We previously identified a unique nucleus, the cerebrospinal fluid (CSF)-contacting nucleus. This study aims to understand its gene architecture and preliminarily suggest its functions. The results showed that there were about 19,666 genes in this nucleus, of which 913 were distinct from the dorsal raphe nucleus (non-CSF contacting). The top 40 highly-expressed genes are mainly related to energy metabolism, protein synthesis, transport, secretion, and hydrolysis. The main neurotransmitter is 5-HT. The receptors of 5-HT and GABA are abundant. The channels for Cl^-, Na⁺, K⁺, and Ca²⁺ are routinely expressed. The signaling molecules associated with the CaMK, JAK, and MAPK pathways were identified accurately. In particular, the channels of transient receptor potential associated with nociceptors and the solute carrier superfamily members associated with cell membrane transport were significantly expressed. The relationship between the main genes of the nucleus and life activities is preliminarily verified.
Rats ; Animals ; Rats, Sprague-Dawley ; Serotonin/metabolism* ; Signal Transduction ; Cerebrospinal Fluid/metabolism*

OBJECTIVE To establish a quantitative analysis method for the quality control components in Tenghuang jiangu capsules， and predict the possible action mechanism of the quality control components. METHODS Seven key quality control components in Tenghuang jiangu capsules were quantitatively analyzed by UHPLC-Q-Orbitrap HRMS. The “component-target” network was constructed based on network pharmacology， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and gene ontology （GO） function enrichment analysis were further conducted to find the key signaling pathways. RESULTS The average contents of succinic acid， hyperoside， gallic acid， kaempferol， naringin， naringenin and protocatechuic acid in 20 batches of Tenghuang jiangu capsules were 520.92， 67.67， 129.48， 4.74， 397.45， 5.66 and 376.62 μg/g， respectively. The results of network pharmacology showed that the 62 key target genes of the quality control components of the drug included AKT1， TNF， VEGFA， MMP9， PTGS2， etc. They were mainly enriched in cytokine receptor interaction， nuclear factor， tumor necrosis factor， interleukin 17， rheumatoid arthritis， Toll-like receptor and other signal pathways， involving inflammatory reaction， signal transduction， protein phosphorylation and other biological processes， kytoplasm， cell membrane and other cell components， as well as enzyme activity， energy activity and other molecular functions. CONCLUSIONS The established UHPLC- Q-Orbitrap HRMS method can be used for the quantitative analysis of the quality control components of Tenghuang jiangu capsule. Its quality control components may be mapped to inflammatory pathways related to bone diseases such as rheumatoid arthritis and Toll-like receptors through AKT1， TNF， VEGFA and other key targets， so as to play a therapeutic role.

Objective:To explore the protective effect of vagus nerve stimulation (VNS) on the prognosis of rats suffering from cardiac arrest/cardiopulmonary resuscitation (CA/CPR) under different treatment timings.Methods:The method of percutaneous epicardial electrical stimulation was used to establish CA model of rat. Fifty-three male SD rats were randomly (random number) divided into the sham group ( n=5), CPR group ( n=12), PRE group ( n=12), POST5 group ( n=12) and POST30 group ( n=12). The sham group did not experience CA/CPR. VNS treatment was started at 30 min before CA (PRE group, n=12), 5 min after recovery of spontaneous circulation (ROSC) (POST5 group, n=12), and 30 min after ROSC (POST30 group, n=12) in different VNS-treated group, respectively. The electrical stimulation was applied to the vagus nerve for 30 min with a unified parameter. The neurological deficit scores at 24, 48, and 72 h after ROSC were recorded, and the survival rate in each group was observed. TUNEL staining was used to detect the apoptosis of cortical area and the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in brain tissue was measured by immunofluorescence at 72 h after ROSC. Variables were compared with one-way analysis of variance, and survival for Kaplan-Meier curves were tested with the log-rank test. A P value less than 0.05 was considered statistically significant. Results:Compared with the CPR group (survival rate 33.33%), both pre-treatment (survival rate 75%) and post-treatment of VNS (POST5 group survival rate 75% and POST30 group survival rate 83.33%) significantly improved the 72 h survival rate after CPR ( P<0.05), mitigated neurological deficits after ROSC, reduced the positive rate of apoptosis neurons, and up-regulated the expression of α7nAChR in cerebral cortex. There was no significant difference among the VNS-treated groups (all P>0.05). Conclusions:Both pre-treatment and post-treatment of VNS can play a protective role in rats after CA/CPR, which may be related to the activation of α7nAChR-mediated anti-inflammatory and anti-apoptosis effects.

Objective To investigate the effects of precautionary rescue nursing intervention on the nursing quality score and efficacy of rescue for neonates with hypoxic ischemic encephalopathy. Methods One hundred children with neonatal hypoxic ischemic encephalopathy admitted to the First Hospital of Hebei Medical University from January 2017 to December 2018 were enrolled. Fifty children from January to December 2017 were assigned in the routine rescue nursing group; and 50 children from January to December 2018 were in the precautionary rescue nursing group. The routine rescue nursing group received routine rescue nursing; and the precautionary rescue nursing group was given pre-planned rescue nursing intervention. The rescue success rate, rescue time, Apgar score within rescue for 5 minutes, poor prognosis, etc. were recorded in both groups; the infant intelligence development scale was used to evaluate the situation of children's development of IQ in 6 months; a self-made questionnaire was used to investigate the rate of satisfaction in 100 children family guardians; the hospital's self-made Nursing Quality Assessment Form was applied to assess the quality of care of nursing staff. Results The rescue successful rate and the 5-minute Apgar score of the precautionary rescue nursing group were significantly higher than those of the routine rescue nursing group [rescue success rate: 98.0% (49/50) vs. 72.0% (36/50), Apgar score: 8.43±1.28 vs. 5.74±1.39], and the rescue time and poor prognosis rate were significantly lower than those in the routine rescue nursing group [rescue time (minutes):3.18±1.20 vs. 6.78±2.23, poor prognosis rate: 2.0% (1/50) vs. 14.0% (7/50), both P < 0.05]. The 6-month intelligence index and psychomotor index in the precautionary rescue nursing group were significantly higher than those in the routine rescue nursing group (intelligence index scores: 96.43±5.85 vs. 85.40±8.90, psychomotor index scores:97.38±4.03 vs. 78.43±10.22, both P < 0.05). The satisfaction rate in precautionary rescue nursing group was obviously higher than that in the routine rescue nursing group [92.0% (46/50) vs. 76.0% (38/50), P < 0.05]; the rescue technology, rescue efficiency, rescue effect, humanistic care score in precautionary rescue nursing group were significantly higher than those in the routine rescue nursing group (8.12±0.89 vs. 2.74±1.02, 8.23±1.24 vs. 3.01±0.83, 7.80±1.33 vs. 3.44±1.25, 8.03±1.32 vs. 3.29±1.38, respectively all P < 0.05). Conclusion The precautionary rescue nursing intervention used for neonates with hypoxic ischemic encephalopathy can significantly improve the rescue effect, degree of satisfaction to nursing and the quality of nursing care, so that it is worthy to be applied clinically.

Objective To establish the cardiac arrest (CA) model in rats by modified transcutaneous electrical stimulation on epicardium. Methods This study was performed in the Emergency Medicine laboratory in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. After 10 Sprague-Dawley male rats weighing 330-380 g were anesthetized, two acupuncture needles connected to the anode and cathode of a stimulator were transcutaneously inserted into the epicardium as electrodes. The puncture points were located quantitatively according to the anatomical structure of the rat chest. The electrical stimulation was maintained for 3 minutes to induce ventricular fibrillation(VF). Cardiopulmonary resuscitation (CPR) included chest compressions, intravenous adrenaline and defi brillation operated at 6 min after a period of nonintervention. Results CA was induced after the implement of the effective electrical stimulation in all ten rats in this experiment. The average current intensity to induce VF was (1.80 ± 0.59) mA, the average time to induce CA was (5.07 ± 2.37)s,the average time of the total electrical stimulation was(187.50 ± 12.75)s and the total time of CA was 6 min. At the end of the electrical stimulation, 9 rats presented VF and 1 rat showed pulseless electrical activity. The restoration of spontaneous circulation was achieved in all 10 rats. The average time of CPR was(190.90±68.60) s, the mean numbers of defi brillation were(1.20 ± 0.63) , and he average number of adrenaline application were (1.20 ± 0.42) times. Neither visible hemorrhage on epicardium nor gross pulmonary congestion was observed. Conclusions The modified transcutaneous electrical stimulation on epicardium to produce CA model in rats is an easily applicable and effective technique. This model may provide an alternative for experimental research of CPR.

Objective To evaluate the role of spinal histone acetylation in persistent postoperative pain in rats. Methods Pathogen?free healthy male Sprague?Dawley rats, weighing 200-250 g, aged 2 months, in which intrathecal catheters were implanted at the lumbar level according to an improved method, were used in the study. Eighty?four rats, in which intrathecal catheters were successfully implanted, were divided intoⅠ-Ⅵgroups(n=14 each)using a random number table. Artificial cerebrospinal fluid 20 μl was intrathecally administered at 1, 2, 3 and 4 days before operation and 1 day after operation inⅠandⅣgroups. At 1, 2, 3 and 4 days before operation and 1 day after operation, dimethyl sulfoxide 10 μl and SAHA(50 μg∕10μl)were intrathecally injected inⅡandⅤgroups and inⅢandⅥgroups, respective?ly, followed by artificial cerebrospinal fluid(10 μl)flush after each injection. Rats underwent sham oper?ation inⅠ?Ⅲ groups. Persistent postoperative pain was evoked by skin∕muscle incision and traction in Ⅳ?Ⅵ groups. The mechanical paw withdrawal threshold(MWT)was measured at 1 day before operation(T0)and 1, 3, 7, 14 and 21 days after operation(T1?5). Four rats were sacrificed in each group after measurement of MWT at T4, and the lumbar segments(L4?6)of the spinal cord were removed for determi?nation of the expression of acetylated histone H3(Ac?H3)and Ac?H4 by Western blot. Results There was no significant difference in each index amongⅠ?Ⅲ groups(P>0.05). Compared with group Ⅰ, the MWT was significantly decreased at T2?5, and the expression of Ac?H3 and Ac?H4 was down?regulated at T4 in group Ⅳ(P<0.05). Compared with group Ⅳ, the MWT was significantly increased at T2?5, and the expression of Ac?H3 and Ac?H4 was up?regulated at T4in group Ⅵ(P<0.05). Conclusion Histone acetylation is involved in the development and maintenance of persistent postoperative pain in rats.

Objective To study the application effect of systematic emergency nursing process in treatment of neonatal hypoxic ischemic encephalopathy (HIE). Methods One hundred and four neonates with HIE who were treated in the First Hospital of Hebei Medical University from January 2016 to January 2018 were selected as the subjects of this study, and they were divided into a control group and a study group according to different nursing care process, with 52 cases in each group. The infants in the control group were resuscitated according to conventional traditional first aid measures, while the infants in the study group received systematic emergency nursing care process on the basis of routine first aid (including the preparation of early rescue drugs and equipment; warmth kept in the whole course, the room temperature controlled in 30 - 32 ℃; rapid assessment of maternal delivery methods and procedures, delivery of the newborn and assessment of babies general conditions; performance of preliminary resuscitation for neonate, tracheal intubation for mechanical ventilation, and external chest compression if necessary; close monitor of the vital signs of newborn; symptomatic treatment; reassessment of the baby's disease condition). The successful rescue rate, total rescue time, hospitalization time, neonatal Apgar score and quality of nursing care before and after resuscitation were compared between the two groups. Results The successful rescue rate of the study group was higher than that of the control group [96.2% (50/52) vs. 82.7% (43/52), P < 0.05], and the total rescue time (minutes: 8.9±2.1 vs. 12.4±3.2) and hospitalization time (days: 8.1±2.2 vs. 11.4±2.9) in study group were shorter than those in control group, all the differences being statistically significant (all P < 0.05). After resuscitation for 5 minutes, the Apgar scores of the newborns in the two groups were higher than those before the resuscitation (study group: 8.3±1.1 vs. 2.2±0.7, control group: 6.7±2.0 vs. 2.1±0.9), and after resuscitation for 5 minutes, the Apgar score of the study group was higher than that of the control group (8.3±1.1 vs. 6.7±2.0), the difference being statistical significant (P < 0.05). The scores of rescue technique (5.0±2.9 vs. 2.7±0.6), humanistic care (5.6±2.1 vs. 2.8±0.9), rescue efficiency (5.3±1.2 vs. 3.0±0.6), and rescue effect (5.7±1.3 vs. 2.9±0.8) in the study group were all higher than those in the control group (all P < 0.05). Conclusion The systematic emergency nursing care process can effectively elevate the successful rescue rate of neonates with HIE, shorten the total rescue time and hospitalization time, enhance the rescue effect, and improve the quality of nursing care, thus it is worthy to be widely applied clinically.

Objective To investigate the changes in acetylation of histone in the spinal dorsal horn in a rat model of persistent postoperative pain.Methods Ninety-six malc Sprague-Dawley rats,weighing 200-250 g,aged 6-8 weeks,were randomly divided into 2 groups (n=48 each) using a random number table:sham operation group (group S) and persistent postoperative pain group (group PPP).The rat model of persistent postoperative pain evoked by skin/muscle incision and retraction was established according to the method described by Flatters.After the rats were anesthetized with intraperitoneal chloral hydrate,the skin and superficial muscle of the medial thigh were incised and retractors inserted.This tissue was retracted for 1 h.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before operation and 1,3,7,14,and 21 days after operation.Four animals were sacrificed in each group after measurement of MWT at each time point for detection of acetylated histone H3 (Ac-H3) and acetylated histone H4 (Ac-H4) expression (by Western blot analysis) and the number of Ac-H3 and Ac-H4 positive cells in the spinal cord horn (by immunofluorescence histochemistry).Results Compared with group S,the MWT was significantly decreased at 3,7,14 and 21 days after operation,the expression of Ac-H3 and Ac-H4b was significantly down-regulated at 3,7 and 14 days after operation,and the number of Ac-H3 and Ac-H4 positive cells was significantly decreased at 7,14 and 21 days after operation in group PPP (P＜0.05 or 0.01).The MWT,expression of Ac-H3 and Ac-H4b,and the number of Ac-H3 and Ac-H4 positive cells were significantly higher at 21 days after operation than at 14 days after operation in group PPP (P＜0.05).Conclusion Acetylation of histone in the spinal dorsal horn is decreased after operation,which may be involved in the development and maintenance of persistent postoperative pain in rats.

Objective To evaluate the effects of hydrogen-rich saline on chronic inflammatory pain in rats.Methods Thirty-two male Sprague-Dawley rats,aged 8-10 weeks,weighing 200-250 g,were randomly divided into 4 groups (n =8 each) using a random number table:control group (group C);complete Freund's adjuvant (CFA) group;hydrogen-rich saline group (group H2);CFA + hydrogenrich saline group (CFA+H2 group).Chronic inflammatory pain was induced by injecting CFA 100 μl into the plantar surface of the left hindpaw in CFA and CFA + H2 groups.In H2 and CFA + H2 groups,0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected intraperitoneally once a day for 7 consecutive days starting from 1 day after injection of CFA,while the equal volume of normal salinc was given instead of hydrogen-rich saline in C and CFA groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CFA injection and 1,3 and 7 days after CFA injection.The rats were sacrificed after the last measurement of pain threshold on day 7 after CFA injection.The left lumbar segments (L4 5) of the spinal cord were removed for determination of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression by Western blot.Results Compared with C group,no significant change was found in the MWT,TWL and expression of Nrf2 and HO-1 in H2 group,and the MWT was significantly decreased,the TWL was shortened,and the expression of Nrf2 and HO-1 was up-regulated in CFA and CFA+H2 groups.Compared with CFA group,the MWT was significantly increased,the TWL was prolonged,and the expression of Nrf2 and HO-1 was up-regulated in CFA+H2 group.Conclusion Hydrogen-rich saline can alleviate chronic inflammatory pain in rats,and activation of Nrf2/ARE signaling pathway in the spinal cord is involved in the mechanism.

Background and purpose:Pterostilbene is a natural antioxidant, whose role in retinoblastoma remains unclear. The aim of this study is to probe the effects of pterostilbene on the proliferation, apoptosis and autophagy in retinoblastoma WERI-Rb-1 cell lines.Methods:Cell counting kit-8 (CCK-8) assays were used to analyze the effects of pterostilbene on the proliferation of WERI-Rb-1 cells. Apoptosis rate was determined by Annexin V/PI. Autophagic vacuoles were observed by acridine orange staining. LC3 and P62 protein expressions were determined using Western blot.Results:Pterostilbene significantly inhibited the proliferation of WERI-Rb-1 cells (P<0.01). The cell viability were (93.02±0.47)%, (55.10±2.04)% and (30.33±1.45)% after WERI-Rb-1 cells were treated with 25, 50 and 100 μmol/L pterostilbene for 24 h, and the cell viability were (88.38±3.70)%, (53.37±1.17)%, (29.60±1.05)% after WERI-Rb-1 cells were treated with 50 μmol/L pterostilbene for 12, 24 and 48 h. Pterostilbene induced cell apoptosis (P<0.01), the apoptosis rates of control group, 24 h treated group and 48 h treated group were (4.08±0.79)%, (13.44±2.12)% and (23.49±2.01)%. Pterostilbene induced autophagy of WERI-Rb-1 cells, increased LC3 expression, downregulated P62 expression and increased the number of autophagic vacuoles in WERI-Rb-1 cells (P<0.01). 3-MA and Beclin1 were able to rescue pterostilbene-induced cell death (P<0.01). After 3-MA was used to blunt autophagosome formation, the apoptosis rate markedly decreased in 3-MA+pterostilbene-treated cells compared with cells treated with pterostilbene alone [(12.97±2.09)%vs (8.35±1.11)%], and after siRNA was used to knockdown Beclin1, the apoptosis rate had the same change [(13.80±2.19)%vs (9.62±0.52)%].Conclusion:Pterostilbene can inhibit the proliferation of WERI-Rb-1 cells and induce cell apoptosis via autophagy activation.