1.Clinical study of CT-guided hematoma puncture drainage surgery combined with high-dose urokinase in treating hypertensive intracerebral hemorrhage
Xiaoxuan MA ; Weisheng RUAN ; Biao SHI ; Chunlong ZHAO ; Chao LYU
China Medical Equipment 2025;22(2):43-46,53
Objective:To observe the clinical effect of computed tomography(CT)-guided hematoma puncture drainage surgery combined with high-dose(≥50 000 U)urokinase in treating hypertensive intracerebral hemorrhage.Methods:The case data of a total of 90 patients with hypertensive intracerebral hemorrhage who underwent treatment in Zhangjiakou First Hospital from January 2022 to January 2023 were retrospectively selected as the study subjects.They were divided into a control group and an observation group using the average method,with 45 cases in each group.The conventional group received CT-guided hematoma puncture drainage surgery combined with conventional dose urokinase(20 000 U),while the observation group received CT-guided hematoma puncture drainage surgery combined with high-dose urokinase(50 000 U),with a 6-month follow-up for all patients.The changes of hematoma volume,length of hospital stay,levels of inflammatory indicators,the levels of hemodynamics,clinical efficacy,and incidence of postoperative complications after treatment between two groups were analyzed and compared.Results:After treatment,the hematoma volume at the 1st day[(34.64±5.53)mm3]and the 7th day[(34.64±5.53)mm3]post surgery of observation group were significantly lower than those[(44.57±6.85)mm3 and(16.64±3.24)mm3]of conventional group,and the differences of them between the two groups were statistically significant(t=7.567,10.570,P<0.05),respectively.The length of hospital stay of observation group[(16.86±6.63)d]was significantly lower than that of the control group[(23.47±10.34)d](t=3.610,P<0.05).The levels of matrix metalloproteinase-9(MMP-9),interleukin-6(IL-6),C-reactive protein(CRP),tumor necrosis factor-α(TNF-α)at 7th days post surgery of observation group were significantly lower than those of conventional group,and the differences of them between the two groups were statistically significant(t=5.826,10.202,7.661,2.724,P<0.05),respectively.The blood flow levels of middle cerebral artery(MCA),anterior cerebral artery(ACA),and posterior cerebral artery(PCA)after surgery in observation group were significantly higher than those in conventional group(t=2.833,3.329,3.973,P<0.05),respectively.The effective rate of treatment in observation group(95.56%)was significantly higher than that in conventional group(80.00%),and the difference was significant(x2=5.075,P<0.05).There was no significant difference in the incidence of postoperative complications between the two groups(P>0.05).Conclusion:CT-guided hematoma puncture drainage surgery combined with high-dose urokinase can significantly increase the dissolution rate of hematoma,and reduce the body's inflammatory response,and improve cerebral hemodynamic indicators,and effectively enhance clinical efficacy in treating hypertensive intracerebral hemorrhage,which have better treatment safety.
2.Application of autologous pedicled nasal mucosal flaps by "three-step" strategy in repairing of cerebrospinal fluid leakage following transsphenoidal pituitary adenoma surgery.
Shiyin QIN ; Zhuohui LIU ; Fengfeng JIA ; Biao RUAN ; Ruiqing LONG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(7):640-645
Objective:To assess the efficacy of "three-step" strategy for preparing autologous pedicled nasal mucosal flaps in repairing cerebrospinal fluid(CSF) leaks following transsphenoidal pituitary adenoma surgery. Methods:A retrospective study was conducted on the clinical data of 25 patients who developed CSF leaks after transsphenoidal pituitary adenoma surgery at the First Affiliated Hospital of Kunming Medical University between July 2012 and June 2022. Surgical repair was selected step by step using nasal septal mucosal flap with either the posterior septal artery or septal branch of the sphenopalatine artery as the pedicle, or a pedicled middle turbinate mucosal flap. All patients underwent ≥2-year endoscopic follow-up to assess flap viability and CSF leak recurrence. Results:The median postoperative hospital stay was 4 days. Five patients developed intracranial infections postoperatively. The follow-up period ranged from 2 to 12 years. Nasal endoscopic examinations showed good mucosal flap growth, with no recurrence of CSF leakage in any of the patients. Conclusion:High-flow cerebrospinal fluid(CSF) leaks following pituitary tumor surgery pose significant challenges for clinical repair. Based on intraoperative nasal septal mucosal preservation and the condition of sellar base CSF leakage, the "three-step" strategy for preparing autologous pedicled nasal mucosal flaps-utilizing posterior septal artery, ethmoidal artery-based, or pedicled middle turbinate mucosal flaps sequentially-is a safe and effective repair method.
Humans
;
Retrospective Studies
;
Pituitary Neoplasms/surgery*
;
Surgical Flaps
;
Nasal Mucosa/surgery*
;
Cerebrospinal Fluid Leak/surgery*
;
Adenoma/surgery*
;
Postoperative Complications/surgery*
;
Male
;
Female
;
Middle Aged
;
Adult
;
Aged
3.Clinical study of CT-guided hematoma puncture drainage surgery combined with high-dose urokinase in treating hypertensive intracerebral hemorrhage
Xiaoxuan MA ; Weisheng RUAN ; Biao SHI ; Chunlong ZHAO ; Chao LYU
China Medical Equipment 2025;22(2):43-46,53
Objective:To observe the clinical effect of computed tomography(CT)-guided hematoma puncture drainage surgery combined with high-dose(≥50 000 U)urokinase in treating hypertensive intracerebral hemorrhage.Methods:The case data of a total of 90 patients with hypertensive intracerebral hemorrhage who underwent treatment in Zhangjiakou First Hospital from January 2022 to January 2023 were retrospectively selected as the study subjects.They were divided into a control group and an observation group using the average method,with 45 cases in each group.The conventional group received CT-guided hematoma puncture drainage surgery combined with conventional dose urokinase(20 000 U),while the observation group received CT-guided hematoma puncture drainage surgery combined with high-dose urokinase(50 000 U),with a 6-month follow-up for all patients.The changes of hematoma volume,length of hospital stay,levels of inflammatory indicators,the levels of hemodynamics,clinical efficacy,and incidence of postoperative complications after treatment between two groups were analyzed and compared.Results:After treatment,the hematoma volume at the 1st day[(34.64±5.53)mm3]and the 7th day[(34.64±5.53)mm3]post surgery of observation group were significantly lower than those[(44.57±6.85)mm3 and(16.64±3.24)mm3]of conventional group,and the differences of them between the two groups were statistically significant(t=7.567,10.570,P<0.05),respectively.The length of hospital stay of observation group[(16.86±6.63)d]was significantly lower than that of the control group[(23.47±10.34)d](t=3.610,P<0.05).The levels of matrix metalloproteinase-9(MMP-9),interleukin-6(IL-6),C-reactive protein(CRP),tumor necrosis factor-α(TNF-α)at 7th days post surgery of observation group were significantly lower than those of conventional group,and the differences of them between the two groups were statistically significant(t=5.826,10.202,7.661,2.724,P<0.05),respectively.The blood flow levels of middle cerebral artery(MCA),anterior cerebral artery(ACA),and posterior cerebral artery(PCA)after surgery in observation group were significantly higher than those in conventional group(t=2.833,3.329,3.973,P<0.05),respectively.The effective rate of treatment in observation group(95.56%)was significantly higher than that in conventional group(80.00%),and the difference was significant(x2=5.075,P<0.05).There was no significant difference in the incidence of postoperative complications between the two groups(P>0.05).Conclusion:CT-guided hematoma puncture drainage surgery combined with high-dose urokinase can significantly increase the dissolution rate of hematoma,and reduce the body's inflammatory response,and improve cerebral hemodynamic indicators,and effectively enhance clinical efficacy in treating hypertensive intracerebral hemorrhage,which have better treatment safety.
4.Analysis of the Role of Pepsin in Vocal Cord Polyp and Vocal Cord Cancer
Jingyu GAO ; Renjing LUO ; Biao RUAN ; Chaowu JIANG ; Zhuohui LIU ; Ruiqing LONG ; Qiulin LIANG ; Ce ZHANG ; Lu SU ; Peng LI
Journal of Audiology and Speech Pathology 2024;32(1):21-24
Objective To investigate the expression of pepsin in vocal cord polyps and vocal cord cancer,and to compare the difference of pepsin expression.Methods From May 2020 to December 2021,27 patients with vocal cord polyp,27 patients with vocal cord cancer and 23 healthy volunteers were selected.RSI and RFS scoring scales were used for scoring,pepsin detection kit was used for saliva pepsin detection,and immunohistochemical methods were used to detect the expression of pepsin in vocal cord tissues of patients with vocal cord polyps and vocal cord cancer.Results The RSI score,RFS score and pepsin test kit results of vocal cord polyp group and vocal cord canc-er group were higher than those of non-vocal cord disease group,and the differences of the three indexes were statis-tically significant(P<0.05).RSI score,pepsin detection kit results and pepsin immunohistochemistry results of vocal cord polyp group showed no significant difference compared with vocal cord cancer group(P>0.05).The RFS score of vocal cord polyp group was significantly different from that of vocal cord cancer group(P<0.05).Conclusion Pepsin may be an important pathogenic factor of vocal cord polyp and vocal cord cancer,and play an im-portant role in the occurrence of these two diseases.The difference of pepsin expression in vocal cord polyp and vo-cal cord cancer suggests that pepsin may have different pathogenesis.
5.Inhibitory effect of miR-487a on M2-type polarization of gastric cancer tumor-associated macrophages by targeting TIA1
Yan QU ; Lin DAI ; Biao WANG ; Duji RUAN ; Yuchang ZHONG ; Xuefeng YANG
Journal of Jilin University(Medicine Edition) 2024;50(3):728-738
Objective:To discuss the inhibitory effect of microRNA-487a(miR-487a)on the M2 polarization of tumor-associated macrophages(TAMs)in gastric cancer,and to clarify its effect on the proliferation,invasion,and migration of the gastric cancer AGS cells.Methods:The TAMs from gastric cancer tissue and adjacent normal tissue macrophages(NTMs)from adjacent tissue of the primary gastric cancer patients were isolated and cultured.The human monocyte THP-1 cells were induced in vitro to differentiate into TAMs,and the differentiated M0,M1,and M2 macrophages were cultured for 24 h by conditioned medium(CM)to obtain the TAMs,M1-TAMs,and M2-TAMs respectively.The TAMs were transfected and then divided into blank group,inhibitor-NC group,miR-487a inhibitor group,miR-487a inhibitor+si-NC group,and miR-487a inhibitor+si-TIA1 group.The transfection efficiencies of the cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The M2-TAMs were co-cultured with the AGS cells,and divided into AGS group,AGS+inhibitor-NC group,AGS+miR-487a inhibitor group,AGS+miR-487a inhibitor+si-NC group,and AGS+miR-487a inhibitor+si-TIA1 group.RT-qPCR method was used to detect the expression levels of miR-487a and lymphocyte intracytoplasmic antigen-1(TIA1)mRNA in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue in various groups;Western blotting method was used to detect the expression level of TIA1 protein in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue and TAMs in various groups;flow cytometry was used to detect the levels of CD206 and CD163 in TAMs in various groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-10(IL-10),transforming growth factor-beta(TGF-β),vascular endothelial growth factor A(VEGF-A),and arginase-1(Arg-1)in culture supernatant of the TAMs cells;CCK-8 assay was used to detect the proliferative activity of the AGS cells in various groups;wound healing assay was used to detect the migration rates of the AGS cells in various groups;Transwell assay was used to detect the number of invasion AGS cells in various groups.Results:The RT-qPCR results shoued that compared with NTMs from adjacent tissue,the expression level of miR-487a in the TAMs from gastric cancer tissue was significantly increased(P<0.01)and the expression level of TIA1 mRNA was significantly decreased(P<0.01).Compared with TAMs,the expression level of miR-487a in M1-TAMs was significantly decreased(P<0.01),and the expression level of TIA1 mRNA was increased(P<0.01);the expression level of miR-487a in M2-TAMs was significantly increased(P<0.01),and the expression level of TIA1 mRNA was decreased(P<0.01).After transfection,compared with blank group and inhibitor-NC group,the expression level of miR-487a in the cells in miR-487a inhibitor group was significantly decreased(P<0.01),indicating successful transfection.The Western blotting results showed that compared with NTMs from adjacent normal tissue,the expression level of TIA1 protein in TAMs from gastric cancer tissue was decreased(P<0.01);compared with TAMs,the expression level of T1A1 protein in M1-TAMs was significantly increased(P<0.01),and the expression of TIA1 protein in M2-TAMs was significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the expression level of TIA1 protein in the cells in miR-487a inhibitor group was significantly increased(P<0.01);compared with miR-487a inhibitor+si-NC group,the expression level of TIA1 protein in the cells in miR-487a inhibitor+si-TIA1 group was significantly decreased(P<0.01).The flow cytometry results showed that compared with blank group and inhibitor-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased(P<0.01);compared with miR-487a inhibitor+si-NC group,the levels of CD206 and CD163 in the cells in miR-487a inhibitor+si-TIA1 group were significantly increased(P<0.01).The ELISA results showed that compared with blank group and inhibitor-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased(P<0.01);after co-transfection,compared with inhibitor-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased(P<0.01);compared with miR-487a inhibitor+si-NC group,the levels of IL-10,TGF-β,VEGF-A,and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor+si-TIA1 group were significantly increased(P<0.01).The CCK-8 assay results showed that compared with AGS group,the proliferation activity of the cells in AGS+inhibitor-NC group was significantly increased(P<0.01);compared with AGS+inhibitor-NC group,the proliferation activity of the cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the proliferation activity of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.01).The wound healing assay results showed that compared with AGS group,the migration rate of the cells in AGS+inhibitor-NC group was significantly(P<0.05);compared with AGS+inhibitor-NC group,the migration rate of the cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the migration rate of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.05).The Transwell assay results showed that compared with AGS group,the number of invasion AGS cells in AGS+inhibitor-NC group was significantly increased(P<0.01);compared with AGS+inhibitor-NC group,the number of invasion AGS cells in AGS+miR-487a inhibitor group was significantly decreased(P<0.01);compared with AGS+miR-487a inhibitor+si-NC group,the number of invasion AGS cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased(P<0.01).Conclusion:Silencing the miR-487a expression can inhibit the M2 polarization of the gastric cancer-associated macrophages by targeted upregulation of TIA1,and suppress the proliferation,migration,and invasion of the gastric cancer cells.
6.Overexpression of TIA1 gene in M2-type tumor-associated macrophages inhibited invasion and migration of gastric cancer cells by regulating the PI3K/AKT signaling pathway
Yuchang ZHONG ; Duji RUAN ; Lin DAI ; Biao WANG ; Yan QU ; Xuefeng YANG
Chinese Journal of Immunology 2024;40(8):1658-1664
Objective:To investigate the effect of overexpression of T cell restricted intracellular antigen 1(TIA1)gene in M2-type tumor-associated macrophages(M2-TAMs)on invasion and migration of gastric cancer cells and its mechanism.Methods:Primary TAMs were extracted from gastric cancer tissues and induced to differentiate into M2-TAMs using IL-4 and IL-13.The TIA1 overex-pression plasmid(oe-TIA1)and its empty vector were transfected into M2-TAMs.The expression levels of TIA1 mRNA and protein were detected by qRT-PCR and Western blot.The expression levels of CD206 and CD163 were detected by flow cytometry.The levels of IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant were detected by ELISA.The transfected M2-TAMs were co-cultured with gastric cancer BGC-823 cells by Transwell co-cultivation system,and PI3K agonist 740Y-P was used to intervene in parallel.The cell migration ability was detected by scratch assay.Transwell assay was used to detect cell invasion ability.Protein expression levels of PI3K,p-PI3K,AKT,p-AKT,MMP-2 and MMP-9 in the cells were detected by Western blot.Results:①Compared with primary TAMs,the levels of CD206 and CD163 expression in M2-TAMs cells and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture superna-tant were significantly increased(P<0.05),while the expression levels of TIA1 mRNA and protein were significantly decreased(P<0.05).Overexpression of TIA1 gene significantly decreased the expression levels of CD206 and CD163 in M2-TAMs and IL-10,TGF-β,VEGF-A and Arg-1 in cell culture supernatant(P<0.05).②Overexpression of TIA1 gene in M2-TAMs could significantly reduce the migration and invasion ability and the expression levels of p-PI3K/PI3K,p-AKT/AKT,MMP-2 and MMP-9 proteins in BGC-823 cells(P<0.05).③740Y-P could significantly reverse the inhibitory effects of overexpression of TIA1 gene in M2-TAMs on migration,inva-sion and PI3K/AKT signaling pathway of BGC-823 cells.Conclusion:Overexpression of TIA1 gene in M2-TAMs can affect the inva-sion and migration of gastric cancer cells by blocking the PI3K/AKT signaling pathway.
7.Expert consensus on the clinical application of long-acting cabotegravir and rilpivirine
Lijun SUN ; Hongxia WEI ; Haibo DING ; Ping MA ; Hui WANG ; Lijing WANG ; Chunmei WANG ; Min WANG ; Qian WANG ; Hai LONG ; Jinchuan SHI ; Wei LYU ; Biao ZHU ; Jun LIU ; An LIU ; Lianguo RUAN ; Zaicun LI ; Linghua LI ; Huiqin LI ; Shenghua HE ; Meiyin ZOU ; Yuxia SONG ; Renfang ZHANG ; Jian ZHANG ; Xinping YANG ; Yahong CHEN ; Yaokai CHEN ; Hongxin ZHAO ; Qingxia ZHAO ; Zhongsi HONG ; Feng QIAN ; Guangyong XU ; Huihuang HUANG ; Wei CAO ; Jianhua YU ; Juan JIN ; Lin CAI ; Fujie ZHANG
Chinese Journal of Clinical Infectious Diseases 2024;17(6):431-439
The long-acting cabotegravir and rilpivirine injection regimen(CAB+RPV regimen)is the first approved long-acting antiretroviral therapy(ART)for HIV in China,administered once every two months. This regimen provides an innovative alternative to daily oral ART,benefiting virologically suppressed patients. Several large clinical-studies have shown that the CAB+RPV regimen achieves comparable virologic suppression and safety to daily oral regimens,while significantly enhancing patient satisfaction. Based on international and domestic HIV/AIDs guidelines and clinical evidence,this consensus offers expert recommendations on patient selection,clinical management,and key communication strategies for healthcare providers to support the effective use of this regimen,aiming to improve quality of life for people living with HIV and accumulate domestic clinical experience with this advanced treatment approach.
8.Inhibitory Effect of Crocin on Pituitary Adenomas via IRF7/NF-κB Signaling Pathway
Zhuohui LIU ; Shiyin QIN ; Hexiang ZHAO ; Fengfeng JIA ; Biao RUAN ; Ruiqing LONG
Journal of Kunming Medical University 2024;45(12):19-27
Objective To explore the role and mechanism of crocin in pituitary adenoma(PA)through clinical samples and related molecular biology experiments of HP75 cells.Methods From June 2022 to May 2023,16 PA samples were collected from the Second Department of Neurology and Otolaryngology skull base surgery of the First Affiliated Hospital of Kunming Medical University.Three normal control samples were from the human anatomy of the Forensic College of Kunming Medical University.The expression of IRF7 mRNA in clinical samples was detected,and the proliferation,migration,invasion and apoptosis of HP75 cells were detected by knocking down the expression of IRF7;the expression of NF-κB was regulated by IRF7 in HP75 cells,and crocin regulated the growth of PA cells and its regulatory effect on IRF7/NF-κB signaling pathway.Results RT-qPCR and immunohisto-chemistry showed that compared with the normal control group,the expression of IRF7 mRNA in PA was significantly increased(P<0.001);the expression of IRF7 protein in si-IRF7 group was significantly decreased(P<0.001);CCK-8,Transwell and flow cytometry results showed that compared with the control group,knockdown of IRF7 significantly decreased the cell viability of HP75 cells(P<0.001),inhibited the migration and invasion(P<0.001),and promoted the apoptosis of HP75 cells(P<0.001).In addition,knockdown of IRF7 could inhibit the expression of p-NF-κB p65/NF-κB p65(P<0.001)and p-NF-κB p65/NF-κB p65(P<0.001).Overexpression of IRF7 partially reversed the effect of crocin(P<0.001)and restored the expression of p-NF-κB p65/NF-κB p65(P<0.01).Finally,the biological behavior of HP75 cells showed that compared with crocin group,overexpression of IRF7 could improve the cell viability of HP75 cells,promote their migration and invasion,and inhibit cell apoptosis(P<0.001).Conclusion Crocin treatment can inhibit the proliferation,migration and invasion of PA cells,promote cell apoptosis,and alleviate the development of PA.In the mechanism,IRF7 is significantly overexpressed in PA,and knockdown of IRF7 can inhibit the malignant growth of PA.Crocin can inhibit the proliferation,migration and invasion of PA cells,and promote apoptosis by inhibiting IRF7/NF-κB signaling pathway.
9.Expert consensus on the clinical application of long-acting cabotegravir and rilpivirine
Lijun SUN ; Hongxia WEI ; Haibo DING ; Ping MA ; Hui WANG ; Lijing WANG ; Chunmei WANG ; Min WANG ; Qian WANG ; Hai LONG ; Jinchuan SHI ; Wei LYU ; Biao ZHU ; Jun LIU ; An LIU ; Lianguo RUAN ; Zaicun LI ; Linghua LI ; Huiqin LI ; Shenghua HE ; Meiyin ZOU ; Yuxia SONG ; Renfang ZHANG ; Jian ZHANG ; Xinping YANG ; Yahong CHEN ; Yaokai CHEN ; Hongxin ZHAO ; Qingxia ZHAO ; Zhongsi HONG ; Feng QIAN ; Guangyong XU ; Huihuang HUANG ; Wei CAO ; Jianhua YU ; Juan JIN ; Lin CAI ; Fujie ZHANG
Chinese Journal of Clinical Infectious Diseases 2024;17(6):431-439
The long-acting cabotegravir and rilpivirine injection regimen(CAB+RPV regimen)is the first approved long-acting antiretroviral therapy(ART)for HIV in China,administered once every two months. This regimen provides an innovative alternative to daily oral ART,benefiting virologically suppressed patients. Several large clinical-studies have shown that the CAB+RPV regimen achieves comparable virologic suppression and safety to daily oral regimens,while significantly enhancing patient satisfaction. Based on international and domestic HIV/AIDs guidelines and clinical evidence,this consensus offers expert recommendations on patient selection,clinical management,and key communication strategies for healthcare providers to support the effective use of this regimen,aiming to improve quality of life for people living with HIV and accumulate domestic clinical experience with this advanced treatment approach.
10.Application of PCR technique in etiological diagnosis of children with enterovirus and herpesvirus encephalitis
Sai LI ; Liya MO ; Can LIU ; Suwu YI ; Yang RUAN ; Yunhua LIU ; Kuanpeng GUO ; Biao LIU ; Na LIU ; Liping LI
Chinese Journal of Laboratory Medicine 2021;44(4):323-327
Objective:To examine the infection of the enterovirus and human herpes virus in children with suspected encephalitis.Methods:A total number of 365 suspected encephalitis cases were included in this study from August 2017 to December 2019 in Hunan Children′s Hospital. The clinical samples, the cerebrospinal fluid (CSF), serum, sputum, stool and urine were collected and preserved at-80 ℃condition. The enterovirus (EV) and human herpesvirus (HHV) were examined by a one-step nested reverse transcription PCR(RT-PCR) and a real-time fluorescent quantitative PCR (qPCR), respectively. The positive rate of the two viruses in clinical specimens of children with suspected encephalitis was examined. Among all cases, 132 cases were diagnosed with EV encephalitis or HHV encephalitis.Results:the EV encephalitis were identified in 20.5% (75/365) children with suspected viral encephalitis; whereas HHV encephalitis infection was identified as 15.6% (57/365). Among the 75 cases of EV encephalitis, echo 6 was the main sub-type of these diseases 52.0% (39/75) and others were EV71 (30.7%, 23/75), echo11 (6.7%, 5/75), Coxsackie virus A group 6(CA6, 4.0%, 3/75), echo30 (1.3%, 1/75), echo9 (1.3%, 1/75), echo4 (1.3%, 1/75),Coxsackie virus B group 1(CB1, 1.3%, 1/75))and poliovirus(1.3%, 1/75).Human herpes virus type 6 (HHV6) was the most common pathogen in 57 cases of HHV encephalitis, accounting for 35.1% (20/57).The other pathogens were Cytomegalovirus (CMV, 31.6%, 18/57), Epstein-Barr virus (8.8%, 7/57), Herpes simplex virus 1 (HSV1, 10.5%, 6/57), HSV2 (8.8%, 5/57), and Varicella zoster virus (VZV, 1.8%, 1/57) .The virus in CSF detected significantly earlier than that in serum after onset. Virus could be detected in CSF 2-7 days after onset,but 7-26 days in serum. Conclusions:This study uses nested PCR and qPCR to detect pathogens in clinical specimens of children. This not only expands our understanding of the clinical examination and diagnosis of viral encephalitis in children, but also promotes the method of this study to benefit more children.

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