To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnH, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.

OBJECTIVETo study the effect of the HSP90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on cell proliferation and apoptosis of human cancer SGC-7901 cells and explore the mechanisms.

METHODSThe inhibitory effect of 17-AAG on the proliferation and morphology of SGC-7901 cells was assessed with MTT assay and DNA-PI staining, respectively. Flow cytometry was employed to analyze the changes in cell cycle and apoptosis of the cells following 17-AAG exposure. The cellular expression of Fas protein was detected by immunohistochemistry.

RESULTS17-AAG significantly suppressed the proliferation of SGC-7901 cells in a time- and dose-dependent manner. After treatment with 17-AAG for 48 h, SGC-7901 cells showed cell cycle arrested at G(2)/M stage, and the cell apoptosis rate increased with the 17-AAG concentration. The expression of Fas protein in the cytoplasm of SGC-7901 cells increased gradually with the increase of 17-AAG concentration.

CONCLUSION17-AAG can induce apoptosis, alters the cell cycle distribution and up-regulates the expression of Fas protein in SGC-7901 cells to suppress the cell proliferation.

Apoptosis ; drug effects ; Benzoquinones ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; pharmacology ; Stomach Neoplasms ; pathology ; fas Receptor ; metabolism

OBJECTIVE To study the effect of different concentrations of 17-AAG and EGCG monotherapy or in combination on the induced apoptosis in human nasopharyngeal carcinoma, and to explore new molecular targets for the treatment of nasopharyngeal carcinoma. METHODS MTT colorimetric method and fluorescent staining were used to detect the change of CNE proliferation inhibition rate and cell morphology. And furthermore, the expression level of Bcl-2, Bax, Caspase-3 were detected by RT-PCR. RESULTS 1. 17-AAG or EGCG alone had inhibitory effect on the human nasopharyngeal carcinoma CNE cells at 24 h, 48h and 72 h, and it was related with time and dose(P<0.01). The inhibition effect of combination of 17-AAG and EGCG was significantly increased,which was time and dose dependent(P<0.01). 2. RT-PCR was used to detect the mRNA expression level of Bcl-2, Bax and caspase-3. The level of Caspase-3 and Bax mRNA expression after treated by 17-AAG and EGCG was significantly higher, and the level of bcl-2 mRNA expression was lower than that after treated by 17-AAG or EGCG alone. CONCLUSION Our investigation implied that 17-AAG and EGCG in combination can effectively inhibit the proliferation of human nasopharyngeal carcinoma CNE cells. The involved mechanisms may be associated with the upregulation of Bax and Caspase-3 expression.

AIM:To explore the synergistic sensitization effect of human umbilical cord mesenchymal stem cell culture supernatant(hUMSC-CM)combined with temozolomide(TMZ)on various glioma cell lines,and to elucidate the underlying mechanisms.METHODS:The hUMSC-CM was harvested using two different serum deprivation tech-niques at 24 and 48 h,and was converted into freeze-dried powder,which was then given to rat malignant glioma cell line RG-2,human astrocytoma cell line U251 and human glioblastoma cell line LN-428 at 5 concentrations(0,1,3,6 and 9 g/L).The effectiveness and sensitivity of hUMSC-CM for inhibiting growth of glioma cells at 24,48 and 72 h were as-sessed using CCK-8 assay.Hematoxylin-eosin(HE)staining combined with CCK-8 assay was employed to evaluate the chemotherapy sensitivity of glioma cells after 48 h of treatment with TMZ at 6 concentrations(0,25,50,100,200 and 400 μmol/L).Two concentrations(3 and 9 g/L)of hUMSC-CM and 3 concentrations(50,100 and 200 μmol/L)of TMZ were chosen for concurrent treatment of glioma cells to assess the proliferation and pathological alterations.TUNEL staining was utilized to detect apoptosis.Flow cytometry was utilized to analyze cell cycle modifications.The expression alterations of apoptosis-inducing proteins,cleaved caspase-3,cleaved caspase-8 and cleaved PARP1,as well as autophagy-inducing proteins beclin-1 and LC3,were examined using Western blot to investigate the synergistic sensitization mechanism of hUMSC-CM combined with TMZ in vitro.RESULTS:The susceptibility of glioma cell lines to hUMSC-CM and TMZ varied,with RG-2 showing the highest sensitivity,followed by U251,and then LN-428.The inhibitory effect of hUMSC-CM(3 and 9 g/L)and TMZ(50,100 and 200 μmol/L)combined treatment on glioma cells was significantly greater than that that of single-agent treatments(P＜0.05),demonstrating a dose-and concentration-dependent enhancement.Notably,the combination of 9 g/L hUMSC-CM(C9)with 50 μmol/L TMZ(T50)effectively suppressed glioma cell growth.CCK-8 as-say indicated a significant reduction of cell viability in C9+T50 group compared with either C9 or T50 alone(P＜0.05).HE staining and TUNEL staining revealed pronounced morphological changes and significant apoptotic features in glioma cells treated with C9+T50.Flow cytometric analysis confirmed that C9+T50 induced cell cycle arrest in glioma cells.Fur-thermore,compared with control group,the levels of cleaved caspase-3,cleaved caspase-8,cleaved PARP1,beclin-1,and LC3-Ⅱ/LC3-Ⅰ were significantly elevated in the C9+T50-treated glioma cells(P＜0.01).CONCLUSION:(1)The concomitant administration of hUMSC-CM and TMZ exerts a broad inhibitory effect on glioma cells,with a synergistic sen-sitization observed across different cell lines.(2)The enhancement of glioma cell sensitivity to TMZ by hUMSC-CM may be attributed to the modulation of caspase-8/caspase-3/PARP1 signaling pathway and the induction of both apoptosis and autophagy in glioma cells.

Objective:To evaluate the prognostic value of thromboelastography maximum amplitude(MA)and arterial blood lactate levels for sepsis in elderly patients.Methods:A retrospective analysis was performed on clinical data of 63 sepsis patients(≥60 years old)admitted to the Intensive Care Unit(ICU)of Bethune Hospital of Shanxi Province from December 2018 to February 2020.MA values, white blood cell counts, lymphocyte counts, platelets, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)scores, sequential organ failure assessment(SOFA)scores, underlying diseases, body mass index, laboratory test results and other related treatments were analyzed.The subjects were divided into the survival group and the death group according to the 28-day survival outcome.Differences in MA, APACHE Ⅱ scores, SOFA scores and laboratory test results between the two groups were analyzed, and the correlations of MA with infection parameters and age were examined.Influencing factors of survival outcomes were analyzed using multivariate Logistic regression.The receiver operating characteristic curve(ROC)was used to calculate the prognostic value of MA and arterial lactate for sepsis in elderly patients.Results:The main sources of infections were pulmonary and abdominal(79.4%, 50/63)in 63 elderly patients with sepsis.The incidences of positive blood cultures and deaths were 15.9%(10/63)and 66.7%(42/63), respectively.There existed significant differences in lymphocyte counts, arterial lactate levels, MA and lengths of stay in the ICU between the survival group and the death group( t=3.847, 2.153, 2.745, -3.574, respectively, all P<0.05).MA was correlated with arterial lactate, SOFA score and survival outcome( r=-0.498, -0.506, and -0.358, respectively, all P<0.05).Multivariate Logistic regression analysis showed that MA and arterial lactate were independent factors for the survival outcome( OR=1.626, 0.766, all P<0.05).The area under the ROC curve(AUC, 95% CI)for the combination of MA and arterial lactate was larger than that of either MA or arterial lactate alone(0.89, range: 0.763-0.846; 0.58, range: 0.574-0.730; 0.77, range: 0.521-0.832; all P<0.05). Conclusions:The combination of thromboelastography maximum amplitude and lactate in arterial blood has important clinical value in assessing the prognosis of elderly patients with sepsis.