Objective:To study the effect of insulin like growth factor 1 and insulin on the promoter activity of human ?1(Ⅰ) procollagen (COL1A1) gene and the influence of tumor necrosis factor ?(TNF?), interferon ? (IFN?) and interferon ?(IFN?) on the effect Methods:Two deletion constructs,pCOLH0 27 and pCOLH2 5 ,containing portions of 5' flanking region of the gene ( 268～+42 bp and 2 483～+42 bp respectively) ligated to the chloramphenicol acetyltransferase (CAT) reporter gene ,were transiently transfected into human skin fibroblasts The cells were subsequently treated with IGF 1 or insulin, and TNF? or IFN? or IFN? The CAT activity was assessed 24 h after the cytokines added Results:The results showed that 13 nmol/L IGF 1 caused 3 68 fold and 4 04 fold increase in the CAT activity of pCOLH0 27 and pCOLH2 5 respectively and that 2 5 ?mol/L insulin caused 3 69 fold and 3 93 fold increase in the activity of the two plasmids respectively TNF?,IFN? and IFN? reduced IGF 1 or insulin induced promoter activity of pCOLH2 5 Conclusion:IGF 1 and insulin could increase the promoter activity of the human (COL1A1) gene TNF?,IFN? and IFN? could suppress the promoter activity induced by IGF 1 and insulin

Objective To establish the method for detecting human Bocavirus (HBoV) by a real-time fluorescent quantitative PCR (FQ-PCR) and to assay HBoV from nasopharyngeal samples of children with acute respiratory tract infections.Methods The specific primers and Taqman probes of targeted HBoV NP1 gene were designed.The aimed fragment of NP1 gene was amplified with PCR and ligated into a pCEM-T Easy vector to set up the standards.The method for detection of NP1 gene using FQ-PCR was established.The sensitivity and specificity of the method had been tested.Then 195 clinical specimens were subsequently tested.Results The specificity of this method was excellent.The linear amplification of the assay ranges from 10 copies/μl to 10~8 copies/μl.Twelve specimens were tested positive for HBoV in 195 clinical specimens (6.2%).Conclusions The method for detection of HBoV NP1 geneby real-time fluorescent quantitative PCR was established successfully.It can offer fast and reliable diagnosis for HBoV infection.

Objective To analyze the general situation of acupuncture-moxibustion in treating perimenopausal insomnia, as well as the treatment methods and acupoints selection.Method Domestic databases including China National Knowledge Infrastructure (CNKI), Wanfang, VIP, and database of China Biomedical Literature (CBM) were retrieved by using computer, and the literatures related to acupuncture-moxibustion in treating perimenopausal insomnia, published till December of 2014 were recruited. The general data, treatment methods, and acupoints selection of the eligible studies were statistically analyzed.Result Finally 85 studies were enrolled, of which, 76 were clinical trials with acupoints recorded; according to the frequency, the treatment methods were dry acupuncture-moxibustion methods (57.9%), comprehensive treatment of Chinese medicine (23.7%), and integrated method of Chinese and Western medicine (18.4%); the use of body acupuncture ranked the top (51.3%) among the acupuncture-moxibustion methods, followed by auricular point therapy (23.7%), moxibustion (9.2%), abdominal acupuncture (7.9%), and acupoint application (7.9%); the following acupoints ranked the top 10 among the used acupoints: Sanyinjiao (SP 6), Shenmen (HT 7), Baihui (GV 20), Shenshu (BL 23), Taichong (LR 3), Xinshu (BL 15), Taixi (KI 3), Neiguan (PC 6), Pishu (BL 20), and Zusanli (ST 36). The corresponding meridians in order were the Bladder Meridian, Spleen Meridian, Kidney Meridian, Governor Vessel, Liver Meridian, Heart Meridian, Stomach Meridian, Conception Vessel, Pericardium Meridian, Gallbladder Meridian, Large Intestine Meridian, and Triple Energizer Meridian; the used points were mostly distributed on lower limbs, then head and neck, back, upper limbs, and chest and abdomen.Conclusion Currently, the research in this field is focused on clinical prevention and treatment, while the theory and mechanism studies are comparatively less; based on the treatment principle of targeting the fundamental cause, it also presents the principle of selecting points according to meridians and regions in the treatment of perimenopausal insomnia with acupuncture-moxibustion.

The clinical practice in Pediatrics of Chinese medicine is the first step of the students to combine the theory with the clinical practice. The optimization teaching methods for clinical practice in Pediatrics of Chinese medicine was formed by using Delphi method of the extensive consultation from experts in colleges and universities of Pediatrics of Chinese medicine. We used this teaching method in students of Beijing University of Chinese medicine in 2011. Comprehensive evaluation to the implementation of effect was from assessment from students and analysis of exam results. The results showed that based on the survey of the students in the practice, the students in the experimental group give more positive evaluation than the control group on the necessity, satisfaction and harvest in the clinical practice in pediatrics of Chinese medicine.

Objective To evaluate the safety and immunogenicity of group A/C meningococcal polysaccharide conjugate vaccine.Methods The double-blind,randomized,parellel controlled,single central clinical trial was carried out to evaluat safety and immunogenicity of MCV-A/C.Results 4-fold rise rate of antibody to group A,C and A/C were more than 90 percent after MCV.The GMTs of antibody serogroup A and C were more than 1:150 in four trial groups aged 3-5 months,6-23 months,2-15 years and 16-30 years,for which the susceptive subjects seroprotected.There were no significant differences between MCV and the control group in the systemic and local reactions rates.The systemic and local reactions rates after the first,second and third dose of MCV were low.And no severe systemic and local reactions.Conclusion Group A/C MCV was safe and immunogenic for the population≥ 3 months old.Registration National Food drugs Surveillance administrative bureau,Medicine Clinical Experiment Written Directive from a superior" number:2006L04776.

Three compounds were isolated from the extract of Taxus cuspidta Sibe et Zucc with the column chromatography on silica gel and preparative HPLC methods. Their structures were identified according to the physicochemical properties and spectral analysis, and they were identified as (E)-1-methoxy-2-O-(p-coumaroyl)-myo-inositol (1), 2-deacetoxy-7beta, 9a, 10beta-trideacetyltaxinine J (2) and (3aS, 4aR, 6S, 8S, 8aS, 9R, 10R, 10aS)-benz[f]azulene-6, 8, 9, 10 (3H)-terol, 3a, 4, 4a, 5, 6, 7, 8, 8a, 9, 10-decahydro-10a-(1-hydroxyl-1-methylethyl)-1, 8a-dimethyl-5-methylene (3). Among them, compound 1 was a new compound, and compounds 2, 3 were two novel natural products.

OBJECTIVETo observe the impact of acupuncture on the subjective symptom, sleep quality and sleep efficiency in the patients of insomnia differentiated as spleen deficiency or non-spleen deficiency in terms of the spleen and stomach theory.

METHODSSixty patients with insomnia were divided into a spleen deficiency group and a non-spleen deficiency group, 30 cases in each one. In the two groups, acupuncture was applied at Sishencong (EX-HN1), Shenmen (HT 7), Sanyinjiao (SP 6), Zusanli (ST 36) and Tianshu (ST 25). The treatment was given once every other day, five times as one course, and two courses were required. The Athens Insomnia Scale (AIS) was used to observe the changes in the patients' subjective symptoms. The Pittsburgh sleep quality index (PSQI) was adopted to observe the changes of patients' sleep quality. The sleep efficiency was applied to observe the changes in sleep time. The clinical efficacy was determined.

RESULTSThe total effective rate was both 80. 0% (24/30) after 5 and 10 treatments in the spleen deficiency group and was 76. 7% (23/30) and 80. 0% (24/30) respectively in the non-spleen deficiency group. The differences were not significant between the two groups (both P >0. 05). The AIS total scores and PSQI total scores were reduced apparently after 5 and 10 treatments as compared with those before treatment (all P<0. 01). The difference at each time point was not significant between the two groups (all P>0. 05). The sleep efficiency after 5 and 10 treatments was all improved as compared with that before the treatment in the two groups (all P<0. 01).

CONCLUSIONAcupuncture based on the spleen and stomach theory achieves possibly the same clinical efficacy in the patients of insomnia differentiated as those with spleen deficiency and non-spleen deficiency. It relieves the subjective symptoms and improves the sleep quality and sleep efficiency in the patients.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Sleep ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Spleen ; physiopathology ; Stomach ; physiopathology ; Treatment Outcome

Objective To identify the glycometabolism features ofC57BL/Ksj db/+ mice at gestational period and provide the evidence for the mice used as the animal model of gestational diabetes mellitus (GDM).Methods Ten C57BL/Ksj db/+ female mice and ten wild female mice were randomly selected and mated with wild male mice respectively.Day 3 before mating,days 6,11 and 17 of pregnancy,and day 6 after delivery represented pre pregnancy,early-pregnancy,mid-pregnancy,late-pregnancy and post-pregnancy periods,respectively.The levels of glucose tolerance and fasting insulin of female mice were compared at progestation,different gestational periods and puerperium between the two groups.Days 6,11,14,17 and 20 (before labor) of pregnancy served as five time-points of pregnancy,leptin levels of female mice of the two groups were dynamically monitored during pregnancy,and analyzed at different gestational ages.The data were analyzed by Tukey test of one-way ANOVA.Results (1) Compared with the normal pregnant mice,the levels of glucose tolerance of C57BL/Ksj db/+ female mice were significantly decreased,while the levels of fasting insulin were dramatically increased with the progression of pregnancy [day 6 of pregnancy,(8.6 ± 0.6) mU/L vs (7.5 ±1.1) mU/L; day 11 of pregnancy,(9.8±0.6) mU/L vs (8.5± 1.1) mU/L; day 17 of pregnancy,(10.4±0.8) mU/L vs (9.9±0.5) mU/L; t=1.859,4.056 and 3.078,P=0.085,0.000 and 0.000] and recovered to the pre-pregnancy level after delivery.(2) The level of leptin in serum of C57BL/Ksj db/+ pregnant mice was significantly higher than that at non-pregnant period [pre-pregnancy and days 6,11,14,17 and 20 of pregnancy:(911.0±238.8) ng/L,(1 342.2± 132.3) ng/L,(1 821.9±238.2) ng/L,(1 816.3± 142.3) ng/L,(1 752.4± 126.6) ng/L,and (1 926.4±61.6) ng/L,respectively; t=3.887,8.210,8.161,7.585 and 9.153,P=0.005,0.000,0.000,0.000 and 0.000],but showed no apparent change with the gestational ages after mid-gestation.Conclusion Changes of glycometabolism and the levels of leptin and insulin of C57BL/Ksj db/+ pregnant mice during pregnancy are similar to those of GDM patients,the levels of fasting insulin are dramatically increased with the progression of pregnancy,and return to normal after delivery,the level of leptin is higher than that at non-pregnant period,but is not correlated with the gestational ages.C57BL/Ksj db/+ pregnant mice can be used as the animal model of GDM.

Objective To detect the double DNA of trisomy 21 syndrome patients by denaturing high-performance liquid chro-matography(DHPLC) in order to rapidly diagnose trisomy 21 syndrome .Methods To amplify DNA fragments of three short tan-dem repeats of D21S111 ,D21S1411 and D21S1412 using corresponding primers .Then DHPLC was introduced to analyze the DNA fragments in the temperature of 50 ℃ .Results DHPLC elution profiles of D21S111 ,D21S1411 and D21S1412 of normal control showed one peak or two peaks with the same altitude .However DHPLC elution profiles of 21 trisomy syndrome patients showed two peaks of different altitudes ,which one′s altitude was twice than another .Conclusion DHPLC is a sensitive ,convenient ,auto-matic and highly-efficient method to diagnose trisomy 21 syndrome and can be widely used in the clinic diagnosis .

Objective:A study was conducted to investigate the effect of Prunella vulgaris sulfated polysaccharide (PVSP) on the expression of angiogenic growth factors (bFGF, VEGF, and IL-8) and angiogenesis in hepatocellular carcinoma. Methods:ELISA as-say was used to observe the effects of PVSP and the negative control drug Lentinan (LNT, non-sulfate radical drug) on secretions of the angiogenic growth factors, namely, bFGF, VEGF, and IL-8, in HepG2 cells in vitro. In an in vivo experiment, the microvessel density in hepatocellular carcinoma tissue sections treated with PVSP and LNT was calculated, analyzed, and compared with the microvessel den-sity in the phosphate-buffered saline (PBS) control. Results:Compared with the PBS control group, PVSP at 200μg/mL inhibited bF-GF secretion (P<0.01), whereas LNT failed to affect bFGF secretion. Neither PVSP nor LNT affected the secretions of VEGF and IL-8. In vivo results showed that PVSP at 200 mg/kg reduced the microvessel density in tumor tissue sections (P=0.03), whereas LNT at 10 mg/kg failed to affect microvessel density. Conclusion:Inhibition of bFGF secretion is a probable mechanism underlying the preven-tive effect of PVSP on hepatocellular carcinoma angiogenesis.