BACKGROUND:MicroRNAs (miRNA) through regulating specific target gene mRNA expression play important roles in different processes of diseases. OBJECTIVE:To study the interaction of miRNA-21 with its target gene TLR4 in Hela cels. METHODS:The candidate target gene of miRNA-21 was determined according to miRNA analysis databases. The constructed recombinant adenovirus vector carrying pri-miRNA-21 gene was used, which could package and amplify viruses to transfect Hela cels. Then, the expression of fluorescent proteins was detected. Forty-eight hours after transfection of miRNA-21 or control, extracted proteins were used for detection of TLR4 protein using western blot assay. RESULTS AND CONCLUSION:Recombinant adenoviruses pAd/pri-miRNA-21 and pAd/neg at 100 MOI could successfuly infect Hela cels. Bioinformatic analysis suggested several possible binding sites between miRNA-21 and TLR4. The experimental results showed that miRNA-21 down-regulated TLR4 at protein levels, indicating that miRNA-21 can interfere with the expression of TLR4 target gene.

Objective To investigate the feasibility and safety of day surgery holmium:YAG laser lithotripsy for urinary tract calculi.Methods A total of 294 cases of urinary tract calculi were treated with ureteroscopic holmium:YAG laser lithotripsy from October 2010 to September 2011.There were 147 cases for day surgery and 147 cases for inpatient surgery group.The time of waiting to be admitted,success rate,operative time,intraoperative and postoperative complications,hospitalization costs were analyzed in each group.Results The mean operative time of the day surgery group and the inpatient surgery group was 46.4± 16.5 and 52.1± 18.3 min.The difference in the mean operative time between the two groups was not statistically significant (P＞0.05).The stone free rate was not statistically significant between the day surgery group (95.9％) and the inpatient surgery group (92.5％,P＞0.05).The complication rate of the day surgery group was slightly lower than the inpatient surgery group (5.0％ VS 5.4％),but the differences between the two groups was not statistically significant (P＞0.05).The mean time of waiting to be admitted of the day surgery group was statistically shorter than the inpatient surgery group(2.5±1.3d VS 7.6±3.6 d,P＜0.05).Patients in the day surgery groups spent statistically less money than patients in the inpatient surgery group (5433.4 VS 8612.9,P＜0.05).Conclusions Day surgery for ureteroscopic holmium:YAG laser lithotripsy could be safe with lower costs and incidence of postoperative complications,faster postoperative recovery.Pre-operative adequately and selecting appropriate patients are helpful to reduce and control of intraoperative and postoperative complications.

Objective To investigate the correlativity between secretin-stimulated magnetic resonance cholangiopancreatography (sMRCP) findings and pathological severity in a swine chronic
pancreatitis (CP) model. Methods Thirty-nine swine were divided randomly into control group (n=12) and experimental group (n= 27). In experimental group, the main pancreatic duct (MPD) was incompletely ligated to establish the model of obstructive CP. In control group, laparotomy was performed but without ligating the MPD. At the 4th, 8th and 12th week after modeling, one third swine of each group were undergone a series of dynamic sMRCP scans before (0 min) and at 1, 3, 5, 7, 9, 11 min after administration of secretin (0.6 μg/kg). And the MPD diameter and duodenum filling (DF) degree were measured. All survivals were sacrificed to pathological examination including HE and Van Gieson staining for histopathological grading. According to pathological severity, swine were divided into normal group, mild CP group and moderate to severe CP group. MRI features and indexes, including baselined diameter (BD), end diameter (ED), maximum diameter (MD), the largest expansion rate (LER), time to peak (Tpeak) and end change rate of pancreatic duct and duodenal filling (DF) scores were measured. The relationships between pathological grading and sMPCP indexes were analysed. The comparison of sMRCP data among the 3 groups were used ariance analysis, χ2 test and U test. Correlations between sMPCP indexes and pathological severity were tested using Spearman rank correlation coefficients. The diagnostic efficiency of sMRCP indexes were evaluated by ROC method. Results (1) In experimental group, 22 CP models were established and 19 CP swine (mild CP, n= 8; moderate and severe CP, n=11) were performed sMRCP successfully. Eleven swine in normal group were obtained satisfactory MRCP images. (2) sMRCP results:BD of 3 groups were (1.56 ± 0.46),(2.95 ± 1.17),(7.41 ± 1.91) mm, respectively. ED were (1.49 ± 0.31),(2.96 ± 1.17) and (7.37±1.90) mm, respectively. MD were (2.39±0.43),(3.91±1.27) and (7.86±1.87)mm, respectively. The median of LER were 42.10%, 34.85% and 6.58%, respectively. The median of DF scores were 3, 3, 2, respectively. The differences of above indexes have statistically significance (P values were all<0.01). There were correlation between sMRCP indexes (BD, ED, MD, LER and DFscores) and pathological severity (r values were 0.89, 0.92, 0.90,-0.85,-0.66, respectively and P values were all<0.01). Tpeak and end change rate of pancreatic duct had no significant differences (P values were>0.05),and no correlation with pathological severity(P values were all>0.05).For differential diagnosis between normal and mild CP, the area under ROC of BD, ED, MD, LER and DFscores were 0.915, 0.977, 0.926, 0.778 and 0.472, respectively and differential diagnosis between mild CP and moderate to severe CP group, the area under ROC were 0.966,0.966,0.960,1.000 and 0.915, respectively. Conclusions sMRCP findings of CP have characteristics and could be used for in vivo evaluation on the CP pathologic grades.

Objective To investigate the correlativity between elasticity modulus and pathological severity in chronic pancreatitis (CP).Methods Twenty-one pigs were divided randomly into experimental group (n=18) and control group (n=3) using random number method.The main pancreatic duct (MPD) was incompletely ligated to establish the CP model.In control group, MPD was not ligated.The animals were killed in batches at 4th, 8th and 12th week after surgery.The pancreatic tissue was taken for elasticity modulus test and pathological examination, and the pigs were classified into control, mild, moderate and severe groups based on the severity of fibrosis.Cell density, fat infiltration and extracellular edema were observed and classified into mild and severe.The difference of elasticity modulus among different groups were compared by Variance analysis, the correlation between pancreatic fibrosis and elastic modulus was analyzed with Spearman correlation analysis, and ROC curve was used to evaluate its efficacy of diagnosing CP.Results Sixteen CP models were established successfully expected for 2 deaths (mild, n=7;moderate, n=2 and severe, n=7).All of the control group (n=3) showed normal pancreas.The elasticity modulus of control, mild and moderate to severe group were 0.4268±0.0566, 0.3203±0.0518 and 0.2235±0.0685, respectively.The difference between the groups was statistically significant (F=13.658,P<0.01), and the elastic modulus and pathological grade had a negative correlation (r=0.969, P<0.01).AUC of elasticity modulus for differentiating normal and mild CP was 1.000, the best critical value was 0.3807, and both the sensitivity and specificity of the diagnosis were 100%.AUC for differentiating mild and moderate to severe CP was 0.8730, the best critical value was 0.2646, and the sensitivity and specificity of the diagnosis were 85.7% and 77.8%, respectively.The pancreatic elasticity modulus of low parenchymal cell density group and high parenchymal cell density group were 0.1931±0.0373 and 0.3485±0.0655, respectively, which in the high cell density group was significantly higher than that in the low cell density group (t=-5.719, P<0.01).The elasticity modulus of negative infiltration or slight fatty infiltration group and severe fatty infiltration group were 0.3401±0.0697 and 0.1855±0.0344, respectively, which in the negative infiltration or slight infiltration group was significantly higher than that in severe infiltration group (t=5.102, P<0.01).The elasticity modulus of negative or mild cell edema group and moderate to severe cell edema group were 0.2760±0.0825 and 0.3024±0.1056, respectively;there was no statistically significant(t=-0.586, P >0.05).Conclusions The elasticity modulus can be used to detect the pathological changes of CP, and evaluate the CP pathologic grades.

OBJECTIVE: To screen differentially expressed genes and gene pathways in L02 cell line stably expressing hepatitis C virus (HCV) Ib type nonstructural protein 4B (NS4B) mediated by lentiviral system, and to provide a basis for further research of molecular biological mechanism of NS4B gene in chronic hepatitis C and hepatocarcinogenesis. METHODS: NS4B stably overexpressed L02 cell line and negative control stable L02 cell line, designated as L02-NS4B and L02-mkate2 respectively, were resurrected and amplified in vitro. The differentially expressed genes between L02-NS4B and L02-mkate2 were determined by gene expression microarray from Human Gene 1.0ST. The significant pathways of the differential genes were selected by the Fisher's exact test and χ2 test according to kyoto encyclopedia of genes and genomes (KEGG) database. The differential expression levels of 5 selected genes including protein kinase C delta binding protein (PRKCDBP), tumor protein p53 (TP53), v-akt murine thymoma viral oncogene homolog 1 (AKT1), baculoviral IAP repeat containing 3 (BIRC3) and B-cell lymphoma 2-like1 (BCL2L1) from cDNA microarray data were further verified by real-time quantitative polymerase chain reaction (real-time QPCR). RESULTS: Between L02-NS4B and L02-mkate2, the genes with flurescence intensity ratio >1.2 or <0.8 were considered as differentially expressed genes. A total of 2 682 differentially expressed genes in the known 28 869 human genes were detected in L02-NS4B, 1 446 genes were upregulated and 1 236 genes were downregulated. A total of 41 involved pathways of up-regulated differential genes were identified by KEGG database, mainly including apoptosis, extracellular matrix receptor interaction, cell cycle, pathways in cancer and Toll-like receptor signaling pathway; and 20 involved pathways of down-regulated differential genes were identified, mainly including pathways in cancer, Wnt signaling pathway and cell cycle pathway. Of the 5 upregulated genes selected from cDNA microarray data, 3 genes showed the same differential expression pattern by real-time QPCR as that shown in cDNA microarray data, namely AKT1, BIRC3 and BCL2L1. The confirmation rate of real-time QPCR was 60%. CONCLUSION HCVNS4B can up-regulate or down-regulate the expression of many genes in L02 cells, thus affecting multiple signaling pathways relevant to cell apoptosis, cell cycle and carcinogenesis.
Cell Cycle ; Cell Line ; Gene Expression Profiling ; Hepacivirus ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Transcriptional Activation ; Viral Nonstructural Proteins ; genetics

Objective:To investigate the clinical efficacy of trigeminal neuralgia treatment in elderly patients with percutaneous balloon compression assisted by a robotic surgical navigation and positioning planning system.Methods:Eleven elderly patients with trigeminal neuralgia admitted to the Department of Neurosurgery, Beijing Chaoyang Hospital, Capital Medical University between August 2021 and August 2022 were retrospectively analyzed.Three-dimensional time-of-flight magnetic resonance angiography, a three-dimensional fast spin-echo sequence with variable flip angles and thin-slice CT scans were performed before the procedure, and multimodal image fusion was performed by a robot workstation.The volume of Meckel's cave was calculated.Two paths were set up in the surgical planning: an actual puncture path, targeting the anterior inner quadrant of the internal foramen ovale, and a virtual path, used to plan the insertion depth of the balloon.The actual puncture path was completed under the guidance of the robotic arm adapter, and the virtual path for depth measurement was completed under lateral X-ray views of digital subtraction angiography.The balloon was placed at the predetermined depth and was inflated to form a pear shape to complete the procedure.The visual analogue scale(VAS)score was used to evaluate preoperative pain, and the Barrow Neurological Institute(BNI)facial numbness score was used to evaluate postoperative facial numbness.The patients were followed up for 1-12 months and the results were analyzed.Results:In all of the 11 patients, puncturing the foramen ovale and entry into the Meckel's cave were successfully conducted and the balloon was inflated to form a pear shape.Ten patients(90.9%)achieved complete remission immediately after the procedure, and 1 patient had delayed healing at 5 days after the procedure.There were no serious complications related to the procedure.There was no recurrence of pain during the follow-up, and the BNI numbness grading was between 2-3.The ratio between the inflated balloon volume and the preoperative volume of the Meckel's cave was approximately 1.7.Conclusions:Robot-assisted percutaneous balloon compression is safe and effective for the treatment of trigeminal neuralgia in elderly patients.

Objective ToinvestigatethefeasibilityofpredictingtheKi-67expressionusing MRandclinicalfeaturesofbreast cancer.Methods 52patientswith56invasivebreastcarcinomawereretrospectivelyinvestigated.Eachsubjectunderwentpresurgical breastMRI,whichincludeddynamiccontrastenhancementandDWI(b=0s/mm2,1000s/mm2).Clinicalcharacteristicsand MR findingswerecollected,includingage,ER ,PR ,HER2,tumorsize,location,number,spiculesign,lobulationsign,margin,TICandADCvalues. Ki-67expressionwasrecordedbyimmunohistochemicalstaining,whichwasdividedintohighexpression(≥20%)andlowexpression (<20%).Leastabsoluteshrinkageandselectionoperator(Lasso)wereperformedtoselectthefeaturesmostassociatedwithKi-67 expression.ThenomogramwasconstructedtopredictKi-67expression,aswellasAUCand95%confidenceinterval(CI)werecalculated.Results TICtypesandADCvaluesassociatedwithKi-67expressionwereidentifiedbyLassoregression.Thenomogram predictedthehigh andlow Ki-67expressionwithanAUCof0.827 (95% CI:0.713-0.940).TheKi-67expressionwasnegativelycorrelatedwiththe ADCvalues(r=-0.430,P=0.003).Ki-67expressionwassignificantlycorrelatedwithTICtype,andTICⅢwasmorelikelytohavehigher expressionofKi-67.Conclusion ThereisindividualandrespectivecorrelationbetweentheexpressionlevelofKi-67andTICtype, ADCvalue.TheexpressionlevelofKi-67canbepredictedtosomeextentbyADCvaluesandTICtypes.

Objective:To analyze the molecular epidemiological characteristics of the Corona virus disease 2019 (COVID-19) cases in Shijiazhuang, which can reveal the origin of the outbreak and provide a scientific basis for COVID-19 prevention and control.Methods:From January 2 to January 8, 2021, a total of 404 samples from 170 COVID-19 cases were collected from the Shijiazhuang Fifth Hospital. The consensus sequence of 2019 novel Coronavirus(2019-nCoV) was obtained through multiplex polymerase chain reaction-based sequencing. The sequences of 170 COVID-19 cases were analyzed by the PANGOLIN, and the data were statistically analyzed by T-test.Results:Among the 404 COVID-19 samples, a total of 356 samples obtained high quality genome sequences (>95%,100×sequencing depth). The whole genome sequences of 170 COVID-19 cases were obtained by eliminating repeated samples. All 170 sequences were recognized as lineage B1.1 using PANGOLIN. The number of single nucleotide polymorphism arrange from 18-22 and most of the single nucleotide polymorphism were synonymous variants. All of 170 genomes could be classified into 48 sub-groups and most of the genomes were classified into 2 sub-groups (66 and 31, respectively).Conclusions:All cases in this study are likely originated from one imported case. The viruses have spread in the community for a long time and have mutated during the community transmission.