ObjectiveTo explore the potential mechanisms of action of the modified Duhuo Jisheng Mixture （JDJM） in treating synovial lesions in knee osteoarthritis （KOA）. MethodsA total of 43 male New Zealand white rabbits were randomly allocated into a blank group （n=8） and a model group （n=35）. The KOA model was induced by immobilizing the right hind limb with a high-molecular resin plaster bandage， with a modeling period of 6 weeks， resulting in successful modeling in 32 rabbits. These rabbits were then randomly allocated to the model group， celecoxib group， JDJM group and JDJM+740Y-P group， each consisting of 8 rabbits. The celecoxib group received celecoxib via gavage at a single dose of 0.009 3 g·kg^-1， while the JDJM was administered a single dose of 6.8 mL·kg^-1 （4.515 2 g·kg^-1） of the herbal preparation via gavage. The phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） pathway activator + JDJM group received 4.515 2 g·kg^-1 of the herbal preparation via gavage along with an auricular vein injection of 0.15 μmol·kg^-1 740Y-P. For a period of 6 weeks， the remaining groups received an equal volume of physiological saline via gavage daily. After the medication period， the knee joint pain threshold and circumference were measured， and hematoxylin-eosin （HE） staining was performed to assess the pathological changes in the synovial tissues. Enzyme-linked immunosorbent assay （ELISA） measured the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-18） and tumor necrosis factor-α （TNF-α） in the joint fluid. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to assess the mRNA expression of PI3K， Akt， mTOR， NOD-like receptor protein 3 （NLRP3）， cysteine-requiring aspartate protease-1 （Caspase-1） and gasdermin D （GSDMD） in the synovial tissues. Immunohistochemical （IHC） assay was performed to assess the protein expression of NLRP3， Caspase-1 and GSDMD. Western blot was carried out to analyze the protein expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， NLRP3， Caspase-1 and GSDMD. ResultsCompared to the blank group， the model group showed a significant increase in knee joint circumference and decrease in pain threshold， the synovial tissue pathology score was higher （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid significantly increased （P<0.01）. PI3K， Akt， mTOR phosphorylation as well as mRNA and protein expression increased （P<0.01）， while the mRNA and protein expression levels of NLRP3， Caspase-1 and GSDMD also significantly increased （P<0.01）. Compared to the model group， the celecoxib and JDJM groups exhibited a significant reduction in knee joint circumference and increase in pain threshold， the synovial tissue pathology score was lower （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid decreased （P<0.01）. The mRNA and protein expression of p-PI3K， p-Akt， p-mTOR， NLRP3， Caspase-1 and GSDMD were reduced （P<0.01）. Compared to the JDJM group， the JDJM+740Y-P group showed a decrease in the improvement of synovial lesions， an increase in knee joint circumference， and a decrease in pain threshold. The synovial tissue pathology score was lower （P<0.05）， and the levels of IL-1β， IL-18， and TNF-α in the joint fluid were higher （P<0.01）. The mRNA and protein expression of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR， NLRP3， Caspase-1 and GSDMD increased （P<0.01）. ConclusionJDJM is effective in treating KOA. Its mechanism may involve modulating the PI3K/Akt/mTOR pathway in synovial tissues， inhibiting pyroptosis， reducing inflammatory factor release， and protecting bony structures.

A method for determination and targeted screening of diflorasone components in health products using ultra performance liquid chromatography-quadrupole time of flight mass spectrometry(UPLC-Q-TOF/MS)was established.Four representative diflorasone and esters(diflorasone,diflorasone diacetate,diflorasone-17-propionate,and diflorasone-21-propionate)were selected to optimize the pretreatment conditions,and 10 mL of extraction solvent dosage,15 min of extraction time and 5 g of salting-out agent as the optimal conditions were selected by response surface methodology.The results showed that the four analytes exhibited good linearity within the concentration range of 2.0?100 μg/L with the chromatographic peak area,and the correlation coefficients(R2)were all greater than 0.9990,while the results of recovery and relative standard deviation could satisfy the requirements of determination.The common characteristic ions of diflorasone and esters werem/z121 andm/z335,and their specific structures were obtained by analyzing the cleavage pathway based on the optimized determination conditions.A targeted screening method for other esters of diflorasone based on characteristic ions guidance strategy was established.This method had many advantages such as high efficiency,high sensitivity and good reproducibility,and could be used for targeted screening of diflorasone and esters in health products.The developed characteristic ion guided strategy could be employed to construct mass spectral databases for various glucocorticoids,enabling comprehensive targeted screening across a broad range of compounds.

Objective To investigate the clinical characteristics,distribution of traditional Chinese medicine(TCM)syndrome types,and the anxiety and depression status in patients with colorectal polyps.Methods From January 2022 to December 2023,a total of 200 cases of patients with colorectal polyps(polyp group)and 100 patients without intestinal abnormalities(non-polyp group)under colonoscope who received colonoscopy or treatment at the Endoscopy Center of Dongguan Hospital,Guangzhou University of Chinese Medicine were enrolled.The relevant medical information of the patients in the two groups were collected,and then TCM syndrome differentiation of patients with colorectal polyps were carried out,the general infromation including gender,age,body mass index(BMI),smoking history and alcohol-intake history was recorded,and Hospital Anxiety and Depression Scale(HAD)score was calculated for assessing their anxiety and depression status.Results(1)The investigation of clinical characteristics showed that between the polyp group and the non-polyp group,the differences in the general information of gender,age,BMI,smoking history and alcohol-intake history were statistically significant(P<0.05 or P<0.01).As for the gender,the polyp group was predominated by the male(63.5%),while the non-polyp group was predominated by the female(58.0%),indicating that male patients were more likely to suffer from colorectal polyps.In terms of the age,the age group of 51-60 years old accounted for a higher percentage(36.0%)in the polyp group,while the age group of 41-50 years old accounted for a higher percentage(38.0%)in the non-polyp group,indicating that the polyp group had an elder average age than the non-polyp group.In addition,the percentages of patients with obesity(BMI≥28.0 kg/m2),smoking history,and alcohol-intake history in the polyp group were higher than those in the non-polyp group.(2)The analysis of features of colorectal polyps showed that the polyps were mostly seen in the left colon,accounting for 53.5%in the polyp group;adenomatous polyps accounted for 57.0%of all polyps;the size of colorectal polyps was usually less than one centimeter,accounting for 81.5%of all polyps;single colorectal polyps accounted for 45.5%,and multiple colorectal polyps accounted for 54.5%,indicating that colorectal polyps were usually multiple.(3)Among the 200 patients with colorectal polyps,there were 93 cases of syndrome of stagnation dampness due to spleen deficiency(46.5%),59 cases of dampness-heat in large intestine syndrome(29.5%),27 cases of liver depression and qi stagnation syndrome(13.5%),and 21 cases of dampness and blood stasis obstruction syndrome(10.5%).There were no statistically significant differences in the distribution of TCM syndrome types among colorectal polyp patients with different genders,ages,BMIs and alcohol consumption(P>0.05).The difference in the distribution of TCM syndrome types among colorectal polyp patients with or without smoking history was statistically significant(P<0.05).No statistically significant differences were shown in the distribution of TCM syndrome types among the colorectal polyp patients with various foci,number of polyps,and pathological types(P>0.05).The difference in the distribution of TCM syndrome types in colorectal polyp patients with different polyp sizes was statistically significant(P<0.05).In the patients with the polyp size0.05).In addition,the differences of anxiety scores and depression scores in the patients with various TCM syndrome types were not statistically significant(P>0.05).Conclusion(1)The distribution of TCM syndrome types of colorectal polyps is predominated by the syndrome of stagnation dampness due to spleen deficiency and dampness-heat in large intestine syndrome.The distribution of TCM syndrome types has no correlation with anxiety and depression,and the polyp foci,number of polyps,and pathological types of polyps,but has certain correlation with the size of the polyps.(2)Anxiety and depression may be the risk factors for colorectal polyps,but the polyp foci,number of polyps,and pathological types of polyps have no correlation with anxiety and depression,and the size of polyps may have certain correlation with anxiety.(3)Gender,age,smoking history and alcohol-intake history may be the risk factors for colorectal polyps.

Objective To observe the clinical efficacy of oral use combined with targeted transdermal delivery of Osteoking in treating knee osteoarthritis(KOA)of cold-damp blockage type.Methods A total of 120 patients with KOA of cold-damp blockage type who admitted to Hunan Provincial Hospital of Integrated Traditional Chinese And West Medicine from September 2022 to September 2023 were randomly divided into the control group,oral use group,transdermal delivery group and combination group according to the random number table method,with 30 cases in each group.The control group was treated with Celecoxib Capsule orally,the oral use group was treated with Osteoking orally,the transdermal delivery group was treated with Osteoking by targeted transdermal delivery,and the combination group was treated with oral use combined with targeted transdermal delivery of Osteoking.One course of treatment covered 12 days,and all of the four groups were treated for two continuous courses.Before and after treatment,the scores of traditional Chinese medicine(TCM)syndrome,Visual Analogue Scale(VAS)for pain,Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)for joint function,36-Item Short Form Health Survey(SF-36)for quality of life(QOL),as well as the levels of serum interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factor α(TNF-α)were observed in the patients of each group.After treatment,the clinical efficacy and safety of patients in each group were evaluated.Results(1)After two courses of treatment,the total effective rate of the combination group was 96.67%(29/30),which was higher than those of the control group[73.33%(22/30)],oral use group and transdermal delivery group[both being 70.00%(21/30)],and the difference was statistically significant(P<0.05).However,no statistically significant difference of the total effective rate was presented among the control group,oral use group,and transdermal delivery group(P>0.05).(2)After treatment,the scores of TCM syndrome,VAS for pain and WOMAC for joint function in the four groups were decreased compared with those before treatment(P<0.01).The intergroup comparison showed that the scores of TCM syndrome,VAS for pain and WOMAC in the combination group were lower than those in the other three groups(P<0.05),while no statistically significant difference of the scores of TCM syndrome,VAS for pain and WOMAC were presented among the control group,oral use group,and transdermal delivery group(P>0.05).(3)After treatment,the QOL scores of the eight dimensions of SF-36 such as physical functioning(PF),bodily pain(BP),general health(GH),vitality(VT),role-physical(RP),social functioning(SF),role-emotional(RE),and mental health(MH)in the four groups were significantly increased compared with those before treatment(P<0.01).The intergroup comparison showed that the scores of each dimension of SF-36 in the combination group were significantly higher than those in the other three groups(P<0.05),while no statistically significant difference of the dimension score of SF-36 was presented among the control group,oral use group,and transdermal delivery group(P>0.05).(4)After treatment,the levels of serum inflammatory factors of IL-1β,IL-6,and TNF-α in the four groups were decreased compared with those before treatment(P<0.01).The intergroup comparison showed that the decrease of serum levels of inflammatory factors in the combination group were significantly superior to those in the other three groups(P<0.05),and the decrease in the control group and oral use group were all superior to those in the transdermal delivery group(P<0.05),while the difference between the control group and oral use group was not statistically significant(P>0.05).(5)During the trial,no serious adverse reactions were found in the patients of four groups,which is of high safety.Conclusion Oral use combined with targeted transdermal delivery of Osteoking is effective on improving the joint pain,joint function and QOL of patients with KOA of cold-damp blockage type,and can speed up the rehabilitation of patients.Its therapeutic mechanism may be related to the decrease of the expression level of inflammatory factors.

Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there is still a lack of high-throughput methods for screening protein regulators of LLPS in target proteins. Here, we developed a CRISPR/Cas9-based screening method to identify protein phase separation regulators by integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly developed method, we screened the RNA-binding proteins that regulate PABPN1 phase separation and identified the tumor suppressor QKI as a promoter of PABPN1 phase separation. Furthermore, QKI exhibits decreased expression levels and diminished nuclear localization in colorectal cancer cells, resulting in reduced PABPN1 phase separation, which, in turn, promotes alternative polyadenylation (APA), cell proliferation, and migration in colorectal cancer.
Humans ; Colorectal Neoplasms/genetics* ; RNA-Binding Proteins/genetics* ; Poly(A)-Binding Protein I/genetics* ; CRISPR-Cas Systems ; Flow Cytometry ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

Objective: To study the impact of disaggregation on the quality of in vitro aggregated apheresis platelets. Methods: The apheresis platelets collected from Guangzhou Blood Center served as the study samples. The in vitro aggregated apheresis platelets after successful disaggregation were designated as the experimental group (referred to as the aggregation group), and apheresis platelets without in vitro aggregation during collection served as the control group. The product volume, platelet content, residual white blood cells, red blood cell contamination, pH value, CD62p expression rate, platelet morphology score and thromboelastography of both groups were respectively detected. Results: The routine quality control indicators such as product volume, platelet content, residual white blood cells, red blood cell contamination, and pH value of both groups all met the quality requirements. There were statistically significant differences in pH value (7.180 vs 7.071) between the two groups (P<0.05). There was no significant difference in product volume, platelet content, residual white blood cells, and red blood cell contamination between the two groups (P>0.05). The CD62p expression rate of the aggregation group was higher than that of the control group (42.386% vs 17.310%, P<0.05), while the cell morphology score of the aggregation group was lower than that of the control group (132.066 vs 141.166, P<0.05). No statistically significant differences were found in thromboelastography parameters (R-CK, K-CK, α angle, MA-CK, CI-CK) between the two groups (P>0.05). Conclusion: After the disaggregation of in vitro-aggregated apheresis platelets, the quality indicators met the national quality requirements. Although the expression rate of CD62p increased and the cell morphology score decreased, there were no statistically significant differences in the thromboelastography parameters between the two groups. This indicates that although some platelet activation occurred, it did not affect the hemostatic function of the platelets.

OBJECTIVES: To compare the anti-inflammatory, antibacterial and analgesic effects of Yinqiao Powder (YQS) formulated granules and decoction. METHODS: We first evaluated the anti-inflammatory effects of the two dosage forms of YQS in a LPS-induced RAW 264.7 cell model using RT-qPCR and Western blotting. We further constructed zebrafish models of inflammation by copper sulfate exposure, caudal fin transection, or LPS and Poly (I:C) microinjection, and evaluated anti-inflammatory effects of YQS granules and decoction by examining neutrophil aggregation and HE staining findings. In a mouse model of acute lung injury (ALI) induced by intratracheal LPS instillation, the effects of YQS gavage at 10, 15, and 20 g/kg on lung pathologies were evaluated by calculating lung wet-dry weight ratio and using HE staining, ELISA and Western blotting. The microbroth dilution method was used to evaluate the antibacterial effect of YQS. Mouse pain models established by hot plate and intraperitoneal injection of glacial acetic acid were used to evaluate the analgesic effects of YQS at 10, 15, and 20 g/kg. RESULTS: Both YQS granules and decoction significantly reduced TNF-α, IL-6, and IL-1β expressions and p-STAT3 (Tyr 705) phosphorylation level in LPS-induced RAW 264.7 cells, and obviously inhibited neutrophil aggregation in the zebrafish models. In ALI mice, YQS granules and decoction effectively ameliorated lung injury, lowered lung wet-dry weight ratio, and reduced p-STAT3 (Tyr 705) expression and TNF-α and IL-6 levels. YQS produced obvious antibacterial effect at the doses of 15.63 and 31.25 mg/mL, and significantly reduced body torsion and increased pain threshold in the mouse pain models. CONCLUSIONS The two dosage forms of TQS have similar anti-inflammatory, antibacterial and analgesic effects with only differences in their inhibitory effect on TNF-α, IL-6 and IL-1β mRNA expressions in LPS-induced RAW 264.7 cells.
Animals ; Mice ; Drugs, Chinese Herbal/pharmacology* ; Anti-Inflammatory Agents/pharmacology* ; Analgesics/pharmacology* ; RAW 264.7 Cells ; Zebrafish ; Anti-Bacterial Agents/pharmacology* ; Powders ; Tumor Necrosis Factor-alpha/metabolism* ; Acute Lung Injury/drug therapy* ; Interleukin-6/metabolism* ; Lipopolysaccharides

Objective To analyze the imaging features of cerebral small vessel disease in patients with type 2 diabetes mellitus by multimodal MRI.Methods The clinical data of 160 patients with cerebral small vessel disease admitted to our hospital from January to December 2020 were retrospectively analyzed.According to whether they were complicated with type 2 diabetes mellitus,they were divided into the diabetic group and the non-diabetic group,with 80 cases in each group.Both groups underwent multimodal MRI scans.And the severity of lacunar infarction,the severity of subcortical and periventricular white matter lesions,white matter integral and cerebral microbleeds of patients in the two groups were compared.Results The severity of lacunar infarction(χ2=34.076,P=0.001),subcortical white matter lesions(χ2=25.000,P=0.001),periventricular white matter lesions(χ2=22.895,P=0.001)and white matter integral(t=12.370,P=0.001)of patients in the diabetic group were significantly higher than those in the non-diabetic group.No cerebral microbleeds were detected in either group of patients.Conclusion Patients with cerebral small vessel disease and type 2 diabetes mellitus show characteristic multimodal MRI changes.The increase in the number of lacunar infarction lesions and the aggravation of white matter lesions can be used as the characteristic imaging basis for the diagnosis of type 2 diabetes mellitus related cerebral small vessel disease.

AIM To explore the clinical effects of Supplemented Buyang Huanwu Decoction on postoperative patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern.METHODS One hundred and twenty patients were randomly assigned into control group(60 cases)for 6-week intervention of conventional treatment,and observation group(60 cases)for 6-week intervention of both Supplemented Buyang Huanwu Decoction and conventional treatment.The changes in clinical effects,TCM syndrome scores,spinal cord conduction signals(SEP amplitude,MEP amplitude),serum neurotrophic factors(NGF,IGF-1,BDNF),coagulation and inflammatory indices(PT,APTT,TNF-α,IL-1 β)and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased TCM syndrome scores,TNF-α,IL-1β(P＜0.05),increased spinal cord conduction signals,coagulation and inflammatory indices(P＜0.05),and shortened PT,APTT(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern,Supplemented Buyang Huanwu Decoction can safely and effectively promote neurological function recovery.

Objective:To analyze serum inflammatory factors associated with antihistamine resistance in patients with chronic spontaneous urticaria (CSU) .Methods:A total of 88 CSU patients were enrolled from Guangzhou Dermatology Hospital from January 2022 to December 2024. All patients received antihistamine treatment according to the "Guideline for diagnosis and treatment of urticaria in China (2022) " . Based on the 7-day urticaria activity score (UAS7) after 4-week treatment, these patients were divided into an antihistamine-sensitive group and an antihistamine-resistant group. Serum levels of inflammatory factors at the initial visit were analyzed using the Olink-targeted proteomics technology. Specific biomarkers associated with antihistamine resistance were identified, and Spearman correlation analysis was carried out to analyze correlations among differentially expressed proteins. A logistic regression model was constructed based on the Olink proteomics data, and the predictive performance of the model was evaluated using receiver operating characteristic (ROC) curve analysis. Measurement data were expressed as mean ± standard deviation or median (lower quartile, upper quartile) .Results:The 88 CSU patients aged 12 to 81 (38.78 ± 13.89) years, with the disease duration being 18 (7.00, 60.00) months. There were 32 patients in the antihistamine-sensitive group and 56 in the antihistamine-resistant group. No significant differences were found between the two groups in terms of age, disease duration, gender, or history of allergic diseases (all P > 0.05) . After 4 weeks of antihistamine treatment, the UAS7 score was significantly higher in the antihistamine-resistant group (25.00 [15.25, 31.00] points) than in the antihistamine-sensitive group (0.50 [0.00, 4.00] points; Z = -7.08, P < 0.001) . The Olink-targeted proteomics identified 5 differentially expressed proteins between the two groups: compared with the antihistamine-sensitive group, the antihistamine-resistant group showed > 2-fold higher expression of fibroblast growth factor 19 (FGF19) , interleukin-15 receptor subunit alpha (IL-15RA) , eotaxin (CCL11) , and monocyte chemoattractant protein-1 (MCP-1) ; in contrast, the expression of sulfotransferase 1A1 (ST1A1) in the antihistamine-sensitive group was 2.54 times that in the antihistamine-resistant group. Among the differentially expressed proteins, MCP-1 showed the highest specificity (1.00) for predicting antihistamine resistance, followed by CCL11 (0.97) . Correlation analysis revealed a significant positive correlation between MCP-1 and CCL11, and a significant negative correlation between IL-15RA and ST1A1. ROC curve analysis showed that MCP-1 and CCL11 had area under the curve values of 0.603 and 0.630, respectively, in predicting antihistamine resistance. Conclusion:MCP-1 and CCL11 may be potential biomarkers for predicting antihistamine resistance in CSU patients.