Collagen is a major structural protein in the matrix of animal cells and the most widely distributed and abundant functional protein in mammals. Collagen’s good biocompatibility, biodegradability and biological activity make it a very valuable biomaterial. According to the source of collagen, it can be broadly categorized into two types: one is animal collagen; the other is recombinant collagen. Animal collagen is mainly extracted and purified from animal connective tissues by chemical methods, such as acid, alkali and enzyme methods, etc. Recombinant collagen refers to collagen produced by gene splicing technology, where the amino acid sequence is first designed and improved according to one’s own needs, and the gene sequence of improved recombinant collagen is highly consistent with that of human beings, and then the designed gene sequence is cloned into the appropriate vector, and then transferred to the appropriate expression vector. The designed gene sequence is cloned into a suitable vector, and then transferred to a suitable expression system for full expression, and finally the target protein is obtained by extraction and purification technology. Recombinant collagen has excellent histocompatibility and water solubility, can be directly absorbed by the human body and participate in the construction of collagen, remodeling of the extracellular matrix, cell growth, wound healing and site filling, etc., which has demonstrated significant effects, and has become the focus of the development of modern biomedical materials. This paper firstly elaborates the structure, type, and tissue distribution of human collagen, as well as the associated genetic diseases of different types of collagen, then introduces the specific process of producing animal source collagen and recombinant collagen, explains the advantages of recombinant collagen production method, and then introduces the various systems of expressing recombinant collagen, as well as their advantages and disadvantages, and finally briefly introduces the application of animal collagen, focusing on the use of animal collagen in the development of biopharmaceutical materials. In terms of application, it focuses on the use of animal disease models exploring the application effects of recombinant collagen in wound hemostasis, wound repair, corneal therapy, female pelvic floor dysfunction (FPFD), vaginal atrophy (VA) and vaginal dryness, thin endometritis (TE), chronic endometritis (CE), bone tissue regeneration in vivo, cardiovascular diseases, breast cancer (BC) and anti-aging. The mechanism of action of recombinant collagen in the treatment of FPFD and CE was introduced, and the clinical application and curative effect of recombinant collagen in skin burn, skin wound, dermatitis, acne and menopausal urogenital syndrome (GSM) were summarized. From the exploratory studies and clinical applications, it is evident that recombinant collagen has demonstrated surprising effects in the treatment of all types of diseases, such as reducing inflammation, promoting cell proliferation, migration and adhesion, increasing collagen deposition, and remodeling the extracellular matrix. At the end of the review, the challenges faced by recombinant collagen are summarized: to develop new recombinant collagen types and dosage forms, to explore the mechanism of action of recombinant collagen, and to provide an outlook for the future development and application of recombinant collagen.

OBJECTIVE: To evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

OBJECTIVES: To identify risk factors contributing to prolonged postoperative length of stay (LOS) in very elderly patients following hip fracture surgery, with a focus on postoperative complications and the impact of different anesthesia approaches. METHODS: This retrospective single-center cohort study enrolled patients aged 90 years or older who underwent hip fracture surgery at Peking Union Medical College Hospital between January 31, 2013 and December 31, 2023. Relevant perioperative data were collected. The primary outcome was postoperative LOS, and the study cohort was divided into two groups: postoperative LOS ≤ 7 days and LOS > 7 days. Logistic regression was performed to identify factors related to prolonged postoperative LOS. RESULTS: A total of 155 patients were included. The average age was 92.7 ± 2.6 years. There were 73 (47%) patients with postoperative LOS > 7 days. Postoperative pneumonia was the only factor associated with a prolonged postoperative LOS (OR = 2.12, 95% CI [1.09, 4.16], P = 0.028). Neither the type of anesthesia (regional vs. general anesthesia, OR = 1.00, 95% CI [0.53, 1.90], P = 0.993) nor the method of airway management (laryngeal mask ventilation vs. spontaneous breathing, OR = 1.46, 95% CI [0.58, 3.76], P = 0.424; endotracheal intubation vs. spontaneous breathing, OR = 0.82, 95% CI [0.39, 1.69], P = 0.592) showed a significant association with a prolonged postoperative LOS. Preoperative chronic obstructive pulmonary disease (OR = 2.78, 95% CI [1.05, 7.65], P = 0.040) and preoperative neutrophil count (OR = 1.13, 95% CI [1.01, 1.26], P = 0.029) were both significantly associated with the occurrence of postoperative pneumonia, while anesthesia type and airway management method were not. CONCLUSIONS Postoperative pneumonia was associated with prolonged postoperative LOS in very elderly patients undergoing hip fracture surgery, whereas anesthesia types and airway management methods show no association with prolonged postoperative LOS or postoperative pneumonia. Preoperative comorbidities, especially respiratory conditions and systemic inflammation, potentially play a substantial role in postoperative recovery.
Humans ; Hip Fractures/surgery* ; Aged, 80 and over ; Risk Factors ; Length of Stay ; Female ; Male ; Retrospective Studies ; Postoperative Complications/etiology*

Objective:To investigate the clinical application value of artificial intelligence(AI)-based bone marrow cell recognition and analysis system in the diagnosis of hematological diseases.Methods:The bone marrow smears of hematological patients who were admitted to The Second Hospital of Shanxi Medical University from 2018 to 2020 were retrospectively analyzed.A total of 115 bone marrow smears with clear diagnosis and typical cell morphology characteristics were selected,including 20 cases of immune thrombocytopenia(ITP),11 cases of iron deficiency anemia(IDA),17 cases of megaloblastic anemia(MA),20 cases of chronic myeloid leukemia(CML),17 cases of acute lymphoblastic leukemia(ALL),23 cases of acute promyelocytic leukemia(APL),and 7 cases of acute myeloid leukemia unclassified(AML-M2).The samples were analyzed by manual microscopic examination,AI automatic recognition,and manual correction after AI recognition.Results:The images captured by the AI device were clear,and the cell morphological structures were distinct.The average experimental diagnostic efficiency parameters of the bone marrow nucleated cells classified in this system were calculated.The sensitivity was 74.90％,specificity was 99.03％,and accuracy was 98.29％.In the comparison between the AI recognition group and the manual examination group,the data of IDA,ITP,MA,and CML diseases were all greater than 0.85 in ICC correlation coefficient,with excellent consistency;the data of APL,AML-M2,and ALL three diseases were between 0.6 and 0.85 in ICC correlation coefficient,with moderate consistency.However,after manual review and correction,the ICC correlation coefficient between the data of the AI correction group and the data from the manual examination group was greatly improved.Conclusion:The AI bone marrow cell recognition and analysis system has the characteristics of high accuracy,high specificity,good sensitivity and fast detection.When used in combination with manual review,it can improve the detection efficiency of bone marrow cells morphological analysis and meet the needs of clinical work.

Objective:To investigate the early warning value of transcutaneous oxygen pressure and transcutaneous carbon dioxide pressure (TcPO 2/TcPCO 2) monitoring in patients with pressure injury in ICU. Methods:A prospective clinical observation study was conducted. Patients were selected in the department of critical care medicine of Shenzhen Hospital Affiliated to University of Chinese Academy of Sciences from December 2020 to June 2022. The general information of all patients were recorded. The data of TcPO 2 and TcPCO 2 were monitored respectively in left lying position, supine position and right lying position. According to the Braden score of patients, they were divided into low risk group and high risk group, and the monitoring results of TcPO 2/TcPCO 2 were analyzed. Results:The study included a total of 80 patients, with 46 patients in the low risk group and 34 patients in the high risk group. There were no significant differences in gender and age between the two groups (both P>0.05). There were significant differences in the Braden score and the incidence of pressure injury between the two groups (both P<0.05). There were statistically significant differences in TcPO 2 at various time points in the left lying position, supine position and right lying position between the low risk and the high risk groups (all P<0.001). Except for the 15th minute of the left lying position and right lying position, there were statistically significant differences in TcPCO2 at all other time points in the supine and right lying positions between two groups (all P<0.05). There were statistically significant differences in TcPO 2/TcPCO 2 values at various time points in different positions between two groups (all P<0.001). In supine position, TcPO 2 and TcPO 2/TcPCO 2 were positively correlated with the Braden score ( r=0.680, 0.741). TcPCO 2 was negatively correlated with the Braden score ( r=-0.771). The Braden score, TcPO 2, TcPCO 2 and TcPO 2/TcPCO 2 all have moderate diagnostic value. The cut-off values were 12.50, 41.48 mmHg, 52.29 mmHg and 0.91, respectively. And the AUC were 0.899, 0.727, 0.816 and 0.719, respectively. However, there were no significant differences in AUC between the TcPO 2, TcPCO 2, TcPO 2/TcPCO 2 and Braden score (all P>0.05). Conclusions:TcPO 2, TcPCO 2 and TcPO 2/TcPCO 2 values all have higher diagnostic value about pressure injury for patients in ICU. TcPO 2/TcPCO 2 monitoring has early risk warning value for the occurrence of pressure injury in ICU, which is worthy of clinical promotion.

Objective To explore the regulatory role of transient receptor potential channel 6(TRPC6)on podocyte autophagy under the influence of homocysteine(Hcy)in mouse kidney.Methods Mouse renal podocytes were divided into control group and Hcy groups(stimulated by Hcy at 40,60,80 and 100 μmol/L for 48 h).The level of TRPC6 mRNA was assessed using quantitative reverse transcription polymerase chain reaction(qRT-PCR)to identify the optimal Hcy concentration for subsequent experiments.Western blotting was employed to evaluate the expression levels of autophagy-related proteins LC3 Ⅱ and p62,as well as the expression levels of podocyte structural proteins Nephrin and Podocin.The expression levels of TRPC6 mRNA and protein in both groups were determined using qRT-PCR,Western blotting and immunofluorescence.Transfections of cells with TRPC6 overexpression or interference were set as follows:(1)control group(untreated),negative control group of TRPC6 overexpression,and TRPC6 overexpression group;(2)control group(untreated),negative control group of TRPC6 interference,and TRPC6 interference group(si-1,si-2,si-3).The expression level of TRPC6 was detected using qRT-PCR.The cells after overexpressing or interfering of TRPC6 were further set as follows:(1)control group(untreated),Hcy group(80 μmol/L Hcy added),TRPC6 overexpression control+Hcy group,TRPC6 overexpression+Hcy group;(2)control group(untreated),Hcy group,TRPC6 interference control+Hcy group,and TRPC6 interference+Hcy group.The expression levels of p62,LC3 Ⅱ,and TRPC6 proteins were detected using Western blotting.Results qRT-PCR detection results showed that compared with control group,the expression level of TRPC6 mRNA in Hcy group increased with the increase of Hcy concentration,with the highest expression level observed at 80 μmol/L Hcy.Therefore,80 μmol/L Hcy was selected as the optimal concentration for intervention.At this time,the expression level of autophagy-related protein LC3 Ⅱ increased,and the expression level of p62 decreased(P<0.05).Western blotting results showed that compared with control group,the expression levels of podocyte-related proteins Nephrin and Podocin in Hcy group were significantly decreased(P<0.05).qRT-PCR results showed that compared with control group,the expression level of TRPC6 mRNA in Hcy group was significantly increased(P<0.05).Compared with negative control group for TRPC6 overexpression,both mRNA and protein expression levels of TRPC6 in TRPC6 overexpression group were significantly higher(P<0.05).Compared with negative control group for TRPC6 interference,both mRNA and protein expression levels of TRPC6 in TRPC6 interference group were significantly decreased(P<0.05).Western blotting results showed that compared with negative control group for TRPC6 overexpression,the expression level of autophagy-related protein LC3 Ⅱ in TRPC6 overexpression+Hcy group was significantly increased,and the expression level of p62 was significantly decreased(P<0.05).Compared with TRPC6 negative control+Hcy group for TRPC6 interference+Hcy,the expression level of autophagy-related protein LC3 Ⅱ in TRPC6 interference+Hcy group was significantly decreased,and the expression level of p62 was significantly increased(P<0.05).Conclusion Hcy can induce autophagy of renal podocytes.Inhibiting the expression of TRPC6 can significantly reduce the autophagy damage to podocytes.

Objective: To explore the mechanism of intestinal tissue damage induced by macrophages activated by WNT2B high-expressed fibroblasts. Methods: This study involved biological information analysis, pathological tissue research and cell experimental research. The biological information of the colon tissue from the children with inflammatory bowel disease in previous study was analyzed again with single-cell sequencing. The pathological tissues were collected by colonoscopy from 10 children with Crohn's disease treated in the Department of Gastroenterology of Guangzhou Women and Children's Medical Center from July 2022 to September 2022. According to the findings of colonoscopy, tissues with obvious inflammation or ulceration were classified as the inflammatory group, while tissues with slight inflammation and no ulceration were classified as the non-inflammatory group. HE staining was performed to observe the pathological changes of the colon tissues. Macrophage infiltration and CXCL12 expression were detected by immunofluorescence. In terms of cell experiments, fibroblasts transfected with WNT2B plasmid or empty plasmid were co-cultured with salinomycin treated or non-treated macrophages, respectively; the expression of proteins through Wnt classical pathway were detected by western blotting. Macrophages treated with SKL2001 were used as the experimental group, and those with phosphate buffer as the control group. The expression and secretion of CXCL12 in macrophages were detected by quantitative Real-time PCR and enzyme-linked immunosorbent assay (ELISA). T-test or rank sum test were used for the comparison between groups. Results: Single-cell sequencing analysis suggested that macrophages were the main cells in inflammatory bowel disease colon tissue, and there was interaction between WNT2B high-expressed fibroblasts and macrophages. HE staining of the 10 patients ((9.3±3.8) years old, 7 males and 3 females) showed that the pathological score of colon tissue in the inflammatory group was higher than that in the non-inflammatory group (4 (3, 4) vs. 2 (1, 2) points, Z=3.05, P=0.002). Tissue immunofluorescence indicated that the number of infiltrating macrophages in the inflammatory group was significantly higher than that in the non-inflammatory group under high power field of view (72.8±10.4 vs.8.4±3.5, t=25.10, P<0.001), as well as the number of cells expressing CXCL12 (14.0±3.5 vs. 4.7±1.9, t=14.68, P<0.001). In cell experiments, western blotting suggested an elevated level of glycogen synthase kinase-3β phosphorylation in macrophages co-cultured with fibroblast transfected with WNT2B plasmid, and salinmycin could reverse this change. Real-time PCR suggested that the transcription level of CXCL12 in the experimental group was higher than that in the control group (6.42±0.04 vs. 1.00±0.03, t=183.00, P<0.001), as well as the expression and secretion of CXCL12 by ELISA ((465±34) vs. (77±9) ng/L, t=13.21, P=0.006). Conclusion: WNT2B high-expressed fibroblasts can secrete WNT2B protein and activate the Wnt classical signaling pathway thus enhancing the expression and secretion of CXCL12 in macrophages, inducing the development of intestinal inflammation of Crohn's disease.
Child ; Male ; Humans ; Female ; Child, Preschool ; Adolescent ; Crohn Disease ; Inflammatory Bowel Diseases ; Colon ; Inflammation ; Colonoscopy ; Glycoproteins ; Wnt Proteins

Humans ; Bayes Theorem ; Age Factors ; Age Distribution ; Mouth Neoplasms/epidemiology* ; China/epidemiology* ; Mortality

OBJECTIVE: To investigate the mechanism by which fibroblasts with high WNT2b expression causes intestinal mucosa barrier disruption and promote the progression of inflammatory bowel disease (IBD). METHODS: Caco-2 cells were treated with 20% fibroblast conditioned medium or co-cultured with fibroblasts highly expressing WNT2b, with the cells without treatment with the conditioned medium and cells co-cultured with wild-type fibroblasts as the control groups. The changes in barrier permeability of Caco-2 cells were assessed by measuring transmembrane resistance and Lucifer Yellow permeability. In Caco-2 cells co-cultured with WNT2b-overexpressing or control intestinal fibroblasts, nuclear entry of β-catenin was detected with immunofluorescence assay, and the expressions of tight junction proteins ZO-1 and E-cadherin were detected with Western blotting. In a C57 mouse model of dextran sulfate sodium (DSS)-induced IBD-like enteritis, the therapeutic effect of intraperitoneal injection of salinomycin (5 mg/kg, an inhibitor of WNT/β-catenin signaling pathway) was evaluated by observing the changes in intestinal inflammation and detecting the expressions of tight junction proteins. RESULTS: In the coculture system, WNT2b overexpression in the fibroblasts significantly promoted nuclear entry of β-catenin (P < 0.01) and decreased the expressions of tight junction proteins in Caco-2 cells; knockdown of FZD4 expression in Caco-2 cells obviously reversed this effect. In DSS-treated mice, salinomycin treatment significantly reduced intestinal inflammation and increased the expressions of tight junction proteins in the intestinal mucosa. CONCLUSION Intestinal fibroblasts overexpressing WNT2b causes impairment of intestinal mucosal barrier function and can be a potential target for treatment of IBD.
Humans ; Mice ; Animals ; Caco-2 Cells ; beta Catenin/metabolism* ; Culture Media, Conditioned/pharmacology* ; Tight Junctions/metabolism* ; Intestinal Mucosa ; Inflammatory Bowel Diseases ; Tight Junction Proteins/metabolism* ; Inflammation/metabolism* ; Fibroblasts/metabolism* ; Mice, Inbred C57BL ; Glycoproteins/metabolism* ; Wnt Proteins/pharmacology* ; Frizzled Receptors/metabolism*

Objective:To investigate the effects of different analgesic methods of hydromorphone on analgesic efficacy and sleep quality in patients with refractory cancer pain.Methods:Sixty patients with refractory cancer pain who received three-step analgesic treatment in Quzhou People's Hospital from August 2018 to December 2019 and acquired poor analgesic effects were included in this study. They were randomly assigned to undergo either an intravenous patient-controlled analgesia with hydromorphone (HV group, n = 30) or an intrathecal patient-controlled analgesia with hydromorphone (HI group, n = 30) for 10 consecutive days. The analgesic efficacy in each group was evaluated using the numerical rating scale (NRS) before and 2, 4, 24, 48 hours, and 10 days after administration. The frequency of breakthrough pain (BTP) at each time point was recorded. The sedation effect of medication was evaluated using the Ramsay score. The sleep quality of patients was evaluated using the Pittsburgh sleep quality index (PSQI). The activities of CD 3+, CD 4+, and CD 4+/CD 8+ lymphocyte subsets were measured by flow cytometry at different time points. The adverse reactions within 10 days after treatment were observed and recorded. Results:Before and at each time point after treatment, there were no significant differences in NRS score, the frequency of BTP, Ramsay score, and PSQI score between the two groups (NRS score: t = 0.45, 0.91, 0.52, 1.19, 0.97, 1.92, all P > 0.05; frequency of BTP: t = 0.34, 1.88, 0.86, 1.71, 1.22, 0.76, all P > 0.05; Ramsay score: t = 0.56, 0.46, 0.63, 0.22, 0.99, 0.14, all P > 0.05; PSQI: t = 0.86, 1.25, 1.46, 1.05, 0.57, 1.93, all P > 0.05). At each time point after treatment, the activities of CD 3+, CD 4+, and CD 4+/CD 8+ cells increased in each group, and the activities of CD 3+, CD 4+, and CD 4+/CD 8+ cells in the HI group were significantly higher than those in the HV group (CD 3+: t = 3.72, 3.12, 2.85, 3.13, 2.44, all P < 0.05; CD 4+: t = 3.62, 2.45, 3.31, 3.19, 2.70; all P > 0.05; CD 4+/CD 8+: t = 3.10, 2.74, 2.83, 3.24, 3.41, all P < 0.05). The total incidence of adverse reactions was slightly, but not significantly, lower in the HI group than the HV group [14.00% (7/30) vs. 26.00% (13/30), χ2 = 2.70, P = 0.100]. Conclusion:Compared with intravenous administration of hydromorphone, intrathecal administration of hydromorphone can better effectively relieve pain, decrease the frequency of BTP, improve sleep quality, has a good sedative effect, improve immune function, and has fewer adverse reactions.