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ObjectiveTo investigate effect of lyophilized powder of modified Huangqi Gancaotang on proliferation， apoptosis， invasion， migration， and epithelial-mesenchymal transition of human non-small cell lung cancer cells （A549， PC9） and possible mechanism. MethodEffect of 1.0， 2.0， 4.0， 8.0， 12.0 g·L-1 modified Huangqi Gancaotang on the proliferation of non-small cell lung cancer cells was detected by cell counting kit-8 （CCK-8） assay. A549 and PC9 cells were classified into the blank group and the low-， medium-， and high-dose Huangqi Gancaotang groups （2.0， 4.0， 8.0 g·L-1）. Plate cloning assay was used to examine the effect of modified Huangqi Gancaotang on cell cloning ability. Hoechst 33342 staining and flow cytometry were employed to detect the apoptosis， and scratch assay and Transwell migration assay were applied to examine cell migration and invasion abilities， respectively. Mammosphere assay was used to examine the sphere-forming ability of tumor cells， and real-time polymerase chain reaction （Real-time PCR） to detect the mRNA expression of stemness-related molecules octamer-binding transcription factor 4 （Oct-4）， human sex-determining region Y-box 2 （Sox2）， and homeobox transcription factor （Nanog） to assess cancer stem cell activity. The protein expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated death promoter （Bad）， Bcl-2-associated X protein （Bax）， cleaved Caspase-3， Caspase-3， E-cadherin， N-cadherin， vimentin， matrix metalloproteinase-2 （MMP-2）， β-catenin， c-Myc， Cyclin D1， and zinc-finger transcription factor （Slug） was determined by Western blot. ResultThe proliferation ability of A549 and PC9 cells was significantly inhibited after 24 h and 48 h treatment with 1.0， 2.0， 4.0， 8.0， and 12.0 g·L-1 lyophilized powder of modified Huangqi Gancaotang compared with that in the blank group and the inhibition was dose- and time-dependent （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang suppressed the cloning ability of A549 and PC9 cells （P< 0.05， P<0.01）， and high-dose modified Huangqi Gancaotang induced apoptosis of A549 and PC9 cells （P<0.01）. In comparison with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang inhibited the invasion and migration of A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the low-， medium-， and high-dose modified Huangqi Gancaotang significantly decreased volume of the microspheres of A549 cells and the mRNA expression of Oct-4， Sox2， and Nanog in A549 and PC9 cells （P<0.05， P<0.01）. Compared with the blank group， the medium- and high-dose modified Huangqi Gancaotang down-regulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01）， up-regulated the expression of pro-apoptotic proteins Bad， Bax， and cleaved Caspase-3/Caspase-3 （P<0.05， P<0.01） in A549 and PC9 cells， decreased the expression of MMP-2， N-cadherin， and vimentin （P<0.05， P< 0.01）， and raised the E-cadherin expression （P<0.05， P<0.01）. Moreover， the medium-dose and high-dose modified Huangqi Gancaotang all reduced the expression of β-catenin， c-Myc， Cyclin D1， and Slug in A549 and PC9 cells （P<0.01）. ConclusionModified Huangqi Gancaotang can inhibit the proliferation， invasion， migration， activity of cancer stem cells， and epithelial-mesenchymal transition of human non-small cell lung cancer （A549， PC9） cells and induce apoptosis， and the mechanism is the likelihood that it regulates Wnt/β-catenin signaling pathway.
ABSTRACT
@#Objective To explore the surgical technique and preliminary results of endoscopic nipple-sparing mastectomy (E-NSM) and immediate pre-pectoral implant-based breast reconstruction (BR) with titanium-coated polypropylene mesh (TiLoop Bra) via single axillary incision for breast cancer patients. Methods The clinical data of 9 consecutive female patients who underwent E-NSM and immediate pre-pectoral implant-based BR with TiLoop Bra from March to May 2021 were retrospectively analyzed. The mean age of patients was 40.6 (22-60) years. The operation time, early complications were collected, and the patients' social and mental health, breast satisfaction and chest function before and after the operation were assessed with the BREAST-Q questionnaire. Results All the patients had unicentric tumor with a mean diameter of 2.4 (0.6-4.7) cm. The mean distance from the tumor to the nipple was 2.5 (2-4) cm. There were 2 patients with tumor stage 0 and 7 patients with stageⅠ. The mean operation time was 161.1 (125-201) min, the mean blood loss was 41.1 mL and the hospital stay time was 1.5 d. There were 5 patients in the day-care unit. All the patients were successfully followed up with a median follow-up time of 1 (1-2) month. One (11.1%) patient with depigmentation of the nipple-areola complex caused by mild ischemia. None of the patients had incision complications, subcutaneous emphysema, hematoma, infection, nipple-areola or skin flaps necrosis, implant loss. During the follow-up period, no local/regional recurrence or distant metastasis was found. Chest well-being was decreased in the first month after the surgery compared with preoperative status, and the difference was statistically significant (P=0.001). There was no statistical difference in the breast satisfaction or psychosocial function scores between pre- and post-operation (P>0.05). Conclusion E-NSM and immediate pre-pectoral implant-based BR with TiLoop Bra via single axillary incision has minimal trauma, rapid postoperative recovery, short operation time, few early complications and good early cosmetic effect, and the short-term result is satisfactory.
ABSTRACT
A gas chromatography mass spectrometry (GC-MS) method has been developed and fully validated for the simultaneous determination of natural borneol (NB) and its metabolite, camphor, in rat plasma. Following a single liquid-liquid extraction, the analytes were separated using an HP-5MS capillary column (0.25 mm ? 30 m ? 0.25μm) and analyzed by MS in the selected ion monitoring mode. Selected ion monitor (m/z) of borneol, camphor and internal standard was 95, 95 and 128, respectively. Linearity, accuracy, precision and extraction recovery of the analytes were all satisfactory. The method was successfully applied to pharmacokinetic studies of NB after oral administration to Wistar rats.