Objective To clone, express and purify human PIF1 protein and analyze its ATPase activity. Methods The PIF1 cDNA was amplified by PCR from HeLa cell cDNA library and inserted to pET24b with histidine tag at its terminus to form pET24b-PIF1 plasmid. The recombinant pET24b-PIF1 plasmid was transformed to RosettaTM 2 (DE3) and the expression of PIF1 protein was monitored by SDS-PAGE analysis. By using fast protein liquid chromatograph (FPLC) system, the PIF1 protein was purified by affinity chromatograph and gel filtration. The ATPase activity of PIF1 was checked by thin layer chromatograph(TLC). Results The PIF1 protein was successfully cloned and expressed in E.coli. Conclusions The purification procedure of PIF1 protein was established using FPLC. The overexpressed and the purified PIF1 helicase has DNA and Mg2+ dependent ATPase activity.

Treating different diseases with the same method is a unique original thinking in Chinese medicine, which embodies the spirit of treatment based on syndrome differentiation. We briefly reviewed the origin and development of this concept. We also reviewed that thinking of combination of disease and syndrome is its premise and foundation. We put forward the conditionality of diseases in treating different diseases with the same method by cutting-in modern biological basic researches, that is to say, one kind specific diseases or disease of one specific system. We emphasized the importance of diseases conditionality in treating different diseases with the same method, which was of great significance in studies on combination of disease and syndrome, correspondence of prescription and syndrome, and modern biological basic researches of treating different diseases with the same method.
Humans ; Medicine, Chinese Traditional ; Prescriptions ; Research

Acute Disease ; Adult ; Angioplasty, Balloon, Coronary ; adverse effects ; Humans ; Hypopituitarism ; etiology ; therapy ; Male

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.
Caffeic Acids ; analysis ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Hydroxybenzoates ; analysis ; Lonicera ; chemistry ; Quinic Acid ; analogs & derivatives ; analysis

ObjectiveTo evaluate the reliahility and validity of the chronic HBV-infections related stigma scale.MethodsThe initial items and construct of the scale were developed according to theoretical analysis and interviews of experts and patients.A total of 151 patients with chronic HBV-infection were administered by convenient sampling method in this pilot study. The reliability and the validity of the scale were then evaluated.ResultsThe response rate of the scale was 94.5％.The Cronbach α coefficients of all dimeusions ranged from 0.75-0.87.The results of correlation analysis showed that there were higher correlation coefficients ( r ranged from 0.62-0.86) between items and their hypothesized subscales than those with other subscales ( r ranged from 0.14-0.55).The scale distinguished between patients with low subscale scores ( the subscale scores were ( 1.89 ±0.30 ),( 1.86 ± 0.29 ),( 1.96 ± 0.23 ),( 2.29 ± 0.45 ),( 1.59 ± 0.42 ) independently) and those with high subscale scores(the subscalc scores were (3.62 ±0.44),(3.99 ±0.41 ),(3.79 ±0.37),(4.13 ±0.34),(3.10 ±0.53 ) independently) (P ＜ 0.01 ).Confirmatory factor analysis showed that the main indices of goodness of fit CFI was 0.94,NNFI 0.92,RMSEA 0.087.ConclusionThe chronic HBV-infections related stigma has good psychometric properties regarding to reliability and validity.

Objective To compare the effects of antenatal ambroxol and 3-day dexamethasone and 1-day dexamethasone on rats′ fetal lung morphogenesis.Methods Twelve pregnant rats were divided into 4 groups randomly:3-day ambroxol group,3-day dexamethasone group,1-day dexamethasone group and control group,every group had 3 rats.On gestational day 19,cesareans were carried out and the histologic structures of 6 fetal rats lungs of each pregnant rats were observed with light microscope,electronic microscope and image analysis.Results 1.Under the light microscope,compared with control group,fetal rats lung in three treatment groups had more alveolar numbers,larger alveolar space,and thinner alveolar septum(Pa

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.

Objective　To study the probability of using hepatitis D virus (HDV) ribozyme as a kind of anti-hepatitis-C-virus (HCV) gene thera-py drugs. Methods　The natural HDV genomic ribozyme′s stem Ⅳ was optimized and its substrate-binding region reconstructed, thus three recombinant HCV-specific HDV genomic ribozymes RzC1, RzC2 and RzC3 were obtained. HCV RNA 5'-noncoding region and 5'-fragment of C region (HCV RNA5'-NCR-C) were transcribed from plasmid pHCV-neo by T7 phage RNA polymerase in vitro, and radiolabelled at its 5'-end. The trans-cleaving reaction was performed by mixing the ribozymes and substrate at mol ratio 100∶1 under conditions as follows: 37℃, pH7.5, Mg2+ 20 mmol/L and deionized formamide 2.5 mol/L. Percentage of trans-cleaved products were calculated at different time points and used as the activity indicator of the three ribozymes. Results　RzC1, RzC2 trans-cleaved more substrate when the time extended, and got to 24.9%,20.3% after reac-ting for 90 minutes respectively; RzC3 was not able to trans-cleave its substrate. Conclusion　Recombinant HDV genomic ribozymes have the ability to trans-cleave HCV RNA, but the appropriate target sequence should be selected.

OBJECTIVETo explore the clinical outcomes of open reduction and internal fixation with calcaneal locking plates in treating Sanders type II and III calcaneal fractures.

METHODSFrom January 2010 and October 2012, 38 calcaneal fractures with Sanders type II or III were treated with open reduction and internal fixation with calcaneal locking plate. According to the Sanders classification, 15 fractures were classified as type II, 23 fractures as type III. The patients were divided into two groups (group A and B) according to the different fixed methods. Sustentaculum tali was fixed with one screw in group A, including 13 males and 5 females, with a mean age of (38.56±8.03) years old (ranged, 25 to 55). And sustentaculum tali was not fixed in group B, including 16 males and 4 females, with a mean age of (42.35±8.29) years old (ranged, 29 to 53). Clinical effects were evaluated according to the changes of Böhler's angle and the Maryland Foot Score and VAS score.

RESULTSAll patients were followed up from 12 to 20 months with a mean of 14 months. Böhler's angles and subtalar joints obtained satisfactory reconstruction in all patients. One year after operation, the mean Maryland Foot Score was 88.61±7.59 in group A; and was 82.40±9.24 in group B; Maryland Foot Score of group A was higher and foot functional rehabilitation was better than group B. The mean VAS score was 13.39±11.47 in group A; and was 22.50±13.10 in group B; VAS score of group A was lower and foot pain was less than group B.

CONCLUSIONSustentaculum tall screw fixation has advantages of strong fixed strength, high stability, less postoperative pain, rapid functional recovery in treating Sanders type II and III calcaneal fractures.

Adult ; Bone Plates ; Bone Screws ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function

Objective To investigate the influences of gender,age,and body mass index (BMI)on the arterial stiffness in hypertensive patients.Methods A total of 1026 hypertensive patients were enrolled in this study from 6 community health centers in Baoshan District and Shanghai Third People's Hospital.The Related medical histories and carotid-femoral(C-F) pulse wave velocity (PWV) were recorded and PWV were re-tested in 672 patients after 1 year's follow-up.The C-F PWV was analyzed from different group of Gender,age and BMI at baseline and after follow up.Results (1)C-F PWV of the male hypertensive patients were higher than that of female's and the corresponding figures were (13.63 ±3.07)m/s and (12.59±2.85)m/s(P ＜0.01).After one year's follow up,C-F PWV was increased in both male and female with corresponding figures of (14.11 ± 3.58) m/s and (13.51 ± 3.56) m/s (P ＜ 0.05).(2) The C-F PWV was increased with age.The corresponding figures were [(11.11 ± 2.00) m/s for age group 30 ～ 50 years old,(12.73 ± 2.77) m/s for age group ＞ 50 ～ 70 years old,and (14.74 ± 3.29) m/s for age group ＞ 70 years old,all P ＜0.01].After one year's follow-up,C-F PWV of three age groups was increased and the corresponding figures were [(11.36±1.78)m/s(P ＞0.05),(13.39±3.33)m/s(P ＜0.01),and (15.68 ±3.94)m/s(P ＜0.05)].The increased range of C-F PWV (from high to low) was age group ＞ 70,age group ＞ 50 ～70 and age group 30 ～50.(3) C-F PWV of patients with BMI ≥ 24 kg/m2 was significantly lower than that of patients with BMI ＜24 kg/m2 and the corresponding figures were [(12.93 ±2.89)m/s and (13.39 ±3.18)m/s,P ＜0.05].After one year's follow up,C-F PWV was increased and the figures were [(13.54 ± 3.54) rn/s and (14.27±3.61) m/s,respectively(P ＜0.01)].Conclusions Except BMI,gender and age were the important influence factors on arterial stiffness.