Objective To develop and verify a multiplex fluorescence quantitative PCR(Taqman probe) method for the detection of telomerase activity.Methods Specific reverse transcription primers,two pairs of quantitative primers and probes were designed for the CDS sequence of telomerase catalytic subunit telomerase reverse transcriptase(TERT).After optimization of the reverse transcription primers(specific reverse transcription primers and random primers) and quantitative primers(two pairs of quantitative primer probes used alone or in combination) in the reaction system,with the primer probe of internal reference gene GAPDH,multiplex fluorescence quantitative PCR was performed in a single tube.In addition,telomerase positive standard and negative standard were prepared with 293T and MRC-5 cells respectively,and the stability and precision of the method were verified.The telomerase activity in 19 normal mesenchymal cell samples and 32 breast cancer cell samples were detected by the developed method.Results The optimum reaction system was as follows:using cDNA synthesized with specific reverse transcription primers as the template,2 pairs of quantitative primer probes of TERT gene were mixed with internal reference gene GAPDH primer probes for multiplex fluorescence quantitative PCR reaction in a single tube.After optimization,the sensitivity and TERT fluorescence signal quantity of the system were greatly improved,and the ΔRn was enhanced by 3 times.The amplification curve of positive standard TERT gene was normal,and the ΔCt between TERT gene and GAPDH gene remained stable.The amplification curve of GAPDH gene in negative standard was normal,while there was no amplification curve of TERT gene.There was a little difference in ΔCt between TERT and GAPDH genes in the positive standard frozen and thawed for 3 and 5 times repeatedly and the positive standard without freezing and thawing,and the CVs of precision in intra-and inter-groups were all less than 1%.Telomerase activity was negative in 19 normal mesenchymal cell samples and positive in 32 breast cancer cell samples,and significant difference in Ct value of TERT gene between them was observed(t=4.236,P <0.001).Conclusion The developed multiplex fluorescence quantitative PCR(Taqman probe) method for the detection of telomerase activity has good stability and precision,which is expected to be used in early diagnosis and gene therapy of tumors.

Objective To study the effects and the therapeutic mechanism of Humulus scandens particles ( HSP ) on the mice with chronic bronchitis ( CB ) . Methods Mice were divided into normal control group, dextromethorphan group or ammonium chloride group, the high-, middle-and low-dose of HSP groups ( These three groups received gavage administration of HSP at 3.00 g??kg-1, 1.50 g??kg-1 and 0.75 g??kg-1,respectively) randomly.The effects of HSP on ammonia-induced cough and trachea phenol red excretion in mice were observed.CB mouse model was established by inhaling smoke and formaldehyde.In total, 60 mice were divided into 6 groups by the method of random digits table: normal control group, model control group, Guilong kechuanning capsule group, the high-, middle-and low-dose HSP groups (n=10 each group).After modeling for 18 days, mice in normal control group and model control group were administered intragastrically with 0. 9% sodium chloride solution. Mice in the high-, middle-and low-dose HSP groups were daily treated with HSP at 3.00, 1.50 and 0.75 g??kg-1 ,respectively, by intragastric administration.Mice in Guilong kechuanning Capsule group were administered with Guilong kechuanning Capsule (600 mg??kg-1), once a day, for 15 days.The tumor necrosis factor alpha ( TNF-α) & interleukin-8 ( IL-8) contents in lung tissues of each group were determined. Results By hematoxylin-eosin (HE) staining, it was found that HSP alleviated the damage of CB in mice.The frequency of cough in 2 min in the normal control group, the high-, middle- and low-dose HSP groups were (17.50±5.38), (11.90±4.46), (12.60±3.47), (9.50±3.24), respectively, and the ecretion of phenol red were (0.52±0.11), (0.65±0.15), (0.64±0.14), (0.67±0.19) μg??mL-1, respectively.The content of TNF-α in lung tissues of the normal control group, model control group,the high dose HSP group was (25.8±6.9), (66.3±11.7), (43.5±7.7) ng??mg-1, respectively.The content of IL-8 in lung tissues of the normal control group, model control group, the high-dose HSP group were (27.1±9.1), (48.2±11.4) and (36.5±8.2) ng??mg-1, respectively.The contents of TNF-α and IL-8 in lung tissues of the model control group were significantly higher than those in normal control group (both P<0.01).Compared with the model control group, high-dose HSP could obviously decrease the contents of TNF-α and IL-8 in lung tissues (P<0.01 or P<0.05). Conclusion HSP can obviously alleviate the pathomorphological changes in mice with CB and has antitussive and expectorant effecs.The therapeutic mechanism of HSP for CB may be related to decreasing the contents of TNF-αand IL-8 in lung tissue.

Elderly individuals make up a considerable large part of the breast cancer population.There are many different biological and pathological characteristics for this population as compared with the younger ones.Evidences on the treatment of elder patients with breast cancer are scarce.This article reviews the studies in the treatment and provides suggestions for the care of elder women with breast cancer.

In this paper, the advances of the treatment for the most common pulmonary mycosis is introduced, and the recommended medications described. It is indicated that amphotericin B is still the first choice for the treatment of severe and rapidly progressive pulmonary mycosis. Lipid formulations of AmB can be used for those refractory or intolerant to the conventional amphotericin B. Fluconazole has proved effective in treating pulmonary candidosis, cryptococosis and coccidioidomysis. Itraconazole has been widely used in the treatment of pulmonary aspergillosis, histoplasmosis ans coccidioidomycosis.

Objective To observe the effect of the reinforcing kidney,nourishing and activating blood on promoting ovulation and improving the endometrium thickness and pregnancy rate in the process of in vitro fertilization-embryo transplantation(IVF-ET).Methods The 63 cases receiving IVF-ET were randomly divided into two groups: the treatment group(n=31) treated with GnRHa/r-FSH/HMG/HCG combined with the herbs of kidney-reinforcing,blood-nourishing and blood-activating in different periods of IVF-ET,and the control group(n=32) with GnRHa/ r-FSH/HMG/HCG alone.The dose of gonadotropin(Gn),endometrium thickness,follicular oocyte number,high-quality oocyte rate,fertilization rate,high-quality embryo rate and pregnancy rate of both groups were observed.Results In the treatment group,Gn dose was significantly lower(P

Adenocarcinoma, Papillary ; complications ; Adult ; Ear Neoplasms ; complications ; Endolymphatic Sac ; pathology ; Facial Paralysis ; etiology ; Humans ; Male

Objective To compare the effects of low-molecular-weight heparin (LMWH,fragmin) and aspirin versus aspirin alone on transient myocardial ischemia and in-hospital prognosis in patients with unstable angina pectoris (UAP).Methods A total of 86 patients with UAP were randomized into a group receiving aspirin (ASP group) a group receiving aspirin plus LMWH (fragmin group) for one week.Results The effective rate was 88.37% in the fragmin group and 67.44% in the aspirin group after one week (P

Objective To study the clinical factors affecting the prognosis of adrenocortical car-cinoma (ACC) in order to guide the diagnosis and treatment of ACC. Methods The clinical data of 40 cases of ACC were retrospectively analyzed. The patients were diagnosed as ACC by histopatholo-gical examination after operation or biopsy, including 19 cases of nonfunctional ACC and 21 cases of functional ACC. The functional ACC included 14 cases of Cushing's syndrome, 4 cases of sexual ab-normality (3 cases of androphany and one case of male sexual precosity) and 3 cases of primary aldo-steronism. The clinical stage included 3 cases in stage Ⅰ , 10 cases in stage Ⅱ , 7 cases in stage Ⅲ, and 21 cases in stage Ⅳ. Eighteen cases underwent complete excision, 10 cases underwent palliative excision, 12 cases underwent expectant treatment (2 cases underwent arterial embolism). All patients were followed up for 2 to 121 months. The total life span and survival rate were calculated, and the dependability between life span and certain indices (tumor size, function, clinical stage and surgical treatment, etc) was compared. The results were statistically analyzed by utilizing the SPSS 11.5 soft-ware. Rank-sum test was used for the two independent samples, one-factor analysis of variance was used for multiple samples and Pearson correlation analysis was used for the correlation of two vari-ances. Results There were no statistical correlations between gender, age, sides and survival time (P>0.05). The survival time of nonfunctional ACC patients was 37.0 months, of which 11 dead ca-ses survived 11.0 months and 8 survival cases survived 73.5 months. The survival time of functional ACC patients was 11.5 months, of which 19 dead cases survived 12.2 months and 2 survival cases survived 4.8 months. The survival time of nonfunctional ACC was significantly longer than that of functional ACC (P<0. 05). The actual survival time was 66 months and 120 months in stage Ⅰ , of which both survived. The actual survival time was (44.9±40. 2) months in stage Ⅱ, of which 4 sur-vival cases survived 59.0 months. The actual survival time was(34. 5±29.8) months in stage Ⅲ, ofwhich 2 survival cases survived 42.0 months. The actual survival time was (7.1±6.7) months instage Ⅳ, of which 2 survival cases survived 8. 0 months. There was significant difference of the sur-vival time among different clinical stages(F=11. 078, P<0. 05). There was no significant difference of survival time between treatment methods[(10.04±0.0) vs (4.5±1.5) months, P>0. 05]. Con-clusions The most important influencing prognostic factors of ACC are function and clinical stage. For ACC in stage Ⅰ and Ⅱ , tumor resection is the most effective treatment, and second surgical oper-ation is recommended for local recurrence. For ACC in stage Ⅲ, radical or extensive surgical operation is recommended, and for ACC in stage Ⅳ, surgical operation has no effect on the prognosis.

Hilar cholangiocarcinoma always presents with a mass, thickness of the bile duct wall or nodules in the bile duct. Computed tomography (CT) can demonstrate the above findings with high partial resolution as well as dilation of the intrahepatic bile duct. CT can show the field and the involvement of adjacent vessels on 3D reconstruction images for hilar cholangiocarcinoma spreading along the lumen of the bile duct. CT images can also demonstrate vessel involvement when evaluating the resectability. When combined with 3D reconstruc-tion, the sensitivity and specificity of CT imaging can reach 90.5% and 91.7% , respectively, for detecting vessel involve-ment.

Mesenchymal stem cells (MSC) is able to differentiate into a variety of epithelial cells,such as alveolar epithelium,tubular epithelium and corneal epithelial cells in vivo and in vitro under the given condition.After transfusion,MSC can effectively repair the injured lung epithelial cells,and relieve inflammation.MSC can also repair the renal tubular cell damage by reducing renal interstitial infiltration of inflammatory cells,promoting regeneration of renal tubular epithelial cells and inhibiting of apoptosis; Biological cornea can be available constructed by using MSC,which can availably repair the injured ocular surface.In the other hands,MSC can express keratinocyte growth factors and other cytokines,promoting limbal stem cell proliferation.In addition,MSC can also differentiate into pancreatic ductal epithelial cells,endometrial epithelial cells,spermatogenic epithelium and human retinal pigment epithelium.MSC has great application potential in epithelial diseases,and provide a good and effective choice in treatment of some epithelial diseases.