The key to the onset and progression of the functional impairments of orthopedic diseases is imba-lance of deficiency and excess， disharmony of movement and stillness， dislocation of sinews and bones， imbalance of rigidity and flexibility， and disharmony of body and spirit. Based on this， the "five views" treatment principle has been proposed for traditional Chinese medicine （TCM） orthopedic rehabilitation， which include the view of balancing deficiency and excess， combining movement and stillness， focusing on both sinews and bones， integrating rigidity and flexibility， and harmonizing body and spirit. Correspondingly， the "five views" collaborative rehabilitation clinical model is established， providing a reference for enhancing the systematization and targeting of TCM orthopedic rehabilitation.

Curzerene, a sesquiterpene compound isolated from Curcuma Radix, exhibits various therapeutic effects, such as anti-tumor and anti-hyperlipidemic properties. However, its neuroprotective effects have not yet been reported. This study focused on exploring the neuroprotective effect of curzerene and elucidating its potential mechanism by combining molecular biotechnology with multi-omics approaches. Curzerene was orally administered to LPS-induced depressive-like behaviors and cognitive impairment in mice for 14 days, and the related biochemical parameters were evaluated. The possible mechanisms were elucidated using qRT-PCR, Western Blot, immunofluorescence, untargeted metabolomics, GC-MS and 16S rDNA comprehensively. Curzerene ameliorated depression symptoms and cognitive impairment by increasing the preference for sucrose in SPT and the central area and total distance traveled in OFT, reducing the immobility time in TST and FST, as well as rising the spontaneous alternation ratio in Y maze. Multiple molecular biology techniques analyses indicated the ameliorative effect of curzerene via HMGB1/RAGE/ TLR4 pathway. Moreover, curzerene primarily regulates purine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, pyrimidine metabolism, etc. Furthermore, intervention increased the relative abundance of Parabacteroides, Clostridia_UCG-014_unclassified, and Rhodospirillales_unclassified, and enhanced the production of SCFAs. This work demonstrated that curzerene effectively protects against LPS-induced neurological damage, potentially by inhibiting the HMGB1/ RAGE/TLR4 pathway through the restoration of gut microbiota homeostasis, modulation of metabolites, and enhancement of SCFAs. In conclusion, this study offers new perspectives on the therapeutic possibilities of curzerene in mitigating depressive-like behaviors and cognitive impairment.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Objective To investigate the mechanism by which Xinqian Ganjie Decoction allevi-ates inflammatory responses in rat model of chronic rhinosinusitis by inhibiting the activation of the toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway.Methods Thirty-two 4-week-old healthy male Sprague-Dawley(SD)rats were randomly divided into model group(intragas-tric administration of 1.5 mL normal saline)and low-,medium-,and high-dose groups(intragastric administration of 0.75,1.50,and 3.00 mL/kg Xinqian Ganjie Decoction,respectively),with 8 rats in each group.Another 8 healthy rats were randomly selected as control group(intragastric adminis-tration of 1.5 mL normal saline).A rat model of chronic rhinosinusitis was established in the model group and the low-,medium-,and high-dose groups by placing expanded cotton pledgets in the nasal cavity and injecting Staphylococcus aureus suspension.After successful model establishment,rats in each group were treated by intragastric administration for 14 consecutive days.The number of snee-zes and nose scratches in each group was observed.Olfactory function changes in rats were assessed using an olfactory function test.Histopathological changes in nasal mucosa were detected by hema-toxylin-eosin(HE)staining.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in nasal mucosa were measured by enzyme-linked immunosor-bent assay(ELISA).The expression levels of proteins related to the TLR4/NF-κB signaling path-way were detected by western blot.Results Compared with the control group,rats in the model,low-,medium-,and high-dose groups showed increased number of sneezes and nose scratches.Compared with the model group,rats in the medium-and high-dose groups exhibited decreased number of sneezes and nose scratches.The differences among these groups were all statistically sig-nificant(P＜0.01).Before treatment,the time taken by rats to find food pellets in the control group was longer than that in the other groups(P＜0.01).After treatment,rats took longer to find food pellets in the model group compared with the control group,while rats in the low-,medium-,and high-dose groups took less time than the model group,with a decreasing trend in time as the dose increased.The differences among these groups were all statistically significant(P＜0.01).HE staining results showed that the nasal mucosa of rats in the control group was intact with no in-flammatory cell infiltration.Compared with the control group,the nasal mucosa of rats in the model group was significantly thickened with a large number of inflammatory cell infiltrations.Compared with the model group,the nasal mucosa thickening in rats of the medium-and high-dose groups was alleviated,with only a small amount of inflammatory cell infiltration observed.The levels of TNF-α,IL-1β,and IL-6 in the nasal mucosa of rats in the model group were higher than those in the control group.Compared with the model group,the levels of TNF-α,IL-1β,and IL-6 in the medium-and high-dose groups were decreased.The above differences among these groups were all statistically sig-nificant(P＜0.01).The expression levels of TLR4 and p-p65 NF-κB proteins in the nasal mucosa of rats in the model group were higher than those in the control group.Compared with the model group,the expression levels of TLR4 and p-p65 NF-κB in the medium-and high-dose groups were decreased.The differences among these groups were all statistically significant(P＜0.01).Con-clusion Xinqian Ganjie Decoction may reduce the release of TNF-α,IL-1β,and IL-6 by inhibiting the activation of the TLR4/NF-κB signaling pathway,thereby alleviating inflammatory responses and improving symptoms in a rat model of CRS.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Objective:To evaluate the value of ultrasonography in the differential diagnosis of cystic biliary atresia (CBA) and choledochal cyst (CC) in infants.Methods:CBA or CC children <3 months of age diagnosed by surgery in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from 2015 to 2023 were collected, and the differences in general conditions and ultrasound manifestations between CBA group and CC group were retrospectively analyzed.Results:Among the 55 children, 30 were in CBA group and 25 were in CC group. The mean length diameter and width diameter of cysts in the CBA group were about (1.45±1.05)cm and (1.04±0.73)cm, respectively, which were significantly smaller than those in the CC group (5.41±2.98)cm and (3.21±2.90)cm ( P<0.001). The incidence of fibrous plaque (50.0%, 15/30) and abnormal gallbladder morphology (73.3%, 22/30) in the CBA group was significantly higher than that in the CC group [4.0%(1/25) and 16.0%(4/25), respectively]. The incidence of intrahepatic biliary mud deposition (52.0%, 13/25) and intrahepatic biliary duct dilation (64.0%, 16/25) in the CC group was significantly higher than that in the CBA group [0, 3.3%(1/30), respectively]. Conclusions:There are some differences in ultrasonography between CBA and CC children. The length and width of cysts in children with CC are significantly larger than those in children with CBA, and biliary mud deposits or intrahepatic bile duct dilatation are seen in the cysts. The incidence of fibrous plaque and abnormal gallbladder morphology in CBA children is significantly higher than that in CC children.

Objective To quantify the setup errors for the different anatomical sites of patients who received intensity-modulated radiotherapy (IMRT) with linear accelerator on-board kilovolt fan beam CT(kV-FBCT) as non-isocenter IGRT and megavolt cone beam CT (MV-CBCT) as isocenter IGRT. Methods A retrospective analysis was performedon 70 patients who underwent radiotherapy, kV-FBCT, and/or MV-CBCT scans after each routine setup prior to IMRT. The average displacement (M), systematic error (Σ), and random error (б) at different treatment sites in the left-right, anterior-posterior, and cranial-caudal directions were calculated according to the individual displacements. The formula 2.5Σ+0.7б was used to estimate the PTV margin in respective direction. For each single patient, the root mean square in three directions was used as 3D displacement. Results A total of 1130 displacements were recorded in the 70 patients. The PTV margin was estimated to be 1.9-3.1 mm in head and neck cancer, 2.8-5.1 mm in thoracic cancer, 4.6-5.1 mm in breast cancer, 3.0-5.5 mm in upper abdominal cancer, and 3.5-6.8 mm in pelvic tumor. For the 3D mean displacements, the head and neck, thoracic, breast, upper abdominal, and pelvic cancer were 2.4±1.0, 4.0±1.6, 4.1±2.0, 4.6±2.1, and 4.6±2.1 mm, respectively. The average 3D displacement obtained by kV-FBCT and MV-CBCT were 4.1 and 3.4 mm, respectively (P=0.212). Conclusion The quantitative setup-error data can be obtained using linear accelerator on-board FBCT, and the non-isocenter IGRT induced set-up error cannot be negligible.

Ulcerative colitis (UC) is one of types of inflammatory bowel disease with high recurrence. Recent studies have highlighted that microbial dysbiosis as well as abnormal gut immunity are crucial factors that initiate a series of inflammatory responses in the UC. Modulating the gut microbiota-intestinal immunity loop has been suggested as one of key strategies for relieving UC. Many Chinese herbal medicines including some of single herb, herbal formulas and the derived constituents have been reported with protective effect against UC through modulating gut microbiome and intestinal immunity. Some clinical trials have shown promising results. This review thus focused on the current knowledge on using Chinese herbal medicines for treating UC from the mechanism aspects of regulating intestinal homeostasis involving microbiota and gut immunity. The existing clinical trials are also summarized.

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.

Enzyme-catalysis self-assembled oligopeptide hydrogel holds great interest in drug delivery, which has merits of biocompatibility, biodegradability and mild gelation conditions. However, its application for protein delivery is greatly limited by inevitable degradation of enzyme on the encapsulated proteins leading to loss of protein activity. Moreover, for the intracellularly acted proteins, cell membrane as a primary barrier hinders the transmembrane delivery of proteins. The internalized proteins also suffer from acidic and enzymatic degradation in endosomes and lysosomes. We herein develop a protease-manipulated hybrid nanogel/nanofiber hydrogel for localized delivery of intracellularly acted proteins. The embedded polymeric nanogels (CytoC/aNGs) preserve activity of cytochrome