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

To discuss the influence of dexamethasion on lL-1β and TNF - α expression in suture - induced rabbit corneal neovascularization ( CNV ) and analyze the potential mechanism.
●METHODS: For 43 healthy rabbits, 40 were randomly selected for establishing CNV model in corneal stroma. The right eyes (group A) were received no medicine and the left eyes ( group B) were injected dexamethasone after successfully establishing the model. The no modeling 3 rabbits were normal control group. The morphologic change of corneal was observed with slit lamp microscope and the areas of CNV was calculated every day, then 8 rabbits were randomly chosen for sacrificing at 1, 4, 7, 14, 21d respectively. The pathological characteristics of CNV were observed after HE staining, and lL - 1β and TNF - α expression was detected by immunohistochemistry.
●RESULTS: CNV was grown at the 4d after suture, and the 7-14d was vigorous growth period. inflammatory cell infiltration appeared after HE staining, and CNV was located at the superficial stroma of cornea. lmmunohistochemistry results showed that lL - 1β and TNF - α expression was gradually increased with prolonged suture time. Compared with corneal stitch group, the rabbits cured by dexamethasone were found with less inflammatory cells infiltrating and neovescularization, moreover, the expression of lL - 1βand TNF-α decreased. There were statistical significance between the two groups (P<0. 05).
● CONCLUSlON: Dexamethasone can inhibit the CNV growth by controlling the inflammation of corneal and restraining lL-1β and TNF-α expression.

With the global development of medical information standard, construction work of traditional Chinese medicine (TCM) information standard system has been promoted rapidly. The framework of ancient Chinese medical literature clinical terminology classification standardization is one of the foundations of TCM language system. Its research will further promote and perfect TCM information standard system. We have adhered to the connection among ancient TCM classification framework, modern TCM and western medicine classification framework. Exploration on framework construction of ancient Chinese medical literature clinical terminology classification standardization was based on previous work of Chinese medicine clinical terminology classification and code standard, reference frame structure of SNOMED CT, research findings of classification standard framework of Chinese medicine clinical terms, andQian Jin Fang.

BACKGROUND:Poststroke depression is one of the most common psychological behavior disorders after stroke and its mechanism remains unclear. Studies have suggested that microRNAs (miRNAs) involved in neurogenesis and synaptogenesis may play an important role in psychology diseases. OBJECTIVE:To observe the expression of miR-137 in the blood and brain of poststroke depression rats and its effect on the behaviors of rats. METHODS:Thirty-six rats were equal y divided into six groups:control, model, agomir-137, agomir-NC, agomir-137+Grin2A and agomir-137+vector groups. Control group had no treatment. Poststroke depression models were established by ligation of middle cerebral artery and chronic mild stimulation in the latter four groups fol owed by receiving an injection of nothing, agomir-137, agomir-NC, LV-CMV-Grin2A or control plasmids into the left lateral ventricle, respectively. RESULTS AND CONCLUSION:We found significantly lower miR-137 levels in the brain and peripheral blood of post-stroke depression rats compared with normal rats. Vertical scores and horizontal scores on the behavior test were significantly higher in the agomir-137 group than the agomir-NC and model groups at 3 weeks after cerebral ischemia;while, sucrose consumption percentage was also higher in the agomir-137 group at the end of 2 weeks after cerebral ischemia. Luciferase assays showed miR-137 bound to the 3’ UTR of Grin2A, regulating Grin2A expression in a neuronal cel line. Grin2A gene overexpression in the brain of post-stroke depression rats noticeably suppressed the inhibitory effect of miR-137 on post-stroke depression. Overal , these findings show that miR-137 suppresses Grin2A protein expression through binding to Grin2A mRNA, thereby exerting an inhibitory effect on post-stroke depression and offering a new therapeutic target for poststroke depression.

AlM: To describe the expression of hypoxia-inducible factor-1α ( HlF-1α) and erythropoietin ( EPO ) in rats' corneal and evaluate its potential effect on corneal neovascularization ( CNV) growth.
METHODS:The young SD rats (3mo) was chosen and randomly divided into 2 groups, which were experimental group and normal control group. CNV model was established by alkali burn, and the length and area of CNV was observed everyday after operation by slit lamp. After that, the expression of HlF-1α and EPO was measured by SABC and RT-PCR methods at 1, 3, 5, 7, and 14d after alkali burn. The data was analyzed by SPSS 20. 0.
RESULTS:The area of CNV was increasing at 1, 3, 5, 7, and 14d after alkali burn, and the peak point appear at 7d. The growth speed was decreased after 14d. SABC method told us that no HlF-1αand very tiny amount EPO was detected at normal rats' corneal. The expression of the two factors increased at 1d after alkali burn in corneal epithelium and endoderm. The results of RT - PCR showed that a few amounts of HlF-1α and EPO mRNA were detected at normal group. The expression of the two factors was increased at 3d after alkali burn, and the peak value was found at 7d, however, it was decreased at 14d. Statistical difference was found at different time (P<0. 05).
CONCLUSlON: HlF- 1α and EPO is closely related to CNV.

Objective: To explore the role of cytosine-phosphate-guanine oligodeoxynucleotide(CpG ODN)in enhan- cing the radiosensitivity to X-ray in mouse with Lewis lung cancer.Methods: The tumor-bearing mouse model was in- duced by injevting Lewis lung cancer cells into the right infra-axillary dermis.Thirty-two C57BL/6J mice were evenly ran- domized into 4 groups.Group A: the control group;Group B: the X-Ray radiation group;Group C: the CpG group; Group D: the CpG plus X-Ray radiation group.Group B was treated with X-Ray radiation only(3 Gy/F,on day 1,3,5, 8,10,and 12;the total dose was 18 Gy);group C was administered with CpG ODN 0.05 mg on day 1,3,5,8,10, and 12;group D was administered with CpG ODN 6h before X-ray radiation.The tumor growth and tumor growth delay (TGD)were observed in all groups.Meanwhile,the pathological change of the tumor tissue was observed with H-E staining method and the apoptosis of tumor cells were examined with the method of TUNEL.Results: The Lewis hmg cancer-bearing model was successfolly established in mice.The tumor volumes of the treatment groups were smaller than that in lhe control group(P

Objective To evaluate and report the performance of PL-11 platelet analyzer.Methods Intravenous blood samples anticoagulated with EDTA-K2 and sodium citrate were tested by the PL-11 platelet analyzer to evaluate the intra-assay and inter-assay coefficient of variation (CV),carry-over rate,accuracy,linearity of the PL-11 platelet analyzer.Platelet aggregation rate of sodium citrate-anticoagulated fasting venous blood collected from 30 physical examinees in outpatient department of The First Affiliated Hospital of Soochow University between February and July,2012 was detected by the PL-11 platelet analyzer and MPG-3E multifunctional double channel blood coagulation analyzer,respectively.The correlation was detected between the PL-11 platelet analyzer and MPG-3E multifunctional double channel blood coagulation analyzer.Results All the parameters were conformed to the standard of Clinical Laboratory Improvement Amendment 88.Both of the intra-assay and inter-assay CV values were less than 5％ ; carry-over rate was less than 1％ ; the accuracy and the linearity was excellent correlated to the result of photoelectric turbidimetry (R2 =0.9439).Conclusions PL-11 platelet analyzer can directly use whole blood dynamics to analyze platelet aggregation process and quantitatively analyze the various components of blood cells (including platelets and red blood cells),the result reports are accurate and reliable.

Objective To summarize the clinical phenotype profiles and mitochondrial DNA mutation in maternally inherited diabetes and deafness ( MIDD ) , and to improve the diagnosis and treatment of this disease in clinical practice. Methods Sixteen patients with MIDD in six families from Peking Union Medical College from 2007 to Dec 2014 were confirmed as carrying the mitochondrial ( mt) DNA 3243 A to G mutation. Sanger sequencing was used to detect the mt DNA 3243 A to G mutation. The peak height G/A ratio was used to determine mutation heteroplasmy levels. Results The patients with early onset of diabetes (35. 0 ± 14. 6 years), deafness, normal or lower body mass index ( BMI) , and maternal hereditary tendency suggested the diagnosis of MIDD. The peak height G/A ratio was significantly different according to the onset age of MIDD [≤25 years (61. 6 ± 20. 17)%;25-45 years (16.59±8.64)%;>45 years(6.37±0.59)%;P<0.01]. The peak height G/A ratio was negatively correlated with the onset age of MIDD(r=-0. 785,P=0. 001). Conclusion Early onset of diabetes with deafness, normal/lower BMI, and maternal hereditary tendency strongly suggests the diagnosis of MIDD. The peak height G/A ratio might provide a simple prediction regarding the onset age and severity of MIDD.

Background Placement of an orbital implant is a main way to prevent orbital atrophy with aging.But its mechanism is under clear.Researchs showed that bone growth factors play important role during the development and repair of bone,especially transforming growth factor-β1(TGF-β1).Objective Present study was to investigate the expression of TGF-β1 protein in orbital bone after enucleation or enucleation with placement of an orbital implant and its function in the mechanisms of preventing and treating the orbital malformed development after enucleation with placement of an orbital implant.Methods Twenty-one age- and weight-matched New Zealand white young rabbits were randomly divided into the enucleation,implant and control groups,and each group including seven rabbits.Eyeball nucleation surgery was performed in the left eyes of 7 1-month-old rabbits,and a spherical orbital implant was inserted after enucleation of the left in matched rabbits in implant group.The left eye of normal rabbits served as controls.The rabbits were sacrificed in 1 month after surgery.The expression of TGF-β1 protein in the left orbital bone was detected using enzyme immunoassay and FITC labelling immunoassay technique in the sections of zygomatic bones.The content of TGF-β1 protein in the left orbital bone tissue was measured by ELISA method.This use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The height and width of orbital in enucleation group were significantly lower than those of implant and normal control groups(height:P=0.00,P=0.00;width:P =0.00,P=0.00).The positive bone cells of both enzyme immunoassay and FITC staining were increased in the implant and control groups in comparison with enucleation group,but the positive response intensity for TGF-β1 was resembled between implant group and control group.ELISA result revealed that the content of TGF-β1 protein in bone tissue was significantly lower in the enucleation group than in implant and control groups(P=0.00,P=0.00).The expression and content of TGF-β1 protein in bone tissue is similar between the implant group and the control group(P=0.41). Conclusion The experiment results indicate that TGF-β1 protein participate in the orbital development.TGF-β1 played important role in the prevention and treatment of enucleation-induced orbital malformation in the eye with placement of an orbital implant.

OBJECTIVETo study the anti-mutation action of acupuncture and moxibustion.

METHODSMice were randomly divided into 6 groups, group 1 (normal control group), group 2 (positive control group), group 3 (prevention group I), group 4 (prevention group II ), group 5 (treatment group I) and group 6 (treatment group II). The mice in the group 2-6 were treated by cyclophosphamide (ip, 50 mg/kg body weight), and in the 3'-6 groups were given acupuncture at "Zusanli" (ST 36) and moxibustion at "Guanyuan" (CV 4). At the end of experiment, all the mice were decapitated and chromosome aberration rate and sister chromatid exchange rate of bone marrow cells were investigated.

RESULTSThe chromosome aberration rate and the sister chromatid exchange rate of bone marrow cells in the positive control group increased significantly as compared with the normal control group, while they decreased significantly in the group 3, 4, 5, 6 as compared with the positive control group (P < 0.01).

CONCLUSIONAcupuncture and moxibustion have anti-mutation action, inhibiting the increase of chromosome aberrations and sister chromatid exchange of bone marrow cells in mice induced by cyclophosphamide.

Acupuncture Therapy ; Animals ; Cyclophosphamide ; Mice ; Moxibustion