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

Objective To investigate the prognostic factors in acute myeloid leukemia subtype M2 (AML-M2) patients with chromosome translocation of 8 and 21. Methods By using G-banding analyses karyotype and combining the clinical data, prognostic factors in 94 cases of de novo adult AML-M2 in our hospital from 2001 to 2007 were retrospectively analyzed. Results Chromosome 8 and 21 translocation were identified in 53.2 % (50/94) of AML-M2 cases. In the patients with other aberrations in addition to t(8;21), their complete remission (CR) rates, overall survival (OS) was lower than the patients with sole t (8;21) and normal karyotype(P <0.05). And the patients with sole t (8;21) whose CR rates and OS had no difference with patients with normal karyotype (P>0.05). The patients were divided into 3 subgroups (low index, less than 2.5;intermediate index, 2.5-20;high index, 20 or more) according to WBC index. The CR had no difference among the 3 subgroups, but the OS of the 3 subgroups was different (P<0.05). The OS in the lowest index group was longer than that in the others. Conclusion Cytogenetically, 53.2% cases had chromosome 8 and 21 translocation, 46 % cases had t(8;21) with additional chromosomal abnormalities, and the main additional abnormalities were loss of a sex chromosome (LOS). t(8;21) AML-M2 patients with additional chromosome abnormalities had low complete remission rate and shorter survival time, and its prognosis was poorer. WBC index have no influence on complete remission rate but effected the survival time.

ObjectiveTo establish a method for content determination of total iridoid compounds and baldrinal and 11-ethoxyviburtinal from Valerianae Jatamansi Rhizoma et Radix; To determine the contents of total iridoid compounds and baldrinal and 11-ethoxyviburtinal in Valerianae Jatamansi Rhizoma et Radix from three medicinal origins.Methods UV spectrophotometry was applied, 11-ethoxyviburtinal (cyclopentane-pyran-7-formaldehyde, 4-ethoxy methyl) was set as the reference substance, and the content of total iridoid compounds was determined at 288 nm. HPLC method was used to simultaneously determine the contents of baldrinal and 11-ethoxyviburtinal. The HPLC analysis was performed on a Phenomenex Luna C18 column (250 mm×4.6 mm, 5μm). The mobile phase was composed of acetonitrile-water in gradient elution at a flow rate of 0.95 mL/min. The detection wavelength was 288 nm and the column temperature was 30℃.Results The total iridoid compounds, baldrinal and 11-ethoxyviburtinal were in good linearity within the ranges of 2.088–14.616μg/μL, 74.88–224.64μg, and 41.6–249.6μg, respectively. This method was precise, and with good repeatability, stability and recovery rate.Conclusion The method is accurate, simple, rapid, which can be used for the quality control of Valerianae Jatamansi Rhizoma et Radix.

BACKGROUND: Bone exposed wounds are frequently required to deal with in orthopaedic surgeries, involving the treatment of open fractures, bone tumors, osteomyelitis, and many other diseases, in which the defect of soft tissue caused by open fractures is the most difficult to deal with. Conventional debridement or negative pressure closed drainage technology is difficult to make bone exposed wounds heal, and the process is extremely cumbersome, during which,patients suffer a lot of pain.OBJECTIVE: To evaluate the advantages and disadvantages of the various types of dressings, and review the application of new hydrogel dressing in bone exposed wounds based on its advantages, such as keeping wound environment moisture, restoring skin physical barrier, contributing to routine dressing change.METHODS: A computer-based online retrieval of PubMed and CNKI databases was performed to search papers published between 2000 and 2016 using the key words hydrogel dressing, bone exposed wound, traditional wound dressing, antibiotic in English and Chinese, respectively. A total of 55 papers suitable for final analysis from the application of traditional and new dressings in bone tissue engineering were reviewed.RESULTS AND CONCLUSION: The treatment of bone exposed wounds involves the treatment of many diseases, such as open fractures, bone tumors, osteomyelitis, which is still an orthopedic problem to solve. The novel hydrogel dressings with unique advantages are able to provide better plans for bone exposed wounds, and the use of these dressings solves the regeneration and repair of exposed bone, and improves the infection of antibiosis. In addition, the gel dressings currently have become a hot spot of research because of the characteristics of sustained-release.

Objective BRAFV600E mutation, RET/PTC rearrangement, and the concomitant of Hashimoto's thyroiditis(HT) could influence clinicopathological features of papillary thyroid carcinoma (PTC).This study is to investigate the distribution of three factors in PTC and to analyze their associations with clinicopathological characteristics.Methods Fine-needle aspiration samples were collected in a total of 122 conventional PTC patients, who were confirmed by surgery.The clinicopathological features were collected to analyze its association with different factors.BRAFV600E mutation and RET/PTC rearrangement were detected by pyrosequencing and Taqman-qPCR, respectively.Results BRAFV600E mutation was significantly correlated with an older age and a less coexistence with HT(P＜0.05).In contrast, RET/PTC rearrangement was more prevalent in young patients and was associated with the concomitant of HT(P＜0.05).In the age of ≥20 year and＜45 year groups, BRAFV600E mutation was significantly associated with extrathyroidal invasiveness.RET/PTC rearrangement was significantly associated with bilateral lymph node metastasis and the number of metastatic lymph node.Conclusions The distribution of three factors were different in PTC patients.In addition to the age at diagnosis, all of three factors should also be considered together to analyze the association of clinicopathological features of PTC.

Objective To evaluate the effectiveness of mouth care combined with intestinal and endoscopic working channel washing for ERCP related cholangitis.Methods A total of 573 patients diagnosed as having obstructive jaundice were randomized into three groups,190 cases in the control group,192 in the saline group and 191 in the amikacin group.Clinical and laboratory data were collected before ERCP and ERCP related cholangitis were recorded.Results There were no significant differences among the three groups in sex,age,the level of obstruction,the category of obstruction,total bilirubin or WBC counting.The incidences of ERCP related cholangitis were 21.1％ (40/190),13.5％ (26/192) and 4.7％ (9/191)in control group,saline group and amikacin group respectively,which was significantly different (x2 =22.409,P =0.000 ).The incidences of ERCP related cholangitis were 19.5％ ( 65/333 ) and 4.2％(10/240) in patients diagnosed as having hilar duct obstruction and low positioned biliary obstruction respectively (x2 =27.175,P =0.000).There was no significant difference in ERCP related cholangitis between benign and malignant biliary obstruction.Subgroup of hilar duct obstruction showed the incidences of ERCP related cholangitis were 29.7％ (33/111 ),20.5％ (24/117)and 7.6％ (8/105)in the control group,the saline group and the amikacin group,respectively (x2 =16.905,P =0.000).Conclusion The incidence of ERCP related cholangitis is relatively higher in patients with hilar duct obstruction.Mouth care combined with intestinal and endoscopic working channel washing can effectively reduce the incidence of ERCP related cholangitis,especially in the amikacin group.

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.

Objectives To search for the pathogenesis of vaseular endothelial injury and its clinical significance.Methods 36 patients with acute lower respiratory trect infection(ALRI)and 30 healthy controls were envolved. Circulating endothelial cells (CEC)and nitric oxide (NO) levels were tested.Results Circulating nitrite/nitra levels,the stable metabolic products of NO,were found to be significantly higher in the patients with ALRI (44.6?22.6umol/L) than that in the controls (24.5?14.1umoI /L, P

Adult ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cerebral Ventricle Neoplasms ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Fourth Ventricle ; Ganglioglioma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Nerve Tissue Proteins ; metabolism ; Oligodendrocyte Transcription Factor 2 ; Rosette Formation ; Synaptophysin ; metabolism ; Young Adult

Objective To evaluate the clinical outcomes of arthroscopic reconstruction of anterior cruciate ligament (ACL) with femoral tunnel made by three-portal technique.Methods From February 2014 to May 2015,fifty-five ACL rupture patients (38 males and 17 females) with mean age 31.46±7.34 (range,16 to 48) who underwent single-bundle ACL reconstruction with femoral tunnel made by three-portal technique by the same surgeon were followed up.The average preoperative duration was 15.39±18.57 (range,5 to 180) days.All patients presented the positive Lachman test,of which 32 patients had 1 grade pivot-shift test,15 patients 2 grade and 8 patients 3 grade.At final follow-up,the International Knee Documentation Committee (IKDC) and Lysholm scores were used to estimate knee joint function,while the Lachman test and pivot-shift test were used to estimate knee joint stability.Results All patients were followed up for 26.73±5.23 (range,21 to 34) months.The femoral tunnel positions were accurate without fracture of the back walls in all patients.The mean length of femoral tunnel was 38.58±3.76 (range,35 to 42) mm,and the mean angles of femoral tunnel on coronal plane was 49.32°±2.45° (range,45° to 52°).All patients presented negative Lachman test.Fifty-one patients presented negative pivot-shift test,while the remaining 4 patients had 1 grade of the rotational lax-ity.Forty-nine patients showed normal range of motion,while two had 5° flexion deficit,one had less than 5° flexion deficit and three had 5° hyper-extension loss.Furthermore,the IKDC score significantly was improved from 73.31 ±5.83 preoperatively to 94.63±3.16 at the latest follow-up (t=10.14,P＜0.001).The Lysholm score significantly was increased from 66.35±8.91 preoperatively to 95.32±4.25 at final follow up (t=12.76,P＜0.001).Condusion Arthroscopic reconstruction of ACL with femoral tunnel made by three-portal technique is an reliable technique with the advantage of simple procedure and accurate femoral tunnel position in surgery.