Objective To explore the efficacy and safety of ultrasound-guided radiofrequency ablation combined with foam sclerotherapy in the treatment of primary small saphenous vein varicosities.Methods A total of 46 patients with primary small saphenous vein varicosities in our department from November 2021 to November 2022 were retrospectively analyzed.Depending on the patient's choice of surgical method,they underwent radiofrequency ablation combined with sclerotherapy(observation group,n=23)or high ligation and stripping of the saphenous vein(control group,n=23).The surgical time,intraoperative blood loss,length of hospital stay,postoperative Venous Clinical Severity Score(VCSS),incidence of postoperative complications,treatment satisfaction,and recurrence were compared between the two groups.Results Compared with the control group,the observation group had shorter surgical time[(56.1±6.0)min vs.(81.7±10.6)min,t=-10.128,P=0.000],less intraoperative blood loss[(27.0±4.1)ml vs.(41.4±4.8)ml,t=-11.016,P=0.000],shorter hospitalization time[(2.0±0.7)dvs.(5.6±0.9)d,t=-14.319,P=0.000],lower VCSS scores at 3,6,and 12 months after surgery[(7.4±2.0)points vs.(8.9±2.5)points,t=-2.165,P=0.036;(5.3±1.8)points vs.(6.5±2.2)points,t=-2.149,P=0.037;(2.6±1.3)points vs.(4.0±1.8)points,t=-2.912,P=0.006],higher satisfaction[satisfied,somewhat satisfied,dissatisfied in 18,4,and 1 cases in the observation group and 8,10,and 5 cases in the control group,respectively,Z=-2.967,P=0.003].There were no statistically significant differences in the complication incidence and recurrence rate between the two groups(P＞0.05).Conclusion Ultrasound-guided radiofrequency ablation combined with foam sclerotherapy in the treatment of primary small saphenous vein varicosities is safe and effective.

  Objective  To summarize the clinical features of a family with Basan syndrome and to analyze mutation of the SMARCAD1 gene.  Methods  The Basan family was diagnosed at Dermatology Hospital, Southern Medical University in 2022. Backgroud data was collected, and clinical and genetic characteristics were analyzed. Meanwhile, a retrospective analysis of features and associated genetic mutations reported in all patients with Basan syndrome was conducted.  Results  A total of 18 patients with Basan syndrome were identified, including 9 males and 9 females. All 18 patients had no fingerprints at birth (18/18, 100%), and some patients had knuckle pads, palmoplantar hyperkeratosis, nail atrophy, nail separation, and longitudinal nail ridges. Symptoms vary in severity. At the same time, it was found that c.-10+1G > T (as well as c.378+1G > T)mutations appeared on the intron 1 of the SMARCAD1 (NM_020159.5) gene in 7 patients, resulting in abnormal splicing.  Conclusions  This article provides help for the early diagnosis of Basan syndrome and helps to improve the diagnosis and differentiation level of clinicians.

Objective:To confirm the cross-provincial long-distance transmission events of Coronavirus Disease 2019 (COVID-19) by lorry drivers, the origin of infections of the cases and the transmission routes were tracked.Methods:Nasopharyngeal swab specimens from five lorry driver cases of COVID-19, found in Zhangzhou city in March, 2022 when the local outbreaks occurred in adjacent Quanzhou city, Fujian province, were collected to perform 2019 novel coronavirus (2019-nCoV) targeted genome amplification and followed by next-generation sequencing. The sequences were submitted to online 2019-nCoV analysis platforms to classify the type of variant and mutation sites. Phylogenetic tree for the viruses were constructed by phylogenetic analysis software. Combined with the epidemiological investigation, the origin of infections of the cases and the transmission routes were deduced.Results:Five complete genome sequences, with 29 770-29 839 bp in length and 99.53% average genomic coverage, of 2019-nCoV were successfully obtained. The viruses were all Omicron variants and further divided into three different subclades of BA.2. Of the five strains of 2019-nCoV, three were highly similar to the viruses of two distinct lineages co-circulated in Quanzhou city during the period of local outbreak of COVID-19, respectively. Phylogenetic analysis also revealed that the viruses from three infected lorry drivers were highly homologous to that from local outbreaks in Quanzhou city. The viruses from the rest two cases had seven to fourteen nucleotide mutations (corresponding to 5-7 amino acid substitutions) when compared with the viruses in local outbreaks in Quanzhou city, which excluded the involvement of the two cases into the transmission chains of local outbreaks. Combined with the field epidemiological investigations, the result revealed that the origin of infection of 2019-nCoV of the two sporadic lorry driver cases was outside of Fujian province.Conclusions:With the aid of high-throughput sequencing and bioinformatics technology combined with field epidemiological investigations, we speculated in this study that at least three origins of infection of 2019-nCoV in five lorry driver cases and cross-provincial long-distance transmission via two sporadic cases infected outside Fujian province when they returned.

Objective:To improve the efficiency and success rate of human adenovirus 3 (HAdV-3) whole genome sequencing, a high-throughput sequencing method for the whole genome of HAdV-3 based on multiplex PCR was established.Methods:Multiplex PCR primers suitable for the whole genome amplification of HAdV-3 were designed. The whole genome sequence of HAdV-3 was amplified by multiplex PCR, and the specificity of the amplification product was preliminarily verified by agarose gel electrophoresis. High-throughput sequencing of the multiplex PCR products was performed using Illumina second-generation sequencing. After obtaining the sequence, software such as CLC and IGV were used to analyze the effective data amount, average sequencing depth, and whole genome coverage obtained by high-throughput sequencing, then the sequencing quality was evaluated. Based on the whole genome sequencing result, a phylogenetic tree was constructed to confirm the virus type and analyze homology of the sequences, and then the accuracy of this method was evaluated.Results:A total of 70 (35 pairs) multiplex PCR amplification primers for the whole genome of HAdV-3 were designed, with amplicon size of approximately 1 200 bp. And the expected whole genome coverage could reach 99.8% (with a total genome length of approximately 35 240 bp). Agarose gel electrophoresis analysis showed that the size of the multiplex PCR products was consistent with expectations, and the amplification specificity was high. The high-throughput sequencing result showed that the whole genome sequences obtained by this method were complete and intact, and the genome coverage was consistent with expectations. Sequence quality analysis showed that the high-throughput sequencing method based on multiplex PCR could obtain more effective data and greater sequencing depth, resulting in more uniform whole genome coverage. Phylogenetic analysis showed that the evolutionary typing result of viral DNA sequenced after multiplex PCR amplification were consistent with those of viral DNA sequenced directly and had high homology, indicating that the multiplex PCR method had high amplification fidelity and the results obtained in combination with high-throughput sequencing were accurate.Conclusions:A high-throughput sequencing method for the whole genome of HAdV-3 based on multiplex PCR was established in this study successfully. This method could improve the efficiency and success rate of HAdV-3 whole genome sequencing, aiming to provide better technical support and reference for HAdV-3 pathogen surveillance and epidemic source-tracing based on whole genome sequencing.

Objective:To understand the genome characteristics of group A rotavirus (RVA) strains among hospitalized children under 5 years of age with acute diarrhea in Fuzhou sentinel hospital in 2020.Methods:The ELISA method was used for screening RVA-positive stool samples of hospitalized children under 5 years of age, then 11 gene segments of RVA-positive samples were sequenced and typed after amplification by RT-PCR, and their homology and phylogenetic analysis were performed by Molecular Evolutionary Genetics Analysis (MEGA).Results:Twenty RVA whole genome sequences were successfully obtained, including 4 kinds of G/P gene combinations-G9P[8] (55%), G3P[8] (25%), G8P[8] (15%) and G2P[4] (5%). DS-1-like reassortant strains accounted for 40% of the whole genomes. Strains of the same type have high sequence homology, are closely related to the virus strains that currently circulating in the world. There are mutations at multiple important antigenic sites on VP7, VP4 and NSP4 fragments. The variation of amino acid substitutions of VP7, VP4 and NSP4 fragments is complicated, and there are many amino acid substitutions in the antigenic regions.Conclusions:G3P[8] and G8P[8] DS-1-like reassortants were detected for the first time in Fuzhou, amino acid substitutions were observed in the antigenic regions of the VP7, VP4 and NSP4 gene. Due to the emergence of uncommon DS-1-like reassortant strains and multiple important antigenic regions substitutions, it is necessary to continuously monitoring genome characteristics of RVA strains to provide scientific evidence for the pandemic prevention and vaccine immunization strategies.

Objective:To investigate the whole genome characteristics and mutation of imported 2019-nCoV and trace potential source at the genomics level, the viral genomes from the specimens of a cluster of COVID-19 cases which were imported from a freighter were directly sequenced and determined by high-throughput sequencing technology and bioinformatics analysis.Methods:Throat swab specimens from the 8 confirmed cases of COVID-19 from the same freighter were collected to perform the whole genome sequencing for 2019-nCoV using the targeted genome amplification, combined with Ion S5 next-generation sequencing (NGS) technology. Varieties of online virus analysis platforms was used to classify the viruses and analyze mutation sites in the whole genome. Phylogenetic analysis software was used to construct a phylogenetic tree for the viruses, combined with the epidemiological data of the case, to speculate about the source of the viruses.Results:Eight complete genome sequences of 2019-nCoV was successfully obtained. The complete genomes of the viruses were 29, 822-29, 865 bp in length, with the average sequencing depth of 11 928×-33 588× and the coverage of 99.73%-99.87%; the result of Pangolin classification showed that all the eight 2019-nCoV genomes belong to VOC/Delta (B.1.617.2) lineage. The result of whole genome mutation analysis showed that compared with the Wuhan reference strain, the median number of nucleotide mutations in the eight 2019-nCoV genome sequences was 35 (31 to 38), and the median number of amino acid mutations was 26 (24-28); the mutation sites were distributed in 8 gene coding regions (ORF1a, ORF1b, S, ORF3a, M, ORF7a, ORF8, N). Further analysis revealed that eight 2019-nCoV genomes contained 23 characteristic mutation sites belonging to the Delta (B.1.617.2·AY.2) variants of 2019-nCoV. Since the mutation sites among the eight 2019-nCoV genomes were not completely overlapped, and the epidemiological survey report showed that the freighter stopped at multiple ports and had personnel alternation, it was speculated that the clustered COVID-19 cases might have different origins. Phylogenetic analysis showed that all the eight 2019-nCoV genomes were on the AY.2 sub-lineage of the B. 1.617.2 lineage. This result was consistent with the result of Pangolin classification and mutation analysis.Conclusions:In this study, 8 whole genome sequences of Delta variants were obtained through NGS technology from the clustered COVID-19 cases which were imported from a freighter. The sequencing method and analysis result in this study could provide reference for the 2019-nCoV mutation analysis and tracing the source of the COVID-19 cases for the prevention and control of the COVID-19 epidemic.

Objective:To investigate the prevalence and genotypes of parechovirus A (PeV-A) in hospitalized children with acute gastroenteritis under 5 years of age in Fuzhou, China.Methods:We performed a real-time RT-PCR to screen for PeV-A from stool samples and amplify VP3/VP1 junction region by nested RT-PCR to identify PeV-A type.Results:The result showed that 28 of 316 (8.86%) children with acute gastroenteritis were positive for PeV-A, two cases were co-infected with calicivirus, none with rotavirus, adenovirus or astrovirus. Totally 21 cases were PeV-A1, 1 case was PeV-A3 and 2 cases were PeV-A6 among 24 cases who were positive for PeV-A; PeV-A1 was the most prevalent strain with the proportion of 87.5%. The infection with PeV-A mainly occurred in children under 1 year of age and was prevalent between August and October.Conclusions:PeV-A1 was a main pathogen in PeV-A in hospitalized children with acute gastroenteritis in Fuzhou, and the children under 1 year of age were the main target population of PeV-A1.

Objective To identify mutations in the NEMO gene in a family with incontinentia pigmenti.Methods Clinical data were collected from the proband,and peripheral blood samples were obtained from the proband,her parents and 200 healthy controls.Multiplex PCR was performed to detect heterozygous deletion of exons 4-10 of the NEMO gene in the blood samples of the proband and her parents.Then,PCR was performed to amplify exons 2,3-10 of the NEMO gene in all the blood samples,and all exons in the gene coding region and their flanking sequences were subjected to DNA sequencing.DNA was extracted from paraffin-embedded lesional tissue of the proband's father,and PCR was performed to amplify exon 10 of the NEMO gene and its flanking sequence followed by DNA sequencing.Results The deletion of exons 4-10 of the NEMO gene was undetected in the peripheral blood of the proband or her father.Sanger sequencing showed that there was a heterozygous mutation c.1236dupA in exon 10 of the NEMO gene in the peripheral blood of the proband,which led to a mutation in amino acid residues (p.H413fs*7).The c.1236dupA mutation was not found in the peripheral blood of the proband's parents,while a mosaic mutation c.1236dupA was detected in the DNA from lesional tissues of the proband's father.Conclusion The mutation c.1236dupA in the NEMO gene may be the underlying cause of incontinentia pigmenti in the proband and her father.

Objective To investigate clinical features and detect mutations in a case of tuberous sclerosis complex (TSC) caused by a somatic mosaic mutation in the TSC2 gene.Methods Peripheral blood samples were obtained from a patient with suspected TSC,his parents,and 200 unrelated healthy controls.Genomic DNA was extracted from these blood samples,polymerase chain reaction (PCR)and nextgeneration sequencing were performed to amplify all the exons and their flanking sequences of the TSC 1 and TSC2 genes followed by DNA sequencing,so as to identify mutations in the TSC 1 and TSC2 genes.DNA was also extracted from lesional skin tissues of the patient,and PCR was conducted to amplify the target fragment of the TSC2 gene followed by DNA sequencing.Results The patient clinically presented with facial angiofibroma,depigmented patches on the waist,periungual fibroma and angioleio-myolipoma of the kidney,which were consistent with the diagnosis of TSC.A mutation c.5130_5131insT(p.V1711Cfs* 18) was identified in the TSC2 gene in the patient.A higher frequency of the mutation was found in the DNA of the tumor tissue than in that of the peripheral blood.No such a mutation was found in his parents'DNA,unrelated healthy controls or any public database.Conclusion The somatic mosaic mutation c.5130_513 1insT in the TSC2 gene is responsible for the phenotype of TSC in the patient.

Objective To detect mutations of the ABCA12 gene in 2 Chinese families with autosomal recessive congenital ichthyosis (ARCI).Methods According to the typical clinical manifestations,two probands were diagnosed with ARCI.DNA was extracted from the peripheral blood samples collected from the patients and their parents.High-throughput sequencing was conducted by using multi-gene array for genetic skin disorders to determine mutation sites in the probands,and then DNA isolated from the probands and their parents were bidirectionally verified by Sanger sequencing.Results Two compound heterozygous mutations (c.2759A＞G and c.7004A＞G) in the ABCA12 gene were found in the proband 1,and another two compound heterozygous mutations (c.6163_6164insT and c.7406G＞A) were identified in the proband 2.The parents of the two probands were heterozygous carriers of one of the two mutations in the ABCA12 gene.Function prediction for the 4 mutations showed that all of the 3 missense mutations (c.2759A＞G,c.7004A＞G and c.7406G＞A) may exert pathogenic effect,and fragnin encoded by the frameshift mutation c.6163_6164insT may also affect protein function,c.2759A＞G and c.6163_6164insT were newly identified mutation sites.Conclusion The compound heterozygous mutations in the ABCA 12 gene are the causative mutations responsible for ARCI in the two probands of the two pedigrees.