Objective To explore the role of secreted phosphoprotein 1(SPP1)and myeloid differentiation primary response 88(MYD88)in morphine-induced cardiomyocyte apoptosis.Methods A morphine addiction model was established in Sprague-Dawley(SD)rats.Twelve SD rats were randomly assigned to the normal saline(NS)group or the morphine-dependent(DAM)group.Histopathological analysis was employed to observe and compare myocardial tissue morphology between the two groups.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was performed to assess the number of apoptotic cells in each group.The expression levels of SPP1 and MYD88 were evaluated using immunohistochemistry.Quantitative real-time poly merase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression of SPP1,MYD88,Bax,Bcl2,Caspase-3,and Caspase-9.Simultaneously,Western blot analysis was used to detected the expression of Cleaved Caspase-3 and Cleaved Caspase-9 proteins.In vitro,SPP1 expression was knocked down in primary neonatal rat cardiomyocytes(NRCMs),and cells were divided into three groups:control(CON),morphine treated(DA),and shSPP1#3+DA.Cell viability was assessed using the CCK-8 assay,and apoptosis rates were determined by flow cytometry.Results HE and TUNEL staining of myocardial tissues from morphine-addicted SD rats revealed that,compared with the NS group,myofibrils in the DAM group exhibited partial disruption and a significant increase in apoptotic cells(P<0.05).Western blot and RT-qPCR analyses demonstrated that,relative to the NS group,the mRNA and protein levels of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 were significantly upregulated in the DAM group(P<0.05),whereas Bcl2 expression was significantly downregulated at both mRNA and protein levels(P<0.05),and the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were also increased.with all differences being statistically significant.In NRCMs following morphine intervention,cell viability in the DA group was markedly reduced compared to the CON group(P<0.05),accompanied by a signifi-cant increase in apoptosis rate(P<0.05).Consistently,Western blot and RT-qPCR results showed elevated mRNA and protein expression of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 in the DA group(P<0.05),along with decreased Bcl2 expression(P<0.05).The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were elevated simultaneously.In contrast,the shSPP1#3+DA group exhibited opposing trends compared to the DA group,with statistically sig nificant differences(P<0.05).Conclusion SPP1 and MYD88 play critical roles in mediating morphine-induced cardiomyocyte apoptosis,and silencing SPP1 has been shown to significantly reduce the extent of cardiomyocyte apoptosis following morphine exposure.

Objective To explore the role of secreted phosphoprotein 1(SPP1)and myeloid differentiation primary response 88(MYD88)in morphine-induced cardiomyocyte apoptosis.Methods A morphine addiction model was established in Sprague-Dawley(SD)rats.Twelve SD rats were randomly assigned to the normal saline(NS)group or the morphine-dependent(DAM)group.Histopathological analysis was employed to observe and compare myocardial tissue morphology between the two groups.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was performed to assess the number of apoptotic cells in each group.The expression levels of SPP1 and MYD88 were evaluated using immunohistochemistry.Quantitative real-time poly merase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression of SPP1,MYD88,Bax,Bcl2,Caspase-3,and Caspase-9.Simultaneously,Western blot analysis was used to detected the expression of Cleaved Caspase-3 and Cleaved Caspase-9 proteins.In vitro,SPP1 expression was knocked down in primary neonatal rat cardiomyocytes(NRCMs),and cells were divided into three groups:control(CON),morphine treated(DA),and shSPP1#3+DA.Cell viability was assessed using the CCK-8 assay,and apoptosis rates were determined by flow cytometry.Results HE and TUNEL staining of myocardial tissues from morphine-addicted SD rats revealed that,compared with the NS group,myofibrils in the DAM group exhibited partial disruption and a significant increase in apoptotic cells(P<0.05).Western blot and RT-qPCR analyses demonstrated that,relative to the NS group,the mRNA and protein levels of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 were significantly upregulated in the DAM group(P<0.05),whereas Bcl2 expression was significantly downregulated at both mRNA and protein levels(P<0.05),and the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were also increased.with all differences being statistically significant.In NRCMs following morphine intervention,cell viability in the DA group was markedly reduced compared to the CON group(P<0.05),accompanied by a signifi-cant increase in apoptosis rate(P<0.05).Consistently,Western blot and RT-qPCR results showed elevated mRNA and protein expression of SPP1,MYD88,Bax,Caspase-3,and Caspase-9 in the DA group(P<0.05),along with decreased Bcl2 expression(P<0.05).The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were elevated simultaneously.In contrast,the shSPP1#3+DA group exhibited opposing trends compared to the DA group,with statistically sig nificant differences(P<0.05).Conclusion SPP1 and MYD88 play critical roles in mediating morphine-induced cardiomyocyte apoptosis,and silencing SPP1 has been shown to significantly reduce the extent of cardiomyocyte apoptosis following morphine exposure.

ObjectiveTo obtain the status of latent tuberculosis infection （LTBI） among tuberculosis （TB）-related health-care workers （HCWs） in Shanghai， and to explore the risk factors related to TB infection. MethodsA multi-center cross-sectional study was conducted by recruiting medical workers from multiple designated TB hospitals， centers for disease control and prevention， and community health service centers in Shanghai. Each subject was required to complete a questionnaire and to provide a blood sample for TB infection test. Univariate and multivariate analysis ware made in order to find risk factors relating to TB infection. ResultsA total of 165 medical workers were recruited， and the proportion of TB infection was 16.36% （95%CI： 11.49%‒22.76%）. Multivariate logistic analysis showed that clinical doctors and nurses （adjusted OR=9.756， 95%CI： 1.790‒53.188）， laboratory staffs （adjusted OR=78.975， 95%CI： 8.749‒712.918）， and nursing and cleaning workers （adjusted OR=89.920， 95%CI： 3.111‒2 598.930） had higher risk of TB infection. ConclusionThe overall LTBI prevalence among TB-related HCWs is low. However， working as doctors， nurses， laboratory staffs， nursing workers and cleaning workers are risk factors of TB infection. TB-related HCWs who work at hospitals are at risk of TB infection comparing to medical staffs who work outside hospitals.

Objective:To investigate the application value of oral stimulation combined with maternal milk sucking in the pain caused by PICC in children with congenital digestive tract malformations.Methods:Using convenience sampling method, 80 children with congenital digestive tract malformations who were admitted to Hunan Children's Hospital from July 2018 to December 2019 were selected and divided into the experimental group and the control group of 40 cases in each group according to the random number table method. The control group was given routine nursing mode, while the experimental group was given oral stimulation combined with maternal milk sucking intervention on the basis of routine nursing. The intervention effects were evaluated by the Neonatal Infant Pain Scale (NIPS), the onset time of the first cry, the duration of crying, the onset time of the first painful face, the duration of the painful face, and indicators such as heart rate, respiratory rate and SpO 2. Results:The NIPS scores of children in the experimental group were (5.60 ± 1.24) and (4.10 ± 0.31) in the puncture period and the recovery period, which were significantly lower than those in the control group (6.10 ± 0.84), (4.93 ± 0.35), the differences were statistically significant (t = 2.12, 3.00, both P<0.05). The heart rate and respiratory rate of the children in the experimental group were (151.10 ± 12.37), (147.00 ± 12.16) times/min and (47.48 ± 7.59), (45.58 ± 6.51) times/min in the puncture period and the recovery period, which were significantly lower than those in the control group (159.75 ± 13.81), (156.05 ± 12.99) times/min and (52.38 ± 8.12), (49.75 ± 7.59) times/min, the differences were statistically significant (t values were 2.64-3.22, all P<0.05). The SpO 2 in the experimental group were 0.980 5 ± 0.009 0, 0.982 5 ± 0.010 8 in the puncture period and the recovery period, which were significantly higher than those in the control group 0.970 8 ± 0.014 2, 0.971 8 ± 0.018 1, the differences were statistically significant (t = 3.66, 3.23, both P<0.05). The onset time of the first cry, the duration of crying, the onset time of the first painful face, the duration of the painful face in the experimental group were (41.55 ± 6.38), (22.05 ± 4.47), (35.08 ± 5.94), (24.65 ± 5.46) s, significantly shorter than those in the control group (46.58 ± 5.26), (29.50 ± 6.48), (44.68 ± 13.31), (30.65 ± 9.42) s, the differences were statistically significant (t values were 2.63-5.98, all P<0.05). Conclusions:Oral stimulation combined with maternal milk sucking can effectively relieve the pain caused by PICC in children with congenital digestive tract malformation, and reduce the physiological stress response of children.

Objective: To develop an instrument for measuring nursing outcomes classification in Chinese cerebral hemorrhage patient and test its reliability and validity. Methods: Based on Nursing Outcomes Classification, the primary item pool was formulated through literature review, semi-structured interviews and group meeting, the items were selected by expert consultation. Totally 186 elderly adults were selected to test the reliability and validity. Results: The final scale consisted of 5 dimensions and 34 items. The content validity index of the total scale was 0.912, the content validity index of each item was 0.833-1.000. Exploratory factor analysis extracted five common factors and the cumulative variance contribution rate was 63.189%; and the correlation coefficient between the total scale and each dimension were 0.760-0.874. The correlation coefficient of Stroke-Specific Quality of Life between total scale and each dimension were 0.789, 0.568-0.659. The Cronbach α was 0.960 for the total scale, the Cronbach α of dimensions were 0.858-0.932. The intraclass correlation coefficient of the scale and each dimension were 0.965, 0.885-0.935. Conclusions Cerebral Hemorrhage Patient Nursing Outcomes Classification Assessment Scale has good reliability and validity, which can be used to evaluate cerebral hemorrhage.

Objective: To investigate the application of music intervention in combination with maternal voice during PICC placement procedures in malformation alimentary tract infants. Methods: A total of 60 malformation alimentary tract infants were enrolled in the present study in Hunan Children Hospital from January to December 2018, infants were randomly assigned to the intervention group (n=30) and the control group (n=30). Patients in the control group received conventional nursing, the intervention group received music and maternal voice intervention method. The level changes of heart rate, respiratory rate, blood pressure, blood oxygen saturation and cortisol concentration were compared between two groups, meanwhile, the pain rating were assessed by Neonatal Infant Pain Scale (NIPS). Results: Before procedure, no significant difference was found between the control and intervention group in heart rate, respiratory rate, systolic pressure, SpO₂ and NIPS score (P>0.05). During and after the procedure, the mean heart rate [(144.43±15.93), (142.43±20.42) times/min], respiratory rate [(44.70±4.51) and (43.50± 4.12) times/min], systolic pressure [(87.40±8.53) and (86.10±4.60) mmHg] and NIPS score (5.34±0.71 and 5.19±0.87) of the intervention group was statistically lower compared to the control group [(160.97±39.35) and (159.17±31.87) times/min, (53.13±7.85) and (50.47±7.70) times/min, (97.60±9.82) and (94.27±7.23) mmHg, 6.17±0.54 and 6.15±0.66; during procedure, the mean SpO₂ was statistically significantly higher, compared to the control group [(96.15±9.82)% versus (88.34±9.53)%]. The difference was statistically significantly (t=2.134-6.682, P<0.05). Before placement, the concentration of cortisol between the two groups showed no significant difference (P>0.05), and (208.91±21.07)ng/ml 5min after placement and (197.70±16.34) ng/ml immediately after placement, the cortisol concentration was statistically significantly lower, compared to the control group [(246.71±35.15), (230.95±21.81) ng/ml] (t=5.050-6.682, P<0.05). Compared with the control group, the time needed for PICC placement decreased remarkably in the intervention group [(15.62±3.30) min versus (24.63±4.26) min)](t=9.160, P<0.01). Conclusions Music in combination with maternal voice can significantly reduce stress levels, pain responses and time need for PICC placement, music and maternal voice is a potential intervention strategy in malformation alimentary tract infants undergoing PICC.

Objective To investigate the correlation between iodine nutritional status and thyroid diseases in some areas of Ningxia. Methods The physical examination, questionnaire survey, and measurement of urine iodine, thyroid function, thyroid associated antibodies were performed, with thyroid ultrasonography in Jingyuan County, Xingqing District and Guyuan City, Ningxia Yinchuan City, totally including 2827 adults. Results The median urinary iodine of all subjects was 258. 7(76. 6-1506. 4) μg/L. The average level of urinary iodine in rural areas [203.3(64. 6-1154)μg/L] was lower than that in cities[340. 7(114. 8-1726. 1)μg/L, Z=19. 514, P>0. 01]. The average urinary iodine level was lower in people over 65 years old[234. 2(58. 9-2285. 4) μg/L, x2=6.449, P=0. 040]. Subjects in hypothyroidism group [232. 5(69. 2-1682. 3) μg/L] had the lowest average urinary iodine level, while hyperthyroidism group[331. 9(102. 4-1862) μg/L] turned to be the highest group( x2=15. 432, P>0. 01). Compared with the normal group, TPOAb and TGAb increased group had similar median levels of urinary iodine (P>0. 05). If compared with single nodule, multiple nodules group and the normal group's median levels of urinary iodine also had no significant difference(P>0. 05). Conclusion Iodine excess was found in the studied population in these areas of Ningxia.

Objective To assess the cognitive level of disease risk factors and clinical identification in high risk population of arteriosclerosis obliterans (ASO) of lower limbs. Methods By purpose sampling survey, a total of 330 patients in high risk of ASO from a ClassⅢ grade A hospital in Chongqing were investigated using a self-designed semi-structured questionnaire from May to June 2015 to assess the cognitive level of disease risk factors and clinical identification in high risk population of ASO. A database was set up in EpiData, and analyzed by SPSS 16.0 statistic software, including descriptive analysis, non-parametric test, Spearman correlation analysis. Results Among all respondents, 70.1% of them have never heard of lower limbs ASO. More than 80.0% of respondents did not know the risk factors, while more than 92.0% of them did not know its clinical manifestations, only 15.5% of them had received medical examination for diagnosing ASO. There were statistically significant differences in cognitive scores among respondents with different education background, residence and income (P< 0.01). Conclusions The cognitive level of disease risk factors and clinical identification in high risk population of ASO of lower limbs is obviously insufficient, especially those who are with lower education, lower income, and who live in small towns or villages. It is suggested that professional medical staff should strengthen the health education for people with ASO high-risk factors.

Objective To investigate the regulation mechanism of estrogen on the free calcium of smooth muscle cells at the endometrial-myometrial interface (EMI) in uteri with adenomyosis. Methods From September 2011 to November 2012, 59 uterine myometrial specimens were obtained from 59 cases underwent hysterectomy, including 28 adenomyosis patients as adenomyosis (ADS) group and 31 patients with cervical intraepithelial neoplasia Ⅲ as control group. EMI smooth muscle cells were cultured and loaded with calcium ion fluorescent probe fluo-4/AM. After treated with trisphosphate (IP3) receptor antagonist, blocker of sarcoplasmic reticulum calcium-adenosine triphosphate (ATP), depleted agent of the ryanodine receptor-operated Ca2+, inhibitor of L-type calcium channel, inhibitor of Na+-Ca2+exchanger, the labeled cells were stimulated with estrogen. The changes of intracellular Ca2+fluorescence intensity were detected by laser scanning microscopy. The changes of intracellular Ca2+concentration was indicated byΔF[Ca2+]i. Results (1) Under normal calcium conditions, after the stimulation of estrogen, intracellular Ca2+fluorescence intensity in ADS group and control group both increased than those without estrogen. TheΔF[Ca2+]i in ADS group was 384±26, and in the control groupΔF[Ca2+]i was 235±20. TheΔF[Ca2+]i in ADS group was higher than that in the control (P<0.01). Without calcium conditions, theΔF[Ca2+]i in ADS group was 207 ± 17, and in the control group ΔF[Ca2+]i was 221 ± 19. TheΔF[Ca2+]i in ADS group was almost the same with the increase in the control (P=0.731). The ΔF[Ca2+]i in ADS group was significantly decreased compared with the calcium condition (P<0.01). However, there was no significant difference in the control between with and without calcium conditions (P=0.060). (2) After treated with IP3 receptor antagonist, blocker of sarcoplasmic reticulum calcium-ATP, depleted agent of the ryanodine receptor-operated Ca2+, theΔF[Ca2+]i in both groups were significantly reduced (P<0.05), the increase in ADS group was significantly higher than that in the control (P<0.05). (3)After treated with inhibitor of L-type calcium channel, theΔF[Ca2+]i in ADS group was 211 ± 19, while in the control group ΔF[Ca2+]i was 203 ± 16, and there was no significantly increased intracellular Ca2 +in both groups (P>0.05). But, the ΔF[Ca2 +]i in ADS group was significantly reduced after treatment compared to before treatment, (211 ± 19 vs 384 ± 28; P=0.001). The increase in control group was almost the same with before (203±16 vs 234±22, P=0.141). (4) After treated with inhibitor of Na+-Ca2+exchanger, theΔF[Ca2+]i in ADS group was 357 ± 24 and in the controlΔF[Ca2+]i was 209±19. The increase in ADS group was significant higher than that in the control (P=0.000). Compared withΔF[Ca2+]i on the condition without treating with inhibitor of Na+-Ca2+exchanger,ΔF[Ca2+]i was 363±21 in ADS group andΔF[Ca2+]i was 237±20 in control group after treatment. When compared with before treatment, there was no significant difference in both groups (P>0.05). Conclusions The increase of intracellular Ca2+induced by estrogen at EMI smooth muscle cells in adenomyosis patients was mostly from the release of arcoplasmic reticulum, and also from the Ca2+influx controlled by L-type calcium channel. The increase of Ca2+inducing abnormal contraction of EMI muscle may have relationship with the development of adenomyosis.

BACKGROUNDEpstein-Barr virus (EBV) infection is one of the most important factors of nasopharyngeal carcinoma (NPC) endemic areas. Transcription of EBV-encoded non-polyadenylated RNAs (EBERs) are presented in most of NPC tumors. Exploring EBERs as a prognostic marker for NPC might further be informative about the biology and the progression of the disease. The aim of this study was to analyze the role of EBV latency in the clinical management of nasopharyngeal carcinoma (NPC), by detecting EBERs.

METHODSRNA in situ hybridization (ISH) for detecting EBERs was carried out on 908 NPC tumor tissues. Overall survival (OS) curves were analyzed with the Kaplan-Meier method and the Cox proportional-hazards regression models.

RESULTSThe median follow-up time was 70 months (1-120 months). Eight hundred and sixteen (89.9%) from a total of 908 consecutive NPC cases were found to be EBV-EBER positive. EBER-ISH staining revealed nuclear localization in NPC cells. In the Kaplan-Meier analysis for OS, high EBER expression levels in NPC patients were statistically significant positive prognostic factors for survival (log-rank, P = 0.022), especially in adults aged 17-40 years (P = 0.023) and in those with advanced stage disease (log-rank, P = 0.002). Cox proportional-hazards regression model analysis showed that the EBER expression level was an independent risk factor for OS (hazard ratio 0.724, P = 0.005).

CONCLUSIONSEBERs were frequently detected in NPC tumor tissues, and high-level EBER expression correlated with good prognosis in NPC patients, especially in adult patients and in those with advanced stage disease. EBER may serve as a potential prognostic predictor in NPC.

Adolescent ; Adult ; Aged ; Carcinoma ; Epstein-Barr Virus Infections ; virology ; Female ; Herpesvirus 4, Human ; genetics ; pathogenicity ; Humans ; In Situ Hybridization ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; mortality ; virology ; RNA, Viral ; genetics ; Young Adult