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 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 investigate the antiatherosclerotic effect of Wujibaifeng Pills and its mechanism. Methods: Rat dyslipidema was induced by ovariectomy (OVX), the levels of serum lipids, superoxide dismutase (SOD) activities and malondialdehyde (MDA) were detected after 12 weeks with the treatment of wujibafeng pills. Results: Wujibaifeng Pills lowered serum oxidized-low density lipoprotein cholesterol (OX-LDL,P

Objective To use metabonomics method to study the change of the basic materials of month rhythm of wei qi deficiency syndrome; To find the potential markers so as to provides a new way for the essence of the wei qi deficiency syndrome research.Methods Based on the autumnal equinox in lunar calendar month, the beginning of a month (the first day of lunar August), the middle of a month (the 15th day of lunar August), and the end of a month (the 30th day of lunar August) were set as the three days to draw experimental materials. Two weeks before drawing materials, 20 rats were randomly divided into control group and model group, 10 rats in each group. The model rats were modeled by the stimulus of fatigue combined with coldness and hotness. Control group rats received conventional breeding. The rats in the both groups during the three experiments received decollation and the blood was taken at the 12 o’clock at noon. HPLC-MS was used to detect plasma metabolites, and partial least squares were used to make statistical analysis on the data for comparing plasma metabonomics original data of control group and model group. Possible metabolic markers of wei qi deficiency syndrome were explored, and the potential makers of month rhythm change of wei qi deficiency syndrome were deduced.Results Oleamide, phosphatidyl glycerol, cortisol, proline, dimethyl fumarate, and eicosapentaenoic acid may be potential markers of wei qi deficiency syndrome in the beginning of a month. Sphingosine-1-phosphate, malic acid, cortisol, oleamide, carnitine, eicosapentaenoic acid and dimethyl fumarate may be potential markers of wei qi deficiency syndrome in the middle of a month. Cholesteryl acetate, threonine, cortisol, dimethyl fumarate, oleamide, eicosapentaenoic acid and pyroglutamate may be potential markers of wei qi deficiency syndrome in the end of a month.Conclusion Month rhythm change of wei qi deficiency syndrome may be influenced by oleamide, cortisol, eicosapentaenoic acid, dimethyl fumarate, and aconitic acid, and may be closely related to energy metabolism, meanwhile accompanied by regulation of cell, hormone and nerves.

Objective To investigate antimicrobial resistance of Escherichia coli (E.coli)and Klebsiella pneumoniae (K.pneumoniae),antimicrobial use density(AUD),as well as relation between antimicrobial resistance and AUD in a ter-tiary first-class hospital.Methods Antimicrobial resistance rates of clinically-isolated E.coli and K.pneumoniae,AUD of carbapenems and quinolones,as well as relation between resistance and AUD in 2013-2015 were statistically analyzed. Results Correlation analysis of antimicrobial resistance of bacteria and AUD showed that the decrease in resistance rate of E.coli to levofloxacin was related to the decrease in the use density of quinolones(r=0.61,P=0.03);increase in resist-ance rate of K.pneumoniae to imipenem was related to the increase in the use density of carbapenems(r=0.78,P<0.01). Conclusion Antimicrobial use is one of the causes of bacterial resistance,management on antimicrobial use needs to be strengthened to reduce the threat of bacterial resistance to human health.

Objective To study the plasma metabolomics features of SD rats with wei-qi deficiency respectively in spring, summer, autumn and winter; To seek the potential markers related to wei-qi deficiency; To discuss the essence of wei-qi deficiency.MethodsTwenty male SD rats were adaptively fed 7 days before Chinese Vernal Equinox Day, Summer Solstice, Autumn Equinox and Winter Solstice, synchronous controled with temporal condition. The rats were randomly divided into experimental and control groups according to random number method. Rats in the experimental groupwere made into wei-qi deficiency models through fatigue stimulation alternately with cold and heat stimulation. Blood was collected at 12:00 each solar term. High performance liquid chromatography- mass spectrometry instrument was used to detect plasma metabolites. Partial least squates-discriminant analysis was used to process statistical analysis of the data to compared the original data of plasma metabolomics between the two groups of each season, and explore the difference metabolic markers between each two groups, then speculate the potential biomakers of wei-qi differency.ResultsWith the changes of spring, summer, autumn and winter, significant differences were showed in metabolic profile between the experimental and control groups. Homocysteine showed a significant difference in spring, summer and autumn; Ceramide showed a significant difference in summer and winter; Testosterone showed a significant difference in spring and summer; Cyclic guanosine monophosphate showed a significant difference in autumn and winter; Sarcosine showed a significant difference in spring, summer and winter. The physiological functions of these substances related to lipid metabolism, amino acid metabolism, neurotransmitter and hormone regulation.ConclusionPotential biomakers of wei-qi deficiency changed along with the seasonal variation, which mainly reflected in lipid metabolism, amino acid metabolism, neurotransmitter and hormone regulation.

OBJECTIVE: To investigate the effect of neoadjuvant chemotherapy on cervical carcinoma and its association with clinical data. METHODS: A total of 97 patients with stage Ib2 approximately IIIa of cervical cancinoma were treated with neoadjuvant chemotherapy. The effect of chemotherapy, factors associated with outcome of chemotherapy, and histology were analyzed. RESULTS: Effective rate of chemotherapy was 86.6% which was associated with clinical stage and histology. Eight-four patients received radical hysterectomy. The histological grade of 17 patients was lowered, lymph nodes in 19 patients were positive, and 6 patients had parametrium invasion. One patient died within 1 year after the operation, and 5 patients recurred. CONCLUSION The effect of neoadjuvant chemotherapy for locally advanced cervical cancinoma is good. Surgery after chemotherapy can improve the prognosis and 5-year survival rate.
Adenocarcinoma ; drug therapy ; mortality ; surgery ; Adult ; Carcinoma, Squamous Cell ; drug therapy ; mortality ; surgery ; Female ; Humans ; Middle Aged ; Neoadjuvant Therapy ; methods ; Prognosis ; Survival Rate ; Uterine Cervical Neoplasms ; drug therapy ; mortality ; surgery

OBJECTIVE: To investigate the prevalence, etiology, clinical presentation and pathological features, treatment and prognosis of cervical cancer in young women. METHODS: Clinical data of 132 young women with cervical cancer were reviewed. RESULTS: Positive rate of human papillomavirus 18 was high in young women with cervical cancer. The primary clinical presentation of young patients with cervical cancer was contact bleeding of vagina, and the signs were out-expanding of cervical mass. The percentage of adenocarcinoma increased. The main treatment for cervical cancer was surgery. The patients had radical hysterectomy plus ovarian transplantation, none of whom had ovarian metastases and menopause syndrome. Prognosis of most patients was good. CONCLUSION Contact bleeding is a significant symptom in young women with cervical cancer. Surgery is first considered in the treatment. Preoperative neoadjuvant chemotherapy can be used in patients with locally advanced and late stage cervical cancer. Ovarian transplantation during operation can retain the ovary function.
Adult ; Age Factors ; Female ; Human papillomavirus 18 ; isolation & purification ; Humans ; Papillomavirus Infections ; complications ; Prognosis ; Uterine Cervical Neoplasms ; diagnosis ; surgery ; therapy ; virology ; Young Adult

Objective To explore the effect of drug injection under bronchoscopy on the retreatment of smear positive cavitary pulmonary tuberculosis. Methods From June 2016 to December 2017,164 cases of pul-monary tuberculosis with smear Yang cavity type were selected,which were divided into 2 groups according to the random digital table method,each groups has 82 cases.The control group received routine treatment,the observa-tion group underwent bronchoscopy with Kangfuxin Liquid combined with drug injection therapy.The clinical effect of the two groups,the changes of lung function before and after treatment and the improvement of clinical symp-toms were compared.Results The clinical curative effect of the treatment group was better than the control group, which difference was statistically significant(P < 0.05). FEV1,FEV1/FVC,PEF index in the observation group were better than the control group,which difference was statistically significant(P<0.05).The complication rate of observation group was lower than the control group,which difference was statistically significant(P < 0.05). Conclusion Drug injection under bronchoscopy can significantly improve the clinical efficacy and lung function and promote clinical symptoms in patients with retreated smear positive pulmonary tuberculosis.It is worthy of popu-larization and application.

OBJECTIVE: To study the proteins related to paclitaxel-resistant of ovarian cancer cell line. METHODS: The total proteins of paclitaxel-resistant and paclitaxel-sensitive human ovarian cancer cell lines were separated by 2-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed using image analysis software. The differential proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot was used to determine the differential expression levels of the 2 proteins. RESULTS: Forty differentially expressed proteins were found by image analysis software, and 24 differential proteins were identified by mass spectrometry. These proteins included proliferation cell nuclear antigen (PCNA), nm23, prohibitin (PHB), annexin, alpha-enolase, heat shock protein (HSP), and so on. CONCLUSION Twenty-four proteins in human ovarian cancer cell lines of paclitaxel-resistant and paclitaxel-sensitive are found by proteomic techniques, which may be involved in the paclitaxel-resistance of human ovarian cancer cells.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; NM23 Nucleoside Diphosphate Kinases ; analysis ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Paclitaxel ; pharmacology ; Proliferating Cell Nuclear Antigen ; analysis ; Proteome ; analysis ; Proteomics ; methods ; Repressor Proteins ; analysis