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.

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.

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.

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.

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

ObjectiveTo evaluate the efficacy and tolerance of tolterodine in treating detrusor hyperreflexia in patients with spinal cord injury.Methods50 patients of spinal cord injury patients with detrusor hyperreflexia were involved in this study.The data from voiding diary were evaluated 7 days before and after the patients being treated with tolterodine 2 mg twice daily.ResultsAfter 12 weeks of treatment,the functional bladder volume increased from(114.0±44.3) ml to(207.7±54.9) ml,the mean frequency of micturition decreased from(8.9±2.7) /d to(4.6±2.5) /d,the mean volume of incontinence decreased from(646.7±348.9) ml to(426.6±291.3) ml,the mean frequency of incontinence decreased from(7.8±3.1) /d to(4.8±3.2) /d,which were all statistically different before and after treatment.5 patients felt dry mouth,but they could tolerated it.ConclusionTolterodine is an effective and well tolerable antimuscarinic agent for treating detrusor hyperreflexia after spinal cord injury.

OBJECTIVETo explore the molecular biological mechanism of ZHU's Tiaojing Cuyun Recipe (TCR) for treating anovulatory infertility patients with Shen deficiency syndrome (SDS) by observing its clinical efficacy.

METHODSUsing randomized blocking methods, 80 patients were assigned to the treatment group (40 cases) and the control group (40 cases). Patients with regular menstrual cycle started medication from the 5th day of menstruation. Those with irregular menstrual cycle first took progesterone till withdrawal bleeding ,and then started medication from the 5th day of vaginal bleeding. Patients in the treatment group took ZHU's TCR, one dose per day, while those in the control group took Clomifene Citrate (CC), 50 mg per day. Three menstrual cycles consisted of one therapeutic course, a total of 2 courses. Clinical efficacy such as pregnancy rates and abortion rates were observed. Ovulation indices (the maximal diameter of mature follicles, luteinized follicles, ovulational follicles, and the endometrial thickness on the ovulation day), SDS, and integrals of menstrual symptoms were monitored before and after treatment. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) , and estradiol (E2) were determined using chemiluminescent immunoassay before treatment and after on therapeutic course. Serum levels of activin A (ACTA), inhibin B (INHB), and follistatin (FS) were detected using double antibody sandwich ELISA.

RESULTSCompared with the control group, the pregnancy rate was obviously elevated and the abortion rate was obviously lowered in the treatment group (P <0. 05). Ovulation rates of mature follicles and luteinizing follicles decreased more in the treatment group (P < 0.05). Compared with before treatment, integrals for SDS were lower, the maximal diameter of pre-ovulational follicles was increased, and integrals for menstrual symptoms in non-pregnant patients of the two groups were obviously lowered. Meanwhile, the endometrial thickness on the ovulation day was increased in the treatment group after treatment, but reduced in the control group (P < 0.05, P < 0.01). Compared with the control group, integrals for SDS were decreased, and the maximal diameter of pre-ovulational follicles was lowered in the treatment group after treatment (P < 0.05, P < 0.01). Integrals for SDS and the difference in the endometrial thickness on the ovulation day were increased, but the difference in the maximal diameter of pre-ovulational follicles were reduced (P < 0.05, P < 0.01). In the treatment group serum levels of E2 and ACTA increased more after one therapeutic course than before treatment (P < 0.01), but serum levels of INHB and FS decreased more after one therapeutic course than before treatment (P < 0.05). In the control group serum levels of FSH and ACTA increased more, and the serum level of FS decreased more after one therapeutic course than before treatment (P < 0.05, P < 0.01). Compared with the control group, serum levels of FSH and ACTA increased more, and serum levels of INHB decreased more in the treatment group after one therapeutic course than before treatment (P < 0.05, P < 0.01).

CONCLUSIONSZHU'sTCR could improve SDS of anovulatory infertility patients, regulate the follicular development, and elevate the pregnancy rate. Its actions might be associated with regulating their sex hormones, expressions of ovary local factors such as INHB, ACTA, and FS.

Activins ; Clomiphene ; Drugs, Chinese Herbal ; therapeutic use ; Estradiol ; Female ; Follicle Stimulating Hormone ; Follistatin ; Humans ; Infertility, Female ; complications ; therapy ; Inhibins ; Luteinizing Hormone ; Medicine, Chinese Traditional ; Ovarian Diseases ; Ovarian Follicle ; Ovulation ; Progesterone

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.

Objective Investigation of satisfaction of clinical nurses and analysis of the influence factors.Methods The ERG theory of Clayton Alderfer is used as the framework to design the questionnaires,for questionnaire investigation of 249 nurses in the hospital in a stratified sampling,Results The score of general satisfaction of nurses for their work is 3.40±0.58;the satisfaction of nurses for the management of their leaders and teamwork is high,as the satisfaction scores are 4.01±0.76 and 3.60±0.71 respectively;their satisfaction for salary,welfare and social identity is low,as the scores are 2.46±0.86 and 2.50±1.02 respectively.Work satisfaction varies with departments,academic levels,monthly income,work shifts,age,seniority,and objectives for their choice of nursing as profession.Conclusion Managers are prompted to enhance their work in view of the satisfaction scores on survival,mutual relations and growth,as well as factors affecting satisfaction,and take various incentives to improve overall satisfaction of nurses.

AIM: By introducing the thinking and experience of professor Deng Tietao to treat myasthenia gravis, to improve the therapeutic effect and prognosis.benefiting vital qi had been the main line throughout the whole process of phasized that while we grasp spleen-asthenia, we must give consideration to other organs at the same time, regulate and treat the patient with multistrengthening muscle and increasing strength (radix astragali, radix codonopsis pilosulae, rhizoma atractylodis macrocephalae, rhizoma cimicifugae, pericarpium citri reticulatae, radix fici simplicissimae, etc.), the disease had been treated with different dosage of drugs according to the py for one or two years, even though the symptoms had disappeared completely.RESULTS: Because of grasping the root cause, therapy with multi-means and modified prescription according to the varied symptoms, it had a satisfactory effect. An excellent therapeutic effect had been obtained with the decoction for strengthening muscle and increasing strength, which is created by Deng himself.CONCLUSION: Deng's theory holds that myasthenia gravis is mainly due to spleen-asthenia, and related to other four organs (with a predominance of kidney), and the therapy should be strengthening the spleen-qi, so the prescription was selected for strengthening muscle and increasing strength (which is created by Deng himself), with radix astragali being in a lager quantity. Moreover, he paid more attention to psychological therapy and dietary regulation. The Deng's theory is of considerable clinical significance in guiding treatment of myasthenia gravis.