Objective Microcystin-leucine-arginine(MC-LR)exposure induces lipid metabolism disorders in the liver.Secreted frizzled-related protein 5(SFRP5)is a natural antagonist of winglesstype MMTV integration site family,member 5A(Wnt5a)and an anti-inflammatory adipocytokine.In this study,we aimed to investigate whether MC-LR can induce lipid metabolism disorders in hepatocytes and whether SFRP5,which has anti-inflammatory effects,can alleviate the effects of hepatic lipid metabolism by inhibiting the Wnt5a/Jun N-terminal kinase(JNK)pathway. Methods We exposed mice to MC-LR in vivo to induce liver lipid metabolism disorders.Subsequently,mouse hepatocytes that overexpressed SFRP5 or did not express SFRP5 were exposed to MC-LR,and the effects of SFRP5 overexpression on inflammation and Wnt5a/JNK activation by MC-LR were observed. Results MC-LR exposure induced liver lipid metabolism disorders in mice and significantly decreased SFRP5 mRNA and protein levels in a concentration-dependent manner.SFRP5 overexpression in AML12 cells suppressed MC-LR-induced inflammation.Overexpression of SFRP5 also inhibited Wnt5a and phosphorylation of JNK. Conclusion MC-LR can induce lipid metabolism disorders in mice,and SFRP5 can attenuate lipid metabolism disorders in the mouse liver by inhibiting Wnt5a/JNK signaling.

Owing to incurable castration-resistant prostate cancer (CRPC) ultimately developing after treating with androgen deprivation therapy (ADT), it is vital to devise new therapeutic strategies to treat CRPC. Treatments that target programmed cell death protein 1 (PD-1) and programmed death ligand-1 (PD-L1) have been approved for human cancers with clinical benefit. However, many patients, especially prostate cancer, fail to respond to anti-PD-1/PD-L1 treatment, so it is an urgent need to seek a support strategy for improving the traditional PD-1/PD-L1 targeting immunotherapy. In the present study, analyzing the data from our prostate cancer tissue microarray, we found that PD-L1 expression was positively correlated with the expression of heterogeneous nuclear ribonucleoprotein L (HnRNP L). Hence, we further investigated the potential role of HnRNP L on the PD-L1 expression, the sensitivity of cancer cells to T-cell killing and the synergistic effect with anti-PD-1 therapy in CRPC. Indeed, HnRNP L knockdown effectively decreased PD-L1 expression and recovered the sensitivity of cancer cells to T-cell killing in vitro and in vivo, on the contrary, HnRNP L overexpression led to the opposite effect in CRPC cells. In addition, consistent with the previous study, we revealed that ferroptosis played a critical role in T-cell-induced cancer cell death, and HnRNP L promoted the cancer immune escape partly through targeting YY1/PD-L1 axis and inhibiting ferroptosis in CRPC cells. Furthermore, HnRNP L knockdown enhanced antitumor immunity by recruiting infiltrating CD8⁺ T cells and synergized with anti-PD-1 therapy in CRPC tumors. This study provided biological evidence that HnRNP L knockdown might be a novel therapeutic agent in PD-L1/PD-1 blockade strategy that enhanced anti-tumor immune response in CRPC.

Objective: To understand the relationship between eye strain and eye health behavior in college students learning at home during the period of COVID-19 epidemic, and to provide a scientific reference for improving the hygiene of using eyes among the college students. Methods: A cross sectional study and stratified cluster sampling was used to select 2 671 college students from 8 colleges in Anhui Province during the March 1st to July 1st in 2020, and an online questionnaire was survey included general information,eye strain,and daily eye health behavior. Results: The prevalence of eye strain in college students was 69.64%. Multivariable Logistic regression analysis showed that eye strain was correlated with gender, myopia, siesta habit, staying up until 2:00 am, and the use of eye liquid, with OR values(95% CI ) were 0.64(0.53-0.76), 1.77(1.42- 2.20 ),0.71(0.59-0.86), 1.39(1.17-1.65), and 2.18(1.71-2.79), respectively. There was no correlation among daily outdoor activity time, daytime reading time and the occurrence of eye strain( P >0.05). Conclusion During the period of COVID-19 epidemic, eye strain among college students is common. The daily eye health behavior is related to the occurrence of eye strain. Under the special learning context, eye care measures should be encouraged specifically.

Objective To observe the effect of Huangxiong Kangshuan capsule on serum high-sensitivity C-reactive protein (hs-CRP) and interleukin-6 (IL-6) levels in patients with acute cerebral infarction. Methods Ninety-two patients with acute cerebral infarction admitted to the First Affiliated Hospital of Anhui University of Traditional Chinese Medicinefrom July 2013 to December 2016 were enrolled, and they were divided into an observation group (47 cases) and a control group (45 cases) by random number table. The control group was given conventional treatment of ischemic cerebrovascular disease, while the observation group was additionally treated by Huangxiong Kangshuan capsule orally taken, once 3 tablets, 3 times a day, on the basis of routine treatment; the duration of treatment was 2 weeks in both groups. After 2 weeks of treatment, the clinical effects of the two groups and the changes of serum hs-CRP and IL-6 levels were observed.Results After treatment, the levels of serum hs-CRP and IL-6 were decreased significantly compared with those before treatment in the two groups [observation group: hs-CRP (mg/L) was 6.18±2.17 vs. 14.11±3.01, IL-6 (ng/L): 28.10±11.47 vs. 120.83±24.51; control group: hs-CRP (mg/L) was 8.89±2.46 vs. 13.97±2.69, IL-6 (ng/L) was 49.48±16.43 vs. 115.25±24.05], and the degree of decline in the observation group was more significant than that in the control group [hs-CRP (mg/L): 6.18±2.17 vs. 8.89±2.46, IL-6 (ng/L): 28.10±11.47 vs. 49.48±16.43, bothP < 0.01]; the total effective rate of the observation group was significantly higher than that of the control group [87.2% (41/47) vs. 71.1% (32/45),P < 0.05]. Conclusion Huangxiong Kangshuan capsule can decrease the serum hs-CRP and IL-6 levels in patients with acute cerebral infarction, and has a role in brain protection and nerve function defect improvement.

Objective:To explore the clinical efficacy of alanyl glutamine (Ala-Gln) combined with octreotide in the treatment of severe acute pancreatitis (SAP) and its effects on the serum interleukin-6 (IL-6),C reactive protein (CRP) levels and related biochemical parameters.Methods:100 cases of patients with SAP in our hospital from January 2013 to December 2016 were selected and randomly divided into two groups.The control group was treated by octreotide,while the observation group was treated by Ala-Gln combined with octreotide.The clinical effect,changes of serum IL-6,CRP,amylase (AMY),lactate dehydrogenase (LDH),prealbumin (PA) and albumin (ALB) levels before and after therapy were compared between two groups.Results:On the 10th day after treatment,the overall effective rate of observation group was 88.0％ which was significantly higher than that of the control group (64.0％,P＜0.05).The serum IL-6,CRP,AMY and LDH levels of both groups on the 10th day after treatment were significantly lower than those before treatment (P＜0.01).Compared with the control group,the improvement of serum IL-6,CRP,AMY,LDH levels of observation group were more significant(P＜0.01).Compared with those before treatment,the serum PA and ALB levels of both groups were significantly increased on the 10th day after treatment(P＜0.05),and the improvement of serum PA and ALB levels of observation group on the 10th day after treatment were significantly better than those of the control group(P＜0.01).Conclusion:Alanyl glutamine combined with octreotide could effectively eliminate or alleviate the symptoms and signs of patients with severe acute pancreatitis,control the inflammatory reaction,improve the level of metabolism,improve the prognosis.

Objective To study changes in expression of claudin-11 and proteins related to mitogen-activated protein kinase (MAPK) signaling pathways, as well as the ultrastructure of the blood testis barrier(BTB), in male ICR mice exposed to decabromodiphenyl ether (BDE-209). Methods Fifty-two mice, 4 weeks of age, weighing 15-21 g, were provided with adaptive feeding for 1 week. Mice were randomly divided into 4 groups, named control, low-dose, medium-dose and high-dose groups. The treated groups received BDE-209, by intragastric gavage, at doses, respectively, of 100, 300 and 500 mg/kg. Mice were sacrificed after 6 weeks and organs harvested on ice, weighed and stored at-80 °C. The ultrastructure of testicular tissues was examined by electron microscopy. Western blotting was used to detect proteins related to the MAPK pathway, including p38 mitogen activated protein kinase (p38), phosphorylated p38 (p-p38), extracellular regulated protein kinase 1/2 (ERK1/2) , phosphorylated ERK1/2 (p-ERK1/2) , c-jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK) and the BTB tight junction protein claudin-11. Analyze the difference between each groups. Results At sacrifice, the body weights in each treated group were compared with those in the control group weighing (41.14 ± 0.60) g. Compared with controls, body weights were significantly different (P<0.05) in the middle dose, at (39.97 ± 0.66) g and high dose, at (39.98± 0.55) g in control group. The coefficients of the testis were significantly lower (P<0.05) in each treated group than in controls, with values of (0.37±0.0)%, (0.31±0.05)% and (0.31±0.04)% for low- dose, medium-dose and high-dose groups, respectively. The epidymus coefficient values were also significantly lower than controls (P<0.05), with values of (0.16±0.06)%, (0.11±0.05)% and (0.07±0.03)%, respectively in the same three dose groups. Electron microscopy ultrastructure showed that, compared with the control group, the testes in the middle and high dose groups had closely connected fractures, cell edema and more vacuoles. Compared with in the control group, levels of p-p38 and p-JNK in testicular tissue were significantly increased (P<0.05). In the control group and in low-, medium- and high-dose groups, the p-p38/p38 ratios were 1.35±0.13, 3.46±0.10, 5.71±0.26 and 4.79±0.21, respectively. The corresponding p-JNK/JNK ratios were 2.07±0.0, 4.77±0.18, 3.63±0.06 and 4.85±0.15. Claudin-11 levels were significantly lower (P<0.05) than control values in each dosed group. The corresponding values in control, low-dose, medium-dose and high-dose groups were 8.33±0.36, 2.06±0.27, 3.37±0.27 and 1.55±0.19, respectively. Conclusion BDE-209 increased expression of proteins in the MAPK pathway and decreased expression of the BTB tight junction protein claudin-11 in testicular tissue. It also caused ultrastructural damage to the Sertoli cell BTB tight junctions.This suggested that BDE-209 might damage Sertolicells BTB through effects on the MAPK pathway.

Objective To study changes in expression of claudin-11 and proteins related to mitogen-activated protein kinase (MAPK) signaling pathways, as well as the ultrastructure of the blood testis barrier(BTB), in male ICR mice exposed to decabromodiphenyl ether (BDE-209). Methods Fifty-two mice, 4 weeks of age, weighing 15-21 g, were provided with adaptive feeding for 1 week. Mice were randomly divided into 4 groups, named control, low-dose, medium-dose and high-dose groups. The treated groups received BDE-209, by intragastric gavage, at doses, respectively, of 100, 300 and 500 mg/kg. Mice were sacrificed after 6 weeks and organs harvested on ice, weighed and stored at-80 °C. The ultrastructure of testicular tissues was examined by electron microscopy. Western blotting was used to detect proteins related to the MAPK pathway, including p38 mitogen activated protein kinase (p38), phosphorylated p38 (p-p38), extracellular regulated protein kinase 1/2 (ERK1/2) , phosphorylated ERK1/2 (p-ERK1/2) , c-jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK) and the BTB tight junction protein claudin-11. Analyze the difference between each groups. Results At sacrifice, the body weights in each treated group were compared with those in the control group weighing (41.14 ± 0.60) g. Compared with controls, body weights were significantly different (P<0.05) in the middle dose, at (39.97 ± 0.66) g and high dose, at (39.98± 0.55) g in control group. The coefficients of the testis were significantly lower (P<0.05) in each treated group than in controls, with values of (0.37±0.0)%, (0.31±0.05)% and (0.31±0.04)% for low- dose, medium-dose and high-dose groups, respectively. The epidymus coefficient values were also significantly lower than controls (P<0.05), with values of (0.16±0.06)%, (0.11±0.05)% and (0.07±0.03)%, respectively in the same three dose groups. Electron microscopy ultrastructure showed that, compared with the control group, the testes in the middle and high dose groups had closely connected fractures, cell edema and more vacuoles. Compared with in the control group, levels of p-p38 and p-JNK in testicular tissue were significantly increased (P<0.05). In the control group and in low-, medium- and high-dose groups, the p-p38/p38 ratios were 1.35±0.13, 3.46±0.10, 5.71±0.26 and 4.79±0.21, respectively. The corresponding p-JNK/JNK ratios were 2.07±0.0, 4.77±0.18, 3.63±0.06 and 4.85±0.15. Claudin-11 levels were significantly lower (P<0.05) than control values in each dosed group. The corresponding values in control, low-dose, medium-dose and high-dose groups were 8.33±0.36, 2.06±0.27, 3.37±0.27 and 1.55±0.19, respectively. Conclusion BDE-209 increased expression of proteins in the MAPK pathway and decreased expression of the BTB tight junction protein claudin-11 in testicular tissue. It also caused ultrastructural damage to the Sertoli cell BTB tight junctions.This suggested that BDE-209 might damage Sertolicells BTB through effects on the MAPK pathway.

OBJECTIVE: To investigate the effect of gene silencing of Bmi-1 on proliferation regulation of CD44+ nasopharyngeal carcinoma cancer stem-like cells (CSC-LCs). METHOD: The sequence-specific short hairpin RNA lentivirus targeting at human Bmi-1 gene (LV-Bmi-1shRNA) was constructed and was used to infect CD44+ nasopharyngeal carcinoma cells which were sorted by flow cytometry. A lentiviral which included a random sequence was also designed to serve as a negative control. We employed fluorescence microscope and flow cytometry to detect infection efficiency; real-time PCR was used to detect Bmi-1 and its downstream gene while each protein expression level was confirmed by western blotting protocol; CCK-8 proliferation assay was applied to measure proliferation capacity; tumor spheroid assay was used to evaluate the self-renewal capacity. Colony formation assay was used to measure cell colony formation capability; flow cytometry analyzed cell cycle distribution. RESULT: The constructed LV-Bmi-1shRNA successfully infected into the CD44+ nasopharyngeal carcinoma cells. The infection efficiency could reach above 95%; LV-Bmi-lshRNA effectively inhibited Bmi-1 mRNA and protein expression, while the downstream gene p16INK4a and p14ARF mRNA as well as protein expression level were upregulated (P < 0.05). Notablely, the proliferation, colony formation, self-renewal capabilities of the experimental group decreased significantly (P < 0.05). In addition, the cell cycle arrested at the G0-G1 phase. CONCLUSION Gene silencing of Bmi-1 inhibited the proliferation, colony formation and self-renewal capabilities of the CD44+ nasopharyngeal carcinoma CSC-LCs, inhibited the cell cycle processes, which may mediate through Bmi1-p16INK4a/p14ARF-p53 pathway. Our experimental results indicated that Bmi-1 gene may play an important role in the maintenance of the stem cell-like characteristics of CD44+ nasopharyngeal carcinoma cells. Bmi-1 gene may be a potential new target for the treatment of nasopharyng al carcinoma in the future.
Carcinoma ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Gene Silencing ; Humans ; Hyaluronan Receptors ; metabolism ; Lentivirus ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplastic Stem Cells ; cytology ; Polycomb Repressive Complex 1 ; genetics ; RNA, Messenger ; RNA, Small Interfering ; Tumor Suppressor Protein p14ARF ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective To identify a rapid and efficient fungal genomic DNA extraction method for PCR amplification.Methods Genomic DNA was extracted from Penicillum marneffei,Rhizopus microsporus,Cryptococcus neoformans and Candida albicans by heating pyrolysis,microwave,repeated freezing and thawing,lysozyme digestion,overnight snail enzymatic and Qiagen kit methods.DNA electrophoretogram was observed by gel imaging system.The concentration and purity of extracted DNA were determined with an ultramicro nucleic acid protein tester and the yields were calculated.PCR amplification and sequencing were also performed.ANOVA and SNK-q test were used for data analysis.Results There were statistical differences in concentrations and yields of the fungal DNA extracted from Penicillum marneffei (hyphal phase and yeast phase),Rhizopus microsporus,Coptococcus neoformans and Candida albicans by six methods (F=750.83,220.95,669.35,132.01,510.20 and 1658.35,287.10,963.64,1147.77,4521.22,all P ＜0.01).Of six methods,microwave method gained the highest DNA concentration and yield,followed by heating pyrolysis method,while Qiagen kit method obtained the lowest concentration and yield.All DNA extracted by 6 kinds of methods were positive in PCR amplification.Conclusion All of the six methods can be used for fungal DNA extraction which is sufficient for PCR amplification,but microwave and heating pyrolysis methods are more easy and simple to perform.

Hepatic eosinophilic granuloma (HEG) is a rare benign liver disease,which belongs to histocytosis.Preoperative diagnosis of HEG was difficult because its clinical manifestation was not characteristic.In this article,the clinical data of 1 patient with HEG who was treated at the Jinhua Municipal Central Hospital of Zhejiang Province in September 2012 were retrospectively analyzed,and the diagnosis,differential diagnosis and treatment for HEG were investigated.