Objective To assess early clinical safety and efficacy of transfemoral transcatheter aortic valve replacement (TF-TAVR) for pure aortic regurgitation (PAR). Methods The clinical data of PAR patients who underwent TAVR in Wuhan Asia Heart Hospital and Wuhan Asia General Hospital from January 2018 to October 2022 were retrospectively analyzed. Patients were divided into a TF-TAVR group and a transapical transcatheter aortic valve replacement (TA-TAVR) group. The clinical data of the patients were analyzed. Results A total of 54 patients were enrolled, including 34 males and 20 females with an average age of 74.43±6.87 years. The preoperative N-terminal pro-B-type natriuretic peptide level was lower [808.50 (143.50, 2 937.00) pg/mL vs. 2 245.00 (486.30, 7 177.50) pg/mL, P=0.015], and the left ventricular end-diastolic diameter (56.00±6.92 mm vs. 63.07±10.23 mm, P=0.005) and sinus junction diameter (32.47±4.41 mm vs. 37.65±8.08 mm, P=0.007) were smaller in the TF-TAVR group. There was no death in the two groups during the hospitalization. Only 1 new death within postoperative 1 month in the TF-TAVR group (cerebral hemorrhage). A total of 2 new deaths in the TF-TAVR group (1 patient of sudden cardiac death and 1 of multiple organ failure), and there was no death in the TA-TAVR group within postoperative 3 months. There was 1 new death in the TA-TAVR group (details unknown), and there was no death in the TF-TAVR group within postoperative 6 months. There was no statistical difference between the two groups in the all-cause mortality and the cumulative survival rate during the follow-up period (P>0.05). The incidence of high atrioventricular block was 36.0% in the TF-TAVR group and 10.3% in the TA-TAVR group (P=0.024). There were no significant differences between the two groups in the perivalvular leakage (≥moderate), valve in valve, a second valve implantation, valve migration, cerebrovascular events, major vascular complications, complete left bundle branch block, new permanent pacemaker implantation or transferring to surgery (P>0.05). However, the incidence rates of complete left bundle branch block and new permanent pacemaker implantation were higher in the TF-TAVR group, accounting for 56.0% and 40.0%, respectively. Conclusion TF-TAVR is a safe and feasible treatment for PAR patients, which is comparable to TA-TAVR in the early postoperative safety and efficacy.

Objective:To analyze the genetic characteristics of the complete genome of a strain of human respiratory syncytial virus (HRSV) in Qinghai province in 2024.Methods:A total of 300 samples were collected during 2024 influenza surveillance in Qinghai province sentinel hospitals from patients with fever accompanied by severe respiratory infection symptoms. We used real-time fluorescent quantitative reverse transcription polymerase chain reaction RT-PCR) method to screen out HRSV subtype B (HRSVB) positive specimens, whole genome sequencing was performed on positivespecimens meeting the requirements for the sequencing. After downloading the global representative HRSVB genotypes at GenBank database, sequence alignment was performed, related evolutionary tree was built and the calculation and analyses of genetic distance were done, analyses of HRSVB sequencing of sequence homology of nucleotides, amino acids and amino acid mutation were performed.Results:The first strain in Qinghai, China/qinghai/2024-03 had a complete sequence of 15 140 bp nucleotides, with HRSV′s all structural characteristics, and subtype HRSVA prototype strain Long strains of nucleotide the lowest homology was 80.0%, and subtype HRSVB prototype strain nucleotide homology was above 94.7%. The result indicated that the first strain in Qinghai belonged to HRSVB subtype. Genetic evolution shows China/qinghai/2024-03 and USA/WA-S23450/2021 (OR326803.1) and Germany/2021 (OR795235.1) all belong to a branch, they have the closest relationship. Phylogenetic analysis of G gene showed that the strain belonged to BA9 genotype of HRSVB subtype, and the hypervariable regions of the genome were SH and G genes.Conclusions:In this study, the complete genome sequence of HRSV China/qinghai/2024-03 was obtained for the first time, and the basic molecular structural characteristics were elucidated, which filled the gaps in the gene and amino acid data of HRSV in our province, and also provided a basis for HRSV epidemiology.

Objective     To explore the association between the preoperative systemic immune-inflammation index (SII) and prognosis in non-small cell lung cancer (NSCLC) patients. Methods     A comprehensive literature survey was performed on PubMed, Web of Science, EMbase, The Cochrane Library, Wanfang, and CNKI databases to search the related studies from inception to December 2021. The hazard ratio (HR) and 95% confidence interval (CI) were combined to evaluate the correlation of the preoperative SII with overall survival (OS), disease-free survival (DFS), and recurrence-free survival (RFS) in NSCLC patients. Results     A total of 11 studies involving 9 180 patients were eventually included. The combined analysis showed that high SII levels were significantly associated with worse OS (HR=1.61, 95%CI 1.36-1.90, P<0.001), DFS (HR=1.50, 95%CI 1.34-1.68, P<0.001), and RFS (HR=1.17, 95%CI 1.04-1.33, P<0.001). Subgroup analyses also further verified the above results. Conclusion     Preoperative SII is a powerful prognostic biomarker for predicting outcome in patients with operable NSCLC and contribute to prognosis evaluation and treatment strategy formulation. However, more well-designed and prospective studies are warranted to verify our findings.

OBJECTIVE: To investigate iron accumulation level over the whole brain and explore the possible neuromechanism of medication-overuse headache (MOH) using quantitative susceptibility mapping (QSM). METHODS: Thirty-seven MOH patients and 27 normal control subjects were enrolled in the study for examinations with both a multiecho gradient echo magnetic resonance (MR) sequence and brain high resolution structural imaging. A voxel-based analysis was performed to detect the brain regions with altered iron deposition, and the quantitative susceptibility mapping values of the positive brain regions were extracted. Correlation analysis was performed between the susceptibility values and the clinical variables of the patients. RESULTS: In patients with MOH, increased susceptibility values were found mainly in the bilateral substantia nigra (SN) (MNI coordinate: 8, -18, -14; -6, -16, -14) as compared with the normal control subjects (P < 0.001), but these alterations in iron deposition were not significantly correlated with the clinical variables of the patients (P > 0.05). The susceptibility value in the left SN had an area under curve (AUC) of 0.734, and at the cut-off value of 0.077, its diagnostic sensitivity was 72.97% and its specificity was 70.37% for distinguishing MOH from normal controls; The susceptibility value in the right SN had an AUC of 0.699 with a diagnostic sensitivity of 72.97% and a specificity of 62.96% at the cut-off value of 0.084. CONCLUSION Increased iron deposition occurs in the bilateral SN of MOH patients, which provides a new insight into the mechanism of mesocorticolimbic dopamine system dysfunction in MOH. QSM technique can be used as a non-invasive means for quantitative analysis of brain iron deposition in migraine neuroimaging.
Humans ; Brain ; Substantia Nigra ; Magnetic Resonance Imaging/methods* ; Headache Disorders, Secondary ; Headache ; Iron ; Brain Mapping/methods*

Objective:Based on targeted amplicon technology combined with high-throughput sequencing technology and bioinformatic analysis technology, to understand the characteristics of the whole genome of the monkeypox virus and its variation, and to construct a method for the analysis of monkeypox virus variation and molecular traceability of the case in Qinghai province, and to provide technical support for the prevention and control of monkeypox epidemic in the future.Methods:The extracted viral DNA was used as a template, and the genome of monkeypox virus was specifically amplified by Ion AmpliSeq Monkeypox Panel with the number of amplicons 1 609 and the length of 125 bp-275 bp, and the sequencing library was constructed by Ion AmpliSeq Library Kit Plus, and sequenced by Ion Torrent GeneStudio S5. The sequencing library was constructed by Ion AmpliSeq Library Kit Plus, and the monkeypox virus genome was sequenced using Ion Torrent GeneStudio S5 sequencer. Monkeypox virus was analyzed for genomic profiling and mutation site analysis using the online analysis tool Nextclade. The genomic sequence of the case virus in this study was compared with some sequences in the GIASID monkeypox virus database and a phylogenetic tree was constructed to analyze the potential origin of the case virus.Results:The Ct values of monkeypox virus genes in the rash swab and oropharyngeal swab samples were 32.13 and 36.91, respectively. The rash swab sample had a reads number match of 99.99% and a genome coverage of 99.45% after whole-genome sequencing of monkeypox virus, and the sequences belonged to the IIb (West African branch) B. 1.3 type. The analysis of nucleotide mutation sites and phylogenetic tree showed that the sequences were in the same branch with four monkeypox virus genome sequences recently submitted by China and Japan in the GISAID monkeypox virus database, and had the closest evolutionary relationship with the sequence EPI_ISL_18059184 (sampled on 2023-07-03) submitted by Yunnan, China, which shared 82 single-nucleotide mutation sites, among which the sequence from Yunnan was only present in all of the shared 82 single-nucleotide mutation sites. The sequence in this study has 2 additional nucleotide mutation sites on top of the shared 82 single nucleotide mutation sites. The sequence submitted by Japan, EPI_ISL_17692269 (sampled on 2023-04-28), is more closely related in evolution, sharing 78 single nucleotide mutation sites, with 7 single nucleotide mutation site differences, and the Japanese sequence shares 78 single nucleotide mutation sites. The Japanese sequence shared 78 mutation sites with one additional nucleotide mutation site (G57786A), while the present sequence had six additional nucleotide mutation sites (G13563A, C21062T, G101241A, C142797T, G152866A, T169721A).Conclusions:The whole genome sequence of monkeypox virus of 197 084 bp was successfully obtained from a sample with low viral load, and the average. We constructed a method for sequencing and analyzing the whole genome of monkeypox virus.

The axon initial segment (AIS) is a specialized structure that controls neuronal excitability via action potential (AP) generation. Currently, AIS plasticity with regard to changes in length and location in response to neural activity has been extensively investigated, but how AIS diameter is regulated remains elusive. Here we report that COUP-TFI (chicken ovalbumin upstream promotor-transcription factor 1) is an essential regulator of AIS diameter in both developing and adult mouse neocortex. Either embryonic or adult ablation of COUP-TFI results in reduced AIS diameter and impaired AP generation. Although COUP-TFI ablations in sparse single neurons and in populations of neurons have similar impacts on AIS diameter and AP generation, they strengthen and weaken, respectively, the receiving spontaneous network in mutant neurons. In contrast, overexpression of COUP-TFI in sparse single neurons increases the AIS diameter and facilitates AP generation, but decreases the receiving spontaneous network. Our findings demonstrate that COUP-TFI is indispensable for both the expansion and maintenance of AIS diameter and that AIS diameter fine-tunes action potential generation and synaptic inputs in mammalian cortical neurons.
Action Potentials ; Animals ; Axon Initial Segment ; COUP Transcription Factor I ; DNA-Binding Proteins/physiology* ; Mammals ; Mice ; Transcription Factors

Overlap extension PCR is a common method for site-directed mutagenesis. As objective gene sequence growing longer, it is often difficult to obtain the target product in the second round of PCR, and it is highly possible to introduce unexpected mutations into a long gene fragment by PCR. To circumvent these problems, we can only amplify a small gene fragment which contain the target mutation by overlap extension PCR, and then ligate it with vector to get target plasmid. If the restriction site at the end of the amplified fragment was not a single one on plasmid vector, double fragments ligation method could be used to construct target plasmid. Partial amplification, combined with double fragments ligation, could solve lots of problems in long gene mutagenesis. Taking retinoblastoma gene 1 S780E mutagenesis as an example, it is difficult to amplify whole retinoblastoma gene 1 by overlap extension PCR because of long fragment interfering the overlapping extension of second round PCR. However, it is relatively easy to amplify the F3 (1 968-2 787) fragment which contains target mutation S780E. There is a Nhe I site which can be used for ligation on 5' end of F3 fragment, but another Nhe I site on the plasmid restrained from doing so directly. In order to circumvent this obstacle, we ligated F3 fragment, combining with F2 (900-1 968) fragment which was digested from wild type plasmid, with the vector which contain F1 (1-900) fragment of the gene. That double fragments ligated with one vector at the same time, though less efficient, can recombine into a complete plasmid. The sequences of the two selected recombinant plasmids were consistent with the target mutation, which verified the feasibility of this scheme. As an improvement of overlap extension PCR, partial amplification and double fragments ligation methods could provide solutions for site directed mutagenesis of many long genes.
Base Sequence ; Cloning, Molecular ; Genetic Vectors ; genetics ; Mutagenesis, Site-Directed ; methods ; Nucleic Acid Amplification Techniques ; Plasmids ; Polymerase Chain Reaction

（正）Luketich 等[1]2000 年首次报道了胸腹腔镜联合食管癌微创手术（minimally invasive esophagectomy， MIE）病例。随着近 10 多年的发展，食管癌微创外科已经进入一个技术成熟的阶段，能使广大食管癌患者受益。2009 年美国国立综合癌症网络（NCCN）临床指南[2]已将 MIE 列为标准食管癌术式之一。

Objective To investigate the efficacy of calf self weight traction reduction combined with locking plate fixation for the treatment of intertrochanteric fractures in the elderly.Methods A retrospective case series study was conducted on the clinical records of 174 elderly patients with the modified Evans Ⅰ-Ⅲ types of fresh intertrochanteric fractures treated with locking plate from January 2012 to December 2015.According to treatment method,the patients were assigned to traction bed reduction with locking plate fixation (Group A,62 patients) and calf self weight traction reduction with locking plate fixation (Group B,112 patients).Group A comprised 32 males and 30 females,with age range of 65-91 years [(72.47 ± 6.35) years].Group B comprised 68 males and 44 females,with age range of 65-95 years [(73.23 ± 6.05) years].The time of reduction,operation time,incision length,intraoperative blood loss,frequency of fluoroscopy,postoperative drainage volume,hospital stay,postoperative weightbearing standing time or walking time (ambulation time),surgical complications,and fracture healing were recorded.Harris and modified Barthel index score in Chinese (MBI-C) were used to evaluate the functional recovery of hip joint.Results All patients were followed up for 5-61 months (mean,15 months),and noted with fracture healing.The time of reduction in Group A was (13.27 ± 3.03) minutes,longer than that in Group B (0 minute) (P ＜0.05).The operation time in Group A was (63.63 ± 13.90)minutes,longer than that in Group B [(59.62 ± 8.38) minutes] (P ＜ 0.05).Fluoroscopy in Group A was (5.35 ± 2.36) times,more than (4.28 ± 3.11) times in Group B (P ＜ 0.05).There were no significant differences in the incision length,intraoperative blood loss,postoperative drainage volume,ambulation time,fracture healing time,Harris score,and MBI-C index between the two groups (P ＞0.05).There were no significant differences in the postoperative complications such as deep venous thrombosis,pulmonary infection,incisional infection,urinary tract infection,delirium,bed sores,cardiac insufficiency,electrolyte disturbance,and postoperative plate rupture between the two groups (P＞0.05).The incidence of deep vein thrombosis was 9.7％ (6/62) in Group A,and 4.5％ (5/112) in Group B (P ＞0.05).No screw fracture,nail and plate combination failure,bone nonunion,or screw cut out of the femoral head were observed in both groups.Conclusions For the modified Evans Ⅰ-Ⅲ types of intertrochanteric fractures,both traction bed reduction and calf weight reduction with locking plate have equivalent efficacy.However,the latter method has advantages of shorter reduction and operation time and less intraoperative X-ray exposure,and hence is worthy of clinical application.

Objective To systematically evaluate the postoperative complications following open reduction and internal fixation combined with vascularized bone flap graft versus only reduction and internal fixation for femoral neck fractures.Methods CNKI,Wan Fang Chinese database,Pubmed,EMBASE and Google Scholar English database were searched for the randomized controlled trials from January 1,2000 through October 31,2017 which compared open reduction and internal fixation combined with vascularized bone flap graft (combined surgery group) with only reduction and internal fixation (simple surgery group) for femoral neck fractures.The data concerning postoperative nonunion and avascular necrosis of the femoral head were extracted.The 2 surgical treatments of the patients with femoral neck fracture were compared in terms of the 2 complications.Statistical analyses were conducted using software Stata 12 by relative risk (RR) and corresponding 95％ confidence intervals (95％ CI).Results According to our inclusion and exclusion criteria,a total of 23 studies were included,involving 2,162 cases (1,048 cases receiving combined surgery and 1,114 cases simple surgery).The Meta analyses showed that the fracture nonunion rate for the combined surgery group was significantly lower than that for the simple surgery group [RR =0.27,95％ CI(0.19,0.38),P ＜ 0.001] and the rate of avascular necrosis was also significantly lower for the former than for the latter [RR =0.31,95 ％ CI(0.24,0.42),P ＜ 0.001].Conclusion In the treatment of femoral neck fractures,open reduction and internal fixation combined with vascularied bone graft may lead to lower rates of nonunion and avascular necrosis of the femoral head than simple open reduction and internal fixation.