Effect of Baicalin on Pyroptosis of Diffuse Large B-Cell Lymphoma Cell Lines DB and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of Baicalin on the proliferation and pyroptosis of diffuse large B-cell lymphoma cell line DB and its mechanism.@*METHODS@#DB cells were treated with baicalin at different concentrations (0, 5, 10, 20, 40 μmol/L). Cell proliferation was detected by CCK-8 assay and half maximal inhibitory concentration (IC50) was calculated. The morphology of pyroptosis was observed under an inverted microscope, the integrity of the cell membrane was verified by LDH content release assay, and the expressions of pyroptosis-related mRNA and protein (NLRP3, GSDMD, GSDME, N-GSDMD, N-GSDME) were detected by real-time fluorescence quantitative PCR and Western blot. In order to further clarify the relationship between baicalin-induced pyroptosis and ROS production in DB cells, DB cells were divided into control group, baicalin group, NAC group and NAC combined with baicalin group. DB cells in the NAC group were pretreated with ROS inhibitor N-acetylcysteine (NAC) 2 mmol/L for 2 h. Baicalin was added to the combined treatment group after pretreatment, and the content of reactive oxygen species (ROS) in the cells was detected by DCFH-DA method after 48 hours of culture.@*RESULTS@#Baicalin inhibited the proliferation of DB cells in a dose-dependent manner (r=-0.99), and the IC50 was 20.56 μmol/L at 48 h. The morphological changes of pyroptosis in DB cells were observed under inverted microscope. Compared with the control group, the release of LDH in the baicalin group was significantly increased (P<0.01), indicating the loss of cell membrane integrity. Baicalin dose-dependently increased the expression levels of NLRP3, N-GSDMD, and N-GSDME mRNA and protein in the pyroptosis pathway (P<0.05). Compared with the control group, the level of ROS in the baicalin group was significantly increased (P<0.05), and the content of ROS in the NAC group was significantly decreased (P<0.05). Compared with the NAC group, the content of ROS in the NAC + baicalin group was increased. Baicalin significantly attenuated the inhibitory effect of NAC on ROS production (P<0.05). Similarly, Western blot results showed that compared with the control group, the expression levels of pyroptosis-related proteins was increased in the baicalin group (P<0.05). NAC inhibited the expression of NLRP3 and reduced the cleavage of N-GSDMD and N-GSDME (P<0.05). Compared with the NAC group, the NAC + baicalin group had significantly increased expression of pyroptosis-related proteins. These results indicate that baicalin can effectively induce pyroptosis in DB cells and reverse the inhibitory effect of NAC on ROS production.@*CONCLUSION@#Baicalin can inhibit the proliferation of DLBCL cell line DB, and its mechanism may be through regulating ROS production to affect the pyroptosis pathway.

Regulation of Baicalin on Growth of Extranodal NK/T Cell Lymphoma Cells through FOXO3/CCL22 Signaling Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of baicalin on the growth of extranodal NK/T cell lymphoma (ENKTCL) cells and its related mechanism.@*METHODS@#Normal NK cells and human ENKTCL cells lines SNK-6 and YTS were cultured, then SNK-6 and YTS cells were treated with 5, 10, 20 μmol/L baicalin and set control. Cell proliferation and apoptosis was detected by Edu method and FCM method, respectively, and expressions of BCL-2, Bax, FOXO3 and CCL22 proteins were detected by Western blot. Interference plasmids were designed and synthesized. FOXO3 siRNA interference plasmids and CCL22 pcDNA overexpression plasmids were transfected with PEI transfection reagent. Furthermore, animal models were established for validation.@*RESULTS@#In control group and 5, 10, 20 μmol/L baicalin group, the proliferation rate of SNK-6 cells was (56.17±2.96)%, (51.92±4.63)%, (36.42±1.58)%, and (14.60±2.81)%, respectively, while that of YTS cells was (58.85±2.98)%, (51.38±1.32)%, (34.75±1.09)%, and (15.45±1.10)%, respectively. In control group and 5, 10, 20 μmol/L baicalin group, the apoptosis rate of SNK-6 cells was (5.93±0.74)%, (11.78±0.34)%, (28.46±0.44)%, and (32.40±0.37)%, respectively, while that of YTS cells was (7.93±0.69)%, (16.29±1.35)%, (33.91±1.56)%, and (36.27±1.06)%, respectively. Compared with control group, the expression of BCL-2 protein both in SNK-6 and YTS cells decreased significantly (P<0.001), and the expression of Bax protein increased in SNK-6 cells only when the concentration of baicalin was 20 μmol/L (P<0.001), while that in YTS cells increased in all three concentrations(5, 10, 20 μmol/L) of baicalin (P<0.001). The expression of FOXO3 protein decreased while CCL22 protein increased in ENKTCL cell lines compared with human NK cells (P<0.001), but the expression of FOXO3 protein increased (P<0.01) and CCL22 protein decreased after baicalin treatment (P<0.001). Animal experiments showed that baicalin treatment could inhibit tumor growth. The expression of CCL22 protein in ENKTCL tissue of nude mice treated with baicalin decreased compared with control group (P<0.01), while the FOXO3 protein increased (P<0.05). In addition, FOXO3 silencing resulted in the decrease of FOXO3 protein expression and increase of CCL22 protein expression (P<0.01, P<0.001).@*CONCLUSION@#Baicalin can inhibit proliferation and promote apoptosis of ENKTCL cell lines SNK-6 and YTS, up-regulate the expression of Bax protein, down-regulate the expression of BCL-2 protein, and down-regulate the expression of CCL22 protein mediated by FOXO3. Animal experiment shown that the baicalin can inhibit tumor growth. Baicalin can inhibit the growth and induce apoptosis of ENKTCL cells through FOXO3/CCL22 signaling pathway.

Changes of common pathogens of viral diarrhea in children during the COVID-19 epidemic / 中华临床感染病杂志

Objective:To analyze the changes of common pathogens of viral diarrhea in children during the COVID-19 epidemic.Methods:A retrospective analysis was performed on children with acute diarrhea treated in Children’s Hospital affiliated to Zhejiang University School of Medicine from 2019 April to 2019 December(before COVID-19, n=407)and 2020 April to 2020 December (during COVID-19, n=645). Children were further divided into 1-6 months, >6 months-2 years, >2-5 years age groups. Detection rates of norovirus GI (NOVI), norovirus GⅡ (NOVⅡ), rotavirus A (ROVA), rotavirus C (ROVC), intestinal adenovirus (EADV), sapovirus (SAV) and astrovirus (ASV) in different age groups and seasons before and during COVID-19 were compared by χ2 or Fisher exact test. Results:The total detection rates of diarrhea-causing viruses among age groups were significantly higher before COVID-19 than those during COVID-19( χ2=8.43, 38.22 and 9.23, all P<0.05). The detection rates of NOVⅡ and EADV in infants aged 1-6 months and >6 months-2 years were decreased during the period of COVID-19 epidemic( χNOVⅡ2 =36.87 and 17.77, both P<0.001, χEADV2 =9.08, P=0.014 and 0.003); the detection rates of NOVⅡ in children aged 2-5 years was decreased during the period of COVID-19 epidemic( χ2=3.96, P=0.047); the detection rates of other diarrhea-causing viruses were not decreased during the period of COVID-19 epidemic(all P>0.05). The detection rates of diarrhea-causing viruses among 4-6 months, 7-9 months and 10-12 months were higher during the period of COVID-19 epidemic( χ2=11.62, 65.41 and 27.80, all P<0.001). Conclusion:After the outbreak of COVID-19, the detection rate of common pathogens of viral diarrhea in children is decreased, especially NOVⅡ and EADV, which may be related to the measures in response to major public health events in Zhejiang Province.

Preparation and characterization of glycyrrhetinic acid-modified nano graphene oxide drug delivery system / 中国中药杂志

In this study,a nano drug delivery system GA-DTX-NGO which could be used for liver tumor photothermal and chemotherapy was prepared and characterized,with docetaxel(DTX) as model drug,glycyrrhetinic acid(GA) as the target molecule,and nano graphene oxide(NGO) as the photosensitizer. Firstly,GA-NGO nanocomposites were synthesized by the amidation reaction,and then GA-DTX-NGO was prepared by ultrasonic dispersion method. The encapsulation efficiency and drug loading ratio were determined by high performance liquid chromatography(HPLC) and ultracentrifugation; the morphology was observed by transmission electron microscopy(TEM). The photothermal conversion test was carried out by laser irradiation at 808 nm and the drug release test in vitro was performed using reverse dialysis. Finally,the effect of GA-DTX-NGO on SMMC-7721 liver tumor cells proliferation was determined by using MTT assay. The results showed that GA-DTX-NGO had good water dispersibility,and TEM results showed a lamellar structure with about 200 nm in diameter. The encapsulation efficiency and drug loading ratio of GA-DTX-NGO were(98. 89 ± 0. 07) % and(64. 74±0. 26) %,respectively. GA-DTX-NGO had strong photothermal conversion performance under 808 nm of laser irradiation. The drug release test in vitro results showed GA-DTX-NGO had obvious sustained-release effects and temperature-dependent release characteristics. The results of cell assay showed that GA-DTX-NGO could effectively inhibit the proliferation of SMMC 7721 cells in a concentration-and time-dependent manner,and the inhibitory effect was enhanced after combination with the near-infrared therapy. In conclusion,the preparation process of GA-DTX-NGO nano drug delivery system is feasible,which could provide some theoretical basis for further study of photothermal and chemotherapy on liver tumor.

The expression levels of TNF-α and IL-6 in tumor tissues and change of serum ferritin concentration in patients with intracranial aneurysm and its clinical significance / 中国基层医药

Objective To investigate the expression levels of tumor necrosis factor alpha (TNF-α),interleukin-6(IL-6) and the change of serum ferritin in patients with intracranial aneurysm and its clinical significance.Methods 22 patients with intracranial aneurysm and 16 cases of traumatic brain injury who recevied operation in Department of Neurosurgery in Quzhou People's Hospital from Jan.2014 to Jan.2015 were enrolled as observation group and control group.The intracranial aneurysm tissues and normal cerebral vascular tissues were collected respectively to detect the situation of inflammatory cell infiltration,and the expression of TNF-α and IL-6 was detected by HE staining and immunohistochemical staining.The serum was collected on hospitalized,postoperative 3 days and postoperative 7 days to detect serum ferritin.Results The positive cells were observed in the observation group after immunohistochemical staining,in which the expression levels of TNF-α [(0.194 ± 0.074) vs.(0.135 ± 0.047),t =2.799] and IL-6 [(0.152 ± 0.057) vs.(0.103 ± 0.028),t =3.494] were significantly higher than those in the control group(P =0.008,0.001).The serum ferritin level in the observation group was significantly higher than that in the control group at postoperative 3d [(232.25 ± 105.26) ng/mL vs.(169.51 ± 66.24) ng/mL,t =2.097] and postoperative 7d[(263.39 ± 114.73) ng/mL vs.(166.57 ± 73.71) ng/mL,t =2.955] (P =0.043,0.005).The difference of serum ferritin on different day in the observation group also was statistically significant(F =8.625,P =0.003).Conclusion The expression levels of TNF-α and IL-6 in intracranial aneurysms were high,which may be one of the important factors in the formation of intracranial aneurysms,and it provides an important reference of early prevention and drug treatment for patients with intracranial aneurysm.The serum ferritin is related to the rupture and hemorrhage of intracranial aneurysm,which can be used to judge prognosis and guide rational treatment,and worth further study to confirm.