Evaluation of embryo toxicity of Shuanghuanglian based on human placental barrier model / 中国药理学与毒理学杂志

OBJECTIVE To evaluate the embryo toxicity of Shuanghuanglian (SHL) by the combination of a human placental barrier model and embryonic stem (ES) cell test model.METHODS A human placental barrier model was set up by placenta slice culture and Ussing chamber.SHL 0.2,0.4,0.8,1.6,3.2,6.4 and 12.8 g· L-1 was added into the maternal side of the human placental model,respectively.All the media was collected respectively from the matemal side and fetal side 60 min later and taken as the SHL containing medium.ES cells (D3 line) and embryonic fibroblast cells (BALB/c 3T3) were cultured with the SHL containing medium respectively from the maternal side and the fetal side for 10 d.Cell viability was detected by MTT assay,and 50％ survival inhibitory ratio of ES and 3T3 cells by SHL was calculated.ES cells were incubated with the SHL containing medium from the matemal side or fetal side when they differentiated to cardiac myoblasts using hanging drop-suspension-attachment method.Messenger RNA of myosin heavy chain genes (β-MHC) was detected by Q-PCR for differentiation ratio,and 50％ differentiation inhibitory ratio of ES cells by SHL was calculated.A statistics formula was used for prediction of SHL embryotoxicity potential.RESULTS The IC50 of SHL in the matemal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 1.97,0.84 and 0.48 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,SHL had weak embryo toxicity.However,the IC50 of SHL in the fetal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 3.19,2.57 and 0.95 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,the supernatant containing SHL that went through the placental barrier had no embryo toxicity.CONCLUSION SHL is safe in the test concentration range during pregnancy.It is more scientific to evaluate embryo toxicity of drugs by ES cell test with the samples obtained through the placental barrier during pregnancy.

Simultaneous Determination of the Contents of 8 Ingredients of Shuanghuanglian Freeze-dried Powder in Pla-cental Perfusate / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of the contents of Shuanghuanglian freeze-dried powder in placental perfusate. METHODS:HPLC was performed on the column of Agilent Zorbax-C18 with mobile phase of acetonitrile-1%formic acid aqueous solution(gradient elution)at flow rate of 1.0 ml/min,the internal standard was puera-rin,detection wavelength was 280 nm,column temperature was 25 ℃ and sample volume was 10 μl. RESULTS:There was a lin-ear range between linear ranges and peak area of 8 ingredients(r≥0.999 0);RSDs of within-day and inter-day precision tests were no more than 1.9%,repeatability tests was no more than 7.3%;average recoveries were in the range of 92.73%-112.37%(RSD=3.2%-8.2%,n=6);and average matrix effects were 90.33%-105.78%(RSD=3.2%-8.0%,n=6). CONCLUSIONS:The method is rapid,sensitive and specific,and can be used to the simultaneous determination of the contents of 8 ingredients of Shuan-ghuanglian freeze-dried powder in placental perfusate.

Invasion and Metastasis Ability of Endometrial Stromal Cells in Endometriosis / 天津医药

Objective To analyze the invasion and metastasis of endometrial stromal cells in endometriosis through assessing the transcription levels of COX-2, PGE2, Snail and E-cadherin mRNA. Methods Ectopic and eutopic endometri?al tissues from patients with EMs (endometriosis) or uterine fibroids were collected and cultured. The fourth generation of pu?rified endometrial stromal cells were used for research, and the transcription levels of COX-2, PGE2, Snail and E-cadherin mRNA were analyzed by RT-PCR. Results There were no statistical differences in transcription levels of COX-2, PGE2, Snail and E-cadherin mRNA in eutopic endometrial stromal cells from patients with EMs or uterine fibroids between secra?tion stage and proliferation stage(P>0.05). The transcription levels of COX-2, PGE2 and Snail mRNA were significantly in?creased while transcription level of E-cadherin mRNA was decreased in eutopic and ectopic endometrial stromal cells in pa?tients with EM compared with those in patients with uterine fibroid. And the difference was statistically significant(P<0.05 or P<0.01). The transcription levels of COX-2, PGE2 and Snail mRNA were obviously higher while transcription level of E-cadherin mRNA was lower in ectopic endometrial stromal cells than those in eutopic endometrial stromal cells from pa?tients with EMs. And the difference was statistically significant(P<0.05 or P<0.01). Conclusion The invasion and me?tastasis of endometrial stromal cells of patients with EMs were remarkable than those cell of patients without EMs. What ’s more, the invasion and metastasis in ectopic endometrial stromal cells were more obvious than those cells in the pathogenesis of EMs.

Study on ingredients of Scutellaria Radix extract penetrable through placental barrier of pregnant rat / 中国中药杂志

<p><b>OBJECTIVE</b>To study ingredients penetrable through placental barrier by administering pregnant rats with Scutellaria Radix extract using HPLC-MS.</p><p><b>METHOD</b>Rats in early, middle and late pregnancy were intragastrically administered with Scutellaria Radix extract for 5 days. Maternal plasma and embryonic tissues were obtained at 1.5, 12 h after the final administration of Scutellaria Radix extract to determine ingredients of biological specimens.</p><p><b>RESULT</b>Under the optimum experimental conditions, seven compounds were detected in all pregnant rat plasma, specifically including baicalin, wogonoside, baicalein, wogonin and oroxylin A. The seven compounds were also discovered in embryonic tissues of rats in early pregnancy, including the five detected compounds. But they were not detected in embryonic tissues of rats in middle pregnancy, and six compounds except baicalein were detected embryonic tissues of rats in late pregnancy.</p><p><b>CONCLUSION</b>Ingredients contained in Scutellaria Radix are detected in pregnant rats at different stages, except those in middle pregnancy, indicating a potential in utero exposure in case of oral administration of Scutellaria Radix pregnancy. Therefore, a study of embryotoxicity shall be continued to evaluate the safety of Scutellaria Radix extract.</p>

Ingredients of Shuanghuanglian injection powder permeation through placental barrier of rat in pregnancy / 中国中药杂志

<p><b>OBJECTIVE</b>Use HPLC to study the permeation of ingredients of Shuanghuanglian injection powder (SHL) through placental barriers of rats at different stages of pregnancy.</p><p><b>METHOD</b>The pregnant rats were administered SHL for 5 d through caudalis vena at different stages of pregnancy. Plasma and embryonic tissues were obtained 12 h after the final administration of SHL. The componds in biological specimen were identified by HPLC.</p><p><b>RESULT</b>Baicalin, luteolin and wogonoside were the main compounds in plasma. Wogonoside retained in first trimester embryonic tissues, and baicalin retained in the embryonic tissues of different pregnant stages.</p><p><b>CONCLUSION</b>Baicalin is the main compound of SHL through placental barriers of rats. Embryotoxicity of baicalin should be considered as the key point to evaluate the safety of SHL.</p>

Study on Relationship between the sharing of HLA Gene Couples with Gestational Diabetes Mellitus / 中华围产医学杂志

Objective To assess the relationship between the HLA gene sharing of couples and gestational diabetes mellitus (GDM). Methods Thirty GDM women and their husbands served as study group, 40 normal pregnant women and their husbands were selected as control group. All pregnant women were primipara with single fetus. 5 ml cubital venous blood were taken. Genomic DNA was extracted using phenol chloroform method. HLA-DRB alleles type were determined with PCR-SSP. Results The sharing of HLA-DRB1 between couples in GDM group were 33.33%, significantly higher than that of normal pregnancy(12.50%)(P0.05). Conclusions The sharing of HLA-DRB1 between couples may inerease the risk of GDM, It suggested that placenta- and-fetus unit may play roles in GDM.