Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.

Objective DNA replication increases during tumor cells limitlessly proliferate. To develop a method that can early diagnose and routine screen out bladder cancer, but it does not impair patients by detecting cdc6 in exfoliate cell in urine of patients with bladder cancer. Methods 200 ml fresh urine of patients with bladder cancer or non bladder cancer who voided urine after morning was collected and centrifugated for urine sediment, then total RNA was extracted with TRIZOL reagent from it and detected the expression of cdc6 gene by RT PCR. Results 28 samples were positive and 2 samples were negative in 30samples of bladder cancer, positive rate was 93.3%. Positive rate was 16.7% in 30 samples of non bladder cancer. Compared with the sensitivity of urine cytology, that of cdc6 RT PCR increases markedly ( P

During growth and progression, the microenvironment of tumors suffers a series of abnormal characteristics, which include hypoxia, acid pH, increased oxidative stress, excess glutathione (GSH), as well as certain overexpressed enzymes. Although affect or limit the cancer therapeutic outcomes, these factors provide possible approaches to strategies for cancer detection and novel therapy at the same time. Recently, based on these properties of the tumor microenvironment (TME), various kinds of responsive nano-platforms have been continuously developed and applied in cancer theranostics preliminarily. Thus, this review would introduce the typical features of TME firstly, then detailly summarize the design principles and research progress of corresponding hypoxia-responsive, pH-responsive, redox-responsive, enzyme-responsive, dual-responsive and multi-responsive nano-platforms. Finally, the challenges and the perspectives of the TME-responsive nano-platforms are briefly discussed.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

In lieu of large samples of cases and/or controls with hundreds of markers spreading throughout the human genome, researchers started to notice the dramatic increase of genome-wide association study (GWAS) for complex disorders, in the last 5 years. This paper highlights the statistical challenges in such huge-scale genetic studies, and introduces the analytical strategies and steps for handling GWAS data. Such issues as quality control of data, population stratification, methods available to data analysis and results presentation, replication, as well as the limitations of GWAS studies and the challenges presenting for statistics, are addressed.

Objective To investigate the differences between indocyanine green (ICG) fluorescence imaging plus methylene blue and plus Carbon Nanoparticles Suspension Injection for sentinel lymph node biopsy (SLNB)in breast cancer patients. Methods A total of 134 cases of early breast cancer patients performed SLNB from November 2013 to November 2016 were involved,of which 48 cases were performed with ICG fluorescence imaging plus methylene blue,and another 86 cases plus Carbon Nanoparticles Suspension Injection. Results There was no significant difference between ICG plus Methylene Blue group and ICG plus nano carbon group in terms of detection rate(P>0.05),detected numbers(P>0.05),sensitivity(P>0.05),accuracy(P>0.05)and false negative rate(P > 0.05). Age,and body mass index(BMI)exerted no influence on the detection rate and accuracy of SLNB in two groups(P>0.05). Conclusion ICG Fluorescence Imaging plus Methylene Blue showed similar detection rate , detected numbers , sensitivity , accuracy and false negative rate as it plus Carbon Nanoparticles Suspension Injection for SLNB in breast cancer patients ,and both of them can be performed easily and conveniently.

Adult ; Air Pollutants ; adverse effects ; Humans ; Male ; Military Medicine ; Monitoring, Physiologic ; Noise, Occupational ; adverse effects

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

AIMTo study the effects of indomethacin on interleukin-6 (IL-6) expression stimulated with lipopolysaccharide (LPS) in rheumatoid arthritic patients' synoviocyte.

METHODSFibroblast-like cells (FLS) from rheumatoid arthritic patients' joint tissue were cultured for 24 h and incubated 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated by LPS (1 mg.L-1). After indomethacin or dexamethasone added into the supernatant of U937 cells, FLS was incubated with the super natant for 24 h. The expression of IL-6 protein was detected by radioimmunoassay. The mRNA expression of IL-6 was accessed by RT-PCR.

RESULTSLPS did not obviously affect the growth of FLS, and the protein secretion and mRNA expression of IL-6 were not changed in FLS treated with LPS. The IL-6 secretion and IL-6 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Indomethacin at concentrations of 1 x 10(-7)-1 x 10(-5) mol.L-1 obviously inhibited the protein secretion and mRNA expression of IL-6 in FLS cultured with the supernatant from U937 cell stimulated with LPS, and the inhibitory effects increased as the concentrations of indomethacin increased.

CONCLUSIONIndomethacin can inhibit the increase of IL-6 expression caused by supernatant of U937 cells stimulated with LPS in FLS.

Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Arthritis, Rheumatoid ; metabolism ; pathology ; Cells, Cultured ; Fibroblasts ; metabolism ; pathology ; Humans ; Indomethacin ; pharmacology ; Interleukin-6 ; biosynthesis ; genetics ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; genetics ; Synovial Membrane ; metabolism ; pathology ; U937 Cells

OBJECTIVETo compare the efficacy of different points combination in the treatment of menopausal insomnia.

METHODSNinety-six cases of menopausal insomnia were randomized into 3 groups, Xinshu (BL 15), Shenshu (BL 23), Sishencong (EX-HN 1), Shenmen (HT 7), Sanyinjiao (SP 6) were chosen in the restore interaction between the heart and the kidney group (group A, 32 cases); Zhaohai (KI 6), Jiaoxin (KI 8), Shenmai (BL 62), Pucan (BL 61) were chosen in the acupuncturing qiao mai group (group B, 32 cases); auricular Shenmen (TF4) and sensitive spot at the distribution area of auricular vagus nervus were chosen in the ear acupuncture group (group C, 32 cases). Six days made one session and the treatments were finished after 4 courses. The polysomnography (PSG) and Pittsburgh sleep quality index (PSQI) were employed before and after treatment to evaluate the alleviation of insomnia.

RESULTSThe parameters of the sleep latency (SL), rapid wave sleep latency (RL) and sleep efficiency (SE) were significantly improved in the three groups, and the differences were statistically significant (P < 0.05, P < 0.01). The SL and awaking time (AT) in group C [SL (401.08 +/- 16.54) min and AT (4.87 +/- 2.64) times] were significantly superior to those in the other two groups [SL (50.36 +/- 18.47) min, (54.87 +/- 20.92) min, AT (5.98 +/- 2.11) times, (6.13 +/- 3.04) times, all P < 0.05]. The S(3+4) (%) in group C was also significantly higher than those in the other two groups (both P < 0.05). It was indicated by PSQI that the sleep quality of group C (0.78 +/- 0.12) was significantly superior to that in group B (1.32 +/- 0.29), the total score and cured and markedly effective rate in group C [(4.34 +/- 1.43), 68.8% (22/32)] were superior to those in group A [(7.48 +/- 3.09), 53.1% (17/32), both P < 0.05].

CONCLUSIONEar acupuncture has a better curative effect than the restore interaction between the heart and the kidney group and acupuncturing qiao mai group, it is worth of being promoted.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Menopause ; psychology ; Middle Aged ; Sleep ; Sleep Initiation and Maintenance Disorders ; therapy ; Treatment Outcome