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:Myelodysplastic syndrome has worse hazards of acute myeloid leukemia transformation,and some studies have revealed that immune infiltration plays a vital part in the two.Nevertheless,more studies are required to confirm the relationship between immune infiltration and related differentially expressed gene regulation. OBJECTIVE:To screen the differentially expressed genes with prognostic significance between myelodysplastic syndrome and acute myeloid leukemia by bioinformatics analysis and explore the possible roles and mechanisms among these differentially expressed genes and immune infiltration mechanisms in the occurrence and progression of diseases. METHODS:The differentially expressed genes were screened for bioinformatics analysis using the GEO datasets,and analyzed by DO,GO,KEGG and GSEA.The TCGA prognostic database was used to plot the K-M curves of differentially expressed genes and receiver operating characteristic curve analysis was applied to evaluate the clinical diagnostic performance.Finally,CIBERSORT analysis was used to intuitively demonstrate the correlation between critical prognostic genes and the distribution of immuno-infiltrated cells.RT-qPCR was employed to detect peripheral blood samples from healthy controls,myelodysplastic syndrome and acute myeloid leukemia patients so as to verify the crucial genes preliminarily. RESULTS AND CONCLUSION:(1)A total of 150 differentially expressed genes were obtained between myelodysplastic syndrome and acute myeloid leukemia,among which 16 genes were up-regulated and 134 were down-regulated.(2)The results of DO,GO,KEGG and GSEA analysis suggested that differentially expressed genes might promote the development of myelodysplastic syndrome to acute myeloid leukemia by regulating the immune response.CIBERSORT revealed the differences in immune infiltration between myelodysplastic syndrome and acute myeloid leukemia.The distribution of CD4+ T cells,monocytes,neutrophils and M1 macrophages decreased in acute myeloid leukemia patients.In contrast,the distribution of inflammatory suppressor cells M2 macrophages increased,suggesting that it may be related to the immunosuppression of acute myeloid leukemia.(3)K-M curve and receiver operating characteristic curve analysis of 150 differentially expressed genes screened out four genes relevant to immunity and prognosis with good diagnostic performance:MANSC1,FLT3,BMX and CXCR2.(4)The results of RT-qPCR exhibited that MANSC1,BMX and CXCR2 were low expressed,while FLT3 was highly expressed in acute myeloid leukemia patients.These findings verify that the differential expression of MANSC1,FLT3,BMX and CXCR2 in patients with myelodysplastic syndrome and acute myeloid leukemia is not only significantly correlated with the prognosis of patients but may also affect the occurrence and development of myelodysplastic syndrome and acute myeloid leukemia by regulating the immune infiltration of patients.They can be used as potential biomarkers and therapeutic targets of the transformation from myelodysplastic syndrome to acute myeloid leukemia,providing a new direction for clinical diagnosis and treatment of the transformation of myelodysplastic syndrome.

Objective:To investigate the expression and significance of endoplasmic reticulum stress apoptosis pathway related proteins in renal cortex of rats with chronic fluorosis.Methods:Twenty four healthy SD rats were divided into 4 groups (6 rats/group, half male and half female) according to their body mass (100 - 120 g) by random number table method, rats in control group drank tap water (fluoride content < 0.5 mg/L), and in low, medium and high fluoride groups drank tap water with fluoride content (sodium fluoride) of 10, 50 and 100 mg/L, respectively. After 180 days of feeding, dental fluorosis was examined, 24-hour urine sample was collected and the content of fluoride in urine was detected by fluoride ion selective electrode method. Renal tissue was taken after anesthesia, and the pathological changes of renal cortex were observed by hematoxylin-eosin (HE) staining. The expressions of endoplasmic reticulum stress apoptosis pathway related proteins [inositol-requiring enzyme 1α (IRE1α), apoptosis signal-regulating kinase 1 (ASK1), C-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK)] were determined by immunohistochemical staining in rat renal cortex.Results:No dental fluorosis was found in control group. The incidence of dental fluorosis in low, medium and high fluoride groups were 2/6, 5/6 and 6/6, respectively. Compared with control group [(5.707 ± 1.190) mg/L], the urinary fluoride in low, medium and high fluoride groups [(17.028 ± 3.006), (34.378 ± 12.045), (94.759 ± 31.773) mg/L] was significantly higher ( P < 0.05), and the urinary fluoride in high fluoride group was higher than that in low and medium fluoride groups ( P < 0.05). HE staining showed that, compared with control group, the cell volume of renal tubules and glomeruli in medium and high fluoride groups increased, the cells arranged closely, and the eosinophilia of the cytoplasm increased. The immunohistochemical staining results showed that there was no significant difference in the expression of JNK protein in rat renal cortex between control group and low, medium and high fluoride groups ( F = 0.07, P > 0.05). The expressions of IRE1α, ASK1 and P-JNK proteins in rat renal cortex in high fluoride group were higher than those of control, low and medium fluoride groups ( P < 0.05), and the expressions of IRE1α and ASK1 proteins in medium fluoride group were significantly higher than those in control and low fluoride groups ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to renal cortex injury in rats, and the mechanism of injury may be related to the activation of IRE1α-ASK1-JNK endoplasmic reticulum stress apoptosis pathway.

Animals ; Mice ; Motor Neurons/physiology* ; Spinal Cord

Objective:To observe the change of growth and development, learning and memory, and oxidative stress in serum of offspring rats with fluorosis, and to explore the mechanism of fluoride on neurobehavioral development of offspring rats.Methods:Seventy-two SD rats (female and male ratio 3 ∶ 1) were fed adaptively for one week. According to their body mass [(80 ± 20) g], they were divided into control group (drinking tap water, containing less than 0.5 mg/L fluoride), low fluorine group (drinking water containing 10.0 mg/L of fluoride), and high fluorine group (drinking water containing 100.0 mg/L of fluoride) with random number table. After six months of feeding, they mated freely and gave birth in each group (24 rats with 18 females and 6 males). The rats in each group continued to be exposed to fluoride after giving birth, and the offspring rats were exposed to fluoride through breast milk feeding until the 28th day after birth. Body and brain weight, growth and development indicators (auricle separation, eyes opening, teeth eruption and hair growth) and neurobehavioral development indicators (cliff avoidance, auditory startle, surface righting and vibrissa positioning) were recorded. On the 28th day after birth, the learning and memory abilities (escape latency) of offspring rats were tested by Morris water maze; blood samples were taken from eyeballs to detect the content of nitric oxide (NO), the activity of nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS).Results:On the 21st day and 28th day after birth, the differences of body weight among control group, low fluorine group and high fluorine group [21st day: (54.70 ± 3.02), (52.30 ± 2.58), (51.30 ± 2.71) g, 28th day: (91.70 ± 5.03), (90.40 ± 4.76), (86.00 ± 4.55) g] were statistically significant ( F = 3.96, 3.70, P < 0.05); on the 21st day, the body weight of high fluorine group was lower than that of control group ( P < 0.05); on the 28th day, the body weight of the high fluorine group was lower than those of control group and low fluorine group ( P < 0.05). On the 28th day, the difference of brain weight of control group, low fluorine group and high fluorine group was statistically significant ( F = 6.19, P < 0.05); and the low fluorine group and high fluorine group were lower than that of control group ( P < 0.05). Among the growth development indicators, the difference of time of completing eyes opening in control group, low fluorine group and high fluorine group was statistically significant ( F = 3.64, P < 0.05); and the high fluorine group was higher than that of control group ( P < 0.05). In neurobehavioral development indicators, the differences of time of completing cliff avoidance, surface righting between the control group, low fluorine group and high fluorine group were statistically significant ( F = 8.29, 7.69, P < 0.05); and the time of completing cliff avoidance in high fluorine group was higher than those of control group and low fluorine group ( P < 0.05), the time of completing surface righting was higher than that of control group ( P < 0.05). In Morris water maze, on the 4th day, the escape latencies of low fluorine group and high fluorine group were higher than that of control group ( P < 0.05). The results of oxidative stress in serum showed that there were statistically significant differences in serum NO content, NOS and iNOS activitives between the control group, low fluorine group and high fluorine group ( F = 4.86, 66.48, 70.95, P < 0.05); and the NO content of the high fluoirne group was higher than that of the control group ( P < 0.05), the activities of NOS and iNOS of the high fluoirne group were higher than those of control group and the low fluorine group ( P < 0.05). Conclusion:Excessive fluoride can increase the level of oxidative stress in serum, which may be closely related to the neurobehavioral retardation and the decline of learning and memory ability of offspring rats.

Objective:To observe the changes of phosphorylated N-methyl-D-aspartate receptor (P-NMDAR) subunit and calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) protein expression in the brain tissue of rats with chronic fluorosis, and to explore the molecular pathogenesis of chronic fluorosis nerve injury.Methods:Eighteen one-month-old SD rats (half male and half female), weighing (100 ± 20) g, were randomly divided into three groups after adaptive feeding for 1 week: control group (drinking tap water, fluoride content < 0.5 mg/L), low fluoride group (drinking water fluoride content was 10.0 mg/L) and high fluoride group (drinking water fluoride content was 100.0 mg/L), with 6 rats in each group (half male and half female). Brain tissue was harvested after 180 days of feeding. HE staining was used to observe the morphological changes of rat brain tissue, Nissl staining was used to observe the changes of Nissl bodies in nerve cells, and immunohistochemical staining was used to observe the expressions of P-NMDAR subunits (P-NMDAR1, P-NMDAR2A) and CaMK Ⅱ protein in rat brain tissue.Results:Under light microscope, HE staining showed disordered arrangement of hippocampal neurons, nuclear hyperstaining and basophilic enhancement in the high fluoride group. The results of Nissl staining showed that the average optical density of Nissl bodies in nerve cells in the hippocampus of rats in the control group, low fluoride group, and high fluoride group were 0.024 4 ± 0.009 7, 0.024 0 ± 0.003 1, and 0.023 9 ± 0.013 8, respectively. There was no statistically significant difference between the groups ( F = 0.010, P > 0.05). Compared with the control group (0.011 8 ± 0.006 5, 0.065 6 ± 0.011 1, 0.143 8 ± 0.029 9) and low fluoride group (0.017 2 ± 0.006 8, 0.062 6 ± 0.017 8, 0.135 6 ± 0.029 6), the protein expressions of P-NMDAR1, P-NMDAR2A and CaMK Ⅱ in high fluoride group were significantly increased (0.026 3 ± 0.005 7, 0.086 3 ± 0.009 0, 0.210 9 ± 0.048 7, P < 0.05); and the protein expression of P-NMDAR1 in low fluoride group was higher than that in the control group ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to nerve cell injury in rats, and the mechanism of injury maybe related to the excitotoxicity induced by calcium ion (Ca 2+) overload caused by overactivation of NMDAR subunits.

Objective To retrieve and analyze the evidences on insulin injection position selection and rotation in patients with diabetes, and to summary the best evidence. MethodsWe took "diabetes mellitus""insulin" "injection" "position" "rotation" as subject terms to retrieve the evidences on insulin injection position selection and rotation in the evidence-based database such as the British Medical Journal (BMJ) best practice, Uptodata, Cochrane Library (2014), Joanna Briggs Institute Library, Registered Nurses' Association of Ontario (RNAO), Scottish Intercollegiate Guidelines Network (SIGN) and National Guideline Clearinghouse (NGC) as well as the comprehensive database such as the Embase, Elsevier science Direct, PubMed, Web of Science, OVID, China Biological Medicine (CBM) and WanFang data. ResultsA total of 13 evidences were included, 2 evidence summaries, 7 guidelines and 4 consensus. Four kinds of evidence were summarized including training for insulin standardized injection and management, evaluation, injection position and rotation. ConclusionsMedical staff should standardize the clinical practice behavior in insulin injection position selection and rotation based on relevant levels of evidence-based medicine to guarantee the safety of patient and improve quality of nursing.

Objective To investigate the serum 25-hydroxy vitamin D concentrations and related factors in early pregnancy. Methods Plasma was collected in the first trimester from 23 396 pregnant women to inves-tigate the vitamin D level, and its distribution and differences in different age, body maxx index ( BMI) and seasons between primipara and multipara. Preterm birth was used as an indicator of clinical outcomes. Vitamin D concentrations were measured using chemiluminescence microparticle immunoassay. Results 25-hydroxy vi-tamin D level was 42. 0 (17. 6-76. 6) nmol/L in totally 23 396 pregnant women with early pregnancy, and 5% and 95% percentile sites of vitamin D level were 20. 2 nmol/L and 70. 7 nmol/L respectively. There were 18 170 (77. 7%) primiparas and 5 226 (22. 3%) multiparas, with the mean age of 30. 0 (24. 0-38. 0) years and BMI of 20. 7 (16. 5-27. 6) kg/m2. The number of cases detected in spring (March, April and May), summer (June, July and August), autumn (September, October and November months) and winter (Decem-ber, January and February) were 5 878, 5 554, 5 974, and 5 990, respectively, and the vitamin D levels were 40. 0 (29. 3, 52. 7) nmol/L, 46. 2 (35. 6, 57. 2) nmol/L, 43. 8 (33. 1, 54. 8) nmol/L and 37. 2 (26. 9, 49. 9) nmol/L respectively, with the difference in vitamin D levels statistically significant among the four seasons (P<0. 001). According to BMI, all pregnant women were divided into four groups as BMI<18. 5 kg/m2, 18. 5 kg/m2≤BMI≤23. 9 kg/m2, 24 kg/m2≤BMI≤27. 9 kg/m2, BMI≥28 kg/m2, and the levels of 25-hydroxy vitamin D were 43. 5 (30. 9, 56. 9) nmol/L, 42. 1 (30. 8, 53. 8) nmol/L, 39. 9 (30. 7, 50. 4) nmol/L and 39. 7 (30. 7, 49. 4) nmol/L respectively with the difference statistically significant among the four groups. The levels of vitamin D detected in pregnant women with age<25 years, 25~29 years, 30~34 years,≥35 years were 39. 1 ( 28. 4, 52. 3) nmol/L, 41. 3 ( 30. 1, 52. 9) nmol/L, 42. 4 ( 31. 2, 54. 1) nmol/L and 43. 8 (31. 9, 55. 7) nmol/L respectively and the difference was statistically significant (P<0. 001). The levels of 25-hydroxy vitamin D in primary and multiparas were 41. 6 (30. 2, 52. 9) nmol/L and 43. 5 (32. 5, 56. 8) nmol/L with the difference statistically significant (P<0. 001). Among women of different gestational age during childbirth, the differences in serum 25-hydroxy vitamin D in early pregnancy were not statistically significant (P=0. 121). The severe deficiency of vitamin D in early pregnancy was defined as serum level of 25-hydroxy vita-min D less than 5th level. There were statistical differences in the probability of severe vitamin D deficiency of dif-ferent ages, seasons and BMI between primiparas and multiparas. Conclusions Pregnant women of lower weight, lower age and primiparity have higher incidence of the severe vitamin D deficiency in early trimester of pregnancy. There is significant difference in 25-hydroxy vitamin D level among the different seasons (winter

Objective To explore the effects of continuous nursing on postoperative urinary incontinence in patients with prostate cancer, so as to provide clinical evidence to improve the quality of postoperative and extended nursing care.Methods A total of 120 patients with prostate cancer underwent radical prostatectomy in the First Affiliated Hospital of Wenzhou Medical University from January 2016 to January 2017, who met the inclusion criteria were recruited in the research. The patients were randomly divided into experimental group and control group by sortition randomization method, with 60 cases in each. The patients in control group were given routine discharge health education, and the experimental group received continuous nursing on the basis of routine discharge nursing. Three months after discharge, the incidence, duration, frequency and quantity of urinary incontinence in the two groups were recorded by the International Consultation on Incontinence Questionnaire Short Form (ICI-Q-SF). A self-designed prostate cancer postoperative-related knowledge questionnaire, which evaluated the patients understanding prostate cancer care, was completed by the patients of the two groups. A self-designed satisfaction questionnaire was used to evaluate and compare the satisfaction of the two groups of patients to the quality of nursing.Results The incidence rate and the duration of postoperative urinary continence in the experimental group [16.7％,(4.1±2.8) d] were significantly lower than those in the control group (χ2=9.076,t=-2.630;P<0.05). After 3 months of discharge, the score of related knowledge and the satisfaction of nursing work in the experimental group were both higher than those of the control group, and the differences were statistically significant (P<0.05).Conclusions Continuous nursing care for discharged patients after prostatectomy can enhance patients' knowledge of disease, effectively reduce the incidence of postoperative incontinence, reduce urinary incontinence duration, and improve patients' satisfaction with nursing quality.

Objective To explore the real psychological experience in elderly patients with prostatic cancer after castration and to offer reference for formulating targeted interventions.Methods Totally 15 elderly prostatic cancer patients who received castration and total androgen blockade in the First Affiliated Hospital of Wenzhou Medical University from October 2014 to June 2016 were selected by convenient sampling and interviewed intensively.The data were analyzed by Colaizzi's seven-step method for qualitative studies.Results Totally 3 topics were summarized:the sense of uncertainty in elderly prostatic cancer patients after castration;changes in social adaptation;self-image disorder.Conclusions Various psychological crises,especially depression,are experienced in elderly prostatic cancer patients after castration.Therefore,continuous and extensive support is required for elderly prostatic cancer patients after castration.