OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

?AlM: To evaluate the long - term effects of laser peripheral iridectomy ( LPl ) and trabeculectomy in treating early chronic angle-closure glaucoma.
?METHODS: Ninety-eight patients (102 eyes) with early chronic primary angle-closure glaucoma were randomly divided into two groups. Group A of 50 patients (54 eyes) was treated with LPl and group B of 48 patients (48 eyes) with trabeculectomy. After 3 - 8y of follow - up observation, comparison would be made from the perspectives of postoperative eyesight, intraocular pressure, anterior chamber angle, visual field and cup/disc ratio ( C/D) .
?RESULTS:ln group A, 24 eyes with eyesight declining, 22 eyes with theintraocular pressure>21mmHg (1mmHg=0. 133kPa), 21 eyes with chamber angle synechia >180o, 21 eyes with visual field narrowed, 21 eyes with C/D ratio enlarged. The results of group B for the same items were 10, 5, 4, 4, 4 eyes respectively. The comparative difference was statistically significant (P<0. 05).
?CONCLUSlON:Good effects will be achieved for early-stage chronic angle - closure glaucoma with surgical method. Trabeculectomy is obviously better than LPl for the long-term effects.

Tumor vaccine is one of the most promising therapeutic strategies in tumor immunotherapy. It promotes the antigen presentation process by delivering tumor antigen and then activates the anti-tumor immune response. As a new class of vaccines, messenger RNA (mRNA) vaccines can activate the immune system to achieve the purpose of immunotherapy by delivering the mRNA sequence of a specific antigen into the body and expressing the corresponding antigen protein. Compared with traditional vaccines, mRNA vaccines have the advantages of a short production cycle, high effectiveness, and strong immunogenicity. In recent years, the application of mRNA vaccines in tumor immunotherapy has attracted widespread attention, but the instability and low delivery efficiency of mRNA limit its application. Nano delivery system can effectively solve the problem of mRNA vaccine delivery, greatly promote the research process and clinical application of mRNA tumor vaccines, and has become a hot spot in the research of mRNA vaccines. In this review, we introduced the mRNA tumor vaccines, focusing on the application of nano delivery system in mRNA tumor vaccines, in order to provide new ideas and new methods for the efficient delivery of mRNA tumor vaccines and tumor immunotherapy.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Diseases ; diagnostic imaging ; Tomography, Spiral Computed ; methods

AIM To study the therapeutic effects of Deketoprofen trometamol (D KPT) on adjuvant arthritis (AA) and its side effects on gastroduodena. METHEDS The change of primary and secondary inflammatory and the injury effect of gastroduodena treated with D KPT were observed. The effect of D KPT on prostaglandin E 2 (PGE 2) produced by peritoneal macrophage (PM) on AA rats in vitro was also observed. RESULTS D KPT (2 5, 5, 10 mg?kg -1 ) significantly inhibited primary inflammatory, secondary inflammatory and multiple arthritis of AA rats. D KPT(10 -8 ,10 -7 ,10 -6 ,10 -5 ,10 -4 mol?L -1 )inhibited the elevated PGE 2 released from PM of AA rats in vitro . D KPT (10 mg?kg -1 ) significantly injured gastroduodena on AA rats, but the degree of injuries was less than that of ketoprofen (KP) (10 mg?kg -1 ). CONCLUSION D KPT has therapeutic effects on AA rats, and its injury effect on gastroduodena is less than that of KP.

Objective To discuss the diagnostic value of dynamic contrast-enhanced MRI in differentiating vertebral metastatic tumors from benign deseases after studying the early changes of signal intensity.Methods Twenty patients were scanned by dynamic contrast-enhanced MRI.The parameters of signal intensity were measured and calculated,and time-intensity cures(TICs)were drawn.Then,they were compared with those of normal verte- braes,the characteristics were analyzed.Results The parameters of vertebral metastatic tumors:peak signal intensi- ty,signal intensity minimum signal intensity of post-peak,peak slope,initial ratio of enhancement,maximum ratio of enhancement was respectively(50.61?11.38),(46.75?10.23),(0.73?0.12),(0.33?0.08),(1.03?0.31); that of normal vertebraes was respectively(40.53?12.12),(34.72?14.06),(0.31?0.11),(-0.25?0.05), (0.69?0.28);and ten normal cases showed negative in initial percent of enhancement,but only two abnormal ver- tebraes showed that.All of the above had significant statistical difference.Four types of TICs were concluded:early rapid rise and early rapid descent followed by steady phase(type A),persistently rise(type B),rise phase followed by steady phase(type C),rise phase followed by rapid descent phase(type D).TICs of abnormal vertebraes were respec- tively 6,5,5,5 cases;types of normal vertebraes were mainly type C,15 cases.Type A and type B and the type with the characteristic of early rapid descent phase were features of abnormal vertebraes;type C was the feature of normal ones.Conclusions Dynamic contrast-enhanced MRI could quantitatively show the characteristics of early changes of signal intensity of vertebral metastatic tumors,which were significant statistically different from those of normal ver- tebraes.Dynamic contrast-enhanced MRI was worth performing in the early diagnosis of vertebral metastatic tu- mors.

Clinical data of 50 patients with gastric injury after blunt abdominal trauma(gastric rupture in 20 and gastric laceration in 30)were retrospectively analyzed.All the patients with gastric rupture had a satiate stomach.which was not found in the patient with gastric laceration.Forty-nine patients experienced combinant injury.The most common combinant injury of gastric laceration was liver and pancreatic injury(15/30 and 10/30),and the most frequent combinant injury of gastric rupture was spleen injury(11/20).Forty-eight patients recovered while 2 died after the treatment with gastric prosthesis.The mortality rate was 4%.Surgical operation,or prosthesis,could be the main treahnent of choice.The management of combinant injuries and complications should be imprta.

A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

To study the effects of major components of Maijunan tablets, puerarin (Pue) and rhynchophylline (Rhy) on the transport of hydrochlorothiazide (Hct) Caco-2 cell monolayer model, the transport parameters of Hct, such as apparent permeability coefficient (P(app) (B --> A) and P(app) (A --> B)) and the ratio of P(app) (B --> A) versus P(app) (A --> B), were studied and compared when Hct was used solely and co-used with Pue and/or Rhy. The effects of drug concentrations, conveying times, P-glyprotein (P-gp) inhibitor verapamil and conveying Liq pH values on the transport of Hct in the above conditions were also investigated. The results indicated that the absorption of Hct in Caco-2 cell monolayer model could be a carrier-mediated active transport, along with the excretion action mediated by P-gp. Pue can decrease the excretion action of Hct mediated by P-gp, and Rhy had no significant effect on the transport of Hct. The co-use of Hct, Pue and Rhy enhanced the absorption of Hct. Meanwhile, conveying Liq pH value had significant influence on the transport of Hct. The absorption of Hct at pH 6.0 was higher than that at pH 7.4.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Absorption ; drug effects ; Biological Transport, Active ; drug effects ; Caco-2 Cells ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Humans ; Hydrochlorothiazide ; pharmacokinetics ; Hydrogen-Ion Concentration ; Indole Alkaloids ; administration & dosage ; isolation & purification ; pharmacology ; Isoflavones ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Time Factors ; Vasodilator Agents ; administration & dosage ; isolation & purification ; pharmacology ; Verapamil ; pharmacology

Objective To investigate the disease situation of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008 so as to explore the relationship between water-related endemic fluorosis and incidence of carotid atherosclerosis disease. Methods A total of 266 participants over the age of 40 from four villages in Zhaozhou county with water fluoride ≥ 1.0 mg/L in a fluorosis area and 283 residents over the age of 40 from four villages in Tailai county with water fluoride ＜ 1.0 mg/L in a nonfluorosis area were investigated. A portable-type B mode color ultrasound was used to examine the left carotid artery of all participants.The carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of carotid atherosclerosis in the 4 fluorosis villages(Xinfeng, Taipingshan, Baochan and Houzheng villages) were 47.3% (35/74), 63.5%(40/63), 73.3%(33/45) and 60.7%(51/84), respectively, and in the 4 nonfluorosis villages(Hala,Qianxing, Sanjia and Ailin villages) were 32.7% (17/52), 32.9% (24/73), 39.2% (31/79) and 30.4% (24/79),respectively. The prevalence rates of carotid atherosclerosis in every villages standardized by age were 47.3% ,63.5%,73.3% ,60.7% and 34.7% ,36.3% ,43.0% ,41.3%, respectively. Statistic method used was Wilcoxon two sample test and the differences were significant(T = 26, P ＜ 0.05 ). Total carotid atherosclerosis positive rate standardized by age in the 4 fluorosis villages was significantly higher than that in the 4 nonfluorosis villages[57.5%(153/266) ,37.8% (107/283), x2 = 21.36, P ＜ 0.01 ]. After standardized by age, the severity of carotid atherosclerosis was significantly different between fluorosis villages and nonfluorosis villages (x2 = 36.15, P ＜ 0.01 ). Conclusion The prevalence rate of carotid atherosclerosis in endemic fluorosis area is higher than that in nonfluorosis area.