Jiawei Taohe Chengqi decoction inhibition of the notch signal pathway affects macrophage reprogramming to inhibit HSCs activation for the treatment of hepatic fibrosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiawei Taohe Chengqi Tang (JTCD) is a modified formulation of Traditional Chinese Medicine (TCM) known as Taohe Chengqi Decoction, which has been described in the ancient TCM literature "Treatise on Febrile Diseases". As a formula that can activate blood circulation and eliminate blood stasis and regulate Yin and Yang in traditional Chinese medicine applications, JTCD has been reported to be effective in the treatment of chronic liver disease and hepatic fibrosis (HF). AIM OF STUDY: The current study aimed to evaluate the effectiveness of JTCD in modulating hepatic macrophages by regulating the Notch signal pathway, and to further investigate the mechanisms underlying macrophage reprogramming that leads to HF. MATERIALS AND METHODS: Molecular assays were performed using in vitro cultures of human mononuclear THP-1 cells and human-derived hepatic stellate cells LX-2. CCl4-induced mice were utilized as an in vivo model to simulate HF. RESULTS: Our results demonstrated that JTCD exhibited dual effects by inhibiting hepatic stellate cell (HSCs) activation and modulating the polarisation of macrophages towards the M2 phenotype while decreasing the M1 phenotype. Network pharmacological analyses and molecular docking studies revealed that the Notch signal pathway was significantly enriched and played a crucial role in the therapeutic response of JTCD against HF. Moreover, through the establishment of a co-culture model, we validated that JTCD inhibited the Notch signal pathway in macrophages, leading to alterations in macrophage reprogramming, subsequent inhibition of HSC activation, and ultimately exerting anti-HF effects. CONCLUSION: In conclusion, our findings provide solid evidence for JTCD in treating HF, as it suppresses the Notch signal pathway in macrophages, regulates macrophage reprogramming, and inhibits HSC activation.

New Perspectives on Chinese Medicine in Treating Hepatic Fibrosis: Lipid Droplets in Hepatic Stellate Cells.

Hepatic fibrosis (HF) is a wound healing response featuring excessive deposition of the extracellular matrix (ECM) and activation of hepatic stellate cells (HSCs) that occurs during chronic liver injury. As an initial stage of various liver diseases, HF is a reversible pathological process that, if left unchecked, can escalate into cirrhosis, liver failure, and liver cancer. HF is a life-threatening disease presenting morbidity and mortality challenges to healthcare systems worldwide. There is no specific and effective anti-HF therapy, and the toxic side effects of the available drugs also impose a heavy financial burden on patients. Therefore, it is significant to study the pathogenesis of HF and explore effective prevention and treatment measures. Formerly called adipocytes, or fat storage cells, HSCs regulate liver growth, immunity, and inflammation, as well as energy and nutrient homeostasis. HSCs in a quiescent state do not proliferate and store abundant lipid droplets (LDs). Catabolism of LDs is characteristic of the activation of HSCs and morphological transdifferentiation of cells into contractile and proliferative myofibroblasts, resulting in the deposition of ECM and the development of HF. Recent studies have revealed that various Chinese medicines (e.g., Artemisia annua, turmeric, Scutellaria baicalensis Georgi, etc.) are able to effectively reduce the degradation of LDs in HSCs. Therefore, this study takes the modification of LDs in HSCs as an entry point to elaborate on the process of Chinese medicine intervening in the loss of LDs in HSCs and the mechanism of action for the treatment of HF.

Electron Conductive and Transparent Hydrogels for Recording Brain Neural Signals and Neuromodulation.

Recording brain neural signals and optogenetic neuromodulations open frontiers in decoding brain neural information and neurodegenerative disease therapeutics. Conventional implantable probes suffer from modulus mismatch with biological tissues and an irreconcilable tradeoff between transparency and electron conductivity. Herein, a strategy is proposed to address these tradeoffs, which generates conductive and transparent hydrogels with polypyrrole-decorated microgels as cross-linkers. The optical transparency of the electrodes can be attributed to the special structures that allow light waves to bypass the microgel particles and minimize their interaction. Demonstrated by probing the hippocampus of rat brains, the biomimetic electrode shows a prolonged capacity for simultaneous optogenetic neuromodulation and recording of brain neural signals. More importantly, an intriguing brain-machine interaction is realized, which involves signal input to the brain, brain neural signal generation, and controlling limb behaviors. This breakthrough work represents a significant scientific advancement toward decoding brain neural information and developing neurodegenerative disease therapies.

Mitochondrial-related microRNAs and their roles in cellular senescence.

Aging is a natural aspect of mammalian life. Although cellular mortality is inevitable, various diseases can hasten the aging process, resulting in abnormal or premature senescence. As cells age, they experience distinctive morphological and biochemical shifts, compromising their functions. Research has illuminated that cellular senescence coincides with significant alterations in the microRNA (miRNA) expression profile. Notably, a subset of aging-associated miRNAs, originally encoded by nuclear DNA, relocate to mitochondria, manifesting a mitochondria-specific presence. Additionally, mitochondria themselves house miRNAs encoded by mitochondrial DNA (mtDNA). These mitochondria-residing miRNAs, collectively referred to as mitochondrial miRNAs (mitomiRs), have been shown to influence mtDNA transcription and protein synthesis, thereby impacting mitochondrial functionality and cellular behavior. Recent studies suggest that mitomiRs serve as critical sensors for cellular senescence, exerting control over mitochondrial homeostasis and influencing metabolic reprogramming, redox equilibrium, apoptosis, mitophagy, and calcium homeostasis-all processes intimately connected to senescence. This review synthesizes current findings on mitomiRs, their mitochondrial targets, and functions, while also exploring their involvement in cellular aging. Our goal is to shed light on the potential molecular mechanisms by which mitomiRs contribute to the aging process.

Highly Stretchable Hydrogels as Wearable and Implantable Sensors for Recording Physiological and Brain Neural Signals.

Recording electrophysiological information such as brain neural signals is of great importance in health monitoring and disease diagnosis. However, foreign body response and performance loss over time are major challenges stemming from the chemomechanical mismatch between sensors and tissues. Herein, microgels are utilized as large crosslinking centers in hydrogel networks to modulate the tradeoff between modulus and fatigue resistance/stretchability for producing hydrogels that closely match chemomechanical properties of neural tissues. The hydrogels exhibit notably different characteristics compared to nanoparticles reinforced hydrogels. The hydrogels exhibit relatively low modulus, good stretchability, and outstanding fatigue resistance. It is demonstrated that the hydrogels are well suited for fashioning into wearable and implantable sensors that can obtain physiological pressure signals, record the local field potentials in rat brains, and transmit signals through the injured peripheral nerves of rats. The hydrogels exhibit good chemomechanical match to tissues, negligible foreign body response, and minimal signal attenuation over an extended time, and as such is successfully demonstrated for use as long-term implantable sensory devices. This work facilitates a deeper understanding of biohybrid interfaces, while also advancing the technical design concepts for implantable neural probes that efficiently obtain physiological information.

Epigenetic regulation in premature ovarian failure: A literature review.

Premature ovarian failure (POF), or premature ovarian insufficiency (POI), is a multifactorial and heterogeneous disease characterized by amenorrhea, decreased estrogen levels and increased female gonadotropin levels. The incidence of POF is increasing annually, and POF has become one of the main causes of infertility in women of childbearing age. The etiology and pathogenesis of POF are complex and have not yet been clearly elucidated. In addition to genetic factors, an increasing number of studies have revealed that epigenetic changes play an important role in the occurrence and development of POF. However, we found that very few papers have summarized epigenetic variations in POF, and a systematic analysis of this topic is therefore necessary. In this article, by reviewing and analyzing the most relevant literature in this research field, we expound on the relationship between DNA methylation, histone modification and non-coding RNA expression and the development of POF. We also analyzed how environmental factors affect POF through epigenetic modulation. Additionally, we discuss potential epigenetic biomarkers and epigenetic treatment targets for POF. We anticipate that our paper may provide new therapeutic clues for improving ovarian function and maintaining fertility in POF patients.

Research Progress on the Treatment of Premature Ovarian Failure Using Mesenchymal Stem Cells: A Literature Review.

Premature ovarian failure (POF) has become one of the main causes of infertility in women of childbearing age and the incidence of POF is increasing year by year, seriously affecting the physical and mental health of patients and increasing the economic burden on families and society as a whole. The etiology and pathogenesis of POF are complex and not very clear at present. Currently, hormone replacement therapy is mainly used to improve the symptoms of low estrogen, but cannot fundamentally solve the fertility problem. In recent years, stem cell (SC) transplantation has become one of the research hotspots in the treatment of POF. The results from animal experiments bring hope for the recovery of ovarian function and fertility in patients with POF. In this article, we searched the published literature between 2000 and 2020 from the PubMed database (https://pubmed.ncbi.nlm.nih.gov), and summarized the preclinical research data and possible therapeutic mechanism of mesenchymal stem cells (MSCs) in the treatment of POF. Our aim is to provide useful information for understanding POF and reference for follow-up research and treatment of POF.

Repair of a severe palm injury with anterolateral thigh and ilioinguinal flaps: A case report.

BACKGROUND: In daily life and work, there are more and more patients with trauma to the hand, which often results in skin and soft tissue defects. Although there are many repair methods, the function and appearance of the fingers will be adversely affected if the repair is inadequate. CASE SUMMARY: In the present report we describe an 18-year-old male patient whose right hand was mangled by a machine. X-ray imaging showed that a right hand bone (middle finger) was absent and the alignment was poor. After hospitalization, he was diagnosed with a severe right hand injury, skin and soft tissue defects, partial finger defects, and a skin degloving injury. He underwent reconstructive surgery with anterolateral thigh and ilioinguinal flaps. After two repair operations, satisfactory results were obtained, including good fracture healing, good skin flap shape, and good wrist joint function. CONCLUSION: This case highlights the good effect of anterolateral thigh and ilioinguinal flaps repair technique on severe palm injury.

Mitochondrial gene COX2 methylation and downregulation is a biomarker of aging in heart mesenchymal stem cells.

The mitochondria have been proven to be involved in processes of aging; however, the mechansims through which mitoepigenetics affect the cytological behaviors of cardiomyocytes during the aging process are not yet fully understood. In the present study, two senescence models were constructed, replicative senescence (RS) and stress­induced premature senescence (SIPS), using human heart mesenchymal stem cells (HMSCs). First, the differences in age­related gene expression levels and telomere length were compared between the HMSCs in the RS and SIPS models by PCR. Subsequently, protein expression and the mitochondrial DNA (mtDNA) methylation status of cytochrome c oxidase subunit II (COX2) was measured by western blot analysis and bisulfite genomic sequencing (BSP). Finally, the value of the DNA methyltransferase (Dnmt) inhibitor, 5­aza­2'­deoxycytidine (AdC), in delaying the senescence of HMSCs was evaluated. It was found that the p16, p27 and p53 mRNA expression levels increased in the senescent cells, whereas p21 mRNA expression did not. It was also found that telomere shortening only occurred in the RS model, but not in the SIPS model. Along with the senescence of HMSCs, COX2 gene methylation increased and its protein expression level significantly decreased. It was demonstrated that AdC inhibited COX2 methylation and downregulated COX2 expression. The addition of exogenous COX2 or the administration of AdC promoted cell proliferation and delayed cell aging. On the whole, the present study demonstrates that COX2 methylation and downregulation are biomarkers of HMSC senescence. Thus, COX2 may have potential for use as a therapeutic target of cardiovascular diseases and this warrants further investigation.

An elongated tract of polyQ in the carboxyl­terminus of human α1A calcium channel induces cell apoptosis by nuclear translocation.

An aberrant elongated tract of glutamine residues (polyQ) in proteins induces multiple diseases treated in the clinic. In our previous study of progressive myoclonic epilepsy (PME), using whole­exome sequencing, a mutant Cav2.1 protein with an aberrant elongated polyQ tract was identified in PME patients. To investigate the molecular mechanism and cell biology of this aberrant elongated polyQ tract, wild­type Cav2.1 with 13 polyQ repeats (Cav2.1 wt­Q13) and mutant­type Cav2.1 with 26 polyQ repeats (Cav2.1 mt­Q26) were prepared and introduced into human SH­SY5Y neuroblastoma cells. Using a WST­1 assay, it was revealed that Cav2.1 mt­Q26 markedly suppressed the proliferation of the SH­SY5Y cells, a result not observed for the Cav2.1 wt­Q13­transfected cells. It was also revealed that Cav2.1 mt and its truncated molecules suppressed cell proliferation by inducing apoptosis rather than arresting the cell cycle. Further investigations indicated a nuclear translocation phenomenon associated with the Cav2.1 mt molecules. Mechanistically, it was revealed that the Cav2.1 mt molecules activated the Bcl­2/Bax, caspase­3 and poly ADP­ribose polymerase (PARP) apoptotic pathways. The present study may provide new insights for interpreting the pathogenesis of PME and the relationship among polyQ, CACNA1A gene mutations and PME.

Long non­coding RNA GAS5 increases the radiosensitivity of A549 cells through interaction with the miR­21/PTEN/Akt axis.

Radioresistance hinders the therapeutic outcomes of radiotherapy in non­small cell lung cancer (NSCLC). Although long non­coding RNAs (lncRNAs) have been demonstrated to participate in the regulation of multiple cell behaviors, whether they can modulate the radiosensitivity of NSCLC and the underlying molecular mechanisms have not been well investigated. In the present study, it was revealed that NSCLC NCI­H460 cells were more sensitive to ionizing radiation (IR) than A549 cells. Using the RNA­Seq method, four highly differentially expressed lncRNAs were identified, including the growth arrest­specific transcript 5 (GAS5), syntaxin binding protein 5 antisense RNA 1 (STXBP5­AS1), metastasis associated lung adenocarcinoma transcript 1 (MALAT1) and X­inactive specific transcript (XIST), which were predicted to play roles in the acquisition of radiosensitivity. Using real­time quantitative PCR (qPCR), it was demonstrated that lncRNA GAS5 was significantly upregulated in NCI­H460 cells but not in A549 cells during IR. Mechanistically, it was demonstrated that overexpression of lncRNA GAS5 decreased the level of microRNA­21 (miR­21). Overexpression of lncRNA GAS5 or suppression of miR­21 markedly increased the IR­induced cell apoptosis of A549 cells. It was also demonstrated that overexpression of lncRNA GAS5 increased PTEN expression and suppressed Akt phosphorylation through the modulation of miR­21. Notably, it was revealed that IR enhanced the interaction between lncRNA GAS5 and the miR­21/PTEN/Akt axis. In summary, the present findings revealed that lncRNA GAS5 has a radiosensitization effect on NSCLC, indicating the potential application of lncRNA GAS5 in NSCLC radiotherapy.

Follicular helper T cell and memory B cell immunity in CHC patients.

Chronic hepatitis C (CHC) is associated with biological activity of T follicular helper (Tfh) cells and memory B cells (MBCs). However, the nature of Tfh cell subsets that are responsible for MBCs in CHC patients has not been evaluated. This study aimed to investigate Tfh and MBC immunity before and after direct-acting antiviral (DAA) therapy in patients with CHC. A total of 31 CHC patients and 15 healthy controls (HCs) were recruited. Individual patients were treated with sofosbuvir/ribavirin (SOF/RBV) or in combination with pegylated interferon alpha-2a (PEG-IFN-α-2a) for 12 weeks. Immunofluorescence revealed the frequency of ICOS+CD4+CXCR5+ active Tfh cells in liver tissue of CHC patients was higher than that of healthy control. Tfh and B cell co-culture experiments showed that Tfh2 cells from CHC patients have potential ability to induce B cell differentiation and IgG production. Flow cytometry showed that the frequencies of CD21-CD27+IgD- activated MBCs, ICOS+CD4+CXCR5+ activated Tfh cells, Tfh1 (IFN-γ+CD4+CXCR5+) cells, and Tfh2 (IL-4+CD4+CXCR5+) cells, but not of Tfh17 (IL-17+CD4+CXCR5+) cells, increased in CHC patients before and after DAA therapy. Collectively, ICOS+ Tfh, Tfh1, Tfh2 cells, and MBCs participated in the antiviral treatment process of SOF/RBV with or without PEG-IFN-α-2a in CHC patients, and their activity was further enhanced during the treatment. KEY MESSAGES: This study aimed to investigate Tfh cells and MBC immunity in CHC patients. CD21-CD27+IgD- activated MBCs increased in CHC patients before and after treatment. Tfh1 and Tfh2 cells increased in CHC patients before and after antiviral treatment. Intrahepatic activated Tfh cells increased in CHC patients before treatment. Tfh2 cells from CHC patients have a stronger ability to induce B cell differentiation.

GRIN3A and MAPT stimulate nerve overgrowth in macrodactyly.

As an uncommon and congenital condition, macrodactyly is characterized by an increase in the size of all the elements or structures of the digits or toes; however, the underlying pathogenesis remains to be fully elucidated. In the present study, the gene expression profiles of abnormal nerves were examined in three patients with macrodactyly using microarray analysis to identify potential genes contributing to nerve overgrowth. Gene expression profiling in the nerve tissue samples were scanned using the microarray and the differentially expressed genes were verified at the transcription level using reverse transcription­quantitative polymerase chain reaction analysis. Western blot analysis was used to determine the expression of target genes at the translational level. To confirm the upregulated genes during the process of nerve proliferation, SH­SY5Y cells were induced to differentiate into a neuronal­like phenotype using retinoic acid. A total of 165 genes showed significant changes (≥5­fold) in gene expression, which may be associated with the development of limbs in macrodactyly. Glutamate ionotropic receptor NMDA 3A (GRIN3A) and microtubule­associated protein tau (MAPT) were identified as important contributors in promoting nerve overgrowth. Furthermore, it was identified that GRIN3A and MAPT were regulated by the cAMP­protein kinase A and extracellular signal­regulated kinase 1/2 pathways, respectively. The identification of genes expressed at high levels in macrodactyly may reveal potential factors, which contribute to abnormal nerve proliferation and underpin the pathogenesis of macrodactyly, and provide potential application targets in nerve tissue regeneration engineering.

Interleukin­33­induced immune tolerance is associated with the imbalance of memory and naïve T­lymphocyte subsets.

The current study aimed to investigate the distribution of memory and naïve T cell (TN) subsets in hepatitis B virus (HBV)­infected patients at different immune stages and investigate the effect of interleukin 33 (IL­33) on the regulation of the T­cell subsets. The distributions of memory and naïve T cells were detected by flow cytometry. ELISA was conducted to assess the levels of IL­4, IL­5, IL­10, IL­12, interferon Î³ and tumor necrosis factor α. The expression levels of IL­33 and HBV x protein (HBx) were measured by reverse transcription­quantitative polymerase chain reaction and western blot analysis, respectively. By detecting TNs, central memory T cells (TCM) and effector memory T cells (TEM), it was identified that the proportions of TCM and TEM in CD4+ T cells were increased in patients with HBV. The trend observed for levels of CD8+ TCM and TEM was similar to that of CD4+ T cells in the immune tolerance and immune activation groups, however CD8+ TCM and TEM were significantly reduced in patients who underwent treatment. The CD8+ TEM cells appeared to be more sensitive to HBV activation and drug therapy. In addition, IL­33 stimulation was observed to induce imbalances of CD8+ TN and CD8+ TEM, and while the imbalances were directly regulated by HBx, IL­33 was not a key factor for the expression of HBx. CD8+ TEM cells may be a sensitive marker to assess the immune state of patients with HBV and the effect of clinical therapy. Treatment targeting IL­33 may be a potential method to reverse HBV­induced immune tolerance.

Posterior Thigh Flap Pedicled on the Cutaneous Vessels Arising From the Popliteo-posterior Intermediate Artery: A Report of 5 Cases.

Surgical repair of soft tissue defects of the knee and leg remains challenging. Using a case study approach, the anatomy of the popliteo-posterior intermediate cutaneous artery was examined, and a reverse island flap method was developed and implemented. After obtaining informed consent, 5 patients (1 woman, 4 men, age range 31 to 57 years) underwent the experimental use of a reverse island flap with a posterior thigh flap pedicled on the cutaneous vessels arising from the popliteo-posterior intermediate artery to repair soft-tissue defects of the knee and leg. The defects were caused by burned skin below the knee (n = 1), progressive skin necrosis in the knee after fracture surgery (n = 2), and skin infections associated with diabetes mellitus (n = 2). Skin defect sizes ranged from 15 cm x 5 cm to 30 cm x 12 cm. These large defects did not heal spontaneously; wound duration ranged from 1 week to 1 year, and all patients had refused defect repair with free flaps. Patients received posterior thigh flaps pedicled on the popliteo-posterior intermediate artery with areas ranging from 17 cm x 6 cm to 25 cm x 12 cm. All patients were treated with antibiotics and local dressings (iodoform and alcohol) changed daily post surgery, and blood supply was monitored by assessing the texture and color of the flap and venous regurgitation (ie, vein drainage disturbance). Four (4) of the five flaps survived completely. In 1 patient, partial survival of the flap, which had a good blood supply despite a venous circulation disorder, occurred: in this case, complete survival was achieved after treatment with a retrograde fascial flap and skin grafting. The appearance and texture of all flaps were satisfactory (ie, patients underwent only 1 operation, healing time was approximately 2 weeks, flap quality was close to normal skin, the donor site closed directly, and the shape and function of the knee and leg recovered well). No donor site abnormality was observed, and no postsurgical infection occurred. More research is needed, but the use of a reverse island flap with a posterior thigh flap pedicled on the cutaneous vessels arising from the popliteo-posterior intermediate artery may be a feasible option to repair soft tissue defects of the knee and leg.

A Lower Proportion of Regulatory B Cells in Patients with Henoch-Schoenlein Purpura Nephritis.

BACKGROUND: Henoch-Schoenlein purpura is the one of most common types of systemic vasculitis that involves impaired renal function and Henoch-Schoenlein purpura nephritis (HSPN). The diagnosis of this condition is largely based on immunohistologic detection of immunoglobulin A1-containing immune complex in the glomerular deposits of mesangium. Despite clinical advances, the etiopathogenesis of HSPN is still largely unknown. METHODS: In this study, we enrolled 25 newly diagnosed HSPN patients and 14 healthy controls. Then, fractions of B cell subtypes were determined in venous blood using flow cytometry. The serum interleukin (IL)-10 concentration was determined by enzyme-linked immunosorbent assay. RESULTS: Compared to those in healthy controls, the numbers of CD38+CD19+, CD86+CD19+, CD38+CD86+CD19+, and CD95+CD19+ B cells per microliter of blood were significantly higher in HSPN patients. In contrast, the numbers of CD5+CD19+, IL-10+CD19+, CD5+CD1d+CD19+, and IL-10+CD5+CD1d+CD19+ B cells per microliter of blood and the serum IL-10 concentration were significantly lower in HSPN patients. Following treatment, the numbers of CD38+CD19+ and CD86+CD19+ B cells per microliter of blood were significantly reduced in HSPN patients. However, the numbers of CD5+CD1d+CD19+, CD5+CD1d+IL-10+CD19+, and IL-10+CD19+ B cells per microliter of blood and the serum IL-10 concentration were significantly increased in HSPN patients following treatment. The estimated glomerular filtration rate (eGFR) was negatively correlated with the number of CD38+CD19+ B cells but positively correlated with the numbers of IL-10+CD19+, CD1d+CD5+CD19+, and IL-10+CD1d+CD5+CD19+B cells per microliter of blood and the serum IL-10 concentration. The 24-h urinary protein concentration was positively correlated with the number of CD38+CD19+B cells but negatively correlated with the numbers of IL-10+CD19+, CD1d+CD5+CD19+, and IL-10+CD1d+CD5+CD19+B cells per microliter of blood and the serum IL-10 concentration. CONCLUSION: Our results suggest that CD38+CD19+ and CD1d+CD5+CD19+ B cells (Bregs) contribute to the pathogenesis of HSPN.

A higher frequency of CD4(+)CXCR5(+) T follicular helper cells in patients with newly diagnosed Henoch-Schönlein purpura nephritis.

T follicular helper (TFH) cells play an important role in the humoral immune responses. The aim of this study was to examine the frequency of different subsets of CD4(+)CXCR5(+) TFH cells and B cells in patients with new-onset Henoch-Schönlein purpura nephritis (HSPN). The numbers of different subsets of CD4(+)CXCR5(+) TFH cells, B cells and the constituents of serum cytokines were detected in a total of 25 patients with newly diagnosed HSPN before and after treatment, and in 14 healthy controls (HC). The potential connection of these cells with the clinical characteristics in HSPN patients was analyzed. The numbers of circulating CD4(+)CXCR5(+), CD4(+)CXCR5(+)ICOS(+) and CD4(+)CXCR5(+)PD-1(+) TFH cells, CD86(+)CD19(+), CD38(+)CD19(+) B cells and serum IL-2, IL-4, IL-17A, IL-21 and IFN-γ were significantly higher in HSPN patients (p<0.05) than in HC. Before and after treatment the numbers of CD4(+)CXCR5(+) TFH cells were negatively correlated with the values of eGFR (r=-0.7162, p<0.05; r=-0.732, p<0.05, respectively). Similarly the numbers of CD4(+)CXCR5(+)PD-1(+) TFH cells were negatively correlated with 24-h urinary proteins (r=-0.4013, p<0.05; r=-0.7857, p<0.05, respectively), and the numbers of CD4(+)CXCR5(+)ICOS(+) TFH cells were positively correlated with the levels of serum IL-21 (r=0.5186, p<0.05; r=0.8503, p<0.05, respectively) and 24-h urinary protein (r=0.6045, p<0.05; r=0.833, p<0.05, respectively) in these patients, regardless of treatment. Following treatment the numbers of CD4(+)CXCR5(+), CD4(+)CXCR5(+)PD-1(+), and CD4(+)CXCR5(+)ICOS(+) TFH cells, as well as serum levels of IL-21 were significantly reduced, however IL-4 levels were noticeably increased (p<0.05). A higher frequency of circulating CD4(+)CXCR5(+) TFH cells existed in patients with HSPN and may be a viable therapeutic target.

Stimulus-dependent neuronal cell responses in SH-SY5Y neuroblastoma cells.

The aim of the present study was to elucidate the intracellular mechanisms that cause neuronal cell death following exposure to excitatory neurotransmitter­induced neurotoxicity, neurotoxins and oxidative stress. Human SH­SY5Y neuroblastoma cells were exposed to various stimuli, including glutamate, 6­hydroxydopamine (6­OHDA), and glucose oxidase, and cell viability was determined by MTT assay. Early apoptosis and necrosis were examined by Annexin V/propidium iodide double staining and flow cytometric analysis. Intracellular calcium ion concentration and mitochondrial membrane potential were assessed by Fluo­3a and JC­1 staining, respectively. In addition, protein expression of receptor­interacting protein (RIP) kinase 1 and RIP kinase 3 were evaluated by western blotting. Glutamate, 6­OHDA and glucose oxidase treatment decreased cell viability. Glutamate induced apoptosis and necrosis, whereas, 6­OHDA induced cell necrosis and glucose oxidase induced apoptosis. Furthermore, glutamate, 6­OHDA or glucose oxidase treatment significantly increased intracellular calcium concentrations (P<0.05). The effect of glutamate on mitochondrial membrane potential varied with high and low concentrations, whereas 6­OHDA and glucose oxidase significantly increased the mitochondrial membrane potential in the SH­SY5Y cells (P<0.05). Glutamate significantly upregulated expression levels of RIP kinase 1 (P<0.05), but not RIP kinase 3. These findings demonstrate that the response of SH­SY5Y cells varies with the stimuli. Furthermore, RIP kinase 1 may specifically regulate programmed necrosis in glutamate­mediated excitatory toxicity, but not in cell damage induced by either 6-OHDA or glucose oxidase.

The Effect of C-X-C Motif Chemokine 13 on Hepatocellular Carcinoma Associates with Wnt Signaling.

OBJECTS: To investigate the effect of CXCL13 (C-X-C motif chemokine 13) on hepatocellular carcinoma and clarify the potential mechanisms. METHODS: 32 patients with hepatocellular carcinoma and 12 healthy controls were recruited for analyzing the expression of CXCL13 by RT-PCR (reverse transcription-polymerase chain reaction). ELISA (enzyme-linked immune-sorbent assay) was used to test the concentration of serum CXCL13. The interaction between CXCL13 and Wnt signaling was analyzed by western blot. In vitro PBMCs cultured with HepG2 supernatant, the levels of IL-12, IL4, IL-6, and IL-17, and four IgG subclasses were detected by ELISA. RESULTS: The rate of high expression CXCL13 was 63.4% in advanced HCC patients, and the serum CXCL13 was also at a high level in stage IV HCC patients. Meanwhile CXCL13 level was positively correlated with serum ALT (Alanine Transaminase) and AST (Aspartate Aminotransferase). CXCL13 and Wnt/ß-catenin signaling shared a positive feedback loop. Furthermore, CXCL13 could obviously promote the expressions of IL-12 and IL-17, and induce IgG4 secreted by B cells. CONCLUSIONS: The effect of CXCL13 on promoting liver cancer is related to the activation of Wnt/ß-catenin pathway and the facilitation of IL-12, IL-17 and IgG4. CXCL13 plays an important role in the progression of HCC, and it may act as a potential target for the diagnosis and treatment of HCC.

Increased numbers of circulating ICOS⁺ follicular helper T and CD38⁺ plasma cells in patients with newly diagnosed primary biliary cirrhosis.

BACKGROUND: Aberrant activation of follicular helper T (TFH) and B cells is associated with the development of autoimmune diseases. However, little is known about the potential role of these cells in the development of primary biliary cirrhosis (PBC). AIM: This study aimed at characterizing the numbers of different subsets of circulating Tfh and B cells as well as evaluating their potential association with the levels of immunoglobulins and autoantibodies in newly diagnosed PBC patients. METHODS: The numbers of circulating CD27(+), CD38(+), CD86(+) and CD95(+) B cells as well as inducible T cell costimulator (ICOS)(+) and programmed death-1 (PD-1)(+), IL-21(+) TFH cells were examined in 58 patients with newly diagnosed PBC and 30 matched healthy controls (HCs). RESULTS: The numbers of circulating CD38(+)CD19(+), CD86(+)CD19(+), and CD95(+)CD19(+) B cells; CD3(+)CD4(+)CXCR5(+)ICOS(+) and CD3(+)CD4(+)CXCR5(+)PD-1(+) Tfh cells; and the levels of serum IL-21 in the PBC patients were significantly greater, but the numbers of CD27(+)CD19(+) B cells were significantly less than those in the HCs (p < 0.05). The numbers of CD3(+)CD4(+)CXCR5(+)ICOS(+) Tfh cells were positively correlated with the numbers of CD38(+)CD19(+) and CD86(+)CD38(+)CD19(+) B cells and the levels of serum anti-mitochondrial antibodies against M2 antigen (AMA-M2), AMA and immunolgubin M (IgM) in the PBC patients. The levels of serum IL-21 were positively correlated with the levels of serum AMA-M2, AMA, IgG and IgM, but negatively with the numbers of CD27(+)CD19(+) B cells in the PBC patients. CONCLUSIONS: Increased numbers of circulating ICOS(+) and IL-21(+) Tfh and CD38(+) plasma cells may be exhibited by patients with recent diagnoses of PBC.