Objective Based on real-time PCR technique and ring primers,to establish a simple,accurate,cost-effective and easily standardized quantitative assay for quantification of HER2 mRNA,and apply to provide medication guidance for clinical tumor personalized molecular targeted therapy.Methods Screening reference gene which was stable expression in breast cancer,and optimizing the PCR reaction system.Then a real-time PCR with Eva Green for quantification of the mRNA expression levels of HER2 gene was developed.The specificity,sensitivity and reproducibility of the method were evaluated 87 specimens including 55 liquid nitrogen-frozen breast cancer tissues and 32 normal tissues were detected by the real-time quantitative reverse transcription (FQ RT)-PCR and immunohistochemistry(IHC).Results The standard curve of the method indicated a good linear relationship between the Ct value and the template concentration with the correlation coefficient being 0.997.The linear range of the system was from 101 to 106 copies/μl and the lower detection limit was 101 copies/μl.It had a high sensitivity and good specificity.The inter-assay coefficients of variation of HER and RPL37A genes were (5.93 ± 0.57)％ and (5.11 ± 0.59)％,(2.49 ±0.81)％ and (2.98 ±0.97)％ respectively.The intra-assay coefficients of variation were (5.76 ±0.58)％and (7.71 ±0.61)％,(3.75 ±0.76)％ and (4.40 ±0.96)％ respectively.Using the optimized FQ RTPCR system,HER2 gene of 87 specimens was quantificated.The sensitivity of the assay was 96.36％ (53/55),the specificity was 78.13％ (25/32),the positive predictive value was 88.33％ (53/60),the negative predictive value was 92.59％ (25/27),and the total coincidence rate between FQ RT-PCR and IHC was 89.66％ (78/87).The correlation of the results between the FQ RT-PCR and IHC was good (Kappa =0.770,P ＞ 0.05).Conclusions The method can quantify the mRNA expression levels of HER2 gene rapidly and cost-effictively with high sensitivity and specificity.It can be applied to clinical molecular diagnosis with attractive prospect.

OBJECTIVETo observe the effects and safety of clinical common symptoms of active Crohn's disease at the mild and moderate stages treated with acupuncture and moxibustion.

METHODSOne hundred and two patients were randomly divided into an observation group and a control group, 51 cases in each one. Herb-partition moxibustion and acupuncture were used in the observation group. Herbal cakes with(),(),(),(),(),() and() as the main ingredients were used on Tianshu (ST 25), Qihai (CV 6) and Zhongwan (CV 12); acupuncture was used at Zusanli (ST 36), Shangjuxu (ST 37), Sanyinjiao (SP 6), Taixi (KI 3), Gongsun (SP 4) and Taichong (LR 3). Bran-partition moxibustion and shallow acupuncture were applied in the control group. Wheat bran-partition moxibustion in the shape of cake was implemented on the same acupoints as the observation group; shallow acupuncture was used 1~2 cm next to the acupoints. Treatment was given three times a week for 12 weeks, 36 times totally. Clinical effects of eight symptoms were evaluated by TCM symptom score, and the symptoms included abdominal pain (degree, frequency, time), diarrhoea (number, shape and frequency), fatigue, anorexia, borborygmus andflatus, fear of cold, soreness and weakness of waist and knees, tenesmus.

RESULTSAfter 12-week treatment, the eight symptoms were improved apparently (all<0.05), with the scores of abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia in the observation group decreasing more obviously (all<0.01). There was no statistical significance about the other symptom scores between the two groups (all>0.05).

CONCLUSIONSHerb-partition moxibustion combined with acupuncture achieve safety and efficacy for mild and moderate Crohn's disease, and can improve the clinical common symptoms. Furthermore, their effects on abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia are better than those of bran-partition moxibustion and shallow acupuncture.