Classification and Identification of Cynomorii Herba from Different Producing Areas Based on Fourier Rransform Infrared Spectroscopy and Chemometrics / 中国实验方剂学杂志

Objective: To realize the classification and identification of Cynomorii Herba from different producing areas based on fourier transform infrared spectroscopy (FTIR) and chemometrics. Method: FTIR spectrum data of 106 batches of Cynomorii Herba from 12 cities in 5 provinces were collected by transmission method and preprocessed. The FTIR fingerprints of Cynomorii Herba were established, and spectrum analysis was performed. The FTIR similarities of Cynomorii Herba from different producing areas were calculated by correlation coefficient method. The first derivative (1D) spectrum of average FTIR of Cynomorii Herba from different producing areas were obtained. The soft independent modeling of class analog (SIMCA) model based on principal component analysis (PCA) was established by the preprocessed 1D spectrum data. The orthogonal partial least squares (OPLS) model was established by top 6 principal components. Result: The FTIR fingerprint trend and main absorption peaks of Cynomorii Herba from different producing areas were basically the same,and 16 common characteristic absorption peaks were recognized. Similarity and 1D spectrum of FTIR fingerprint of Cynomorii Herba from different producing areas showed significant and unique characteristics. The established SIMCA model can realize the classification and identification of Cynomorii Herba from different provinces,while OPLS model can realize accurate classification and identification of Cynomorii Herba in different cities. The classification and identification of Cynomorii Herba from 12 city producing areas showed obvious geographical clustering characteristics. Conclusion: The established method based on FTIR and chemometrics can realize the classification and identification of Cynomorii Herba from 12 cities.

Identification of Hedysari Radix from 8 Producing Counties in Gansu Province Based on FT-IR Fingerprint Characteristics / 中国中医药信息杂志

Objective To study the FT-IR fingerprint characteristics of Hedysari Radix from 8 producing counties in Gansu Province; To provide references for identification and application of Hedysari Radix in different producing counties. Methods FT-IR fingerprints of 110 batches of Hedysari Radix from 8 producing counties in Gansu Province were collected in the wave number range of 4000–400 cm-1. The common pattern of the fingerprints were analyzed, and the similarity analysis were used to analyze the FT-IR fingerprints of Hedysari Radix from 8 producing counties. The FT-IR fingerprint characteristics of Hedysari Radix from 8 producing counties in Gansu Province were compared. Results The rank of average similarity of FT-IR fingerprints of Hedysari Radix from 8 producing counties was Tanchang County > Li County > Xihe County > Wudu District > Zhang County > Min County > Longxi County >Weiyuan County, and Hedysari Radix from Longxi County and Weiyuan County were very different from other producing counties. The FT-IR fingerprints of Hedysari Radix from Longnan City (Tanchang County, Li County, Xihe County and Wudu District) were similar, and the average similarity was relatively high; while that from Dingxi City (Zhang County, Min County, Longxi County and Weiyuan County) were similar, and the average similarity was relatively low. Hedysari Radix from every producing county had a significant and unique FT-IR fingerprint characteristic. Conclusion The identification and application of Hedysari Radix from 8 producing counties in Gansu Province can be realized according to FT-IR fingerprint characteristics.

Proto-oncogene c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways.

BACKGROUND: c-erbB2, a proto-oncogene coding epidermal growth factor receptor-like receptor, also as a chemosensitivity/prognosis marker for gynecologic cancer, may be involved in initiation of growth of rat primordial follicles. The aim of the present study is to investigate the role and signal pathway of c-erbB2 in onset of rat primordial follicle development. METHODS: The expression of c-erbB2 mRNA and protein in neonatal ovaries cultured 4 and 8 days with/without epidermal growth factor (EGF) were examined by in situ hybridization, RT-PCR and western blot. The function of c-erbB2 in the primordial folliculogenesis was abolished by small interfering RNA transfection. Furthermore, MAPK inhibitor PD98059 and PKC inhibitor calphostin were used to explore the possible signaling pathway of c-erbB2 in primordial folliculogenesis. RESULTS: The results showed that c-erbB2 mRNA was expressed in ooplasm and the expression of c-erbB2 decreased after transfection with c-erbB2 siRNA. Treatment with EGF at 50 ng/ml significantly increased c-erbB2 expression and primary and secondary follicle formation in ovaries. However, this augmenting effect was remarkably inhibited by c-erbB2 siRNA transfection. Furthermore, folliculogenesis offset was blocked by calphostin (5 x 10(-4) mmol/L) and PD98059 (5 x 10(-2) mmol/L), but both did not down-regulate c-erbB2 expression. In contrast, the expressions of p-ERK and p-PKC were decreased obviously by c-erbB2 siRNA transfection. CONCLUSIONS: c-erbB2 initiates rat primordial follicle growth via PKC and MAPK pathways, suggesting an important role of c-erbB2 in rat primordial follicle initiation and development.

Roles of proto-oncogene c-erbB2 during the initiation growth of rat primordial follicles / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the expression and possible roles of proto-oncogene c-erbB2 during the initiation growth of primordial follicles.</p><p><b>METHODS</b>Ovaries were collected from 2-day-old SD rats and cultured in the Waymouth culture system. In-situ hybridization, RT-PCR and immunohistochemistry were performed to assess the expressions of c-erbB2 mRNA and protein during the initiation growth of primordial follicles and after the effect of EGF. Western blot was used to observe the PCNA, p-ERK1/2 contents and correlation analysis was used to study the correlation relationship between contents of p-ERK1/2 and expressions of c-erbB2 mRNA at the same time of the primordial follicles growth.</p><p><b>RESULTS</b>PCNA protein levels appeared to be more intense during the initiation growth of primordial follicles, EGF could promote the proliferation and differentiation of the primordial follicles. c-erbB2 mRNA existed in the oocytes endochylema and ErbB2 existed in the oocytes membrane, the expressions of c-erbB2 mRNA and ErbB2 appeared to be more intense when primordial follicles were cultured for 8 d than cultured for 0 d in the Waymouth culture system and were further increased with 50 ng/ml EGF for 4 d and 8 d. The same results were observed by RT-PCR, too. p-ERK1/2 protein levels were consistent with the changes of c-erbB2 mRNA and protein. Furthermore, Spearman rank correlation analysis showed there was a significant positive correlation relationship between the changes of p-ERK1/2 and the changes of c-erbB2 mRNA during the primordial follicles growth and after the effect of EGF (rs = 0.900, P < 0.05).</p><p><b>CONCLUSION</b>It was suggested that proto-oncogene c-erbB2 may be play an important role during the initiation growth of primordial follicles with EGF, and it is indirectly suggested that c-erbB2 promotes the development of the primordial follicles via ERK-MAPK signal transduction.</p>

The expression of proto-oncogene c-erbB₂ and its role in the initiation of primordial follicle growth in rat ovary / 生理学报

Little is known about the factors that control the initiation of growth of primordial follicles. The objective of the present study was to investigate the effect of c-erbB₂ on the onset of primordial follicle development, and whether c-erbB₂ mediates the effect of epidermal growth factor (EGF) in this process. We synthesized three pairs of siRNAs targeting the c-erbB₂ mRNA and transferred them into the newborn rat ovary cultured in vitro with Metafectene. After siRNAs transfection, the efficiency of siRNAs was tested by examining c-erbB₂ mRNA and protein levels. The level of c-erbB₂ mRNA was reduced by 49.6%, 46.7% and 82.6% respectively after transfecting siRNA1, siRNA2 and siRNA3, and the level of ErbB₂ protein also reduced remarkably after siRNA3 transfection. c-erbB₂ siRNA3 significantly inhibited the primordial follicle initiation and development; EGF augmented primordial follicles formation, but the effect was abolished by c-erbB₂ siRNA3. All of these results suggest that c-erbB₂ plays an important role in primordial follicle development and folliculogenesis initiation, and mediates the effect of EGF on primordial follicle development.

Role of MAPK and PKC signaling pathways in the regulation of c-erbB₂ in the primordial follicles onset / 生理学报

Our previous studies showed that the proto-oncogene c-erbB₂ played an important role in primordial follicles growth. The present study was conducted to investigate the role of MAPK and PKC signaling pathways in the primordial follicle onset in neonatal rats, and the relationship between c-erbB₂ and MAPK/PKC signaling pathways. Ovaries collected from 2-day-old Sprague-Dawley rats were cultured in the Waymouth culture system in vitro. Ovaries were transfected with c-erbB₂ siRNA, or treated with PD98059 (50 mumol/L) or Calphostin (0.5 mumol/L) in the culture medium. RT-PCR was performed to measure the expression of c-erbB₂ mRNA, and Western blot analysis was performed to measure the expression of ErbB₂, MAPK and PKC protein after the neonatal rat ovaries were cultured for 8 d. The quantities of every-stage follicles of ovaries cultured for 8 d were obtained in histological section stained with hematoxylin eosin. The results showed that c-erbB₂ siRNA reduced the levels of c-erbB₂ mRNA (P<0.01) and the levels of ErbB₂, MAPK and PKC protein (P<0.01) significantly. But the levels of c-erbB₂ mRNA and ErbB₂ protein exhibited no change (P>0.05) in the ovaries cultured with PD98059 or Calphostin. After the ovaries were transfected with c-erbB₂ siRNA or cultured with PD98059 or Calphostin for 8 d, the quantities of primary follicles and second follicles were lower than those in the control group (P<0.05 or P<0.01), but the quantity of the primordial follicles was higher than that in the control group (P<0.01). These results suggest that proto-oncogene c-erbB₂ promotes the initiation of primordial follicle growth through the MAPK and PKC signal transduction, and c-erbB₂ is possibly the upstream of PKC and MAPK signaling pathway in the regulation of primordial follicle onset.