Objective To explore the effect of rotenone exposure on the metabolic homeostasis of nicotinamide adenine dinucleotide(NAD+)in dopaminergic neurons of the rat mid-brain striatum,and investigate the effect of exogenous NAD+intervention on the cellular damage response of dopaminergic neurons induced by rotenone.Methods Male SD rats(8 weeks old,200～250 g)were divided into a control group using a table of random numbers,a rotenone exposure group,an NAD+-intervention group,and an NAD+group.An intoxication model was established in the rotenone exposure group.NAD+(250 mg/kg)was administered simultaneously with rotenone exposure in the NAD+-intervention group.The NAD+group was only given NAD+,while the control group received no intervention.After modeling,open field test was performed to evaluate behavioral changes.After scarification,serum samples and mid-brain striatal tissues were collected.HE staining was used to observe the morphology of dopaminergic neurons in the striatum.The NAD+content in the tissues was detected with NAD+/NADH kit.Western blotting was employed to determine the contents of tyrosine hydroxylase(TH),nicotinamide phosphoribosyltransferase(NAMPT),nicotinamide mononucleotide adenylyltransferase(NMNAT),and solute carrier family 25 member A51(SLC25A51).ELISA was utilized to measure the content of dopamine in the striatal tissues.Immunohistochemical staining was applied to observe the distribution and contents of TH proteins in the striatal tissues of each group.Results Rotenone exposure significantly affected the vital signs and motor abilities of rats,induced disorderly-arranged,atrophy and deformed neurons in the striatal tissue,decreased the content of TH,rate-limiting enzyme for dopamine synthesis,by approximately 29%(P<0.01),the content of dopamine by about 42%,and that of NAD+by almost 50%(P<0.01),while increased the NADH/NAD+ratio(P<0.01).After exposure,the content of NAMPT,an enzyme related to NAD+synthesis,was decreased by 26%(P<0.05),the contents of NMNAT1-3 and SLC25A51,mitochondrial transporters of NAD+by approximately 21%,38%,43%,and 21%,respectively(P<0.01).Exogenous NAD+intervention improved the motor function of exposure rats and the morphology of dopaminergic neurons in the mid-brain striatal tissue,and restored the content of TH in the striatal tissue significantly by 12.8%(P<0.05),and the content of dopamine by 20.9%(P<0.05).Conclusion Rotenone disrupts the NAD+homeostasis in dopaminergic neurons by inhibiting the NAD+synthesis and transport pathways in the mid-brain striatal tissues,while exogenous NAD+intervention can effectively alleviate the dopaminergic neuron damage induced by rotenone exposure.

Breast cancer remains a leading cause of mortality in women worldwide.Triple-negative breast cancer(TNBC)is a particularly aggressive subtype characterized by rapid progression,poor prognosis,and lack of clear therapeutic targets.In the clinic,delineation of tumor heterogeneity and development of effective drugs continue to pose considerable challenges.Within the scope of our study,high hetero-geneity inherent to breast cancer was uncovered based on the landscape constructed from both tumor and healthy breast tissue samples.Notably,TNBC exhibited significant specificity regarding cell prolif-eration,differentiation,and disease progression.Significant associations between tumor grade,prog-nosis,and TNBC oncogenes were established via pseudotime trajectory analysis.Consequently,we further performed comprehensive characterization of the TNBC microenvironment.A crucial epithelial subcluster,E8,was identified as highly malignant and strongly associated with tumor cell proliferation in TNBC.Additionally,epithelial-mesenchymal transition(EMT)-associated fibroblast and M2 macrophage subclusters exerted an influence on E8 through cellular interactions,contributing to tumor growth.Characteristic genes in these three cluster cells could therefore serve as potential therapeutic targets for TNBC.The collective findings provided valuable insights that assisted in the screening of a series of therapeutic drugs,such as pelitinib.We further confirmed the anti-cancer effect of pelitinib in an orthotopic 4T1 tumor-bearing mouse model.Overall,our study sheds light on the unique characteristics of TNBC at single-cell resolution and the crucial cell types associated with tumor cell proliferation that may serve as potent tools in the development of effective anti-cancer drugs.

Objective:To establish the biology reference interval (RI) of peripheral blood procalcitonin (PCT) for children between 3 days and 6 years old in China.Methods:Totally 3 353 reference individuals with apparent health or no specific diseases were recruited in 18 hospitals throughout the country during October 2020 to May 2021. Reference individuals were divided into four groups: 3-28 days, 29 days - 1 year, 1-3 years and 4-6 years. Vein blood or capillary blood were collected by percutaneous puncture from every reference individual. The PCT level in serum and the capillary whole blood were assayed by Roche Cobas e601 and Norman NRM411-S7 immunoanalyzer. Outliers were deleted and 95th percentiles of every group were provided as RIs. Man-Whitney U test or Kruskal-Wallis test were used performed to assess the difference among different gender, age or method groups. Results:The difference of PCT distribution between male and female is not statistically significant, but the difference between serum and capillary whole blood is statistically significant. The differences between age groups are significant too. For Roche e601, serum PCT RI of 3-28 days group is <0.23 μg/L, 29 days - 6 years are <0.11 μg/L. For NRM411, Serum PCT RI of 3-28 days group is <0.21 μg/L, 29 days - 1 year: <0.09 μg/L, 1 - 6 years: <0.10 μg/L. For whole blood PCT, RI of 3-28 days group is <0.26 μg/L, 29 days - 6 years is <0.15 μg/L.Conclusions:Serum and capillary whole blood PCT have different RIs, however, capillary whole blood PCT testing is valuable in pediatric application. Children in 3-28 days show higher PCT levels than other age group. To establish the RIs and understand the differences among different groups are essential for the interpretation and clinical application of peripheral blood PCT testing results.

Objective To evaluate the effect of intelligentized patient-controlled analgesia ( PCA) management on the quality of postoperative analgesia in the patients. Methods A total of 6601 patients who underwent postoperative PCA from January 1, 2015 to December 31, 2017 searched from the intelli-gentized PCA system database were selected as intelligentized PCA management group ( I group) , and then were divided into 3 subgroups according to the year: 2015 subgroup ( n=2221 ) , 2016 subgroup ( n=2152) and 2017 subgroup (n=2228). A total of 1235 patients who underwent PCA which was mainly performed by a department of anesthesiology in the postoperative analgesia-related multi-center questionnaire from April 11, 2016 to April 22, 2016 in 12 grade A tertiary hospitals in Guangdong Province were select-ed as the traditional PCA management group (C group). The development of moderate and severe pain, nausea and vomiting, over-sedation at rest and during activity and patient′s satisfaction were recorded on 1st and 2nd days after operation. Results Compared with C group, the incidence of moderate and severe pain, nausea and vomiting and over-sedation at rest and during activity was significantly decreased, and the rate of patient′s satisfaction was increased at each time point after operation in I group ( P<0. 05 or 0. 01) . Com-pared with 2015 subgroup, the incidence of moderate and severe pain at rest and severe pain during activity was significantly decreased in 2016 and 2017 subgroups ( P<0. 05 or 0. 01) , and the incidence of nausea and vomiting was significantly increased in 2017 subgroup ( P<0. 05) . Compared with 2016 subgroup, the incidence of nausea and vomiting was significantly increased in 2017 subgroup (P<0. 05). Conclusion Intelligentized PCA management can improve the efficacy of PCA, mitigates the occurrence of adverse reac-tions and raise the quality of postoperative analgesia in the patients.

Objective To explore the clinical effect of continuous venous-venous hemofiltration (CVVH) combined with hemoperfusion (HP) in treatment of patients with hypertriglyceridemia pancreatitis (HTGP). Methods The clinical data of 33 patients with moderate and severe HTGP who were treated by CVVH combined with HP were retrospectively analyzed from March 2012 to March 2017 in Wuhan general hospital of the people's liberation army. The differences of vital signs and the serum levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), amylase (AMS), interleukin-6 (IL-6), blood calcium (Ca2+) and white blood cell count (WBC), haemoglobin (Hb), platelet count (PLT) before and 24 hours,72 hours and 1 week after therapy were compared, the changes of recovery time to target serum TG level, frequency of blood purification therapy, time for disease situation becoming stable, days staying in hospital and mortality were observed. Results The levels of LDL were not high in patients with HTGP, the levels of TG and TC were decreased significantly after using CVVH plus HP, and after treatment for 24 hours statistical differences appeared compared with those before treatment [TG (mmol/L):7.14±1.04 vs. 11.90±2.03, TC (mmol/L): 7.47±1.04 vs. 10.20±1.26], the decline persisting to 1 week after treatment;the drop rates of TG and TC were the largest after the first combined treatment, and the TG drop rate was more obvious than that of TC [(51.92±14.18)% vs. (30.09±10.01)%, P < 0.05], an average of (2.58±1.45) days and (2.38±0.98) times of combined blood purification could restore the TG to its safe level (TG < 5.65 mmol/L), the time of disease situation tending to be stable was (7.46±3.05) days and the time of staying in hospital was (20.00±2.12) days. Systemic inflammatory response syndrome (SIRS) related vital signs and inflammatory response indicators were also improved obviously after the combined therapy (all P < 0.05), after treatment for 72 hours, various vital signs and Ca2+reached to their normal reference ranges, after treatment for 24 hours IL-6 began to decline significantly compared with that before treatment (ng/L: 120.85±16.45 vs. 151.05±18.19), and AMS and WBC returned to their normal reference ranges after treatment for 1 week. Conclusion CVVH combined with HP can quickly and effectively eliminate TG in blood in patients with HTGP and in the mean time it may ameliorate and block the early progression of SIRS, resulting in good therapeutic effect on alleviating the disease development and improving its prognosis.

Objective To study the effects of hemodialysis (HD) combined with hemoperfusion (HP) on sleep quality in maintenance hemodialysis (MHD) patients. Methods Sixty MHD patients admitted to Department of Blood Purification of Wuhan General Hospital of PLA from January to December 2016, 30 cases were treated with HD, and the other 30 cases were treated by HD+HP, the course of treatment was 12 weeks in both groups. The changes of serum β2-microglobulin (β2-MG) and parathyroid hormone (iPTH) were observed before treatment and 12 weeks after treatment; the sleep quality of all patients in the two groups were evaluated by Pittsburgh Sleep Quality Index (PSQI) Scale, and the correlations between the sleep quality of MHD patients andβ2-MG level, iPTH level were analyzed by Pearson linear correlation analysis. Results All the 60 patients completed the treatment. The serum β2-MG, iPTH levels and PSQI score after treatment were decreased obviously in HD+HP group compared with those before treatment, and the degrees of decrease in HD+HP group were more significant than those in the HD group [β2-MG (mg/L): 12.34±2.12 vs. 20.27±3.15, iPTH (ng/L): 224.54±100.28 vs. 398.42±155.37, PSQI score:8.56±0.86 vs. 12.45±0.88, all P < 0.05]. Pearson linear correlation analysis showed that the PSQI score was significantly positively correlated with serum β2-MG, iPTH level (r respectively was 0.416 and 0.462, both P < 0.01). Conclusion HD+HP therapy can significantly improve the sleep quality of MHD patients, and the mechanism may be related to the elimination of serum iPTH and β2-MG from the body of MHD patients.

Objective]To investigate the change of spinal pro?inflammatory cytokines in a rat model of fentanyl induced acute hyperalgesia.[Methods]64 male SD rats were divided into 2 groups(n=32),fentanyl group and NS group. The rats were subcutaneously injected with fentanyl (60 μg/kg) or normal saline (1.2 mL/kg) 4 times with 15?minute intervals. Mechanical nociceptive thresholds and thermal nociceptive latency were measured via the tail pressure test(Tail flick thresholds,TFT) and paw withdrawal test(Paw withdrawal latency,PWL)on the day before,at 1,2,3,and 4 hour and on 1~5 day after injection. 4 rats were killed concomitantly and the lumber spinal cord were harvested to analysis the expression of NF-κB,PGE2,TNF-α,IL-1β,and IL-6.[Results]There were no significant changes of TFT,PWL and the expression of spinal inflammatory cytokines such as NF-κB, PGE2,IL-1β,and TNF-αcompared to baseline of rats treated with normal saline. The value of TFT ,PWL in fentanyl group raised to the highest(above the baseline)at the 1st hour after fentanyl injections and decreased thereafter,reached the lowest at the 1st day, raised increasinglyand up to baseline on the 3 day after injection. NF-κB,PGE2,IL-1β,and TNF-αincreased at the 4th hour,on 1 and 2 day and IL-6 increased at the 4 hour and onthe 1 day after fentanyl injections.[Conclusion]Subcutaneously injection of fentanyl induced significant mechanical and thermal hyperalgesia and increased spinal pro?inflammatory cytokines parallelly , indicated that fentanyl induced acute hyperalgesia is associated with spinal inflammatory reaction in rats.

Objective To observe the clinical curative effect of cinacalcet combined with activated vitamin D for treatment of patients with secondary hyperparathyroidism (SHPT) undergoing maintenance hemodialysis (MHD).Methods Eighty-six patients with SHPT undergoing MHD admitted to the Blood Purification Center of Wuhan General Hospital of Chinese People's Liberation Army from April 2014 to April 2016 were enrolled, and they were divided into an observation group and a control group by random number table, 43 cases in each group. The patients in control group were given cinacalcet whose initial dose was 25 mg/d and maximum dose should not exceed 75 mg/d, and the calcium acetate orally; on the basis of control group, the patients in observation group were additionally given activated vitamin D therapy, and both groups were treated for consecutive 12 weeks. After treatment, the clinical therapeutic effect, serum calcium, serum phosphorus, calcium phosphorus product, intact parathyroid hormone (iPTH) levels and the incidence of adverse reactions were compared between the two groups.Results The total effective rate in observation group was higher than that of the control group [90.70% (39/43) vs. 74.42% (32/43),P < 0.05]. After treatment, the difference of the serum calcium, calciumphosphorus product were higher than those before treatment in both groups [serum calcium (mmol/L): the control group was 2.24±0.25 vs. 1.99±0.26, observation group was 2.60±0.21 vs. 2.03±0.24; calcium phosphorus product (mmol2/L2): the control group was 4.05±0.34 vs. 3.79±0.35, observation group was 4.25±0.37 vs. 3.86±0.36, allP < 0.05], serum phosphorus, iPTH were lower than those before treatment in both groups [phosphorus (mmol/L): the control group was 1.69±0.14 vs. 2.09±0.12, observation group was 1.15±0.18 vs. 2.03±0.16; iPTH (ng/L): the control group was 297.36±59.73 vs. 499.54±69.32, observation group was 198.53±57.32 vs. 492.92±67.54, allP < 0.05], the degrees of changes in observation group were more significant than those in control group [serum calcium (mmol/L): 2.60±0.21 vs. 2.24±0.25, serum phosphorus (mmol/L): 1.15±0.18 vs. 1.69±0.14, calcium phosphorus product (mmol2/L2): 4.25±0.37 vs. 4.05±0.34, iPTH (ng/L): 198.53±57.32 vs. 297.36±59.73, allP < 0.05]; and the incidence of adverse reactions was significantly lower in observation group than that of the control group [4.65% (2/43) vs. 20.93% (9/43),P < 0.05].Conclusion Cinacalcet combined with activated vitaminD for treatment of SHPT patients undergoing maintenance hemodialysis shows obvious curative effect, reduces the whole segment of iPTH, and simultaneously has less adverse reactions.

Objective To investigate the clinical characteristics and antibiotic resistance of the bloodstream infections due to NDM-1 producing Klebsiella pneumoniae in children.Methods The nonduplicate carbapenem-resistant Klebsiella pneumoniae (CRKp) strains isolated from blood samples were collected in Beijing Children's Hospital from January 2011 to August 2014.Antimicrobial susceptibility was tested with broth microdilution method.PCR amplification and DNA sequencing were conducted targeting blaNDM-1 genes.Medical records were reviewed and analyzed.Results Of the 52 CRKp strains,blaNDM-1 gene was detected in 28 strains.All NDM-l-producing strains were multidrug-resistant.All the 28 isolates were resistant to penicillin,cephalosporins,piperacillin-tazobactam,and imipenem.More than 75.0％ of these NDM-1-producing strains were resistant to aztreonam,trimethoprim-sulfamethoxazole,gentamicin,and meropenem (92.9％,26/28).NDM-1-producing isolates had higher carbapenem MICs than non-NDM-1-producing isolates.Most (82.1％,23/28) of the NDM-1-producing isolates were isolated from hematology-oncology ward.The most common underlying disease was hematologic malignancy (78.6％,22/28).Febrile neutropenia was found in 20 (71.4％) patients.No difference was found between NDM-1-producing and non-NDM-1-producing CRKp infection in terms of repeated hospitalization (P=0.202),prior antibiotic use (P=0.615),underlying diseases (P=0.856),and deep venous catheter (P=0.099).After the susceptibility results were available,37 patients received carbapenembased combination regimen.The mortality did not show difference between NDM-1 producing CRKp infections and non-NDM-1-producing CRKp infections,7.1％ (2/28) vs.12.5％ (3/24),P=0.625.Conclusions The NDM-1 carbapenemase producing Klebsiella pneumoniae is emerging in this hospital.NDM-1-producing strains are resistant to multiple antimicrobial agents,associated with higher carbapenem MIC value.However,no difference was found in the clinical features between the bloodstream infections due to NDM-1-producing strain and those due to non-NDM-1-producing strains.

Objective To investigate the expression of pro-inflammatory cytokines in lumbar dorsal root ganglions (DRG) of rats model of high-dose fentanyl induced hyperalgesia. Methods 64 male SD rats were divided into 2 groups (n = 32), fentanyl group and normal saline (NS) group. The rats were injected with fentanyl (60 μg/kg) or NS 4 times in total subcutaneously with a 15-minute interval. Mechanical and thermal nociception were measured via the tail pressure test (tail flick thresholds, TFT) and paw withdrawal test (paw withdrawal latency, PWL) at 1 day before, at 1, 2, 3 and 4 hour and on 1 ~ 7 day after administration. 4 rats were sacrificed and the lumbar DRG were harvested to analyze the expression of PGE2 , IL-1β, IL-6 and TNF-αvia ELISA. Results There were no significant changes of TFT, PWL and the expression of pro-inflammatory cytokines in DRG compared to baseline of rats in NS group. The value of TFT , PWL in fentanyl group were above the baseline at the 1 ~ 4 hour and below the baseline at 1~3 day after fentanyl injections. PGE2 , IL-1β, TNF-α and IL-6 increased on 1,3,5,7 day after fentanyl injections significantly. Conclusions High-dose fentanyl induced significant hyperalgesia and up-regulation of pro-inflammatory cytokines in DRG. The expression pro-inflammatory cytokines peaked later and were more protracted than the change of behavior test and show no direct relationship between the two.