OBJECTIVE:To investigate inhibitory effects of matrine on the proliferation of human bladder cancer BIU-87 cells and its mechanism. METHODS:The cell viability was detected by MTT assay and inhibitory rate of cell proliferation was calculat-ed after human bladder cancer BIU-87 cells were treated with 0(negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 24,48 and 72 h,respectively. After treated with 0 (negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 48 h,the cell cycle and apoptotic rate were detected by flow cytometry;the expression of Survivin,Caspase-3 and Caspase-7 protein were detected by Western blot assay. RESULTS:Compared with negative control,the proliferation of BIU-87 cells were significantly inhibited after incubated with 1.0-4.0 mg/ml matrine for 24,48 and 72 h(P<0.05 or P<0.01),and inhibitory rate of cell proliferation increased in concentration and time-dependant manner;after treated for 48 h,the percentage of G0/G1 phase cells and apoptotic rate in-creased,while the percentage of cells at S phase and G2/M phase were decreased (P<0.05 or P<0.01);the expression of Cas-pase-3 and Caspase-7 protein increased,while the expression of survivin protein decreased after incubated with 0.5-4.0 mg/ml ma-trine for 48 h. CONCLUSIONS:Matrine can inhibit the proliferation of human bladder cancer BIU-87 cells,block the cell cycle and induce apoptosis;its mechanism may be related to the expression regulation of Survivin,Caspase-3 and Caspase-7.

Objective To investigate the effects of ligustilide on the proliferation of human lung adenocarcinoma A549 cells and its mechanism. Methods CCK-8 method was used to detect the effects of ligustilide on activity of A549 cells. Apoptosis rate was measured by TUNEL. Nuclear morphological changes of A549 cells were observed by fluorescence microscope after staining by Hoechst 33258. ELISA was used to detect the VEGF levels after incubation with ligustilide. Western blot was used to detect the expression of NF-κB, Bcl-2, Bax and the protein expression of phosphorylation of JNK. Results A549 cell activity was significantly inhibited after incubated with 15, 30, 60, 120, 180μmol/L ligustilide for 12, 24, 48 h (P<0.05, P<0.01, P<0.001) in a dose and time-dependent manner (P<0.05, P<0.01);Apoptosis rate increased by 30, 60, 120 μmol/L ligustilide for 12, 24, 48 h in a dose and time-dependent manner;A549 cells treated with ligustilide for 48 h appeared the apoptosis forms, including nuclear shrinkage, beating, strong blue fluorescence staining, and the chromatin divided producing apoptotic body;30, 60, 120μmol/L ligustilide increased the expression of p65 subunit of NF-κB protein in the nuclei and the phosphorylation levels of JNK protein, reduce the expression of suppression apoptosis protein Bcl-2, and raised the expression of pro-apoptotic protein Bax (P<0.05, P<0.01, P<0.001). Conclusion By inhibiting the expression of VEGF, ligustilide can inhibit the formation of tumor angiogenesis. By regulating the expression of NF-κB and the phosphorylation levels of JNK protein and reducing the ratio of Bcl-2/Bax, ligustilide can promote tumor cell apoptosis.

Objective To investigate the proliferation inhibitory effect and mechanism of Ginkgo Biloba extract (EGB) on non-small cell lung cancer (NSCLC) PC-9 cells. Methods Concentrations of EGB were set as 70 mg/L, 105 mg/L, 140 mg/L, 210 mg/L, and 280 mg/L, and were used to culture PC-9 cells in vitro for 24 h, 48 h, and 72 h. Tetramethylazo thiazole blue staining (MTT) method was applied to detect cell inhibition rate. Hoechst33258 fluorescent staining was employed to observe the nucleus. SOD activity and MDA content were detected by ELISA. The protein expressions of Caspase-3, Caspase-9, Cyt-C, and AIF were detected by Western blot. Results After incubated for 24 h, 48 h, and 72 h, EGB inhibited the proliferation of PC-9 cells, IC50 was 195.45 mg/L, 179.63 mg/L, 142.23 mg/L, respectively). 105 mg/L, 140 mg/L, 210 mg/L EGB could induce apoptosis of PC-9 cells, cause the nucleus pycnosis, produce apoptotic bodies, improve SOD activity and decrease MDA content (P<0.05, P<0.01, P<0.001). Western blot results showed that, compared with the control group, EGB can obviously increased the protein expressions of Caspase-3, Caspase-9, Cyt-C, and AIF, with statistical significance (P<0.05, P<0.01, P<0.001). Conclusion EGB can effectively restrain proliferation of non-small cell lung cancer PC-9 cells, the mechanism may be realized by inducing PC-9 cell apoptosis through pathway of mitochondria apoptosis, reducing oxidation, and clearing free radicals.

Objective To investigate the effects of extract of Ginkgo biloba leaves EGb761 on 1-methy-l 4-phenylpyridium (MPP+)-induced injury in human neuroblastoma SH-SY5Y cells; To discuss its mechanism of action.Methods Cell culture method was used and SH-SY5Y cell damage model was induced with different concentrations of MPP+ to build Parkinson’s disease model in vitro. The experiment was divided into control group, MPP+ model group, low-, medium-, and high-dose EGb761 groups. The survival rate was determined by MTT assay, and the apoptotic rate was detected by flow cytometry according to AnnexinV apoptosis detection kit. The cell morphology was observed by inverted microscope. NO content in SH-SY5Y cells was detected by Nitric acid reduction method.Results Compared with the control group, the survival rate of SH-SY5Y cells decreased and the apoptotic rate and NO content increased in the model group (P<0.05); Compared with the model group, the survival rate of SH-SY5Y cells increased and the apoptotic rate and NO content decreased in the low-, medium- and high-dose EGb761 groups (P<0.05).Conclusion EGb761 can protect MPP+-induced SH-SY5Y cell from damage by the inhibition of the content of NO free radical.

OBJECTIVETo investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model.

METHODSMice were randomly assigned to a control group and a hepatic fibrosis model group. The control group was further divided into three subgroups for use as normal controls (A1), mineral oil-treated controls (A2), and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1), 10-weeks models (B2), low-dose (L)-FZHc models (C1), and high-dose (H)-FZHc models (C2). The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3, C1, and C2 starting at week 7 of the experiment. At the end of week 6 and 10, hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining. Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2, NAD(P)H quinine oxidoreductase 1 (Nqol), a-smooth muscle actin (a-SMA) and fibronectin (FN). Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression. Western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation. F test, LSD test and ridit test were used for statistical analyses.

RESULTSCompared with the B2 group (ridit value: 0.09), the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group, ridit value: 0.32) and high doses (C2 group, ridit value: 0.40) (F =82.927, P less than 0.05). In addition, compared with the B2 group, the model groups treated with FZHc showed significantly decreased expression of a-SMA and FN proteins, with a dose-dependent trend (by immunohistochemistry: C 1 group at the end of 10 weeks, F =77.421, 118.262, P less than 0.05; C2 group, P =0.002, 0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistry:C1 and C2 groups at the end of 10 weeks, F =182.537, 75.615, P less than 0.05 and by westen blotting: F =45.664, 127.673, P less than 0.05), which also showed a dose-dependent trend (C2 group, P =0.000, 0.014; 0.005, 0.014). Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs. B2 group, F =94.787, P less than 0.05), and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs. C1 group, P =0.044). Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105, P less than 0.05), and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094).

CONCLUSIONFuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport, as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN. Therefore, Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hepatocytes ; drug effects ; metabolism ; Liver Cirrhosis, Experimental ; metabolism ; Mice ; Mice, Inbred Strains ; NAD(P)H Dehydrogenase (Quinone) ; metabolism ; NF-E2-Related Factor 2 ; metabolism

【Objective】 To analyze the impact of sporadic cases of COVID-19 on the work of Transfusion Department, so as to explore the countermeasures. 【Methods】 The admission of inpatient departments, the reception of outpatient(including emergency) departments, the workload of transfusion department(including blood typing, unexpected antibody screening and cross matching), and the consumption of blood components in the Xijing Hospital between October and November in 2021, during COVID-19 outbreak, were collected. All the above data was statistically compared to the data in same period in 2018, before the COVID-19 outbreak. 【Results】 Due to the COVID-19 epidemic, there was a significant decrease in number of inpatients(280±157.1 vs 340.4±110.2), outpatient(including emergency)(8 359±3 615 vs 10 151±3 225), the workload of blood typing(272.0±132.4 vs 341.6±110.4), unexpected antibody screening(78.26±42.22 vs 98.51±43.53) and crossmatch(237.2±99 vs 475.7±155.6), as well as the consumption(U) of all blood components(457.9±50.32 vs 579.4±62.51) in the Xijing Hospital(P<0.05). In detail, the epidemic had the most direct impact on the number of inpatients and outpatients, which shrank continuously on the 2^nd day after official announcement of the new COVID-1 cases. While the workload of blood typing, unexpected antibody screening and crossmatch decreased slightly, with a lag, usually on the 2^nd, 3^rd and 5^th day after official announcement. The decrease of the usage of red blood cells and plasma began from the 7^th day after the new epidemic to the 6^th day after the end of the epidemic. However, the usage platelets and cryoprecipitate coagulation factors decreased from the 8^th and 10^th day after the new epidemic to the 2^nd and 6^th day after the end of the epidemic, respectively. 【Conclusion】 The daily work of Blood Transfusion Department has been seriously affected by sporadic COVID-19 epidemic. The working mode, staff structure and inventory ratio of blood components should be adjusted and optimized instantly to maintain the normal conduct of medical treatments in hospitals and ensure the safety of patients.