Effect of Dendrobium nobile powder combined with conventional therapy on mild to moderate fatty liver

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)encompasses a variety of liver conditions impacting individuals who consume minimal or no alcohol.Recently,traditional Chinese medicine has been gradually used to treat mild to moderate fatty liver,among which Dendrobium nobile Lindl.powder has been affirmed by many doctors and patients to be effective.However,there is limited research on combining this treatment with standard therapies for mild to moderate NAFLD.AIM To survey the effect of combining Dendrobium nobile Lindl.powder with standard treatment on liver function and lipid metabolism disorder in patients with mild to moderate NAFLD.METHODS Eighty patients with mild to moderate NAFLD participated in this retrospective study,classified into two groups:The observation group(n=40)and the control group(n=40).In November 2020 and November 2022,the study was conducted at People’s Hospital of Chongqing Liang Jiang New Area.The control group received standard treatment,while the observation group received Dendrobium nobile Lindl.powder based on the control group.The study compared differences in traditional Chinese medicine clinical syndrome scores,liver fibrosis treatment,liver function indicators,lipid levels,and serum inflammatory factor levels before and after treatment,and we calculated the incidence of adverse reactions for both groups.RESULTS The total effective rate was 97.50%in the observation group and 72.5%in the control group.After 8 weeks of treatment,the main and secondary symptom scores remarkably decreased,especially in the observation group(P<0.05),and there was a significant reduction in the serum levels of hyaluronic acid(HA),laminin(LN),human rocollagen III(PC III),and collagen type IV(CIV).The levels of HA,LN,PC III,and CIV were significantly lower in the observation group(P<0.05).After 8 weeks,both groups indicated remarkable improvements in liver function and blood lipid levels,with the observation group having even lower levels(P<0.05).Serum levels of interleukin-1β,tumor necrosis factor-α,and interleukin-8 also dropped significantly.The observation group had a lower rate of adverse reactions(5.00%)compared to the control group(22.50%).CONCLUSION Adding Dendrobium nobile Lindl.powder to standard treatment has been found to remarkably improve symptoms and reduce inflammation in patients with mild to moderate fatty liver disease.It also enhances hepatic function and lipid profile,ameliorates liver fibrosis indices,and lowers the risk of side effects.Consequently,this therapeutic protocol shows promise for clinical implementation and dissemination.

Effect on Proliferation and Erythroid Differentiation of K562 Cells by IER3IP1-Knockdown

Objective： To investigate the effect on erythroid differentiation and proliferation of K562 cells by IER3IP1-knockdown with RNA interference targeting at IER3IP1 gene. Methods： The shRNA eukaryotic expression vectors targeting at IER3IP1 gene were designed and constructed. Inhibitory effect was detected by semiquantitative RT-PCR. The impacts on K562 cells by RNAi were studied by MTT assay, benzidine staining, light microscope and electron microscopy observation, cell cycles analysis, colony formation assay and RT-PCR. The expressions of erythroid differentiation correlated genes Gfi-lB, GPA and 7-globin were studied after being exposed to 0.2μmol/L imatinib for two days. Results： The shRNA eukaryotic expression vectors were successfully constructed. The expression of IER3IP1 gene was significantly inhibited with an inhibition efficiency of 76% （P〈0.01）. Compared with the control groups, bcr/abl mRNA level was increased in K562/shRNA-IER3IP1 group （P〈0.01）. The proliferation ability was enhanced （P〈0.01） and the proportion of cells at G0/G1 phase decreased but S phase increased （P〈0.05） in K562/shRNA-IER3IP1 group. Under electron microscopy, the amount of euchromatin increased but heterochromatin decreased. There were structural abnomalities in endocytoplasmic reticulum and clusters of vesicular. The percentage of benzidine staining positive cells and mRNA expression levels of Gfi-1B, GPA and γ-globin were all decreased after being exposed to 0.2 μmol/L STI571 for two days in K562/shRNA-IER3IP1 group （P〈0.01）. Conclusion： IER3IP1-knockdown can hinder the erythroid differentiation and elevate the proliferation level of K562 cells. IER3IP1 may play a role in erythroid differentiation and proliferation of K562 cells.

1~H NMR-based serum metabolic profiling in compensatedand decompensated cirrhosis

AIM: To study the metabolic profiling of serum samples from compensated and decompensated cirrhosis patients. METHODS: A pilot metabolic profiling study was conducted using three groups: compensated cirrhosis patients (n = 30), decompensated cirrhosis patients (n = 30) and healthy controls (n = 30). A 1H nuclear magnetic resonance (NMR)-based metabonomics approach was used to obtain the serum metabolic profiles of the samples. The acquired data were processed by multivariate principal component analysis and orthogonal partial least-squares discriminant analysis (OPLS-DA). RESULTS: The OPLS-DA model was capable of distinguishing between decompensated and compensated cirrhosis patients, with an R2Y of 0.784 and a Q2Y of 0.598. Twelve metabolites, such as pyruvate, phenylala-nine and succinate, were identified as the most influential factors for the difference between the two groups. The validation of the diagnosis prediction showed that the accuracy of the OPLS-DA model was 85% (17/20). CONCLUSION: 1H NMR spectra combined with pattern recognition analysis techniques offer a new way to diagnose compensated and decompensated cirrhosis in the future.

Expression of TRAIL in Mouse Uterine Endometrium during Embryo Implantation

Objective To investigate the expression of TRAIL in mouse uterine endometrium during embryo implantation and its role in the apoptosis of decidual cells. Methods Expression of TRAIL in uterine endometrium of pregnant mouse from d 1 to d 8 was detected with RT-PCR and immunohistochemistry. Results The expressed level of TRAIL mRNA in uterine endometrium of pregnant mouse from d 1 to d 8 was higher during embryo implantation than that prior to embryo implantation （P〈0.05）. No expression of TRAIL protein in mouse utrine endometrium was detected through d 1 to d 3. However, TRAIL protein was found in the luminal epithelial cells to which embryos attached on d 4. Moreover, TRAIL was expressed solely in decidual cells around invadting embryos through d 5 to d 6 while in trophoblastic cells adjacent to decidua through d 7 to d 8. Conclusion Apoptosis of luminal epithelial cells of endometrium induced by TRAIL could be one of mechanisms with which embryos penertrated the epithelial barrier, and apoptosis of both decidual cells and trophoblastic cells induced by TRAIL may play an important role during accruate invasion of trophoblastic cells.

Rapid construction of phosphatase and tensin homolog-deleted on chromosome ten gene recombinant adenovirus using the AdEasy system

Recent studies have shown that phosphatase and tensin homolog-deleted on chromosome ten （PTEN） gene plays an important role in ischemic brain damage and synaptic plasticity. The AdEasy system, which has been widely used, greatly simplifies preparation of recombinant adenovirus. Therefore, recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene （Ad-PTEN） was constructed using the AdEasy-1 system and was transfected into HEK293 cells for packaging and amplification. Infection efficiency and expression intensity were observed in primary cultured rat hippocampal neurons infected with Ad-PTEN in vitro. Results revealed a cytopathic effect in green fluorescent protein expression, which increased with prolonged time. After three cycles of amplification, the adenovirus titer was increased to an adequate titer for infecting hippocampal neurons. The entire process typically requires 4-5 weeks for completion. Results suggested that recombinant defective adenovirus vector carrying the PTEN gene was successfully and rapidly constructed using the AdEasy system.

Mitophagy deficiency activates stimulator of interferon genes activation and aggravates pathogenetic cardiac remodeling

Stimulator of interferon genes(STING)has recentlybeen found to play a crucial role in cardiac sterile inflammation and dysfunction.The role of stimulator of interferon genes(STING)in cardiac sterile inflammation and dysfunction has been recently discovered.This study aims to examine the involvement of STING in pathological cardiac remodeling and the mechanisms that govern the activation of the STING pathway.To investigate this,transverse aortic constriction(TAC)was performed on STING knockout mice to induce pressure over-load-induced cardiac remodeling.Subsequently,cardiac function,remodeling,and inflammation levels were evaluated.The STING pathway was found to be activated in the pressure overload-stressed heart and angiotensin II(Ang Il)-stimulated cardiac fibroblasts.Loss of STING expression led to a significant reduction in inflammatory responses,mitochondrial fragmenta-tion,and oxidative stress in the heart,resulting in attenuated cardiac remodeling and dysfunc-tion.Furthermore,the exacerbation of pressure overload-induced sTING-mediated inflammation and pathological cardiac remodeling was observed when mitophagy was sup-pressed through the silencing of Parkin,an E3 ubiquitin ligase.Taken together,these findings indicate that STING represents a newly identified and significant molecule implicated in the process of pathological cardiac remodeling and that mitophagy is an upstream mechanism that regulates STING activation.Targeting STING may therefore provide a novel therapeutic strategy for pathological cardiac remodeling and heart failure.

Tryptophan 2,3-dioxygenase-positive matrix fibroblasts fuel breast cancer lung metastasis via kynurenine-mediated ferroptosis resistance of metastatic cells and T cell dysfunction

Background:Tumor metastasis is a major threat to cancer patient survival.The organ-specific niche plays a pivotal role in tumor organotropic metas-tasis.Fibroblasts serve as a vital component of the metastatic microenviron-ment,but how heterogeneous metastasis-associated fibroblasts(MAFs)promote organotropic metastasis is poorly characterized.Here,we aimed to decipher the heterogeneity of MAFs and elucidate the distinct roles of these fibroblasts in pulmonary metastasis formation in breast cancer.Methods:Mouse models of breast cancer pulmonary metastasis were estab-lished using an in vivo selection method of repeated injections of metastatic cells purified from the mouse lung.Single-cell RNA-sequencing(scRNA-seq)was employed to investigate the heterogeneity of MAFs.Transgenic mice were used to examine the contribution of tryptophan 2,3-dioxygenase-positive matrix fibroblasts(TDO2^(+)MFs)in lung metastasis.Results:We uncovered 3 subtypes of MAFs in the lung metastatic microenviron-ment,and their transcriptome profiles changed dynamically as lung metastasis evolved.As the predominant subtype,MFs were exclusively marked by platelet-derived growth factor receptor alpha(PDGFRA)and mainly located on the edge of the metastasis,and T cells were enriched around MFs.Notably,high MF sig-natures were significantly associated with poor survival in breast cancer patients.Lung metastases were markedly diminished,and the suppression of T cells was dramatically attenuated in MF-depleted experimental metastatic mouse mod-els.We found that TDO2^(+)MFs controlled pulmonary metastasis by producing kynurenine(KYN),which upregulated ferritin heavy chain 1(FTH1)level in dis-seminated tumor cells(DTCs),enabling DTCs to resist ferroptosis.Moreover,TDO2^(+)MF-secreted chemokines C-C motif chemokine ligand 8(CCL8)and C-C motif chemokine ligand 11(CCL11)recruited T cells.TDO2^(+)MF-derived KYN induced T cell dysfunction.Conditional knockout of Tdo2 in MFs diminished lung metastasis and enhanced immune activation.Conclusions:Our study reveals crucial roles of TDO2^(+)MFs in promoting lung metastasis and DTCs’immune evasion in the metastatic niche.It suggests that targeting the metabolism of lung-specific stromal cells may be an effective treatment strategy for breast cancer patients with lung metastasis.

CGI-100 Specific shRNA Inhibits Proliferation and Induces Differentiation in Leukemia K562 Cells

Objective： To investigate the characteristics of CGI-100- knockdown K562 cells and the effect of CGI-100 RNA interference （RNAi） on matrine-treated K562 cells. Methods： Three oligonucleotides targeting CGI-100 gene and a pair of negative control containing the same nucleotide composition with a different sequence were devised and chemically synthesized. The inhibition efficiency of CGI-100 expression by shRNA-CGI-100 in K562 cells was determined using semiquantitative RT-PCR and dot blot hybridization. The effect of CGI-100 RNAi on the growth of K562 cells was examined using MTT assay and cell differentiation was measured by distinct approaches including flow cytometry, benzidine staining and electron microscope. After CGI-100-konckdown K562 cells were incubated with 0.2 mg/ml of matrine or 30 Ixmol/L of hemin for 48 h, the expression levels of Glycophorin A（GPA）（CD235a） and Growth factor independence-lB mRNA（Gfi-IB mRNA） were measured by RT-PCR and the protein levels of GPA, CD14 and CD15 were detected by flow cytometry. Results： The eukaryotic expression vectors of CGI-100 RNAi were successfully constructed. The K562/shRNA-CGI-100 cell line was established in which the inhibition efficiency of CGI-100 gene expression by shRNA-CGI-100 was 54%. CGI-100-knockdown inhibited the proliferation and induced erythroid differentiation in K562 cells. Compared with the control K562 cells, the K562/shRNA-CGI-100 cells showed decreased absorbance value detected by MTT assay, decreased enchromation, increased heterochromation, increased percentage of G0/Gx phase cells, decreased population of S phase cells, decreased PI （proliferation index of cells）, and elevated percentage of benzidine-positive cells. Moreover, the sensitivity of K562/shRNA-CGI-100 cells to either matrine or hemin was enhanced and the sensitivity of these cells to matrine was higher than that to hemin. Compared with the control K562 cells, matrine treatment in K562/shRNA-CGI-100 cells resulted in increased inhibitory rate of proliferation, elevated percentage of be nzidine-positive cells, obviously up-regulated mRNA expressions of GPA and Gfi-IB, and increased mean fluorescence intensity （MFI） of GPA. No CD14 expression was detected and no statistical significance was found for the detected CD15. Finally, the MFI of GPA increased in K562/shRNA-CGI-100 cells treated with hemin and was 1.7 times less than that in cells exposed to matrine. Conclusion： These results suggest that the function of CGI-100 gene is correlated with the deregulated proliferation and the block of erythroid differentiation in K562 cells and may also be involved in matrine-induced erythroid differentiation in K562 ceils.

Silk Fibroin-Based Hydrogel for Multifunctional Wearable Sensors

The flexible wearable sensors with excellent stretchability,high sensitivity and good biocompatibility are significantly required for continuously physical condition tracking in health management and rehabilitation monitoring.Herein,we present a high-performance wearable sensor.The sensor is prepared with nanocomposite hydrogel by using silk fibroin(SF),polyacrylamide(PAM),polydopamine(PDA)and graphene oxide(GO).It can be used to monitor body motions(including large-scale and small-scale motions)as well as human electrophysiological(ECG)signals with high sensitivity,wide sensing range,and fast response time.Therefore,the proposed sensor is promising in the fields of rehabilitation,motion monitoring and disease diagnosis.

Elevated neutrophil extracellular traps by HBV-mediated S100A9-TLR4/RAGE-ROS cascade facilitate the growth and metastasis of hepatocellular carcinoma

Background:Neutrophil extracellular traps(NETs)are considered significant contributors to cancer progression,especially metastasis.However,it is still unclear whether NETs are involved in hepatitis B virus(HBV)-related hepatocarcinogenesis and have potential clinical significance during evaluation and management for hepatocellular carcinoma(HCC).In this study,we aimed to investigate the functional mechanism of NETs in HBV-related hepatocarcinogenesis and their clinical significance.Methods:A total of 175 HCC patients with and without HBV infection and 58 healthy controls were enrolled in this study.NETs weremeasured in tissue specimens,freshly isolated neutrophils and blood serum from these patients,and the correlation of circulating serum NETs levels with malignancy was evaluated.The mechanism by which HBV modulates NETs formation was explored using cell-based studies.In addition,in vitro and in vivo experiments were further performed to clarify the functional mechanism of NETs on the growth and metastasis of HCC.Results:We observed an elevated level of NETs in blood serum and tissue specimens from HCC patients,especially those infected with HBV.NETs facilitated the growth and metastasis of HCC both in vitro and in vivo,which were mainly dominated by increased angiogenesis,epithelial-mesenchymal transition(EMT)-related cell migration,matrix metalloproteinases(MMPs)-induced extracellular matrix(ECM)degradation and NETs-mediated cell trapping.Inhibition of NETs generation by DNase 1 effectively abrogated the NETs-aroused HCC growth and metastasis.In addition,HBV-induced S100A9 accelerated the generation of NETs,which was mediated by activation of toll-like receptor(TLR4)/receptor for advanced glycation end products(RAGE)-reactive oxygen species(ROS)signaling.Further,circulatory NETs were found to correlate with viral load,TNM stage and metastasis status in HBV-related HCC,and the identified NETs could predict extrahepatic metastasis,with an area under the ROC curve(AUC)of 0.83 and 90.3%sensitivity and 62.8%specificity at a cutoff value of 0.32.Conclusions:Our findings indicated that activation of RAGE/TLR4-ROS signaling by HBV-induced S100A9 resulted in abundant NETs formation,which subsequently facilitated the growth and metastasis of HCC cells.More importantly,the identified circulatory NETs exhibited potential as an alternative biomarker for predicting extrahepatic metastasis in HBV-related HCC.

JAG1 enhances angiogenesis in triple-negative breast cancer through promoting the secretion of exosomal lncRNA MALAT1

Despite significant improvements in five-year survival rates due to early diagnosis and combination therapy, triple-negative breast cancer (TNBC) treatment remains a major challenge. Finding new and effective targets for diagnosis and drug therapy is urgent for TNBC patients. Jagged-1 (JAG1), one of the canonical ligands of the Notch signaling pathway, is involved in vascular budding and is a poor prognostic factor of TNBC. In this study, combined with quantitative real-time PCR, database analysis, animal experiments, and other means, JAG1 was confirmed to be related to the poor prognosis of TNBC patients. JAG1 was highly expressed in MDA-MB-231 Bone (231B) cells, with stronger invasion and metastasis ability than MDA-MB-231 (231) cells. Treatment of human vascular endothelial cells (HUVEC) with TNBC conditioned medium showed that TNBC JAG1 promoted the angiogenesis of HUVEC. Next, we detected the exosomes extracted from TNBC conditioned medium and found that JAG1 promoted the exosome secretion from 231 cells via ALIX-RAB11A/RAB35. In addition, we also found that the exosomes from JAG1 overexpressed TNBC cells contained more long non-coding RNA (lncRNA) MALAT1 , and MALAT1 promoted angiogenesis of HUVEC by targeting miR-140-5p . Finally, the angiogenesis-promoting effect of JAG1 in TNBC was further investigated by matrix gel assay. In conclusion, we reveal that JAG1 has a pro-invasion effect on TNBC and is involved in microenvironment angiogenesis by promoting exosome secretion and the MALAT1-miR-140-5p-JAG1/VEGFA pathway.

Wnt3a enhances bone morphogenetic protein 9-induced osteogenic differentiation of C3H10T1/2 cells

Background Bone morphogenetic protein 9 （BMP9） and Wnt/13-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells （MSCs）, but the role of Wnt/13-catenin signaling pathway in BMP9-induced osteogenic differentiation is not well understood. Thus, our experiment was undertaken to investigate the interaction between BMP9 and Wnt/13-catenin pathway in inducing osteogenic differentiation of MSCs. Methods C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP9, Wnt3a, and BMPg＋Wnt3a. ALP, the early osteogenic marker, was detected by quantitative and staining assay. Later osteogenic marker, mineral calcium deposition, was determined by Alizarin Red S staining. The expression of osteopotin （OPN）, osteocalcin （OC）, and Runx2 was analyzed by Real time PCR and Western blotting. In vivo animal experiment was carried out to further confirm the role of Wnt3a in ectopic bone formation induced by BMP9. Results The results showed that Wnt3a enhanced the ALP activity induced by BMP9 and increased the expressions of OC and OPN, with increase of mineral calcium deposition in vitro and ectopic bone formation in vivo. Furthermore, we also found that Wnt3a increased the level of Runx2, an important nuclear transcription factor of BMP9. Conclusion Canonical Wnt/13-catenin signal pathway may play an important role in BMP9-induced osteogenic differentiation of MSCs, and Runx2 may be a linkage between the two signal pathways.

Overexpression of Jak1 Activating Mutants in Hepatocytes Is Insufficient to Generate Hepatocellular Carcinoma in Zebrafish

Hepatocellular carcinoma （HCC）, a major subtype of liver cancers, is a prevalent human malignancy worldwide. In men, HCC is the fifth frequently diagnosed cancer but the second most common cause of cancer death. In women, it ranks seventh in cancer diagnosis and sixth in cancer-related death （Jemal et al., 2011）. Unlike some other cancers, such as breast cancer and colon cancer, the molecular etiology of HCC re- mains largely unknown. Infection of hepatitis virus is considered as a major risk factor in the development of liver cancers （Parkin, 2006）. Currently, there are limited options to treat HCC except for chemotherapy. Elucidating molecular mechanism of hepatocyte transformation will help develop new treatments for cancer. The widely accepted multi-step progression of carcinogenesis consists of genetic alterations which regulate cell proliferation, apoptosis and so on （Vogelstein and Kinzler, 1993）. Moreover, abnormal activation of signaling pathways has been proposed as an oncogenic driver for cancer development. For example, Kras activation occurs in 7% of human liver cancer patients. Activated Kras is sufficient to induce robust liver tumorigenesis in transgenic animal models （Nguyen et al., 2011）.

Comparative proteomic analysis of plasma from bipolar depression and depressive disorder:identification of proteins associated with immune regulatory

Bipolar disorder(BD)is a debilitating psychiatric mood dis-order affecting approximately 1%-3%of the population worldwide(Merikangas,2007).Bipolar disorder is characterized by recurrent episodes of depression,hypomania,mania,or mixed states,and it has a poor outcome,with high rates of relapse,lingering residual symptoms,cognitive impairment,and functional impairment(Moreno et al.,2007)Although various etiopathological hypotheses concerning the disease have been reported,the pathophysiology underlying BD remains poody understood(Gawryluk and Young,2011).

Artificial intelligence reinforced upconversion nanoparticle-based lateral flow assay via transfer learning

The combination of upconverting nanoparticles(UCNPs)and immunochromatography has become a widely used and promising new detection technique for point-of-care testing(POCT).However,their low luminescence efficiency,non-specific adsorption,and image noise have always limited their progress toward practical applications.Recently,artificial intelligence(AI)has demonstrated powerful representational learning and generalization capabilities in computer vision.We report for the first time a combination of AI and upconversion nanoparticle-based lateral flow assays(UCNP-LFAs)for the quantitative detection of commercial internet of things(IoT)devices.This universal UCNPs quantitative detection strategy combines high accuracy,sensitivity,and applicability in the field detection environment.By using transfer learning to train AI models in a small self-built database,we not only significantly improved the accuracy and robustness of quantitative detection,but also efficiently solved the actual problems of data scarcity and low computing power of POCT equipment.Then,the trained AI model was deployed in IoT devices,whereby the detection process does not require detailed data preprocessing to achieve real-time inference of quantitative results.We validated the quantitative detection of two detectors using eight transfer learning models on a small dataset.The AI quickly provided ultra-high accuracy prediction results(some models could reach 100%accuracy)even when strong noise was added.Simultaneously,the high flexibility of this strategy promises to be a general quantitative detection method for optical biosensors.We believe that this strategy and device have a scientific significance in revolutionizing the existing POCT technology landscape and providing excellent commercial value in the in vitro diagnostics(IVD)industry.

Extracellular vesicles:Emerging tools as therapeutic agent carriers

Extracellular vesicles(EVs)are secreted by both eukaryotes and prokaryotes,and are present in all biological fluids of vertebrates,where they transfer DNA,RNA,proteins,lipids,and metabolites from donor to recipient cells in cell-to-cell communication.Some EV components can also indicate the type and biological status of their parent cells and serve as diagnostic targets for liquid biopsy.EVs can also natively carry or be modified to contain therapeutic agents(e.g.,nucleic acids,proteins,polysaccharides,and small molecules)by physical,chemical,or bioengineering strategies.Due to their excellent biocompatibility and stability,EVs are ideal nanocarriers for bioactive ingredients to induce signal transduction,immunoregulation,or other therapeutic effects,which can be targeted to specific cell types.Herein,we review EV classification,intercellular communication,isolation,and characterization strategies as they apply to EV therapeutics.This review focuses on recent advances in EV applications as therapeutic carriers from in vitro research towards in vivo animal models and early clinical applications,using representative examples in the fields of cancer chemotherapeutic drug,cancer vaccine,infectious disease vaccines,regenerative medicine and gene therapy.Finally,we discuss current challenges for EV therapeutics and their future development.