Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Treatment of large segmental defects of long bone has always been a major challenge for the medical community. At present, the bone defects have been treated by autologous or allogeneic bone grafting, free vascularized fibula grafting, Masquelet technique, Ilizarov technique, cylindrical mesh technique and other methods. However, these technical steps are relatively complicated and their cycles are long so that it is difficult to repair the bone defects immediately after surgery and carry out early weight bearing and functional exercise. In recent years, customized 3D printed microporous titanium (tantalum) prostheses have provided a safe and feasible treatment method for repair of these large bone defects. 3D printed microporous titanium (tantalum) prosthesis can accurately reconstruct the anatomical shape of a bone defect, and simultaneously provide stable biomechanical support for the overall bone structure by its mechanical strength of titanium alloy, demonstrating obvious advantages in the repair and regeneration of bone defects. This paper reviewed the characteristics, design requirements, mechanical properties, vascularization challenges and clinical applications of 3D printed microporous titanium (tantalum) prosthesis, and looked ahead to the future development of this new 3D printed prosthesis.

Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Treatment of large segmental defects of long bone has always been a major challenge for the medical community. At present, the bone defects have been treated by autologous or allogeneic bone grafting, free vascularized fibula grafting, Masquelet technique, Ilizarov technique, cylindrical mesh technique and other methods. However, these technical steps are relatively complicated and their cycles are long so that it is difficult to repair the bone defects immediately after surgery and carry out early weight bearing and functional exercise. In recent years, customized 3D printed microporous titanium (tantalum) prostheses have provided a safe and feasible treatment method for repair of these large bone defects. 3D printed microporous titanium (tantalum) prosthesis can accurately reconstruct the anatomical shape of a bone defect, and simultaneously provide stable biomechanical support for the overall bone structure by its mechanical strength of titanium alloy, demonstrating obvious advantages in the repair and regeneration of bone defects. This paper reviewed the characteristics, design requirements, mechanical properties, vascularization challenges and clinical applications of 3D printed microporous titanium (tantalum) prosthesis, and looked ahead to the future development of this new 3D printed prosthesis.

Objective:To explore the clinical phenotypic features and genetic variation characteristics of children with epilepsy-aphasia spectrum due to GRIN2A gene variants confirmed by second-generation sequencing. Methods:The clinical data of 5 children with epilepsy-aphasia spectrum with epileptic onset diagnosed in the Department of Neurology, Children′s Hospital Affiliated to Zhengzhou University, from February 2019 to November 2022 were retrospectively analyzed. Whole-exome genome sequencing of the probands using a second-generation sequencing method confirmed that all 5 cases were children with the GRIN2A gene variant. The characteristics of the GRIN2A gene variants were analyzed. Results:Among the 5 children diagnosed with epileptic aphasia spectrum due to GRIN2A gene variants, the male-to-female ratio was 4∶1, and the age range of onset was 1.5-4.4 years. The clinical phenotype included seizures in all cases, language and intellectual developmental deficits in 4 cases, and attention deficit hyperactivity disorder in 3 cases. The seizures were manifested as focal seizures or secondary generalized seizures, and were effectively controlled with antiepileptic drugs. Among the 5 children, gene variant of case 1 was originated from a paternal heterozygous variant, and cases 2-5 had de novo variants, which were c.2107C>T (p.Gln703 *) nonsense variant, c.2284G>A (p.Gly762Arg) missense variant, c.2197del (p.Ala733Glnfs *3) shifted coding variant, c.2511G>A (p.Trp837 *) nonsense variant, and c.1651+1G>C shear site variant, respectively. None of the 5 loci were reported in the literature. Conclusions:Epilepsy-aphasia spectrum is an epilepsy syndrome with a complex onset, and may have different phenotypes at different genetic variant loci, with focal seizures or secondary generalized seizures, which can be effectively controlled with anti-seizure medication. The GRIN2A gene variant is the genetic etiology of the epileptic aphasia spectrum.

Traditional Chinese medicine has the characteristics of multiple components， pathways， and targets in the treatment of fracture healing， and has good therapeutic advantages and potential for fractures with complex pathological mechanisms. Based on this， the author summarized the mechanism of promoting fracture healing by the monomer components and compound formulas of traditional Chinese medicine and found that visfatin A， puerarin， and others can activate the mitogen-activated protein kinase （MAPK） signaling pathway； Xugudan， Guben zenggu formula and others can activate bone morphogenetic protein （BMP） signaling pathway； baicalin， Achyranthes bidentata polysaccharides and others can activate Wnt/β -catenin signaling pathway； apigenin， notoginsenoside and others can activate receptor activator of nuclear factor-κB （NF-κB）/receptor activator of NF-κB ligand/osteoprotegerin （RANK/RANKL/OPG） signaling pathway； Compound huoxue jiegu capsule， Jiangu granule and others can inhibit phosphoinositide 3-kinase/protein kinase B （PI3K/AKT） signaling pathway； icariin can activate Notch signaling pathway； Taohong siwu decoction， crocin and others can activate Hippo signaling pathway； jujuboside A and osthole can inhibit NF-κB signaling pathway， and thus promote fracture healing.

Bilirubin has good anti-inflammatory， antioxidant and immunomodulatory effects， but its poor water solubility and low bioavailability greatly limit its clinical application. Researchers have developed bilirubin into various nanoparticles， which effectively eliminate the limitation of low solubility of bilirubin with the advantage of dosage form， so that they can maximize its pharmacological activities such as anti-inflammatory， anti-oxidation and immune regulation. Bilirubin nanoparticles have great application potential in a variety of gastrointestinal diseases， liver and kidney diseases， skin diseases， autoimmune diseases， islet transplantation and targeted therapy of tumors （both as a direct anti-tumor drug and as a drug delivery system）. The study of bilirubin nanoparticles will promote the clinical application of bilirubin and the development of related new drugs.

【Objective】 To evaluate the safety and efficacy of long-term indwelling of Allium ureteral stent in the treatment of ureteral stricture. 【Methods】 The clinical data of patients who underwent endoscopic Allium ureteral stent implantation for ureteral stricture in our hospital during Aug.2020 and Dec.2022 were retrospectively analyzed, and the surgical conditions and adverse events were recorded. The data of serum creatinine, blood urea nitrogen, glomerular filtration rate (GFR) and renal pelvis width under ultrasound were compared before surgery and 1, 3, 6 and 12 months after surgery. 【Results】 A total of 52 patients with ureteral stricture of 1.1 (0.7, 2.0)cm were included. All operations were successful. The operation time was 82.5 (70, 114)min, intraoperative blood loss 20 (10, 20)mL, and postoperative hospitalization stay 1 (1, 2) day. During the follow-up of (13.2±7.8) months, 14 patients had stent displacement, 5 had stone obstruction of stent tubes, 7 had occasional hematuria after movement, 9 had intermittent lumbar and abdominal pain, and 1 had recurrent urinary tract infection. The serum creatinine, blood urea nitrogen and renal pelvis width of 1 month, 3, 6 and 12 months after surgery were significantly decreased, while GFR was significantly increased. 【Conclusion】 Long-term indwelling of Allium ureteral stent is effective in the treatment of ureteral stricture, but the high incidence of stent displacement should arouse attention.

OBJECTIVE To investigate the protective effect and potential mechanism of Arisaema Cum Bile on acute liver injury induced by carbon tetrachloride （CCl4） in mice. METHODS Fifty mice were randomly divided into normal group， model group， positive control group （Biphenyl diester dropping pills， 150 mg/kg）， Arisaema Cum Bile low-dose and high-dose groups （0.78， 2.34 g/kg）， with 10 mice in each group. The mice in each group were given relevant medicine intragastrically， once a day， for 7 consecutive days. Two hours after the last administration， those groups were given intraperitoneal injection of 0.2% CCl4-olive oil solution to induce acute liver injury model except for normal group. Seventeen hours after intraperitoneal injection， the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， interleukin-6 （IL-6）， tumor necrosis factor-α（TNF- α）， and malondialdehyde （MDA）， superoxide dismutase （SOD） in liver tissue were measured with kit method. The hepatic index was detected. The pathological changes of liver tissue were observed by HE staining， and the degree of liver injury was scored quantitatively. The mRNA expressions of TNF-α and IL-6 in liver tissue were detected by real-time fluorescence quantitative PCR； the protein expressions of Janus kinase 2 （JAK2）， signal transducer and activator of transcription protein 3 （STAT3） and nuclear factor-κB p65 （NF-κB p65） in liver tissue were detected by Western blot assay. RESULTS Compared with normal group，the levels of ALT， AST， IL-6， TNF-α and MDA， the hepatic index were increased significantly （P＜0.05）， while the level of SOD was decreased significantly （P＜0.05）； the mRNA E-mail：qhwang668@sina.com expressions of IL-6 and TNF-α， and the protein expressions of JAK2， STAT3 and NF-κB p65 were up-regulated significantly （P＜0.05）； the pathological observation of liver tissue showed that the structure of hepatic cord was seriously disordered， there were many inflammatory cells infiltration of liver cells， and the liver injury score was significantly increased （P＜0.05）. Compared with model group， pathological changes and above indexes in mice were improved significantly in Arisaema Cum Bile low-dose and high-dose groups （P＜0.05）. CONCLUSIONS Arisaema Cum Bile has a protective effect on CCl4-induced acute liver injury in mice， which may be related to the inhibition of inflammatory response mediated by JAK2/STAT3/NF-κB signal pathway and antioxidant stress.

OBJECTIVE：To identify Panax notoginseng and its processed products . METHODS ：The fingerprint was established by HPLC. Using ginsenoside Rb 1 as reference ，HPLC fingerprints of 15 batches of P. notoginseng and its processed products were drawn and the similarity evaluation was conducted by using the Similarity Evaluation System for TCM Chromatographic Fingerprints（2012 edition）. The common peaks were confirmed by comparing with substance control. SPSS 21.0 and SIMCA 14.1 software were used to perform cluster analysis ，principal component analysis and orthogonal partial least squares-discriminant analysis；taking the variable importance projection （VIP）value greater than 1 as the standard ，the differential marker components causing the quality difference between P. notoginseng and its processed products were screened. IR fingerprints of P. notoginseng and its processed products were established by OMNIC 8.2.0 software，and the spectral similarity was evaluated ；double index sequence analysis was used to analyze absorption peaks of IR fingerprints of 15 batches of P. notoginseng and its processed products. RESULTS ：There were 16 common peaks in the fingerprints of 15 batches of P. notoginseng ， and the similarities were 0.911-1.000；there were 25 common peaks in the fingerprints of processed products ，and the similaritieswere 0.862-1.000. They had 12 identical common peaks ，and wang668@sina.com three of them were ident ified as sanchinoside R 1，ginsenoside Rg1 and ginsenoside Rb 1. Results of cluster analysi s showed that when the distance was 10，15 batches of P. notoginseng could be clustered into two categories ，SW1-SW5 into one category ，SH1-SH5 and SQ 1-SQ5 into one category ，ZW1-ZW5，ZH1-ZH5 and ZQ1-ZQ5 of 15 batches of processed products could be clustered into one category. When the distance was 5，15 batches of P. notoginseng could be clustered into three categories ，SW1-SW5 into one category ，SH2-SH5 and SQ 2 into one category ，SQ1， SQ3-SQ5 and SH 1 into one category. Fifteen batches of processed products could be clustered into two categories ，ZW1-ZW5 into one category ，ZH1-ZH5 and ZQ 1-ZQ5 into one category. The results of principal component analysis showed that the cumulative variance contribution rate of the first two principal components was 80.104％ . The results of orthogonal partial least squares-discriminant analysis showed that the VIP values of the five peaks were greater than 1，which were peak H ，peak G ，peak J，peak F （ginsenoside Rg 1）and peak I. The similarity of IR fingerprints of 15 batches of P. notoginseng and its processed products were 0.889 7-1.000 0 and 0.972 8-1.000 0；the common peak rates were 80％-100％，and the variation peak rates were 0-17.65％ and 0-18.75％，respectively. By comparing the wave numbers of absorption peaks ，it was found that there were differences between P. notoginseng at 3 440 and 1 450 cm－1 and processed products at 1 530 and 575 cm－1. CONCLUSIONS ：Established HPLC fingerprint and IR fingerprint have good similarity ，and could effectively distinguish P. notoginseng and its processed products. P. notoginseng and its processed products from different habitats have high common peak rate and low variation rate ，and their chemical components are different ；peak H ，peak G ，peak J ，ginsenoside Rg 1 and peak I are differential marker components causing the quality difference between P. notoginseng and processed products.