Objective To study the condition and method of hydrodynamics-based transgene(HDT) in rat fatty liver. Methods Inject different dosages and concentrations of green fluorescent protein plasmid pEGFP-C1 at different speeds, then collect 4 rats' liver leaves at different time points after injection and prepare their frozen section, finally observe and quantify the GFP expression with fluo rescence microscope at 488 nm excitation wavelength. Results Plasmid pEGFP-C1 concentration 33mg/L, injection speed 2ml/s, injection volume 8.5%of rat body weight, injected plasmid. After 6 hours of injection, GFP-positive cells rate of pedicel leaf is about 18%, left leaf about 14%, middle leaf about 12.5%;R3ight leaf about 10% and tail leaf about 8%. GFP begin to gradually reduce since 24 hours, until 72 hours almost no GFP-positive cells were checked in all liver leaves. Conclusion Hydrodynamics-based transgene can be applied to rat fatty liver, the appropriate conditions of this method are 33mg/L plasmid concentration, 8.5% rat avoirdupois, 2ml/s injection speed, and the suitable time to observe the proportion of GFP-positive cells is 6-24 hours after gene injection.

Objective To evaluate the diagnostic value of 64-row VCT coronary angiography(CTA) in diagnosing coronary artery disease.Methods 64-row VCT coronary angiography(CTA) and cathter coronary angiography(CAG) were performed in 173 patients with suspected coronary artery disease.All images were analysed at GE AW4.3 workstation,the constructed images included maximum intensity projection(MIP),mlti-planar reconstruction(MPR) and volume rendering(VR).The results were compared with that of CAG.Results 746 segments of coronary artery with diameter ＞1.5 mm were showed by CTA in 173 patients.The sensitivity,specificity,positive predictive value and negative predictive value of the CTA in detecting coronary arterial stenosis were 94.15%(193/203),95.90% and 89.77% and 97.90% respectively.Statistical analysis using a 2-related χ~2 test showed that there were no obvious differences in diagnosing coronary arterial stenosis between CTA and CAG(χ~2=1.58,P＞0.05) and in evaluating the stenosis degree of coronary artery(Kappa=0.890,P＜0.001).Conclusion 64-row VCT has important clinical value in screening coronary arterial disease and in following-up post operational effectiveness of coronary stent implantation and vascular bypass.

Aged ; Humans ; Male ; Middle Aged ; Receptors, Interleukin-2 ; blood ; Silicosis ; blood ; classification ; T-Lymphocyte Subsets ; metabolism

Objective To study the genomic molecular organization and genotype of human astro-virus infected infants in Shanghai of China. Methods Based on the published genomic sequence of HAstV (GenBank), the whole genome of one isolate human astrovirus was sequenced by specific primers. The PCR-products were cloned to pMD18-T vector and sequenced, phylogenetic tree was constructed by the Neighbor-Joining method with MEGA 4 software. Results The genome of HAstV-SH is 6807 hp, contains three ORFs: ORF1a and 1b encode the non-structural protein (from 83 nt to 4372 nt), ORF2 encodes the structural protein (from 4364 nt to 6727 nt). Compared with the ORF2 gene of those eight astrovirus sero-types in GenBank, revealed that the highest homology is with genotype 1 (97%). Homology with other gen-otypes ranged between 63% and 70%. Conclusion HAstV-SH belonged to genotype 1 and closely clus-tered with a strain of Japan (AB009985).

OBJECTIVETo observe the effect of Chinese herbal extract HuNan A-1 (HNA-1) on the thymic output function in Simian immunodeficiency virus (SIV) chronically infected rhesus macaques.

METHODSEight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months, and then they were randomly divided into the treatment group and the control group, 4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage, once daily for 2 successive months, while those in the control group were administered with equal volume of normal saline by gastrogavage, once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts, as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles (TREC) were detected using real-time fluorescent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue, CD4 counts, naive CD counts, and TREC were analyzed.

RESULTSThere was no statistical difference in body weight, viral loads, absolute CD ratios between the two groups after treatment (P > 0.05). The altered TREC multiple showed an obvious decreasing tendency in the control group, while it showed an increasing tendency in the treatment group (P < 0.05). In both groups, destroyed structures of the thymus tissue could be seen, filled with pink unstructured material. Increased connective tissues, lowered connective cell density, and confused arrangement could also be seen in the two groups, with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum (r = 0.926, P = 0.001). Naive CD4 counts were positively correlated with CD4 counts (r = 0.961, P = 0.005).

CONCLUSIONSTREC content determination, as a marker of newly thymic emigrants, could be taken as a testing method for evaluating the thymic output function. Besides, HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents, naive CD4 counts, and pathologies of thymus tissues, especially in late infection stage.

Animals ; CD4 Lymphocyte Count ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Macaca mulatta ; Plant Extracts ; pharmacology ; Random Allocation ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; Simian Immunodeficiency Virus ; Thymus Gland ; drug effects ; Viral Load

To study the ecological distribution and diversity of endophytic fungi associated with Ferula of medicinal plants in Xinjiang. The endophytic fungi were isolated from roots, stems and leaves of Ferula by microbiology research methods and technology. The endophytic fungi were identified using ITS rDNA sequence analysis and morphology analysis. The composition, diversity and preference of endophytic fungal community were analyzed with Shannon-Wiener biodiversity index (H') and Sorensen coefficient (Cs). A total of 337 strains endophytic fungi were isolated and classified into 38 genera, Alternaria, Aureobasidium and Fusarium were the dominant genera. Among the 337 isolates, the endophytic fungi of F. sinkiangensis were the most, The Shannon-Wiener biodiversity index (H') associated with roots of F. fukanensis was the highest, reached 1.85. The highest Sorensen coefficient ( Cs) was between leaf of F. sinkiangensis and leaf of F. ovina, reached 0.75. From the result, endophytic fungi were widely distributed in six Ferula, there are some notable differences between distribution and composition of the endophytic fungi isolated from different issues and different species of Ferula, show a certain degree of species and tissue preference. The results obtained in this study will provide realistic basis and theoretical basis for further study the secondary metabolites of endophytic fungi associated with Ferula, and the relationship between endophytic fungi and their host plants.
Biodiversity ; Ecology ; Endophytes ; isolation & purification ; Ferula ; microbiology ; Fungi ; classification ; isolation & purification ; metabolism

Objective To clarify the protective effect of nrsodeoxycholic acid ( UDCA ) on endoplasmic reticulum stress-mediated apoptosis in pancreatic β-cell of streptozotocin ( STZ )-induced diabetic rats.Methods Rats( n =40) received a single injection STZ( 50 mg/kg) intra-peritoneally and formed a β-cell injury model.Weight-matched normal rats( the control group,n =10 ) were injected with the buffer alone.STZ-treated rats with persistent random blood glucose higher than 16.7 mmol/L for 1 week were considered as diabetic status( n=14 ),then divided randomly into STZ-induced diabetes mellitus ( DM ) group ( n =7 ) and UDCA-treated DM group ( n =7 ).UDCA (40 mg· kg- 1,d-1 ) was administered daily by intragastric intubations throughout the experimental period (30 d).During the entire experiment,blood glucose in all rats was assessed.By the end of the experiment,all rats were sacrificed with the pancreas removed and the blood sample collected immediately.Fasting insulin levels were assayed by radioimmunoassay.The morphological changes of pancreatic β-cells apoptosis were determined by TUNEL assay.RNA in pancreas was abstracted and microarray containing 89 pieces of apoptosis related genes was applied.The related gene expressions were detected by RT-PCR and Western blot.Results The concentration of blood glucose in diabetic rats was gradually decreased after UDCA treatment,but at the end of the experiment it was still higher than that in the normal control group.The treatment with UDCA raised the fasting insulin level in diabetic rats,but this concentration was significantly lower as compared to the control group.Based on TUNEL stained tissue sections,the percentage of β-cell apoptosis of UDCA-treated DM group was significantly lower than that of STZ-induced diabetic group(P＜0.05 ).Among 89 genes,42 genes up-regulated and 46 genes down-regulated in diabetic rats,some of which were ameliorated by UDCA treatment.The expressions of Caspase-3,Bax,Bip,and CHOP mRNA in pancreas of DM group were significantly up-regulated as compared with those in the control group ( P ＜ 0.05 ) ; while the expression of Bcl-2 mRNA was markedly down-regulated (P＜0.05 ).However,these parameters in the U DCA-treated animals showed a marked improvement.Conclusion Ursodeoxycholic acid seems to protect pancreatic β-cell from apoptosis in STZ-induced diabetes by attenuating the severity of endoplasmic reticulum stress.

Objective To improve the orotracheal intubation verifying technique and reduce the complication of backflow in rat experiment.Methods A new position evaluation of anti-backflow device was designed and made of safety IV catheter and closed IV catheter system.60 adult male Sprague Dawley rats 216±20 g were randomly assigned to two groups: group A (n=40) for verifying placement, group B (n=20) for anti-backflow test.Group A was further divided into group A1 using self-designed positioning device, group A2 using aerosol, group A3 taking cotton fiber for positioning judgment.The group B was divided into two subgroups, B1 and B2, counting escaped bubbles as a means of positioning observation, the difference is that group B1 using frustum of a cone shape anti-backflow device, while the group B2 using common airway tube.Routine endotracheal intubation was performed to observe and record the time of positioning, the location of exhalation phase, and the length of inspiratory phase countercurrent water column.The group A1 further performed tracheotomy under direct vision clearly to confirm the anatomic positioning status.Results During the exhalation cycle,three or more bubbles were observed to escape continuously, indicating that the intubation tube was properly placed and open in the airway.Positioning time: It took 1.75±1.02 respiratory cycles in group A1,3.30±0.95 respiratory cycles in group A2 and 4.10±0.99 respiratory cycles in group A3 to complete the assessment the positioning status.There was no statistically significant difference between groups A2 and A3 (P> 0.05).The time needed for group A1 was significantly shorter than that of groups A2 and A3 (P < 0.01).The longest countercurrent water column length in group B1 was 3.23±0.53 cm, and 8.48±1.01 cm in the group B2.Conclusions The new designed anti-backflow positioning evaluation device is a simple and convenient appliance to evaluate the location of orotracheal intubation in rat experiment.It can effectively improve the positioning efficiency and has practical application value.

Amblyopia greatly affects the physical and mental development of children. Acupuncture is effective for amblyopia, though its mechanism remains unclear. This article summarized the mechanism of acupuncture treatment of amblyopia from the perspectives of morphology of neurons in visual cortex, visual electrophysiology, and molecular biology, etc. It was found that acupuncture may treat amblyopia through repairing the morphological and ultrastructural damages of neurons in visual cortex, promoting the electrical activities in visual pathway and visual cortical neurons, and modulating the synthesis and expression levels of factors involved in visual system. Nevertheless, further studies are required to unveil the mechanism of acupuncture treatment of amblyopia.