AIM: To investigate the expression levels of follistatin-like protein 1(FSTL1)and a disintegrin and metalloproteinase with thrombospondin type 1 motif 13(ADAMTS13)in the serum of patients with proliferative diabetic retinopathy(PDR), and to analyze their predictive value for visual disability.METHODS:A prospective study. From January 2022 to September 2024, patients with diabetes retinopathy(DR)were selected from Fengfeng General Hospital of North China Medical and Health Group. They were divided into non-proliferative diabetes retinopathy(NPDR)group and PDR group. In the same period, simple type 2 diabetes(T2DM)patients were selected as T2DM group, and healthy people were selected as control group. Enzyme linked immunosorbent assay was used to detect serum levels of FSTL1 and ADAMTS13. Patients in the PDR group were followed up for 12 mo to observe the occurrence of visual disability, and were accordingly divided into a non-visual disability group and a visual disability group. Risk factors for visual disability in PDR patients were analyzed by Logistic regression, and the predictive value was assessed via receiver operating characteristic(ROC)curves and decision curve analysis(DCA).RESULTS: This study included 220 DR patients, including 119 cases in the NPDR group and 101 cases in the PDR group. There were 120 cases in the T2DM group and 120 cases in the control group. The control group had an age of 56.92±8.15 y, with 66 males and 54 females. The T2DM group had an age of 57.21±8.30 y, with 63 males and 57 females. The NPDR group had an age of 57.36±8.25 y, with 60 males and 59 females. The PDR group had an age of 57.45±8.22 y, with 53 males and 48 females. There were 68 patients without visual disability and 33 patients with visual disability in the PDR group. The non-visual disability group had a mean age of 57.26±8.14 y, including 35 males and 33 females. The visual disability group had a mean age of 57.85±8.37 y, including 18 males and 15 females. Diabetes duration was progressively longer in the T2DM, NPDR, and PDR groups. Levels of fasting plasma glucose(FPG), glycated hemoglobin(HbA1c), and FSTL1 progressively increased, while ADAMTS13 levels progressively decreased across the control, T2DM, NPDR, and PDR groups(all P<0.001). Within the PDR group, serum FSTL1 levels increased sequentially while ADAMTS13 levels decreased sequentially in patients with grade Ⅳ, Ⅴ and Ⅵ lesions(all P<0.001). The visual disability group had significantly longer diabetes duration and higher levels of FPG, HbA1c, and FSTL1, as well as lower ADAMTS13 levels, compared to the non-visual disability group(all P<0.001). The diabetes duration, FSTL1, ADAMTS13 were the influencing factors of visual disability in PDR patients(all P<0.001). ROC curve and DCA curve analysis showed that the predictive value of combined detection of serum FSTL1 and ADAMTS13 levels is higher than that of individual measurement(all P<0.05).CONCLUSION:PDR patients have higher serum FSTL1 levels and lower ADAMTS13 levels. Both are related to the severity of PDR. The combined measurement has a high predictive value for visual disability in PDR patients.

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

Objective:To document the effectiveness of ultrasound-guided genicular nerve block (GNB) in treating knee osteoarthritis (KOA).Methods:A total of 92 KOA patients were randomly divided into an observation group and a control group with 46 in each. Those in both groups were treated conventionally, including with non-steroidal anti-inflammatory drugs, acupuncture, ultrasound, laser irradiation and manipulation therapy. The observation group additionally underwent ultrasound-guided genicular nerve block treatment, once a week for 2 weeks. Pain scoring on a visual analog scale (VAS), the Western Ontario and McMaster University Osteoarthritis Index (WOMAC) and a 6-minute walk test (6MWT) were used to evaluate everyone before and after the treatment, and then 8 weeks later.Results:In the observation group the average VAS rating [(3.54±2.00) at week 2 and (4.13±2.04) at week 8] and the average WOMAC subscale and total scores [(36.91±16.91) at week 8] had improved significantly right after the experiment and 8 weeks later. But in the control group this was true only right after the treatment. The observation group also demonstrated superior improvements in 6MWT distance at week 2 [(434.22±125.19)m] and week 8 [(446.35±126.45)m] compared to both its own baseline and the control group.Conclusions:Ultrasound-guided genicular nerve block is a rapid, precise, effective, and long-lasting intervention for alleviating pain, improving knee function and enhancing walking endurance in KOA patients.

Objective:To document the effectiveness of ultrasound-guided genicular nerve block (GNB) in treating knee osteoarthritis (KOA).Methods:A total of 92 KOA patients were randomly divided into an observation group and a control group with 46 in each. Those in both groups were treated conventionally, including with non-steroidal anti-inflammatory drugs, acupuncture, ultrasound, laser irradiation and manipulation therapy. The observation group additionally underwent ultrasound-guided genicular nerve block treatment, once a week for 2 weeks. Pain scoring on a visual analog scale (VAS), the Western Ontario and McMaster University Osteoarthritis Index (WOMAC) and a 6-minute walk test (6MWT) were used to evaluate everyone before and after the treatment, and then 8 weeks later.Results:In the observation group the average VAS rating [(3.54±2.00) at week 2 and (4.13±2.04) at week 8] and the average WOMAC subscale and total scores [(36.91±16.91) at week 8] had improved significantly right after the experiment and 8 weeks later. But in the control group this was true only right after the treatment. The observation group also demonstrated superior improvements in 6MWT distance at week 2 [(434.22±125.19)m] and week 8 [(446.35±126.45)m] compared to both its own baseline and the control group.Conclusions:Ultrasound-guided genicular nerve block is a rapid, precise, effective, and long-lasting intervention for alleviating pain, improving knee function and enhancing walking endurance in KOA patients.

Objective To evaluate the renoprotective effects of zinc(Zn)supplementation in diabetes kidney disease(DKD)and to explore its impact on the nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.Methods A total of 12 male OVE26 mice(spontaneous type 1 diabetes mellitus mice)aged 3 months and weighing approximately 24-27 g were selected and randomly assigned to a diabetes mellitus(DM)group and a zinc-treated DM(DM/Zn)group(n=6 each).In addition,12 age-matched male FVB mice weighing approximately 27-30 g were selected and randomly assigned to a non-diabetic control(Ctrl)group and a zinc-treated(Zn)group(n=6 each).Mice in the DM/Zn and Zn groups were given zinc supplementation for 3 months,with each mouse receiving 5 mg/kg of zinc sulfate by gavage every other day.Mice in the DM and Ctrl groups were given the same volume of normal saline.At the end of the experiment,the albumin-to-creatinine ratio(ACR)in urine was used as an indicator to evaluate renal function.Sirius red staining was performed to assess renal fibrosis in each group of mice.Western blotting was performed to determine the expression of fibrotic growth factors,including connective tissue growth factor(CTGF)and transforming growth factor-β1(TGF-β1),in renal tissue,and the protein expression of Nrf2,an antioxidant substance,and the protein expression levels of its downstream targets,including NAD(P)H quinone dehydrogenase 1(NQO1),heme oxygenase 1(HO-1),superoxide dismutase(SOD)-1,SOD-2,and catalase(CAT).Results 1)Compared to the Ctrl group,the urinary protein secretion levels of mice in the DM group exhibited progressive increase.After 3 months of zinc supplementation treatment,the urinary protein secretion levels of mice in the DM/Zn group decreased Compared to that of mice in the DM group,and the difference was statistically significant(P<0.05).2)Compared to that in the Ctrl group,the collagen deposition in the renal tissues of mice in the DM group increased,and the difference was statistically significant(P<0.05),while no obvious change was observed in mice in the DM/Zn group.Compared to the Ctrl group,mice in the DM group exhibited increased expression levels of CTGF and TGF-β1 in the renal tissues,but the expression levels decreased after zinc supplementation treatment,with the differences being statistically significant(P<0.05).3)Compared to that of the Ctrl group,the expression level of Nrf2 in the renal tissues of mice in the Zn and DM groups increased,and the level of Nrf2 in the renal tissues of mice in the DM/Zn group showed a further increase,with the differences being statistically significant(P<0.05).4)Compared to those of the Ctrl group,the protein expression levels of Nrf2 downstream target genes,including NQO1 and HO-1,in the renal tissues of mice in the Zn group increased,and the levels of NQO1 and HO-1 in the renal tissues of mice in the DM/Zn group showed a further increase,with the differences being statistically significant(P<0.05).Compared to those of the mice in the Ctrl group,the protein expressions of Nrf2 downstream target genes,including SOD-1,SOD-2,and CAT of in the renal tissues of the mice in the Zn group increased,while the expression levels of SOD-1,SOD-2,and CAT in the renal tissues of the mice in the DM group decreased,with the differences being statistically significant(P<0.05).Zn supplementation could completely inhibit these changes(P<0.05).Conclusions Zn supplementation has therapeutic effects on DKD and mitigates T1DM-induced renal dysfunction and oxidative injury in mice,which may be associated with the activation of the Nrf2 antioxidant signaling pathway.

BACKGROUND: Electroacupuncture (EA) treatment is efficacious in patients with respiratory disorders, although the mechanisms of its action in lung-function protection are poorly understood. This study aimed to explore the neuroanatomical mechanisms of EA stimulation at the BL13 acupoint (Feishu, EA-BL13) improvement in asthma. METHODS: Allergic asthma was induced by intranasal 2.0% ovalbumin (OVA) instillation combined with intraperitoneal injection of the 10.0% OVA. The levels of interleukin (IL)-4 and IL-5 were detected by enzyme-linked immunosorbent assay. Hematoxylin and eosin and periodic acid-schiff stain were used to evaluate inflammatory cell infiltration and mucus secretion. Cellular oncogene fos induction in neurons after EA stimulation was detected by immunofluorescent staining. The messenger RNA expression levels of adrenergic receptors were quantified with real-time polymerase chain reaction. RESULTS: EA improved airway inflammation and mucus secretion mainly by activating somatosensory-sympathetic pathways ( P <0.001). Briefly, the intermediolateral (IML) nuclei of the spinal cord received signals from somatic EA stimulation and then delivered the information via the sympathetic trunk to the lung. Excited sympathetic nerve endings in lung tissue released large amounts of catecholamines that specifically activated the β2 adrenergic receptor (β2AR) on T cells ( P <0.01) and further decreased the levels of IL-4 and IL-5 ( P <0.001) through the cyclic adenosine monophosphate/protein kinase A signaling pathway. CONCLUSION This study provided a new explanation and clinical basis for the use of EA-BL13 as a treatment for allergic asthma in both the attack and remission stages and other respiratory disorders related to airway inflammation.
Electroacupuncture/methods* ; Animals ; Asthma/immunology* ; Rats ; Rats, Sprague-Dawley ; Male ; Inflammation/therapy* ; Interleukin-4/metabolism* ; Interleukin-5/metabolism*

Objective To study the regulatory effect of Qufeng Xiaodian Formula on serum differential metabolites of allergic purpura,and provide scientific basis for the diagnosis and treatment of allergic purpura in traditional Chinese medicine.Methods Sixty rats were randomly divided into a blank control group(referred to as the blank group),a model group,a compound glycyrrhizin group,a low-dose Qu Feng Xiao Dian Fang group,a medium dose Qu Feng Xiao Dian Fang group,and a high-dose Qu Feng Xiao Dian Fang group according to a random number table method,with 10 rats in each group.The model group was constructed by combining dried ginger,pepper,and long pepper with ovalbumin to create an allergic purpura rat model.After successful modeling,each treatment group was intervened with corresponding drugs for 4 weeks.After 4 weeks,serum was collected and non targeted metabolomics screening of serum differential metabolites was performed using ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometer(UPLC-QTOF/MS).Subsequently,data extraction and multivariate statistical analysis will be conducted to identify potential metabolic pathways.Results Compared with the control group,there were 91 possible differential metabolites in the serum of the model group rats,corresponding to 20 metabolic pathways;Compared with the model group,there were a total of 43 possible differential metabolites in the serum of rats in the wind dispelling and disease eliminating group,corresponding to 15 metabolic pathways.Among them,there are a total of 12 metabolic pathways.Inflammatory metabolites such as arachidonic acid and ceramide can damage vascular endothelium.Ten biomarkers,including arachidonic acid and ceramide,were significantly abnormal in the serum of the model group rats compared to the normal group.The Qufeng Xiaodian formula can significantly reverse these metabolites and significantly enrich them in arachidonic acid metabolism and sphingolipid metabolism pathways.Conclusion Qufeng Xiaodian Formula has a certain regulatory effect on metabolites such as arachidonic acid and ceramide that affect vascular endothelial injury.

Objective:To investigate the effect of rat platelet-rich plasma (PRP) gel on autologous adipose-derived mesenchymal stem cells (ADSCs) overexpressing glial-derived neurotrophic factor (GDNF).Methods:This study was an experimental study. Five adult male Sprague-Dawley rats were used, and the primary ADSCs were obtained by collagenase digestion, and then the cells were identified successfully. The 3 rd passage of ADSCs were obtained and divided into negative control group infected with unloaded adenovirus and overexpressing GDNF group infected with overexpressing GDNF adenovirus, according to random number table method (the grouping method was the same below). After 48 hours of culture, the infection of cells was observed. Five adult male Sprague-Dawley rats were used, and the PRP was obtained after collecting blood by differential centrifugation. PRP was prepared into a gel and its microstructure was observed by scanning electron microscope. The ADSCs of 3 rd passage were added into the PRP solution mixture and cultured for 48 hours after gelation. The cell growth was observed by hematoxylin-eosin staining and calcein/propyl iodide staining. ADSCs infected with unloaded adenovirus and ADSCs infected with overexpressing GDNF adenovirus were routinely cultured in PRP gel. After 48 hours of culture, the cell growth was detected by calcein/propyl iodide staining. After culture for 24, 48, 72 hours and 1, 2, 3, 4 weeks, the supernatant of cell culture medium was collected, the absorbance value was determined by microplate analyzer, and the GDNF content was calculated, with the sample number of 3. After 48 hours of culture, the expression of S100 protein (a specific marker of Schwann cells) was detected by immunofluorescence assay. Results:After 48 hours of culture, the proportions of cells infected with adenovirus in negative control group and overexpressing GDNF group were close to 90%, and the cell growth was good. The cells in negative control group grew normally. The morphology of the cells in overexpressing GDNF group changed significantly with 80%-90% of the cells having two or more protrusions, and the protrusions were interwoven to form a network wherever the cells gathered. PRP gel formed a three-dimensional network structure with different pore sizes. After 48 hours of culture, ADSCs could be well attached to PRP gel, and 98% of the cells were alive. After 48 hours of culture, ADSCs infected with unloaded adenovirus grew well and showed typical ADSC-like spindle-shaped growth. ADSCs infected with overexpressing GDNF adenovirus grew well, and most of the cells had two or more protrusions, and the protrusions were interwoven into a network. After culture for 24, 48, 72 hours and 1, 2, 3, 4 weeks, the content of GDNF in the supernatant of ADSCs infected with overexpressing GDNF adenovirus was (90±10), (133±15), (150±10), (137±15), (132±18), (120±10), and (127±16) pg/mL, which was significantly higher than (42±7), (44±7), (43±6), (47±6), (49±5), (49±6), and (51±4) pg/mL of ADSCs infected with unloaded adenovirus (with t values of 6.20, 8.08, 15.18, 9.12, 7.99, 9.61, and 7.86, respectively, P<0.05). After 48 hours of culture, the fluorescence intensity of S100 protein expression of ADSCs infected with overexpressing GDNF adenovirus was significantly stronger than that of ADSCs infected with unloaded adenovirus. Conclusions:The prepared autologous three-dimensional PRP gel has good biocompatibility and can carry rat GDNF-overexpressing ADSCs and release GDNF slowly, inducing ADSCs to differentiate into Schwann cells that express high level of S100 protein.

Objective To investigate the effects of DNA demethylation drugs combined with histone deacetylase in-hibitors on fragile X mental retardation 1 neighbor protein (FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression.Methods Human oral cancer cell lines Cal27 and SCC-9 were treated with decitabine (DAC) , an inhibitor of DNA meth-yltransferase, combined with trichostatin A (TSA) and valproic acid (VPA), inhibitors of histone deacetylase.Then reverse transcription-polymerase chain reaction (RT-PCR) , quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression of FMR1 NB and pyrosequencing was used to detect the methylation of FMR1NB promoter.Results Compared with the blank control group, DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in Cal27 and SCC-9 cells.Compared with DAC alone group, FMR1NB mRNA expression of each DAC-combined drug groups significantly increased, but FMR1NB protein did not significantly change in Cal27 cells; for SCC-9 cells, except for DAC+TSA group, the mRNA and protein levels of FMR1NB significantly increased in all other groups.In addition, there was no signifi-cant difference in the expression of FMR1 NB mRNA and protein between the three-combined drugs group and two-combined drugs groups.Further methylation assay showed that the methylation level of the overall FMR1NB promot-er and its each CpG site measured were reduced to varying degrees in all treatment groups except for three-combina-tion drug group of SCC-9.Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter, wherein the enhanced expression effect of the com-bination of the two drugs is stronger, suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.

Urine is produced from the urinary system, and urinary cell-free DNA (cfDNA) carries genomic DNA directly secreted from urinary system.Urine samples are non-invasive, unlimited in quantity and easy to obtain, making urinary cfDNA a promising biomarker for urologic diseases.This article reviews the progress of clinical application of urinary cfDNA in urologic diseases.