Objective:To evaluate the feasibility, accuracy, effectiveness and safety of a novel manual placement of cervical 7 pedicle screws via the posterior approach of cervicothoracic junction.Methods:A retrospective case series study was conducted to analyze the 35 patients with injury to the lower cervical spine or cervicothoracic junction who had been treated by a novel manual placement of cervical 7 pedicle screws at Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University from March 2015 to July 2021. There were 16 males and 19 females, with an age of (52.7±13.2) years. The core of this placement was to determine the entry point of cervical 7 pedicle screws. After the intersection of the upper edge of the cervical 7 lamina and the medial edge of the superior articular process was recorded as point A while the intersection of the lateral edge of the inferior articular process and the lower edge of the transverse process as point B, the intersection of the outer and middle 1/3 of the AB line was taken as the screw entry point, with the screw placement angle perpendicular to the lamina line or slightly inclined from 30° to 40° to the head side and outward. The length, diameter and placement angle of the cervical 7 pedicle screws were recorded and compared postoperatively between the left and right sides to explore the feasibility of this novel manual placement. According to the Rampersaud method, the screw positions were graded 1 week and 6 months after operation to evaluate the accuracy of this manual placement. The visual analogue scale (VAS) and the Japanese Orthopaedic Association (JOA) score were compared between preoperation, 1 week and 6 months after operation to evaluate the effectiveness of this placement. The postoperative complications were counted to evaluate the safety of this method. Loosening, displacement and breakage of the screws were observed by CT scanning at 6 months after operation.Results:This case series was followed up for (9.8±1.7) months. There was no significant difference in the length, diameter or placement angle of the screws between the left and right sides ( P>0.05). A total of 66 cervical 7 pedicle screws were placed. There was no change in the screw position grading at 1 week or 6 months after surgery. Grade A was achieved in 64 screws, Grade B in 2 screws, and Grade C or D in none. The VAS scores before operation, 1 week and 6 months after operation were respectively 4.4±1.7, 3.8±1.0 and 1.1±1.1, and the JOA scores respectively 6.7±2.2, 13.2±1.5 and 15.3±1.2. The VAS and JOA scores at 1 week and 6 months after operation were significantly improved compared with the preoperative values ( P<0.05). The improvement rates in JOA at 1 week and 6 months after operation were 62.7%±13.3 % and 83.9%±11.6%, respectively. There were no complications related to the placement of cervical 7 pedicle screws; there was no wound hematoma or infection. No loosening, displacement or breakage of the screws was observed by the 6-month follow-up. Conclusion:The novel manual placement of cervical 7 pedicle screws via the posterior approach of cervicothoracic junction is feasible, accurate, effective and safe.

Objective:To explore the value of application and manipulation technique of neuroendoscope in microsurgical clipping of ruptured posterior communicating artery(PCoA)aneurysms via keyhole approaches.Methods:From January 2018 to December 2020, the clinical data of 52 patients who received microsurgical clipping for ruptured via keyhole approach were retrospectively analysed. Forty-one patients had the intraoperative endoscopic monitoring. The supraorbital keyhole approach or pterional keyhole approach was applied based on the characteristics of the aneurysms. According to the in-surgery requirement, a 30° rigid neuroendoscope was used before and/or after clipping. All patients entered postoperative follow-up in outpatient clinic and were evaluated with the modified Rankin Scale(mRS).Results:All 52 patients had 52 ruptured PCoA aneurysms. Eighteen of the patients were treated via supraorbital keyhole approach and 34 via pterion keyhole approach. Pre-and post-clipping endoscopic observation were carried out in 12 cases and 29 only with post-clipping endoscopic observation. Residual aneurysmal neck was detected in 3 patients. Missed clipping of perforators was found in 2 patients and followed by proper adjustment of clips. All patients received follow-up angiographic examinations. Total obliteration of the aneurysm and an intact of internal carotid artery and PCoA were found in 41 patients by the intraoperative endoscopic observation. Two residual aneurysmal neck were detected in 11 patients without intraoperative endoscopic observation. After 11 to 45 months of follow-up, all patients had good recovery(mRS 0-1).Conclusion:It is a safe and effective method with endoscopic observation during microsurgical clipping procedure for ruptured PCoA aneurysms via keyhole approaches. It can effectively make up for the insufficient visual angle of microscope, realise the anatomical relationship between the aneurysm and adjacent structures, and avoid residual aneurysmal neck and an iatrogenic injury to the parent artery and perforators.

ObjectiveTo screen out the mRNAs involved in the resistance of hepatoma cells to anlotinib using ceRNA microarray. MethodsHigh-dose shock combined with low-dose induction was used to culture hepatoma cells resistant to anlotinib, and CCK8 assay was used to verify the difference in the proliferation of drug-resistant hepatoma cells treated by anlotinib. The ceRNA microarray was used to screen out the differentially expressed genes between drug-resistant hepatoma cells and normal hepatoma cells, and real-time PCR was used to verify the differentially expressed genes detected by some microarrays. the independent samples t-test was used for comparison of continuous data between two groups, and the Kaplan-Meier method was used to analyze the overall survival of hepatoma cells samples, and the log-rank test was used to compare survival rates. Fisher’s exact test was used for chip screening. ResultsThere was a significant difference in gene expression between drug-resistant hepatoma cells and normal hepatoma cells, and 10 genes with the greatest difference were screened out for analysis by reducing the range. There were 4 genes associated with drug resistance and tumor growth, i.e., BIRC2, BIRC7, ABCC2, and MAPK8. There were significant reductions in the expression levels of BIRC2, ABCC2, and MAPK8 (P=0001 4, 0001 2, and 0.011 8), and there was a significant increase in the expression of BIRC7 (P＜0.001). The results of real-time PCR were consistent with those of microarray (t=10.74,32.65,18.34, and 2.80; P=0.000 4, 0.000 1, 0.000 1, and 0.044 8). The high expression of BIRC7 and the low expression of MAPK8 were associated with the significant reduction in survival time (P=0.022 0 and 0.005 6). ConclusionBIRC2, BIRC7, ABCC2, and MAPK8 are differentially expressed between anlotinib-resistant hepatoma cells and normal hepatoma cells and may be involved in the resistance of hepatoma cells to anlotinib.

Objective: To understand the epidemiological and etiological characteristics of enterovirus (EV)-associated diseases among children in Beijing from 2010 to 2016. Methods: This was a repeated cross-sectional study. The throat swabs were collected from children with probable EV-associated diseases at the Children' s Hospital Affiliated to Capital Institute of Pediatrics from 2010 to 2016. The samples were sent for pan-EV, enterovirus 71 (EV-A71) and coxsackievirus A16 (CV-A16) detection by real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) . The viral types of non-EV-A71 and non-CV-A16 EV-positive samples were identified using modified RT-PCR and sequencing with CV-A6, EV-A/B group and 5 'UTR universal primers. The constituent ratios of the prevalence of different EV types in different age and gender groups were compared. Results: Of the 2 703 throat swabs, 1 992 (73.7%) samples were positive for EV, including EV-A71 (19.1%, 516/2 703), CV-A16 (24.3%, 658/2 703), CV-A6 (22.2%, 600/2 703), CV-A10 (4.5%, 122/2 703) and other types of EV (3.5%, 95/2 703). There was 1 case of EV-A71 and CV-A16 co-infection. The positive detection rate of EV-A group (excluding EV-A71, CV-A16, CV-A6 and CV-A10) increased from 11.3% (7/62) to 95.2% (59/62) after using the modified VP1-specific primers and PCR amplification conditions. During the period between 2010 and 2012, CV-A16 and EV-A71 predominated in EV-positive samples. However, CV-A6 accounted for 60.7% (68/112) in 2013, much higher than CV-A16 (23.2%, 26/112) and EV-A71 (12.5%, 14/112). In 2014, EVs were mainly of CV-A16 and EV-A71, but CV-A6 was the predominant type in 2015 (68.2%, 232/340) and in 2016 (38.6%, 151/391). The epidemic season of EVs was mostly from April to August, but CV-A6 showed a small epidemic peak from October to November. The male-to-female ratio of EV-positive patients was 1.50∶1, and EV-associated diseases mostly occurred in children under 5 years of age. Younger children were more susceptible to CV-A6 than to EV-A71 and CV-A16. Conclusions From 2010 to 2016, there was a significant change in the spectrum of EVs in children with EV-associated diseases in Beijing. Since 2013, non-EV-A71 and non-CV-A16 increased, and CV-A6 gradually became one of the major pathogens of EV-associated diseases. The modified PCR primers and amplification conditions can effectively improve the reliability of test results.

Objective To prepare the phmaceutical reference materials of total lactones extract from ginkgo leaf for quantitative analysis.Methods A HPLC determination method was developed to investigate the uniformity and stability of the reference extract of ginkgo leaf total lactones using with bilobalide, ginkgolide A, ginkgolide B and ginkgolide C as the indexes.Three laboratories participated in the collaborative calibration test.Results The four components in reference extract had good uniformity and stability with RSD less than 2.0%;the marked values of the four components had been determined through statistical data analysis which provided by assigned traceability values.The values of bilobalide, ginkgolide A, ginkgolide B, and ginkgolide C were 39.54%, 29.03%, 15.96% and 11.69%, respectively.Conclusion The reference extract of ginkgo leaf total lactones can be prepared for quality control in future quantitative analysis.

Background and purpose:Breast cancer has the highest morbidity and mortality rate in women worldwide. Triple-negative breast cancer (TNBC) has no speciifc target and has low survival rate. Recent studies have veriifed BRD4 could promote tumor progression. This study aimed to detect the expression level of BRD4 in TNBC after treatment with gemcitabine, and to reveal the effect ofBRD4 silencing plus gemcitabine as a treatment for TNBC. Methods:The expression ofBRD4 in TNBC cell lines treated with gemcitabine was detected by reverse transcription PCR (RT-PCR) and Western blot. The effect of BRD4 silencing plus gemcitabine in TNBC was illustratedin vitro and in vivo.Results:The expression ofBRD4 in TNBC was signiifcantly increased after treatment with gemcitabine.In vitro,BRD4 knockdown signiifcantly lowered the IC50 value. The apoptotic rate of TNBC was signiifcantly increased in theBRD4 silencing plus gemcitabine group compared to the other. The growth rate of tumorin vivo was signiifcantly lowered in the BRD4 silencing plus gemcitabine group.Conclusion:BRD4 may play an important role in the drug resistance to gemcitabine in TNBC.BRD4 silencing plus gemcitabine may be a novel treatment strategy for TNBC.

OBJECTIVETo analyze the genotype, epidemic pattern and the characteristics of the disease of enteroviruses during the epidemic season of hand, foot and mouth disease (HMFD) in children from 2013 to 2014 in Beijing to provide the scientific evidence for prevention and treatment of HFMD.

METHODDuring April to September in 2013 and March to October in 2014, a total of 977 throat swabs were collected from children who visited the Children's Hospital Affiliated to Capital Institute of Pediatrics, including 147 from patients with HFMD in 2013, 343 with HFMD, 201 with atypical HFMD, 83 with herpangina, 25 with fever with convulsions, 64 fever with rash and 114 with rash in 2014. Enteroviruses universal type (EV), Enteroviruses type 71 (EV71) and Coxsackievirus group A 16 (CA16) were detected by real-time RT-PCR respectively. The nucleic acid of specimens which were identified with non-EV71, non-CA16 was tested by nested PCR and analyzed by VP1 sequencing. The detection rate and epidemic pattern of different genotypes of enterovirus were analyzed among different age groups and between 2013 and 2014.

RESULTOf 977 throat swabs, 80. 1% samples were detected positive for enteroviruses. The positive rates of CA16, EV71, CA6, CA10, CA4 and other EVs were 25. 6% (250/977), 18. 9% (185/977), 20. 0% (195/977), 5. 0% (49/977), 1.5% (15/977) and 9.1% (89/977), respectively. Twenty six of the 89 other EVs included CA2, CA5, CA8, CA9, CA12, CA14, CB2, CB5, E6, E9 and E25, each genotype of which was no more than 3. The nucleotide homologies shared among CA6, CA10 and CA4 strains between 2013 and 2014 were 94. 3% - 100%, 93. 8% - 99. 1% and 92.7% - 99. 8%, respectively. The positive rates of ≤1 year group were 71. 1% (106/149), which was lower than that of other age groups (all P <0. 05), but similar to that of >5 year group (χ2 =1. 181,P = 0. 277). In 2013, the positive rate of EV was 85. 7% (126/147) and the predominant genotype was CA6 54. 8% (69/126), followed by CA16 20. 6% (26/126) and EV71 11. 9% (15/126). In 2014, the positive rate of EV was 85. 4% (293/343) in the 343 children with HFMD, the predominant genotypes were CA16 with the positive rate of 42. 7% (125/293), EV71 with 38. 2% (112/293) and CA6 with only 11. 3% (33/293). In 2014, the positive rates of EV in 201 atypical HFMD, 83 herpangina, 25 fever with convulsions, 64 fever with rash and 114 rash were 83. 6% (168/201), 80. 7% (67/83), 76. 0% (19/25), 64. 1% (41/64) and 60. 5% (69/114), respectively. All genotypes of enteroviruses peaked mainly during May to August every year, but there were no obvious epidemiological pattern about each genotype.

CONCLUSIONCA6 became the main causative agent of HFMD in 2013, however, CA16 and EV71 predominated again in 2014 in Beijing. The clinical manifestations caused by CA6, CA10, CA4 and other genotype of enteroviruses differed from EV71 and CA16. Besides EV71 and CA16, more attention should be paid to CA6, CA10, CA4 and other type of enteroviruses.

Beijing ; epidemiology ; Child, Preschool ; Enterovirus A, Human ; classification ; Enterovirus Infections ; epidemiology ; virology ; Exanthema ; Fever ; Genotype ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Real-Time Polymerase Chain Reaction

Humans ; Liver Failure ; Lymphohistiocytosis, Hemophagocytic

BACKGROUNDAcute respiratory infection (ARI) is one of the most common infectious diseases in infants and young children globally. This study aimed to determine the virus profile in children with ARI presenting with different severities.

METHODSClinical specimens collected from children with ARI in Beijing from September 2010 to March 2011 were investigated for 18 respiratory viruses using an xTAG Respiratory Viral Panel Fast (RVP Fast) assay. The Pearson chi-square analysis was used to identify statistical significance.

RESULTSOf 270 cases from three groups of ARI patients, including Out-patients, In-patients and patients in the intensive care unit (ICU), viruses were detected in 176 (65.2%) specimens with the RVP Fast assay. The viral detection rate from the Out-patients group (50.0%) was significantly lower than that from the In-patients (71.1%) and ICU-patients (74.4%) groups. The virus distribution was different between the Out-patients group and the other hospitalized groups, while the virus detection rate and distribution characteristics were similar between the In-patients and ICU-patients groups. The co-infection rates of the Out-patients group, the In-patients group, and the ICU-patients group were 15.6%, 50.0% and 35.8%, respectively. In addition to respiratory syncytial virus (RSV) and adenovirus (ADV), human rhinovirus (HRV) was frequently detected from children with serious illnesses, followed by human metapneumovirus (hMPV), human bocavirus (HBoV) and coronaviruses. Parainfluenza virus 3 (PIV3) was detected in children with lower respiratory illness, but rarely from those with serious illnesses in the ICU-patient group.

CONCLUSIONIn addition to so-called common respiratory viruses, virus detection in children with ARI should include those thought to be uncommon respiratory viruses, especially when there are severe ARI-related clinical illnesses.

Antigens, Viral ; analysis ; Beijing ; Child ; Child, Preschool ; China ; DNA, Viral ; genetics ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza A virus ; genetics ; pathogenicity ; Male ; RNA, Viral ; genetics ; Respiratory Tract Infections ; diagnosis ; virology ; Rhinovirus ; genetics ; pathogenicity

OBJECTIVEHuman parechovirus (HPeV) is a single-stranded, positive sense RNA virus in the Parechovirus genus within the large family of Picornaviridae. As a possible new pathogen of neonatal sepsis, meningoencephalitis and other infections in young children, HPeV gets more and more attention. This study aimed to better understand the association of HPeV with central nervous system (CNS) infectious diseases and sepsis among hospitalized children in Beijing.

METHODA total of 577 cerebrospinal fluid (CSF) samples were retrospectively collected from 557 children suspected of CNS infections in 2012. Three hundred and fifty-one of them were male and 206 were female. HPeV was screened by reverse transcription-nested PCR (RT-nPCR) with the universal primers which target the highly conserved 5'UTR. The positive samples were genotyped by amplifying and sequencing for the VP3/VP1 junction region. The sequences were compared with the HPeV sequences from GenBank and performed phylogenetic analysis.Some samples other than CSF from HPeV positive children, including serum, nasopharyngeal aspirate and stool, were collected and carried out screening for HPeV.

RESULTWith the RT-nPCR by universal primers, HPeVs were detected in 18 out of 577 CSF samples obtained from 18 children with a positive rate of 3.1%. The ratio of male and female was 2: 1. There were no statistically significant differences on infection rate between boys (12/351, 3.4%) and girls (6/206, 2.9%). All of 18 positive CSF samples were negative for enterovirus, Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), and herpes simplex virus 1 and 2 (HSV).HPeVs from 10 positive CSF samples were genotyped successfully, consisting of 7 HPeV3 and 3 HPeV1. In addition, 2 of 8 serum samples were positive for HPeV3 and 1 of 2 stool samples were positive for HPeV 1. HPeVs were identified in CSF from children aged from 15 days to 14 years, in which 7 cases were infants younger than 3 months and 5 cases were infants from 3 months to one year. Three children older than the age of 9 years (9, 13 and 14 years) were positive for HPeV. Most of the children (6/8) infected with HPeV3 were younger than 3 months and were diagnosed as sepsis, while the rest of HPeV3 positive children were diagnosed as meningitis and bronchopneumonia. HPeV3 infection clustered in August, while HPeV1 in January.

CONCLUSIONHPeVs were associated with CNS infections and sepsis in hospitalized children in Beijing, especially in children younger than one year.HPeV3 was the predominant type identified in CSF.

Adolescent ; Age Distribution ; Central Nervous System Infections ; cerebrospinal fluid ; epidemiology ; virology ; Cerebrospinal Fluid ; virology ; Child ; Child, Hospitalized ; Child, Preschool ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Parechovirus ; classification ; genetics ; isolation & purification ; Picornaviridae Infections ; cerebrospinal fluid ; epidemiology ; virology ; RNA, Viral ; genetics ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons ; Sepsis ; cerebrospinal fluid ; epidemiology ; virology ; Sequence Analysis, DNA