disorder by influence CaMKⅡ/CREB signaling pathways.

Mongolian folk medicine resource is the origin of Mongolian medicine development, even more important of which is the specialized Mongolian folk medicine resources with regional and high medicine quality, it processes distinctive national characteristics with irreplaceable important position in traditional Mongolian medicine. Nevertheless, due to the serious destroy of ecological environment and sharp increase of demands, etc. A lot of specialized Mongolian folk medicine resources were endangered, and there still existed some problems in the protection and exploitation and utilization. This paper intends to provide comprehensive insight into the species protection and exploitation and utilization states of specialized Mongolian folk medicine resources. The application and protection status and the existing problems were reviewed, and the development strategies of Mongolian folk medicine resource were analyzed.
Conservation of Natural Resources ; methods ; Environment ; Medicine, Mongolian Traditional ; Mongolia ; Plants, Medicinal ; classification ; growth & development

OBJECTIVETo use distinctive water-medicine cup therapy for treatment of cervical spondylopathy of cervical type as a sample to provide a more effective cupping method for clinic.

METHODSOne hundred and forty cases were randomly divided into a medicine cup group (n = 47), a water cup group (n = 47) and a empty cup group (n = 46). The medicine cup group were treated by cupping therapy with a self-made medicine cup with 45 degrees C Chinese herb solution; the water cup group were treated with a cup with 45 degrees C water, and the empty cup group with a cup with nothing. Clinical symptoms and signs were observed for comparison of therapeutic effects.

RESULTSClinically cured was 39 cases, and markedly effective was 8 cases in the medicine cup group; 20 cases were clinically cured, 22 cases were markedly effective and 5 cases were effective in the water cup group; 12 cases were clinically cured, 19 cases were markedly effective and 15 cases were effective in the empty cup group. There were significant differences in the ratio of cases of different therapeutic effects and the difference of pain score before and after treatment between the medicine cup group and the water cup group (P < 0.05), between the water cup group and the empty cup group (P < 0.05), and between the medicine cup group and the empty cup group (P < 0.01).

CONCLUSIONThe therapeutic effect of the distinctive medicine cup is better than the water cup group, and the water cup group is better than the empty cup group.

Adolescent ; Adult ; Aged ; Cervical Vertebrae ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Spinal Osteophytosis ; therapy

OBJECTIVESTo investigate a new targeting mechanical arm for CT-based navigated percutaneous fixation of pelvic fractures, and to evaluate the safety and efficiency of the procedures.

METHODSUsing CT-based 3D navigation software combined with targeting mechanical arm, percutaneous insertion of pelvic models (3 dry human cadaver pelvic skeletons and 5 plastic Sybone pelvic models) were performed, 8 pelvic models allowed percutaneous cannulated screw insertion of both S-I joint (2 S-I screws placement for each side, total 32 screws in this experiment) and both superior ramus (1 ramus medullary screw placement for each side, total 16 screws in this experiment). Percutaneous insertion of pelvic models (4 dry human cadaver pelvic skeletons and 4 plastic Sybone pelvic models, 1 S-I screws and 1 ramus medullary scre placement for each side, 32 screws in this experiment) were performed using fluoro-navigation system (Stryker, USA). Time necessary for every screw insertion were recorded. Accuracy of screw placement was assessed using C-arm imaging and direct eyes inspecting. The time and accuracy of the two methods were compared.

RESULTSThe time required for the CT-based 3D navigation procedure (3.6 ± 1.2) min was significantly less than using the targeting mechanical arm compared to drilling freehand with navigation (9.1 ± 0.8) min (t = 2.50, P < 0.01). There was no significant difference in accuracy between the two methods.

CONCLUSIONCT-based 3D navigation software combined with targeting mechanical arm should be potential to apply percutaneous sacroiliac screwing for pelvic fractures with more accurate and more reliable.

Bone Screws ; Cadaver ; Fracture Fixation, Internal ; methods ; Humans ; Models, Anatomic ; Pelvic Bones ; surgery ; Software ; Surgery, Computer-Assisted ; methods

ObjectiveTo investigate the potential pharmacological mechanism of Xinmaikang tablets in the treatment of atherosclerosis cardiovascular disease by using network pharmacology and cell experimental validation. MethodThe components of Xinmaikang tablets were searched by BATMAN-TCM database and the active ingredients and potential targets were screened. The atherosclerosis related disease targets were searched in GeneCards and online mendelian inheritance in man（OMIM） disease databases. The therapeutic targets were obtained by mapping the intersection of the tablets and disease targets. Therapeutic targets were uploaded to STRING database to construct protein-protein interaction（PPI） network. Cytoscape software was used to create a "drug-active component-therapeutic target" network map， and a network topology algorithm was used to screen key action targets. David software was used for gene ontology（GO） and Kyoto encyclopedia of genes and genomes（KEGG） function enrichment analysis. The key targets of drug therapy were validated by in vitro cell assay. ResultA total of 19 active ingredients， 132 potential targets and 4703 atherosclerotic disease-related target genes of Xinmaikang tablets were retrieved and screened， and 84 intersection targets were obtained. 3 key therapeutic targets of Xinmaikang tablets in the treatment of atherosclerotic diseases were screened， including Calmodulin 1（CALM1）， voltage-dependent L-type calcium channel subunit alpha-1C（CACNA1C） and Phosphatidylinositol 4，5-bisphosphate 3-kinase catalytic subunit alpha isoform（PIK3CA）. A total of 313 biological processes， 89 molecular functions and 53 cell components were obtained by GO enrichment. A total of 40 pathways were obtained from KEGG functional enrichment， including purine metabolism， renin secretion， CGMP/PKG signaling pathway and so on. In vitro cell experiment results verified that Xinmaikang tablets can up-regulate the expression of CALM1 and CACNA1C， down-regulate the expression of PIK3CA， so as to inhibit the activity of inflammatory response， and play a therapeutic role in atherosclerotic diseases. ConclusionXinmaikang tablets may treat atherosclerosis cardiovascular disease through betulin， methyleugenol and other compounds， through purine metabolism， renin secretion， cGMP/PKG signaling pathway and other pathways， which acts on CALM1， CACNA1C， PIK3CA and other targets.

OBJECTIVEThrough comparison of HER2/neu oncogene detected by chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) in breast cancer, to explore the effect of CISH on detecting gene amplification of HER2.

METHODSSelected formalin-fixed paraffin-embedded breast samples whose pathological types were infiltrating ductal carcinomas (255 retrospective samples, 271 prospective samples), and these samples were detected by IHC and CISH.

RESULTS(1) In the retrospective study, CISH identified gene amplification in 91.6% of IHC score 3+ tumors (120/131) and in 56.5% of IHC score 2+ tumors (39/69), thus the concordant ratio between IHC and CISH was 81.2% (207/255). The two results showed significant correlation (P<0.01). (2) In the prospective study, the ratio of HER2 protein over expression detected by IHC was 31.7%, the ratio of HER2 gene amplification detected by CISH was 27.3%. CISH identified gene amplification in 91.4% of IHC score 3+ tumors (53/58) and in 46.4% of IHC score 2+ tumors (13/28), Concordant ratio between IHC and CISH was 89.7% (243/271). Two results showed significant correlation (P<0.01). (3) Paired CISH/FISH results were concordant in 14 of 15 cases. The remaining case was detected by FISH, but showed no HER2 gene amplification by CISH. (4) The gene amplification by CISH had a significantly reverse correlation with ER and PR expression (P<0.01).

CONCLUSIONSThe results of HER2 gene amplification detected by CISH have high concordance with the results detectd by IHC and FISH. CISH is a novel technique for detecting HER2 gene amplification.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization ; methods ; Prospective Studies ; Receptor, ErbB-2 ; genetics ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies

OBJECTIVETo study the segregation of two novel RHD alleles in Chinese pedigrees.

METHODSThe Rh antigens of the samples were identified by using monoclonal antibodies. The 10 exons of the RHD gene for the 2 probands and their family members were amplified separately and sequenced. The parents of proband 2 were analyzed by sequence specific primer-polymerase chain reaction (SSP-PCR).

RESULTSThe two probands were RhD negative and the RHD was D/d type. After alignment with the nucleotide sequence in GenBank, a deletion of nucleotide C at position 78 in exon 1 of proband 1 was detected, and her sister also had the deletion, which was confirmed by sequencing. The sequencing results of proband 2 showed a 10 nucleotide deletion in exon 8 as well as a RHD 520 G to A mutation in exon 4. The results of SSP-PCR and sequencing showed that the proband's mother also carried RHD 520 G to A and RHD 1080 del 10 mutation, which was transmitted to proband 2. The sequences of the novel alleles have been submitted to GenBank (accession No. GQ477180 and GU362076).

CONCLUSIONThe two novel RHD alleles, RHD 78delC and RHD 520 G to A+1080 del 10, were both pseudo genes and stably transmitted.

Adult ; Alleles ; Base Sequence ; Exons ; Female ; Genotype ; Humans ; Male ; Pedigree ; Rh-Hr Blood-Group System ; genetics ; Young Adult

Objective To study the effects of Twist gene silencing on migration and invasion of human triple-nega-tive breast cancer cell Hs578T and its potential mechanisms. Methods The lentiviral vectors with Twist gene-targe-ted specific shRNA and the negative control were constructed,then transfected into the human triple-negative breast cancer cell line Hs578T to establish the stable cell lines of Twist gene silencing. The blank group(blank),negative control group(shNC) and Twist gene silencing group(shTwist) were set up.After screening by puromycin,the fluo-rescence expression and the infection efficiency of each group cells were observed by inverted fluorescence micro-scope. The expression of Twist mRNA and protein level was detected by quantitative real-time PCR and Western blot.Transwell migration and invasion experiments were performed to measure cell migration and invasion abilities, re-spectively. Western blot was used to detect the levels of p-AKT, AKT, p-ERK1/2 and ERK1/2 proteins. Results Twist gene was successfully knockdown in human triple-negative breast cancer cell line Hs578T. Compared with the blank group and shNC group cells,the cell migration and invasion ability of the shTwist group cells were decreased significantly(P<0.05,P<0.05),and the expression of Twist downstream p-AKT and p-ERK1/2 proteins were al-so reduced notably in the shTwist group cells(P<0.05,P<0.05). Conclusions Twist promotes cell migration and invasion via AKT/ERK signaling axis in human triple-negative breast cancer cell line Hs578T.

Apolipoprotein AI (apo AI), the major protein component of human high-density lipoprotein (HDL), is a single-chain polypeptide of 243 amino acids. Several epidemiological studies have shown that the plasma concentrations of HDL has the role of reverse cholesterol transport (RCT) and inversely correlated with the incidence of coronary artery disease. Because apo AI lacks post-translational modifications, it is convenient to express human apo AI in Escherichia coli expression system. However, there is a poor stability of the mRNA and the apo AI protein in E. coli, it is difficult to express mature apo AI in recombinant bacteria, moreover, even as a fusion protein, apo AI is still sensitive to degradation and can not be cleaved efficiently from the fusion tags. In contrast, proapolipoprotein AI (proapo AI, having an additional polypeptide containing the amino acids Arg-His-Phe-Trp-Gln-Gln at the amino-teminal of the mature protein) proved stable and undegraded in Escherichia coli, and therefore, in this research, an expression system of E. coli including a plasmid of P(R)P(L) tandem promoter was adapted to produce proapo AI. Furthermore, site-directed mutagenesis of the proapo AI cDNA was performed to generate a Clu8Asp mutation in the amino-terminal sequence of proapo AI which created an acid labile Asp-Pro peptide bond between amino acid 8 and 9, and permitted specific chemical cleavage to remove pro-peptide. After inducing with a shift of temperature, yields of recombinant proapo AI achieved about 40% of total cell protein and the recombinant proapo AI expressed proved as a form of inclusion body in cells, so protein need to renature. First of all, the protein was dissolved in buffer with denaturant, and renaturation was carried out on a hydrophobic interaction column (Phenyl Sepharose), ion-exchange chromatography and gel-filtration chromatography were then used to further purify the protein. The purified recombinant apo AI was detected by a set of tests including Western-blotting, Circular dichroism spectra and lipid-binding test, the results shown that recombinant apo AI has similar structural and lipid-binding properties identical to those of native plasma apo AI, which facilitates further research and application.
Apolipoprotein A-I ; biosynthesis ; genetics ; Chromatography, Ion Exchange ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Protein Precursors ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification

OBJECTIVESTo investigate the prevalence of oncogenic type of human papillomavirus (HPV) infection and identify the high risk population for conducting immuno/chemoprevention of cervical cancer.

METHODSAll married women aged 30 to 50 with no history of hysterectomy, pelvic radiation and non-pregnant from certain villages of Xiangyuan and Yangcheng County were invited. This study was conducted through two phases. In phase one, subjects sampled the vaginal secretions using the collectors after signing the informed consent. And physicians sampled exfoliated cells from cervix in the phase two. All the specimens were tested with the Hybrid Capture 2 test. The data was managed and analyzed by VFP and SPSS software.

RESULTSThere were 9,683 women participated in this study. Local women welcomed this study and population compliance rate was 75.4%. In tested population, we found 2,666 subjects of HPV DNA positive and HPV prevalence was 27.5%. The rates of different age group were 24.5% (30-34 yrs), 27.4% (35-39 yrs), 28.2% (40-44 yrs), 27.4% (45-50 yrs) respectively and had no significant differences (P = 0.604). The rates were slightly increased with the higher education level and had no differences (P = 0.106). The rate in mountain areas was higher than that in half-mountain areas (P = 0.001).

CONCLUSIONSThe prevalence of HPV infection is indeed high in this region. Local women and health professionals welcome the activities of cervical cancer screening and prevention. It is an emergent task to improve their sanitary condition and prevent them from cervical cancer in these women. A women health cohort is established successfully among high HPV exposed women in rural China. The extensive biologic specimen repository has been successfully established to simultaneously study the etiology, early detection, and immuno/chemoprevention of cervical cancer.

Adult ; China ; epidemiology ; DNA, Viral ; analysis ; Female ; Humans ; Mass Screening ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; epidemiology ; prevention & control ; Prevalence ; Tumor Virus Infections ; epidemiology ; prevention & control ; Uterine Cervical Neoplasms ; epidemiology ; prevention & control ; virology ; Vaginal Smears