Recently,much interest has been focused on the clinical application and biology of tumor-derived DNA in the plasma/serum of cancer patients.Such interest has resulted in the demonstration of human papillomavirus(HPV) DNA in the plasma/serum of patients with cervical cancer(CC).The studies used different materials(plasma /serum),different DNA extraction methods and different primers to detect circulating free HPV DNA,so the positive rates varied from(6.9%) to 50%.The HPV DNA in the plasma/serum of patients suffering from CC most likely originates from the tumor itself,which will probably provide us with a new tool for CC monitoring after primary treatment,especially in patients in advanced stages.It is expected that this promising molecular tumor marker would soon be put into routine clinical use.

Background and purpose:For the past few years, HPV DNA has been detected in the peripheral blood in the patients with cervical cancer and believed to be a promising tumor marker, but because the concentration in the patients' serum was extremely low, it made it difficult to detect the circulating HPV DNA. In order to find a way to improve the sensitivity of the assay, this study compared and evaluated three methods that are currently being used to measure HPV DNA in the extraction from serum of cervical cancer patients. Methods:51 patients were pathologically proved to be cervical cancer and enrolled into the study. DNA from their sera was extracted by three methods:(1) Phenol-chloroform extraction, (2) UNIQ-10 collumn virus DNA kit(SANGON,Shanghai), (3) QIAamp MinElute Virus spin kit(QIAGEN,Germany). Then the HPV DNA was quantitated by PCR using GP5+/GP6+ primers.Results:Either fresh or short-term storage sera has been used The positive rates were 14.3%, 5.7%, and 57.1% with three assays, respectively. But when long-term storage samples were used to quantitate, the positive rates were low regardless of the methods being used.Conclusions:QIAamp MinElute Virus spin kit was the most sensitive for the content of serum HPV DNA, it could improve the positive rate for the assay if the serum was not stored for long time.

To investigate the changes of plasma adrenomedullin(ADM) and adrenotensin(ADT) levels and their clinical significance in the pathological process of congestive heart failure(CHF), plasma levels of ADM and ADT in 45 patients with CHF before and after treatment were measured by specific radioimmunoassay. The results indicated that before treatment, plasma levels of ADM in class Ⅱ and Ⅲ CHF patients were 51.46?4.52pg/ml, 70.39?3.22pg/ml and plasma levels of ADT in class Ⅱ and Ⅲ CHF patients were 29.98?1.13pg/ml, 33.45?0.91pg/ml, respectively,which were higher than those of the control subjects(24.12?1.59pg/ml and 24.89?2.19pg/ml, respectively)(P

Objectives To study the changes of plasma levels of proadrenomedullin N terminal peptide(PAMP) in patients with congestive heart faillure(CHF) before and after drug treatment and its significance.Methods Plasma PAMP levels in 45 patients with CHF and 7 heathy control subjects were measured by specific radioimmunoassy.Results The plasma PAMP levels in patients with CHF were significantly decreased with the deterioration of cardiac function.Plasma PAMP levels in 45 patients in NYHA class Ⅳ(2 79?0 89pg/mL) were significantly lower than those in class Ⅱ(6 24?1 71pg/mL)?class Ⅲ(7 38?1 28pg/mL) and control subjects(8 56?2 44pg/mL)(P

Objective To investgate role of TLR2 in the activation, the innate immune and inflammation of human platelets. Methods Human washed platelets were separated from healthy people(n=5) and were stimulated with different concentrations(1μg/ml, 5μg/ml, 10μg/ml) of TLR2 agonistPam3CSK4(a synthetic bacterial lipoproteins). Then the platelet aggregation rate, the expression of CD62p and TLR2 on the platelet surface were measured. Results The platelet aggregation rate were (28.32±5.67)%, (52.56±8.54)% and (76.24±11.23)%, respectively, at concentration of 1μg/ml, 5μg/mland 10μg/ml of Pam3CSK4, more than (12.83±2.43)% at 0μg/ml of it. In addition, the expression of CD62p were (18.45±2.66)%, (22.45±2.04)%, (29.53±4.08)%, respectively at above concentration of Pam3 CSK4, more than (11.20±1.67)% of CD62p at control group(P<0.01). The expression of TLR2 was not significantly increased at a lower concentration of Pam3CSK4(1μg/ml) with (16.85±6.10)% compared with(10.81±3.99)% at the control group. However, it were (21.15±9.90)% and (22.52±9.26)%, respectively, at a higher concentration(5μg/ml, 10μg/ml)more than(10.81±3.99)% at the control group(P<0.05). Conclusion Pam3CSK4 induce aggregation, activation and the up-regulation of TLR2 of platelet by stimulating TLR2 receptor of it. Thereafter, TLR2 play an important role in the innate immune of platelet.

Objective To evaluate the importance of factors affecting efficiency of image display quality of realtime three-dimensional echocardiography (RT-3DE) and establish an optimized method for RT-3DE examination and image processing in children. Methods Based on corresponding cardiac acoustic characteristics in children, 87 healthy children (46 boys, 41 girls, mean 36.2 ± 44.2 months) were selected. Three aspects of seven factors were selected, including acquisition windows, gain, compress, post process, smoothing, frequency fusion and 3D vision for RT-3DE examination and analysis using orthogonal test design method. The principal effectiveness analysis of orthngonal test design method and its table L_(18)3~7 were used. The efficiency rate of 3D image display quality was used as the inspection index for orthogonal test. Results According to the average deviation orders of the seven factors, the maximal average deviation was gain. The aspects with maximal average deviation of each factor were at via-subcostal and via-apical acoustic windows, 90 of gain, 60 of compress, E of post process, 5 of smoothing, F3 of frequency fusion and A to B of 3D vision, respectively. Conclusions The optimized method for RT-3DE examination and image processing could be achieved by setting acquisition windows, gain, compress, post process, etc. It is helpful to promote clinical application of RT-3DE and benefits the design of multi-parameter presetting of ultrasound system to optimize RT-3DE examination in children.

BACKGROUND:The endothelial dysfunction is the pathogenesis of arteriosclerotic disease, the quantity and function of endothelial progenitor cells are decreased within the cycle, leading to a poor capacity of neovascularizatio, the efficacy of stem celltransplantation alone is unclear, the combination of cytokines and gene-modified stem cells is the hotspot.
OBJECTIVE:To observe the effect of stromal cel-derived factor-1 on the neovascularization after endothelial progenitor cells transplantation.
METHODS:Unilateral hindlimb ischemia model was established in 20 athymic nude mice, and the mice were randomly divided into four groups:combined group (intravenous endothelial progenitor cells+intramuscular stromal cel-derived factor-1), endothelial progenitor cells group (intravenous injection of endothelial progenitor cells), stromal cel-derived factor-1 group (intramuscular injection of stromal cel-derived factor-1), and blank control group (intramuscular M199). The skin temperature of ischemic hindlimbs and survival of animals after transplantation were observed. The ratio of capil ary/skeletal muscle fiber was counted. The expression of CD31 and endothelial nitric oxide synthase were detected.
RESULTS AND CONCLUSION:The fluorescence-labeled endothelial cells were embedded in ischemic hindlimb muscles after celltransplantation. Of the 20 nude mice, two mice died. The rate of ischemic hindlimb reserving was respectively 80%, 75%, 20%and 0 in combined group, endothelial progenitor cells group, stromal cel-derived factor-1 group, and blank control group. The capil ary/muscle fiber ratio in combined group and endothelial progenitor cells group was higher than that of blank control group (P<0.01). The combined group was greater than endothelial progenitor cells group, and endothelial progenitor cells group was greater than stromal cel-derived factor-1 group (P<0.05). The capil ary density in combined group and endothelial progenitor cells group were higher than that in blank control group (P<0.01), and stromal cel-derived factor-1 group was also more than blank control group (P<0.05). The combined group was greater than endothelial progenitor cells group, and endothelial progenitor cells group was greater than stromal cel-derived factor-1 group (P<0.05). The positive rate of endothelial nitric oxide synthase was 73.33%and 53.33%in combined group and endothelial progenitor cells group respectively (P>0.05). Endothelial progenitor cells can migrate to ischemic tissues, endothelial progenitor cells transplantation can promote neovascularization, and stromal cel-derived factor-1 augments the neovascularization after celltransplantation, in which endothelial nitric oxide synthase is involved.

Objective To discuss the technology of modest isolating and enriching the intestinal epithelial stem cells. Methods Mouse intestinal epithelial cells were stained by Rhodamine 123 (Rho), sorting the Rhodamine 123 low staining cell population ( Rholow ) and Rhodamine 123 strong straining cell population ( Rhobri ) by fluorescence activated cell sorting(FACS) in flow cytometer; Detecting the musaashi-1 and p-glycoprotein 1 (p-g-p1) mRNA expression of two groups by RT-PCR; Analyzing the cell cycle and the percentage of the musashi-1 positive cells by flow cytometry. Results The intestinal epithelial cells were divided into three groups, Rhodamine 123 low staining cells( 12. 34% of total cells), Rhodamine 123 middle staining cells (45.26% of total cells) and Rhodamine 123 strong staining cells ( 41. 40% of total cells). The Rholow cell fraction and Rhobri cell fraction were isolated successfully. Both of musashi-1 and p-g-p1 mRNA were strongly expressed in Rho1ow cell fraction, and Rhobri cell fraction little expressed p-g-p1 mRNA. Most of Rho1ow cells were in G0/G1 phase, and the musashi-1 positive cells were about 10.37% of total cells in this fraction. Conclusion The intestinal epithelial stem cells can be modestly isolated and enriched by Rhodamine 123 staining.

Objective To observe the clinical efficacy of bamboo-circled salt-partitioned moxibustion in treating arthritis of temporomandibular joint.Method Eighty patients were randomized into two groups. Forty cases in the bamboo-circled salt-partitioned moxibustion group received bamboo-circled salt-partitioned moxibustion at temporomandibular joint; forty cases in the warm needling group were intervened by selecting Xiaguan (ST7), Ashi point, etc. at the affected side. For the two groups, 3-day treatment was taken as a treatment course, and the therapeutic efficacy was analyzed after 2 treatment courses. The improvements in pain and mouth opening were observed before and after the treatment, and the treatment efficacy was evaluated by a 1-month follow-up study.Result The bamboo-circled salt-partitioned moxibustion group was superior to the warm needling group in comparing the real-time analgesic effect (P<0.05) and in the improvement of mouth opening (P<0.05); the comprehensive markedly effective rate was respectively 67.5% and 45.0% in the bamboo-circled salt-partitioned moxibustion group and warm needling group, and the between-group difference was statistically significant (P<0.05), indicating that bamboo-circled salt-partitioned moxibustion is better than warm needling in treating arthritis of temporomandibular joint; the follow-up study revealed satisfactory therapeutic efficacies in both groups: the effective rate was 92.5% in the bamboo-circled salt-partitioned moxibustion group versus 87.5% in the warm needling group, and the difference was statistically insignificant (P>0.05).Conclusion Bamboo-circled salt-partitioned moxibustion can produce a real-time analgesic effect and improve mouth opening; it's especially suitable to treat the patients who are afraid of needling, as it's significantly effective, safe, non-invasive,and easy-to-operate.

Objective:To investigate effects of over-expression and suppression of HMGB1 on proliferation and invasion of endometrial carcinoma HEC-1A cell and underlying mechanisms.Methods:Over-expression or silence of HMGB 1 in HEC-1A cell lines were established by lentiviral vector containing HMGB 1 recombinant plasmid or by HMGB 1 shRNA,respectively.Cell counting kit-8,Transwell,and wound healing assay were used to analyze proliferation,invasion,and migration of HEC-1A cells,respectively.Western blot and reverse transcription-PCR (RT-PCR) were used to detect the expression of NF-κB,VEGF,and matrix metalloproteinase 2 (MMP2) in the cells.Results:Over-expression of HMGB1 promoted the proliferation,invasion,and migration of HEC1A cell,and up-regulated NF-κB,VEGF,and MMP2 expressions,while suppression of HMGB1 inhibited the proliferation,invasion,and migration of HEC-1A cells and down-regulated NF-κB,VEGF,and MMP2 expressions.Conclusion:HMGB1 plays an important role in proliferation,invasion,and migration of HEC1A cell via NF-κB/VEGF/MMP2 pathway.HMGB 1 might be a potential target for endometrial carcinoma therapy.