Objective: To reveal the molecular biological mechanism of a rare Dc-variant individual using PacBio third-generation sequencing technology. Methods: ABO and Rh blood type identification, DAT, unexpected antibody screening and D antigen enhancement test were conducted by serological testing. The absorption-elution test was used to detect the e antigen. RHCE gene typing was performed by PCR-SSP, and the 1-10 exons of RHCE were sequenced by Sanger sequencing. The full-length sequences of RHCE, RHD and RHAG were detected by PacBio third-generation sequencing technology. Results: Serological findings: Blood type O, Dc-phenotype, DAT negative, unexpected antibody screening negative; enhanced D antigen expression; no detection of e antigen in the absorption-elution test. PCR-SSP genotyping indicated the presence of only the RHCE c allele. Sanger sequencing results: Exons 5-9 of RHCE were deleted, exon 1 had a heterozygous mutation at c. 48G/C, and exon 2 had five heterozygous mutations at c. 150C/T, c. 178C/A, c. 201A/G, c. 203A/G and c. 307C/T. Third-generation sequencing results: RHCE genotype was RHCE 02N. 08/RHCE-D(5-9)-CE; RHD genotype was RHD 01/RHD 01; RHAG genotype was RHAG 01/RHAG 01 (c. 808G>A and c. 861G>A). Conclusion: This Dc-individual carries the allele RHCE 02N. 08 and the novel allele RHCE-D(5-9)-CE. The findings of this study provide data support and a theoretical basis for elucidating the molecular mechanisms underlying RhCE deficiency phenotypes.

Objective To discuss the application of the"rotating guidewire and correcting the filter recovery hook direction technique"("rotation-correction loop technique"for short),a technique invented by the authors in clinical practice,in the retrieval of complex inferior vena cava filter(IVCF),and to discuss its technical skills and advantages.Methods The clinical data of 417 patients carrying an IVCF,who were admitted to the Department of Vascular Surgery of Second Hospital of Shanxi Medical University of China to retrieve IVCF between January 2022 and December 2022,were retrospectively analyzed.Taking the time spent on the retrieval of IVCF and the intraoperative radiation dose as the evaluation indicators,the advantages and disadvantages of the standard filter retrieval technique,the"rotation-correction loop technique"and the other loop-assisted techniques were compared.Results Both the intraoperative radiation dose and the time spent on the retrieval of IVCF using"rotation-correction loop technique"were remarkably lower than those of other loop-assisted techniques(P＜0.000 1).Conclusion For the retrieval of complex IVCF,especially for the IVCF which is heavily tilted and/or its recovered hook is attached to the vascular wall,the use of"rotation-correction loop technique"can shorten the time spent on the the retrieval of IVCF and reduce the intraoperative radiation dose.This technique carries high safety and practicability,the device is simple and it can be manipulated by single physician,which is conducive to clinical application and promotion.(J Intervent Radiol,2024,33:289-294)

Background:When colonic adenoma involves diverticulum,inflammation in the diverticulum will increase the risk of adenomatous dysplasia.Therefore,patients with colonic diverticulum adenoma are at a higher risk of colon cancer,and the adenoma needs to be resected to avoid poor outcome and improve prognosis.Aims:To explore the efficacy and safety of endoscopic mucosal resection(EMR)in treatment of colonic diverticulum adenoma.Methods:Fourteen consecutive cases of colonic adenoma near or involving a diverticulum treated by EMR from Jun.2018 to Jan.2022 at the First People's Hospital of Hangzhou Lin'an District were collected,and their clinical characteristics and outcomes were analyzed.The primary outcome was complications,including bleeding,perforation,and electrocoagulation syndrome,while the secondary outcomes were the en bloc resection rate,complete resection rate and local recurrence rate.Results:Among the 14 patients with colonic diverticulum adenoma,13 were type A(near a diverticulum)while 1 was type B(involving a diverticulum).The diameter of the lesion was(0.76±0.25)cm,and the operation time was(19.67±5.33)minutes.The main histological type was tubular adenoma,and the pathological results was intraepithelial neoplasia in most of the cases.Delayed hemorrhage was observed in 1 patient(7.1%),and electrocoagulation syndrome in 1 patient(7.1%).No perforation event occurred.The en bloc resection rate and complete resection rate were 100%.Ten patients accepted reexamination of colonoscopy within 1 year after surgery,and no local recurrence was found.Conclusions:EMR is safe and effective for treatment of colonic diverticulum adenoma.However,patients using antiplatelet drugs and adenoma involving both appendiceal orifice and diverticulum should be alert to postoperative complications.

Objective:To investigate the utility of 68Ga-fibroblast activation protein (FAP) inhibitor (FAPI)-04 PET imaging in assessing the therapeutic response in pressure overload-induced heart failure. Methods:Rat models of pressure overload-induced heart failure were established by abdominal aortic constriction (AAC). Thirty rats were categorized into AAC group, 7 days reverse AAC (rAAC) group, and sham operation (sham) group ( n=10 per group) using completely random grouping method. All rats underwent 68Ga-FAPI-04 PET/CT imaging at 4 and 8 weeks after the ACC operation, while echocardiography, pathological examination, and immunohistochemical analysis were performed at 8 weeks postoperation. One-way analysis of variance, independent-sample t test and Pearson correlation analysis were used for data analysis. Results:68Ga-FAPI-04 PET/CT imaging showed that the target-to-background ratios of the heart and liver had significant differences among three groups both at 4 and 8 weeks postoperation ( F values: 2 547.12, 2 041.71, 462.65, 1 210.97, all P<0.001). Echocardiography revealed left ventricular ejection fraction (LVEF), left ventricular internal diameter at end-diastole (LVIDd), and left ventricular internal diameter at end-systole (LVIDs) in three groups at 8 weeks postoperation were significantly different ( F values: 118.92, 9.11, 10.63, all P<0.01). Rats were sacrificed at 8 weeks postoperation, and Masson staining showed that the fibrosis in the heart and liver of the rAAC group was significantly improved compared with that of the AAC group, and immunohistochemical analysis revealed significantly lower FAP levels in the heart and liver of the rAAC group compared with those of the AAC group ( t values: from -11.27 to -4.16, all P<0.01). FAPI uptake in the heart of the AAC group and rAAC group at 8 weeks postoperation were significantly positively correlated with FAPI uptake in the liver, LVIDd and LVIDs, FAPI uptake in the heart was significantly negatively correlated with LVEF, and FAPI uptake in the heart and liver were significantly positively correlated with fibrosis degree and FAP levels of corresponding organs ( r values: -0.89, -0.88, 0.72-0.97, all P<0.05). Conclusions:68Ga-FAPI-04 PET/CT can show the improvement process of cardio-liver fibrosis following the unloading of excessive pressure in heart failure. Myocardial FAPI uptake is closely related to the extent of heart failure improvement.

Objective:To investigate the etiology of hand, foot and mouth disease (HFMD) in Mianyang city in 2022, and to analyze the genetic characteristics of coxsackievirus A6.Methods:Pharyngeal swabs were collected from patients with HFMD in Mianyang city in 2022. Real-time fluorescence quantitative PCR was used to detect enteroviruses in the samples. Part of the VP1 gene in enterovirus-positive samples was amplified by nested PCR using enterovirus typing primers to further identify the viral types. The VP1 coding region of all CVA6-positive samples and the whole genome of some samples were amplified and sequenced by PCR. The endemic strain in Mianyang city was analyzed for phylogeny, gene homology, amino acid variation and genetic recombination.Results:A total of 151 pharyngeal swabs were collected, and 104 enterovirus-positive samples were detected by real-time fluorescence quantitative PCR, with an overall detection rate of 68.88% (104/151). The typing results showed that there were 77 cases of CVA6 infection, with a positive rate of 50.99% (77/151). The full-length VP1 genes of 77 CVA6 strains were amplified, sequenced, and successfully spliced, and phylogenetic analysis showed that all 77 strains were of the D3 genotype. There were multiple amino acid variant sites in the prevalent strains in Mianyang city compared with the reference strain. Twenty whole genome sequences were amplified, sequenced, and successfully spliced, and homology analysis showed that the nucleotide homology between the 20 positive sequences ranged from 97.0% to 99.9%. Phylogenetic tree and recombination analysis showed that no recombination occurred in the coding regions of the epidemic strains in this study.Conclusions:The predominant pathogen causing HFMD in Mianyang city in 2022 is CVA6 D3 subtype, which is consistent with the national epidemic in 2022.

Objective:To understand the composition of respiratory virus spectrum in hospitalized children with acute respiratory infections in Guilin City from 2021 to 2022, and to explore the epidemic characteristics of different respiratory viruses.Methods:Information of a total of 638 hospitalized children with acute respiratory infections (ARI) syndrome under the age of 14 years in Guilin city, Guangxi from September 2021 to October 2022 was collected as research data. The collected pharyngeal swabs from pediatric patients were subjected to real-time fluorescence reverse transcription polymerase chain reaction for nucleic acid testing, screening for 15 common respiratory viruses, and analyzing the virus spectrum characteristics of hospitalized pediatric patients with acute respiratory infections.Results:Among the 638 specimens, 365 were tested positive for at least one virus, with a positive rate of 57.21% (365/638). The detection rate for two or more viruses was 12.85% (82/638). There were 12 types of viruses detected, and the difference in virus detection rates among different seasons was statistically significant (P<0.002). The virus positive detection rate of samples from different age groups was the highest in the 0-2-year old group (40.66%), followed by the 3-5-year old group (34.80%) and the 6-year and older group (24.54%).Conclusions:During the period from September 2021 to October 2022, all 12 respiratory viruses were prevalent in Guilin area, with obvious summer peak as characteristics. It is suggested that corresponding prevention and control measures should be focused on pathogens that may cause acute respiratory infections in children during the local summer to prevent and control the spread and prevalence of respiratory infections.

Objective:To understand the prevalence and characteristics of Rhinovirus (HRV) infection in influenza-like Illness (ILIs) patients in Mianyang, Sichuan province, China.Methods:Throat swabs were collected from patients of ILIs in sentinel hospitals in Mianyang during 2021—2022. Real-time fluorescence quantitative PCR was used to detect 16 common pathogens. The VP4/VP2 coding region genes of HRV positive samples were amplified by nest PCR. The phylogeny, consistency and amino acid variation of different serotypes were analyzed and compared with reference sequences from GenBank database.Results:A total of 332 ILIs′ samples were collected with a virus detection rate of 58.73% (195/332) in Mianyang. Among them, 23 samples (23/332) were HRV-positive, and 18 VP4/VP2 sequences of HRV strains were successfully amplified. It was found that 13 HRV serotypes were detected in ILIs samples in Mianyang, which belonged to three genotypes, namely HRV-A (12 strains), HRV-B (5 strains) and HRV-C (1 strain).Conclusions:HRV was one of the pathogens of ILIs cases in Mianyang during 2021—2022, with HRV-A types as the dominant strains.

Objective:To establish a method via transfection of RNA to rescue coxsackievirus B3 B3 (CVB3).Methods:The efficiency of CVB3 genomic RNA extraction from three nucleic acid extraction reagents, Qiagen 57704, Qiagen 52904, and Trizol, and the transfection efficiency of viral RNA with two transfection reagents (Lipofectamine 2000 and Lipofectamine 3000) were compared. The efficiency of CVB3 rescue in Vero cells and HEK293T cells to determine the optimal conditions for rescuing CVB3.Results:The number of phagolysosomes for virus rescue by Qiagen 57704, Qiagen 52904, and Tizol kit extracted RNA was 13.33±1.53, 150±15.00, and 1.67±0.58, respectively, and there was a statistically significant difference in the efficiency of the three method of extracting CVB3 RNA to rescue the viral RNA ( F=268.920, P<0.001); The number of phage spots formed by Lipofectamine3000 and Lipofectamine2000 transfected RNA was 74.50±3.00 and 22.00±5.00, respectively, and the difference was statistically significant ( P<0.01); Qiagen 52904 reagent extracted CVB3 nucleic acid more efficiently than Qiagen 57704 and Trizol reagents; the transfection efficiency of transfection reagent Lipofectamine 3000 was 3 times more than than that of Lipofectamine 2000, and the efficiency of virus rescue of CVB3 in HEK293T cell culture was higher than that of HeLa and Vero cells, and the copy numbers of the three kinds of viruses rescuing the virus were 6.09×10 7±8.00×10 5, 5.18×10 3±6.17×10 2 and 0, the difference was statistically significant ( F=17 383.644, P<0.001), and it was also found that the efficiency of virus rescue could be improved by multiple elution when extracting RNA. Conclusions:In this study, we successfully established the method of transfecting RNA to rescue CVB3, which can effectively improve the efficiency of virus rescue by choosing Qiagen 52904 nucleic acid extraction kit, increasing the number of elution, using Lipofectamine 3000 transfection reagent, and transfection of HEK293T cells.

A subset of patients with non-alcoholic fatty liver disease(NAFLD)can progress to nonalcoholic steatohepatitis(NASH).When NASH reaches a fibrosis degree of F≥2 and a NAS score of≥4,this stage of NASH is referred to as fibrotic NASH,which is a key focus in clinical drug trials.Currently,liver biopsy is the gold standard for assessing the histological changes of the liver,but its clinical application is limited by its invasiveness,and therefore,it is of particular importance to develop noninvasive detection methods for fibrotic NASH.This article summarizes the recent research achievements in novel noninvasive diagnostic methods for fibrotic NASH and elaborates on these new diagnostic methods for predicting fibrotic NASH in terms of current status,challenges faced,and prospects for future development.

OBJECTIVE To analyze the pharmaceutical service process in a fracture patient complicated by fat embolism syndrome （FES） following postoperative fracture， aiming to provide a reference for clinical treatment and pharmaceutical service for similar patients. METHODS Clinical pharmacist participated in the entire treatment process of a patient with FES following postoperative fracture. Based on the patient’s clinical manifestations and test results， literature was reviewed to assist clinical physicians in formulating the therapeutic regimen of glucocorticoids. For the drug-related adverse reactions of renal function impairment and reduced platelet count that occurred during the treatment， suspicious drugs were analyzed and disposed of accordingly. RESULTS The clinical pharmacist recommended Hydrocortisone sodium succinate for injection （100 mg， q8 h， ivgtt， for about one week followed by a gradual dose reduction） for treating FES. The Vancomycin hydrochloride for injection used in this case was assessed as “very probably” associated with the adverse drug reactions of renal function impairment and thrombocytopenia. The clinical physician adopted the pharmacist’s medication recommendations， and the patient’s condition stabilized after treatment， with improvement in adverse reactions， and was discharged from the hospital. CONCLUSIONS The use of glucocorticoids in treating FES has a definite therapeutic efficacy. Clinical pharmacists should individualize the medication plan based on the patient’s pathological state and distinguish it from postoperative sepsis. Meanwhile， drug-induced adverse reactions in the kidney and blood system should be closely monitored.