Aim To estimate the anti-uterine cervix cancer effects of the extracts from Lysimachia clethroides Duby in vivo and in vitro. Methods Inhibitory effect on growth was observed by MTT, 3H-TdR and colony-forming units assay. Apoptosis was detected by Hoechst fluorescent staining analysis. The effect on cell migration and morphology was observed by scratch test; Detecting effects of ZE4 on transplant tumor (U14) in mice through observing the tumor weight and organ index. Results ZE4 could inhibit the growth and migration of HeLa cell significantly and induce cell apoptosis. Its half inhibitory concentration (IC50) at 48h was 40.56 mg?L-1 by 3H-TdR assay. The inhibition rate of ZE4 against U14 was 49.9% at the dose of 400 mg?kg-1. Conclusions ZE4 could inhibit the growth of uterine cervix cancer in vitro and in vivo.

[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .

Objective To evaluate the effect of electroacupuncture at Chize and Shangjuxu on the prognosis and respiratory dynamics in patients with acute respiratory distress syndrome(ARDS).Methods A prospective randomized controlled study was conducted to collect patients with ARDS admitted to the Department of Emergency Medicine of Tongde Hospital of Zhejiang Province from December 2020 to September 2022.A total of 142 patients with ARDS from the Department of Emergency Medicine of Tongde Hospital of Zhejiang Province were selected for this study.Seven patients were excluded due to rapid deterioration,3 patients withdrew the informed consent,and finally 132 patients were enrolled.According to the order of enrollment,they were randomly divided into the electroacupuncture group and the conventional treatment group based on the random number table.The conventional treatment group received conventional treatment for ARDS,while the electroacupuncture group combined with electroacupuncture at Chize and Shangjuxu,the treatment lasted for 30 minutes.The mortality at day 60 and secondary efficacy indicators such as hospital stay,ventilator-free days at 28 day,and mechanical ventilation time of two groups were compared,and the indicators of respiratory mechanics and oxygenation such as inhalation oxygen concentration(FiO2),tidal volume(VT),positive end-expiratory pressure(PEEP),plateau pressure(Pplat),lung static compliance(Cst),oxygenation index(PaO2/FiO2)were also compared.Then the Kaplan-Meier survival curves were drawn to compare the differences in cumulative survival rates within 60 days between the two groups.Results The mortality at day 60 in the electroacupuncture group was lower than that in the conventional treatment group[33.33％(22/66)vs.40.91％(27/66)],but the difference was not statistically significant(P>0.05),and the hospital stay in the electroacupuncture group was significantly shorter than that in the conventional treatment group(days:25.09±11.57 vs.30.21±15.94,P<0.05).The ventilator-free days at 28 days in the electroacupuncture group were significantly higher than those in the conventional treatment group(days:10.06±7.35 vs.7.67±5.72,P<0.05).There was no statistically significant difference in mechanical ventilation time between the two groups(days:15.70±9.05 vs.18.39±9.55,P>0.05).After 72 hours of treatment,the FiO2,VT,and PaCO2 levels in the electroacupuncture group were significantly lower than those in the conventional treatment group[FiO2:0.39±0.07 vs.0.44±0.09,VT(mL/kg):6.27±0.74 vs.6.62±0.74,PaCO2(mmHg,1 mmHg≈0.133 kPa):39.94±4.52 vs.41.95±4.19,all P<0.05].Conclusion Electroacupuncture at Chize and Shangjuxu can effectively shorten the course of ARDS and improve prognosis,which may be related to the improvement of respiratory dynamics by electroacupuncture treatment.

Objective:To explore whether thyroxine (T 4) could promote differentiated thyroid cancer (DTC) progression by binding to integrin α vβ 3in vitro and its downstream mechanism. Methods:Papillary thyroid cancer cell lines TPC-1, K1 and follicular thyroid cancer (FTC) cell line FTC133 were cultured in vitro, and the expressions of integrin α vβ 3 in those 3 DTC cell lines were determined with immunofluorescence and flow cytometry analysis. After the treatment of T 4, tetraiodo thyroacetic acid (Tetrac) and Arg-Gly-Asp (RGD) peptide alone or in combination, the proliferation and metastatic potential of DTC cell lines were detected by cell counting kit-8 (CCK-8), Transwell migration and invasion assays. The small interfering RNA (siRNA) transfection was used to verify whether integrin α v or β 3 subunit knockdown could reverse the effect of T 4 on DTC cells. The expression levels of downstream signaling proteins phosphorylated extracellular signal-regulated kinase (p-ERK)1/2 and total extracellular signal-regulated kinase (ERK)1/2 were detected by Western blot. The effects of mitogen-activated protein kinase kinase (MEK)1/2 inhibitor (GSK1120212) on the proliferation, migration and invasion of T 4-treated cells were detected. One-way analysis of variance and Tukey test were used for data analysis. Results:The integrin α vβ 3 expressions in TPC-1, K1 and FTC133 cells were all positive, with the relative mean fluorescence intensity (MFI) of 61.93±18.61, 16.89±2.43 and 32.36±0.83, and the percentages of positive cells of (94.38±1.30)%, (74.11±3.87)% and (50.67±1.78)%, respectively ( F values: 13.36 and 217.30, P=0.006 and P<0.001). Compared with control group, the proliferation, migration and invasion in the three DTC cell lines treated with T 4 were significantly enhanced (96 h, F values: 62.67-297.50, q values: 13.15-20.73, all P<0.001). T 4-induced cell proliferation, migration and invasion were markedly reversed by Tetrac or RGD (96 h, q values: 8.61-17.54, all P<0.001). T 4-induced cell proliferation, migration and invasion were also significantly inhibited by the knockdown of integrin α v or β 3 subunit (72 h, F values: 7.75-70.98, q values: 4.77-15.21, all P<0.05). Western blot results showed that the phosphorylation levels of ERK1/2 in DTC cells were significantly increased by T 4 treatment, and the T 4-induced activation of ERK1/2 signaling pathway could be blocked by Tetrac, RGD, integrin α v or β 3 subunit knockdown. T 4-induced cell proliferation, migration and invasion were significantly reversed by GSK1120212 (96 h, F values: 47.53-151.40, q values: 10.32-16.65, all P<0.001). Conclusion:T 4 can promote cell proliferation and metastasis of DTC cells by binding to integrin α vβ 3 and activating the ERK1/2 pathway.

Objective: To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function. Methods: Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS-PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr-activated SepharoseTM 4B. Reverse high pressure liquid chromatography (HPLC), SDS-PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone-antigen peptide complexes. Results: The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70-antigen peptide and 1.00 μg HSP90-antigen peptide activated lymphocytes significantly. Their A₄₉₀ values were 0.26±0.03, 0.45±0.05 and 0.32±0.04, respectively, while the corresponding doses of HSP60-antigen peptide and HSP110-antigen peptide did not activate lymphocytes. The killing rates of 1.00 μg HSP70-antigen peptide and 1.00 μg HSP70 were (45.0±2.0)% and (16.0±2.0)%, respectively, showing a significant difference (P=0.012). Similarly, the killing rates of 1.00 μg HSP90-antigen peptide and 1.00 μg HSP90 were (36.0±5.0)% and (13.0±4.0)%, respectively, also showing a significant difference (P=0.048). Conclusions The amount of chaperone antigen peptide complexes in gastric cancer cells is extremely low, but it is obviously increased in gastric cancer stem cells. After purification, the chaperone antigen peptide complexes with high purity can be prepared. The extracted chaperone antigen peptide complexes have stronger immunogenicity, and can be used to make tumor vaccine in vitro, which may have a good application value in the targeted therapy of gastric cancer.