Objective To explore the effect of icariin on stimulating Smad4 mRNA level in MC3T3-E1 cells. Methods MC3T3-E1 cells were treated by 0 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml icariin respectively. MTT method and population diploid time were applied to observe the cell proliferation. The cell ALP level was measured by atomic absorptiometry and the type Ⅰ collagen was observed by immunohisto-chemistry. RT-PCR was used to find the Smad4 mRNA level in the MC3T3-E1 cells. Results Under the inversion phase contrast microscopy, MC3T3-E1 cells were oligodendrocytes-shaped rich with cell organs. The cells density in 10 ng/ml, 20 ng/ml were higher than those in 0 ng/ml,40 ng/ml. Cell growth curve in 10 ng/ml was higher than other 3 groups and the 20ng/ml was higher than 0 ng/ml, 40 ng/ml groups( P

Objective To explore the diagnostic value of LUNX gene to marrow micrometastases of lung cancer.Methods To detect LUNX mRNA of marrow samples of 51 patients of lung cancer,4 patients of breast cancer,6 patients of lymphadenoma,3 patients of liver cancer and 22 patients of benign disease by real-time RT-PCR.Results The positive detection rate and meso-copies of lung cancer were 58.8% (30/51) and 35copies/ml respectively.The positive detection rate and meso-copies of other diseases were all 0,The positive detection rate and meso-copies of lung cancer was significantly higher than that of patients with other diseases (x~2=11.12,U_c=3.7329,P

Objective: To establish a method for the determination of glucosamine hydrochloride content in aminoglycine enteric coated tablets by high-performance liquid chromatography with differential refractive detector.
Methods: Waters XBridge Amide (4.6 mm×250 mm, 3.5 μm) column was used. The mobile phase was 80% acetonitrile solution (1 000 mL containing 1 mL ammonia) at the flow rate of 1.2 mL·min^-1. Column temperature was set on 40 ℃; Injecting volume was 20 μL with differential refraction detector (RID) detection.
Results: The detection limit of glucosamine hydrochloride was 0.03 mg·mL^-1, and the limit of quantification was 0.125 mg·mL^-1. It showed a good linear relationship in the mass concentration range of 0.72-7.56 mg·mL^-1. The average recovery was 99.4% (RSD=0.85%,n=9). The content of glucosamine hydrochloride in 7 batches of aminosamine enteric-coated tablets was measured in the range of 93%-107%.
Conclusion: The proposed method is fast, accurate, specific and durable, and can be used for the determination of glucosamine hydrochloride, which provides a reference for effectively controlling the quality of aminoxymetacin enteric-coated tablets.

To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Escherichia coli ; genetics ; metabolism ; Influenza A virus ; genetics ; metabolism ; Influenza B virus ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Ribonucleases ; chemistry ; Virion ; genetics ; metabolism

ABSTRACT:Objective To study the platelet changes in patients with unstable angina with different blood glucose ,and their related biochemical index changes ,and their relationship with global registry of acute coronary events (GRACE) score .Methods For this clinical study ,we enrolled 82 patients diagnosed with unstable angina , 47 of whom were male and 35 were female .Upon admission ,their random blood glucose was tested .According to different blood glucose values ,they were divided into normal blood glucose group (<6 .1 mmol/L) and high blood glucose (≥ 6 .1 mmol/L ) group . The following clinical data were compared between the two groups :age , hypertension ,diabetes ,smoking history ,and BMI .We detected EF (% ) ,HBA1C ,glucose ,LDL‐C ,HDL‐C ,TG , LPA ,CREA ,UA ,hsCRP ,BNP ,CKMB ,CTNI ,D‐Dimer ,and GRACE risk scores .We compared the platelet test results :PLT ,P‐LCR ,PDW ,and MPV .We also detected the relationship of MPV with hsCRP ,D‐Dimers and GRACE risk scores .Results MPV ,hsCRP ,and GRACE risk score differed significantly between normal blood glucose group and high blood glucose group (P<0 .05) .In the latter group ,MPV had significant correlation with hsCRP ,D‐Dimers and GRACE risk score ( r=0 .28 , r=0 .41 , r=0 .56 , P<0 .05) .Conclusion Hyperglycemia in patients with unstable angina causes the increase of MPV , change of the inflammatory marker hsCRP , and increase of clinical GRACE risk score .Abnormal MPV may predict the increased risk of unstable angina in patients with hyperglycemia upon hospitalization .

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

AIM:To point the susceptible gene in Avellino corneal dystrophy family with autosomal dominant inheritance.
●METHODS: Genomic DNA was extracted from the peripheral blood samples of all individuals of the pedigree. Several microsatellite makers were selected for gene scan in the hot regions of mutation. Linkage analysis was carried out using a Linkage software package. The haplotype data were processed using Cyrillic software to define the region of the disease gene.
●RESULTS: ln our pedigree, significant evidence of linkage was obtained at marker D5S396 and D5S393 [LOD score (Z)=3. 01, recombination fraction (θ)=0. 00]. The haplotype analysis of our pedigree was located between the microsatellite markers D5S808 and D5S638.
●CONCLUSION:The pathogenic gene of the Avellino corneal dystrophy pedigree is traced to a 11. 2 cM region in the chromosome 5q.

Objective To observe the effects of levodopa on verbal and spatial working memory in elderly patients with Parkinson's disease (PD).Methods The modified Smith working memory software were applied to study the verbal and spatial working memory in 32 PD patients before and 8 weeks after treatment with levodopa and benserazide hydrochloride tablets.The results of PD patients were compared with those in 32 normal controls matched for age,sex and education.Results The correct rates of phonological verbal and coordinate visuospatial working memory were significantly lower in the PD patients without treatment than in the normal controls (both P＜0.01).The correct rates of semantic verbal and categorical visuospatial working memory had no significant differences between the PD patients without treatment and the normal controls (both P＞0.05).The correct rates of phonological verbal working memory and coordinate visuospatial working memory were significantly higher in the PD patients after administration of levodopa and benserazide tablets than in PD patients without treatment [(88.6±6.5)％ vs.(82.1 ± 6.6)％,(84.2±8.8)％ vs.(75.2±8.9)％,respectively,both P＜0.01],but there were no significant differences in the correct rates of semantic verbal working memory and categorical visuospatial working memory between the above two groups (both P＞0.05).The correct rates of phonological verbal,semantic verbal,coordinate visuospatial and categorical visuospatial working memory were negatively correlated with the course of disease,and had no correlations with age and the degree of education.Conclusions Levodopa can improve the functions of phonological verbal working memory and the coordinate visuospatial working memory,but has no effect on the semantic verbal working memory and the categorical visuospatial working memory.It is postulated that dopamine replacement therapy may have double dissociable effects on the different working memories.

Boronic acids and their derivatives have been widely used in carbohydrate-sensitive materials because they can selectively bind 1,2-and 1,3-diol compounds, including sugars, to form cyclic boronate esters. In this work, pheylboronic acid ( PBA) moieties were grafted onto the backbone of poly( acrylic acid) ( PAA) through the condensation reaction between aminopheyl-boronic acid and carboxylic acid group of PAA in the presence of EDC/NHS, designed as PAA-PBA. Then the resulting PAA-PBA were assembled with poly ( ethyleneimine) ( PEI) to form PAA-PBA multilayer films. The sensing performance of the PEI/PAA-PBA film to carbohydrate (> 50 μg/mL ) , including glucose, fructose, mannose and galactose, has been investigated by combination of the complementary techniques of quartz crystal microbalance ( QCM ) and atomic force microscopy ( AFM) . It was demonstrated that the multilayer showed higher sensitivity to fructose than glucose, mannose and galactose. The interferences of ascorbic acid, uric acid and dopamine to the recognition of glucose can be avoided and the multilayer sensor with excellent long term stability can be recycled by changing pH value of buffer solutions. This system may be potential in realization of high selectivity and high sensitivity sensing system for probing carbohydrate.

Objective To characterize emotional memory impairment in untreated patients with Parkinson' s disease (PD) in the early stages of the disease. Methods Emotional memory tasks using standardized sets of emotional pictures including positive, neutral and negative valence were tested in 33 untreated patients with PD in the early stages and 31 healthy controls matched with age, sex, and education.Results Compared with the healthy adults (13.4 ± 1.4), PD patients had significant loss of negative valence picture memorizing ( 8. 9 ± 1.0; t = - 14. 87, P ＜ 0. 01 ).There was no significant difference between PD patients and normal controls in positive ( 11.8 ± 1.0 vs 12. 4 ± 2. 2 , t = - 0. 95 ) and neutral (7.9 ± 1.4 vs 8.2 ± 1.3, t = - 0. 89) valence picture memorizing ( both P ＞ 0. 05 ). ConclusionThe emotional memory for negative valence pictures is impaired but the emotional memory for positive and neutral is relatively spared in early PD patients without treatment.