Objective To investigate the features of white matter impairment and its relationship with cognition in patients with amnestic mild cognitive impairment (aMCI).Methods Eighty-three cases of aMCI and 85 normal aging volunteers were scanned with diffusion tensor imaging (DTI) using MR system.All subjects completed the neuropsychological battery.We analyzed the differences between two groups using tract-based spatial statistics and the association between regions in difference and cognition using correlation analysis.Results There were significant differences between aMCI and normal control in the neuropsychological battery including the Mini-Mental State Examination(26.2 ± 2.6 vs 28.3 ± 1.3,F =43.224,P =0.000),Mattis Dementia Rating Scale-2 (131.4 ± 6.9 vs 138.0 ± 3.5,F =62.308,P =0.000),Auditory Verbal Learning Test-delayed recall(2.4 ± 1.6 vs 7.5 ± 2.0,F =324.018,P =0.000),Boston Naming Test(8.7 ± 1.4 vs 9.2 ± 1.0,F =6.821,P =0.010),Rey-Osterrich Complex Figure Test (12.1 ± 7.3 vs 18.5 ± 6.1,F =40.674,P =0.000),Symbol Digit Modulation Test (30.0 ± 10.1 vs 38.6 ± 9.8,F =30.786,P =0.000),Trail-Making Test Part B ((256.8 ± 124.5) s vs (178.1 ± 59.0) s,F =27.601,P =0.000).Significantly higher diffusivity indexes and radial diffusivity were also found in aMCI subjects compared to healthy elders in the parahippocampal,superior longitudinal fasciculus,inferior longitudinal fasciculus,superior fronto-occipital fasciculus,inferior fronto-occipital fasciculus,unciform fasciculus,corticospinal tract,corpus callosum,cingulum,corona radiate.We also found that axial diffusivity was significantly increased in the parahippocampal,superior longitudinal fasciculus,inferior longitudinal fasciculus,superior fronto-occipital fasciculus,inferior fronto-occipital fasciculus,unciform fasciculus,corticospinal tract and corpus callosum,whereas fractional anisotropy changes were not observed in aMCI.Diffusivity indexes values in bilateral frontal lobe (left r =0.67 ; right r =0.70),left cingulum (r =0.63),parietal white matter (r =0.69) and radial diffusivity values in left parietal (r =0.68) were significantly related to Trail Making Test A among aMCI (all P ＜ 0.05).Conclusions In aMCI patients,there was a wide range of white matter damage,with no brain region-specific.Executive function deficit was related to the white matter impairment in bilateral frontal lobe,left cingulate and parietal lobe.The specificity and sensitivity of four DTI parameters fordetecting white matter lesions are variant.Trial registration Clinical Research Center of Jiangsu Province (BL2013025)

Objective: To analyze the relationship between hemoglobin ( Hb ) and serum uric acid ( SUA ), so as to provide basis for preventing hyperuricemia ( HUA ) . Methods: As the research subjects, people who underwent physical examination in Zhejiang Provincial People's Hospital from January 1, 2017 to October 31, 2020 for 4 years in a row and who were non-HUA in 2017 were selected. The physical examination data were collected, including body mass index, blood pressure, blood routine, blood biochemical tests, etc. The subjects grouped by quartiles of Hb level in 2017. The occurrence of SUA elevation ( SUA increased ≥60 μmol/L from baseline ) , significantly SUA elevation ( SUA increased ≥120 μmol/L from baseline ), HUA ( SUA>420 μmol/L ) and severe HUA ( SUA ≥480 μmol/L ) in the next 3 years were taken as end events. The incidence, occurrence time and risk of end events in different Hb groups were analyzed. Results: A total of 4 073 subjects were selected and divided into 4 groups according to the Hb level from low to high, with 969 subjects in group A, 907 subjects in group B, 1 109 subjects in group C and 1 088 subjects in group D. SUA elevation was in 745 patients ( 18.29% ); significantly SUA elevation was in 105 patients ( 2.58% ); HUA was in 514 patients ( 12.62% ); severe HUA was in 94 patients ( 2.31% ). The incidence of SUA elevation and significantly SUA elevation showed a decreasing trend with the increase of Hb level ( P<0.05 ). The occurrence time of SUA elevation in group A to D was 2.788, 2.817, 2.860 and 2.814 years, and the difference was statistically significant ( P<0.05 ). There were no statistically significant differences in the occurrence time of other end events ( P>0.05 ). The multivariate Cox proportional hazards regression analysis showed that compared with group A, other Hb groups had lower risk ( HR=0.498-0.776, 95%CI:0.253-0.981 ) of SUA elevation, significantly SUA elevation and severe HUA after adjusting for gender, age, ALT, Scr, body mass index, etc. Conclusions With the increase of Hb level, the incidence of SUA elevation may decrease and the occurrence time is prolonged. Compared with the lowest Hb group, the higher Hb groups had lower risk of SUA elevation, significantly SUA elevation and severe HUA.

OBJECTIVE CYP2D is one of the most abundant subfamily of CYPs in the brain, especially in the cerebellum. Brain CYP2D is responsible for the metabolism of endogenous neurotransmitters such as tyramine and serotonin. Our previous studies have shown brain CYP2D can be regulated by exogenous and endogenous substances with tissue- specificity. The purpose of this study is to examine the effects of cerebral CYP2D on the mice behavior and the regulatory mechanism of brain CYP2D by growth hormone. METHODS Mice received the stereotaxic injection with CYP2D inhibitor quinine in deep cerebellar nuclei of cerebellum. The animals were tested with rotarod apparatus, balance beam, water maze, elevated plus maze and open field. The changes in CYP2D22, PPARαand PPARγ in brain regions and liver were assayed in male growth hormone receptor knockout mice, SH-SY5Y cells and HepG2 cells. RESULTS The inhibition of cerebellum CYP2D significantly affected the spatial learning and exploring ability of mice. Compared with WT mice, CYP2D expression was lower in brain regions from GHR(-/- ) male mice; however, hepatic CYP2D level was similar. Pulsatile GH decreased PPARα mRNA level, and increased mRNA levels of CYP2D6 and PPARα in SH- SY5Y cells. In HepG2 cells, pulsatile GH resulted in decreases in PPARα and PPARγ mRNA levels, but not CYP2D6. PPARα inhibitor induced CYP2D6 mRNA and protein by 1.32-fold and 1.43-fold in SH-SY5Y cells. PPARγ inhibitor decreased CYP2D6 mRNA and protein by 74.76% and 40.93%. PPARα agonist decreased the level of CYP2D22 mRNA in liver and cerebellum, while PPARγ agonist rosiglitazone resulted in diametrically increases. The luciferase assay showed that PPARγ actived the CYP2D6 gene promoter while PPARα inhibited its function. Pulsatile GH declined the binding of PPARα with CYP2D6 promoter by 40%, promoted the binding of PPARγ with CYP2D6 promoter by approximate 60%. The levels of brain and liver PPARα expression in male GHR(-/- ) mice is obviously higher than those in WT mice. The level of PPARγ in male GHR(-/- ) mice was decreased in the frontal cortex and hippocampus, while remained stable in the cerebellum and striatum; meanwhile, PPARγ was increased in the liver. CONCLUSION Brain CYP2D may be involved in learning and memory functions of central system. Masculine GH secretion altered the PPARs expression and the binding of PPARs to CYP2D promoter, leading to the elevated brain CYP2D in a tissue- specific manner. Growth hormone may specifically alter the metabolic and synthetic of important endogenous substances in the central nervous system (such as serotonin) through the specific regulation of brain CYP2D expression.

OBJECTIVETo study role of dental follicle in tooth root development.

METHODSSixteen mandibular first molar dental germs from eight five-day postnatal Balb/c mice were divided into two groups randomly. Dental follicle of germs in one group was undetached and that of another group was removed. Subsequently, each of the germs was separately transplanted to back-muscles of adult nude mice. At seventh and fourteenth day after transplanting, the germs were collected, fixed, demineralized, dehydrated, and embedded in wax in sequence. Serial sections of 5 microm thick were made following the routine methods, stained with haematoxylin-eosin dying solution, and observed under a light microscope.

RESULTSAll implantations were located in the back-muscles with abundant capillary vasculature. Under microscope, although all tooth germs could further develop after grafting, tooth germs without dental follicle developed slowly with small size and low calcification compared to those with dental follicle. Although position of Hertwig's epithelial root sheath of all germs seemed no changing, roots of the group with dental follicle could further develop and the roots develop toward the apical direction; this tendency couldn't be seen in the germs of another group. Inflammatory cells could be seen in and out of the pulp cavity of the two groups at 7th day after grafting, while no obvious inflammatory cell was observed at 14th day after grafting.

CONCLUSIONDental follicle play an important role in tooth root development. It probably can lead tooth root to develop in normal direction.

Animals ; Dental Pulp Cavity ; Dental Sac ; Enamel Organ ; Mice ; Mice, Nude ; Molar ; Odontogenesis ; Tooth ; Tooth Germ ; Tooth Root

To explore the potential of the anti-sense nucleic acid of CyclinD1 in lung cancer therapy, the expression vector containing the anti-sense nucleic acid of CyclinD1 was constructed and named pcDNA3.1-CyclinD1. The A549 cells were transfected with pcDNA3.1-CyclinD1 vectors. After being screened by G418, the stable expression positive clones were obtained. MTT method and flow cytometry technique were used to detect cell proliferation and apoptosis, respectively. The results showed the transfected cells exhibited significantly increased apoptosis and inhibited cell growth, compared with negative control and empty vector groups. To investigate the mechanism for anti-sense nucleic acid of CyclinD1 inducing A549 cells apoptosis, the expression levels of retinoblastoma protein (pRb), adenovirus E2 factor-1 (E2F-1), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blot, and the results showed the expressions of these proteins were all decreased significantly in anti-sense nucleic acid of CyclinD transfected group, compared with those in negative control and empty vector groups. In a word, anti-sense nucleic acid of CyclinD1 induces the apoptosis of lung adenocarcinoma cancer cells, and the depressions of pRb, E2F-1, VEGF, MMP-2 and MMP-9 expressions may be the possible mechanism.
Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cyclin D1 ; genetics ; DNA, Antisense ; pharmacology ; Genetic Vectors ; Humans ; Lung Neoplasms ; pathology ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Recombination, Genetic ; Retinoblastoma Protein ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

Animals ; Fatty Liver ; drug therapy ; metabolism ; prevention & control ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Taurine ; therapeutic use

OBJECTIVETo clarify the role of RhoC in the growth of hepatocellular carcinoma cells and its molecular mechanism, so as to explore the molecular target of tumor cell growth.

METHODSsiRNA-RhoC plasmid was constructed and RhoC gene silencing the cell-line of hepatocellular carcinoma was setup. Cell growth was assessed by MTT assay. AgNORs staining was applied to determine cell proliferation. Plate cell clone test was conducted to examine the capacity of cell clone formation. FACS was adopted to measure the course of cell cycle and semi-quantitative RT-PCR was used to determine the expression of cell cycle proteins. In order to further determine the effect of RhoC expression on cell growth, a RhoC over-expression human hepatocellular cell line was setup by PcDNA3-RhoC plasmid transfection.

RESULTSThe inhibition rate of RhoC was 82.3%. From the fourth day of cell culture, the growth of cells in RNAi group was significantly slower than that in parental Bel7402 and negative control groups (0.41 ± 0.10 vs. 0.73 ± 0.11 and 0.71 ± 0.07 respectively, P < 0.05). AgNORs staining showed that average cell stained particles in RNAi group was significantly lower than that in parental Bel7402 and negative control(1.23 ± 0.35 vs. 3.47 ± 0.93 and 3.17 ± 0.78, P < 0.01). Plate clone formation test showed that clone formation efficiency in the RNAi group was notably lower than that in the control group [(20.33 ± 5.42)% vs. (70.58 ± 10.10)% and (69.83 ± 14.77)%, respectively, P < 0.01]. Cell cycle analysis by FACS showed that G(0)/G(1) cell percentage in the RNAi group was significantly higher than that in the control group [(73.14 ± 5.93)% vs. (57.05 ± 5.97)% and (52.99 ± 4.80)%, P < 0.05]. Compared with Bel7402 and negative control groups, the expression of following growth associated genes was significantly decreased: cyclin D1(0.45 ± 0.21 vs. 1.25 ± 0.24 and 1.12 ± 0.15, respectively, P < 0.05)and CDK4 (0.55 ± 0.08 vs. 1.18 ± 0.32 and 1.10 ± 0.29, respectively, P < 0.05); the following genes were notably increased: p16(1.07 ± 0.23 vs. 0.36 ± 0.12 and 0.35 ± 0.13, respectively, P < 0.01)and p21(0.42 ± 0.12 vs. 0.17 ± 0.06 and 0.19 ± 0.08, respectively, P < 0.05). RhoC was highly expressed in PcDNA3-RhoC transfected hepatocellular cell line. From the third day on of the cell culture, cell growth in PcDNA3-RhoC group was remarkably higher than that in the HL7702 and PcDNA3 groups (0.83 ± 0.10 vs. 0.54 ± 0.11 and 0.58 ± 0.55, respectively, P < 0.05).

CONCLUSIONSRhoC is the key molecule in promoting hepatocellular cell growth, and is a promising target for tumor cell growth controlling.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; rho GTP-Binding Proteins ; genetics ; metabolism ; rhoC GTP-Binding Protein

OBJECTIVETo study the distribution and expression of fibromodulin, decorin and biglycan in developing normal periodontal tissues, so as to understand its role in periodontal tissue formation.

METHODSThirty six BALB/c mice in different developing stages were killed and their bilateral mandibular first molars with surrounding alveolar bones and gingival tissues were taken out, Power Vision two steps immunohistochemical method with anti-fibromodulin, anti-decorin and anti-biglycan was used to detect the tissue distribution and cellular localization of fibromodulin and related proteoglycans, decorin and biglycan.

RESULTSFibromodulin was strongly expressed in the subcutaneous gingival connective tissue, periodontal ligament, mainly in gingival and periodontal fibroblasts as well as their matrices. Strong expression was also noted in the area close to the interfaces of periodontal ligament-alveolar bone and periodontal ligament-cementum. Decorin was strongly expressed in the area of gingival connective tissue, periodontal ligament and the surface of alveolar bone, while biglycan was stained evidently in gingival connective tissue throughout the period of investigation, but negative in the surface of alveolar bone and osteoblasts.

CONCLUSIONSFibromodulin may interact with decorin and biglycan to regulate the network formation of gingival connective tissues and periodontal collagen fibers, and may be involved in mineralization of the alveolar bone and cementum.

Alveolar Process ; cytology ; growth & development ; Animals ; Biglycan ; Decorin ; Extracellular Matrix Proteins ; analysis ; Fibromodulin ; Gingiva ; chemistry ; growth & development ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Osteoblasts ; chemistry ; Periodontal Ligament ; chemistry ; growth & development ; Proteoglycans ; analysis ; Tooth Germ ; chemistry

Objective To optimize the extraction technology for Yangxin Anshen Granules. Methods With yield of volatile oil as index, single factor tests were adopted to investigate effects of water, soaking time and distillation time on extraction technology of volatile oil. Using water amount, extraction time and extraction times as factors, the contents of paeoniflorin and total solid as indexes, orthogonal test was employed to optimize the extraction technology of Yangxin Anshen Granules. Results The optimical extraction technology conditions were as follows:Cinnamomi Ramulus, Saposhkoviae Radix, and Citri Sarcodactylis Fructus were extracted to get volatile oil with eight-folds amount water of herbs for 6 hours; other herbs were boiled with eight-folds amount water of herbs and extrancted for two times, 1 h each time. Conclusion This extraction process is reasonable and practical, and can well guarantee the quality of preparation.

The comparative efficacy of 2 anthelmintics (ivermectin and levamisole) against Baylisascaris transfuga migrating and encapsulated larvae was studied in mice. A total of 60 BALB/c mice inoculated each with about 1,000 embryonated B. transfuga eggs were equally divided into 6 groups (A-F) randomly. Mice of groups A and B were treated with ivermectin and levamisole, respectively, on day 3 post-infection (PI). Mice of groups A-C were killed on day 13 PI. Similarly, groups D and E were treated with ivermectin and levamisole, respectively, on day 14 PI, and all mice of groups D-F were treated on day 24 PI. The groups C and F were controls. Microexamination was conducted to count the larvae recovering from each mouse. The percentages of reduction in the number of migrating larvae recovered from group A (ivermectin) and B (levamisole) were 88.3% and 81.1%, respectively. In addition, the reduction in encapsulated larvae counts achieved by ivermectin (group D) and levamisole (group E) was 75.0% and 49.2%, respectively. The results suggested that, to a certain extent, both anthelmintics appeared to be more effective against migrating larvae than encapsulated larvae. However, in the incipient stage of infection, ivermectin may be more competent than levamisole as a larvicidal drug for B. transfuga.
Animals ; Anthelmintics/*administration & dosage ; Ascaridida Infections/*drug therapy/parasitology ; Ascaridoidea/*drug effects ; Disease Models, Animal ; Female ; Ivermectin/*administration & dosage ; Larva/drug effects ; Levamisole/*administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Rodent Diseases/drug therapy/parasitology ; Treatment Outcome