BACKGROUND:Previous studies have shown that N-methyl-D-aspartic acid receptor(NMDA)receptors are associated with fluorine,but the role in fluoride-induced endoplasmic reticulum stress remains unclear. OBJECTIVE:To observe the changes of excitatory neurotransmitter NMDA receptor and endoplasmic reticulum stress IRE1α-ASK1-JNK pathway protein expression in brain tissue of rats with experimental fluorosis,and to investigate the pathogenesis of neurological injury in fluorosis by giving NMDA receptor inhibitor to SH-SY5Y cells. METHODS:(1)Animal model:18 1-month-old SD rats were randomly divided into control group(drinking water fluoride content<0.5 mg/L),low fluoride group(drinking water fluoride content 10.0 mg/L)and high fluoride group(drinking water fluoride content 100.0 mg/L),with 6 rats in each group,half of each sex.After 6 months of fluoride intake,the rats were observed for the occurrence of dental fluorosis,and the 24-hour urinary fluoride content was measured.After anesthesia and euthanasia,the brain tissue of rats was taken to observe the pathological changes.Western blot assay was used to detect NMDA receptors and IRE1α,ASK1 and JNK protein expression in the brain tissue.(2)Cell model:SH-SY5Y cells were cultured in vitro and treated with sodium fluoride at final concentrations of 0.3 mmol/L and 3 mmol/L.The fluoride-stained cells were interfered with 10 μmol/L NMDA receptor antagonists Ifenprodil and MK-801 to observe the relevant protein changes. RESULTS AND CONCLUSION:(1)The incidence of dental fluorosis and urinary fluoride level in rats in the high fluoride group were significantly higher than that in the control and low fluoride groups(P<0.05).(2)Compared with the control group,the cytoplasm of neuronal cells in the CA3 area of the hippocampus in the low fluoride group was slightly more basophilic,while the neuronal cells in the CA3 area of the high fluoride group were disorganized,with increased basophilicity and some of the nuclei solidified.(3)In rat brain tissue,the expressions of NR2A in the high fluoride group and NR2B in the low fluoride group were significantly higher compared with the control group(P<0.05),and NR2B,IRE1,ASK1,and p-JNK protein expression levels were increased in the high fluoride group compared with the control and low fluoride groups(P<0.05).(4)In SH-SY5Y cells,NR1,NR2A and NR2B protein expressions were significantly increased in the high fluoride group compared to the control group(P<0.05).The protein levels of NR1 and NR2A were significantly reduced in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).NR2B protein expression was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(5)In SH-SY5Y cells,IRE1,ASK1,and p-JNK protein expression was significantly higher in the high fluoride group compared with the control group(P<0.05),while ASK1 and p-JNK protein expressions were significantly decreased in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).IRE1 protein level was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(6)It is concluded that excessive fluorine intake activates NMDA receptors in the central nervous system,causing increased expression of endoplasmic reticulum stress IRE1α,ASK1,and p-JNK proteins,and the use of NMDA receptor inhibitors has a mitigating effect on endoplasmic reticulum stress caused by fluorosis.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*

OBJECTIVE To investigate the pharmacological components and mechanism of Qingre huoxue decoction in improving myocardial ischemia-reperfusion injury（MIRI）. METHODS Forty-two rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， Qingre huoxue decoction low-dose， medium-dose and high-dose groups （4.94， 9.88， 19.79 g/kg），Qingre huoxue decoction drug-containing serum group （19.79 g/kg） and blank serum group （normal saline）， with 6 rats in each group. Each group was given corresponding drug/normal saline intragastrically， once a day， for consecutive 2 weeks. Twelve hours after last administration， except for serum groups， MIRI model was induced in other groups （only threading without ligation in sham operation group）. After modeling， cardiac histopathology was observed and apoptosis level was detected. UPLC-MS was used to analyze the samples in Qingre huoxue decoction drug-containing serum group and blank serum group. The main pharmacological components were screened with the help of relevant databases. Multivariate statistical methods were used to analyze the differential metabolites and related metabolic pathways. Validation test was performed based on oxidative stress indicators. RESULTS Qingre huoxue decoction could improve the pathological injury of cardiac tissue and decrease apoptosis rate of cardiac cells in MIRI model rats （P＜0.05）. Qingre huoxue decoction drug-containing serum contained 20 main pharmacological components such as baicalin， succinic acid， baicalein， cryptotanshinone， isoferulic acid， protocatechuic aldehyde. Qingre huoxue decoction could significantly up-regulate the levels of 15 metabolites including L-arginine， L-arginine， citric acid， glutathione， β-D- glucose and L-carnitine， and down-regulated the levels of 14 metabolites including arachidonic acid， 3-phosphate-d-glycerol phosphate， linoleic acid， docosahexaenoic acid， phosphatidylcholine and lysophosphatidylcholine （P＜0.05）. These metabolites were mainly involved in energy metabolism， inflammatory injury， oxidative stress and autophagy. Results of validation tests showed that Qingre huoxue decoction could significantly reduce the levels of malondialdehyde， and increased the levels of superoxide dismutase （except for low-dose group） and glutathione peroxidase significantly （P＜0.05）. CONCLUSIONS Qingre huoxue decoction can improve the injury of cardiac tissue in MIRI model rats. Its pharmacological components include baicalin， cryptotanshinone， isoferulic acid， protocatechualdehyde， etc. Furthermore， it may play a protective role in MIRI by improving myocardial energy metabolism， down-regulating oxidative stress， inhibiting inflammation infiltration.

Objective : To investigate the effects of one night of sleep deprivation on adults ’cognitive performance and sleepiness. Methods : The study employed a repeated⁃measures design. Participants performed cognitive performance tasks , which included the Continuous Performance Test and Digit Span Test and Karolinska Sleepiness Scale. The data were analyzed using one⁃way repeated⁃measures ANOVA. Results : 48 participants were included in this study. Sleep deprivation during night shift had a adversely affect on sustained attention performance and alertness (P < 0. 05) . Compared with the beginning of night shift work , the correct number decreased , the number of errors , missed number and reaction time increased , and the score of Karolinska sleepiness scale decreased after the night shift work. The sleep deprivation has no significant effect on working memory performance (P > 0. 05) . Conclusion One night of sleep deprivation has significant deleterious effects on cognitive performance and subjec⁃ tive sleepiness. However, no effect on the working memory performances has been found.

Objective:To investigate the efficacy and major adverse cardiovascular events(MACE)in elderly patients with ischemic cardiomyopathy-induced heart failure with reduced ejection fraction(HFrEF)treated with Sacubitril-valsartan at 3 years of follow-up.Methods:This was a single-center retrospective cohort study.The elderly patients with ischemic cardiomyopathy-induced HFrEF aged 60-85 years were diagnosed and treated in Beijing Anzhen Hospital from January 2018 to January 2020.A total of 120 continuously included elderly HFrEF patients treated with sacubitril valsartan were enrolled as the observation group, and 120 age-, gender-and B-type natriuretic peptide-matched elderly HFrEF patients treated with angiotensin converting enzyme inhibitor/angiotensin receptor blocker(ACEI/ARB)were enrolled as the control group.Structure and function of the heart were detected at 1-year follow-up.The MACE was observed, including cardiogenic death, readmission due to heart failure worsening and malignant arrhythmia at 3-year follow-up.Results:During follow-up(23.5±11.6)months, the visit of 18 cases was lost.There were no significant differences in the age, medical history, blood pressure between the two groups( P>0.05). Follow-up results showed that the improvement was better in observation group than in control group as follows: the left ventricular ejection fraction(45.8±9.4)% vs.(40.7±8.5)%, left ventricular end-diastolic diameter(56.5±8.3)mm vs.(59.2±7.3)mm, left ventricular end-systolic diameter(42.5±11.2)mm vs.(45.7±9.6)mm, left atrium inner diameter(49.1±8.7)mm vs.(51.2±7.7)mm, and left ventricular mass index(111.3±34.3)g/m 2vs.(119.7±31.5)g/m 2( t=4.41, 2.68, 2.38, 1.98 and 1.98, respectively, P<0.01 or 0.05). The rates of readmission due to heart failure worsening and the incidence of MACE were lower in the observation group than in the control group(21.7% or 26/120 vs.36.7% or 44/120, and 45.0% or 54/120 vs.71.7% or 86/120, χ2=6.54 and 17.55, P<0.05 or 0.01). In patients with the grade Ⅲ and Ⅳ New York Heart Association(NYHA)cardiac function, the incidence of MACE were lower in the observation group than in the control group(75.0% or 9/12 vs.100.0% or 14/14, χ2=5.10, P<0.05). Conclusions:Sacubitril-valsartan can improve cardiac structure and function, and decrease the incidence of MACE in elderly patients with HFrEF induced by ischemic cardimyopathy.

Objective:To investigate the disability status of the elderly in medical-nursing integrated pension institutions in Beijing and analyze its influencing factors.Methods:Using the convenient sampling method, a total of 359 elderly people from 5 medical-nursing integrated pension institutions in Beijing were selected from May to August 2019. General data and Elderly Disability Assessment Scale (EDAS) were used to evaluate the disability status of the included population, and univariate and multivariate analysis were conducted.Results:A total of 95.3% (342/359) of the elderly had disability. The single factor analysis result showed that male, low education level, a large number of chronic diseases, history of fall, history of smoking, history of drinking, widowed and elderly people living in nursing homes at their public expense were more disabled ( P<0.05) . Multivariate analysis showed that, history of falls, the number of chronic diseases greater than or equal to 3 and widowhood were the influencing factors for aggravated disability ( P<0.05) . Conclusions:In this survey, the rate of disability of the elderly in medical-nursing integrated pension institutions in Beijing is relatively high and the degree of disability is serious. The focus should be on the elderly who are male, with low educational level, smoking history, drinking history, falling history, comorbidities of the aged, the loss of spouse and those who bear the expenses of pension institutions at public expense.

Objective:To explore the curative effect of assisted surgery based on 3D printing technology on complex Pilon fractures.Methods:From June 2017 to June 2018, 52 patients with complex Pilon fracture treated in the General Hospital of Armed Police Navy were selected in the study.The patients were divided into study group ( n=26) and control group ( n=26) according to different operation mode.The control group was treated with traditional open reduction and internal fixation, and the study group was supplemented with 3D printing technology.The differences of treatment effect, operation time, intraoperative hemorrhage, fracture healing time, hospitalization time, incidence of complications, ankle function score and VAS pain score between the two groups were compared. Results:The total effective rate of the study group was 88.5%(23/26), which was higher than 65.4%(17/23) of the control group (χ 2=3.900, P=0.048). The mean operation time ( t=-3.770, P<0.001), mean fracture healing time ( t=-2.206, P=0.032) and mean hospitalization time ( t=-9.542, P<0.001) of the study group were shorter than those of the control group, and the amount of intraoperative bleeding ( t=-20.226, P<0.001) was less than that of the control group, the differences were statistically significant.The total incidence of complications in the study group was 15.4%(4/23), which was lower than 42.3%(11/23) in the control group (χ 2=4.591, P=0.032). With the development of time, the ankle function score of the two groups increased gradually, and the VAS pain score decreased gradually ( P<0.05). One month after operation, the ankle function score of the study group was higher than that of the control group ( t=2.340, P=0.023), but there was no statistically significant difference between the two groups in VAS pain score ( t=-0.278, P=0.782). At the last follow-up, the ankle function score of the study group was higher than that of the control group( t=2.760, P=0.008), and the VAS pain score of the study group was lower than that of the control group( t=-3.633, P<0.001), the differences were statistically significant. Conclusion:3D printing assisted surgery is better than traditional open reduction and internal fixation in the treatment of complex Pilon fracture, with shorter operation time, fracture healing time and hospitalization time, less intraoperative bleeding, lower incidence of complications and better postoperative recovery.

Objective:To study the patterns of tocilizumab (TCZ) use, its efficacy and safety in patients with rheumatoid arthritis (RA) in routine clinical practice.Methods:A total of 407 patients with RA were enrolled from 23 centers and treated with TCZ within 8 weeks prior to the enrollment visit, and were followed for 6-month. The patterns of TCZ treatment at 6 months, the effectiveness and safety outcomes were recorded. Statistical analysis was performed using SAS version 9.4.Results:A total of 396 patients were included for analysis, in which 330 (83.3%) patients received TCZ combined with conventional synthetic disease-modifying antirheumatic drugs (csDMARDs), and 16.7%(66/396) received TCZ monotherapy. At baseline, TCZ was initiated in 56.6%(224/396) and 9.6%(38/396) of patients after failure of DMARDs and other biological agents (bDMARDs) respectively. During the 6-month follow-up period, the mean frequency of TCZ administration was (3.7±1.6), the mean TCZ dosage was (7.4±1.2) mg/kg, and the mean interval between doses was (40±13) days. 120(25.8%) patients were on TCZ treatment at the end of the study. Improvements in disease activity, systemic symptoms and patient report outcomes were observed at the end of the study. 22.7%(90/396) patients experienced at least one treatment related adverse event, and 8 patients experienced at least one serious adverse event.Conclusion:This study demonstrates that TCZ treatment is effective in patients with RA when being treated for 6 months with an acceptable safety profile. The duration of TCZ treatment needs to be extended.

Objective: Abstract: Objective: To investigate the expression and significance of miR-138 and programmed cell death protein 1 (PD-1) in the patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Methods: A total of 30 patients with HBV-related HCC, 20 with HBV-related cirrhosis (LC) and 30 with chronic hepatitis B (CHB) were recruited from Jiaozuo People′s Hospital. The blood samples from all patients and the peritoneal effusion samples from HCC and LC patients were collected. The levels of miR-138 and soluble PD-1 (sPD-1) in blood and peritoneal effusion samples were detected by real-time PCR and ELISA, respectively. The expressions of PD-1 in T lymphocytes were measured with flow cytometry and western blot. The targeting effect of miR-138 on the 3′-non-coding region (3′-UTR) of PD-1 gene was verified by the dual-luciferase reporter gene system. Results: The relative expression levels of miR-138 in the peritoneal effusion and plasma of HBV-related HCC patients were significantly lower than those in LC and CHB patients (P＜0.05). The serum sPD-1 levels and the expression levels of PD-1 in CD3 + T lymphocytes of HBV-related HCC patients were significantly higher than those in LC and CHB patients (P＜0.05). The relative expression levels of miR-138 were negatively correlated with serum sPD-1 levels and the expression levels of PD-1 in CD3 + T lymphocytes (P＜0.05). The dual-luciferase reporter gene system and western blot results demonstrated that there was a targeting relationship between miR-138 and the 3′-UTR of PD-1 gene. After miR-138 was transfected, the expression level of PD-1 was significantly down-regulated. Conclusion miR-138 participates in the development and progression of HBV-related HCC probably by targeting PD-1.