Objective:To summarize the clinical characteristics of newly-developed lesions of early esophageal cancer and precancerous lesions after endoscopic submucosal dissection (ESD) , and to investigate the risk factors associated with metachronous multiple primary early esophageal cancers.Methods:A retrospective analysis was conducted on clinical data and postoperative follow-up results of 311 patients who underwent esophageal ESD at the Department of Gastroenterology, Taizhou People's Hospital, from January 2018 to January 2020. The incidence and interval of newly-developed lesions were documented, and the risk factors for metachronous multiple primary esophageal cancers were identified.Results:Among the 311 patients, 1 case (0.3%) experienced local recurrence, 2 patients (0.6%) had synchronous multiple primary early esophageal cancers, and 27 cases (8.7%) had metachronous multiple primary early esophageal cancers. Multivariate regression analysis showed that lesion length ( OR=5.728, 95% CI: 0.959-34.208, P<0.001), Lugol-voiding lesions (LVLs) with speckled distribution ( OR=6.574, 95% CI: 2.163-19.977, P<0.001), and high neutrophil-to-lymphocyte ratio (NLR) ( OR=3.72, 95% CI: 2.144-6.452, P<0.001) were independent risk factors for metachronous multiple primary early esophageal cancers. Conclusion:Incidence of metachronous multiple primary early esophageal cancers is elevated in patients with long lesions, LVLs exhibiting speckled distribution, and high NLR. Therefore, close follow-up is essential for patients displaying these identified risk factors.

Objective To compare the size discrepancy between ultrasonic and pathological measurement of solitary cN0M0 papillary thyroid microcarcinoma(PTMC),and to explore their correlation with lymph node metastasis.Methods From April 2021 to January 2022,234 patients with solitary cN0M0 PTMC who received thyroid lobectomy or total thyroidectomy in the Department of Thyroid and Breast Surgery of Nanjing University of Chinese Medicine,Affiliated Hospital of Integrated Traditional Chinese and Western Medicine were analyzed retrospectively.The size discrepancy between ultrasonic and pathological measurement were compared,and the risk factors of central lymph node metastasis were analyzed.Results The mean of maximum diameter of PTMC measured by ultrasound was 6.8(range 5.6 to 8.4)mm,which was significantly bigger than that measured by pathology 5.0(range 4.0 to 7.0)mm(P=0.000).Of them,37.2%of the tumor size measured by ultrasound is consistent with pathology,61.1%of the tumor size measured by ultrasound is bigger than that measured by pathology,and only 1.7%of the tumor size measured by ultrasound is smaller than that measured by pathology.There was a linear correlation between the diameter measured by ultrasound and pathology.And the regression equation can be expressed as:pathological diameter=0.799×ultrasonic diameter-0.221.In addition,28.6%patients had central lymph node metastasis.Multivariate Logistic regression analysis showed that the diameter measured by pathology is a risk factor for central lymph node metastasis in patients(OR=17.845,95%CI:2.507-127.025,P=0.004),and the cutoff value is 5.5 mm which corresponded to the diameter measured by ultrasound as 7.2 mm.Conclusions The sizes of solitary cN0M0 PTMC measured by ultrasound and pathology are different but also correlated.PMTC with pathological diameter>5.5 mm with its corresponding ultrasonic diameter as 7.2 mm indicated an increased risk of central lymph node metastasis.

【Objective】 To retrospectively analyze the serological and nucleic acid testing(NAT) data of voluntary blood donors from six blood banks in Tibet, in order to explore the positive impact of NAT on reducing the risk of infective transfusion in a regional scope. 【Methods】 From 2018 to 2022, 38 718 voluntary blood donors from blood centers of Tibet, Shannan, Shigatse, Naqu, Nyingchi and Ngari were tested for hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), human immunodeficiency virus antigen (HIV) and antibody (Ag/Ab1+2) serological determination by enzyme-linked immunosorbent assay (ELISA). At the same time, Haoyuan and Daan nucleic acid detection systems were used for the combined detection of HBV-DNA, HCV-RNA and HIV-RNA. The results of NAT of reactive ELISA samples were statistically analyzed. 【Results】 A total of 178 ELISA-/NAT+ samples were detected in Tibet over the past five years, including 170 HBV-DNA positive cases, 8 HCV-RNA positive cases, and 0 HIV-RNA positive cases, with the positive rate at 0.460%.The detection rate of 624 ELISA+/NAT+ samples was 1.61%.The age of blood donors with hepatitis B in Shigatse area was slightly higher than that in other areas, and the difference was statistically significant(P<0.05) . 【Conclusion】 The centralized detection of viral nucleic acid in Blood Center of Tibet Autonomous Region can effectively reduce the missed detection of transfusion transmitted diseases and guarantee the blood safety in the region.

Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.

Background:With the advancement of endoscopic techniques,the detection rate of duodenal lesions has increased,and the selection and curative effect of endoscopic resection of duodenal lesions need to be further explored.Aims:To evaluate the safety and efficacy of endoscopic treatment of duodenal tumors.Methods:A retrospective analysis was conducted on 32 patients with duodenal tumors treated with endoscopic resection from January 2017 to January 2022 at Taizhou People's Hospital Affiliated to Nanjing Medical University.The location,depth,treatment method,postoperative pathology,and complications of duodenal tumors were summarized.Results:Among the 32 patients,there were 21 males and 11 females.Two patients presented with melena,10 patients had abdominal pain,and 20 patients were asymptomatic.Twenty-eight tumors were located in descending part of duodenum,and 4 were located in duodenal bulb.One patient underwent endoscopic submucosal excavation,2 underwent cold snare polypectomy,12 underwent endoscopic submucosal dissection,and 17 underwent endoscopic mucosal resection.All lesions were completely resected.Among the 32 lesions,there were 21 tubular adenoma or tubulovillous adenoma,7 Brunner's gland adenoma,2 lipoma,1 neuroendocrine tumor,and 1 stromal tumor.One patient with duodenal tubulovillous adenoma had perforation during operation and was transferred to surgery for repair.The other patients did not have complications such as perforation and bleeding during operation.All patients had no recurrence after postoperative endoscopy.Conclusions:Endoscopic treatment for duodenal tumors is safe and effective,and has good clinical value.

Objective:To study the application of establishing WeChat group combined with PBL teaching mode in the practice of pediatric orthopaedics.Methods:The study was conducted among 36 postgraduates of "5+3" pediatrics and 22 postgraduates of orthopedic surgery of Batch 2012 who had practice in Department of Orthopaedics, Children's Hospital of Chongqing Medical University from September 2018 to August 2019, and they were randomized into control group and experimental group, with 29 students in each group. The control group was taught by traditional PBL mode, while the experimental group was taught by WeChat group combined with PBL mode. At the time of leaving the department, the two groups were assessed by theoretical knowledge, practical operation and questionnaire satisfaction, thus evaluating the teaching effects. The SPSS 23.0 was used to conduct t test and chi-square test. Results:The average scores of theoretical knowledge test (91.28±2.89) and practical operation test (87.44±2.94) in the experimental group were significantly higher than those in the control group (87.39±3.53) and (79.06±3.84), with statistically significant differences ( P<0.05). In the survey of teaching satisfaction, the experimental group[96.55%(27/29)] was significantly higher than the control group[61.90%(21/29)], with statistically significant differences ( P<0.05). Conclusion:The establishment of WeChat group and the combination of PBL teaching model can significantly improve the teaching effect and students' satisfaction in the practice of pediatric orthopaedics, which is worthy of further promotion and application.

Objective:To investigate the effects of different chimerism strategies and different immune ways on the two antigen-dominant regions of Xinjiang hemorrhagic fever virus (XHFV) glycoprotein.Methods:The 5' end was added or not added with interleukin-2 (IL-2) signal peptide and the general-purpose auxiliary T cell epitopes as different design strategies. GcⅠ and GcⅡ and the epitopes previously identified on GcⅠ (Gc 233-248, Gc 241-256 and Gc 281-296) were fused and constructed into the eukaryotic expression vector pVAX1 and the prokaryotic expression vector pET-28a. The recombinant prokaryotic plasmid transformed into E.coli BL21 was induced and purified, and the recombinant eukaryotes were extracted by indirect immunofluorescent assay. BALB/c mice were immunized by protein immunity, gene immunity, and DNA prime-protein boost immunity. The IgG antibody level was measured by ELISA. The immune effect was evaluated by the proliferation of T-lymphocytes and the content of cytokines in the spleen. Results:The results of double enzyme digestion and sequencing showed that eight recombinant plasmids were successfully constructed, and the recombinant eukaryotes were successfully expressed in vitro by fluorescence microscopy. After three times of immunization, the IgG level and the proliferation of T-lymphocytes in the spleen of mice in the experimental group were significantly higher than those in the control group ( P<0.01). The mass concentration test results of Th2 cytokines IL-4 and Th1 cytokines interferon-gamma (IFN-γ) revealed that the response of the DNA prime-protein boost immunity was biased to Th1. Conclusions:The multi-epitope chimeric vaccine of XHFV glycoprotein was successfully constructed, and the target antigen could be expressed effectively in vivo. The immune groups stimulated stronger humoral and cellular immune responses compared with the control group. Among them, the immune effect of pVAX1-ST(GcⅠe+GcⅡ) combined with recombinant protein r(GcⅠe+GcⅡ) was the best, and it is expected to be a new candidate vaccine for XHFV.

With the emergence of DNA nanotechnology in the 1980s, self-assembled DNA nanostructures have attracted considerable attention worldwide due to their inherent biocompatibility, unsurpassed programmability, and versatile functions. Especially promising nanostructures are tetrahedral framework nucleic acids (tFNAs), first proposed by Turberfield with the use of a one-step annealing approach. Benefiting from their various merits, such as simple synthesis, high reproducibility, structural stability, cellular internalization, tissue permeability, and editable functionality, tFNAs have been widely applied in the biomedical field as three-dimensional DNA nanomaterials. Surprisingly, tFNAs exhibit positive effects on cellular biological behaviors and tissue regeneration, which may be used to treat inflammatory and degenerative diseases. According to their intended application and carrying capacity, tFNAs could carry functional nucleic acids or therapeutic molecules through extended sequences, sticky-end hybridization, intercalation, and encapsulation based on the Watson and Crick principle. Additionally, dynamic tFNAs also have potential applications in controlled and targeted therapies. This review summarized the latest progress in pure/modified/dynamic tFNAs and demonstrated their regenerative medicine applications. These applications include promoting the regeneration of the bone, cartilage, nerve, skin, vasculature, or muscle and treating diseases such as bone defects, neurological disorders, joint-related inflammatory diseases, periodontitis, and immune diseases.
Nucleic Acids/chemistry* ; Regenerative Medicine ; Consensus ; Reproducibility of Results ; DNA/chemistry*

Objective:To investigate the med- to long-term clinical outcomes of open reduction and cannulated screw fixation via the modified Smith-Petersen approach for irreducible femoral neck fractures in young adults.Methods:The clinical data of 16 young adults were retrospectively studied who had been treated for irreducible femoral neck fractures by open reduction and cannulated screw fixation via the modified Smith-Petersen approach by the same surgeon from December 2009 to February 2015 at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital. There were 11 males and 5 females aged from 19 to 59 years (average, 42.9 years). All their fractures were closed Garden type Ⅳ, with high energy injury in 8 cases and low energy injury in the other 8 ones. The time from injury to surgery averaged 5.4 d (from 1 to 11 d). In all the patients, open reduction and cannulated screw fixation via the modified Smith-Petersen approach was conducted after ideal reduction had not been achieved by 3 continuous attempts in primary closed reduction in a traction bed. Recorded were the postoperative full weight-bearing time, return to work time, fracture union time, function of the affected hip and complications at the last follow-up in the patients.Results:The 16 patients were followed up for 6 to 12 years (average, 8.8 years). The postoperative full weight-bearing time averaged 12.7 weeks (from 2 to 16 weeks) in 16 patients; the return to work time averaged 15.0 weeks (from 3 to 20 weeks) and the fracture union time 3.5 months (from 3 to 6 months) in 15 patients while fracture nonunion happened in the other one. The HSS hip scores at the last follow-up averaged 90.3 points (from 62 to 98 points), yielding 12 excellent, 2 good, one fair and one poor cases. The one case with fracture nonunion underwent total hip arthroplasty (THA); of the 4 cases with avascular necrosis of the femoral head, 2 underwent THA; limited range of hip motion due to heterotopic ossification was noted in one case.Conclusion:Open reduction and cannulated screw fixation via the modified Smith-Petersen approach may lead to fine med- to long-term clinical outcomes in young adults whose femoral neck fracture cannot be reduced by closed reduction, because the modified Smith-Petersen approach allows fracture reduction under direct vision.

Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.