Objective: To establish a method for determination of forsythin in Shuanghuanglian Oral (Flos Lonicerae, Radix Scutellariae, Fructus Forsythiae, etc.) Methods: After processing by RP SPE (reverse phase solid phase extraction), the sample solution was measured by RP HPLC. The chromatographic conditions were: Prodigy ODS (3) (150?4.6mm,5?m) column as analytic column; MeOH H 2O HAc(40∶60∶1,V/V) as mobile phase; detection wavelength at 227nm; column temperature at 35?C ; flow rate at 1.0mL?min -1 . Results: The linear range of forsythin was 0.1～2.0?g, r =0.9997. The average recovery was 102.2%, RSD =0.61%( n =6). Conclusion: The method is simple, accurate, reproducible and can be used to enhance the quality control of this drug.

Objective: To establish the method for determination of taurocholic acid in the Traditional Chinese Medicine Shedanchuanbei Oral Liquid and snake bile. Methods: The sample was prepared as mixed solution containing methanol and KH 2PO 4. The mixed solution was injected into Sep Pak C 18 cartridge for the purpose of sample purity. In this processing, the substances which having strong retain action and could harm analytic column were hold in the Sep Pak C 18 cartridge. The eluting solution that the Sep Pak C 18 cartridge had be over loading for taurocholic acid was used as the test solution. The test solution was measured by RP HPLC. The chromatographic conditions were as followed: Supelcosil LC 8 column(150mm?4.6nm,5?m) as analytic column, detect wavelength at 203nm, and MeOH 0.4%KH 2PO 4 mixed solution(56∶44, V/V ) as mobile phase. The inject volume was 50?L. Results: The linear response range of sodium taurocholate was from 0.0253mg?mL -1 to 0.253mg?mL -1 , and the correlation coefficient was 0.9999. The average recovery rate was 101.3%, RSD was 0.40%( n =6). Conclusion: This method was simple, efficient and suitable to the quality control for Shedanchuanbei Oral Liquid and snake bile.

Objective To established a method for the determination of puerarin. Method The sample solutions were pretreated by solid phase extraction (SPE): the specimens were eluted and the cartridges saturated gradually by them in the SPE, but the strongly absorbed impurities were held by the cartridges all the same, and then the effluents were measured by HPLC. Results The linear range of puerarin was 0.205～ 2.464? g, r=0.9999. The average recovery of puerarin in Xintong Oral Liquid, Radix Purerariae and Ganmao Zhike Capsule were 99.5 % , 99.9 % and 98.5 % , RSD were 1.67 % , 1.45 % and 0.95 % (n=5) respectively. Conclusion The method is simple and accurate and can be used for the determination of puerarin in the Chinese patent medicines containing Radix Purerariae. 

Objective To investigate the potential of AFP-targeted ultrasmall superparamagnetic particles of iron oxide (USPIO) molecular probe in specific detection of hepatocellular carcinoma (HCC) with MRI.Methods The targeted probe was synthesized by conjugating AFP antibody with modified USPIO.Two groups treated with AFP-USPIO and USPIO were set up in the study.The HepG2 cells were incubated with AFP-USPIO or USPIO (100 μg/ml) respectively with the dosage of 50 μ1,100 μl or 150 μl for 4 hours,followed by MR imaging in vitro.The signal-noise ratio (SNR) of the cells on T2-weighted image (T2WI) was measured.The rat models with orthotopic HCC were divided into two groups with 5 rats for each group at random.Pre-and post-contrast enhanced (after 1 hour) MR imaging were performed with caudal vein injection at a dosage of 20μg/ml.The contrast noise ratio (CNR) on T2WI and the difference of CNR between pre-and post-enhancement or between both groups were calculated.The relationship of SNR or CNR with the iron particles in cells or tumors was confirmed by Prussian blue iron staining.Results Cytology experiment showed the SNR in both groups was decreased with the increase of the dosage of AFP-USPIO or USPIO,indicating statistically significantdifference in SNR among three different doseage groups (P＜0.05).Prussian blue iron staining showed that the iron particles in cells were increased with the increase of AFP-USPIO dosage,and was negatively correlated with SNR (P=0.00,r=-0.926).However,the iron particles were less in cells in USPIO group.The CNRs of liver tumors in Wistar rat of pre-and post-AFP-USPIO injection were 2.05±0.88 and 0.96±0.31 respectively,indicating a significant difference (P=0.028,t=3.380).However,the CNRs in USPIO group,2.25±1.50 and 2.57±1.49,showed no statistical difference (P=0.275,t=1.263).The CNR after enhancement also had a statistical difference between both groups(P=0.042,t=3.487).Pathological results confirmed more iron particles in tumor tissues in AFP-USPIO group,whereas less in USPIO group.Conclusion AFP-USPIO molecular probes can initiatively target to the HepG2 cells and the liver cancer of rat models expressing AFP,which may help to achieve the specificity of MR imaging in the diagnosis of HCC.

Objective To investigate the relative risk factors for early abortion among singleton pregnancies after assisted reproductive technology (ART) treatment. Methods A retrospective analysis was performed on 1636 singleton pregnancies, including 196 early abortion cases and 1195pregnancies with live birth after exclusion of those lost cases during follow-up, or complicated with uterine deformity, or oocyte receptor, or late abortion, or incomplete medical record, following in vitro fertilization(IVF)/ intracytoplasmic sperm injection (ICSI) treatment and the risk factors of early abortion were investigated. The early abortion rate was also compared between fresh IVF/ICSI group and frozen embryo transfer (FET) group (n=386). Results Multivariate Logistic regression analysis indicated that elder women (OR= 1. 143,95%CI: 1. 096-1. 196) and patients with polycystic ovarian syndrome (OR = 4. 309,95 % CI : 2. 564-7.243) were risk factors of spontaneous early abortion,and high mean score of transferred embryos (MSTE) (OR = 0. 808, 95% CI: 0. 717-0. 912) and endometrial triple-lined pattern on the day of human chorionic gonadotropin (hCG) administration (OR=0. 431, 95% CI: 0. 243-0.764)were protective factors. Significant difference were found in the maternal age [(32.22±4. 10) yrs vs (30.28±3. 66) yrs],the duration of infertility [(5. 90±4.26) yrs vs (5.20 ± 3. 32) yrs], basal serum follicle-stimulating hormone (FSH) level [(6. 35 ±2.30) mIU/ml vs (5.95±2.12) mIU/ml], number of transferred embryos (2. 31±0. 51) vs (2. 18±0.49), serum estradiol level on the day of hCG administration [(2467. 1 ± 1588. 8) pg/ml vs (2934. 5 ±1785.2) pg/ml] and MSTE (7.03 ±1.35 vs 7.74 ± 1.25) between the abortion group and livebirth group (all P＜0. 05). The spontaneous abortion rate was higher in the FET group than in the fresh embryo transfer group [17. 36%(67/386) vs 13.02% (213/1636), χ2 =4. 296, P=0. 023].Conclusions Women at elder age, or with long duration of infertility, high basal FSH level,polycystic ovarian syndrome, low MSTE, non-triple-lined pattern of endometrium on the day of hCG administration are at risk of spontaneous early abortion in pregnancies after ART. The rate of spontaneous abortion is higher in FET group than in fresh IVF/ICSI group.

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer, the physical status of HPV-16 in patients with cervical lesions, and the role of HPV-16 integration in cervical carcinogenesis were investigated. HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women. Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection, including 112 specimens with cervical cancer, 151 specimens with CIN I, 246 specimens with CIN and 120 specimens with CINIII. The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection. The frequency of HPV-16 pure integration was 65.18% (73/112), 56.57% (47/120), 23.58% (58/246) and 7.95% (12/151) in cervical cancer, CINIII, CINII and CINI patients respectively. In situ hybridization was performed on some paraffin-embedded sections of CINII, CINIII and cervical cancer to verify the physical status of HPV-16 infection. Significant difference was observed between cervical cancer and CIN I, CINII, CINIII in the frequency of HPV-16 integration (P<0.01). It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer. In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration. The application of testing HPV genotyping and physical status based on detection of HC-II HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer,the physical status of HPV-16 in patients with cervical lesions,and the role of HPV-16 integration in cervical carcinogenesis were investigated.HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women.Multiple PCR was done to detect physical status of HPV-1 6 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection,including 112 specimens with cervical cancer,151 specimens with CIN Ⅰ,246 specimens with CIN Ⅱ and 120 specimens with CINⅢ.The results showed that there were 112 cervical cancer samples (56.57％ of total cervical cancer patients) with HPV-16 infection.The frequency of HPV-16 pure integration was 65.18％ (73/112),56.57％ (47/120),23.58％ (58/246) and 7.95％ (12/151) in cervical cancer,CINⅢ,CIN Ⅱ and CIN Ⅰ patients respectively.In situ hybridization was performed on some paraffin-embedded sections of CIN Ⅱ,CINⅢ and cervical cancer to verify the physical status of HPV-16 infection.Significant difference was observed between cervical cancer and CIN Ⅰ,CIN Ⅱ,CINⅢ in the frequency of HPV-16 integration (P＜0.01).It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer.In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration.The application of testing HPV genotyping and physical status based on detection ofHC- Ⅱ HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.

Huperzine A (Hup-A) is a poorly water-soluble drug with low oral bioavailability. A self-microemulsifying drug delivery system (SMEDDS) was used to enhance the oral bioavailability and lymphatic uptake and transport of Hup-A. A single-pass intestinal perfusion (SPIP) technique and a chylomicron flow-blocking approach were used to study its intestinal absorption, mesenteric lymph node distribution and intestinal lymphatic uptake. The value of the area under the plasma concentration-time curve (AUC) of Hup-A SMEDDS was significantly higher than that of a Hup-A suspension (<0.01). The absorption rate constant () and the apparent permeability coefficient () for Hup-A in different parts of the intestine suggested a passive transport mechanism, and the values ofandof Hup-A SMEDDS in the ileum were much higher than those in other intestinal segments. The determination of Hup-A concentration in mesenteric lymph nodes can be used to explain the intestinal lymphatic absorption of Hup-A SMEDDS. For Hup-A SMEDDS, the values of AUC and maximum plasma concentration () of the blocking model were significantly lower than those of the control model (<0.05). The proportion of lymphatic transport of Hup-A SMEDDS and Hup-A suspension were about 40% and 5%, respectively, suggesting that SMEDDS can significantly improve the intestinal lymphatic uptake and transport of Hup-A.