Licochalcone A Induces Ferroptosis in Hepatocellular Carcinoma via Reactive Oxygen Species Activated by the SLC7A11/GPX4 Pathway.

Liver cancer is a common malignant tumor, and its incidence is increasing yearly. Millions of people suffer from liver cancer annually, which has a serious impact on global public health security. Licochalcone A (Lico A), an important component of the traditional Chinese herb licorice, is a natural small molecule drug with multiple pharmacological activities. In this study, we evaluated the inhibitory effects of Lico A on hepatocellular carcinoma cell lines (HepG2 and Huh-7), and explored the inhibitory mechanism of Lico A on hepatocellular carcinoma. First, we evaluated the inhibitory effects of Lico A on hepatocellular carcinoma, and showed that Lico A significantly inhibited and killed HepG2 and Huh-7 cells in vivo and in vitro. Transcriptomic analysis showed that Lico A inhibited the expression of solute carrier family 7 member 11 (SLC7A11), which induced ferroptosis. We confirmed through in vivo and in vitro experiments that Lico A promoted ferroptosis in hepatocellular carcinoma cells by downregulating SLC7A11 expression, thereby inhibiting the glutathione (GSH)-glutathione peroxidase 4 (GPX4) pathway and inducing activation of reactive oxygen species (ROS). In this study, we suggest that Lico A is a potential SLC7A11 inhibitor that induces ferroptotic death in hepatocellular carcinoma cells, thereby providing a theoretical basis for the development of natural small molecule drugs against hepatocellular carcinoma.

A bZIP transcription factor (CiFD) regulates drought- and low-temperature-induced flowering by alternative splicing in citrus.

Drought and low temperature are two key environmental factors that induce adult citrus flowering. However, the underlying regulation mechanism is poorly understood. The bZIP transcription factor FD is a key component of the florigen activation complex (FAC) which is composed of FLOWERING LOCUS T (FT), FD, and 14-3-3 proteins. In this study, isolation and characterization of CiFD in citrus found that there was alternative splicing (AS) of CiFD, forming two different proteins (CiFDα and CiFDß). Further investigation found that their expression patterns were similar in different tissues of citrus, but the subcellular localization and transcriptional activity were different. Overexpression of the CiFD DNA sequence (CiFD-DNA), CiFDα, or CiFDß in tobacco and citrus showed early flowering, and CiFD-DNA transgenic plants were the earliest, followed by CiFDß and CiFDα. Interestingly, CiFDα and CiFDß were induced by low temperature and drought, respectively. Further analysis showed that CiFDα can form a FAC complex with CiFT, Ci14-3-3, and then bind to the citrus APETALA1 (CiAP1) promoter and promote its expression. However, CiFDß can directly bind to the CiAP1 promoter independently of CiFT and Ci14-3-3. These results showed that CiFDß can form a more direct and simplified pathway that is independent of the FAC complex to regulate drought-induced flowering through AS. In addition, a bHLH transcription factor (CibHLH96) binds to CiFD promoter and promotes the expression of CiFD under drought condition. Transgenic analysis found that CibHLH96 can promote flowering in transgenic tobacco. These results suggest that CiFD is involved in drought- and low-temperature-induced citrus flowering through different regulatory patterns.

Nucleoside diphosphate-linked moiety X-type motif 15 R139C genotypes impact 6-thioguanine nucleotide cut-off levels to predict thiopurine-induced leukopenia in Crohn's disease patients.

BACKGROUND: Thiopurine-induced leukopenia (TIL) is a life-threatening toxicity and occurs with a high frequency in the Asian population. Although nucleoside diphosphate-linked moiety X-type motif 15 (NUDT15) variants significantly improve the predictive sensitivity of TIL, more than 50% of cases of this toxicity cannot be predicted by this mutation. The potential use of the 6-thioguanine nucleotide (6TGN) level to predict TIL has been explored, but no decisive conclusion has been reached. Can we increase the predictive sensitivity based on 6TGN by subgrouping patients according to their NUDT15 R139C genotypes? AIM: To determine the 6TGN cut-off levels after dividing patients into subgroups according to their NUDT15 R139C genotypes. METHODS: Patients' clinical and epidemiological characteristics were collected from medical records from July 2014 to February 2017. NUDT15 R139C, thiopurine S-methyltransferase, and 6TGN concentrations were measured. RESULTS: A total of 411 Crohn's disease patients were included. TIL was observed in 72 individuals with a median 6TGN level of 323.4 pmol/8 × 108 red blood cells (RBC), which was not different from that of patients without TIL (P = 0.071). Then, we compared the 6TGN levels based on NUDT15 R139C. For CC (n = 342) and CT (n = 65) genotypes, the median 6TGN level in patients with TIL was significantly higher than that in patients without (474.8 vs 306.0 pmol/8 × 108 RBC, P = 9.4 × 10-5; 291.7 vs 217.6 pmol/8 × 108 RBC, P = 0.039, respectively). The four TT carriers developed TIL, with a median 6TGN concentration of 135.8 pmol/8 × 108 RBC. The 6TGN cut-off levels were 411.5 and 319.2 pmol/8 × 108 RBC for the CC and CT groups, respectively. CONCLUSION: The predictive sensitivity of TIL based on 6TGN is dramatically increased after subgrouping according to NUDT15 R139C genotypes. Applying 6TGN cut-off levels to adjust thiopurine therapies based on NUDT15 is strongly recommended.

Efficiency of Zoledronic Acid in Inhibiting Accelerated Periprosthetic Bone Loss After Cementless Total Hip Arthroplasty in Osteoporotic Patients: A Prospective, Cohort Study.

OBJECTIVE: To investigate the influence of preoperative osteopenia/osteoporosis on periprosthetic bone loss after total hip arthroplasty (THA) and the efficiency of zoledronate (ZOL) treatment in periprosthetic bone preservation. METHODS: This multicenter, prospective cohort study was conducted in four centers between April 2015 and October 2017. Patients were assigned to Normal BMD, Osteopenia, and Osteoporosis+ZOL groups. Patients with osteopenia received daily oral calcium (600 mg/d) and vitamin D (0.5 µg/d), while patients in the Osteoporosis+ZOL group received additional ZOL annually (5 mg/year). Periprosthetic bone mineral density (BMD) in seven Gruen zones, radiographic parameters, Harris hip score, EuroQol 5-Dimensions (EQ-5D) score, and BMD in hip and spine were measured within 7 days, 3 months, 12 months postoperation and annually thereafter. RESULTS: A total of 266 patients were enrolled, while 81 patients that completed the first year follow-up were involved in the statistical analysis. The mean follow-up time was 1.3 years. There were significant decreases of mean BMD in total Gruen zones (-4.55%, P < 0.05) and Gruen zone 1 (-10.22%, P < 0.01) in patients with osteopenia during the first postoperative year. Patients in the Osteoporosis+ZOL group experienced a marked increase in BMD in Gruen zone 1 (+16%) at the first postoperative year, which had a significant difference when compared with the Normal BMD group (P < 0.05) and the Osteopenia Group (P < 0.001). Low preoperative BMD in hip and spine was predictive of bone loss in Gruen zone 1 at 12 months after THA in patients with normal BMD (R2 = 0.40, P < 0.05). CONCLUSIONS: Patients with osteopenia are prone to higher bone loss in the proximal femur after cementless total hip arthroplasty (THA). ZOL, not solely calcium and vitamin D, could prevent the accelerated periprosthetic bone loss after THA in patients with osteopenia and osteoporosis.

Receptor for advanced glycation end products involved in lung ischemia reperfusion injury in cardiopulmonary bypass attenuated by controlled oxygen reperfusion in a canine model.

Controlled oxygen reperfusion could protect the lung against ischemia-reperfusion injury in cardiopulmonary bypass (CPB) by downregulating high mobility group box 1 (HMGB1), a high affinity receptor of HMGB1. This study investigated the effect of controlled oxygen reperfusion on receptor for advanced glycation end products (RAGE) expression and its downstream effects on lung ischemia-reperfusion injury. Fourteen canines received CPB with 60 minutes of aortic clamping and cardioplegic arrest followed by 90 minutes of reperfusion. Animals were randomized to receive 80% FiO2 during the entire procedure (control group) or to a test group receiving a controlled oxygen reperfusion protocol. Pathologic changes in lung tissues, RAGE expression, serum interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were evaluated. The lung pathologic scores after 25 and 90 minutes of reperfusion were significantly lower in the test group compared with the control group (p < 0.001). RAGE expression, TNF-α, and IL-6 were downregulated by controlled oxygen treatment (p < 0.001). RAGE might be involved in the lung ischemia-reperfusion injury in canine model of CPB, which was downregulated by controlled oxygen reperfusion.

Controlled oxygen reperfusion protects the lung against early ischemia-reperfusion injury in cardiopulmonary bypasses by downregulating high mobility group box 1.

Restricting oxygen delivery during the reperfusion phase of cardiopulmonary bypass (CPB) protects the heart, but effects on lung ischemia reperfusion (IR) in CPB are unknown. We examined whether extracellular high mobility group box 1 (HMGB1) mediated inflammation during early lung IR injury in CPB. Fourteen healthy canines received CPB with 60 minutes of aortic clamping and cardioplegic arrest, followed by 90 minutes reperfusion. Following surgery, the animals were randomized into control (n = 7) or test (n = 7) groups. Control animals received a constant level of 80% FiO(2) during the entire procedure, and the test group received a gradual increase in FiO(2) during the first 25 minutes of reperfusion. In the test group, the FiO(2) was initiated at 40% and increased by 10% every 5 minutes, to 80%. Histology, lung injury variables, HMGB1 expression, and inflammatory responses were assessed at baseline (T1) and at 25 minutes (T2) and 90 minutes (T3) after starting reperfusion. Treatment with controlled oxygen significantly suppressed lung pathologies, lung injury variables, and inflammatory responses (all P < .001). After lung IR injury, HMGB1 mRNA and protein expressions were significantly decreased in the controlled oxygen group (all P < .001). Controlled oxygen reperfusion is protective in the early stages of lung IR injury in a canine CPB model, and this protection is linked to HMGB1 downregulation.

[Relationship between metabolic syndrome and general habits in daily life].

OBJECTIVE: To study the relationship between life style and metabolic syndrome (MS). METHODS: Using identical protocol and questionnaire, 173 patients with MS and 173 without MS selected from the general population undergoing physical examinations in Guangdong General Hospital were surveyed for the risk factors of MS in their life style, and logistic regression analysis was performed to identify the risk factors. RESULTS: MS was significantly related to overweight, prolonged maintenance of sitting posture, and heavy life or work stresses, among which overweight was the most important factor contributing to MS (OR=11.442, P<0.000). Regular exercise, long exercise time, and anxiety were protective factors for MS. Meat intake, smoking, time of getting sleep and insomnia were not found to relate to MS. CONCLUSION: MS is correlated to some general habits in daily life, and a healthy life style may help in the prevention and treatment of MS.

Circadian variation of plasma cortisol and whole blood reduced glutathione levels in nasopharyngeal carcinoma patients.

BACKGROUND & OBJECTIVE: Glutathione is involved in cellular protection against radiation damage and drug detoxification. This study was to investigate the circadian variation of plasma cortisol and whole blood reduced glutathione (GSH) levels in nasopharyngeal carcinoma (NPC) patients to provide references for chronotherapy for NPC. METHODS: A total of 13 NPC patients and 14 healthy volunteers were involved. Peripheral venous blood was sampled every 4 h during one 24-hour period starting at 12:00 am. The plasma cortisol concentration was determined by radioimmunoassay; the GSH concentration was determined by high performance liquid chromatography. RESULTS: Plasma cortisol levels of both groups displayed clear and similar circadian rhythms. The cortisol level peaked in both groups in the morning and was the lowest at mid-night. In both groups, GSH concentrations showed significant differences according to sampling time (ANOVA for Repeated Measures, F=5.18, P=0.02). By cosinor analysis, the circadian variation of the GSH level in NPC group was marginally statistically significant (Cosinor analysis, P=0.06) with the acrophase appeared at 05:02; the GSH level in control group displayed an obvious circadian rhythm and the acrophase appeared at 07:44+/-01:56 (P<0.01). The rhythm-adjust mean value of glutathione was (19.60+/-1.11) nmol/mg protein in NPC group and (8.95+/-0.46) nmol/mg protein in control group. CONCLUSIONS: The circadian rhythm of the plasma cortisol level is maintained in NPC patients, even in some patients at advanced stages. In NPC patients, GSH shows a trend of circadian variation which is similar to that in healthy controls. These results could be used to select special schedules for chrono-radiotherapy and chrono-chemotherapy for NPC patients.

[Induction of chemical hepatocarcinogenesis and establishment of orthotopic liver transplantation model with hepatocarcinoma in rats].

OBJECTIVE: To explore the inducing method for hepatic carcinoma in rats and the operative technique in establishment of orthotopic liver transplantation (OLT) model in rats with induced hepatic tumor. METHODS: Hepatic carcinoma was induced by diethylnitrosamine (DENA) in rats. Then OLT was performed, using a two-cuff vessel anastomosis method modified from that introduced by Kamada, with reconstruction of hepatic artery. RESULTS: At the observation time point (18 weeks) after the initiation of carcinogenesis, the one-month survival rate in OLT group was higher than that in non-OLT group, 56.3% and 21.4% respectively, P < 0.05. The operative successful rate in tumor rats group and control group was 87.5% and 90.0%, respectively. In OLT rats with hepatic carcinoma group, metastasis mainly occurred in lung and abdomen, while only 2 cases of intrahepatic recurrence were observed. There was significant difference between the tumor rats group and control group in cum survival rate. CONCLUSION: DENA can wholesale induce stable rat model for hepatic carcinoma. OLT can markedly elevate survival rate in rats with liver cancer. In rats with induced hepatic tumor, OLT model, suggested by the author, was established with hepatic arterial reconstruction and it offers a satisfied method with more similar pattern of physiological setting and carcinoma development situation.

[Correlation of GST-pi and PCNA expression to prognosis of advanced maxillary sinus squamous cell carcinoma].

BACKGROUND & OBJECTIVE: Tumor markers which relate to cell proliferation and metabolism have seldom been studied in maxillary sinus cancer. This study was conducted to identify the correlation of glutathione S-transferase pi (GST-pi) and proliferating cell nuclear antigen (PCNA) expression to prognosis of advanced maxillary sinus squamous cell carcinoma (SCC). METHODS: The expression of GST-pi and PCNA in 54 specimens of maxillary sinus SCC, 29 specimens of benign maxillary tumor, and 20 specimens of normal nasal mucosa was detected by immunohistochemistry. The correlation of GST-pi and PCNA expression to prognosis of advanced maxillary sinus SCC was analyzed by Kaplan-Meier method. The prognosis was analyzed by Cox multivariate model. RESULTS: The overexpression rates of GST-pi and PCNA were significantly higher in maxillary sinus SCC than in benign maxillary tumor and normal nasal mucosa (74.1% vs. 89.6% and 15.0%, P<0.01; 79.6% vs. 3.4% and 0, P<0.01). The 5-year survival rate was significantly higher in advanced maxillary sinus SCC patients with high expression of GST-pi than in the patients with low expression of GST-pi (34.5% vs. 21.2%, P=0.025); the difference between the patients with high and low expression of PCNA was not significant (18.0% vs. 27.0%, P=0.890). The expression of GST-pi was an independent prognostic factor of advanced maxillary sinus SCC (P=0.039, odds ratio>1). CONCLUSION: The overexpression of GST-pi is an independent prognostic factor of advanced maxillary sinus SCC, but that of PCNA isn't.

[Prognostic analysis of 151 patients with maxillary sinus malignant neoplasms].

BACKGROUND & OBJECTIVE: Five-year survival rate of patients with maxillary malignant neoplasms is low, the prognostic factors of these neoplasms were unclear. This study was to investigate prognostic factors of maxillary sinus malignant neoplasms. METHODS: Records of 151 inpatients with malignant neoplasms of maxillary sinus initially treated at Cancer Center of Sun Yat-sen University from Sep. 1983 to Mar. 1999 were reviewed. Of 151 cases, 72 were squamous cell carcinoma (SCC), 44 were adenocarcinoma, 16 were sarcoma, and 19 were other histological types; according to 1997 UICC classification, 7 were stage II, 55 were stage III, and 89 were stage IV; 66 patients received combined therapy of surgery and radiotherapy, 14 received surgery alone, 25 received radiotherapy alone, 39 received other treatments, and 7 gave up treatment. All patients were followed up for more than 5 years. Influences of clinicopathologic factors on prognosis of patients with maxillary sinus malignant neoplasms were analyzed by Kaplan-Meier method, and Cox regression model with SPSS10.0 software. RESULTS: Five-year overall survival rate of patients of 40 years old was 33.3%(P=0.030); that of patients with SCC was 30.2%, of patients with adenocarcinoma was 57.5%, of patients with sarcoma was 24.3%, of patients with tumor of other histological types was 50.7% (P=0.011); that of patients with tumor of stage II, III, and IV were 85.7%, 45.8%, and 32.7%, respectively (P=0.029); that of patients with cervical metastases was 14.4%, of patients without cervical metastases was 44.1% (P=0.005); that of patients with distant metastases was 14.3%, of patients without distant metastases was 41.1% (P=0.011); that of patients without treatment was 14.3%, of patients treated with surgery alone was 42.9%, of patients treated with radiotherapy alone was 32.3%, of patients treated with combined therapy of surgery and radiotherapy was 50.8%, of patients treated with other treatments was 29.1% (P=0.004). Univariate survival analysis showed that the above 6 factors were prognostic factors of patients with maxillary sinus malignant neoplasms. Multivariate analysis showed that combination of surgery and radiotherapy (P=0.004, OR< 1), clinical stage (P=0.025, OR >1), SCC (P=0.016, OR >1), and sarcoma (P=0.003, OR >1) were independent prognostic factors of patients with maxillary sinus malignant neoplasms. CONCLUSION: For maxillary sinus malignant neoplasms, patients with SCC or sarcoma had poorer survival than patients with adenocarcinoma or other histological types of tumor; patients with sarcoma had poorer survival than patients with SCC. The higher the patient's clinical stage was, the worse his prognosis was. Combination of surgery and radiotherapy may be the best treatment for patients with maxillary sinus malignant neoplasms.