OBJECTIVE: To investigate the expression efficiency of exogenous gene mediated by different serotypes of adeno-associated virus (AAV) vectors in retina, and to compare the expression efficiency of AAV vector and two kinds of promoters commonly used in ophthalmology after transfection into mouse retina, so as to provide the basis for selecting appropriate AAV vector and promoter for gene therapy of retinitis pigmentosa. METHODS: AAV2/2, AAV2/5, AAV2/8 and AAV2/9 were prepared. The C57BL/6J mice were injected subretinally with 1 μL purified AAV vectors (1.00×10¹³ mg/L). Then the mice were killed 2 or 4 weeks after treatment, and the eyes were enucleated for frozen section. The expression of green fluorescent protein (GFP) was observed under the confocal microscope. Two kinds of promoters, CMV and CAG, were selectd, and the expression of AAV2/8-GFP-CMV and AAV2/8-GFP-CAG was observed under confocal microscope. RESULTS: No bacterial infection or immune response were seen in the injected mice. 2 weeks after injection, the GFP green fluorescence of AAV2/8 and AAV2/9 in the mouse retina was obvious, which indicated that the GFP green fluorescence of AAV2/8 and AAV2/9 was high after transfection into the mouse retina. In these two serotypes, GFP green fluorescence of AAV2/8 was mainly concentrated in photoreceptor cells while AAV2/8 was expressed in the whole retina, indicating that AAV2/8 was more specific to photoreceptors. Further experiments on AAV2/8 showed that the GFP green fluorescence of the mouse retina was obvious 4 weeks after injection, indicating that the exogenous gene mediated by AAV2/8 could be stably expressed in vivo. For CMV and CAG promoters, CMV promoter was expressed stronger in retinal pigment epithelium (RPE)cells, while CAG promoter was stronger in photorecepters. In photorecepters, CAG promoter was expressed almost the same as CMV promoter, while CMV promoter was stronger in RPE cells. CONCLUSION AAV vectors could express transgene robustly in retinal cells; Among several AAV serotypes, AAV2/2 and AAV2/5 showed weaker GFP fluorescence than AAV2/8 and AAV2/9. AAV2/9 showed expression in each layer of the retina including ganglion cells. AAV2/8 was more specific for photoreceptor; CAG promoters had higher specificity for photoreceptors than CMV promoters.
Animals ; Dependovirus/genetics* ; Genetic Vectors ; Mice ; Mice, Inbred C57BL ; Retina ; Serogroup ; Transduction, Genetic

The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Glucuronidase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Scutellaria baicalensis ; chemistry ; enzymology ; genetics ; Sequence Alignment

Morus alba L. (mulberry) is a well-known deciduous tree, belonging to the genus of Morus of Moraceae famlily. Its leaves, twigs, roots (bark) and fruits are widely used in the traditional Chinese medicine. The active constituents of mulberry contained flavonoids, alkaloids, steroids, coumarins, with the significant hypoglycemic, hypolipidemic, antihypertension, anti-oxidation, anti-inflammatory, anti-bacterial, anti-tumor and immunomodulatory activities. This review summarized the research progress of the major pharmacological activity, pharmacokinetics and drug-drug interaction based on CYPs and transporters of mulberry and its active constituents.
Alkaloids ; pharmacology ; Coumarins ; pharmacology ; Flavonoids ; pharmacology ; Fruit ; chemistry ; Herb-Drug Interactions ; Humans ; Medicine, Chinese Traditional ; Morus ; chemistry ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Steroids ; pharmacology

OBJECTIVE:To provide reference for improving the rational drug use of China. METHODS:By referring to In-dia,South Africa and Argentina's policy in this regard,the rational drug policy rules and supporting measures in the national drug policy of these states were analyzed and summarized to put forward related policies for China. RESULTS & CONCLUSIONS:Pro-moting the rational drug use required multi-angle managements towards the supply-side,the demand-side as well as third-party, and relevant supporting measures needed to be improved. Firstly,the managements on the supply-side is particularly important,in-cluding measures such as strengthening prescription management,formulating and implementing the standard treatment guidelines, intensifing the functions of pharmacists,establishing serious irrational drug use and specific diseases medication management sys-tem;then,the demand-side needs the investigation and feedback about the patients with drug use to eliminate irrational drug use, and through the internet and other media to raise public awareness of the medicine information as well;besides,the third-party needs to establish a non-profit organization to regulate pharmaceutical advertisements,and increase information about rational drug use;finally,improving the rational drug policy also needs to improve needs to promote the separation of medicine,reform Medi-care payment and other supporting measures.

OBJECTIVE:To provide reference for the price management of medicines in Chinese mainland by inquiring into rel-ative management in Hong Kong. METHODS:Based on the steps of price management and the value chain of medicines in Hong Kong,the management mechanism of medicine pricing,circulation and compensation were researched. RESULTS & CONCLU-SIONS:The prices of the medicines in Hong Kong are mainly determined by the tender between Hospital Authority and the pharma-ceutical suppliers. In circulation,medicine payer as main body in bidding and purchase conducthigh qualitybidding andmasspurchase,and price remained the same in public hospital. On the basis of the purchase price,the price that actually paid by pa-tients is the result of government compensation. Chinese mainland can draw lessons from such drug bidding mode in Hong Kong which will lead health care department to act as the main body in bidding,weaken the relationship between the hospitals and the drug dealers or manufacturers,and expand the government's financial investment in public medical institutions to improve reim-bursement.

OBJECTIVE:To explore price management mechanism of medical service in Macau of China,and to provide refer-ence for the formulation of price management mechanism of medical service in the mainland area of China. METHODS:From two key links of pricing and compensation,price management mechanism of medical service in Macau was introduced. RESULTS &CONCLUSIONS:In respect of pricing,Macau Health Bureau as pricing subject set up guide price for public health institutions by marginal cost pricing,which focused on technology value and labor value of medical staff,and graded pricing for health institu-tions at various levels. In respect of compensation,compensation subjects included government,social security funds,charitable or-ganization,etc. The government provided fixed-rate compensation for citizen by allocating funds for public health institutions,so as to improve compensation level for primary health institutions. The social security funds extended sick benefit to insured patients, and compensated patients at fixed rate. Charitable organization subsidized private health institutions to reduce medical service price. Finally,actual price has been formulated through pricing and compensation links,and the experience is worthy of learning by the mainland of China.

Morus alba L. (mulberry) is a well-known deciduous tree, belonging to the genus of Morus of Moraceae famlily. Its leaves, twigs, roots (bark) and fruits are widely used in the traditional Chinese medicine. The active constituents of mulberry contained flavonoids, alkaloids, steroids, coumarins, with the significant hypoglycemic, hypolipidemic, antihypertension, anti-oxidation, anti-inflammatory, anti-bacterial, anti-tumor and immunomodulatory activities. This review summarized the research progress of the major pharmacological activity, pharmacokinetics and drug-drug interaction based on CYPs and transporters of mulberry and its active constituents.

Objective To investigate the changes and clinical significance of platelets aggregation rate of patients with coronary heart disease before and after percutaneous transluminal coronary stenting Methods There are 13 patients underwent coronary angiography and 20 patients underwent percutaneous transluminal coronary stenting Their blood samples were taken from the coronary sinus and coronary artery before and after interventional therapy Results The level of platelets aggregation rate in percutaneous transluminal coronary stenting group was higher after interventional therapy ( P

Objectives To investigate the clinical significance of degree Ⅲ meconium-stained amniotic fluid in term pregnancy.Methods Data of 857 full-term gravidas underwent cesarean section for suspected fetal distress in Peking University First Hospital from October 1,2012 to September 30,2014 were retrospectively analyzed.All patients were divided into three groups according to the diagnosis of suspected fetal distress:meconium-stained amniotic fluid group (Group 1,n=172),fetal heart abnormality group (Group 2,n=623) and meconium-stained amniotic fluid in combination with fetal heart abnormality group (Group 3,n=62).General information and fetal prognosis of the three groups were analyzed.Factors that might affect the prognosis of newborns were analyzed between the infants with or without acidosis,asphyxia or aspiration pneumonia.Analysis of variance,independent-samples t test,Chi-square test and rank-sum test were used as statistical methods.Results There were no significant differences in maternal age,gestational age at delivery,parity,nuchal cord loop(s),pH value of umbilical arterial blood and gender of newborns among the three groups (all P＞0.05).The neonatal Apgar score,neonatal asphyxia rate and rate of admitting into pediatric ward within 24 hours after birth in Group 1 were similar to those of Group 2 and 3 [1 min Apgar score:9.9±0.7 vs 9.8±0.7 and 9.8±0.7,F=0.322;5 min Apgar score:10.0±0.3 vs 10.0±0.2 and 10.0±0.0,F=0.517;neonatal asphyxia rate:1.7％ (3/172) vs 2.1％ (13/623) and 1.6％ (1/62),x2=0.129;rate of admitting into pediatric ward:16.3％ (28/172) vs 14.3％ (89/623) and 11.3％ (7/62),x2-0.978] (all P＞0.05).The incidence of neonatal aspiration pneumonia in Group 1 was higher than that of Group 2 and 3 [4.7％ (8/172) vs 3.2％ (2/62) and 1.0％ (6/623)],but the differences had no statistical significance (F=10.680,P=0.050).Conclusions Both degree Ⅲ meconium-stained amniotic fluid and abnormal fetal heart rate have the same indicating significance and should be treated actively to lower the risk of poor prognosis of the newborns.

Aim To observe the protection effect of Tribulus terrestris saponin monomer B (TTSMB) on cadiocytes impaired by hypoxia-reoxygenation (H/R).Methods Cadiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, H/R group,GSTT 100 mg·L~(-1) group and TTSMB 10,1,0.1 nmol·L~(-1) group.Morphocytology change of cadiocytes was observed after the treatment.Cadiocyte survival rate was detected with MTT colorimetric method.Levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), malondialdehyde (MDA), and activity of superoxide dismutase (SOD) were determined. Apoptosis rate was detected with flow cytometry.Expression of caspase-3 was examined with western blot.Results Compared with the control group, the survival cell number in H/R group decreased obviously, content of CK,LDH, AST, MAD increased, and activity of SOD decreased (P<0.01).Compared with the model group, the survival cell population increased in TTSMB 10,1,0.1 nmol·L~(-1) group (P<0.05 and P<0.01).Contents of CK, LDH, AST, MAD decreased, whereas the activity of SOD increased (P<0.05 and P<0.01).Apoptosis rate and expression of caspase-3 also reduced in TTSMB 10,1,0.1 nmol·L~(-1) group.Conclusion TTSMB has significant cadiocytes protective effect against H/R and inhibits cadiocyte apoptosis,and the mechanism depends on the effect against oxygen free radical.