This study was performed to screen the sequence of specific nanobody targeting SARS-CoV-2 S protein,and to express and purify recombinant anti-SARS-CoV-2 nanobodies,and evaluate their characteristics and application potential.Based on the previously constructed natural bacteriophage nanobody display library,the anti-SARS-CoV-2 S protein nanobody sequences were screened,with biotinylated SARS-CoV-2 S protein receptor-binding domain antigen as target,by biotin-streptavidin liquid phase screening method,phage-ELISA and sequencing.Recombinant anti-SARS-CoV-2 S protein nanobody expression vectors were constructed,the protein was induced by IPTG and then purified,and their characteristics and application potential were analyzed by SDS-PAGE,Westem blot and ELISA methods.Two anti-SARS-CoV-2 S protein nanobody sequences were successfully obtained,and the corresponding prokaryotic expression bacteria were constructed.The recombinant anti-SARS-CoV-2 S protein nanobody was expressed and purified successfully,which can specifically bind to SARS-CoV-2 S protein.In conclusion,the recombinant anti-SARS-CoV-2 S protein nanobodies have screened and expressed successfully,which provides a basis for its application in SARS-CoV-2 detection and treatment,and facilitates related researches.

Objective:To assess the efficacy and safety of common medication treatments on Kaposiform hemangioendothelioma (KHE) and tufted angioma (TA).Methods:PubMed, Embase, Web of Science, CNKI and Wanfang database were searched to find out the observational studies on medication treatment of KHE and TA. R-3.6.2 was used for calculate the pooled response rate and pooled adverse events rate. Meta analyses were performed according to KHE and TA with and without Kasabach-Merritt phenomenon (KMP) respectively. SPSS 22.0 was used to compare the pooled rates among each therapy.Results:A total of 30 studies regarding the medication treatment of KHE and TA were identified in this meta-analysis. Analyzed medicines included glucocorticoid, vincristine, sirolimus, propranolol, combination therapy of vincristine and glucocorticoid. The pooled results indicated that when referring therapy on KHE and TA with KMP, the pooled response rate of combination therapy (98.34%) and sirolimus (96.43%) was higher than that of other therapies, and the difference was statistically significant. The pooled adverse events rate of sirolimus (5.53%) was relatively higher than other modalities, with no statistically significance. As for therapy on KHE and TA without KMP, sirolimus (94.23%) had higher pooled response rate than glucocorticoid (31.25%), vincristine (46.15%) and propranolol (22.86%), with statistically significant differences. The pooled adverse events rate of sirolimus was 23.81%.Conclusions:Our findings indicate that for KHE and TA with KMP, combination therapy (sirolimus + glucocorticoid) and vincristine have the best efficacy, while the adverse events rate of sirolimus is relatively high. For KHE and TA without KMP, sirolimus has the highest response rate, but there is also a risk of serious adverse events. Glucocorticoid and vincristine have comparable response rate, which both inferior to sirolimus.

Objective:To explore the application and choice of trunk perforator flaps in chest wound repair.Methods:From May 2015 to December 2019, clinical data of 14 patients using trunk perforator flap for chest soft tissue defect repair were reviewed. They were 8 males and 6 females, aged from 17 to 74 years. 5 patients had wound after scar surgery. 6 patients had benign lesions, and 3 patients had malignant tumors. The size of tissue defects ranged from 6.0 cm×8.0 cm-20.0 cm×21.0 cm. The perforator flaps were designed to repair different wounds. Preoperatively, the perforators are identified by ultrasound or CTA, and the flap scope and surgical approach are designed according to the perforator alignment and the perforated muscle point. Postoperatively, the survival of the flap was observed and followed up for at least 6 months.Results:All 17 flaps of 14 patients survived. The flap area was 7.0 cm × 8.0 cm-22.0 cm × 22.0 cm, and the length of the pedicle was 2.0-6.0 cm. All patients were followed up for 6 to 26 months with satisfied result . Recurrence occurred in one of the patients with thoracic dermatofibrosarcoma protuberans.Conclusions:The perforator artery of the trunk is densely and easy to obtain. The combined flaps could be used to repair large chest defects. The first-stage suture of the donor area, so it meets the organization transplantation principle of "the repair is perfect, the damage is small" .

Objective:To assess the efficacy and safety of common medication treatments on Kaposiform hemangioendothelioma (KHE) and tufted angioma (TA).Methods:PubMed, Embase, Web of Science, CNKI and Wanfang database were searched to find out the observational studies on medication treatment of KHE and TA. R-3.6.2 was used for calculate the pooled response rate and pooled adverse events rate. Meta analyses were performed according to KHE and TA with and without Kasabach-Merritt phenomenon (KMP) respectively. SPSS 22.0 was used to compare the pooled rates among each therapy.Results:A total of 30 studies regarding the medication treatment of KHE and TA were identified in this meta-analysis. Analyzed medicines included glucocorticoid, vincristine, sirolimus, propranolol, combination therapy of vincristine and glucocorticoid. The pooled results indicated that when referring therapy on KHE and TA with KMP, the pooled response rate of combination therapy (98.34%) and sirolimus (96.43%) was higher than that of other therapies, and the difference was statistically significant. The pooled adverse events rate of sirolimus (5.53%) was relatively higher than other modalities, with no statistically significance. As for therapy on KHE and TA without KMP, sirolimus (94.23%) had higher pooled response rate than glucocorticoid (31.25%), vincristine (46.15%) and propranolol (22.86%), with statistically significant differences. The pooled adverse events rate of sirolimus was 23.81%.Conclusions:Our findings indicate that for KHE and TA with KMP, combination therapy (sirolimus + glucocorticoid) and vincristine have the best efficacy, while the adverse events rate of sirolimus is relatively high. For KHE and TA without KMP, sirolimus has the highest response rate, but there is also a risk of serious adverse events. Glucocorticoid and vincristine have comparable response rate, which both inferior to sirolimus.

Objective:To explore the application and choice of trunk perforator flaps in chest wound repair.Methods:From May 2015 to December 2019, clinical data of 14 patients using trunk perforator flap for chest soft tissue defect repair were reviewed. They were 8 males and 6 females, aged from 17 to 74 years. 5 patients had wound after scar surgery. 6 patients had benign lesions, and 3 patients had malignant tumors. The size of tissue defects ranged from 6.0 cm×8.0 cm-20.0 cm×21.0 cm. The perforator flaps were designed to repair different wounds. Preoperatively, the perforators are identified by ultrasound or CTA, and the flap scope and surgical approach are designed according to the perforator alignment and the perforated muscle point. Postoperatively, the survival of the flap was observed and followed up for at least 6 months.Results:All 17 flaps of 14 patients survived. The flap area was 7.0 cm × 8.0 cm-22.0 cm × 22.0 cm, and the length of the pedicle was 2.0-6.0 cm. All patients were followed up for 6 to 26 months with satisfied result . Recurrence occurred in one of the patients with thoracic dermatofibrosarcoma protuberans.Conclusions:The perforator artery of the trunk is densely and easy to obtain. The combined flaps could be used to repair large chest defects. The first-stage suture of the donor area, so it meets the organization transplantation principle of "the repair is perfect, the damage is small" .

PURPOSE: The characteristic expression of DNA damage response proteins in familial breast cancers with BRCA1, BRCA2, or non-BRCA1/2 mutations has not been analyzed in Chinese patients. Our study aimed to assess the differential expression of microcephalin 1 (BRIT1), ATM serine/threonine kinase (ATM), checkpoint kinase 2 (CHEK2), BRCA1, RAD51 recombinase (RAD51), and poly (ADP-ribose) polymerase 1 (PARP-1) and establish the profile of Chinese familial breast cancers with different mutation status. METHODS: We constructed five tissue microarrays from 183 familial breast cancer patients (31 with BRCA1 mutations; 14 with BRCA2 mutations, and 138 with non-BRCA1/2 mutations). The DNA response and repair markers used for immunohistochemistry analysis included BRIT1, ATM, CHEK2, BRCA1, RAD51, and PARP-1. The expressions of these proteins were analyzed in BRCA1/2 mutated tumors. The association between pathologic characteristics with BRCA1/2 mutation status was also analyzed. RESULTS: In familial breast cancer patients, BRCA1 mutated tumors were more frequent with high nuclear grade, estrogen receptor/progesterone receptor/human epidermal growth factor receptor 2 negative, low Ki-67, and positive CK5/6. BRCA1 mutated tumors had lower CHEK2 and higher cytoplasmic BRIT1 expression than BRCA2 and non-BRCA1/2 mutation tumors. BRCA2-associated tumors showed higher CHEK2 and cytoplasmic RAD51 expression than those in other groups. Nuclear PARP-1 expression in BRCA1/2-associated tumors was significantly higher than in non-BRCA1/2 mutation tumors. Moreover, we found quite a few of negative PARP-1 expression cases in BRCA1/2 mutated groups. CONCLUSION: The clinicopathologic findings of BRCA1-associated Chinese familial breast cancers were similar to the results of other studies. Chinese familial breast cancer patients with BRCA1/2 mutations might have distinctive expression of different DNA damage response proteins. The reduced expression of PARP-1 in Chinese BRCA1/2 mutated breast cancer patients could influence the therapeutic outcome of PARP-1 inhibitors.
Asian Continental Ancestry Group* ; Breast Neoplasms* ; Breast* ; Checkpoint Kinase 2 ; Cytoplasm ; DNA Damage* ; DNA Repair ; DNA* ; Estrogens ; Genes, BRCA1 ; Genes, BRCA2 ; Humans ; Immunohistochemistry ; Phosphotransferases ; Rad51 Recombinase ; Receptor, Epidermal Growth Factor

Objective: To investigate whether N6-methyladenine DNA(6-mA DNA) modification is related to the occurrence of infantile hemangiomas (IH) at the epigenetic level. Methods: The genomic 6-mA DNA data were obtained by MeDIP and high-throughput sequencing. The 6-mA DNA methylation levels in 3 proliferative hemangioma specimens and adjacent skin tissues were compared by u-test. The functional differences of 6-mA modified genes were analyzed by GO analysis. Results: The level of 6-mA DNA modification in IH tissue was higher. The coverage of 6-mA Peaks in the genome was 0.037% in the tumor tissue, and the coverage of 6-mA Peaks in the surrounding skin tissue was 0.013% in the genome. The difference between the two groups was statistically significant (u=5999.87, P=0.00). The gene functions of differentially 6-mA modified genes in tumors were enriched in mesoderm development, stem cell differentiation, mesenchymal development, and cell cycle. These gene functions were closely related to the pathogenesis of IH. Conclusion Abnormal 6-mA DNA modification may be one of the pathogenesis of infantile hemangioma.

Objective: To describe the distribution and drug resistance of pathogens at hematology department of Jiangsu Province from 2014 to 2015 to provide reference for empirical anti-infection treatment. Methods: Pathogens were from hematology department of 26 tertiary hospitals in Jiangsu Province from 2014 to 2015. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or agar dilution method. Collection of drug susceptibility results and corresponding patient data were analyzed. Results: The separated pathogens amounted to 4 306. Gram-negative bacteria accounted for 64.26%, while the proportions of gram-positive bacteria and funguses were 26.99% and 8.75% respectively. Common gram-negative bacteria were Escherichia coli (20.48%) , Klebsiella pneumonia (15.40%) , Pseudomonas aeruginosa (8.50%) , Acinetobacter baumannii (5.04%) and Stenotropho-monas maltophilia (3.41%) respectively. CRE amounted to 123 (6.68%) . Common gram-positive bacteria were Staphylococcus aureus (4.92%) , Staphylococcus hominis (4.88%) and Staphylococcus epidermidis (4.71%) respectively. Candida albicans were the main fungus which accounted for 5.43%. The rates of Escherichia coli and Klebsiella pneumonia resistant to carbapenems were 3.5%-6.1% and 5.0%-6.3% respectively. The rates of Pseudomonas aeruginosa resistant to tobramycin and amikacin were 3.2% and 3.3% respectively. The resistant rates of Acinetobacter baumannii towards tobramycin and cefoperazone/sulbactam were both 19.2%. The rates of Stenotrophomonas maltophilia resistant to minocycline and sulfamethoxazole were 3.5% and 9.3% respectively. The rates of Staphylococcus aureus, Enterococcus faecium and Enterococcus faecalis resistant wards vancomycin were 0, 6.4% and 1.4% respectively; also, the rates of them resistant to linezolid were 1.2%, 0 and 1.6% respectively; in addition, the rates of them resistant to teicoplanin were 2.8%, 14.3% and 8.0% respectively. Furthermore, MRSA accounted for 39.15% (83/212) . Conclusions Pathogens were mainly gram-negative bacteria. CRE accounted for 6.68%. The rates of Escherichia coli and Klebsiella pneumonia resistant to carbapenems were lower compared with other antibacterial agents. The rates of gram-positive bacteria resistant to vancomycin, linezolid and teicoplanin were still low. MRSA accounted for 39.15%.

Objective To evaluate the clinical efficacy and safety of recipient-site pre-expansion via tumescent anesthesia in facial fat grafting .Methods Facial fat grafting was operated through the technology of recipient-site pre-expansion via tumescent anesthesia .The high-frequency ultrasonography was used to record the changes of soft tissue thickness , blood flow signal and transplant area in certain-points.Theresult were analyzed by SPSS 11.5.The survival rates of fat grafting were compared by Kruskal-Wallis H test.The area of soft tissue blood flow signals were analyzed by variance analysis .The patients' postoperative satisfaction rates and pain were evaluated by mean value .Results 14 cases were treated , with obvious improvement .Results of high frequency ultrasonography showed the median and quartile spacing of the soft tissue thickness was 70.1％(53.85％), 56.4％(51.95％), and 16.7％in the first month, third month and the sixth month after operation , separately.The difference between the three groups was statistically significant (H=1.4 ×108, P<0.05).The area of soft tissue blood flow signals was(3.97 ±0.84)‰,(2.21 ±0.55)‰,(6.89 ±2.39)‰,(14.61 ±4.66)‰,(9.39 ±3.94)‰, (15.87 ±3.02)‰before the operation, 24 hours, the first month, third month and the sixth month after operation,separately.The difference between the six groups were statistically significant (F=3.471, P <0.05).No complication occurred in the following-up period.Patients were satisfied with the pain relief. Conclusions It is safe and effective to apply recipient-site pre-expansion via tumescent anesthesia in fat grafting.

Objective To evaluate the clinical efficacy and safety of recipient-site pre-expansion via tumescent anesthesia in facial fat grafting .Methods Facial fat grafting was operated through the technology of recipient-site pre-expansion via tumescent anesthesia .The high-frequency ultrasonography was used to record the changes of soft tissue thickness , blood flow signal and transplant area in certain-points.Theresult were analyzed by SPSS 11.5.The survival rates of fat grafting were compared by Kruskal-Wallis H test.The area of soft tissue blood flow signals were analyzed by variance analysis .The patients' postoperative satisfaction rates and pain were evaluated by mean value .Results 14 cases were treated , with obvious improvement .Results of high frequency ultrasonography showed the median and quartile spacing of the soft tissue thickness was 70.1％(53.85％), 56.4％(51.95％), and 16.7％in the first month, third month and the sixth month after operation , separately.The difference between the three groups was statistically significant (H=1.4 ×108, P<0.05).The area of soft tissue blood flow signals was(3.97 ±0.84)‰,(2.21 ±0.55)‰,(6.89 ±2.39)‰,(14.61 ±4.66)‰,(9.39 ±3.94)‰, (15.87 ±3.02)‰before the operation, 24 hours, the first month, third month and the sixth month after operation,separately.The difference between the six groups were statistically significant (F=3.471, P <0.05).No complication occurred in the following-up period.Patients were satisfied with the pain relief. Conclusions It is safe and effective to apply recipient-site pre-expansion via tumescent anesthesia in fat grafting.