[Mechanism of Huangqin Qingre Chubi Capsules regulating p53/p21 signaling pathway to delay chondrocyte senescence in osteoarthritic rats].

This study aims to investigate the mechanism of Huangqin Qingre Chubi Capsules(HQC) in delaying chondrocyte senescence of osteoarthritic(OA) rats by regulating the p53/p21 signaling pathway. Rheumatic fever paralysis models of OA rats were induced based on monosodiun iodoacetate(MIA) combined with external rheumatic fever environmental stimuli and divided into normal(Con) group, OA model(MIA) group, OA model+rheumatic fever stimulation model(MIA-M) group, MIA-M+HQC low-dose(MIA-M+HQC-L) group, medium-dose(MIA-M+HQC-M) group, and high-dose(MIA-M+HQC-H) group, and MIA-M+glucosamine(MIA-M+GS) group. The models were successfully prepared and administered by gavage for 30 d. The pathological changes of cartilage were observed by hematoxylin-eosin(HE) and Senna O solid green(SO) staining. The expression of interleukin(IL)-1ß and IL-6 was detected by enzyme-linked immunosorbent assay(ELISA). Flow cytometry(FCM) was used to detect apoptosis and cell cycle. The mRNA expression of MMP13, ADAMTS-5, COLⅡ, and TGF-ß was detected by RT-qPCR. The protein expression of p53/p21, p16, Bax, and Bcl-2 was detected by Western blot. The articular cartilage surface of rats in the Con group was smooth, and the tide line was smooth. The cartilage layer of MIA and MIA-M groups was obviously damaged, and the cartilage matrix was reduced. The above conditions were more severe in the MIA-M group. The cartilage surface of the HQC high-dose group and MIA-M+GS group was basically intact with clear delamination. Compared with the MIA-M+HQC-H group, Mankin's score was higher in the HQC low-dose and medium-dose groups, and the change was not obvious in the MIA-M+GS group. Compared with the Con group, the proportion of chondrocytes G_1 was elevated in the MIA and MIA-M groups, and the proportion of the S phase and G_2 phase was significantly decreased. In addition, the apoptosis rate was increased. Compared with MIA-M, HQC groups inhibited apoptosis and promoted cell proliferation in a concentration-dependent manner. Compared with the MIA-M+HQC-H group, the effect was more significant in the HQC high-dose group than in the HQC medium-low dose, while it was not significant in the MIA-M+GS group. Compared with the Con group, IL-1ß and IL-6 were elevated in the MIA and MIA-M groups, and mRNA levels of MMP13 and ADAMTS-5 were elevated. p53, p21, p16, and Bax protein were elevated, and mRNA levels of COLⅡ and TGF-ß were decreased. Compared with the MIA-M group, IL-1ß and IL-6 decreased after drug interventions of HQC and GS, and mRNA levels of MMP13 and ADAMTS-5, as well as protein levels of p53, p21, Bax, and p16 decreased. In addition, Bcl-2 increased. The improvement of these indexes was significantly better in the MIA-M+HQC-H group than in the HQC low-dose and medium-dose groups, and the difference with the MIA-M+GS group was not significant. HQC delayed MIA-induced chondrocyte senescence in OA rats, inhibited inflammatory response and extracellular matrix(ECM) degradation, and its mechanism may be related to the inhibition of the p53/p21 pathway.

[Prescription and medication rules of traditional Chinese medicine for prevention and treatment of diabetic microangiopathy based on literature mining].

This study explored the prescription and medication rules of traditional Chinese medicine(TCM) in the prevention and treatment of diabetic microangiopathy based on literature mining. Relevant literature on TCM against diabetic microangiopathy was searched and prescriptions were collected. Microsoft Excel 2021 software was used to establish a prescription database, and an analysis was conducted on the frequency, properties, flavors, meridian tropism, and efficacy classifications of drugs. Association rule analysis, cluster analysis, and factor analysis were performed using SPSS Modeler 18.0 and SPSS Statistics 26.0 software. The characteristic active components and mechanisms of action of medium-high frequency drugs in the analysis of medication rules were explored through li-terature mining. A total of 1 327 prescriptions were included in this study, involving 411 drugs, with a total frequency reaching 19 154 times. The top five high-frequency drugs were Astragali Radix, Angelicae Sinensis Radix, Poria, Salviae Miltiorrhizae Radix et Rhizoma, and Rehmanniae Radix. The cold and warm drugs were used in combination. Drugs were mainly sweet, followed by bitter and pungent, and acted on the liver meridian. The majority of drugs were effective in tonifying deficiency, clearing heat, activating blood, and resolving stasis. Association rule analysis identified the highly supported drug pair of Astragali Radix-Angelicae Sinensis Radix and the highly confident drug combination of Poria-Alismatis Rhizoma-Corni Fructus. The strongest correlation was found among Astragali Radix, Angelicae Sinensis Radix, Poria, and Salviae Miltiorrhizae Radix et Rhizoma through the complex network analysis. Cluster analysis identified nine categories of drug combinations, while factor analysis identified 16 common factors. The analysis of active components in high-frequency drugs for the treatment of diabetic microangiopathy revealed that these effective components mainly exerted their effects by inhibiting oxidative stress and suppressing inflammatory reactions. The study found that the pathogenesis of diabetic microangiopathy was primarily characterized by deficiency in origin, with a combination of deficiency and excess. Deficiency was manifested as Qi deficiency and blood deficiency, while excess as phlegm-heat and blood stasis. The key organ involved in the pathological changes was the liver. The treatment mainly focused on supplementing Qi and nourishing blood, supplemented by clearing heat, coo-ling blood, activating blood, and dredging collaterals. Commonly used formulas included Danggui Buxue Decoction, Liuwei Dihuang Pills, Erzhi Pills, and Buyang Huanwu Decoction. The mechanisms of action of high-frequency drugs in the treatment of diabetic microangiopathy were often related to the inhibition of oxidative stress and suppression of inflammatory reactions. These findings can provide references for the clinical treatment of diabetic microangiopathy and the development of targeted drugs.

[Effect of Huangqin Qingre Chubi Capsules containing serum on oxidative stress and protein expression of AMPK and FoxO3a in rheumatoid arthritis patients].

To study the effect of Huangqin Qingre Chubi Capsules containing serum on the protein expressions of AMPK and FoxO3 a in peripheral blood mononuclear cells of patients with rheumatoid arthritis(RA), in order to explore the mechanism of anti-oxidation. Peripheral anticoagulant was collected from patients and normal people. Monocytes(PBMC) were isolated through density gradient centrifugation, and the logarithmic phase cells were cultured. Drug containing serum was prepared through intragastric admini-stration to SD rats. The rats were divided into five groups, namely normal group, model group, AMPK blocker group(compound C 10 µmol·L~(-1)), medium-dose HQC+AMPK blocker group, and middle-dose HQC group. The cell inhibition rate was calculated by MTT method. The levels of IL-1ß, IL-4, LPO, MDA, SOD and TAOC were detected by ELISA. The expressions of AMPK, p-AMPK, p-FoxO3 a and FoxO3 a were detected by Western blot. The HQC containing serum had an inhibitory effect on human monocytes in peripheral blood. The best concentration was observed in middle-dose HQC, and the best time was 24 hours. Middle-dose HQC group was better than model group, AMPK blocker group and middle-dose HQC + AMPK blocker group in terms of increase of SOD, p-AMPK, p-FoxO3 a and decrease of LPO. It was better than model group and AMPK blocker group in terms of increase of IL-4, TAOC, AMPK, FoxO3 a and decrease of IL-1ß, MDA. The differences were statistically significant(P<0.05 or P<0.01). The HQC containing serum may increase the levels of TAOC and SOD, decrease the level of MDA and LPO, activate AMPK, directly phosphorylate FOXO3 a, enhance its transcriptional activity, and improve the state of oxidative stress in RA patients.

[Regularity of prescription medication of Xin'an Wang's internal medicine in treating stomach cramps based on data mining].

The study analyzes the medication rules of Xin'an Wang's internal medicine for treating stomach cramps by data mining technology,in order to provide reference for clinical medication. Through the summarization of the medical cases of stomach cramps treated by Xin'an Wang's doctors( Wang Ren-zhi,Wang Zhong-qi,Wang Le-tao),statistics was made for the frequency of symptoms,signs,syndromes and drugs in Office 2010. Apriori algorithm in IBM SPSS Modeler 14. 1,and SPSS Statistics 22. 0 were used for association rule analysis and cluster analysis. The results showed that the 310 prescriptions collected involved totally 322 syndromes( including symptoms and signs) and 336 drugs,with the cumulative dose of 4 072 times; the symptoms were correlated to the spleen and stomach,liver and gallbladder,and the heart system; syndrome differentiation was mainly based on liver-Qi invasion of the stomach,diet impairment to the stomach,deficiency of spleen and stomach and cold syndrome; commonly used drugs were Qi regulating drugs,phlegm eliminating drugs and blood circulation promoting and stasis removing drugs; high-frequency drug complex network diagram showed that Pinelliae Rhizoma,Aurantii Fructus,Trichosanthis Fructus,Allii Macrostemonis Bulbus were closely related; the analysis showed 12,20,and 17 two,three,and four association rules; cluster analysis showed 10 pairs of Trichosanthis Fructus-Allii Macrostemonis Bulbus,Pinelliae Rhizoma-Aurantii Fructus,and Aspongopus-Toosendan Fructus drug combinations. According to Xin' an Wang's doctors,stomach cramps are closely related to liver and spleen,Qi stagnation,phlegm and blood stasis are the standard.Xin'an Wang's doctors give the first priority on " deoppilation",focus on soothing the liver and spleen,activating Qi and eliminating phlegm,and promoting blood circulation,and refer to use modified Xiaoxianxiong Decoction and modified Gualou Xiebai Banxia Decoction based on symptoms.

[Study on characteristics and data mining of 464 cases of lung diseases in Xin'an Wang's medicine].

The purpose of this paper is to study the use rules of drugs for lung diseases in internal medicine department of Xin'an Wang's family, discuss the compatibility of common drugs for lung diseases, guide clinical application, and inherit Xin'an medicine. By retrospective study on lung diseases cases in Wang's internal medicine works, the lung diseases and use frequency of common drugs treated by Wang's medicine were counted, and the systematic clustering and association rule analysis of common drugs were conducted by using SPSS Statistic 20 and SPSS Modeler 18.0, respectively. The results showed that asthma, cold and cough were the main lung diseases treated by Wang's medicine, and the commonly used medicines included antitussive and antiasthmatic drugs, spleen-invigorating and dampness-removing drugs, and expectorants. The medicine taste was mainly bitter, pungent and sweet, with cold and warm properties in a balanced way, without severely cold or hot herbs, mainly attributing to the lung and stomach meridians. In clustering analysis, 10 drug combinations were obtained; association analysis showed that two, three, four association rules respectively had 11, 21, and 10 groups, and each drug group had 11, 16, and 5 items. Core combinations: Poria, Armeniacae Semen Amarum, Asteris Radix et Rhizome, Coicis Semen, Farfarae Flos, Dendrobii Caulis, Perilla Frutescens, Stemonae Radix, Citri Reticulatae Pericarpium, Cynanchi Stauntonii Rhizome et Radix, Meretricis Concha Cyclinae Concha, Belamcandae Rhizoma, and Pinelliae Rhizome. Xin'an Wang's medicine paid attention to the lung nature when treating lung diseases. Lung is a delicate organ, not resistant to coldness or heat, so severely cold or hot herbs shall not be used, and the clear and light drugs with functions of dispersing lung Qi, clearing phlegm evil, strengthening spleen, eliminating phlegm, and relieving cough and asthma are often used. Lung deficiency is a kind of deficiency of Qi and Yin, so both Qi and Yin shall be regulated. Deficiency of Yin would burn the lung and make the lung collaterals blocked. In this case, the lung collaterals shall be dredged for hemostasis. Long time of lung deficiency would hinder the distribution of body fluid, and lung shall be regulated to dissipate phlegm.

Triptolide inhibits cell growth and inflammatory response of fibroblast-like synoviocytes by modulating hsa-circ-0003353/microRNA-31-5p/CDK1 axis in rheumatoid arthritis.

Triptolide (TPL) is an active component derived from Tripterygium wilfordii Hook F (TwHF) with therapeutic potential for rheumatoid arthritis (RA). However, the underlying mechanism of TPL is remains under-studied. Competing endogenous RNA (ceRNA) networks may participate in the response to TPL in RA. Herein, we sought to identify a TPL response-related ceRNA axis. A circular RNA (circRNA)-microRNA (miRNA)-mRNA ceRNA axis associated with the TPL response was constructed according to our previous study. Modulatory mechanisms of the ceRNA axis were ascertained through a series of experimentations. The clinical relevance of the ceRNA axis was also determined using computational models. Here, we found that TPL had excellent clinical effect on RA and promising therapeutic efficacy in experimental animals. The ceRNA axis of hsa-circ-0003353 (circ0003353), miR-31-5p, and CDK1 was identified as a candidate biomarker for the response of RA patients to TPL. TPL inhibited the viability, proliferation, and cell cycle entry of RA-fibroblast-like synoviocytes (FLSs), as well as the production of cytokines. Overexpression of circ0003353 abolished the inhibitory effects of TPL on RA-FLSs. Mechanistically, circ0003353 sponged miR-31-5p that inversely targeted CDK1 and manipulated the p21/Cyclin B axis. Additionally, consecutive rescue experiments indicated that the inhibitory impacts of TPL on RA-FLSs were dependent on the circ0003353/miR-31-5p/CDK1 axis. Molecular docking was also applied to predict the specific binding sites and binding capacity of TPL to related targets. In conclusion, the present study demonstrated that TPL repressed the cell growth and inflammatory response of RA-FLSs by mediating the expression of the circ0003353/miR-31-5p/CDK1 axis. This novel ceRNA axis may serve as a biomarker for screening RA patients who respond to TPL treatment, which holds potential applications in the diagnosis and therapy of RA.

Two protein-coding genes act as a novel clinical signature to predict prognosis in patients with ovarian serous cystadenocarcinoma.

Ovarian cancer is the seventh most common type of cancer and the eighth most common cause of cancer-associated mortality among women. A number of studies have hypothesized that the expression status of certain genes may be used to predict prognosis in ovarian cancer. In the present study, the RNA expression data from next-generation sequencing and the clinical information of 413 patients from The Cancer Genome Atlas dataset was downloaded to identify the association between gene-expression level and the survival time of the patients with ovarian serous cystadenocarcinoma. A five-gene model was predicted to be significantly associated with patient survival in ovarian serous cystadenocarcinoma by using random survival forests variable hunting algorithm and Cox analysis. A total of two genes, mesencephalic astrocyte-derived neurotrophic factor and dedicator of cytokinesis 11, of the predicted five genes demonstrated positive expression in the ovarian serous cystadenocarcinoma cancer tissues by polymerase chain reaction analysis. Kaplan-Meier and Receiver Operating Characteristic analysis confirmed that the model of the two genes exhibited high sensitivity and specificity to predict the prognostic survival of patients. In conclusion, the expression of the two genes in the two-gene model was associated with the prognostic outcomes of patients with ovarian serous cystadenocarcinoma; the model demonstrated potential as a novel prognostic indicator, which may have important clinical significance.