Objective To develop a nested-real-time PCR method for the detection of FOXP3 mRNA in human peripheral blood mononuclear cells(PBMC).Methods The outer primers were designed by span intron,and the inner primers and TaqMan probe were designed according to the amplified sequence.During the nested real-time PCR,decrease primers concentration and cycle times were decreased in first PCR amplification to increase specific templates,then quantitative real-time fluorescence was detected in the second PCR amplification.The standard curve was created by FOXP3 and beta-actin recombined plasmid DNA respectively,and the results were presented as the ratios of FOXP3 mRNA to beta-actin mRNA.Results Total mean Ct(threshold cycle) in 6 tests was 14.66.The mean CV(coefficient of variation) was 5.81%.The standard curve showed a fine linear relationship between Ct and template concentration,and the correlation coefficient was 0.999.The sensitivity was 100 times higher than normal real-time PCR by reach 100 copy.Conclusions Nested-real-time-PCR for the detection of FOXP3 mRNA in human PBMC is a simple,repeatable,high specific and sensitive method to evaluate immune function and observe therapeutic effects in certain diseases for clinical routine application.

Objective To establish a nested-real-time-PCR for the detection of IL-21R mRNA in peripheral blood mononuclear cells(PBMC)of patients with rheumatoid arthritis,and explore its clinical significance.Methods The outer primers of span intron and inner primers were designed for nested-real-time-PCR.To increase specific template,the concentration of primers and enzyme,cycle times were decreased in the first amplification,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of IL-21R mRNA to ?-actin mRNA.Results IL-21R mRNA was detected in 60 patients with RA and 30 healthy controls.The expression of IL-21R mRNA gene in the patients with RA was increased,and the statistical analysis has significant differences(P0.05),but a significant difference was found in the patients of grade III compared with those of grade I and II(P

Objective To detect the ratio of CD4+ and CD8+ T cells and the number of CD4+CD25+ T regulatory cells(Treg)in human peripheral blood mononuclear cells(PBMC)of rheumatoid arthritis(RA)patients and evaluate the significance for diagnosis of RA.Methods The number of CD4+,CD8+ T cells and Treg cells were detected by flow cytometry and the ratio of CD4/CD8 was calculated.Results The ratio of CD4/CD8 and the number of CD4+CD25+ T cells detected in 49 RA patients were significantly higher than that in health group with statistical significance(P

Objective To develop a nested-real-time fluorescence quantitative PCR method for detection of TGF-?RII mRNA in human peripheral blood mononuclear cells(PBMC),and analyze the clinical implications of TGF-? RII in RA patients.Method The outer primers of span intron and inner primers were designed for nested-real-time fluorescence quantitative PCR.To increase specific template,the concentration of primers and enzyme,cycle times in the first amplification were decreased,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of TGF-? RII mRNA to ?-actin mRNA.Results The standard curve showed a fine linear relationship between Ct(cycle threshold)and logarithm of template concentration,and the correlation coefficient was 0.999.The sensitivity reached 101 copies.The TGF-? RII gene expression in PBMC of RA patient was increased compared to healthy controls[(0.45?0.11)vs(0.37?0.10),P

Objective To investigate the expression of genes associated with TGF-?/Smad signaling pathways in peripheral blood mononuclear cells(PBMC)of rheumatoid arthritis(RA)patients and to evaluate the relevant clinical significance.Methods Conventional and nested SYBR green I fluorescent dye real-time PCR was used to detect the expression of TGF-? RII,Smad4,and Smad7 mRNA.Results Compared with those of 29 healthy persons,the expressions of TGF-? RII and Smad4 mRNA,and the ratio of Smad4/Smad7 in PBMC of 58 RA patients were significantly increased (P

Objective To evaluate the effectiveness of a health education program based on the three level maternal and child health care networks in terms of acceptability of HIV tests in pregnant women. Methods A field study approach was adopted. Pregnant women in two sites who were similar in their social features were selected as intervention group and control group,respectively.Based on the baseline survey,the intervention group was exposed to one-year long health education activities on prevention of HIV related MTCT via lectures,broadcasting and educational materials etc.The effectiveness of the interventions was evaluated by interviewing the pregnant women in the two groups via questionnaires and comparing change of HIV test rates pre-and post-interventions. Results A total of 1 541 pregnant women were interviewed,with 933 women in the intervention group and 618 women in the control group.After one year intervention,the rate of HIV-testing during pregnancy in the intervention group was raised from 34.27% to 72.49%,while 5.83% to 20.38% in the control group.The rate of HIV-testing during pregnancy in the intervention group was 52.11% higher than in the control group (?2=24.60,P

Objective To analyze the causes of surgical errors and complications of lumbar degenerative diseases treated by transforaminal endoscopic spine system(TESSYS),and explore the treatment and prevention measures.Methods From July 2012 to January 2016,the data of 660 patients with lumbar degenerative diseases who treated with TESSYS were analyzed retrospectively,of which 528 patients were lumbar disc herniation with 192 cases of single segment(L4/L5) and 336 cases of multi-segment.Lumbar spinal stenosis in 132 cases,were lateral recess and nerve root canal stenosis.Surgical errors and complications were recorded,and analyzed the reasons and the treatment and preventive measures.Results Intraoperative errors:1 case of sterile film with the dilating catheter went into the surgical area;1 case with wire fracture,and stump retention in vivo;for discography staining,the proportion of mixed solution was not enough,the leakage of excess concentration of methylene blue caused caudal nerve injury in 3 cases;expansion of the intervertebral foramen,the drilling part of the articular process was too deep and too close to the medial wall of the articular process,which caused cutting injury of nerve fiber sheath in 2 cases;after the rupture of the dura mater,the rinse solution was perfused,caused bloating,elevated intracranial pressure,elevated blood pressure and other symptoms of spinal cord hypertension in 3 cases;low dose of local anesthetic into the blood caused toxic reaction in 2 cases;total spinal cord anesthesia with paravertebral nerve root block in 1 case.Postoperative complications:3 cases of nerve root edema;2 cases of free nucleus pulposus tissue;2 cases of local hematoma led to nerve root compression;1 case of intervertebral space infection and 3 cases of local skin neuritis in the puncture area.Conclusion In the process of intervertebral foramen surgery,there are some deficiency on instruments or operating skill.Therefore,rigorous treatment of each link in the perioperative period,is the guarantee of successful surgery.

Objective To observe the effect of comprehensive community intervention on cardiovascular risk factors in metabolic syndrome (MS) high risk group.Methods One hundred MS high risk patients were divided into intervention group (50 patients) and control group (50 patients) by random digits table.The intervention group accepted intervention on health education,health action,health habit and individual intervention on drug.The control group accepted natural intervention.The changes of waistcircumference,body mass index (BMI),blood pressure,fasting plasma glucose (FPG),lipids and serum uric acid (UA) were observed.Results In intervention group,the number of physical exercise,alimentary control and medication compliance increased 12％,16％ and 18％,and the number of smoking and alcohol drinking decreased 8％ and 6％.But in control group,the number of physical exercise,alimentary control,medication compliance increased 2％,0 and 4％,and the number of smoking and alcohol drinking decreased 4％ and 2％.There was significant difference (P＜ 0.05).Before treatment,the level of waist circumference,BMI,systolic pressure,diastolic pressure,FPG,total cholesterol,triacylglycerol,high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C) and UA between two groups had no significant difference (P ＞ 0.05).After 1 year's intervention,except HDL-C,the level of other index had significant difference between two groups (P ＜ 0.05 or ＜ 0.01).Conclusion Comprehensive community intervention can improve the status of obesity and dyslipidemia,and reduce blood pressure,plasma glucose and UA.

Hypertrophic cardiomyopathy (HCM) is featured by asymmetry myocardial hypertrophy ,mainly involves interventricular septum ,and its clinical manifestations are left ventricular dysfunction and arrhythmia .It′s thera-peutic methods include medication ,intervention ,surgery ,dual chamber pacemaker (DDD) treatment and implant-able cardioverter /defibrillator (ICD) treatment etc .This article made following overview on treatment progress of HCM .

To explore the mechanism of Notch in hyperoxia-induced preterm rat lung injury, 2-days-old preterm SD rats were randomized into control and hyperoxia group (FiO2 > or = 0.85). On day 1, 7, 14 and 21, 8 rat pups of each time point were used to assess histopathological changes of lung with HE staining and to evaluate the expression of Notch1 and Notch3 with immunohistochemistry. Notch1, Notch3, Aquaprin5 (AQP5) and surfactant protein C (SP-C) mRNA were measured by reverse transcription polymerase chain reaction (RT-PCR). The results showed that the lung injury in the hyperoxia group was characterized by retarded lung alveolization and differentiation of alveolar epithelial type II cells (AEC II). Positive staining of Notch1 in hyperoxia group was weaker than controls at every time point (except for day 7), while positive staining of Notch3 was much stronger (P < 0.05, P < 0.01). Notch1, Notch3 mRNA level showed similar change as protein level. AQP5, SP-C mRNA decreased significantly as compared with that of the controls (P < 0.01). We are led to conclude that hyperoxia results in abnormal expression of Notch, which is likely to contribute to the pathogenesis of lung injury through regulating proliferation and transdifferentiation of alveolar epithelial cells.
Aerobiosis ; Animals, Newborn ; Lung/*pathology ; Lung Diseases/etiology ; Lung Diseases/*metabolism ; Lung Diseases/pathology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Random Allocation ; Rats, Sprague-Dawley ; Receptors, Notch/*biosynthesis ; Receptors, Notch/genetics