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 investigate the application effect of discharge preparation service based on theory of goal attainment on patients with cervical spinal cord injury.Methods:A retrospective cohort study was conducted to analyze the clinic data of 60 patients with cervical spinal cord injury admitted to Zhengzhou Orthopedics Hospital from January 2017 to December 2022, including 49 males and 11 females, aged 23-79 years [(52.2±13.5)years]. Patients were all treated with cervical decompression fusion and internal fixation. Patients admitted from January 2017 to December 2019 were treated with conventional nursing intervention (conventional nursing group, n=30) and patients admitted from January 2020 to December 2022 were treated with discharge preparation service based on theory of goal attainment (discharge preparation service group, n=30). The readiness for hospital discharge of the two groups was compared using the Chinese version of Readiness for Hospital Discharge Scale (RHDS) at 4 hours before discharge. The degree of cervical spinal cord dysfunction of the two groups were compared using Japanese Orthopedic Association (JOA) score before intervention, at discharge and at 6 months after discharge. The complication and unplanned readmission rates of the two groups were compared at 6 months after discharge. Results:All the patients were followed up for 6 months. At 4 hours before discharge, the scores of the three parameters of RHDS containing personal status, adaptability and anticipatory support and the total score of the discharge preparation service group were (20.9±3.5)points, (35.9±2.2)points, (30.4±3.0)points and (87.1±7.8)points respectively, higher than those of the conventional nursing group [(16.2±1.7)points, (32.5±2.2)points, (26.3±2.1)points and (75.0±5.6)points respectively] ( P<0.01). There was no statistically significant difference in the JOA score of the two groups before intervention ( P>0.05). The JOA scores of the discharge preparation service group at discharge and at 6 months after discharge were (11.8±1.7)points and (13.8±1.5)points respectively, higher than those of the conventional nursing group [(10.3±1.8)points and (11.6±1.9)points respectively] ( P<0.01). At 6 months after discharge, the complication rate of the discharge preparation service group was 6.7% (2/30), lower than that of the conventional nursing group [36.7% (11/30)] ( P<0.05). The unplanned readmission rate of the discharge preparation service group was 3.3% (1/30), lower than that of the conventional nursing group [23.3% (7/30)] ( P<0.05). Conclusion:For patients with cervical spinal cord injury, discharge preparation service based on theory of goal attainment can improve the discharge readiness, promote spinal functional recovery and reduce the complication and unplanned readmission rates.

Objective To analyze the expression level of YEATS2 in hepatocellular carcinoma(HCC)about its clinical prognosis and therapeutic value based on biological information from the cancer genome atlas(TCGA)and human protein atlas(HPA)databases.Methods The mRNA expression data and clinical information of HCC were downloaded from the TCGA database,the expression of YEATS2 between HCC tissues and normal tissues was analyzed by using the R software,and the protein expression differences were preliminary verified by the HPA database.The expression differences of YEATS2 between various clinical features of HCC were compared,and their effects on the survival of HCC patients by Kaplan-Meier method and COX regression analysis were then evaluated.Receiver operating characteristic(ROC)curves were plotted to evaluate their diagnostic values.The biological functions of YEATS2 in HCC were analyzed using gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis.The relationship between YEATS2 expression and tumor microenvironment(TME)was analyzed by the"ESTIMATE"algorithm,and its relationship with tumor-infiltrating immune cells(TIICs)was assessed by CIBERSORT.Analysis of YEATS2 expression levels to immune checkpoints and drug sensitivity was performed using the R package.Results The expression of YEATS2 was increased in HCC tissues(P=4.96e-21),and its expression level was correlated with age,clinical stage,pathological grade and T stage(all P＜0.05).Overall survival(OS)(P＜0.001)and progression-free survival(FPS)(P=0.016)were decreased in HCC patients with high expression of YEATS2,COX regression results showed that the expression level ofYEATS2 was associated with poor prognosis in HCC patients(OS:HR=2.167,95％CI:1.441～3.261,P=2.06e-04),and it was an independent risk factor for predicting poor prognosis in HCC patients(OS:HR=1.891,95％CI:1.243～2.877,P=0.003).The ROC curve suggested the AUCs for 1,3 and 5 years were 0.677,0.622 and 0.612,respectively,indicating good predictive ability.The TCGA database screened a total of 6 764 differential genes in the YEATS2 high and low expression groups,of which 4 094 genes were up-regulated and 2 670 genes were down-regulated in the YEATS2 high expression group.The results of GO and KEGG enrichment analyses showed that the differentially differentiated genes in the YEATS2 high expression group were mainly enriched in immunoregulation,and cell cycle regulation drug resistance pathway.The results of the TME score showed that the YEATS2 high expression group caused a decrease in immunity score(P＜0.01).The correlation between YEATS2 and TIICs showed that YEATS2 expression was positively correlated with the level of M0-type macrophage infiltration levels(r=0.48,P＜0.001)and 23 immune checkpoint genes(r=0.20～0.46,all P＜0.05),and was negatively correlated with the CD8+T-cells,plasma cells and monocyte(r=-0.26,-0.29,-0.30,P=0.021,0.011,0.008).Drug sensitivity analysis showed that the half maximal inhibitory concentration(IC50)of cabozantinib,lincitinib,doxorubicin,and cyclobenzaprine in patients with high expression of YEATS2 was higher than those in patients with low expression(all P＜0.01).Conclusion YEATS2 was highly expressed in HCC,and the expression level was associated with poor prognosis in HCC patients.YEATS2 can be used as a biomarker for the clinical early diagnosis,prognosis and immunotherapy of HCC,which may provide new ideas for clinical diagnosis and treatment.

In the progression of end-stage liver disease,the incidence of sleep disorders in patients with liver cirrhosis is high.Currently,western medicine treatment has obvious liver and kidney damage and adverse reactions after discontinuation,which affects the therapeutic effect.Cirrhosis and sleep disorders can be separately attributed to the category of"abdominal mass"and"insomnia"in traditional Chinese medicine.The"abdominal mass"is caused by the disorder of liver and spleen qi movement,as well as the obstruction of phlegm-dampness and blood stasis.In the development of"abdominal mass",the liver failed in ensuring free movement of qi,the spleen failed in transportation and transformation,liver yin and liver blood became insufficiency and the imbalance of yin and yang in the zang-fu organs became imbalanced,and then the ethereal soul depart from the housing,which leads to"insomnia"as an outward manifestation.Professor ZHOU Xiao-Zhou focuses on the concept of qi and blood,and points out that the pathogenesis of sleep disorder in cirrhosis is characterized by liver stagnation and spleen deficiency.He proposed that the treatment principle is to regulate the liver and spleen simultaneously,as well as to nourish the heart and calm the mind.Professor ZHOU has developed Ganyinghua Anshen Formula for treating sleep disorders in cirrhosis,which is derived from the modification of Xiangsha Liujunzi Decoction,and is composed of 14 Chinese medicines of vinegar-prepared Cyperi Rhizoma,Amomi Fructus,Coicis Semen,Dioscoreae Rhizoma,bran-fried Atractylodis Macrocephalae Rhizoma,Galli Gigerii Endothelium Corneum,Gardeniae Fructus,Codonopsis Radix,Salviae Miltiorrhizae Radix et Rhizoma,Citri Reticulatae Pericarpium,Sclerotium Poriae Pararadicis,Longan Arillus,Albiziae Cortex,and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle.The formula has achieved remarkable clinical effect.The clinical experience of Professor ZHOU Xiao-Zhou for treating sleep disorders in cirrhosis through regulating the liver and spleen simultaneously will provide a reference for its clinical treatment with traditional Chinese medicine.

Objective To develop a matrix metalloproteinase(MMP)-responsive hyaluronic acid(HA)-based controlled-release material for brain-derived neurotrophic factor(BDNF)to provide a novel therapeutic strategy for intervention and repair of traumatic brain injury(TBI).Methods HA was modified with amination,followed by condensation with Suflo-SMCC carboxyl group to form amide,and then linked with glutathione(GSH)to synthesize HA-GSH.The recombinant glutathione S-transferase(GST)-tissue inhibitor of metalloproteinase(TIMP)-BDNF(GST-TIMP-BDNF)expression plasmid was constructed using molecular cloning technique with double enzyme digestion by Bam H Ⅰ and Eco R Ⅰ.The recombinant GST-TIMP-BDNF protein was expressed in the Escherichia coli prokaryotic expression system,and purified by ion exchange chromatography,confirmed by Western blotting.MMP diluents were supplemented with PBS,MMP inhibitor marimastat,and varing concentrations(0.4,0.6,0.8 mg/ml)of GST-TIMP-BDNF or GST-BDNF.MMP-2 activity was analyzed using an MMP activity detection kit to evaluate the inhibitory effect of the recombinant protein on MMP.Primary rat neurons were extracted and cultured to establish an iron death model induced by RSL3.The effect of recombinant protein GST-TIMP-BDNF on neuronal injury was detected by immunofluorescence staining.Results MRI hydrogen spectrum identification confirmed the successful synthesis of HA-GSH.Western blotting results showed the successful expression of the recombinant protein GST-TIMP-BDNF containing the GST tag using the E.coli prokaryotic expression system.MMP activity detection results indicated that the recombinant protein GST-TIMP-BDNF had a superior inhibitory effect on MMP-2 activity compared to GST-BDNF(P<0.05).Immunofluorescence staining results showed a significant increase in fluorescence intensity in rat neurons treated with GST-TIMP-BDNF after RSL3 induction(P<0.05).Conclusion A MMP-responsive HA-based BDNF controlled-release material has been successfully developed,exhibiting a protective effect on neuron damage.

Aim To investigate the effect of vanillin on the regulation of cyclic guanylate adenylate synthetase(cGAS)/stimulator of interferon gene(STING)signa-ling pathway on hepatic tissue injury in rats with intra-hepatic cholestasis(IC).Methods SD rats were randomly divided into normal group,IC group,vanillin group,cGAS overexpression group,and vanillin+cGAS overexpression group,with continuous adminis-tration for seven days.The body weight,liver weight and liver to body weight ratio of rats were measured.Liver function(ALT,AST,ALP,LDH),IC(TBIL,TBA)and liver fibrosis(HA,LN,PC Ⅲ)index were determined by ELISA.Liver pathology and fibrosis were observed using HE and Masson staining,and col-lagen volume fraction was calculated.The expression of cGAS/STING pathway related proteins in liver tissue was detected by Western blot.Results Vanillin could improve liver pathology and fibrosis,increase body weight,and decrease liver weight,ALT,AST,ALP,LDH,TBIL,TBA,HA,LN,PC Ⅲ,collagen volume fraction,cGAS,STING protein in IC rats(P＜0.05).Overexpression of cGAS could reverse the effects of vanillin on the above indicators in IC rats(P＜0.05).Conclusions Vanillin may improve liver function,IC,liver fibrosis,and liver tissue damage in IC rats by downregulating the cGAS/STING signaling pathway.

Objective To predict the potential distribution areas of three kinds of Stemonae Radix in China;To provide reference for the cultivation site selection of three kinds of Stemonae Radix;To reduce the confusion of Stemonae Radix medicinal materials in the selection of planting areas.Methods Totally 130 pieces of geographical distribution information of Stemona tuberosa Lour.,52 pieces of geographical distribution information of Stemona japonica(Bl.)Miq.,and 52 pieces of geographical distribution information of Stemona sessilifolia(Miq.)Miq.were combined with 91 environmental variables and 3 topographic factors.The maximum entropy(MaxEnt)model and geographic information system software ArcGIS 10.2 were used to predict the potential distribution areas of the three kinds of Stemonae Radix.Results Stemona tuberosa Lour.is mainly distributed in the southern provinces,including eastern Sichuan,western Chongqing,eastern Guangdong,southwestern Fujian,Taiwan,Hainan,southern Yunnan and the junction of Guizhou,Guangxi and Yunnan provinces.The high suitable area is about 293 983 km2,and the medium suitable area is about 519 667 km2.Stemona japonica(Bl.)Miq.is mainly distributed in the plain area of the middle and lower reaches of the Yangtze River,including northwestern and eastern Zhejiang,northern Jiangxi,eastern Hubei,southern Anhui,southern Jiangsu and northern Fujian.The high suitable area is about 140 320 km2,and the medium suitable area is about 188 752 km2.Stemona sessilifolia(Miq.)Miq.is mainly distributed in the North China Plain,including northeastern Hubei,southern Henan,central Anhui,western Jiangsu and central Shandong.The high suitable area is about 198 568 km2,and the medium suitable area is about 198 626 km2.Conclusion The results have important guiding significance for the standardized cultivation of three kinds of Stemonae Radix,and also provide reference for the cultivation of Stemonae Radix.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

In this study,the immunological characteristics of the PhoP protein were explored with a two-component system of Mycobacterium tuberculosis(Mtb).Bioinformatics was used to predict the B and T cell epitopes of the PhoP protein.A re-combinant expression plasmid was constructed by PCR analysis of the phoP sequence and cloning into the prokaryotic expres-sion vector pET-28a(+).Competent Escherichia coli BL21 cells were transformed with the recombinant plasmid and expres-sion was induced with IPTG.The recombinant PhoP protein was purified by affinity chromatography.Serum levels of PhoP-specific antibodies in Mtb-infected mice and tuberculosis(TB)patients were analyzed with an ELISA.BALB/c mice were im-munized with the PhoP recombinant protein by intramuscular injection.Sera of mice were collected and antibody titers were detected with an ELISA and specificity was assessed by West-ern blot analysis.Mouse splenocytes were isolated and the pro-portions of IFN-y-positive cells and cytokine levels were detec-ted with an ELISpot and ELISA,respectively.Bioinformatics i-dentified 24 B cell and 11 T cell epitopes of the PhoP protein.A prokaryotic recombinant vector of PhoP was successfully con-structed and the recombinant PhoP protein was obtained by purification.Specific antibody levels to PhoP in sera of Mtb in-fected mice and TB patients increased significantly,with preci-sion of 99.9％and 82.5％,and specificity of 100％,respectively.PhoP protein immunization successfully induced production of specific antibodies in mice.Stimulated by antigens in vitro,IL-2 and IFN-γ levels were significantly increased in the splenocytes of immunized mice.Immunization with the PhoP protein induce a humoral immune response and Thl-dominated cellular immu-nity,indicating that the PhoP protein was immunogenic with diagnostic efficacy for TB.These results lay a foundation to clari-fy the role of PhoP in Mtb infection and application for diagnosis and prevention of TB.

Objective To explore association between childhood trauma,resilience and non-suicidal self-injury in adolescents.Methods One hundred and fifty-eight first-episode adolescent patients with mood disorders were selected and divided into NSSI group(n=94)and non-NSSI group(n=64)based on presence or absence of NSSI.The Hamilton depression scale(HAMD),Hamilton anxiety scale(HAMA),childhood trauma questionnaire(CTQ-SF)and Connor-Davidson resilience scale(CD-RISC)were used to evaluate the depression and anxiety symptoms,childhood trauma and resilience.Results There were more cases of younger age(16.17±1.67 vs.16.73±1.37),lower education level(30.9% vs.15.6% ),left behind experience(48.9% vs.29.7% ),school bullying(46.8% vs.25.0% ),suicide ideation(85.1% vs.37.5% )and history of attempted suicide(29.8% vs.6.3% )in the group with NSSI compared to those without NSSI.The HAMD score(27.99±5.94 vs.24.19±5.19),HAMA score(18.02±5.94 vs.15.45±4.99),CTQ total score(48.43±15.40 vs.41.97±9.75),emotional abuse score(12.77±6.06 vs.10.19±4.06),and emotional neglect score(11.40±5.34 vs.9.14±3.55)were higher in the group with NSSI,and the differences between the two groups were significant(P<0.05).The total scores of psychological resilience(39.83±10.27 vs.28.66±12.75),resilience(19.59±4.92 vs.12.28±6.47),strength(12.03±3.98 vs.9.99±4.67),and optimism(8.98±2.97 vs.6.47±3.73)in the group without NSSI were higher than those in the group with NSSI(P<0.05).Logistic regression analysis showed that left behind experience(OR=4.494,95% CI:1.192-16.940),school bullying(OR=5.983,95% CI:1.329-26.945),suicidal ideation(OR=13.225,95% CI:2.908-60.146),history of attempted suicide(OR=16.769,95% CI:1.845-152.379),HAMD(OR=1.264,95% CI:1.046-1.626),emotional abuse(OR=1.327,95% CI:1.093-1.612),and resilience(OR=0.468,95% CI:0.266-0.823)were significantly associated with adolescent mood disorders with NSSI(P<0.05).Conclusion Left behind experience,campus bullying,suicidal ideation and attempted suicide,emotional abuse,degree of depression,and psychological resilience may be associated with NSSI behavior in adolescents with mood disorders.