2. Optimization of urinary sample preparation process for real-time PCR detection of BK virus
Academic Journal of Second Military Medical University 2010;31(4):438-441
Objective: To develop an effective preparation method to improve the sensitivity and accuracy of real-time PCR in detection of BK virus's (BKV's) load in urine samples. Methods: A total of 24 samples documented as positive probes in primary detection were enrolled in this study. The candidate samples were prepared by 4 different approaches: unprocessed urine, BKV's DNA extracted from urine, 1:10 diluted urine, and 1:100 diluted urine; and then they were subjected to real-time PCR examination to obtain the viral load. The data obtained were analyzed by SPSS 11.0. Results: The four different preparation processes for urinary specimens had significant impact on detection results of real-time PCR. Three samples were negative in the unprocessed urine group and 66.7% of its samples had the lowest viral loads compared with the other three groups. Two samples in the 1:100 diluted urine group were negative and 79.2% of its samples had the highest viral loads, but its median load was similar to that of the 1:10 group. Viral gene was detected in all samples in the DNA extraction group and 1:10 diluted urine group, but the loss of the target gene was more severe in the DNA extraction group. Conclusion: The 1:10 diluted urine is better for real-time PCR detection of BKV's load, as it lose less viral gene and is more efficient, easy to perform and economical.
5.Blood glucose fluctuation and activation of oxidative stress in diabetes.
Chinese Journal of Pediatrics 2012;50(7):554-556
Blood Glucose
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metabolism
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Diabetes Complications
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prevention & control
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Diabetes Mellitus, Type 1
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blood
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metabolism
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physiopathology
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Diabetes Mellitus, Type 2
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blood
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metabolism
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physiopathology
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Dinoprost
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analogs & derivatives
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blood
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Glucose
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metabolism
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Glycated Hemoglobin A
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Humans
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Hypoglycemic Agents
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pharmacology
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Insulin
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pharmacology
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Oxidative Stress
6.Biochemical and Enzymatic Characterization of a Thermostable DNA Ligase Encoded by Thermophilic Acidophilic Archaebacterium Strain JP2
Haiyan LAN ; Chun LIU ; Phil HENDRY
Progress in Biochemistry and Biophysics 2006;33(9):881-889
A thermostable DNA ligase gene was identified, and the biochemical and enzymatic properties of the ligase were characterized from JP2 strain which was enriched from geothermally active sites in Papua New Guinea. The nucleotide and amino acid sequences showed much high identities compared with that of archeabacterium species Sulfolobus solfataricus and Sulfolobus shibatae,especially in the six conserved motif sequences, which are known to be closely related to the key function of ligase. Recombinant JP2 ligase showed high activity in nick-joining reaction. It was the most active when Mn2+ present as divalent metal cofactor rather than Mg2+ and Ca2+ etc.. Assay of thermostability over a range of temperatures showed that at 50~80℃ the enzyme displayed relative high activity. Further thermostability experiment indicated that the activity of JP2 ligase could last for a long time at 80℃ and 85℃,however, at 90℃ and 95℃, it became unstable quickly. An investigation on the acquired thermotolerance of recombinant JP2 ligase was done by applying a chaperonin known as TF55 in thermophile on JP2 ligase reaction. Result showed that TF55 could not help in improving thermostability of ligase at 85℃. The possible reason might be that at 85℃ in vitro, the chaperonin itself was denatured.
7.Effect of Rehabilitation Time on Functions of Patients with Intracerebral Hemorrhage
Hua-sheng PENG ; Chun-lan YUAN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(2):150-151
ObjectiveTo investigate the effect of rehabilitation time on the motor function and activities of daily living (ADL) of patients with intracerebral hemorrhage.Methods76 patients were randomly divided into the early rehabilitation group (40 cases) and late rehabilitation group (36 cases). The patients of two groups were treated with Bobaths method. The starting times of rehabilitation were 48 h~7 d (early rehabilitation group) and 3~4 weeks (late rehabilitation group) after onset. The therapeutic effect was assessed with Fugl-Meyer Motor Assessment Scale (FMA), modified Barthel index (MBI) and neurological deficit evaluation (ND).ResultsThe scores of the FMA, MBI and ND were not different between two groups before treatment. After one month treatment, the scores of patients in two groups improved significantly, but the early rehabilitation group had a greater improvement compared with the late rehabilitation group. Conclusion Early rehabilitation and late rehabilitation can also facilitate the motor function and ADL of patients with intracerebral hemorrhage recovery, and the therapeutic effect of early rehabilitation is better than late rehabilitation significantly.
8.Advance of the tumor therapy based on hypoxia-inducing factor inhibitors
China Oncology 2006;0(11):-
Hypoxia-inducible factor(HIF) plays a key role in the adaptation of tumour cells under hypoxic circumstance,there are some advances in the literatures regarding HIF as an important target for anticancer agents and gene therapy.In this review,the molecular mechanism and clinical significance of compounds targeting on hypoxia- inducing factor was summarized.
9.Comparison of infrared mrker-based positioning system and electronic portal imaging device for the measurement of setup errors
Yankun CAO ; Chao GAO ; Lan WANG ; Zifeng CHI ; Chun HAN
Chinese Journal of Radiation Oncology 2011;20(5):414-416
ObjectiveTo measure the setup errors with infrared marker-based positioning system (IM-BPS) and electronic portal imaging device (EPID) for patients with esophageal carcinoma and lung cancer and investigate the accuracy and practicality of IM-BPS. MethodsFrom January 2007 to January 2008, 40 patients with esophageal carcinoma and 27 patients with lung cancer received three-dimensional conformal radiotherapy or intensity-modulated radiotherapy, setup errors during the treatment were measured with IM-BPS and EPID, and the data of setup errors were compared with paired t-test and agreement with x2-test. ResultsIt takes 10 - 12 mins to complete the validating for each patient by EPID) system, while IMBPS system only needs 2 -5 mins. The mean setup errors along x, y and z-axis for patients with esophageal carcinoma measured by IM-BPS and EPID were 3.49 mm, 3. 19 mm, 3.31 mm and 4. 03 mm, 3.41 mm, 3.43 mm, respectively. For the patients with lung cancer, the setup errors were 4. 23 mm, 3.51 mm, 3. 39mm and 4. 85 mm, 3. 53 mm, 3.74 mm, respectively. The difference of setup errors meanured by the two systems was within 1 mm for 65% esophageal carcinoma patients ( x2 =51.09, P =0. 000), and 55% lung cancer patients ( x2 =53. 35, P =0. 000).Conclusions The measurement results of setup errors for patients with esophageal carcinoma and lung cancer show that IM-BPS is mostly better than EPID. Though validating for patients can be measured accurately and be well quality controlled, IM-BPS is used easily because of macroscopic, homely,spare time and real-time monitoring.