OBJECTIVE: To review the latest research advancements in surgical techniques for the treatment of limb lymphedema. METHODS: The relevant literature at home and abroad in recent years was extensively reviewed, and the research on the treatment of limb lymphedema by surgical techniques were summarized and analyzed. RESULTS: Lymphovenous anastomosis has demonstrated good effectiveness for early to mid-stage limb lymphedema, however its long-term effectiveness and applicability for late-stage limb lymphedema still require further validation. Autologous lymphatic/venous grafting has shown clinical feasibility in the treatment of secondary limb lymphedema. Research on tissue-engineered lymphatic scaffolds remains insufficient, primarily due to the complexity of lymphatic anatomical structures and the technical challenges involved. Nevertheless, its potential application is promising. Vascularized lymph node flap transplantation has shown significant effectiveness in treating limb lymphedema, particularly yielding good outcomes in upper limb cases. However, it can not guarantee a complete cure for the condition. Charles' operation is the most effective treatment option for patients with late-stage limb lymphedema, but its extensive incision and severe postoperative complications limit its application. Liposuction has the advantages such as minimal invasiveness, high safety, and repeatability. It is suitable for patients with late-stage limb lymphedema who have failed conservative treatment or developed adiposity. However, its effectiveness is limited in patients with significant limb fibrosis. CONCLUSION Current treatments for limb lymphedema require further improvement, and there is considerable debate regarding treatment strategies for different stages of the condition. Future high-quality, multi-system combined treatment approaches are anticipated to guide clinical practice.
Humans ; Lymphedema/surgery* ; Surgical Flaps/blood supply* ; Lymphatic Vessels/surgery* ; Anastomosis, Surgical/methods* ; Lymph Nodes/transplantation* ; Lipectomy/methods* ; Extremities/surgery* ; Treatment Outcome ; Tissue Engineering ; Tissue Scaffolds ; Veins/transplantation*

Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

Objective:To investigate whether neoadjuvant therapy can improve the prognosis of patients with pancreatic cancer.Methods:A retrospective case-control study analyzed data from the Surveillance, Epidemiology, and End Results (SEER) database on 12, 103 patients who underwent surgical treatment between January 1, 2010, and December 31, 2021. Patients were divided into the neoadjuvant therapy group ( n=3 276) and the upfront surgery group ( n=8 827) based on whether they received neoadjuvant treatment. The neoadjuvant therapy group included 2 342 patients receiving neoadjuvant chemotherapy and 934 patients receiving neoadjuvant chemoradiotherapy. The upfront surgery group consisted of 4 335 patients receiving adjuvant chemotherapy, 1 987 patients receiving adjuvant chemoradiotherapy, 63 patients receiving adjuvant radiotherapy, and 2 442 patients undergoing surgery alone. Propensity score matching was used to eliminate group differences and create a cohort with no statistical differences in other clinicopathological features except for the grouping variable. Variables such as age, gender, tumor location, race, population of residence, tumor diameter, household income, TNM stage, and information on radiotherapy and chemotherapy were used for 1∶1 case matching. T stage, N stage, and the use of radiotherapy or chemotherapy were matched exactly. After matching, 1 182 patients were included in each group: the neoadjuvant therapy group contained 1 155 patients receiving neoadjuvant chemoradiotherapy and 27 receiving neoadjuvant chemotherapy, while the upfront surgery group comprised 848 patients receiving adjuvant chemotherapy and 334 receiving adjuvant chemoradiotherapy. TNM staging was reported according to the 7th edition of the AJCC guidelines. The primary outcome was overall survival. Measurement data with skewed distributions were expressed as M( Q1, Q3), and intergroup comparisons were conducted using the Wilcoxon rank-sum test. Categorical data were compared using the chi-square test or the Fisher′s exact test. The Log-rank test and subgroup analyses to assess interactions between neoadjuvant therapy and subgroup in COX regression models were used to compare survival benefits across variables. Landmark analysis was performed to create segmented survival curves, studying the impact of neoadjuvant therapy on prognosis during different follow-up periods. Results:The neoadjuvant therapy group had a higher proportion of T 4 tumor involving celiac axis, superior mesenteric artery, and/or common hepatic artery compared to the upfront surgery group (14.7% vs 2.8%, P<0.001). Additionally, significant differences were observed between groups in terms of race, location, population of residence, age, tumor diameter, tumor stage, and adjuvant therapy regimen ( P<0.05). The median overall survival time in the neoadjuvant therapy group was 30 months, compared to 22 months in the upfront surgery group ( P<0.001). In the neoadjuvant therapy group, the median survival was 30 months for both neoadjuvant chemotherapy and chemoradiotherapy patients; in the upfront surgery group, it was 26 months for both adjuvant chemotherapy and chemoradiotherapy patients, 17 months for adjuvant radiotherapy patients, and 12 months for surgery-only patients. After propensity score matching, there were no differences in the distribution of clinical characteristics between groups ( P>0.05), and all patients in the matched cohort had received chemotherapy. The matched neoadjuvant therapy group had a longer median overall survival compared to the upfront surgery group (30 months vs 27 months, P<0.001). Subgroup interaction analysis revealed that T stage had a significant interaction with neoadjuvant therapy, both before (T 4 stage: HR=0.382, 95% CI: 0.319-0.458; T 2-T 3 stages: HR=0.696, 95% CI: 0.656-0.738; T 1 stage: HR=1.199, 95% CI: 0.867-1.657; interaction P<0.001) and after matching (T 4 stage: HR=0.581, 95% CI: 0.414-0.814; T 2-T 3 stages: HR=0.827, 95% CI: 0.734-0.931; T 1 stage: HR=1.320, 95% CI: 0.716-2.433; interaction P=0.043). Subgroup interaction analysis indicated that T 1 patients did not benefit from neoadjuvant therapy; survival curves plotted for matched T 1 patients showed no difference in survival between the neoadjuvant therapy group and the upfront surgery group ( P=0.323). Conversely, non-T 1 (T 2-T 4) stage patients showed significant survival benefits in both unmatched and matched cohorts ( P<0.001). Landmark analysis showing that the survival benefits occurred mainly in the early postoperative period of up to 3 years ( P<0.001), but there was no difference in overall survival between the neoadjuvant therapy group and the upfront surgery group of >3 years ( P>0.05). Patients with Arterial invasion (T 4 stage compared to T 1-T 3 stages) showed a similarly significant interaction with the benefit of neoadjuvant therapy in both the pre-matching cohort (interaction P<0.001) and the post-matching cohort (interaction P=0.037). Patients with T 4 stage disease in the neoadjuvant therapy group had longer overall survival compared to the upfront surgery group (median overall survival in pre-matching cohort: 30 months vs 13 months, P<0.001; median overall survival in post-matching cohort: 28 months vs 18 months, P=0.001). Among T 4 stage patients in the post-matching cohort, neoadjuvant therapy provided significant survival benefits during the early postoperative period of up to 3 years ( P=0.001). However, there was no difference in overall survival between the neoadjuvant therapy group and the direct surgery group beyond 3 years( P=0.729). Conclusions:The prognosis in the neoadjuvant therapy group was better than in the upfront surgery group. Propensity score matching and subgroup interaction analysis showed that non-T 1 and T 4 stage patients benefited more from neoadjuvant therapy, with benefits mainly seen in the early postoperative period (≤3 years).

BACKGROUND:Scaffold materials serve as platforms that provide space and structure,playing a crucial role in the regeneration of cartilage tissue.Scholars from around the world are exploring different approaches to fabricate more ideal scaffold materials. OBJECTIVE:To review the design principles and preparation methods of cartilage scaffolds,and to further explore the advantages and limitations of various preparation methods. METHODS:Literature searches were conducted on the databases of CNKI,WanFang Data,PubMed,and FMRS from 1998 to 2023.The search terms were"cartilage repair,cartilage tissue engineering,cartilage scaffold materials,preparation"in Chinese and English.A total of 57 articles were ultimately reviewed. RESULTS AND CONCLUSION:(1)The articular cartilage has a unique structure and limited self-repair capacity after injury.Even if self-repair occurs,the newly formed cartilage is typically fibrocartilage,which is far inferior to normal articular cartilage in terms of structure and mechanical properties.It is difficult to maintain normal function and often leads to degenerative changes.Currently,the design and fabrication of scaffold materials for cartilage repair need to consider the following aspects:biocompatibility and biodegradability,suitable pore structure and porosity,appropriate mechanical properties,and bioactivity.(2)Research on the preparation of cartilage scaffolds has made significant progress,continuously introducing new preparation methods and optimization strategies.These methods have their advantages and disadvantages,providing more possibilities for customized preparation and functional design of cartilage scaffolds according to specific requirements.

Objective To explore the relationship between alterations of neural network plasticity and spatial learning and memory functions in mouse models with depression-like behaviors.Methods C57Thy1-YFP/GAD67-GFP mice were randomized into control group(with no treatment)and chronic unpredictable mild stress(CUMS)group(n=15)subjected to CUMS for 8 weeks.Depression-like behaviors of the mice were assessed using sucrose preference test,open field test,and forced swimming test,and their spatial learning and memory abilities were evaluated using Morris water maze test.The changes in the firing patterns of different neuronal subtypes were detected in the central nucleus of the amygdala(CeA)and the prelimbic cortex(PrL)using whole-cell patch-clamp technique.Results Compared with the control mice,CUMS mice showed significantly decreased sucrose preference,total distance moved,number of grid-crossings,entries into the central area,and time spent in the central area in the open field test(P<0.01).In the forced swimming test,CUMS mice exhibited obviously shortened time of struggling,swimming,and climbing with increased immobility time.In Morris water maze test,CUMS mice showed significantly increased escape latency and path length,decreased percentage of distance and swimming time within the target quadrant,and increased first entry latency into the target zone and swimming time within the opposite quadrant.Exposure to CUMS resulted in significantly enhanced energy barrier and increased absolute refractory period and inter-spike interval of glutamatergic neurons in the CeA and GABAergic neurons in the PrL,while the opposite changes were observed in GABAergic neurons in the CeA and glutamatergic neurons in the PrL.Conclusion CUMS-induced depression may lead to plastic changes in the excitatory and inhibitory neuronal networks within the CeA and PrL to cause impairment of spatial learning and memory abilities in mice.

Objective To explore the relationship between alterations of neural network plasticity and spatial learning and memory functions in mouse models with depression-like behaviors.Methods C57Thy1-YFP/GAD67-GFP mice were randomized into control group(with no treatment)and chronic unpredictable mild stress(CUMS)group(n=15)subjected to CUMS for 8 weeks.Depression-like behaviors of the mice were assessed using sucrose preference test,open field test,and forced swimming test,and their spatial learning and memory abilities were evaluated using Morris water maze test.The changes in the firing patterns of different neuronal subtypes were detected in the central nucleus of the amygdala(CeA)and the prelimbic cortex(PrL)using whole-cell patch-clamp technique.Results Compared with the control mice,CUMS mice showed significantly decreased sucrose preference,total distance moved,number of grid-crossings,entries into the central area,and time spent in the central area in the open field test(P<0.01).In the forced swimming test,CUMS mice exhibited obviously shortened time of struggling,swimming,and climbing with increased immobility time.In Morris water maze test,CUMS mice showed significantly increased escape latency and path length,decreased percentage of distance and swimming time within the target quadrant,and increased first entry latency into the target zone and swimming time within the opposite quadrant.Exposure to CUMS resulted in significantly enhanced energy barrier and increased absolute refractory period and inter-spike interval of glutamatergic neurons in the CeA and GABAergic neurons in the PrL,while the opposite changes were observed in GABAergic neurons in the CeA and glutamatergic neurons in the PrL.Conclusion CUMS-induced depression may lead to plastic changes in the excitatory and inhibitory neuronal networks within the CeA and PrL to cause impairment of spatial learning and memory abilities in mice.

Adjuvant chemoradiotherapy,molecular targeted therapy,and immunotherapy are frequently employed to extend the survival of patients with advanced gastric cancer(GC).However,most of these treatments have toxic side effects,drug resistance,and limited improvements in survival and quality of life.Therefore,it is crucial to discover and develop new medications targeting GC that are highly effective and have minimal toxicity.In previous studies,the total terpene extract from the stem of Celastrus orbiculatus demonstrated anti-GC activity;however,the specific mechanism was unclear.Our research utilising co-immunoprecipitation-mass spectrometry(Co-IP-MS),polypyrimidine tract binding protein 1(ptbp1)clustered regularly interspaced short palindromic repeat-associated protein 9(Cas9)-knockout(KO)mouse model,tissue microarray,and functional experiments suggests that alpha actinin-4(ACTN4)could be a significant biomarker of GC.PTBP1 influences actin cytoskeleton restructuring in GC cells by interacting with ACTN4.Celastrus orbiculatus stem extract(COE)may directly target ACTN4 and affect the interaction between PTBP1 and ACTN4,thereby exerting anti-GC effects.

Objective:To preliminarily investigate the risk factors for distant metastasis and prognosis, and construct a prognostic nomogram in T 4 stage pancreatic cancer. Methods:A retrospective case-control study was conducted using data from the Surveillance, Epidemiology, and End Results (SEER) database for pancreatic patients from January 1, 2010, to December 31, 2021. Based on whether the tumor invaded the celiac axis, superior mesenteric artery, and/or common hepatic artery, 38 759 patients were divided into an arterial invasion group (T 4 stage, n=7 471) and a non-arterial invasion group (non-T 4 stage, n=31 288). Clinical and pathological data, including demographic characteristics, treatment information, and tumor data were collected. The primary outcome was overall survival. Categorical data were expressed as numbers (percentages), and intergroup comparisons were made using the chi-square test. Survival benefits were measured using the Log-Rank test. A multivariate logistic model was used to identify high-risk factors for metastasis in T 4 stage pancreatic cancer. Patients were randomly divided into training ( n=5 232) and validation ( n=2 239) sets at a 7∶3 ratio. A nomogram model was created based on independent prognostic factors from the multivariate Cox regression analysis, and the model′s predictive ability was evaluated using the C-index and calibration curves. Results:The overall metastasis rate in the arterial invasion group was higher than that in the non-arterial invasion group (32.8% vs 29.0%, P<0.001), with fewer patients showing no metastasis or single-organ metastasis (86.3% vs 89.7%, P<0.001) and higher rates of lung metastasis ( P<0.001), distant lymph node metastasis ( P<0.001), and other metastases excluding liver, lung, brain, bone, and distant lymph node metastases ( P<0.001). However, no significant difference was found between groups for liver, brain, or bone metastasis rates ( P>0.05). Surgical rates for T 4 stage patients were significantly lower than for non-T 4 stage patients (all patients: 10.7% vs 38.4%, P<0.001; M 0 stage patients: 15.0% vs 52.4%, P<0.001; M 1 stage patients: 2.1% vs 4.1%, P<0.001). Additionally, significant differences were observed in age, race, radiotherapy, chemotherapy, tumor location, tumor size, and tumor stage ( P<0.05). The median survival for patients with arterial invasion was 8 months, significantly lower than the 10-month median survival for non-arterial invasion patients ( P<0.001). The median survival for surgical patients with arterial invasion was 22 months, lower than the 24-month median for non-T 4 stage patients underwent surgery ( P<0.001) but significantly higher than for patients without surgery (T 4 stage patients without surgery: 8 months, P<0.001; non-T 4 stage patients without surgery: 6 months, P<0.001). For lymph node metastasis, patients with or without positive local lymph node metastasis had similar overall survival ( P>0.05). However, Patients with distant lymph node metastasis had significantly lower overall survival than that in patients without distant lymph node metastasis ( P<0.001). The multivariate logistic model indicated that tumor location in the body and tail ( OR=2.591, 95% CI: 2.343-2.867), positive regional lymph nodes ( OR=2.033, 95% CI: 1.836-2.252), and age <70 years old ( OR=1.183, 95% CI: 1.067-1.312) were risk factors for distant metastasis in arterial invasion patients. The multivariate Cox model showed that surgery ( HR=0.451, 95% CI: 0.405-0.503), radiotherapy ( HR=0.729, 95% CI: 0.677-0.784), chemotherapy ( HR=0.277, 95% CI: 0.258-0.297), tumor location in the body and tail ( HR=0.928, 95% CI: 0.874-0.985), and household income ≥$80, 000 ( HR=0.908, 95% CI: 0.853-0.968) were independent protective factors for prognosis in arterial invasion patients. Living in areas with a population ≤1 million ( HR=1.109, 95% CI: 1.044-1.178), age ≥70 years old ( HR=1.220, 95% CI: 1.150-1.296), larger tumor size (>2 cm but ≤4 cm: HR=1.124, 95% CI: 0.954-1.323; >4 cm: HR=1.310, 95% CI: 1.114-1.541), and having a metastatic burden (lung metastasis: HR=1.049, 95% CI: 0.869-1.267; distant lymph node metastasis: HR=1.179, 95% CI: 0.910-1.527; bone metastasis: HR=1.419, 95% CI: 0.854-2.359; brain or other metastasis: HR=1.519, 95% CI: 1.350-1.709; liver metastasis: HR=1.737, 95% CI: 1.600-1.886; two types of metastasis: HR=1.913, 95% CI: 1.689-2.168; three or more types: HR=2.436, 95% CI: 1.947-3.048) were independent risk factors for prognosis. The nomogram based on these prognostic factors had a C-index of 0.749 in the training set and 0.745 in the validation set; calibration curves in both sets were near the 45° line. Conclusions:High metastasis rates and low surgery rates are characteristic of pancreatic cancer with arterial invasion. Investigating the risk factors for distant metastasis and developing a prognostic nomogram incorporating metastatic burden hold significant clinical value for T 4 stage pancreatic cancer.

ObjectiveTo investigate the effect and mechanism of pachymic acid （PA） in Poria on the invasion and metastasis of renal carcinoma cells. MethodThe effect of PA （0， 20， 40， 80， 160 μmol·L^-1） on cell viability was detected by cell counting kit-8（CCK-8）， and the dose of PA was selected for subsequent experiments. The effect of PA （0， 20， 40， 80 μmol·L^-1） on cell proliferation was evaluated by colony formation assay. The effect of PA （0， 20， 40， 80 μmol·L^-1） on cell adhesion ability was observed by cell adhesion assay. The effect of PA （0， 20， 40， and 80 μmol·L^-1） on cell invasion and metastasis was investigated by Wound healing assay and Transwell invasion assay. The inhibitory effect of PA （0， 20， 40， 80 μmol·L^-1） on cell motility was further observed and verified by high-content imaging technology. The effects of PA （0， 20， 40， 80 μmol·L^-1） on the expression of matrix metalloproteinase （MMP）/tissue inhibitor of metalloproteinasas （TIMP） related to invasion and metastasis and Smads were detected by Western blot. ResultCCK-8 results showed that compared with the blank group， the PA groups showed decreased cell viability（P<0.01）， with the half-maximal inhibitory concentration （IC₅₀） of ACHN cells of 70.42 μmol·L^-1 at 24 h. Colony formation assay showed that the number of cell clonal groups in the PA groups was reduced compared with that in the blank group（P<0.01）. Cell adhesion assay showed that compared with the blank group， the PA groups displayed reduced cell adhesion（P<0.01）. Wound healing assay showed that the wound healing rate of cells in the PA groups was lower than that in the blank group （P<0.05，P<0.01）. Transwell invasion assay showed that compared with the blank group， the number of transmembrane cells in PA groups was reduced（P<0.01）. High-content imaging showed that the cumulative migration distance of cells in the PA groups was shorter than that in the blank group（P<0.01）. The results of Western blot showed that the protein expression of MMP-2 and MMP-9 in the PA groups decreased （P<0.01）， and TIMP-1 protein expression increased （P<0.01） compared with those in the blank group. In addition， compared with the blank group， the PA groups showed decreased protein expression of Smad2 and Smad3 （P<0.01）. ConclusionPA can inhibit the invasion and metastasis of renal carcinoma cells presumably through regulating the homeostasis of MMP/TIMP by Smad2/3.

Objective:To observe any effect of combining low-frequency transcranial magnetic stimulation (rTMS) with interactive virtual scenario training on the recovery of upper limb motor function after a stroke.Methods:Ninety stroke survivors were randomly divided into a pseudo-rTMS group, an rTMS group and a combination group, each of 30. In addition to basic medication, conventional rehabilitation and nursing care, the pseudo-rTMS, rTMS and combination groups received either sham rTMS treatment, 1Hz rTMS or virtual situational interaction along with 1Hz rTMS 5 days a week for 4 weeks. Before and after the 4 weeks their motor evoked potentials, cortical latency and central motor conduction time were measured, and surface electromyography was applied to the affected biceps brachii and triceps brachii. Meanwhile, the National Institutes of Health Stroke Scale, the Fugl-Meyer upper extremity assessment and the modified Barthel index were employed to assess the degree of neurological deficit, upper extremity motor function and ability in the activities of daily living (ADL).Results:After the 4-week intervention, a significant improvement was observed in all of the outcome measurements with all three groups. At that time the average scores of the rTMS group were significantly better than the pseudo-rTMS group′s averages but the average scores of the combination group were significantly better than those of either of the other two groups.Conclusion:Repeated application of low-frequency transcranial magnetic stimulation combined with virtual scenario interactive training can effectively improve the upper limb motor function and ADL performance of stroke survivors, and relieve the symptoms of neurological deficit. The combined therapy is worthy of application in clinical practice.