OBJECTIVE: To observe the therapeutic effects and underlying mechanism of baicalin against colon cancer. METHODS: The effects of baicalin on the proliferation and growth of colon cancer cells MC38 and CT26. WT were observed and predicted potential molecular targets of baicalin for colon cancer therapy were studied by network pharmacology. Furthermore, molecular docking and drug affinity responsive target stability (DARTS) analysis were performed to confirm the interaction between potential targets and baicalin. Finally, the mechanisms predicted by in silico analyses were experimentally verified in-vitro and in-vivo. RESULTS: Baicalin significantly inhibited proliferation, invasion, migration, and induced apoptosis in MC38 and CT26 cells (all P<0.01). Additionally, baicalin caused cell cycle arrest at the S phase, while the G₀/G₁ phase was detected in the tiny portion of the cells. Subsequent network pharmacology analysis identified 6 therapeutic targets associated with baicalin, which potentially affect various pathways including 39 biological processes and 99 signaling pathways. In addition, molecular docking and DARTS predicted the potential binding of baicalin with cyclin dependent kinase inhibitor 2A (CDKN2A), protein kinase B (AKT), caspase 3, and mitogen-activated protein kinase (MAPK). In vitro, the expressions of CDKN2A, MAPK, and p-AKT were suppressed by baicalin in MC38 and CT26 cells. In vivo, baicalin significantly reduced the tumor size and weight (all P<0.01) in the colon cancer mouse model via inactivating p-AKT, CDKN2A, cyclin dependent kinase 4, cyclin dependent kinase 2, interleukin-1, tumor necrosis factor α, and activating caspase 3 and mouse double minute 2 homolog signaling (all P<0.05). CONCLUSION Baicalin suppressed the CDKN2A protein level to prevent colon cancer and could be used as a therapeutic target for colon cancer.
Flavonoids/pharmacology* ; Colonic Neoplasms/prevention & control* ; Animals ; Cell Line, Tumor ; Molecular Docking Simulation ; Cell Proliferation/drug effects* ; Apoptosis/drug effects* ; Cyclin-Dependent Kinase Inhibitor p16/metabolism* ; Mice ; Mice, Inbred BALB C ; Cell Movement/drug effects* ; Humans ; Gene Expression Regulation, Neoplastic/drug effects* ; Cell Cycle Checkpoints/drug effects*

To explore the effect and mechanisms of demethylation drug zebularine on esophageal cancer cells apoptosis, ECA109 cells and KYSE170 cells were treated with zebularine at different concentrations (25, 50, 100, 200, and 400 μmol·L^-1). The cell viability was measured by CCK-8. Flow cytometry was used to detect the cell apoptosis rate, Western blot was performed to determine the expression of apoptosis protein (Bcl-2, Bax, cleaved-caspase-3, and cleaved-PARP) and Wnt signal pathway molecules (β-catenin, cyclin D1, and c-Myc), real-time quantitative PCR was used to detect the expression level of negative regulatory genes of Wnt signaling pathway, methylation specific PCR (MSP) was used to detect the methylation status of secreted frizzled related protein 2 (SFRP2) and dickkopf 3 (Dkk3) genes. After knockdown of SFRP2 and Dkk3, the effect of zebularine on apoptosis was detected. The studies showed that zebularine could inhibit the activity of ECA109 and KYSE170 cells in a dose-dependent and time-dependent manner; zebularine could induce cell apoptosis, down-regulate the expression of Bcl-2 protein, up-regulate the expression of Bax, cleaved-caspase-3, and cleaved-PARP protein, and inhibit the expression of β-catenin, cyclin D1, and c-Myc protein (P < 0.05); the mRNA expression levels of Dkk3 and SFRP2 were significantly up-regulated by zebularine, while the methylation levels of SFRP2 and Dkk3 promoters were decreased; knockdown of SFRP2 and Dkk3 could reduce the apoptosis induced by zebularine. In summary, zebularine could reduce the methylation level of SFRP2 and Dkk3 gene promoter, promote the expression of SFRP2 and Dkk3 gene, and then induce the apoptosis of esophageal cancer cells by inhibiting Wnt/β-catenin signaling pathway.

Objective To investigate the prevalence and risk factors of Blastocystis hominis infections among patients with HIV/AIDS in Fuyang City, Anhui Province. Methods A cross-sectional study was conducted in Fuyang City, Anhui Province in 2016. The demographic and socioeconomic status, and the lifestyle and production style were collected using a questionnaire survey. B. hominis DNA was detected in subjects’stool samples using a PCR assay, and the CD4+ T lymphocyte count and HIV viral load were measured in the subjects’ blood samples. The risk factors of B. hominis infections among patients with HIV/AIDS were identified using univariate and multivariate logistic regression analyses. Results A total of 398 HIV/AIDS patients were enrolled in this study, with a mean age of 49.3 years, a mean body weight of 55.9 kg and a mean height of 164.4 cm. The prevalence of B. hominis infection was 6.78% in the study subjects, and no gender- (χ² = 1.589, P = 0.207), education level- (χ² =0.508, P = 0.776), marital status- (χ² = 0.419, P = 0.811) or occupation-specific prevalence (χ² = 2.744, P = 0.615) was detected. Among the patients with HIV/AIDS, there were no significant differences in the age (t = 0.370, P = 0.712), height (t = 1.587, P =0.113), body weight (t = 0.516, P = 0.606), CD4⁺ T lymphocyte count (t = 1.187, P = 0.230) or HIV viral load (t = 0.193, P =0.496) between B. hominis-infected and uninfected individuals. Dinking non-tap water [OR = 6.554, 95% CI: (1.876 to 22.903)] and keeping dogs [OR = 5.895, 95% CI: (2.017 to 17.225)] were identified as risk factors for B. hominis infection in patients with HIV/AIDS. Conclusion The prevalence of B. hominis infection is high in HIV/AIDS patients, and drinking non-tap water and keeping dogs are risk factors for B. hominis infection among HIV/AIDS patients.

Objective To explore the effect of different family tobacco control patterns on cotinine level and acute respiratory infections among infants.Methods A total of 300 infants were included,and were divided into 3 groups based on the tobacco control patterns:strictly tobacco -controlled group (97 cases),partly tobacco -controlled group (88 cases)and tobacco -uncontrolled group (1 1 5 cases).Urinary cotinine was measured in all participants.All participants were prospectively followed -up for 1 year,and the incidence of acute respiratory infections was recorded during the follow-up.Results The cotinine level of strictly tobacco -controlled group [0.45 ±0.21 (μg/L)]was significantly lower than the other two groups [1 .01 ±0.49(μg/L),1 .1 6 ±0.48(μg/L),P <0.05],and no significant differences were detected between the partly tobacco -controlled group and tobacco -uncontrolled group.The incidence of lower respiratory tract infection,not the upper respiratory tract infections,was significant different among the 3 groups (strictly tobacco -controlled group:1 8.75%;partly tobacco -controlled group:32.1 8%;tobacco -uncontrolled group:37.72%)(P <0.05).The number of upper respiratory tract infections and lower respiratory tract infections was significantly different among the three groups.The difference was significant between strictly tobacco -controlled group and partly tobacco -controlled group and between strictly tobacco -controlled group and tobacco -uncontrolled group (P <0.05).Conclusion Strict tobacco control could reduce the prevalence of passive smoking and the incidence of respiratory infections among infants.

OBJECTIVETo investigate the effect of interferon regulatory factors (IRFs) on neointimal formation after vascular injury in the mouse, and its possible mechanism.

METHODSVascular injury was induced by polyethylene cuff placement around the left femoral artery of IRF-1-deficient mice and C57BL/6J mice. The mRNA expressions of IRF-1, IRF-2, angiotensin II type 2 (AT2) receptor, interleukin-1 beta converting enzyme (ICE), inducible nitric oxide synthase (iNOS) were detected by RT-PCR and immunohistochemical staining.

RESULTSNeointimal formation after vascular injury was significantly greater in IRF-1-deficient mice than that in C57BL/6J mice (P<0.05). In contrast, TUNEL-positive nuclei to total nuclei in the neointima and media in vascular smooth muscle cell (VSMC) in the injured artery significantly attenuated in IRF-1-deficient mice compared to C57BL/6J mice (P<0.05). The expressions of AT2 receptor as well as pro-apoptotic genes such as ICE and iNOS in C57BL/6J mice were up-regulated in response to vascular injury, but this upregulation was attenuated in IRF-1-deficient mice.

CONCLUSIONSOur results suggest that IRF-1 induces VSMC apoptosis and inhibits neointimal formation after vascular injury at least partly due to the upregulation of AT2 receptor, ICE and iNOS expressions. These results indicate that IRF-1 exerts an inhibitory effect on neointimal formation through the induction of apoptosis in VSMCs.

Animals ; Apoptosis ; physiology ; Caspase 1 ; genetics ; metabolism ; Femoral Artery ; anatomy & histology ; pathology ; Interferon Regulatory Factor-1 ; genetics ; metabolism ; Interferon Regulatory Factor-2 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Smooth, Vascular ; cytology ; metabolism ; pathology ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; metabolism ; Receptor, Angiotensin, Type 2 ; genetics ; metabolism ; Tunica Intima ; pathology ; physiology

OBJECTIVETo provide right time points in selection of right aged animals and the normal physiological data of TX mice.

METHODS7-12 months old TX and DL mice were studied, each group contained 3 female and 3 male mice of TX or DL mice. The concentration of copper in the serum, dry tissues (liver, brain and kidney), together with copper biochemistry indexes were measured. The liver histopathology was observed under light microscopy and electron microscope.

RESULTSTransaminase increased significantly only in 10 and 11-month- old (AST(TX10) = 218.3 U/L, AST(TX11) = 197.5 U/L, AST(DL10) = 171.5 U/L, AST(DL11) = 165.0 U/L, P(10) less than 0.001, P(11) = 0.022), but the copper concentration of liver, brain and kidney was significantly increased during 7-12 month old (the average concentration of copper, Liver(TX) = (750.0 +/- 85.5) mg/kg, Brain(TX) = (39.7 +/- 2.2)mg/kg, Kidney(TX) = (29.8 +/- 5.0) mg/kg, Liver(DL) = (11.6 +/- 1.5) mg/kg, Brain(DL) = (16.8 +/- 0.9) mg/kg, Kidney(DL) = (14.2 +/- 1.0) mg/kg, t = 21.16, 23.60, 7.47, for all these organs P less than 0.05).

CONCLUSIONTX mice is a suitable model of liver disease with natural recovery, so selecting animal model of suitable time point is very important.

Animals ; Aspartate Aminotransferases ; blood ; Brain ; metabolism ; Ceruloplasmin ; metabolism ; Copper ; metabolism ; Disease Models, Animal ; Female ; Kidney ; metabolism ; Liver ; metabolism ; pathology ; Liver Diseases ; blood ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Time Factors

Objective To observe the changes of brain heme oxygenase-1/carbon monoxide (HO-1/CO)in neonatal rats with hypoxic-ischemic brain damage(HIBD)injury and investigate the role of HO-1/CO in the recovery of HIBD. Methods Eighteen 7-day-old Wistar rats were randomly divided into sham-operated group,HIBDgroup and HIBDwith zinc protoporphyrin(ZnPP)treatment group (n=6).In the latter two groups,HIBD model was established by unilateral carotid ligation followed by timed exposure to 8%oxygen. Real-time fluorescent quantitative PCR Was performed to determine the expression of HO-1 mRNA and thiobarbituric acid(TBA)method was used to assay malondialdehyde (MDA)contentinthe braintissue of the rats.The cell apoptosis in the brain aRer HIBD was analyzed using flow cytometry,and the blood CO concentration was detected by the absorbance at 420mn and 432 nm.Results Compared to the sham-operated group.HO-1 mRNA expression and blood CO concentration were significantly increased in HIBD group and ZnPP group (P＜0.05).The rats with ZnPP group had significantly lower HO-1 mRNA expression and blood CO concentration than those in HIBD group(P＜0.05).HIBD resultedin significantly increased MDA content and cell apoptosis rate in the rat brain as compared to those in the sham-operated group(p＜0.05),and ZnPP treatment further increased the MDA content and cell apoptosis(P＜0.05). Conclusions Increased brain HO-1 mRNA expression and blood CO concentration in neonatal rats with HIBD are probably associated with the spontaneous recovery of neural tissue injury.