Two thousand five hundred and sixty swab samples were collected from December 2022 to December 2023 in China.PCR was used to detect FBoV and amplify its VP2 and NS1 gene cod-ing equences,and bioinformatics was used to analyze the genetic diversity of FBoV.The results showed that the total positive rate of FBoV was 4.6％(119/2 560).Genetic variation analysis showed that FBoV existed in a variety of genotypes,and FBOV-1 was the main epidemic type in China.The 15 FBoV-1 strains,four FBoV-2 strains and one FBoV-3 strains identified in this study were genetically close to the strains identified in China,the United States,Thailand,Australia and Portugal.Sequence analysis showed that the identities of amino acid sequence of NS1 and VP2 genes between the sequenced strains and the reference strains were 59.13％-99.25％and 96.41％-100.00％,respectively.The amino acid identities of NS1 and VP2 among the newly sequenced FBoV strains were 60.00％-100.00％and 96.41％-100.00％,respectively,which indicated that the FBov strains circulating in China had great genetic diversity.This study enriched the data for elucidating the epidemic status of FBoV in China,and provided the basis for the subsequent diag-nosis,prevention and control of FBoV.

Objective:To study the clinical outcome of palmar approach to on-top plasty in the treatment of unequal radial polydactyly.Methods:A retrospective analysis was conducted on patients with unequal radial polydactyly in which neither thumb duplicates possess both well-developed proximal and distal components. All the patients underwent palmar approach to on-top plasty between September 2013 and September 2023 at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine and Shanxi Children’s Hospital. At the final follow-up, the active flexion-extension range of motion of the first metacarpophalangeal and interphalangeal joints was measured, along with grip and pinch strength. The Vancouver scar scale was used to assess scar formation (total score 0-15, with higher scores indicating more severe scarring). Additionally, the pediatric quality of life inventory (total score 0-100, with higher scores indicating better quality of life, evaluated by the children themselves or by their parents/guardians on their behalf) and the satisfaction questionnaire (total score ranges from 1 to 5, with higher scores indicating greater satisfaction) were used to assess the patients’ daily quality of life and parents’satisfaction.Results:A total of 28 pediatric patients were included, comprising 15 males and 13 females, with an average age of 15 months (range: 10-36 months). All cases presented unilateral radial polydactyly, including 18 right hands and 10 left hands. The mean follow-up duration was 4.2 years (range: 1.5-6.0 years). All incisions healed in one stage, and the skin flaps survived well without infection. Joint stability was maintained without any deviation. All reconstructed thumbs demonstrated satisfactory aesthetic appearance. The reconstructed thumbs demonstrated mean active flexion-extension arcs of 76° (range 61°-85°) at the first metacarpophalangeal joint, and 42° (range 21°-65°) at the interphalangeal joint. The grip strength of the hand was 82% (63%-90%) of the normal opposite hand. The key, tip and tripod pinch strength were 76%(66%-83%), 77%(66%-85%) and 79%(68%-87%) of the normal opposite hand, respectively. No scar hyperplasia was found in all cases, and the Vancouver scar scale score was 1.2(0-3) points. The self-assessment score of the children on the pediatric quality of life inventory was 86 points, and the score of the parents/guardians was 91 points. In terms of family satisfaction, 23 patients scored 5, and 5 patients scored 4.Conclusion:The use of palmar approach to on-top plasty to treat unequal radial polydactyly can achieve functional and aesthetic results.

Two thousand five hundred and sixty swab samples were collected from December 2022 to December 2023 in China.PCR was used to detect FBoV and amplify its VP2 and NS1 gene cod-ing equences,and bioinformatics was used to analyze the genetic diversity of FBoV.The results showed that the total positive rate of FBoV was 4.6％(119/2 560).Genetic variation analysis showed that FBoV existed in a variety of genotypes,and FBOV-1 was the main epidemic type in China.The 15 FBoV-1 strains,four FBoV-2 strains and one FBoV-3 strains identified in this study were genetically close to the strains identified in China,the United States,Thailand,Australia and Portugal.Sequence analysis showed that the identities of amino acid sequence of NS1 and VP2 genes between the sequenced strains and the reference strains were 59.13％-99.25％and 96.41％-100.00％,respectively.The amino acid identities of NS1 and VP2 among the newly sequenced FBoV strains were 60.00％-100.00％and 96.41％-100.00％,respectively,which indicated that the FBov strains circulating in China had great genetic diversity.This study enriched the data for elucidating the epidemic status of FBoV in China,and provided the basis for the subsequent diag-nosis,prevention and control of FBoV.

Objective:To study the clinical outcome of palmar approach to on-top plasty in the treatment of unequal radial polydactyly.Methods:A retrospective analysis was conducted on patients with unequal radial polydactyly in which neither thumb duplicates possess both well-developed proximal and distal components. All the patients underwent palmar approach to on-top plasty between September 2013 and September 2023 at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine and Shanxi Children’s Hospital. At the final follow-up, the active flexion-extension range of motion of the first metacarpophalangeal and interphalangeal joints was measured, along with grip and pinch strength. The Vancouver scar scale was used to assess scar formation (total score 0-15, with higher scores indicating more severe scarring). Additionally, the pediatric quality of life inventory (total score 0-100, with higher scores indicating better quality of life, evaluated by the children themselves or by their parents/guardians on their behalf) and the satisfaction questionnaire (total score ranges from 1 to 5, with higher scores indicating greater satisfaction) were used to assess the patients’ daily quality of life and parents’satisfaction.Results:A total of 28 pediatric patients were included, comprising 15 males and 13 females, with an average age of 15 months (range: 10-36 months). All cases presented unilateral radial polydactyly, including 18 right hands and 10 left hands. The mean follow-up duration was 4.2 years (range: 1.5-6.0 years). All incisions healed in one stage, and the skin flaps survived well without infection. Joint stability was maintained without any deviation. All reconstructed thumbs demonstrated satisfactory aesthetic appearance. The reconstructed thumbs demonstrated mean active flexion-extension arcs of 76° (range 61°-85°) at the first metacarpophalangeal joint, and 42° (range 21°-65°) at the interphalangeal joint. The grip strength of the hand was 82% (63%-90%) of the normal opposite hand. The key, tip and tripod pinch strength were 76%(66%-83%), 77%(66%-85%) and 79%(68%-87%) of the normal opposite hand, respectively. No scar hyperplasia was found in all cases, and the Vancouver scar scale score was 1.2(0-3) points. The self-assessment score of the children on the pediatric quality of life inventory was 86 points, and the score of the parents/guardians was 91 points. In terms of family satisfaction, 23 patients scored 5, and 5 patients scored 4.Conclusion:The use of palmar approach to on-top plasty to treat unequal radial polydactyly can achieve functional and aesthetic results.

In order to prepare monoclonal antibodies with blocking activity against feline TNF-α,this study successfully constructed,expressed and purified the recombinant plasmid pET-28a-sTNFα based on the soluble feline TNF-α(sTNFα)gene,and further investigated the induced ex-pression.The conditions were explored and optimized to identify its biological activity;secondly,the feline TNF-α recombinant protein was used as an immunogen for mouse immunization,after cell fusion,screening of blocking active hybridoma cells and ascites preparation,the obtained mon-oclonal antibodies were tested.The results showed that the pET-28a-sTNFα plasmid was success-fully constructed and the bioactive feline TNF-α recombinant protein was expressed in E.coli sys-tem.The molecular weight was 34 kDa and the 50％inhibitory concentration was 1.22 pg/L.Three monoclonal antibodies(A6-B7-9,H5-E2-94 and C8-A10-100)with blocking activity were success-fully screened out.The results of Western blot showed that all the three mAbs could specifically bind to TNF-α with a titer of 1:512 000.When the concentration of the three mAbs was 100 mg/L,the inhibitory effect on TNF-α was the strongest.In this study,we screened antibodies that can block the activity of cat TNF-α,in order to provide novel,safe and effective candidate drugs for the treatment of TNF-α mediated diseases in cats.

BACKGROUND: Corneal disease is second only to cataract considered as the leading cause of blindness in the world, with high morbidity. Construction of corneal substitutes In Vitro by tissue engineering technology to achieve corneal regeneration has become a research hotspot in recent years. We conducted in-depth research on the biocompatibility, physicochemical and mechanical properties of rat bone marrow mesenchymal stem cells (rBM-MSCs)-seeded gelatin methacrylate (GelMA) as a bioengineered cornea. METHODS: Four kinds of GelMA with different concentrations (7, 10, 15 and 30%) were prepared, and their physicchemical, optical properties, and biocompatibility with rBM-MSCs were characterized. MTT, live/dead staining, cell morphology, immunofluorescence staining and gene expression of keratocyte markers were performed. RESULTS: 7%GelMA hydrogel had higher equilibrium water content and porosity, better optical properties and hydrophilicity. In addition, it is more beneficial to the growth and proliferation of rBM-MSCs. However, the 30%GelMA hydrogel had the best mechanical properties, and could be more conducive to promote the differentiation of rBM-MSCs into keratocyte-like cells. CONCLUSION As a natural biological scaffold, GelMA hydrogel has good biocompatibility. And it has the ability to promote the differentiation of rBM-MSCs into keratocyte-like cells, which laid a theoretical and experimental foundation for further tissue-engineered corneal stromal transplantation, and provided a new idea for the source of seeded cells in corneal tissue engineering.

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

A 27﹣year﹣old male patient underwent left maxillary masses excision for periapical cysts in the left maxillary under general anesthesia. During the surgery,a venous access was established from the peripheral vein of the patient′s left lower extremity,and anesthetic drugs(propofol injection,succinylcholine chloride injection,remifentanil hydrochloride for injection,dexmedetomidine hydrochloride injection)were given. On day 2 after surgery,the patient developed topical redness and swelling along the blood vessel on his left lower extremity,which gradually developed into a cord﹣like redness and swelling,with increased blood vessel stiffness. He was diagnosed with thrombophlebitis of the left great saphenous vein after a ultrasound examination of the blood vessel. Treatments such as subcutaneous injection of enoxaparin sodium, oral administration of extract of horse chestnut seeds, topical mucopolysaccharide polysulfate cream, physiotherapy,and etc. were given. On day 10 after surgery,the redness and swelling on the patient′s left lower extremity disappeared,and the blood vessel was softened. At 2 months of follow﹣up,ultrasound examination revealed partial recanalization of the left venous trunk( the calf part ) of saphenous vein. According to the drug labels and literature reports,it was considered that the patient′s thrombophlebitis might be associated with propofol injection.

A 27﹣year﹣old male patient underwent left maxillary masses excision for periapical cysts in the left maxillary under general anesthesia. During the surgery,a venous access was established from the peripheral vein of the patient′s left lower extremity,and anesthetic drugs(propofol injection,succinylcholine chloride injection,remifentanil hydrochloride for injection,dexmedetomidine hydrochloride injection)were given. On day 2 after surgery,the patient developed topical redness and swelling along the blood vessel on his left lower extremity,which gradually developed into a cord﹣like redness and swelling,with increased blood vessel stiffness. He was diagnosed with thrombophlebitis of the left great saphenous vein after a ultrasound examination of the blood vessel. Treatments such as subcutaneous injection of enoxaparin sodium, oral administration of extract of horse chestnut seeds, topical mucopolysaccharide polysulfate cream, physiotherapy,and etc. were given. On day 10 after surgery,the redness and swelling on the patient′s left lower extremity disappeared,and the blood vessel was softened. At 2 months of follow﹣up,ultrasound examination revealed partial recanalization of the left venous trunk( the calf part ) of saphenous vein. According to the drug labels and literature reports,it was considered that the patient′s thrombophlebitis might be associated with propofol injection.