Objective To observe the depositing regions of the cerebral neurons containing A?_ 42 neurons and to study the effects of ginkgo leaf extracts and dioyridamole injection on the memory,the deposition of A?_ 42 in the brain of Alzheimer's disease(AD) model rats induced by aluminium chloride(AlCl_3).Methods The experimental AD model of rats was established by 30% AlCl_3.The rats of treatment group received ginkgo leaf extracts and dioyridamole injection for consecutive six months.Latent period and the intensity of electric shock was evaluated by a passive-avoidence test before and after administration respectively.The morphology and the number of A?_ 42 neurons of the model rats brain by light microscope observation as well as immnohistochemistry.Results (1)After treatment,latent period of the rats prolonged and the intensity of electric shock reduced as compared with AlCl_3 group.(2)The depositing regions of A?_ 42 were found mainly in pyramidal cell layer of CA_1 hippocampus formation and polymorphical cell layer of dentate gyrus,external main stratum of entorhinal region and Ⅲ,Ⅵ,Ⅴ cell layer of cortex(parietal,temporal).The color of A?_ 42 positive substance was brown or brown yellow in the cytoplasm and its dendrites or branches.Significant reducing of A?_ 42 neurons occurred in treatment group(hippocampus P0.05).Conclusion Ginkgo leaf extracts and dioyridamole injection can improve the AD model rats ability of learn and memorizing,which mechanisms are achived by preventing A? from depositing and clearing deposited A?.

Objective To explore whether respiratory tract viruses infection was involved in the pathogenesis of steroid sensitive and simple nephrotic syndrome(SSSNS).Methods Expression of respiratory tract viruses gene and antigen in peripheral blood mononuclear cells(PBMC) and production of viruses antibody in sera from 26 children with steroid-sensitive and simple nephrotic syndrome were evaluated by reverse transcription polymerase chain reaction(RT-PCR),APAAP and ELISA.Results In compared in active phase with remission group of SSSNS and normal age-matched controls,the positive rate of viruses of active phase group was significantly increased(P

Aim: To observe the effects of sarsasapogenin ( SAR) on osteoblasts and osteoclasts cultured in vitro. Methods: Colonal murine calvarial osteoblast-like cell line MC3T3-E1 cells were cultured in vitro. MTT,p-nitropheneye phosphate and tinctorial method of alizarin Bordeaux were used to investigate the effects of SAR on the proliferation, ALP expression, and mineralization tuberculation of MC3T3-E1 cells. Mature osteoclasts were i-solated from the long bone of one-day rat. Meanwhile, marrow cells of mouse bone were cultured with induction of 1,25( OH)_2VitD_3. During the culturing of osteoclasts or marrow cells, SAR of different concentrations was added into the medium. The number of osteoclasts was recognized as tartrate resistant acid phosphatase( TRAP) ( +) multinucleate cells and the resorption lacuna on bone slice were examined with toluidine blue staining. Results: Comparing with the control group, SAR (0.01, 0. 1, 1μg/mL) significanthy increased the proliferation of MC3T3-E1 cells (P <0. 05, P <0. 01). There was no significant difference in the expression of ALP in early pro-liferating MC3T3-E1 cells exposed to SAR of 0.01,0. 1, 1μg/mL, but in the differentiation phase MC3T3-E1 cells, SAR improved ALP activity very significantly if compared with the control group, of which SAR of 1 μg/mL had the most promotion effect(P <0. 01). In addition, compared to the control group, there were, to various ex-tents, increased in the number of mineral nodes in MC3T3-E1 cells after 15day incubation with SAR of different conentrations. Furthermore, no obvious effects of 0.01-1μg/mL SAR on mature osteoclast were observed. But typical osteoclasts were formed when marrow cells were cultured with the induction of 1,25(OH)_2D_3 in medium for 7 days while little or no osteoclasts were induced from marrow cells in the presence of SAR. Conclusion: The results suggest that SAR can effectively promote the proliferation, differentiation and mineralization of osteoblasts cultured in vitro. Besides, SAR can inhibit the generation of osteoclasts from marrow cells.

Objective To investigate the relationship between the expression of glucose regulated protein 78 (GRP78) in the placental trophoblast cells and the pathogenesis of gestational diabetes mellitus (GDM).Methods All the patients were recruited from Qingdao Municipal Hospital from May 2013 to May 2014.Among them, fifty women with GDM were assigned to the GDM group, and fifty healthy women were defined as the control group.All of them received cesarean section because of breech presentation, contracted pelvis, scarred uterus or on mother's demand.Real-time PCR was conducted to analyze the expression of GRP78 mRNA in the trophoblasts.Immunohistochemistry was performed to detect the localization of GRP78 protein in the placentasl trophoblast cells.Results (1) GRP78 mRNA expressed in the cytoplasm of trophoblasts of both the GDM group and the control group.The GRP78 mRNA levels in the GDM group and the control group were 15.6±0.4 and 6.0±0.7, respectively.The relative expression level of GRP78 mRNA in the GDM group was 2.6 times of that in the control group, with statistically significant difference (P＜0.01).(2) The expression of GRP78 protein was found in the cytoplasm of the trophoblasts of the GDM group.It showed in deep, light brown or yellow after staining, according to the expression degree.The expression of GRP78 protein was also found in the cytoplasm of the trophoblasts of the control group, but it mainly showed yellow color (38/50).The strong positive rate of GRP78 protein in the GDM group (96％, 48/50) was higher than that in the control group (22％, 11/50;P＜0.01).Conclusion The expression of GRP78 increased in the placental trophoblast cells of GDM patients.It might suggest that GRP78 had some effect on the pathogenesis of GDM.

Objective To investigate the effects of the transient receptor potential V6 (TRPV6) gene silencing on the proliferation and apoptosis of trophoblasts HTR-8/SVneo cells.Methods siRNA sequences targeting the TRPV6 gene were constructed and then transfected into HTR-8/SVneo cells mediated by liposome.The cells were divided three groups,including blank control (add the reagent of transfenction),negative control groups (transfecting nonspecific siRNA) and experimental groups (transfecting TRPV6-siRNA).Those cells in every group were collected at 24,48,72 hours after transfecting.The expression levels of TRPV6 mRNA were detected by reverse transcription (RT) PCR at different times after transfecting.The effects of siRNA on the proliferation and apoptosis of the cells were assayed by methyl thiagolyl tetragolium (MTT) and flow cytometry at different times after transfecting.Results siRNA TRPV6 transfection could inhibit the expression of TRPV6 mRNA in the HTR-8/SVneo cells.The expression was decreased with the extension of time,by 0.72 ± 0.02,0.54 ± 0.02 and 0.29 ± 0.01 after 12,48 and 72 hours of siRNA transfection as compared with the blank control and the negative control groups (P ＜0.01).The rates of proliferation inhibition were (19.29 ± 1.23) ％,(32.12 ± 1.35) ％ and (46.51 ±1.42) ％ at 24,48 and 72 hours respectively when compared with the blank control (2.12 ± 0.03)％,(2.42 ± 0.02) ％,(3.13 ± 0.04) ％ and the negative control groups (2.37 ± 0.01) ％,(2.61 ± 0.05) ％,(2.93 ± 0.03) ％ (P ＜ 0.01).The apoptosis rates of HTR-8/SVneo cells was 16.21％ at 48 hours after transfected with siRNA TRPV6,which were significantly higher than 3.27％ in the blank control and 5.34％ in the negative control groups (P ＜ 0.05).Conclusion Silenceing of TRPV6 genen could inhibit the proliferation and increase the apoptosis of extravillous trophoblas of human placenta.

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

Objective To evaluate the effectiveness of mouth care combined with intestinal and endoscopic working channel washing for ERCP related cholangitis.Methods A total of 573 patients diagnosed as having obstructive jaundice were randomized into three groups,190 cases in the control group,192 in the saline group and 191 in the amikacin group.Clinical and laboratory data were collected before ERCP and ERCP related cholangitis were recorded.Results There were no significant differences among the three groups in sex,age,the level of obstruction,the category of obstruction,total bilirubin or WBC counting.The incidences of ERCP related cholangitis were 21.1％ (40/190),13.5％ (26/192) and 4.7％ (9/191)in control group,saline group and amikacin group respectively,which was significantly different (x2 =22.409,P =0.000 ).The incidences of ERCP related cholangitis were 19.5％ ( 65/333 ) and 4.2％(10/240) in patients diagnosed as having hilar duct obstruction and low positioned biliary obstruction respectively (x2 =27.175,P =0.000).There was no significant difference in ERCP related cholangitis between benign and malignant biliary obstruction.Subgroup of hilar duct obstruction showed the incidences of ERCP related cholangitis were 29.7％ (33/111 ),20.5％ (24/117)and 7.6％ (8/105)in the control group,the saline group and the amikacin group,respectively (x2 =16.905,P =0.000).Conclusion The incidence of ERCP related cholangitis is relatively higher in patients with hilar duct obstruction.Mouth care combined with intestinal and endoscopic working channel washing can effectively reduce the incidence of ERCP related cholangitis,especially in the amikacin group.

Objective To investigate the effects and mechanism of retinoie acid on B16 marina mel-anoma cells and human melanocytes in vitro. Methods B16F10 murine melanoma cells and human mela-noeytes were cultured in culture medium which contains different concentration of components, including retinoic acid. Using reverse transcription-polymerase chain reaction (RT-PCR) mRNA expression of the tyrosinase was detected. Tyrosinase activity, melanin content and cell proliferation rate were also deter-mined. Results Retinoieacid exhibited an inhibitory effect on the expression of tyrosinase mRNA. As the concentration of retinoic acid was 100 μmol/L, treating for 72 h, the expression of tyrosinase mRNA de-creased 30.13 %, retinoic acid exhibited an inhibitory effect on tyrosinase activity and melanin production at high concentration (>500 μmol/L), and it could promote the cell proliferation. Retinoic acid and hy-droqninone could be cooperative at high concentration (1 000 μmol/L), and enhanced the down regulation of tyrosinase activity and melanin content. Retinoic acid could also mitigate the inhibitory effect of hydro-quinone on cell proliferation, so as to protect the cells from injury. Hydroquinone had no effect on tyrosi-nase gene expression at mRNA level. Conclusion Retinoic acid inhibits the synthesis of melanin by the genetic regulation at mRNA level.

Humans ; Sphenoid Bone ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed

Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.