Effects of flurochloridone on apoptosis and Nrf2/HO-1 and NFκB signaling pathways in mouse testis and TM4 cells / 环境与职业医学

Background Flurochloridone (FLC) is toxic to male reproduction and can induce apoptosis of testicular tissue and supporting cells under oxidative stress. Of particular concern is whether nuclear factor-erythrocyte 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway and nuclear factor kappa B (NFκB) signaling pathway participate this process. Objective To observe apoptosis of testicular tissue and sertoli TM4 cells and alterations of Nrf2/HO-1 and NFκB signaling pathways in mice treated with FLC in vivo/in vitro. Methods (1) Animal experiment. Testis samples were harvested from male C57BL/6 mice after 28-day FLC (0, 3, 15, 75, and 375 mg·kg−1 per day) exposure via oral route. Malondialdehyde (MDA) and superoxide dismutase (SOD) in homogenate of testicular tissue were measured by colorimetry. Apoptosis of testicular tissue was evaluated by TUNEL staining. Expression and distribution of Nrf2 and NFκB were detected by immunohistochemistry. Protein expression levels of Nrf2, HO-1, NAD(P)H: quinone oxidoreductase 1 (NQO1), NFκB, inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ), and phosphorylated recombinant inhibitory subunit of nuclear factor kappa-B alpha (P-IκBα) in testicular tissue homogenate were determined by Western blotting. (2) Cell experiment. TM4 cell lines were treated with 40, 80, 120, 160, and 200 μmol·L−1 FLC for 6 h, and cell viability was detected by CCK-8. After 6 h exposure to 40, 80, and 160 μmol·L−1 FLC, the apoptosis rate was detected by flow cytometry, and the protein expression levels of Nrf2, HO-1, NQO1, NFκB, IKKβ, and IκBα were detected by Western blotting. Results (1) Animal experiment. Apoptosis occurred in the interstitial and basal parts of spermatogenic tubules in male C57BL/6 mice after 28 days of oral FLC exposure. Compared with the control group, the MDA level in testicular tissue of the 375 mg·kg−1 FLC-treated group was significantly increased (P<0.05), and the SOD activity was significantly decreased (P<0.05). After 375 mg·kg−1 FLC exposure, apoptosis occurred in the interstitial and basal parts of spermatogenic tubules. The results of immunohistochemistry showed the expression of Nrf2 and NFκB in the interstitium and basal part of spermatogenic tubules of the treated groups. Compared with the control group, the protein levels of Nrf2, NQO1, P-IκBα, NFκB, and IKKβ in the 15, 75, and 375 mg·kg-1 groups were significantly increased (P<0.001), and the HO-1 protein level was significantly increased in the 375 mg·kg−1 group (P<0.001). (2) Cell experiment. Compared with the control group, the TM4 cell viabilities in the 40, 80, 120, 160, and 200 μmol·L−1 FLC-treated groups significantly decreased (P<0.01). The apoptosis rates were significantly increased (P<0.05), and the apoptosis rates increased from 5.7% in the control group to 7.4%, 9.4%, and 11.7% in the 40, 80, and 160 μmol·L−1, respectively. The Nrf2 protein level in the 40 μmol·L−1 group was significantly increased (P<0.01), while the levels significantly decreased in the 80 and 160 μmol·L−1 groups (P<0.01). The HO-1 protein levels in the 40, 80, and 160 μmol·L−1 groups were significantly increased (P<0.01). The level of NQO1 protein in the 40 μmol·L−1 group was significantly increased (P<0.01). The NFκB protein levels were significantly increased in the 80 and 160 μmol·L−1 groups (P<0.001). The IκBα protein levels were significantly decreased in all treated groups (P<0.001). The IKKβ protein had no significant change. Conclusion FLC induces testicular tissue apoptosis, and the process affects Nrf2/HO-1 signaling pathway and NFκB signaling pathway. The in vitro study confirms that FLC could induce apoptosis of TM4 cells and activate Nrf2/HO-1 and NFκB signaling pathways.

Multimodal imaging features of acute macular neuroretinopathy associated with COVID-19 / 中华眼底病杂志

Objective:To observe the multimodal imaging features of acute macular neuroretinopathy (AMN) associated with COVID-19.Methods:A retrospective case study. A total of 52 eyes of 26 patients of AMN associated with COVID-19 diagnosed in Handan Eye Hospital from December 8 to 20, 2022 were included in the study. There were 8 males and 18 females, with the mean age of (33.8±8.1) years. All the patients were bilateral. The time from diagnosis of COVID-19 to the onset of vision loss was 3 to 6 days. All patients underwent the examinations of best corrected visual acuity (BCVA), fundus color photography, infrared fundus photography (IR), fundus autofluorescence (AF), and optical coherence tomography (OCT). Fluorescein fundus angiography (FFA) combined with indoxine green angiography (ICGA) were performed in 12 eyes, and visual field were performed in 18 eyes. Multimodal image features of the affected eye were retrospectively analyzed.Results:The BCVA of the affected eye was 0.25 to 1.0. Round or mottled grayish-white lesions in the macular area were seen in all affected eyes. IR examination showed irregular map-like weak reflex in macular region. AF examination showed speckled fluorescence enhancement in lesion related areas in 3 eyes. FFA combined with ICGA showed weak fluorescence in the macular region in 8 eyes. OCT examination showed patchy strong reflection in the outer plexus layer (OPL) and outer nuclear layer (ONL) of macular area in all affected eyes, and partial absence of outer membrane and ellipsoid band. The en-face OCT showed petal-like intense refleciton between OPL and ONL. Eighteen eyes underwent visual field, and 15 eyes had central dark spots.Conclusion:The characteristic manifestations of AMN associated with COVID-19 are speckled or round-like grayish-white lesions in the macular area, weak reflexes in IR, enhanced OPL and ONL reflexes in OCT, and petal-like intense refleciton changes between OPL and ONL can be seen in en-face OCT.

Flurochloridone-induced apoptosis via IRE1α-JNK signaling pathway in mice testicular cells and TM4 cells / 环境与职业医学

Background Flurochloridone (FLC) can induce apoptosis in Sertoli cells, but the specific mechanism remains unknown. Objective To investigate the testicular cell apoptosis in mice as well as apoptosis and activation of endoplasmic reticulum stress in TM4 cell line induced by FLC through in vivo and in vitro study designs respectively, and study the role of inosital-requiring enzyme 1α (IRE1α)-c-Jun N-terminal kinase (JNK) signaling pathway in the process of FLC-induced apoptosis in TM4 cells through intervention study design. Methods Testicular tissues were collected from male C57BL/6 mice which were treated with 3, 15, 75, and 375 mg·（kg·d）−1 FLC by oral perfusion for 28 d. Apoptosis was observed by TUNEL staining, and the levels of apoptosis-related proteins were detected by Western blotting, including B-cell lymphoma-2 (Bcl-2), Bcl-2 interacting mediator of cell death (Bim), and Bcl-2 associated X protein (Bax). In the in vitro study, TM4 cells were treated with different concentrations of FLC (40, 80, and 160 μmol·L−1) for 6 h, then apoptosis rate was detected by flow cytometry, and the levels of apoptosis-related proteins (Bcl-2, Bim, and Bax) and endoplasmic reticulum stress-related proteins [glucose regulated protein 78 (GRP78), phosphorylated-protein kinase R like endoplasmic reticulum kinase (p-PERK), activating transcription factor 6 (ATF6), phosphorylated-inosital-requiring enzyme 1α (p-IRE1α), and phosphorylated-JNK (p-JNK)] were measured by Western blotting. In the intervention study, TM4 cells were pretreated with IRE1α phosphorylation inhibitor 4μ8C and JNK phosphorylation inhibitor SP600125 for 6 h, then treated with 160 μmol·L−1 FLC for 6 h. The levels of apoptosis-related proteins and endoplasmic reticulum stress-related proteins were measured by Western blotting, and cell viability was detected by cell counting kit-8. Results After the male C57BL/6 mice orally exposed to FLC for 28 d, apoptosis occurred in the seminiferous tubule. The protein expression level of Bcl-2, apoptosis inhibitor, was decreased in the 75 and 375 mg·(kg·d)−1 groups (P<0.05), and the protein expression levels of Bim and Bax, apoptosis promoters, were increased in the 75 and 375 mg·(kg·d)−1 groups respectively (P<0.05). The percentages of apoptotic cells in the 0, 40, 80, and 160 μmol·L−1 FLC groups were 2.7%±0.2%, 4.8%±1.3%, 9.4%±0.3%, and 13.2%±0.2%, respectively, increased significantly compared with the control group (P<0.05). The protein expression level of Bcl-2 also was decreased in the 160 μmol·L−1 FLC group (P<0.05), while the levels of Bim and Bax were increased in both of the 80 and 160 μmol·L−1 groups (P<0.05). The expression levels of endoplasmic reticulum stress-related proteins (GRP78, p-PERK, ATF6, p-IRE1α, and p-JNK) were increased (P<0.05) or showed a rising trend in TM4 cells. Pre-treatment with 4μ8C (25 and 50 μmol·L−1) and SP600125 (10 and 20 μmol·L−1) significantly down-regulated the protein expression levels of GRP78, p-IRE1α, p-JNK, and Bax induced by FLC (P<0.05) or in a downward trend. Both of the inhibitors alleviated the decreased cell viability induced by FLC (P<0.05) or in alleviating fashion. Conclusion FLC could induce apoptosis in mice testis and TM4 cell apoptosis through activating endoplasmic reticulum stress and IRE1α-JNK signaling pathway.

Relationship Analysis of CYP3A4*18 Gene Polymorphism with Therapeutic Efficacy and ADR of EGFR-TKI in Advanced NSCLC Patients Receiving Primary Treatment / 中国药房

OBJECTIVE:To investigate the relationship of Cytochrome P450 (CYP)3A4* 18 gene polymorphism with therapeutic efficacy and ADR of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) in non-small cell lung cancer (NSCLC) patients receiving primary treatment.METHODS:A total of 46 advanced NSCLC patients receiving primary EGFR-TKI (gefitinib or edotinib) treatment until disease progression or intolerance were selected from our hospital during Jan.2013-Mar.2016,and (gefitinib of erlotinib) treatment until disease progression or intolerance.CYP3A4*18 genotype was detected by PCR and direct sequencing.Clinical efficacies,progression-free survival (PFS) and the occurrence of ADR were compared among differ ent genotypes.RESULTS:Among 46 patients,there were 17 cases of CYP3A4*18 wild-type and 29 cases of CYP3A4*18 mutation-type,with mutation frequency of 63.0％.The objective response rate (ORR) of CYP3A4*18 wild-type patients was 23.5％,and disease control rate (DCR) of them was 70.6％.For CYP3A4*18 mutation-type patients,ORR and DCR were 27.6％ and 69.0％.There was no statistical significance in the proportion of patients with partial response,stable disease or progressive dis ease,ORR or DCR among different genotypes (P＞0.05).PFS of female patients were significantly longer than male patients;those of patients without smoking history were significantly longer than those with smoking history,with statistical significance (P＜0.05).There was no correlation between patients' age,therapy drugs,Eastern Oncology Collaboration scores,EGFR mutation types,CYP3A4*18 genotypes and PFS (P＞0.05).Patients'gender and smoking history were independent prognostic factors for PFS [odds ratios were 3.438,0.205,95％ CI were(1.393,8.488),(0.088,0.481)].Among CYP3A4* 18 wild-type patients,6 patients suffered from rash (35.3％) and 3 diarrhea (17.6％).Among mutation-type patients,26 patients suffered from rash (89.7％) and 15 diarrhea (51.7％),with statistical significance (P＜0.05).There was no statistical significance in the incidence of liver function injury and interstitial dermatitis among different genotypes (P＞0.05).CONCLUSIONS:CYP3A4*18 gene polymorphism may be not associated with therapeutic efficacy of EGFR-TKI in advanced NSCLC patients receiving primary treatment,but it is correlated with the occurrence of ADR.Mutation type patients are more likely to suffered from rash,diarrhea and other ADR.

Study on the Correlation between RRM1 Single Nucleotide Polymorphisms and Chemotherapy Sensitivity of NSCLC Patients for Gemcitabine / 中国药房

OBJECTIVE:To investigate the correlation between ribonucleotide reductase M1 subunit (RRM1) single nucleo-tide polymorphisms (SNPs) and chemotherapy sensitivity of patients with non-small cell lung cancer (NSCLC) for gemcitabine. METHODS:A total of 96 NSCLC patients receiving primary treatment selected from our hospital during Aug. 2014-Jul. 2016 were all accepted gemcitabine-based two-drug chemotherapy plan,with continuous treatment for at least 2 cycles(28 d as a cycle). Che-motherapy sensitivity rate was calculated by using the ratio of the sum of patients with complete response and partial response to the sum of test patients. RRM1 genotype was tested by PCR and direct sequencing. The correlation between different genotypes and chemotherapy sensitivity was analyzed. RESULTS:Distribution frequency of RRM1-37C>A CC, CA, AA genotype were 35.42％,52.08％,12.50％,respectively;distribution frequency of-524C>T CC,CT,TT genotype were 18.75％,37.50％, 43.75％,respectively. The frequency of each genotype was in the line with Hardy-Weinberg equilibrium(P>0.05). Chemotherapy sensitivity rate of 96 NSCLC patients was 37.50％. The patient's age,sex,ethnicity,smoking or not,TNM stage,pathological type,chemotherapy plan,and the Eastern American Oncology Collaboration score were not associated with chemotherapy sensitivi-ty (P>0.05). Chemotherapy sensitivity rates of RRM1(-37CA)+(-524CT)genotype and (-37CC)+(-524TT) genotype patients (57.14％,39.39％) were significantly higher than those of other genotype patients (10.71％),with statistical significance (P<0.05). There was no statistical significance in chemotherapy sensitivity rate between RRM1(-37CA)+(-524CT) and (-37CC)+(-524TT)genotype patients. CONCLUSIONS:In NSCLC patients,the SNPs of RRM1 can be used as predictive factor for the sen-sitivity of gemcitabine chemotherapy,and RRM1(-37CA)+(-524CT)and(-37CC)+(-524TT)genotype patients have higher sensi-tivity to this type of chemotherapy.

Relationship between pain severity, emotion and beliefs / 中国实用护理杂志

Objective To analyze the relationship between pain sensation, emotion and recognition in three dimensions. Methods By using questionnaires which contained general information questionnaire, Cancer Pain Questionnaire, Self-reporting Inventory (SCL-90), Pain Beliefs and Perceptions Inventory (PBPI) to investigate pain sensation, emotion and recognition of 46 patients with cancer pain. Results There were 13(28.3%) cases sufferd from mild pain,17 (37.0%) cases were moderate pain, 16 (34.8% )cases were severe pain.As to the result of SCL- 90,patients showed obvious symptom in somatization, depression, anxiety and hostility.They holded deep belief of that pain was very mysterious. There was a significant correlation between pain severity and depression(rs=0.377) , anxiety(rs=0.388) on the condition that confidence level was 0.01;there was also a significant correlation between pain degree and interpersonal sensitivity(rs=0.308), hostility(rs=0.320) on the condition that confidence level was 0.05. As to pain beliefs, pain degree had a significant correlation with it in the dimension of pain as mystery (rs=0.529) and pain was persistent(rs=0.680) on the condition that confidence level was 0.01. Conclusions The survey shows a positive correlation between pain severity,emotion of pain(such as anxiety,depression, hostility and interpersonal sensitivity)and beliefs about pain as mystery or permanent.

To discuss the relationship between part of risk factors and the characters of coronary artery lesion of different gender patients with coronary heart disease / 中国医师进修杂志

Objective To discuss the relationship between part of risk factors and the characters of coronary artery lesion of different gender patients with coronary heart disease(CHD). Methods Two hundred and eight patients who were diagnosed CHD by coronary angiography (CAG) were selected. All patients were divided into male group (122 cases) and female group (86 cases). The characters of CHD and part of clinical data in different gender CHD patients were compared and analyzed. The independent risk factors of different gender CHD patients were analyzed. Results The age of onset in female group was later than male group:(69.22±10.12) years vs.(62.80±11.34) years, P=0.000. The incidence rate of hypertension and diabetes in female group were significantly higher than those in male group: 83.7%(72/86) vs. 63.1%(77/122), 53.5%(46/86) vs. 32.8%(40/122);and the levels of total cholesterol(TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) in female group were significantly higher than those in male group:(5.16±1.26) mmol/L vs. (4.60±1.23) mmol/L,(1.16±0.27) mmol/L vs. (1.05±0.27) mmol/L, (3.17±1.16) mmol/L vs. (2.74±1.06) mmol/L;the level of uric acid (UA) in female group was significantly lower than that in male group: (319.83±90.05)μmol/L vs. (357.91±98.51)μmol/L, there were significantly differences(P<0.01). The level of trigalloyl glycerol (TG) in two groups had no significant difference: (1.91 ± 1.23) mmol/L vs. (1.75±0.97) mmol/L, P=0.298. Logistic regression analysis showed that age and diabetes were the risk factors of CHD in different gender (P<0.01 or <0.05). Conclusions There are different risk factors between different gender CHD patients, while the coronary artery lesion is similar. Diabetes is the most important independent risk factor of different gender CHD patients, which is more important for female patients with more risk factors.

Measures of improving innovation quality of medical students / 中华医学教育探索杂志

Improving the innovation quality of medical students is the key for cultivating the medical staff with international competition. The most important point for improving the innovation quality is to establish an educational model in favor of cultivation of medical students.Adjusting curriculum system,conforming experimental content and enhancing extracurricular research activity can make for the improvement of medical stndents' innovation and cultivation of scientific research quality.