Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is in-volved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregu-lation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatic-ally reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of hu-man STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Fur-thermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regula-ting the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a po-tential inhibitory factor affecting the occurrence and progression of STAD.

This study aimed to explore the therapeutic mechanisms of Colquhounia Root Tablets(CRT) in treating diabetic kidney disease(DKD) by integrating biomolecular network mining with animal model verification. By analyzing clinical transcriptomics data, an interaction network was constructed between candidate targets of CRT and DKD-related genes. Based on the topological eigenvalues of network nodes, 101 core network targets of CRT against DKD were identified. These targets were found to be closely related to multiple pathways associated with type 2 diabetes, immune response, and metabolic reprogramming. Given that immune-inflammatory imbalance driven by metabolic reprogramming is one of the key pathogenic mechanisms of DKD, and that many core network targets of CRT are involved in this pathological process, receptor for advanced glycation end products(RAGE)-reactive oxygen species(ROS)-phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)-nuclear factor-κB(NF-κB)-NOD-like receptor family pyrin domain containing 3(NLRP3) signaling axis was selected as a candidate target for in-depth research. Further, a rat model of DKD induced by a high-sugar, high-fat diet and streptozotocin was established to evaluate the pharmacological effects of CRT and verify the expression of related targets. The experimental results showed that CRT could effectively correct metabolic disturbances in DKD, restore immune-inflammatory balance, and improve renal function and its pathological changes by inhibiting the activation of the RAGE-ROS-PI3K-AKT-NF-κB-NLRP3 signaling axis. In conclusion, this study reveals that CRT alleviates the progression of DKD through dual regulation of metabolic reprogramming and immune-inflammatory responses, providing strong experimental evidence for its clinical application in DKD.
Animals ; Diabetic Nephropathies/metabolism* ; Receptor for Advanced Glycation End Products/genetics* ; NF-kappa B/genetics* ; Signal Transduction/drug effects* ; Rats ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Proto-Oncogene Proteins c-akt/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Phosphatidylinositol 3-Kinases/genetics* ; Reactive Oxygen Species/metabolism* ; Humans ; Plant Roots/chemistry* ; Rats, Sprague-Dawley ; Tablets/administration & dosage*

Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is in-volved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregu-lation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatic-ally reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of hu-man STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Fur-thermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regula-ting the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a po-tential inhibitory factor affecting the occurrence and progression of STAD.

Multidrug resistance (MDR) is the main cause of clinical treatment failure and poor prognosis in cancer. Targeting P-glycoprotein (P-gp) has been regarded as an effective strategy to overcome MDR. In this work, we reported our preclinical studies of the triazolo[1,5-a]pyrimidine-based compound WS-716 as a highly potent, specific, and orally active P-gp inhibitor. Through direct binding to P-gp, WS-716 inhibited efflux function of P-gp and specifically reversed P-gp-mediated MDR to paclitaxel (PTX) in multiple resistant cell lines, without changing its expression or subcellular localization. WS-716 and PTX synergistically inhibited formation of colony and 3D spheroid, induced apoptosis and cell cycle arrest at G2/M phase in resistant SW620/Ad300 cells. In addition, WS-716 displayed minimal effect on the drug-metabolizing enzyme cytochrome P4503A4 (CYP3A4). Importantly, WS-716 increased sensitivity of both pre-clinically and clinically derived MDR tumors to PTX in vivo with the T/C value of 29.7% in patient-derived xenograft (PDX) models. Relative to PTX treatment alone, combination of WS-716 and PTX caused no obvious adverse reactions. Taken together, our preclinical studies revealed therapeutic promise of WS-716 against MDR cancer, the promising data warrant its further development for cancer therapy.

OBJECTIVE: To observe the clinical therapeutic effect of filiform-fire needling of "Biaoben acupoint combination" on the sequelae of patients with coronavirus disease 2019 (COVID-19) during the recovery period. METHODS: A total of 33 patients with COVID-19 during the recovery period were treated with filiform-fire needling at the acupoints of Mingmen (GV 4), Shenzhu (GV 12), Gaohuang (BL 43), Zusanli (ST 36) and Shangjuxu (ST 37), etc., once every other day, 3 times a week, and 3 times was one course of treatment and totally 2 courses of treatment were required. The TCM symptom, Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD) scores, pulmonary function indexes (forced vital capacity [FVC], forced expiratory volume in one second [FEV1], peak expiratory flow [PEF]) and chest CT imaging change were observed before and after treatment, and the therapeutic effect was evaluated. RESULTS: After treatment, the scores of TCM symptom, HAMA and HAMD were decreased compared with those before treatment (P<0.05), and the levels of FVC, FEV1 and PEF were increased compared with those before treatment (P<0.05), and the recovery rate of 22 patients with pulmonary ventilation dysfunction was 86.4% (19/22). After treatment, the lung shadow area was smaller than that before treatment (P<0.05). The effective rate of 25 patients with lung CT abnormalities was 84.0% (21/25). After treatment, 23 cases were cured, 5 cases were markedly effective, 4 cases were effective, 1 case was ineffective, the cured and markedly effective rate was 84.8%. CONCLUSION The filiform-fire needling of "Biaoben acupoint combination" could significantly reduce the sequelae of cough, fatigue, chest tightness, etc. and mental symptoms such as anxiety and depression in patients with COVID-19 during the recovery period, and promote inflammatory exudation absorption of pulmonary lesion and improve lung ventilation function.
Acupuncture Points ; Acupuncture Therapy ; COVID-19/therapy* ; Humans ; Lung ; Vascular Surgical Procedures

OBJECTIVE: To investigate effects of physiological hypoxic conditions on suspension and adherence of embryoid bodies (EBs) during differentiation of human induced pluripotent stem cells (hiPSCs) and explore the underlying mechanisms. METHODS: EBs in suspension culture were divided into normoxic (21% O₂) and hypoxic (5% O₂) groups, and those in adherent culture were divided into normoxic, hypoxic and hypoxia + HIF-1α inhibitor (echinomycin) groups. After characterization of the pluripotency with immunofluorescence assay, the hiPSCs were digested and suspended under normoxic and hypoxic conditions for 5 days, and the formation and morphological changes of the EBs were observed microscopically; the expressions of the markers genes of the 3 germ layers in the EBs were detected. The EBs were then inoculated into petri dishes for further culture in normoxic and hypoxic conditions for another 2 days, after which the adhesion and peripheral expansion rate of the adherent EBs were observed; the changes in the expressions of HIF-1α, β-catenin and VEGFA were detected in response to hypoxic culture and echinomycin treatment. RESULTS: The EBs cultured in normoxic and hypoxic conditions were all capable of differentiation into the 3 germ layers. The EBs cultured in hypoxic conditions showed reduced apoptotic debris around them with earlier appearance of cystic EBs and more uniform sizes as compared with those in normoxic culture. Hypoxic culture induced more adherent EBs than normoxic culture (P < 0.05) with also a greater outgrowth rate of the adherent EBs (P < 0.05). The EBs in hypoxic culture showed significantly up-regulated mRNA expressions of β-catenin and VEGFA (P < 0.05) and protein expressions of HIF-1 α, β-catenin and VEGFA (P < 0.05), and their protein expresisons levels were significantly lowered after treatment with echinomycin (P < 0.05). CONCLUSION Hypoxia can promote the formation and maturation of suspended EBs and enhance their adherence and post-adherent proliferation without affecting their pluripotency for differentiation into all the 3 germ layers. Our results provide preliminary evidence that activation of HIF-1α/β-catenin/VEGFA signaling pathway can enhance the differentiation potential of hiPSCs.
Echinomycin/metabolism* ; Embryoid Bodies/metabolism* ; Humans ; Hypoxia/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; beta Catenin/metabolism*

Objective:To explore the therapeutic effect of Fuzheng Huayu capsule on nonalcoholic fatty liver fibrosis induced by high trans fatty acid and high sugar diet in mice. Method:Forty SPF male C57/B6 mice were randomly divided into 4 groups （normal group， model group， Obelcholinic acid group， and Fuzheng Huayu capsule group）， with 10 mice in each group. Except 10 mice in the normal group， nonalcoholic fatty liver fibrosis was induced by high-fat and high-sugar diet for 24 weeks in the other 30 mice. From the 25th week of modeling， 4 groups received intragastric administration for 4 weeks， once a day： Fuzheng Huayu capsule group 4.8 g·kg^-1. Oxycholic acid group 10 mg·kg^-1. Model control group and normal group received the same volume of normal saline. Liver tissue and serum samples were collected at the end of the 28th week. The pathological changes of liver tissue of mice in each group were observed by hematoxylin-eosin （HE） staining， the degree of liver fibrosis was observed by Sirius red staining， the degree of lipid deposition was observed by oil red O staining， the content of hydroxypropylamine （Hyp） in liver tissue was determined by alkaline hydrolysis， and the change of triglyceride （TG） in liver tissue was detected by enzyme method. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST） activities and fasting blood glucose （FBG） were detected by kit method， enzyme-linked immunosorbent assay （ELISA） was used to detect fasting Insulin （INS） level and calculate the changes of insulin resistance index （HOMA-IR）， liver fibrosis related mRNA and proteins of were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， Western blot and Immunohistochemistry. Result:Compared with the normal group， the liver tissue in the model group showed significant collagen fiber deposition， at mostly F2-F3 fibrosis stages， with increased number of inflammatory foci in liver tissue， obvious balloon degeneration and fatty degeneration of liver cells， significantly increased contents of Hyp and TG in liver tissue （P<0.01）， significantly increased activities of ALT and AST in serum and levels of FBG， INS and HOMA-IR （P<0.01）， and increased type I collagen （Col-Ⅰ）， Col-Ⅳ， α-smooth muscle agonist protein （α-SMA） and transforming growth factor-β （TGF-β） protein and mRNA in liver tissue. Compared with the model group， the collagen fiber deposition in liver tissue of mice in Fuzheng Huayu capsule group was significantly reduced， at mostly F0-F1 fibrosis stages， with significantly improved balloon-like change of hepatocytes， and significantly reduced number of inflammatory foci in lobules （P<0.05，P<0.01）. Compared with the model group， Fuzheng Huayu capsule can significantly reduce the content of Hyp in liver tissue， the levels of serum ALT and AST， and the expression of Col-Ⅰ， Col-Ⅳ， α-SMA and TGF-β genes and proteins in mice （P<0.05，P<0.01）. Conclusion:Fuzheng Huayu capsule has a good therapeutic effect on nonalcoholic fatty liver fibrosis induced by high trans fatty acid and high sugar diet in mice.

Aged ; Cognitive Dysfunction/epidemiology* ; Cross-Sectional Studies ; Geriatric Assessment ; Humans ; Independent Living ; Sarcopenia/epidemiology*

OBJECTIVETo explore the clinical effect of low dose pituitrin in children with septic shock.

METHODSA total of 48 pediatric cases with septic shock, in whom 6 hours, conventional treatment could not reverse shock from January 2008 to December 2010, were selected for this study. The patients were divided into two groups randomly (completely random design) (control group 24, remedial group 24). The conventional treatment included antibiotics/fluid resuscitation/correcting acid-base imbalance, glucocorticoid, organ (heart/lung) support, dopamine 1 - 15 µg/(kg·min) and norepinephrine 0.5 - 2 µg/(kg·min) pumped in continuously in the control group. In initial 6 hours the same treatment was given to the remedial group, while low dose pituitrin (0.01 - 0.03 U/min) was pumped additionally during the rest of time. The therapeutic effect on correcting shock was evaluated in both groups.

RESULTSThe total effective rate was 76.2% in the remedial group and 40.0% in the control group; the mortality was 33.3% and 60% respectively. The difference between both groups was significant (P = 0.025).

CONCLUSIONLow dose pituitrin could improve the clinical effect significantly in children with septic shock in whom 6 hours conventional treatment failed to correct shock, shorten the total periods of treatment, and decrease mortality.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Norepinephrine ; therapeutic use ; Pituitary Hormones, Posterior ; administration & dosage ; therapeutic use ; Shock, Septic ; drug therapy ; Treatment Outcome ; Vasoconstrictor Agents ; therapeutic use

Objective To investigate the effects of duration of untreated psychiatry (DUP) on the white matter integrity in first-episode medication-free patients with schizophrenia. Methods The Chinese version of Nottingham Onset Schedule was used to assess the DUP of 39 first-episode medication-free patients with schizophrenia. According to the median of DUP, the 39 patients were grouped into long-DUP group and short-DUP group. Diffusion weighted images of the 39 patients' whole brains were acquired with a Half-Fourier Acquired Single-Shot Turbo Spin Echo (HASTE) sequence.After being preprocessed with DTI-studio and statistical parametric mapping software (SPM5), the fractional anisotropy (FA) images of the 2 groups were compared by two-sample t-test with SPM5 software. The differences of gender, age, education level and total scores of Positive and Negative Syndrome Scale (PANSS) scores between the 2 groups were also detected. Results No significant difference was noted on gender, age, education level, PANSS scores between the 2 groups (P＞0.05).Subjects of long-DUP group showed significantly reduced FA value in the right anterior cingulate fasciculus (x=8, y=40, z=24) and left prefrontal white matter thresholded (x=32, y=34, z=4) as compared with that of short-DUP group at a level of P＜0.001 (uncorrected). Conclusion Extension of the duration of DUP will reduce the white matter integrity in first-episode medication-free patients with schizophrenia.