AIM:To investigate the role of reactive oxygen species (ROS) in the regulation of intracellular Ca2+induced by angiotensin II ( Ang II) in the primarily cultured medullary neurons .METHODS:Primarily cultured me-dullary neurons were prepared from 14-day-old embryos of Sprague-Dawley rats in the study .The identification of medullary neurons was assessed by double-labeling immunofluorescence .To explore the role of ROS , mainly the superoxide ( O2 ·-) , the O2 ·-generation was measured using the fluorogenic probe dihydroethidium ( DHE) .To determine intracellular free cal-cium concentration ( [ Ca2+] i ) , the neurons were loaded with the Ca 2+-specific dye Fura-2/AM.The cell viability after adding Ang II was also examined using CCK-8 assay.RESULTS:Most of the cultured cells were medullary neurons , more than 80%of which were glutamate positive neurons .Ang II (5 μmol/L) increased the level of ROS within 10 min in the medullary neurons .Ang II at 5μmol/L induced a significant [ Ca2+] i increase in the medullary neurons , and the effect of Ang II occurred rapidly and reached a peak within 20 min after administration.The level of [Ca2+]i started to decline after washout .The Ca2+elevation induced by Ang II was significantly decreased by apocynin or TEMPOL .No significant differ-ence in the cell viability between control group and 5μmol/L Ang II treatment group was observed .CONCLUSION:ROS is involved in the regulation of [Ca2+]i induced by Ang II in the primarily cultured medullary neurons , suggesting a poten-tial intracellular signaling mechanism involved in the Ang II-mediated oxidant regulation of central neural control of blood pressure.

Objective To evaluate the treatment outcomes and toxicities of intensity-modulated radiation therapy (IMRT) combined with chemotherapy for children and adolescents with nasopharyngeal carcinoma.Methods Forty-three nasopharyngeal carcinoma patients less than 19 years old were recruited between April 2010 and April 2016.All patients were treated with IMRT (total dose 61.2-76 Gy) combined with cisplatin based chemotherapy.The Kaplan-Meier test was used to calculate overall survival (OS) and progression-free survival (PFS).The patient's clinical characteristics,side effects and longterm effects of treatment were retrospectively analyzed.Results Among 43 patients,there were 29 (67.4％) male and 14 (32.6％) female,and the median age was 14 years old (range,6-18 years).According to AJCC 7thstaging system,2 patients were in stage Ⅱ,26 in stage Ⅲ,7 in stage ⅣA and 8 in stage Ⅳ B.All patients were confirmed as non-keratinizing carcinoma.The positive rates of EB virus VCAIgA was 53.8％ (7/13),and Rta-IgG was 60.0％ (6/10) before treatment.The median radiation dose was 70 Gy (range,61.2-76 Gy) to the primary tumor.Thirty-three (76.7％) patients received neoadjuvant chemotherapy,with 20 (46.5％) and 36 (83.7％) patients treated by concurrent and adjuvant chemotherapy,respectively.With a median follow-up of 24 months (range,3-76 months),the 5-year OS and PFS ratios were 75.3％ and 64.7％,respectively.There were 5 patients (11.6％) occurred to bone metastasis within 2 years after treatment.Hypothyroidism was reported in 47.4％ (9/19) patients after IMRT.Conclusions Nasopharyngeal carcinoma in childhood and adolescence is mostly locally advanced diseases with poor differentiation.IMRT combined with chemotherapy produce a well treatment outcome with good tolerance in children and adolescents patients.The most common treatment failure bone metastasis.Radiation-induced hypothyroidism is common.

Objective To explore the impact of early repetitive painful procedures on subsequent pain behaviors and physiological indicators in full-term infants.Methods Sixty-two full-term neonates were enrolled (male 36 cases,female 26 cases) from the medical center of neonatal care at Nanjing Children's Hospital Affiliated to Nanjing Medical University from March to May in 2009.The data of all painful procedures were performed on those neonates and their responses to them were collected by using digital video recording were collected.The Neonatal Facial Coding System C(NFCS) and Neonatal Infant Pain Scale(NIPS) were used prospectively to evaluate the pain response to painful procedures.Results The average experience of pain caused by operation on newborn at the hospital was 56.5 times (12-249 times),and the daily average was 5.9 times (4-26 times).After they were exposed to more than 10 repetitive pain procedures,the full-term neonates showed the declining pain response assessed by NFCS and NIPS during the subsequent painful procedures (NFCS:P10=0.012,P20=0.015,P30=0.041;NIPS:P10=0.006,P20=0.015,P30=0.049),and the temporarily enhanced pain response was observed in the preparation phase of the subsequent painful procedures (NFCS:P2＜0.001,P3＜0.001,P4=0.004,P5=0.009;NIPS:P2＜0.001,P3＜0.001,P4=0.045,P5 =0.031).Bnt there was no difference in preparation phase.There was no alternation in latency and crying time after repetitiing painful procedures,but the proportion of crying neonates in the preparation phase of the painful procedures was increased(P=0.032).Conclusions After exposed to repetitive pain,full-term neonates expressed hypoalgesia during subsequent painful procedures,but the temporary irritation was enhanced during the following phases exposed to stress.

Hemodynamic situation is an important factor of recurrence of postoperative carotid artery aneurysm. In order to investigate the hemodynamic factors of postoperative carotid artery aneurysm affect carotid artery aneurysm recurrence, we established a 3D finite element carotid artery aneurysm for the preoperative and postoperative periods using the three-dimensional reconstruction techniques. And then we measured the hemodynamic factors of carotid artery aneurysm of preoperative and postoperative by the finite element method. The carotid artery aneurysm model has an accurate and realistic shape; the pressure of the recurrence of aneurysm was reduced significantly after surgery,wall shear stress increased significantly at residual neck, and blood flow velocity increased significantly, which will increase the risk of recurrence. The hemodynamic analysis provides a reference for development of aneurysm clinical treatment programs and prevention of recurrence.
Aneurysm ; pathology ; surgery ; Blood Flow Velocity ; Carotid Arteries ; pathology ; surgery ; Computer Simulation ; Hemodynamics ; Humans ; Stress, Mechanical

Objective:To analyze the characteristics of the molecular transmission network of newly-diagnosed human immunodeficiency virus type 1 (HIV-1) infected individuals in Shaoxing City, Zhejiang Province, and to provide evidence for epidemic trend and prevention.Methods:The plasma samples from 423 antiretroviral-naive HIV-1/acquired immunodeficiency syndrome patients from August 2018 to December 2019 were collected, and the pol gene fragments of HIV-1 from 375 samples were amplified by reverse transcription polymerase chain reaction (PCR) and nested PCR. The phylogenetic tree was constructed to analyze the molecular transmission network for subtypes and different gene distances by MEGA 6.0 software, HyPhy software and Cytoscape 3.7.2. Mutations on drug resistance was analyzed by online software tool of the HIV drug resistance database of Stanford University. Results:Eight subtypes were found in the 375 samples. Circulating recombinant form (CRF)07_BC(215/375, 57.33%) and CRF01_AE(103/375, 27.47%) were the major subtypes, followed by CRF85_BC, CRF55_01B, B, C, and CRF01_AE/B subtypes. One hundred and ninety-four individuals (51.73%) were connected to the transmission network at 1.50% genetic distance with 24 clusters. One hundred and twenty-nine individuals (34.40%) were connected to the transmission network at 0.75% genetic distance with 30 clusters, and 35 elderly patients were clustered in CL1.Forty-two cases had surveillance drug resistance mutation (SDRM), the prevalence of transmitted drug resistance was 11.20%(42/375). Thirty-eight cases had the drug mutations to non-nucleoside reverse transcriptase inhibitor, including K103 N(32/375, 8.53%), K103 S(4/375, 1.07%), Y188 L(1/375, 0.27%) and G190 A(1/375, 0.27%); four cases had the mutations to protease inhibitor, including M46 I(3/375, 0.80%) and V82 A(1/375, 0.27%). The cluster C2 sequences carried a high proportion of resistant mutations (94.29%, 33/35). Conclusions:HIV-1 subtypes in Shaoxing City are diverse and the CRF07_BC subtype spreads rapidly. The elderly patients with drug resistance genes in cluster CL1 at 0.75% gene distance need to be intervened immediately to prevent the drug resistance virus spread.

Objective: To investigate the molecular transmission network characteristics of HIV-1 among men who have sex with men (MSM) in Shaoxing City, Zhejiang Province, so as to provide insights into AIDS prevention and control among MSM. Methods: Newly reported HIV/AIDS cases among MSM in Shaoxing City were selected from January 2021 to September 2023. Plasma samples before the antiviral treatment were collected. The HIV-1 pol gene was amplified using reverse transcription PCR and nested PCR to construct phylogenetic trees for gene subtype analysis. The HIV-TRACE method was used to construct a molecular transmission network with a genetic distance of 1.5% to analyze clustering and the characteristics of cases within molecular clusters. Results: A total of 216 HIV/AIDS cases among MSM were included, and 179 qualified sequences were obtained. The predominant HIV-1 subtypes were CRF07_BC and CRF01_AE, with 95 and 66 cases, respectively. At 1.5% genetic distance, 20 molecular clusters were formed, with 61 nodes and 58 edges. A total of 61 sequences were connected to the transmission network (34.08%). HIV/AIDS cases among MSM from all the counties (cities, districts) in Shaoxing City were included in the network. There was the largest molecular cluster with ≥10 nodes, involving 12 cases from five counties (cities, districts), 3 medium-sized molecular clusters with 4 to 5 nodes, and the 16 small-sized molecular clusters with 2 or 3 nodes. Seven cases with high risk of transmission, each with ≥4 edges, were all CRF07_BC subtypes. Among them, two cases were from the large molecular cluster, and five cases were from the same molecular cluster composed of cases from Shengzhou City and Xinchang County. Conclusions The predominant HIV-1 subtypes among MSM in Shaoxing City were CRF07_BC and CRF01_AE. There was cross-regional HIV transmission, and potential transmission risk might exist in Shengzhou City and Xinchang County.

Objective To apply the MRI T2 signal intensity of hippocampal subfield in patients with amnestic mild cognitive impairment (aMCI) as early imageology. Methods From October, 2014 to August, 2015, 20 aMCI patients accepted cognitive training (training group), 20 aMCI patients accepted speech communication (speech group), and 20 age-and gender-matched healthy old people (control group) were scanned with MRI using FSE-T2 sequence. The margin of hippocampal subfields were outlined manually for each side to measure the T2 sig-nal intensity. The correlation between hippocampal T2 signal intensity and the scores of Mini-Mental State Examinatlon (MMSE) was ana-lyzed in the training group. Results Before treatment, T2 signal intensity in the bilateral hippocampal head was significantly high in the aM-CI patients compared with that in the control group (P<0.05). After treatment, T2 signal intensity in left hippocampal head decreased in the training group compared with that in the speech group (P<0.05), similar to the control group (P>0.05). There was negative correlation be-tween left hippocampal head's T2 signal intensity and the scores of MMSE in the training group before and after treatment (r=-0.61, r=-0.54, P<0.05). Conclusion The T2 signal intensity in left hippocampal head may respond to the cognitive function in patients with aMCI in the early stage, that could be used for diagnosis and evaluation in clinic.

Objective : To estimate the risk threshold of route index（RI）and mosq-ovitrap index（MOI）based on Breteau index（BI）,as supplements for dengue fever risk monitoring in specific habitats. Methods : Two towns and two streets were selected from nine towns（streets）in Jiashan County,and then one village（community）was selected from each of them as a Aedes albopictus monitoring site. The BI,RI and MOI were employed at the same time and area from April to October in the year 2018. Linear regression models were built with RI,MOI and BI to calculate the dengue risk threshold of RI and MOI according to BI. Results : The linear regression model of BI（X）and RI（Y）was Y=0.145+0.662X（P<0.05）,of BI（X）and MOI（Y）was Y=3.423+0.524X（P<0.05）. If BI=5（having risk of transmission of dengue fever）,then RI=3.455（95%CI：1.717-5.198）,MOI=6.043（95%CI：-0.327-12.417）. If BI=10（having risk of outbreak）,then RI=6.765（95%CI：5.018-8.518）,MOI=8.663（95%CI：2.260-15.071）. If BI=20（having risk of epidemic）,then RI=13.385（95%CI：11.326-15.453）,MOI=13.903（95%CI：6.352-21.461）. Conclusion The dengue fever risk threshold of RI estimated by BI had a narrow 95%CI and could be applied for dengue fever risk assessment,while the risk threshold of MOI had a wide 95%CI and the application value needed further study.

Objective:To seek survival-related genes in glioblastoma and establish a survival-gene signature for predicting prognoses of glioblastoma using public databases.Methods:Three independent glioma databases (GEO GSE53733,CGGA,TCGA) with whole genome expression data were included for analysis.Survival-related genes were obtained by comparing the long-term (＞36 months) and short-term (＜12 months) survivors in the database GSE53733.CGGA was used as the training set to develop the signature and TCGA was used asthe validation set.Cox regression analysis and linear risk score assessment were conducted to look for prognostic signatures with survival-related genes.Principal components analysis,gene set enrichment analysis (GSEA),gene ontology (GO) and protein-protein interaction (PPI) analysis were performed to explore distinct expression profiles between risk grouped glioblastoma.Results:We totally found 211 survival-related genes and developed a signature with 17 survivalrelated genes for prognosis of glioblastoma.Based on this signature,the low-risk group had longer survival time while the high-risk group had shorter survival time.Additionally,the expression profiles between the high-risk and low-risk glioblastoma were different.Functional annotations revealed that the genes enriched in the high-risk glioblastoma were involved in immune systems and processes of extracellular matrix (ECM).Conclusion:The novel survival-gene signature can predict high-risk glioblastoma with shorter survival time,enhance immunosuppressive features,and increased invasion preferences.

Objective:To investigate the clinical features, imaging features, gene diagnosis, treatment and prognosis of autosomal dominant lateral temporal epilepsy (ADLTE) with heterozygous RELN mutation.Methods:Clinical data of an ADLTE family caused by a heterozygous mutation in the RELN gene diagnosed in September 2019 at Qilu Hospital of Shandong University were collected. The clinical characteristics of ADLTE were analyzed, and literature review was conducted.Results:The male proband, 22 years old, was admitted with the clinical manifestations including seizures begun at temporal lobe, which specifically manifested as a sudden emergence of binaural hum, lasting for more than 10 seconds, and the symptoms can self-recover quickly. Half a month later, generalized tonic-clonic seizures attacked subsequently after a similar auditory aura. There were no abnormal findings in interictal electroencephalography (EEG) and magnetic resonance imaging (MRI). Following the family history, his father had similar auditory symptoms around the age of 20, and occasional secondarily generalized tonic-clonic seizures appeared. Antiepileptic drug can control better. The whole exome sequencing of three people in the family revealed that both the proband and his father had NM-005045: c.6068T>C heterozygous mutation in the RELN gene.Conclusions:ADLTE mostly occurs in juveniles or early adulthood. The main clinical manifestations are focal seizures with auditory auras, which can be followed by generalized tonic-clonic seizures. There are no abnormal findings in the interictal EEG and MRI. ADLTE is sensitive to drug treatment and has good clinical prognosis. The study identified a novel heterozygous mutation NM-005045: c.6068T>C in RELN gene, which is responsible for the development of ADLTE.