OBJECTIVES: To assess the efficacy and safety of Tiaozhou Ziyin (TZZY) recipe for treatment of premature ovarian insufficiency (POI) and explore the possible mechanisms. METHODS: We used bioinformatics analyses and network pharmacology to identify the main active ingredients in TZZY recipe and their core targets, which were verified by Western blotting. We tested the efficacy and safety of the recipe in 60 POI patients, who were randomized into control group (n=30) with Femoston treatment and TZZY group (n=30) with additional TZZY recipe treatment for 3 menstrual cycles. RESULTS: The core active ingredients of TZZY recipe included kaempferol, β-sitosterol, luteolin, and quercetin. The core targets included SRC, TP53, STAT3, PIK3CA, and MAPK3, which were involved in positive regulation of cell movement and protein phosphorylation, the cancer pathways and the PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had good binding ability with the core targets. In female rat models of POI, TZZY recipe treatment significantly up-regulated ovarian expressions of p-PI3K and p-Akt proteins. In the clinical trial, treatment with Femoston and Femoston plus TZZY recipe both significantly increased E₂ levels and reduced FSH and LH levels and Kupperman scores of the patients, and the combined treatment produced significantly stronger effects. Both treatments increased the number of antral follicles of the patients, but the combined treatment also significantly increased the levels of AMH. CONCLUSIONS The therapeutic mechanism of TZZY recipe for POI involves multiple active ingredients, multiple therapeutic targets and multiple pathways, and activating the PI3K /Akt pathway is one of its main mechanisms of action, to improve ovarian reserve function, alleviate clinical symptoms, and enhance clinical efficacy in POI patients.
Female ; Primary Ovarian Insufficiency/drug therapy* ; Humans ; Drugs, Chinese Herbal/therapeutic use* ; Animals ; Rats ; Molecular Docking Simulation ; Signal Transduction ; Sitosterols/therapeutic use* ; Kaempferols/therapeutic use*

Objective To explore the establishment of a prognostic model based on machine learning algorithm to predict primary graft dysfunction(PGD)in patients with idiopathic pulmonary fibrosis(IPF)after lung transplantation.Methods Clinical data of 226 IPF patients who underwent lung transplantation were retrospectively analyzed.All patients were randomly divided into the training and test sets at a ratio of 7∶3.Using regularized logistic regression,random forest,support vector machine and artificial neural network,the prognostic model was established through variable screening,model establishment and model optimization.The performance of this prognostic model was assessed by the area under the receiver operating characteristic curve(AUC),positive predictive value,negative predictive value and accuracy.Results Sixteen key features were selected for model establishment.The AUC of the four prognostic models all exceeded 0.7.DeLong and McNemar tests found no significant difference in the performance among different models(both P＞0.05).Conclusions Based on four machine learning algorithms,the prognostic model for grade 3 PGD after lung transplantation is preliminarily established.The overall prediction performance of each model is similar,which may predict the risk of grade 3 PGD in IPF patients after lung transplantation.

Objective:To explore the differences of gene expression profiles of precursors of exhausted T cells (Tpex) and terminal exhausted T cells (Tex) in the peripheral blood of patients with active pulmonary tuberculosis (ATB).Methods:Twenty-five cases of ATB, 13 cases of latent tuberculosis infection (LTBI) and 10 health controls were enrolled from January 2021 to October 2022 in the Fifth People′s Hospital of Wuxi. The proportions of Tpex and Tex in the peripheral blood mononuclear cells (PBMCs) of the three groups were detected by flowcytometry. PBMCs of ATB were separated into Tpex and Tex by fluorescence-activated cell sorting. RNA-sequencing was performed and up-regulated and down-regulated genes were screended. Differently expressed genes were analyzed by gene set enrichment analysis of gene ontology (GO) to find regulatory pathways affecting cell metabolism and function. Wilcoxon matched-pairs signed rank test, Kruskal-Wallis test and Dunn multiple comparsion test were used for statistical analysis.Results:The proportion of Tpex in ATB group was 2.86%(1.74%), which was lower than 7.93%(6.16%) of Tex, and the difference was statistically significant ( Z=-3.91, P<0.001). The proportions of Tpex and Tex in LTBI group were 9.47%(6.26%) and 7.43%(5.48%), respectively, and the difference was not statistically significant ( Z=-0.93, P=0.345). The proportions of Tpex and Tex in healthy control group were 8.42%(2.69%) and 6.49%(5.14%), respectively, with no statistical significance ( Z=-1.36, P=0.170). There was statistical difference of the proportion of Tpex among the three groups ( H=21.93, P<0.001), and the proportion of Tpex in ATB group was lower than those in LTBI and heathy control groups, and the differences were both statistically significant ( Z=4.16, P<0.001 and Z=3.34, P=0.003, respectively), while the proportions of Tex in these three groups were not statistically different ( H=2.17, P=0.338). Compared with Tex, the gene expressions of memory markers, such as B-cell lymphoma 2 of Tpex were up-regulated, and the gene expressions of exhausted markers, such as lymphocyte activation gene 3 were down-regulated. In terms of cellular metabolism, the gene expressions of mitochondrial protein complex, mitochondrial matrix and oxidative phosphorylation of Tpex were up-regulated, and the gene expressions of glycolysis were down-regulated. The gene expressions of pyruvate metabolism in Tex were up-regulated, and the gene expressions of CD4 + T lymphocyte activation and differentiation and glycolytic process in Tpex were down-regulated. Conclusions:Tpex in ATB express more characteristics of memory cells and less features of exhausted markers compared with Tex, and the function of mitochondria of Tpex preserves well.

RNA ; CRISPR-Cas Systems

Objective : To study the effect of crisaborole on imiquimod (IMQ) Ⅳinduced psoriasis in mice. Methods: Forty eight Balb/c mice were randomly divided into crisaborole group (7. 5 , 15 , 30 mg/cm2），halometasone group ( 15 mg/cm2 ) , model group and normal group. IMQ was applied to the back of mice to establish the psoriasis model. Psoriasis area and severity index ( PASI) score was calculated , pathological changes , skin epidermal thickness and inflammatory cell infiltration in the dermis were observed by HE staining. The expressions of keratin (K) 1 , K10 , K6 , K16 and K17 in skin lesions were detected by Western blot and immunohistochemistry. The levels of cyclic adenosine monophosphate ( cAMP) , protein kinase A ( PKA) and phospho⁃cAMP response element binding protein (p⁃CREB) were detected. Results : Compared with the model group , the PASI score of the crisaborole group decreased , the expression levels of proliferative keratin ( K6 , K16 and K17 ) decreased( F = 12. 62、19. 41、28. 39 ,P < 0. 01) , and the expression levels of differentiation keratin (K1 and K10) increased(F = 27. 95、9. 64 , P < 0. 01) . Conclusion Crisaborole plays a therapeutic role in IMQ⁃induced psoriasis in mice by regulating the abnormal proliferation and differentiation of keratinocytes.

Purpose: To determine whether microRNA could be a therapy target of erectile dysfunction (ED) and the underlying mechanisms. Materials and Methods: Eight-week-old fasting male SD rats were intraperitoneally injected with streptozotocin to construct diabetic rat models. Diabetic ED rats were treated with miRNA-92a inhibitor. The cavernous nerves were electrically stimulated to measure the intracavernous pressure and mean arterial pressure of rats in each group. After the detection, the penile cavernous tissues are properly stored for subsequent experiments. Rat aortic endothelial cells were used in in vitro studies. Results: The expression of miR-92a was significantly increased in the corpus cavernosum of Streptozocin (STZ)-induced diabetic rats and injection of miR-92a antagomir into the corpus cavernosum of diabetic rats significantly increased eNOS/NO/ cGMP signaling pathway activities, cavernous endothelial cell proliferation, endothelial cell-cell junction protein expression and decreased the levels of oxidative stress. These changes restored erectile function in STZ-induced diabetic rats. Moreover, in vitro study demonstrated that the miR-92a expression increased significantly in endothelial cells treated with high glucose, inhibiting AMPK/eNOS and AMPK/Nrf2/HO-1 signaling pathways in rat aortic endothelial cells via targeting Prkaa2, causing endothelial dysfunction and overactive oxidative stress, miR-92a inhibitor can improve the above parameters. Conclusions miRNA-92a inhibitor could exert an inhibition role on oxidative stress and endothelial dysfunction to improve diabetic ED effectively.

Objective To explore the regulatory effect of miR-26b-5p on PI3K/PIP3 signaling path-way and its effect on L-type calcium channel of atrial cardiomyocytes in rats with ischemic ar-rhythmia.Methods A total of 72 SD rats were randomly divided into sham operation group,mod-el group,LY294002 group,negative control group,overexpression group and combination group,with 12 rats in each group.In 24 h after corresponding intervention,rat model of ischemic arrhyth-mia was established in the other groups except the sham operation group(only thoracotomy but without ligation).During the modeling process,arrhythmia indexes were detected in each group,and Curtis-Walker scoring was used for arrhythmia score.Fluorescence quantitative PCR was em-ployed to detect the expression of miR-26b-5p in atrial myocardiocytes,whole-cell patch-clamp technique was used to record the ICa-L of atrial myocardiocytes,and Western blotting was applied to measure the expression of CACNA1C,calmodulin and PI3K/PIP3 pathway related proteins.Re-sults Compared with the model group,the number of left ventricular preventricular contrac-tions,duration of ventricular tachycardia,duration of ventricular fibrillation,arrhythmia score,ab-solute value of ICa-L peak density of atrial myocardiocytes,and expression levels of CACNA1C and calmodulin were significantly increased,and the levels of miR-26b-5p,p-PI3K,PIP3 and p-Akt were obviously decreased in the LY294002 group(P＜0.05).While the overexpression group had decreased number of left ventricular preventricular contractions,shorter duration of ventricular tachycardia,shorter duration of ventricular fibrillation,lower arrhythmia score,reduced absolute value of ICa-L peak density,and lower expression levels of CACNA1C and calmodulin,and in-creased levels of miR-26b-5p,p-PI3K,PIP3 and p-Akt than the model group(P＜0.05).The number of left ventricular preventricular contractions,duration of ventricular tachycardia,dura-tion of ventricular fibrillation,arrhythmia score,absolute value of ICa-L peak density and expres-sion levels of CACNA1C and calmodulin were significantly increased,and the expression of p-PI3K,PIP3 and p-Akt were obviously decreased(0.82±0.08 vs 1.09±0.11,0.91±0.09 vs 1.17± 0.11,0.94±0.09 vs 1.20±0.12,P＜0.05)in the combination group than the overexpression group.Conclusion Overexpression of miR-26b-5p may block the L-type calcium channel in atrial myocardiocytes in ischemic arrhythmia rats by activating PI3K/PIP3 related signaling pathway,and thus improve the performance of arrhythmia in rats.

Humans ; Pilot Projects ; Prospective Studies ; Pulmonary Emphysema/surgery* ; Lung/surgery* ; Emphysema ; Perfusion ; Tomography, Emission-Computed ; Tomography ; Tomography, Emission-Computed, Single-Photon ; Pneumonectomy ; Bronchoscopy/methods*

Objective To investigate the potential risk factors for mild cognitive impairment (MCI) in the elderly population in the community, and to provide a basis for the primary prevention of MCI. Methods A cross-sectional study of elderly population in communities of Shanghai, China was conducted. A total of 368 subjects including both males and females, aged 65-80 years old, were selected to complete the mini-mental state examination (MMSE), basic information questionnaires, and physical examinations. Logistic regression analysis was used to analyze the potential risk factors of MCI. Results Of the 368 subjects participating in the study, 53 were found to have MCI and the prevalence rate was 14.4%. Univariate analysis found that older age, low education, no folic acid supplementation, stroke, osteoporosis and hyperlipidemia were risk factors of MCI. Multiple logistic regression analysis showed that advanced age [OR=1.146 (95%CI: 1.052-1.249)] and osteoporosis [OR=2.371 (95%CI: 1.042-5.396)] were the independent risk factors for MCI, while higher education [OR=0.073 (95%CI: 0.011-0.478)] was a protective factor. Age influenced all the aspects of MMSE scores (all P values <0.05). In addition, the analysis of the results suggested that subjects with regular folic acid supplementation got higher MMSE scores, especially in the aspect of language and praxis (P=0.002). On the contrary, patients with osteoporosis had lower attention and computing power scores (P=0.022). Conclusion The prevalence of MCI increased with age. Low education and osteoporosis may be the independent risk factors for MCI in the elderly population. Although no association was observed between folic acid supplementation and MCI, folic acid supplementation could improve the performance of language and praxis.