OBJECTIVE:To establish a method for simultaneous determination of geniposide,berberine hydrochloride and ba-icalin in Annao pill. METHODS:HPLC was performed on the column of Inertsil ODS-3 C18 with mobile phase of acetonitrile-0.2% phosphoric acid solution (gradient elution) at a flow rate of 1.0 ml/min,the column temperature was 30 ℃,the detection wavelength was 230 nm,and the injection volume was 10 μl. RESULTS:The linear range was 0. 041 0-0.820 0 μg for geniposide (r=0.999 9),0.037 6-0.752 0μg for berberine hydrochloride(r=0.999 9)and 0.095 2-1.904 0μg for baicalin(r=0.999 9),respec-tively;recoveries were 97.31%-100.41%(RSD=1.10%,n=6),98.32%-101.76%(RSD=1.20%,n=6) and 99.16%-101.30%(RSD=0.76%,n=6),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the quality control of Annao pill.

Objective: To establish a method for the simultaneous determination of paeoniflorin, hesperidine, baicalin and chry-sophanol in Xiaoshi Lidan capsules by dual wavelength HPLC. Methods:An Inertsil ODS-3 C18 column (150 mm × 4. 6 mm,5 μm) was used. The mobile phase was acetonitrile-0. 2% phosphoric acid with gradient elution. The flow rate was 1. 0 ml·min-1 , the de-tection wavelength was 230nm for paeoniflorin, hesperidine and baicalin, and 280nm for hrysophanol, and the column temperature was 40℃. Results:The linear range of paeoniflorin, hesperidine,baicalin and chrysophanol was 0. 030-0. 592 μg(r=0. 999 9), 0. 063-1. 264 μg(r=1. 000 0), 0. 205-4. 094 μg(r=1. 000 0) and 0. 008 2-0. 164 μg(r=1. 000 0), respectively. The average recovery of the four components was 99.49% (RSD =0.85%), 99.74%(RSD =0.71%), 99.75%(RSD =0.47%) and 99.08%(RSD =1. 28%), respectively. Conclusion:The method is simple and accurate, which can be used for the content determination of paeoniflor-in,hesperidine,baicalin and chrysophanol in Xiaoshi Lidan capsules.

Objective To reconstitute a transactivation system in yeast (yeast model) for screening the pesticides acting on ecdysone metabolism route and eventually influencing the process of ecdysis. Methods The fragment of 5 times repeated EcRE from Drosophila melanogaster was synthesized and the HSP27 promoter from D. melanogaster genome was amplified with PCR. The two sequences were connected and followed by a reporting gene——green fluorescence protein(GFP) gene. The EcRE-HSP27 promoter-GFP fragment was inserted into the expression plasmid pPIC3.5 and integrated into the yeast chromosome to construct yeast A. EcR and USP coding sequences of Aedes albopictus were synthesized, and these two fragments were inserted into Pichia pastoris expression plasmid pGAPZ as two respective reading frames. The two reading frames were integrated into Pichia pastoris chromosome in another recombinant site(pGAPZ and pPIC3.5k share different recombinant sites while being integrated into Pichia pastoris yeast chromosome). EcR and USP were constituted and expressed in the yeast. This recombinant yeast was called yeast B. The model yeast was thus constructed. A known ecdysone agonist-tebufenozide was used to test the yeast model. The effect of tebufenozide on the model yeast was observed under fluorescent microscope. Semi-quantitative RT-PCR was used to test the transcrip-tion level of GFP in the tebufenozide affected yeast and the control. Results In the model yeast, the intracellular expressed EcR and USP constituted EcR/USP heterodimer interacting with EcRE, the expression of GFP was activated, and green fluorescence was observed in model yeast under fluorescent microscope. Tebufenozide affected model yeast showed less fluorescence in comparison to the control model yeast, indicating that the transcription of GFP was suppressed by tubufenozide. Yeast housekeeping gene Actin-1 was used as inner control, semi-quantitative RT-PCR was operated and the result was scanned. The ratio of the brightness of GFP to Actin-1 was calculated automatically, and that of tubufenozide added yeast and the control yeast was 0.614 and 1.134 respectively. This result showed a low transcription level of GFP in tebufenozide affected model yeast, comparing to that of the control. Conclusion The ecdysone-related transacting system in yeast has been constructed, and the model yeast can be used to screen the ecdysone agonists which can act on the ecdysone metabolic route.

AIM: To establish the method of fingerprinting analysis on Zedoary turmeric oil by GC.Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them.Based on the cluster analysis,all the Zedoary turmeric oils could be divided into two categories,self-made and market purchasing oil,the latter oils purchased from Liaoning province and Jiangsu province varied considerably.Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the market purchasing Zedoary turmeric oil,quality control requires serious consideration.

AIM: To establish the method of fingerprinting analysis onZedoary turmeric oil by GC. Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them. Based on the cluster analysis, all the Zedoary turmeric oils could be divided into two categories, self-made and market purchasing oil, the latter oils purchased from Liaoning province and Jiangsu province varied considerably. Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the marketpurchasing Zedoary turmeric oil, quality control requires serious consideration.

Objective To discuss the effective factors and metastasis rules of cervical lymph node of papillary thyroid microcarcinoma,aimed to provide references for indications and range of lymph node dissection.Methods Reviewed the clinical information and pathological material of primary treatment of 1 180 cases of papillary thyroid microcarcinoma patients in China-Japan Union Hospital of Jilin University from June 2008 to December 2011 by retrospective analysis,futher summared cervical lymph node metastasis rules and effective factors.Results The lymph node metastasis rate of 1 180 patients with papillary thyroid microcarcinoma was 29.6％,which of these of the central and lateral region were 26.4％ and 15.2％.Central region lymph nodes metastasis rates in male and female were respectively 44.9％,21.8％ (P ＜ 0.05),and were 39.8％,9.0％ in lateral region (P ＜ 0,01).Lymph node metastasis rates of patients whose age ≤45years old and ＞45 years old were 30.3％,17.9％ (P ＜0.05).The lymph mode metastasis rate which diameter of tumour(D) ≤0.5 cm or 0.5 cm ＜ D ≤ 1 cm in central region were 20.2％,33.2％ (P ＜ 0.05).The lymph node metastasis rate of single and more nidi respectively were 15.7％,45.0％ in central region.The lymph node metastasis rate of tumors invasion or not were 44.4％ and 14.2％ in central region (P ＜0.05).The metastasis rates were 45.5％ and 4.3％ in the lateral region when the lymph node infringe or not in central region.Conclusions Lymph node metastasis happens relatively more common in papillary thyroid microcarcinoma patients with the factors such as male,young age,multiple nidi,long diameter of tumor and invasion of tumor envelope.So when doctors establish the indications and range of lymph node dissection,it's need to pay attention to the high risk factors seriously which effect lymph node metastasis of papillary thyroid carcinoma,and establish the lymph node dissection strategy individually.

Objective To approach the guide of ultrasound-guided minimally invasive biopsy technique in diagnosis and treatment of thyroid diseases. Methods One hundred and seventeen cases (a total of 131 nodules, nodules 0. 5-1.5 cm in diameter, with an average diameter of approximately 0. 8 cm) with thyroid nodules , under Ultrasound-guided minimally invasive biopsy techniques from October 2008 to July 2009 were analyzed. Result One hundred and seventeen cases of patients with biopsy derived satisfaction were not ap-parent discomfort and complications after biopsy. Across 59 cases of surgical treatment in patients with biop-sy, and preoperative biopsy and postoperative pathologic is not fully consistent in only two cases, 58 patients without surgical treatment, who is combined with symptoms, palpation, serum thyroxine and color Doppler ultrasound and other laboratory examinations, and regularly review treated with symptomatic treatment, the symptoms, and color Doppler ultrasound imaging were much better under the treatment. 8 cases of patients in 117 cases of minimally invasive biopsy have cervical lymph node biopsy at the same time, biopsy patholo-gy and postoperative results are consistent with postoperative pathologic (7 cases of metastatic carcinoma, 1 case of reactive hyperplasia), accuracy was 100%. Conclusion Uhrasound-guided minimally invasive bi-opsy technique have important guiding for diagnosis and treatment of thyroid diseases, consistent with the high rate of clinical, especially for the preoperative diagnosis of small nodules, to avoid unnecessary over-treatment, can also reduce the misdiagnosis of thyroid cancer. While this technology is simple, safe and ac-curate, which have clinical value in the thyroid surgery.

Objective To find new way to reduce non-recurrent laryngeal nerve (NRLN) injuries by applying intraoperative neuromonitoring(IONM) to identify NRLN in thyroidectomy. Methods Records of 279 patients who underwent complex thyroidectomy by applying IONM to identify and monitor RLN from Mar. 2009 to Jan. 2010 were veviewed. We proposed the skills to identify and monitor NRLN and predict RLN varition through exploring vagus nerve and RLN before RLN dissection. Results 6 cases NRLN located on the right side were all accurately identified by IONM, thus no injury of NRLN occurred during thyroid operations. Conclusions NRLN is difficult to be predicted preoperatively and identified by naked eyes. The application of IONM to predict, identify and monitor NRLN could remarkably reduce the possibility of NRLN injury.

ObjectiveTo investigate the mechanism and prevention of invisible injury of recurrent laryngeal nerve (RLN) system in thyroid surgery, with the application of intraoperative neuromonitoring ( IONM ) system. MethodsThe type of invisible RLN injury and its protection with the application of IONM system were analyzed. ResultsThe causes of invisible RLN injury mainly included stretching of Berry ligament or the tumor,contusion, thermal injury, cutting of silk and suction injury. RLN invisible injury was recoverable through neurotrophic and symptomatic treatment. No permanent vocal cord paralysis occurred. ConclusionsWith the application of IONM system, some invisible type of RLN injuries can be found. The risk of RLN injury can be reduced if the surgical techniques are improved with the development of study on mechanism of IONM system.

Objective To analyze prevention and management of lymphatic fistula after cervical lymph node dissection in thyroid carcinoma.Methods Clinical data of 9 cases of lymphatic fistula following neck dissection were analyzed retrospectively from Jan.2004 to Apr.2009.Results Of the 9 cases,4 cases had chylous fistula(3 were on the leftside and 1 was on the rightside),1 case had chylotborax,1 case had pleural effusion lymph,and 3 cases showed light yellow lymph.AII patients were cured finally by conservative methods.Conclusions Most lymphatic fistula can be cured by non-surgical treatment.Surgeons need to be familiar with the anatomic structure of neck lymphatic vessels.