Background: and Purpose Guillain–Barré syndrome (GBS) is an autoimmune-mediated disorder characterized by demyelinating or axonal injury of the peripheral nerve. Our aim is to determine whether serum amyloid A (SAA) is a biomarker of demyelinating injury and disease severity in patients with GBS. Methods: This study retrospectively enrolled 40 patients with either the demyelinating or axonal GBS and sex- and age-matched controls with other neurological diseases as well as healthy subjects. The demographic and clinical features at entry were collected. The serum levels of the SAA isoforms SAA1, SAA2, and SAA4 were determined in the patients with GBS and the controls using the enzyme-linked immunosorbent assay and analyzed for the associations between levels of different SAA isoforms and the clinical features of the patients. Results: The levels of SAA2 and SAA4 were significantly higher in patients with GBS than in both the other neurological disease controls and the healthy subjects (p<0.05 for all). The level of SAA1 did not differ between patients with GBS and the controls. The level of SAA2 was considerably higher in GBS patients with antecedent infection than in those without infection (p=0.020). The levels of different SAA isoforms were not associated with the disease severity or other clinical features of patients with GBS (p>0.05 for all). Conclusions Increased levels of SAA2 and SAA4 may only represent the acute inflammatory status and so cannot be utilized as biomarkers of the disease severity or demyelinating injury in patients with GBS.

Background: and Purpose Guillain–Barré syndrome (GBS) is an autoimmune-mediated disorder characterized by demyelinating or axonal injury of the peripheral nerve. Our aim is to determine whether serum amyloid A (SAA) is a biomarker of demyelinating injury and disease severity in patients with GBS. Methods: This study retrospectively enrolled 40 patients with either the demyelinating or axonal GBS and sex- and age-matched controls with other neurological diseases as well as healthy subjects. The demographic and clinical features at entry were collected. The serum levels of the SAA isoforms SAA1, SAA2, and SAA4 were determined in the patients with GBS and the controls using the enzyme-linked immunosorbent assay and analyzed for the associations between levels of different SAA isoforms and the clinical features of the patients. Results: The levels of SAA2 and SAA4 were significantly higher in patients with GBS than in both the other neurological disease controls and the healthy subjects (p<0.05 for all). The level of SAA1 did not differ between patients with GBS and the controls. The level of SAA2 was considerably higher in GBS patients with antecedent infection than in those without infection (p=0.020). The levels of different SAA isoforms were not associated with the disease severity or other clinical features of patients with GBS (p>0.05 for all). Conclusions Increased levels of SAA2 and SAA4 may only represent the acute inflammatory status and so cannot be utilized as biomarkers of the disease severity or demyelinating injury in patients with GBS.

Background: and Purpose Guillain–Barré syndrome (GBS) is an autoimmune-mediated disorder characterized by demyelinating or axonal injury of the peripheral nerve. Our aim is to determine whether serum amyloid A (SAA) is a biomarker of demyelinating injury and disease severity in patients with GBS. Methods: This study retrospectively enrolled 40 patients with either the demyelinating or axonal GBS and sex- and age-matched controls with other neurological diseases as well as healthy subjects. The demographic and clinical features at entry were collected. The serum levels of the SAA isoforms SAA1, SAA2, and SAA4 were determined in the patients with GBS and the controls using the enzyme-linked immunosorbent assay and analyzed for the associations between levels of different SAA isoforms and the clinical features of the patients. Results: The levels of SAA2 and SAA4 were significantly higher in patients with GBS than in both the other neurological disease controls and the healthy subjects (p<0.05 for all). The level of SAA1 did not differ between patients with GBS and the controls. The level of SAA2 was considerably higher in GBS patients with antecedent infection than in those without infection (p=0.020). The levels of different SAA isoforms were not associated with the disease severity or other clinical features of patients with GBS (p>0.05 for all). Conclusions Increased levels of SAA2 and SAA4 may only represent the acute inflammatory status and so cannot be utilized as biomarkers of the disease severity or demyelinating injury in patients with GBS.

Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

Synthetic biology is an emerging interdisciplinary field at the convergence of biology, engineering, and computer science. It employs a bottom-up approach to progressively design biological parts, devices, and circuits, aiming to create artificial biological systems not found in nature or to redesign existing biological systems for specific purposes. With the rapid development of the synthetic biology industry, there is an increasing demand for large complex genetic circuits. However, the traditional trial-and-error methods, heavily reliant on empirical knowledge, have limited efficiency and success rates of parts/circuits construction, thereby impeding the innovation and technology translation for synthetic biology. These limitations have prompted a paradigm shift from labor-intensive, experience-driven trial-and-error models towards standardized, intelligent engineering approaches. Machine learning, capable of uncovering hidden structures and relationships within biological data, offers robust support for the intelligent design of synthetic biological parts and genetic circuits. Here, we review commonly used machine learning algorithms and analyze their typical applications in designing biological parts (e.g., synthetic promoters, RNA regulatory elements, and transcription factors) and simple genetic circuits. Additionally, we discuss the primary challenges in machine learning-aided design and propose potential solutions. Lastly, we envision the future trend of integrating machine learning with synthetic biological system design, highlighting the importance of interdisciplinary collaboration.
Synthetic Biology/methods* ; Machine Learning ; Gene Regulatory Networks ; Algorithms

Objective:To explore the distribution and characteristics of microbiota in the lower reproductive tract of patients with cervical squamous cell carcinoma infected by human papilloma virus (HPV) 16 subtype.Methods:A prospective, cross-sectional study was conducted. A total of 6 patients with HPV16 single subtype positive cervical squamous cell carcinoma admitted to the Cancer Hospital of Chinese Academy of Medical Sciences from August 2019 to June 2020 were selected as cervical carcinoma group, and 6 healthy women who did not indicate abnormalities in thin-based layer cytology test (TCT) during the same period among the physical examination population and had HPV negative test result were selected as the healthy control group. A sterile cotton swab was used to collect secretions from the posterior cervical fornix in patients before antitumor treatment and healthy controls during physical examination. The high variable region of the 16S rRNA gene V1-V2 of the bacteria was amplified by using next generation sequencing (NGS), and then the distribution and characteristics of the bacteria were analyzed.Results:The age of cervical cancer group and the healthy control group was (51±8) years and (48±3) years, respectively, and the difference in age between the both groups was statistically significant ( t= 0.63, P= 0.540). The patients of both groups had reproductive history and no smoking experience. Alpha diversity analysis showed that compared with the healthy control group, the sobs ( t= 3.25, P= 0.009) and chao ( t= 2.91, P= 0.016) indexes were higher in cervical cancer group, and the differences were statistically significant. The shannon index was higher ( t= 2.07, P= 0.065) and simpson index was lower ( t= 1.74, P= 0.113) in cervical cancer group, while the difference was not statistically different. Data dimensionality reduction analysis in principal coordinate analysis (PCoA) based on bray-curtis distance showed that the difference in Beta diversity between the healthy control group and cervical cancer group was statistically significant ( R2= 0.154, P = 0.018). At the phylum level, the proportion of Firmicutes in cervical cancer group was lower than that in the healthy control group (30.21% vs. 68.28%), while the proportion of Bacteroidetes in cervical cancer group decreased slightly (6.87% vs. 8.11%); and the proportion of Actinobacteria (26.91% vs. 14.42%) and Proteobacteria (27.33% vs. 0.67%) had an increase in cervical cancer group. At the genus level, compared with the healthy control group, the proportion of Lactobacillus and Corynebacterium decreased in cervical cancer group, and loss of dominant flora could be detected; while Rhodococcus, Klebsiella and Aerococcus increased significantly in cervical cancer group. The bacteria species in cervical cancer group was increased compared with the healthy control group. Linear discriminant analysis (LDA) showed that Rhodococcus (LDA = 5.04), Klebsiella (LDA = 4.71), Enterobacter (LDA = 4.29), Ralstonia (LDA = 4.28), Ochrobactrum (LDA = 4.23) and Veillonella (LDA = 4.14) were the distinctive microbiota of cervical cancer group at the genus level. At the phylum level, Firmicutes (LDA = 5.23) in the healthy control group could be considered as a marker species. At the species level, the proportions of Rhodococcus ( P = 0.025), Ralstonia ( P = 0.045), Veillonella ( P = 0.044), Paraburkholderia ( P = 0.045), Pseudomonas ( P = 0.043) in cervical cancer group were increased compared with the healthy control group, and the differences were statistically significant. Conclusions:HPV16 single positive patients with cervical squamous cell carcinoma show the characteristics such as the increased diversity and richness of the lower reproductive tract microbiota compared with the healthy controls, while the abundance of Lactobacillus decreases. Rhodococcus and Klebsiella could serve as symbolic microbial in the lower reproductive tract. However, further studies still need to be verified.

Objective To explore the mechanism of butylphthalide(NBP)in regulating microglia acti-vation and inflammatory cytokine expression in the hippocampus of the mouse model of delayed encephalopathy af-ter carbon monoxide poisoning(DEACMP).Methods Wild-type C57 adult mice with normal cognitive function were selected,and DEACMP was modeled by static inhalation of carbon monoxide.The mice were randomized in-to three groups:DEACMP,control,and NBP.The NBP group was administrated with NBP suspension at 6 mg/kg by gavage for 21 days,and the DEACMP and control groups were administrated with the same amount of vegeta-ble oil by gavage.The hippocampal injury was observed by HE staining.The protein level of ionized calcium-bind-ing adapter molecule 1(IBA1)was determined by Western blotting,and the levels of downstream inflammatory cytokines were measured by ELISA.Results Compared with the control group,the DEACMP and NBP groups showed prolonged escape latency(P=0.001,P=0.029),reduced nerve cells(P=0.001,P=0.035),up-regulated expression of IBA1(P=0.001,P=0.042),increased mean fluorescence intensity of IBA1(P=0.001,P=0.021),and elevated levels of tumor necrosis factor-α(TNF-α)(P=0.002,P=0.024),inter-leukin(IL)-6(P=0.001,P=0.015),and IL-1β(P=0.001,P=0.023).Compared with the DEACMP group,the NBP group showed shortened escape latency(P=0.025),increased nerve cells(P=0.039),down-regulated expression of IBA1(P=0.035),decreased average fluorescence intensity of IBA1(P=0.031),and lowered levels of TNF-α(P=0.028),IL-6(P=0.037),and IL-1 β(P=0.034).Conclusion NBP can inhibit the activation of microglia and reduce the expression of inflammatory factors,thereby alleviating cog-nitive dysfunction and brain tissue damage caused by DEACMP.

Objective:To propose a novel refined subtyping of Neer type Ⅵ proximal humerus fracture-dislocation and explore its clinical application.Methods:A retrospective study was conducted to analyze the data of 36 patients who had been admitted to Department of Orthopaedics, Beijing Friendship Hospital between January 2018 and December 2022 for surgical treatment with proximal humeral internal locking system (PHILOS) for Neer type Ⅵ proximal humerus fracture-dislocation. There were 25 males and 11 females with an age of (46.1±4.7) years. According to the fracture-dislocation and the separation between the humeral head and the stem, the patients with Neer type Ⅵ proximal humerus fracture-dislocation were further subdivided into 3 subtype groups (known as STAB subtypes): subtype-T group (dislocation of the shoulder joint with macro-capitellar fracture, n=14), subtype-A group (proximal humerus fracture-dislocation without separation of the humeral head from the humeral stem, n=12), and subtype-B group (dislocation of the proximal humerus fracture with separation of the humeral head from the humeral stem, n=10). STAB subtyping was performed on the same imaging data from all the patients at admission and 2 weeks later by 4 surgeons with different qualifications. Interobserver and intraobserver agreements of the STAB typing were verified. The operation time, fracture healing time, visual analogue scale (VAS) pain score, Constant-Murley score, and complications were recorded for patients in the 3 subtype groups. Results:The differences in the preoperative general data were not statistically significant between the 3 subtype groups, indicating comparability ( P>0.05). All patients were followed up for (11.2±4.2) months. The inter-observer and intra-observer Kappa values for STAB subtyping were 0.94 and 0.95, respectively. For subtype-T group, subtype-A group, and subtype-B group, respectively, the operation time was (68.9±5.6) min, (90.0±5.2) min, and (113.0±9.2) min; the fracture healing time was (9.0±0.8) weeks, (10.3±1.2) weeks, and (11.8±0.9) weeks; the VAS scores at the last follow-up were 1.0(1.0, 2.0) points, 2.0(1.0, 2.0) points, 2.0(2.0, 3.0) points; the Constant-Murley scores at the last follow-up were (83.6±2.8) points, (74.5±3.0) points, and (62.7±5.5) points. The differences between the 3 subtype groups in the above items were statistically significant ( P<0.05). The overall success rate of closed reduction was 61.1% (22/36). In subtype-T, subtype-A, and subtype-B groups, respectively, the number of patients with successful closed reduction was 13, 7, and 2, while complications occurred in 2, 3, and 6 patients. The differences in closed reduction and complications among the 3 groups were statistically significant ( P<0.05). Conclusions:The STAB subtyping proposed in this study demonstrates strong intra- and inter-group consistency. Because the refined STAB subtyping can reveal differences among all the Neer type Ⅵ proximal humeral fractures and dislocations, it may provide more precise guidance for personalized clinical decision-making.

Objective To preliminarily investigate the effects of estradiol on retinal microglia and retinal ganglion cells(RGCs)in rats with glucocorticoid-induced ocular hypertension(OHT).Methods Thirty-six male SD rats(36 eyes)were randomly divided into a control group,an OHT group,and an OHT estradiol-treated group(E2-OHT group),with 12 rats in each group.Among them,the rats in the OHT group and the E2-OHT group were given dexamethasone sodi-um phosphate injection under the conjunctiva,and the rats in the control group were injected with the same volume of ster-ile normal saline.Two weeks after modeling,the rats in the E2-OHT group were treated with estradiol eye drops in addition to subconjunctival injection of dexamethasone sodium phosphate.The eyeballs of all rats were removed 4 weeks after mod-eling.The changes in the number of RGCs and the activation of microglia were observed after immunofluorescence stai-ning,the expression levels of Brn3a and Iba1 proteins in the retina were detected by Western blot,and the relative expres-sion levels of tumor necrosis factor α(TNF-α)and interleukin 1 β(IL-1 β)mRNA were detected by real-time quantitative polymerase chain reaction.Results Among the three groups,the intraocular pressure(IOP)of rats showed no signifi-cant difference before modeling(all P＞0.05),but showed a significant difference at 1 week,2 weeks,3 weeks and 4 weeks after modeling(all P＜0.01).Compared with the control group,the IOP of rats in the OHT group at 1 week,2 weeks,3 weeks and 4 weeks after modeling increased significantly(all P＜0.01).Compared with the OHT group,the IOP of rats in the E2-OHT group showed no significant difference at 1 week and 2 weeks after modeling(both P＞0.05),but decreased significantly at 3 weeks and 4 weeks after modeling(both P＜0.01).The immunofluorescence staining results showed that the retinal microglia of rats in the control group were mainly concentrated in the inner plexiform layer,while the retinal microglia of rats in the OHT group migrated to the ganglion cell layer and had morphological changes(amoebic activation state).The morphology and distribution of rat retinal microglia in the E2-OHT group were basically the same as the retinal staining results of rats in the control group.Compared with the control group,the number of RGCs in the OHT group decreased,the relative expression levels of TNF-α and IL-1β mRNA and Iba1 protein increased,while the expression level of Brn3a protein decreased,and the differences were statistically significant(all P＜0.05).Compared with the OHT group,the rats in the E2-OHT group had an increased number of RGCs,a decreased relative expression level of TNF-α and IL-1 β mRNA and Ibal protein,and an increased expression level of Brn3a protein(all P＜0.05).Conclusion Estradiol can inhibit the activation of microglia,reduce the expression of TNF-α and IL-1β in the retina of rats with OHT,and reduce the damage to RGCs.

Objective To observe the effect of neuregulin-1(NRG-1)on retinal ganglion cells(RGCs)in Zucker Di-abetic Fatty(ZDF)rats and explore the mechanism of NRG-1 in exerting neuroprotective effects on the retina.Methods Twenty-four 8-week-old male ZDF rats(24 eyes)were selected.Diabetic obese rat models were established by feeding a high-fat and high-sugar diet(Purina 5008).After 16 weeks,they were randomly divided into the ZDF group and the NRG-1 group(12 rats in each group).Rats in the NRG-1 group received intravitreal injection of human recombinant NRG-1(once a week for a total of two times),rats in the ZDF group were used as negative controls,and Zucker lean control(ZLC)rats were selected as blank controls(ZCL group).The changes in the number of RGCs in rats of each group were observed by immunofluorescence staining.The protein expressions of NOD-like receptor family pyrin domain containing protein 3(NLRP3),Caspase-1,and Gasdermin D(GSDMD)in the retina of rats in each group were observed by immuno-histochemistry and Western blot.Results After 16 weeks of eating a high-fat diet,compared with the ZLC group,the fasting blood glucose of rats in the ZDF group significantly increased(P＜0.001).The results of immunofluorescence stai-ning showed that the RGCs of rats in the ZLC group were continuously and neatly arranged;compared with the ZLC group,the number of RGCs of rats in the ZDF group significantly decreased(P＜0.001);compared with the ZDF group,the num-ber of RGCs of rats in the NRG-1 group significantly increased(P＜0.01).The immunohistochemical results showed that there were statistically significant differences in the average optical density values of NLRP3,Caspase-1 and GSDMD in the retina of rats in each group(all P＜0.01);compared with the ZLC group,the average optical density values of NLRP3,Caspase-1 and GSDMD in the retina of rats in the ZDF group were higher(all P＜0.01);compared with the ZDF group,the average optical density values of NLRP3,Caspase-1 and GSDMD in the retina of rats in the NRG-1 group significantly de-creased(all P＜0.05).Western blot results showed that there were statistically significant differences in the protein ex-pression levels of Brn3a,NLRP3,Caspase-1 and GSDMD in the retina of rats among all groups(all P＜0.01);compared with the ZLC group,the protein expression of Brn3a significantly decreased,while the protein expressions of NLRP3,Caspase-1 and GSDMD significantly increased in the ZDF group(all P＜0.01);compared with the ZDF group,the protein expression of Brn3a significantly increased,while the protein expressions of NLRP3,Caspase-1 and GSDMD significantly decreased in the NRG-1 group(all P＜0.01).Conclusion After retinal lesions occur in diabetic rats,NLRP3,Caspase-1 and GSDMD are all significantly activated.NRG-1 can reduce the expressions of NLRP3,Caspase-1 and GSDMD,reducing damage to RGCs.