Objective:To explore the clinical efficacy of anterolateral thigh perforator flap (ALTPF) in treatment of extensive wound of Degree IV burns in limbs.Methods:A retrospective analysis was conducted on 9 patients who had extensive wound of Degree IV burns in limbs caused by stove burns admitted to Department of Burns and Plastic Surgery, the 988th Hospital of the Joint Logistics Support Force of the Chinese PLA between January 2017 and January 2024. Among the patients, there were 8 males and 1 female, aged between 36 and 63 years. Three patients had the wound from anterior leg to dorsal foot, 3 from leg down to ankle, 1 from forearm to hand and 2 from arm to forearm. Area of burns ranged from 20 cm × 15 cm to 30 cm × 25 cm, and all patients were treated by free ALTPF. According to whether the main artery at the recipient site was feasible for direct anastomosis with the vessels carried in flap, 4 patients were treated by bilateral parallel ALTPFs, and 5 were treated by unilateral ultra-long internally supercharged ALTPF. A total of 13 ALTPFs were harvested, with individual flap size at 20 cm × 8 cm to 46 cm × 12 cm. Donor sites were directly sutured. Time for flap harvesting, flap survival and wound healing time were records. Scheduled postoperative follow-up was conducted at outpatient clinic and via telephone interviews to evaluate functional recovery. Follow-up assessments included evaluation of flap condition, two-point discrimination (TPD), recovery of joint function at recipient sites, flap appearance and donor site recovery.Results:The time for flap harvest was 1.0 to 4.5 hours. All 13 ALTPFs successfully survived. The time from surgery to healing of recipient sites was 18 to 72 days, and all donor sites healed. Over the postoperative follow-up that lasted for 6 to 34 months, the recipient sites had found with good cosmetic outcomes, without osteomyelitis or deep tissue infection. Four ALTPFs in 2 patients were found swelling, which were revised at 6 months after surgery. Four ALTPFs in other 2 patients had pigment deposition at edges. One ALTPF was scalded, which healed after dressing changes but left with patchy scars. The remaining ALTPFs were soft, elastic, free from pain and well-perfused, with regained protective sensation at S 3. However, all of the ALTPFs failed to detect TPD. Six patients with lower limb injuries were evaluated using the American Orthopedic Foot and Ankle Society (AOFAS) ankle hindfoot scoring system: 2 patients were rated as excellent and 4 were rated as good. Three patients with upper limb injuries were evaluated using Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association: 1 patient was rated as excellent and 2 were rated as good. Only linear scars left at the 13 donor sites, with normal blood supply to the distal limbs, and without restrictions in range of motion of knee joints nor muscle strength of quadriceps. Conclusion:The ALTPF offers advantages such as anatomical consistency, reliable blood supply and flexible combination in treatment of extensive wound of Degree Ⅳ burns in limbs. It is an ideal surgical procedure for treatment of large soft tissue defects of extremities.

Objective:To explore the clinical efficacy of anterolateral thigh perforator flap (ALTPF) in treatment of extensive wound of Degree IV burns in limbs.Methods:A retrospective analysis was conducted on 9 patients who had extensive wound of Degree IV burns in limbs caused by stove burns admitted to Department of Burns and Plastic Surgery, the 988th Hospital of the Joint Logistics Support Force of the Chinese PLA between January 2017 and January 2024. Among the patients, there were 8 males and 1 female, aged between 36 and 63 years. Three patients had the wound from anterior leg to dorsal foot, 3 from leg down to ankle, 1 from forearm to hand and 2 from arm to forearm. Area of burns ranged from 20 cm × 15 cm to 30 cm × 25 cm, and all patients were treated by free ALTPF. According to whether the main artery at the recipient site was feasible for direct anastomosis with the vessels carried in flap, 4 patients were treated by bilateral parallel ALTPFs, and 5 were treated by unilateral ultra-long internally supercharged ALTPF. A total of 13 ALTPFs were harvested, with individual flap size at 20 cm × 8 cm to 46 cm × 12 cm. Donor sites were directly sutured. Time for flap harvesting, flap survival and wound healing time were records. Scheduled postoperative follow-up was conducted at outpatient clinic and via telephone interviews to evaluate functional recovery. Follow-up assessments included evaluation of flap condition, two-point discrimination (TPD), recovery of joint function at recipient sites, flap appearance and donor site recovery.Results:The time for flap harvest was 1.0 to 4.5 hours. All 13 ALTPFs successfully survived. The time from surgery to healing of recipient sites was 18 to 72 days, and all donor sites healed. Over the postoperative follow-up that lasted for 6 to 34 months, the recipient sites had found with good cosmetic outcomes, without osteomyelitis or deep tissue infection. Four ALTPFs in 2 patients were found swelling, which were revised at 6 months after surgery. Four ALTPFs in other 2 patients had pigment deposition at edges. One ALTPF was scalded, which healed after dressing changes but left with patchy scars. The remaining ALTPFs were soft, elastic, free from pain and well-perfused, with regained protective sensation at S 3. However, all of the ALTPFs failed to detect TPD. Six patients with lower limb injuries were evaluated using the American Orthopedic Foot and Ankle Society (AOFAS) ankle hindfoot scoring system: 2 patients were rated as excellent and 4 were rated as good. Three patients with upper limb injuries were evaluated using Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association: 1 patient was rated as excellent and 2 were rated as good. Only linear scars left at the 13 donor sites, with normal blood supply to the distal limbs, and without restrictions in range of motion of knee joints nor muscle strength of quadriceps. Conclusion:The ALTPF offers advantages such as anatomical consistency, reliable blood supply and flexible combination in treatment of extensive wound of Degree Ⅳ burns in limbs. It is an ideal surgical procedure for treatment of large soft tissue defects of extremities.

Objective:To investigate the genotype and epidemiological characteristics of human metapneumovirus (HMPV) among hospitalized cases with acute respiratory infections (ARI) in children in Changchun City, Jilin Province, China.Methods:From June 2019 to June 2023, throat swabs of ARI inpatients in Changchun Children′s Hospital were collected, and their epidemiological and clinical information were also collected. Quantitative reverse transcription-PCR was used to identify HMPV-positive cases, followed by the amplification of the G gene and genetic analysis in the HMPV-positive cases.Results:A total of 3 311 children hospitalized with ARI were included in this study. Their age ranged from 0 to 17 years old, and the M ( Q1, Q3) of age was 2 (1, 3) years. About 1 811 (54.70%) cases were males. A total of 167 HMPV-positive cases were detected with a positive rate of 5.04%, of which 92.81% (155/167) were children under 5 years old. The positive rate of HMPV in 2019 was 6.37% (30/471), which dropped to the lowest in 2020 (2.31%, 10/432). The HMPV-positive rate was then rebounded in 2021 (4.70%, 60/1 277) and 2022 (4.56%, 21/461), which increased to 6.87% (46/670) in 2023. The difference in HMPV-positive rate among each year was statistically significant ( P<0.05). The prevalence peak of HMPV varied in different years, showing either a unimodal or bimodal distribution in one year. A total of 79 HMPV G gene sequences were obtained, of which subtype A and subtype B accounted for 48.10% and 51.90%, respectively. All of the subtype A sequences were clarified as A2c duplicated variants, and subtype B was mainly B2 genotype. Besides, subtypes A and B were prevalent alone or co-circulated in different years, and there was a subtype replacement pattern in HMPV. Conclusion:The positive rate of HMPV in hospitalized ARI cases in children is significantly different from 2019 to 2023 in Changchun City. Notably, there are certain switch patterns of HMPV subtypes A and B in different years.

Objective:To investigate the genotype and epidemiological characteristics of human metapneumovirus (HMPV) among hospitalized cases with acute respiratory infections (ARI) in children in Changchun City, Jilin Province, China.Methods:From June 2019 to June 2023, throat swabs of ARI inpatients in Changchun Children′s Hospital were collected, and their epidemiological and clinical information were also collected. Quantitative reverse transcription-PCR was used to identify HMPV-positive cases, followed by the amplification of the G gene and genetic analysis in the HMPV-positive cases.Results:A total of 3 311 children hospitalized with ARI were included in this study. Their age ranged from 0 to 17 years old, and the M ( Q1, Q3) of age was 2 (1, 3) years. About 1 811 (54.70%) cases were males. A total of 167 HMPV-positive cases were detected with a positive rate of 5.04%, of which 92.81% (155/167) were children under 5 years old. The positive rate of HMPV in 2019 was 6.37% (30/471), which dropped to the lowest in 2020 (2.31%, 10/432). The HMPV-positive rate was then rebounded in 2021 (4.70%, 60/1 277) and 2022 (4.56%, 21/461), which increased to 6.87% (46/670) in 2023. The difference in HMPV-positive rate among each year was statistically significant ( P<0.05). The prevalence peak of HMPV varied in different years, showing either a unimodal or bimodal distribution in one year. A total of 79 HMPV G gene sequences were obtained, of which subtype A and subtype B accounted for 48.10% and 51.90%, respectively. All of the subtype A sequences were clarified as A2c duplicated variants, and subtype B was mainly B2 genotype. Besides, subtypes A and B were prevalent alone or co-circulated in different years, and there was a subtype replacement pattern in HMPV. Conclusion:The positive rate of HMPV in hospitalized ARI cases in children is significantly different from 2019 to 2023 in Changchun City. Notably, there are certain switch patterns of HMPV subtypes A and B in different years.

In order to achieve rapid detection of duck hepatitis A virus type 3(DHAV-3),this ex-periment initially performed prokaryotic expression of the non-structural protein 3D of DHAV-3,followed by immunization of BALB/c mice with the purified protein.After immunization,mouse spleen cells were fused with myeloma cells(SP2/0)to prepare monoclonal antibodies.Subsequent-ly,a double-antibody sandwich ELISA(DAS-ELISA)detection method was established using the monoclonal antibodies,and its sensitivity,specificity,and repeatability were evaluated.Finally,the established method was applied to the detection of clinical samples and validated for compliance with the RT-PCR method.The results showed that the DHAV-3 3D protein was efficiently ex-pressed in BL21(DE3),and its specificity was confirmed by Western blot after purification.After cell fusion and three rounds of subcloning,six hybridoma cells were successfully screened and named 1A3,1B6,1C7,1D9,2A1,and 3A9.The subtype identification of the antibodies showed that 1A3 belonged to IgG2b,1B6 belonged to IgG2a,3A9 belonged to IgG3,and 1C7,1D9,and 2A1 be-longed to IgG1.After screening,the high-affinity monoclonal antibodies 1B6 and 1 A3 were selected as the capture antibody and detection antibody,respectively,and use to establish the DAS-ELISA detection method.After optimizing the reaction conditions,the optimal coating concentration of the capture antibody 1B6 was determined to be 1×10-3 g/L,and the optimal dilution of the detection antibody 1A3 was 1∶1 000.The cut-off value was established as 0.256.The sensitivity test showed that the method had a minimum detection limit of 4.0 ×10-4 g/L for the 3D protein.The repeat-ability test showed that the within-batch and between-batch coefficients of variation were both less than 9％,indicating good repeatability.The specificity test showed that the method did not show specific reactions with duck adenovirus(DAdV),muscovy duck parvovirus(MDPV),duck circo-virus(DuCV),duck plague virus(DPV),duck reovirus(DRV),or Riemerella anatipestifer(RA),but cross-reacted with Duck hepatitis a virus type 1(DHAV-1),allowing simultaneous de-tection of DHAV-3 and DHAV-1 pathogens.The DAS-ELISA method established in this experi-ment was compared with the RT-PCR method for the detection of 186 clinical samples,and the DAS-ELISA method could simultaneously identify DHAV-1 and DHAV-3,with a compliance rate of 98.9％compared to the RT-PCR method.In conclusion,the established DAS-ELISA method showed good repeatability and high sensitivity,and can be used for the diagnosis of DHAV-1 and DHAV-3,providing technical support for the epidemiological investigation and prevention of Duck Hepatitis A.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

In order to understand the prevalence of Akabane disease(AKAD)in Guizhou Province and the molecular characteristics of the isolates,the whole-genome sequence of a strain of Akabane virus(AKAV)from a bovine AKAD-positive sample was determined and analyzed.The genotype and genetic variation of the strain were also explored.Based on the conserved S sequence,a fluores-cence quantitative PCR(qPCR)detection method was established and applied for the investigation of AKAV infection status in four large-scale beef cattle farms of Guizhou.Results showed that the S,M and L fragments of the bovine strain were highly homologous to the Tianjin strain(TJ2016/China/2016)and the Australian strain(JaLAB39/Australia/1959),where they were in the same evolutionary branch and belonged to genotype Ⅱ.Sensitivity assay found that the lowest detection limit was 2.5 X 101 copies/μL.Specificity assay showed the established method detected only AKAV with no amplification on bovine bluetongue virus(BLUV),Pasteurella multocida(PM),bovine infectious rhinotracheitis virus(IBRV)and bovine Mycoplasma bovis.The variation coefficients of inter-and intra batches in the repeatability test were both lower than 2.26％.These findings illus-trated that the established qPCR method had high sensitivity,good specificity and repeatability.A total of 298 serum samples from 4 large-scale beef cattle farms in Qianxi City and Huangping County of Guizhou Province were collected and tested for AKAV by the method.Out of 298 sam-ples,25 positive samples(25/298)were detected as positive with a positive rate of 8.39％.In sum-mary,this work provided the reference data for a deep understanding of the molecular prevalence of AKAV in Guizhou Province and laid foundation for the prevention and control of AKAD.

In order to achieve rapid detection of duck hepatitis A virus type 3(DHAV-3),this ex-periment initially performed prokaryotic expression of the non-structural protein 3D of DHAV-3,followed by immunization of BALB/c mice with the purified protein.After immunization,mouse spleen cells were fused with myeloma cells(SP2/0)to prepare monoclonal antibodies.Subsequent-ly,a double-antibody sandwich ELISA(DAS-ELISA)detection method was established using the monoclonal antibodies,and its sensitivity,specificity,and repeatability were evaluated.Finally,the established method was applied to the detection of clinical samples and validated for compliance with the RT-PCR method.The results showed that the DHAV-3 3D protein was efficiently ex-pressed in BL21(DE3),and its specificity was confirmed by Western blot after purification.After cell fusion and three rounds of subcloning,six hybridoma cells were successfully screened and named 1A3,1B6,1C7,1D9,2A1,and 3A9.The subtype identification of the antibodies showed that 1A3 belonged to IgG2b,1B6 belonged to IgG2a,3A9 belonged to IgG3,and 1C7,1D9,and 2A1 be-longed to IgG1.After screening,the high-affinity monoclonal antibodies 1B6 and 1 A3 were selected as the capture antibody and detection antibody,respectively,and use to establish the DAS-ELISA detection method.After optimizing the reaction conditions,the optimal coating concentration of the capture antibody 1B6 was determined to be 1×10-3 g/L,and the optimal dilution of the detection antibody 1A3 was 1∶1 000.The cut-off value was established as 0.256.The sensitivity test showed that the method had a minimum detection limit of 4.0 ×10-4 g/L for the 3D protein.The repeat-ability test showed that the within-batch and between-batch coefficients of variation were both less than 9％,indicating good repeatability.The specificity test showed that the method did not show specific reactions with duck adenovirus(DAdV),muscovy duck parvovirus(MDPV),duck circo-virus(DuCV),duck plague virus(DPV),duck reovirus(DRV),or Riemerella anatipestifer(RA),but cross-reacted with Duck hepatitis a virus type 1(DHAV-1),allowing simultaneous de-tection of DHAV-3 and DHAV-1 pathogens.The DAS-ELISA method established in this experi-ment was compared with the RT-PCR method for the detection of 186 clinical samples,and the DAS-ELISA method could simultaneously identify DHAV-1 and DHAV-3,with a compliance rate of 98.9％compared to the RT-PCR method.In conclusion,the established DAS-ELISA method showed good repeatability and high sensitivity,and can be used for the diagnosis of DHAV-1 and DHAV-3,providing technical support for the epidemiological investigation and prevention of Duck Hepatitis A.

Lateral drainage is an important way for the middle and lower rectum. There are still disputes between Eastern and Western scholars regarding the treatment strategy of lateral lymph node metastasis. It is difficult to diagnose lateral lymph node metastasis by pathologic examination before surgery. MRI is the main method for diagnosis of lateral lymph node metastasis in rectal cancer. The diagnostic criteria for patients before and after neoadjuvant treatment are slightly different. CT, PET/CT and radiomics also have certain advantages in the diagnosis of lateral lymph node metastasis. If necessary, the combination of multiple methods can help in the diagnosis of lateral lymph node metastasis.

Objective:To investigate the clinical characteristics and prognosis of duodenal neuroendocrine neoplasms.Methods:The clinical data of 35 patients with duodenal neuroendocrine neoplasms admitted to Union Hospital, Tongji Medical College, Huazhong University of Science & Technology from Jan 2012 to Dec 2021 were retrospectively analyzed. The differences of clinical characteristics between periampullary and non-periampullary duodenal neuroendocrine neoplasms were analyzed. Kaplan-Meier curve was used for survival analysis, and the clinical factors affecting the prognosis were analyzed.Results:Of the 35 patients, 30 underwent tumor resection, 7 (23%) developed different degree of complications after operation and were improved and discharged after intervention. A total of 5 patients died during the follow-up period. Only 1 of 30 patients who underwent tumor resection died 30 months after operation due to disease progression, and the others had no recurrence or metastasis. Univariate analysis showed that tumor size, tumor grade, and tumor location were associated with the prognosis of patients (all P<0.05), and multivariate analysis showed that patients with tumors located.Away from the ampulla had a significantly better prognosis than those located around the duodenal ampulla ( P<0.01). Conclusions:Patients with duodenal neuroendocrine neoplasms have a good prognosis after complete resection; patients with duodenal neuroendocrine neoplasms located around the ampulla of Vater have a relatively poor prognosis compared with those away from the area of ampulla.