ObjectiveAngelicae Sinensis Radix， a commonly used medicinal herb with both medicinal and edible properties， is frequently adulterated in the market， severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature， quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant， "Tu Danggui" （Angelica gigas）， was conducted to establish a herbal quantitative molecular detection （Herb-Q） method for Angelicae Sinensis Radix and its processed products， providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism （SNP） loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations （linearity， limit of quantification， limit of detection， and reproducibility） by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios （15%， 25%， and 35%） was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix， based on the principles of intraspecies conservation， interspecies specificity， and meeting the requirements of pyrophosphate high-throughput sequencing， it was determined that 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 and 38 592^nd locus （T/C） in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai， respectively. The linear relationship R² of the Herb-Q method established by selecting the specific 9 674^th locus （A/G） of Angelicae Sinensis Radix was 0.997 4 （R²>0.99）， indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%， exhibiting excellent reproducibility ［relative standard deviation（RSD）<2.0%］. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder， with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability， objectivity， and accuracy for Angelicae Sinensis Radix and A. gigas， and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives， including traditional Chinese medicinal preparations， dietary supplements， and nutraceuticals.

ObjectiveAngelicae Sinensis Radix， a commonly used medicinal herb with both medicinal and edible properties， is frequently adulterated in the market， severely affecting the clinical efficacy of preparations. While qualitative identification techniques for adulterants and counterfeits are now relatively mature， quantitative detection methods for adulterated processed products remain unexplored. Quantitative detection research of Angelicae Sinensis Radix and its primary closely related adulterant， "Tu Danggui" （Angelica gigas）， was conducted to establish a herbal quantitative molecular detection （Herb-Q） method for Angelicae Sinensis Radix and its processed products， providing a model for the establishment of quantitative detection technologies for Angelicae Sinensis Radix and related health products. MethodsThe specific single-nucleotide polymorphism （SNP） loci of Angelicae Sinensis Radix and Angelica gigas Nakai were screened based on the complete chloroplast genome sequence. The specific SNP loci of Angelicae Sinensis Radix were selected for quantitative methodological investigations （linearity， limit of quantification， limit of detection， and reproducibility） by mixing the powder of the herbs with different adulteration ratios. Huoxue Zhitong powder with three distinct adulteration ratios （15%， 25%， and 35%） was utilized to ascertain the precision of the Herb-Q method for the quantitative detection of Chinese patent medicines containing Angelicae Sinensis Radix. ResultsBy comparing the 123 chloroplast genome sequences of Angelicae Sinensis Radix， based on the principles of intraspecies conservation， interspecies specificity， and meeting the requirements of pyrophosphate high-throughput sequencing， it was determined that 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 and 38 592^nd locus （T/C） in the chloroplast genome sequence NC_029393.1 could be the exclusive molecular identification loci of Angelicae Sinensis Radix and Angelica gigas Nakai， respectively. The linear relationship R² of the Herb-Q method established by selecting the specific 9 674^th locus （A/G） of Angelicae Sinensis Radix was 0.997 4 （R²>0.99）， indicating an excellent linear relationship. The limits of quantification and detection were established at 2.0%， exhibiting excellent reproducibility ［relative standard deviation（RSD）<2.0%］. The established quantitative system based on the Herb-Q method detected the adulteration amount of counterfeit A. gigas in the Huoxue Zhitong powder， with an average deviation of 1.3% for three molecular quantitative replicates. ConclusionThis research demonstrates that the Herb-Q quantitative detection method established based on the 9 674^th locus （A/G） in the chloroplast genome sequence NC_042826.1 of Angelicae Sinensis Radix has good applicability， objectivity， and accuracy for Angelicae Sinensis Radix and A. gigas， and its processed products. This method has the capacity to provide technical support for the quantitative detection of commercially available Angelicae Sinensis Radix derivatives， including traditional Chinese medicinal preparations， dietary supplements， and nutraceuticals.

Objective:To investigate the curative effects of the ulnar artery perforator flap carrying 2 and more perforators of same source artery on reconstruction of defects in hand.Methods:A retrospective observational study was conducted on 13 patients with hand injuries combined with bone or tendon exposure and met the inclusion criteria of the study were admitted to the Department of Hand Surgery, Wuxi NO.9 People’s Hospital Affiliated to Soochow University between April 2022 and September 2023. The patients were 8 males and 5 females with a mean age of 45.7 (25-67) years. Seven hand injuries were caused by mechanical crushing, 2 by hot crushing and 4 by machine strangulation. After debridement, the sizes of defect were found at 5.4 cm×5.1 cm to 8.2 cm×7.5 cm. Dopplor ultrasound was applied to locate the perforators before surgery. Perforator flaps carrying cutaneous branches of ulnar artery proximal to the wrist were designed for reconstruction of the defects in hand. The flaps were 6.0 cm×5.5 cm-8.5 cm×8.0 cm in size. Donor sites were covered by the ulnar artery perforator flaps sized 5.3 cm×2.7 cm-8.2 cm×4.0 cm and carryied 2 and more perforators of the same source artery. During harvest of the flaps, the number and calibre of perforators carried by flap, the calibre of the main trunk of the superior carpal branch of the ulnar artery and the length of its pedicle were recorded. A total of 35 perforators with 0.35-0.95 mm in calibre carried by 13 perforator flaps of ulnar artery were successfully retained. Five of the 13 flaps carried the perforators with a calibre smaller than 0.50 mm. Overall, there were 7 flaps with 2 perforators per flap, 4 with 3 perforators per flap, 1 with 4 perforators and 1 with 5 perforators. The cutaneous branch of ulnar artery in a caliber of 0.75-1.35 mm and proximal to the wrist was dissected as the vascular pedicle with 30.0-45.0 mm in length. All patients were included in the schedueled postoperative at outpatient clinic to observe the appearance and sensation of the flaps, and the occurrence of complication.Results:All flaps survived and the wounds healed at first intention without vascular compromises, wound dehiscence or obvious swelling. All patients received 8 - 20 months of follow-up, with 15 months in average. Flaps presented good appearance and colour, with soft texture and without bloating. TPD of the first flap was 8-18 mm, with an average of 10.5 mm±0.5 mm and that of the second or relay flap was 7-15 mm, with an average of 9.5 mm±0.5 mm. According to British Medical Research Council (BMRC) system, the sensory evaluation of the recipient sites achieved S 4 in 5 flaps, S 3 in 9 flaps and S 2 in 12 flaps. Conclusion:The ulnar artery perforator flap with 2 and more perforators of the same source artery has sufficient and reliable blood supply and is effective in reconstruction of hand defects.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

Objective:To investigate the curative effects of the ulnar artery perforator flap carrying 2 and more perforators of same source artery on reconstruction of defects in hand.Methods:A retrospective observational study was conducted on 13 patients with hand injuries combined with bone or tendon exposure and met the inclusion criteria of the study were admitted to the Department of Hand Surgery, Wuxi NO.9 People’s Hospital Affiliated to Soochow University between April 2022 and September 2023. The patients were 8 males and 5 females with a mean age of 45.7 (25-67) years. Seven hand injuries were caused by mechanical crushing, 2 by hot crushing and 4 by machine strangulation. After debridement, the sizes of defect were found at 5.4 cm×5.1 cm to 8.2 cm×7.5 cm. Dopplor ultrasound was applied to locate the perforators before surgery. Perforator flaps carrying cutaneous branches of ulnar artery proximal to the wrist were designed for reconstruction of the defects in hand. The flaps were 6.0 cm×5.5 cm-8.5 cm×8.0 cm in size. Donor sites were covered by the ulnar artery perforator flaps sized 5.3 cm×2.7 cm-8.2 cm×4.0 cm and carryied 2 and more perforators of the same source artery. During harvest of the flaps, the number and calibre of perforators carried by flap, the calibre of the main trunk of the superior carpal branch of the ulnar artery and the length of its pedicle were recorded. A total of 35 perforators with 0.35-0.95 mm in calibre carried by 13 perforator flaps of ulnar artery were successfully retained. Five of the 13 flaps carried the perforators with a calibre smaller than 0.50 mm. Overall, there were 7 flaps with 2 perforators per flap, 4 with 3 perforators per flap, 1 with 4 perforators and 1 with 5 perforators. The cutaneous branch of ulnar artery in a caliber of 0.75-1.35 mm and proximal to the wrist was dissected as the vascular pedicle with 30.0-45.0 mm in length. All patients were included in the schedueled postoperative at outpatient clinic to observe the appearance and sensation of the flaps, and the occurrence of complication.Results:All flaps survived and the wounds healed at first intention without vascular compromises, wound dehiscence or obvious swelling. All patients received 8 - 20 months of follow-up, with 15 months in average. Flaps presented good appearance and colour, with soft texture and without bloating. TPD of the first flap was 8-18 mm, with an average of 10.5 mm±0.5 mm and that of the second or relay flap was 7-15 mm, with an average of 9.5 mm±0.5 mm. According to British Medical Research Council (BMRC) system, the sensory evaluation of the recipient sites achieved S 4 in 5 flaps, S 3 in 9 flaps and S 2 in 12 flaps. Conclusion:The ulnar artery perforator flap with 2 and more perforators of the same source artery has sufficient and reliable blood supply and is effective in reconstruction of hand defects.

BACKGROUND:At present,the treatment methods for necrotizing fasciitis mostly use negative pressure sealing suction after thorough debridement.This method requires repeated debridement to completely remove necrotic infected tissue,causing serious physical and economic burdens to patients. OBJECTIVE:To introduce a rare clinical case of calf compartment syndrome caused by diabetic foot necrotizing fasciitis,and summarize the clinical experience of using antibiotic-loaded bone cement for treatment and comprehensive management. METHODS:A total of 6 patients with calf compartment syndrome caused by diabetic necrotizing fasciitis admitted to Wuxi 9th Affiliated Hospital of Soochow University from August 2017 to August 2020 were selected,including 5 males and 1 female with an average age of 54 years.During the perioperative period,the patients'general condition was evaluated and systemic nutritional support treatment was given.In the first stage,all patients received complete debridement to control infection,antibiotic-loaded bone cement packing,and negative pressure sealed drainage.In the second stage,bone cement was removed and wound repair was performed.The wound healing,as well as the occurrence of redness,swelling,and exudation was observed during the follow-up. RESULTS AND CONCLUSION:(1)The wounds of four patients were fresh after twice antibiotic-loaded bone cement packing,and the membrane formation was good,and one patient was good after three times of antibiotic-loaded bone cement packing,and the wounds of all five patients healed well after the second stage of skin grafting.Due to the difficulty in maintaining intraoperative blood pressure and infection in all four compartments of the lower leg,a patient underwent emergency knee amputation.Meanwhile,the stump wound was placed with antibiotic-loaded bone cement.The wound was closed directly after the secondary bone cement was removed,and the wound healed in the first stage.(2)The six patients were followed up for 6-24 months after discharge.At the last follow-up,all six patients had good wound healing and no symptoms such as redness,swelling,and exudation.The quality of life of the patients was significantly improved,and all of them were satisfied with the curative effect.(3)The occurrence of calf compartment syndrome should be vigilant when diabetic foot necrotizing fasciitis is highly suspected.Early diagnosis and timely incision decompression are of great importance.Besides,the application of antibiotic-loaded bone cement in the treatment of calf compartment syndrome caused by diabetic necrotizing fasciitis has a good short-term effect.

Objective:To explore the clinical efficacy of antibiotic bone cement combined with vacuum sealing drainage (VSD) in treating diabetes mellitus complicated with necrotizing fasciitis.Methods:The retrospective observational study approach was used. From January 2020 to March 2022, 12 patients with type 2 diabetes complicated with necrotizing fasciitis who met the inclusion criteria were admitted to Wuxi Ninth People's Hospital, including 7 males and 5 females, aged 27 to 76 years. The initial diagnosis of lesions was in the lower limbs. After admission, bedside incision and drainage were performed timely, and a sample of wound exudate was collected for microbial cultivation. At the same time, the comprehensive supportive treatment was performed. At stage Ⅰ, debridement was performed, and the skin and soft tissue defect area was 40 cm×15 cm to 80 cm×25 cm after debridement. The dead space was filled with bone cement containing gentamicin and vancomycin and VSD was performed. After there was no obvious infection on the wound, the antibiotic bone cement was removed and wound repair surgery was performed at stage Ⅱ. The times of debridement, amputation, infection control, wound treatment method and wound healing at stage Ⅱ, total hospitalization day, and recurrence of necrotizing fasciitis during follow-up after the stage Ⅱ surgery. At the last follow-up, the walking function of patients was evaluated according to the scoring standards of American Orthopedic Foot and Ankle Association (AOFAS).Results:Eleven patients had wound infection control with one debridement surgery and did not undergo amputation surgery; one patient had significant foot gangrene, and the infection was controlled after one debridement and amputation of the gangrenous limb. Blood routine and infection indicators gradually returned to normal within 7 days after surgery. At stage Ⅱ, the wounds in 4 patients were sutured directly, the wounds in 6 patients were repaired with full-thickness inguinal skin graft, while the wounds in 2 patients were repaired with pedicled or tongue-shaped flaps at the wound edge. The wounds healed well after surgery, with no ulceration. The total hospitalization day of patients was 20 to 45 days. Follow-up for 3 to 24 months after stage Ⅱ surgery showed no recurrence of necrotizing fasciitis in any patient. At the last follow-up, the walking function was evaluated as excellent in 10 cases and good in 2 cases according to the AOFAS scoring standard.Conclusions:Antibiotic bone cement combined with VSD used in treating type 2 diabetes complicated with necrotizing fasciitis can effectively control infection and reduce the times of debridement, with good wound healing and walking function after surgery.