Detection of low-level somatic DNA mutations and minority alleles within a wild-type sample is crucial in fields of medicine,such as cancer,prenatal diagnosis and infectious diseases.Increasing enrichment methods have been developed to study this challenging area.They are typically segregate by their ability to enrich for,and detect,either known or unknown mutations,including COLD-PCR,ice-COLD-PCR,TT-PCR,PNA-mediated PCR,WTB-PCR and digital PCR.In this review,we discuss theoretical principles and relative advantages and disadvantages of these techniques,and put an emphasis on their applications in fields of diseases.

Objective To investigate the effect of different dose of GM1 on inducing human adipose tissue-derived stromal cells (ADSCs) into neural cells. Methods Human ADSCs were isolated from human liposuction tissues and cultured in NIM with different dose of GM1, which was used to induce ADSCs into neural cells. The experiment was separated into six groups: control group (?-MEM), Group A (NIM), group B (NIM+50 ?mol/L GM1), group C (NIM+100 ?mol/L GM1), group D (NIM+200 ?mol/L GM1) and group E (NIM+400 ?mol/L GM1). Cells' growth and morphologic changes were observed under invert phase-contrast microscopy. The expressions of Nestin, NSE, MAP2 and GFAP were identified by immunocytochemistry methods.Results (1)ADSCs could differentiate into neural cells in NIM with different dose of GM1. The number of neural cells differentiated from ADSCs increased in a certain period, especially in group E at each time point (all P

Insulin resistance and its compensatory hyperinsulinemia play a key pathogenic role in the infertility of the polycystic ovary syndrome.Numerous studies indicate that insulin sensitizing drugs can be used to enhance spontaneous ovulation and the induction ovulation in the syndrome,The aim of this review is to summarize the studies in which insulin sensitizing drugs were used to increase ovulation rate or improve fertility in women with the PCOS and to translate the information into practical guidelines for reproductive endocrinologist.

The α chain of hemoglobin of 615 mouse was isolated and purified on CM-Celullose-23 colomn chromatography. The N-terminal amino acid of the α chain was valine determined with DABITC/PITC method.The amino acid composition was determined and it was different from the parent(C57BL)in literature on the number of leucine residue,histine residue and valine residue.An undissoluble ‘core’ and dissoluble peptides were found when the α chain of 615 mouse was hydrolysised by trypsin and it was found that the eighth amino acid residue from N-terminal of one particular peptide fragment mutated from valine (C57BL) to leucine.

Objectives: Hypertensive disorders complicating pregnancy(HDCP) has a long-term deleterious effect.This study aimed to investigate the changes of cognitive function in women with HDCP and explore their possible mechanism.Methods: Fifty-three patients with HDCP and 22 normal pregnant women were included in this study,their cognitive functions assessed 60-80 days after delivery by word learning and delayed recall test,symbol digit substitution test,animal category fluency test,block design,trail making test(Part A) and digit span test.The results of the cognitive tests were compared between the HDCP and normal control groups.Results: Cognitive performances of the severe preeclampsia patients differed significantly from those of the normal individuals on the tests of word learning(P

Aim To investigate the effects of PLCε on the invasion and migration of human osteosarcoma cancer cells U2OS. Methods RNA interference ( RNAi) was used to inhibit PLCεexpression, and the proliferation of cancer cells was measured by CCK-8 assay. The migration of the cells was measured by scratch wound healing assay and migration chamber as-say. Gelatin zymography was performed to measure the MMP2 activities in U2OS cells. Results PLCε ex-pression was suppressed by siRNA. CCK-8 assay showed that PLCε had no effect on the proliferation of cancer cells. PLCε knockdown inhibited cell invasion and activities of MMP2 . Conclusion PLCε knock-down can inhibit the migration and invasion of human osteosarcoma cancer cells U2OS.

Objective To prepare ?-elemene solid lipid nanoparticles and investigate their particle size diameter and entrapment efficiency.Methods The ?-elemene solid lipid nanoparticles were prepared by film ultrasonic wave dissolving techniques.And the optimum formula was selected through orthogonal design test according to the entrapment efficiency.Results The optimizing technique was ?-elemene(20 ?L),stearic acid(90 mg),lecithin(90 mg),Tween80(2.5 %,5 mL) and Poloxamer 188(2.5 %,5 mL).Conclusion The technique of preparing ?-elemene by film-ultrasonic wave dissolving technique is feasible.

Objective: To observe the changes of the cognitive function of patients with polycystic ovary syndrome(PCOS) and explore their relationship with serum testosterone.Methods: The levels of serum testosterone(T),dehydroepiandrosterone sulfate(DHEAS),sex hormone binding globulin(SHBG),Estradiol(E2),follicle-stimulating hormone(FSH) and luteotrophic hormone(LH) were measured by radioimmunity assay in 25 patients with PCOS and 25 normal women.Their cognitive functions were assessed by delayed word recall test,word learning test,symbol digit substitution test,animals category fluency test,block design,trail making test(Part A) and digit span test.And the results of the tests were compared between the PCOS and the normal group.Results: The levels of serum T,LH and SHBG in the PCOS patients differed significantly from the normal individuals(P

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis, however, its mechanism has not been fuly elucidated. Urokinase type plasminogen activator system which participated in the degradation of the extracelular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE: To determine the effect ofTougu Xiaotong capsule on urokinase-type plasminogen activator system in knee cartilage tissues of knee osteoarthritis rats. METHODS: Of 144 Sprague-Dawley rats, 120 rats were randomly made into models of knee osteoarthritisvia intra-articular injection of papain, and randomly assigned to model group,Zhuanggu Guanjie Wan group [1.2 g/(kg?d)], low-doseTougu Xiaotong capsule group [0.092 g/(kg?d)], moderate-doseTougu Xiaotong capsule group [0.184 g/(kg?d)] and high-doseTougu Xiaotong capsule group [0.368 g/(kg?d)]. Each group contained 24 rats. Every 2 weeks was considered as a course, with a 2-day interval, totaly 4 courses. The remaining 24 normal rats were included in the blank group. After every two courses, a batch of experimental animals was sacrificed. The pathological changes were observed folowing staining with hematoxylin and eosin. The positive cels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by immunohistochemistry. The protein levels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by western blot assay. RESULTS AND CONCLUSION:Mankin’s score was significantly lower in theTougu Xiaotong capsule group and Zhuanggu Guanjie Wan group compared with the model group (P < 0.01), in a time-dependent manner. Immunohistochemical staining indicated that the positive cels of urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor were significantly decreased, but plasminogen activator inhibitor was significantly increased in theTougu Xiaotong capsule group andZhuanggu Guanjie Wangroup in a time-dependent manner. Western blot assay results had an identical trend to immunohistochemistry. These indicated thatTougu Xiaotong capsule showed preventive and therapeutic effects on osteoarthritis by regulating urokinase-type plasminogen activator system.

BACKGROUND:Tougu Xiaotong Capsule has pretty good clinical therapeutic effect on osteoarthritis of early and middle periods. However, the mechanism of Tougu Xiaotong Capsule is not ful y clarified. The RhoA GTPases can regulate chondrocyte apoptosis and hypertrophy.
OBJECTIVE:To observe the Tougu Xiaotong Capsule on the expression of Rac1and Cdc42 in tumor necrosis factor-α-induced in vitro cultured rat articular chondrocytes, and to explore its mechanism of action for combating osteoarthritis.
METHODS:Knee cartilage of the 4-week-old Sprague-Dawley rats was used to stably establish in vitro culture system of chondrocytes. Passage 3 chondrocytes were identified by toluidine blue staining. Chondrocyte apoptosis was successful y induced by 20μg/L tumor necrosis factor-αand then Tougu Xiaotong Capsule at different dosage (500, 100, 20 mg/L) was given after 24-hour incubation. MTT assay was used to detect cellsurvival, flow cytrometry to measure mitochondrial membrane potential, and western blot assay to determine the protein expression of Rac1, Cdc42, Bax and Bcl-2.
RESULTS AND CONCLUSION:Tougu Xiaotong Capsule could reduce tumor necrosis factor-α-induced apoptosis of chondrocytes to improve the survival rate of the cells, and at the same time, could down-regulate the protein expression of Rac1, Cdc42 and Bax and increase the protein expression of Bcl-2 significantly (P<0.05). Tougu Xiaotong Capsule possibly plays a therapeutic efficacy on osteoarthritis by reducing promote apoptosis Rac1, Cdc42 and Bax expression and increasing apoptosis inhibiting gene Bcl-2 expression, thereby to inhibit apoptosis of chondrocytes.