Immunogenic Cell Death-related Signature Evaluates the Tumor Microenvironment and Predicts the Prognosis in Diffuse Large B-Cell Lymphoma.

Current literatures suggest a growing body of evidence highlighting the pivotal role of Immunogenic Cell Death (ICD) in multiple tumor types. Nevertheless, the potential and mechanisms of ICD in diffuse large B-cell lymphoma (DLBCL) remain inadequately studied. To address this gap, our current study aims to examine the impact of ICD on DLBCL and identify a corresponding gene signature in DLBC. Using the expression profiles of ICD-associated genes, the gene expression omnibus (GEO) samples were segregated into ICD-high and ICD-low subtypes utilizing non-negative matrix factorization clustering. Next, univariate and LASSO Cox regression analyses were employed to establish the ICD-related gene signature. Subsequently, the CIBERSORT tool, ssGSEA, and ESTIMATE algorithm were utilized to examine the association between the signature and tumor immune microenvironment of DLBC. Finally, the oncoPredict algorithm was implemented to evaluate the drug sensitivity prediction of DLBCL patients. These findings suggest that the immune microenvironment of the ICD-high group with a poor prognosis was significantly suppressed. An 8-gene ICD-related signature was identified and validated to prognosticate and evaluate the tumor immune microenvironment in DLBCL. Similarly, the high-risk group exhibited a worse prognosis compared to the low-risk group, and the immune function was considerably suppressed. Moreover, the results of oncoPredict algorithm indicated that patients in the high-risk group exhibited higher sensitivity to Cisplatin, Cytarabine, Epirubicin, Oxaliplatin, and Vincristine with low IC50. In conclusion, the present study provides novel insights into the role of ICD in DLBCL by identifying a new biomarker for the disease and may have implications for the development of immune-targeted therapies for the tumor.

Circular Intronic RNA circTTN Inhibits Host Gene Transcription and Myogenesis by Recruiting PURB Proteins to form Heterotypic Complexes.

Muscle cell growth plays an important role in skeletal muscle development. Circular RNAs (circRNAs) have been proven to be involved in the regulation of skeletal muscle growth and development. In this study, we explored the effect of circTTN on myoblast growth and its possible molecular mechanism. Using C2C12 cells as a functional model, the authenticity of circTTN was confirmed by RNase R digestion and Sanger sequencing. Previous functional studies have showed that the overexpression of circTTN inhibits myoblast proliferation and differentiation. Mechanistically, circTTN recruits the PURB protein on the Titin (TTN) promoter to inhibit the expression of the TTN gene. Moreover, PURB inhibits myoblast proliferation and differentiation, which is consistent with circTTN function. In summary, our results indicate that circTTN inhibits the transcription and myogenesis of the host gene TTN by recruiting PURB proteins to form heterotypic complexes. This work may act as a reference for further research on the role of circRNA in skeletal muscle growth and development.

Tunable moiré bands and strong correlations in small-twist-angle bilayer graphene.

According to electronic structure theory, bilayer graphene is expected to have anomalous electronic properties when it has long-period moiré patterns produced by small misalignments between its individual layer honeycomb lattices. We have realized bilayer graphene moiré crystals with accurately controlled twist angles smaller than 1° and studied their properties using scanning probe microscopy and electron transport. We observe conductivity minima at charge neutrality, satellite gaps that appear at anomalous carrier densities for twist angles smaller than 1°, and tunneling densities-of-states that are strongly dependent on carrier density. These features are robust up to large transverse electric fields. In perpendicular magnetic fields, we observe the emergence of a Hofstadter butterfly in the energy spectrum, with fourfold degenerate Landau levels, and broken symmetry quantum Hall states at filling factors ±1, 2, 3. These observations demonstrate that at small twist angles, the electronic properties of bilayer graphene moiré crystals are strongly altered by electron-electron interactions.

Topologically Protected Helical States in Minimally Twisted Bilayer Graphene.

In minimally twisted bilayer graphene, a moiré pattern consisting of AB and BA stacking regions separated by domain walls forms. These domain walls are predicted to support counterpropogating topologically protected helical (TPH) edge states when the AB and BA regions are gapped. We fabricate designer moiré crystals with wavelengths longer than 50 nm and demonstrate the emergence of TPH states on the domain wall network by scanning tunneling spectroscopy measurements. We observe a double-line profile of the TPH states on the domain walls, only occurring when the AB and BA regions are gapped. Our results demonstrate a practical and flexible method for TPH state network construction.

van der Waals Heterostructures with High Accuracy Rotational Alignment.

We describe the realization of van der Waals (vdW) heterostructures with accurate rotational alignment of individual layer crystal axes. We illustrate the approach by demonstrating a Bernal-stacked bilayer graphene formed using successive transfers of monolayer graphene flakes. The Raman spectra of this artificial bilayer graphene possess a wide 2D band, which is best fit by four Lorentzians, consistent with Bernal stacking. Scanning tunneling microscopy reveals no moiré pattern on the artificial bilayer graphene, and tunneling spectroscopy as a function of gate voltage reveals a constant density of states, also in agreement with Bernal stacking. In addition, electron transport probed in dual-gated samples reveals a band gap opening as a function of transverse electric field. To illustrate the applicability of this technique to realize vdW heterostructuctures in which the functionality is critically dependent on rotational alignment, we demonstrate resonant tunneling double bilayer graphene heterostructures separated by hexagonal boron-nitride dielectric.

Intrinsic disorder in graphene on transition metal dichalcogenide heterostructures.

Semiconducting transition metal dichalchogenides (TMDs) are a family of van der Waals bonded materials that have recently received interest as alternative substrates to hexagonal boron nitride (hBN) for graphene, as well as for components in novel graphene-based device heterostructures. We elucidate the local structural and electronic properties of graphene on TMD heterostructures through scanning tunneling microscopy and spectroscopy measurements. We find that crystalline defects intrinsic to TMDs induce substantial electronic scattering and charge carrier density fluctuations in the graphene. These signatures of local disorder explain the significant degradation of graphene device mobilities using TMD substrates, particularly compared to similar graphene on hBN devices.

Clinical Study on Low-Frequency Electrical Pulse Acupoint Stimulation Combined with Pelvic Floor Muscle Exercise in the Treatment of Urinary Incontinence after Radical Prostatectomy.

BACKGROUND: Urinary incontinence (UI) is a common complication after radical prostatectomy (RP). It has a great influence on the postoperative quality of life of patients. This study aims to explore the clinical efficacy of low-frequency electrical pulse acupoint stimulation combined with pelvic floor muscle exercise in the treatment of UI after RP. METHODS: The clinical data of 129 patients with UI after receiving RP in our hospital from July 2020 to July 2023 were retrospectively analysed. A total of 65 patients who received pelvic floor muscle exercise from July 2020 to January 2022 were set as the reference group. Of these patients, four were excluded, resulting in the inclusion of 61 cases. A total of 64 patients who received low-frequency electrical pulse acupoint stimulation combined with pelvic floor muscle exercise from February 2022 to July 2023 were classified into the observation group. Of these patients, four were excluded, and 60 cases were finally included. SPSS 23.0 was used to analyse the use of urine pads, recovery time of urinary control and improvement of urination in the two groups. RESULTS: Before treatment, no significant difference existed in the use of urine pads, urination condition, maximum flow rate, maximum cystometric capacity, maximum urethral closure pressure, abdominal leak point pressure and scores on Short-Form-36 Health Survey (SF-36) in both groups (p > 0.05). After treatment, the observation group had significantly lower use of urinary pads, urination frequency and leakage times; Significantly shorter recovery time of urinary control (p < 0.05); And significantly higher maximum flow rate, maximum cystometric capacity, maximum urethral closure pressure, abdominal leak point pressure and SF-36 scores than the reference group (p < 0.05). CONCLUSIONS: The combination of low-frequency electrical pulse acupoint stimulation and pelvic floor muscle exercise can improve clinical symptoms, shorten the recovery time of urinary control and improve urodynamics and quality of life in patients with UI after RP.

Biomarkers and coptis chinensis activity for rituximab-resistant diffuse large B-cell lymphoma: Combination of bioinformatics analysis, network pharmacology and molecular docking.

BACKGROUND: Rituximab resistance is one of the great challenges in the treatment of diffuse large B-cell lymphoma (DLBCL), but relevant biomarkers and signalling pathways remain to be identified. Coptis chinensis and its active ingredients have antitumour effects; thus, the potential bioactive compounds and mechanisms through which Coptis chinensis acts against rituximab-resistant DLBCL are worth exploring. OBJECTIVE: To elucidate the core genes involved in rituximab-resistant DLBCL and the potential therapeutic targets of candidate monomers of Coptis chinensis. METHODS: Using the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP), the Similarity Ensemble Approach and Swiss Target Prediction, the main ingredients and pharmacological targets of Coptis chinensis were identified through database searches. Through the overlap between the pharmacological targets of Coptis chinensis and the core targets of rituximab-resistant DLBCL, we identified the targets of Coptis chinensis against rituximab-resistant DLBCL and constructed an active compound-target interaction network. The targets and their corresponding active ingredients of Coptis chinensis against rituximab-resistant DLBCL were molecularly docked. RESULTS: Berberine, quercetin, epiberberine and palmatine, the active components of Coptis chinensis, have great potential for improving rituximab-resistant DLBCL via PIK3CG. CONCLUSION: This study revealed biomarkers and Coptis chinensis-associated molecular functions for rituximab-resistant DLBCL.

Electrical Acupoint Stimulation with Low-Frequency Pulse in the Treatment of Urinary Incontinence after Prostatectomy.

OBJECTIVE: This study aimed to explore the clinical effect of electrical acupoint stimulation with low-frequency pulse in the treatment of urinary incontinence after prostatectomy. METHODS: This study selected 104 patients who underwent radical prostatectomy in Pujiang People Hospital from April 2019 to April 2022 as the research subjects, and they were divided into the study group (SG, n = 51, electrical acupoint stimulation with low-frequency pulse) and the control group (CG, n = 53, traditional pelvic floor muscle exercise) in accordance with the therapeutic regimen. In addition, clinical and follow-up data were analysed, and the number of urine pads used before and after treatment, recovery time of urinary continence, scores of 36-Item Short-form Health Survey (SF-36), clinical curative efficacy and incidence of adverse reactions in both groups were compared. RESULTS: Before treatment, no remarkable difference in the number of urine pads used was observed between the two groups (p > 0.05). After treatment, the number of urine pads used in the two groups was less than that before treatment, and the number of urine pads used in the SG was less than that in the CG (p < 0.001). The SG had overtly shorter recovery time of urinary continence, higher scores in eight dimensions of SF-36 and higher treatment efficiency than the CG (all p < 0.05), with no remarkable difference in the incidence of adverse reactions in both groups (p > 0.05). CONCLUSIONS: Electrical acupoint stimulation with low-frequency pulse, as a safe and ideal treatment, can shorten the recovery time of postoperative urinary continence ability, reduce the incidence of urinary incontinence and improve the quality of life of patients.

Circulating inflammatory cytokines in relation to the risk of renal cell carcinoma: A gender-specific two-sample Mendelian randomization study.

BACKGROUND: Currently there is no specific molecular biomarker for the diagnosis and treatment of renal cell carcinoma (RCC). Here we performed a gender-specific two-sample Mendelian randomization analysis to systematically assess the effects of circulating cytokines on RCC. METHODS: We have employed cis-quantitative trait loci as instrumental variables for the protein levels and expression of circulating cytokines. We estimated the causal effects of circulating cytokines on RCC risk in males and females with several Mendelian randomization methods. RESULTS: We observed a significant causal effect of Eotaxin on the increased risk of RCC in males (Odds ratio [OR] = 2.546, 95% confidence interval [CI] = 1.617-4.010, p value = 5.496 × 10-5), but not in females (OR = 1.352, 95% CI = 0.766-2.388, p value = 0.298). Besides, we also identified several cytokines as potentially associated with RCC in males including RANTES, MCP3, PDGFbb, TRAIL, and several other cytokines as potentially associated with RCC in females including sICAM and SCGFb. CONCLUSION: Our study highlighted that a higher level of circulating Eotaxin is causally associated with an increased risk of RCC in males but not in females. Further studies are needed to elucidate the exact mechanism and its potential application in the prognosis and treatment of RCC.

An HPK1 inhibitor enhanced the tumour response to anti-PD-1 immunotherapy in non-Hodgkin's lymphoma.

Anti-PD-1 immunotherapy has been widely applied in patients with some types of lymphoma. Classical Hodgkin's lymphoma (cHL) is highly sensitive to immunotherapy, but non-Hodgkin's lymphoma (NHL) does not show a good response. Studies have indicated that haematopoietic progenitor kinase 1 (HPK1) suppresses T cells and reduces antitumour immunity. Therefore, HPK1 inhibitors may restore and elicit antitumour immune responses and are promising candidate drug targets for cancer immunotherapy. We first explored the Gene Expression Profile Interactive Analysis (GEPIA) database and predicted that HPK1 expression was increased in diffuse large B-cell lymphoma (DLBCL) and associated with Nod-like receptor protein 3 (NLRP3) expression. We investigated whether an HPK1 inhibitor could enhance the tumour response to anti-PD-1 immunotherapy in NHL and the association between HPK1 and NLRP3 expression. Employing shHPK1 and an inhibitor, we demonstrated that the HPK1 inhibitor increased anti-PD-1-mediated T-cell cytotoxicity in BJAB and WSU-DLCL2 cells cocultured with peripheral blood mononuclear cells (PBMCs). HPK1 inhibitor treatment increased PD-1, PD-L1, Bax, p53 and NK-kB expression but decreased NLRP3 expression, indicating that the HPK1 inhibitor promoted apoptosis and blocked the NLRP3 inflammasome pathway to affect anti-PD-1-mediated T-cell cytotoxicity. Moreover, the HPK1 inhibitor enhanced the efficiency of anti-PD-1 immunotherapy in vivo in a zebrafish xenograft model of NHL. In summary, this study provides evidence that an HPK1 inhibitor enhanced the tumour response to anti-PD-1 immunotherapy in NHL by promoting apoptosis and blocking the NLRP3 pathway. These findings provide a potential therapeutic option for NHL combining HPK1 inhibitor treatment and anti-PD-1 immunotherapy.

Advances of research of Fc-fusion protein that activate NK cells for tumor immunotherapy.

The rapid development of bioengineering technology has introduced Fc-fusion proteins, representing a novel kind of recombinant protein, as promising biopharmaceutical products in tumor therapy. Numerous related anti-tumor Fc-fusion proteins have been investigated and are in different stages of development. Fc-fusion proteins are constructed by fusing the Fc-region of the antibody with functional proteins or peptides. They retain the bioactivity of the latter and partial properties of the former. This structural and functional advantage makes Fc-fusion proteins an effective tool in tumor immunotherapy, especially for the recruitment and activation of natural killer (NK) cells, which play a critical role in tumor immunotherapy. Even though tumor cells have developed mechanisms to circumvent the cytotoxic effect of NK cells or induce defective NK cells, Fc-fusion proteins have been proven to effectively activate NK cells to kill tumor cells in different ways, such as antibody-dependent cell-mediated cytotoxicity (ADCC), activate NK cells in different ways in order to promote killing of tumor cells. In this review, we focus on NK cell-based immunity for cancers and current research progress of the Fc-fusion proteins for anti-tumor therapy by activating NK cells.

New semen freezing method for chicken and drake using dimethylacetamide as the cryoprotectant.

The aim is to optimize the dimethylacetamide (DMA) straw freezing technology of Black silkies rooster semen through the handy patent equipment, screening the formula of freezing basic extender and optimizing the DMA addition method, and then by comparing the fertility of DMA straw frozen semen with the pellet frozen semen. After the DMA straw freezing technology is optimized, it is extended to the Youxian Partridge drake semen. The result showed that the frozen sperm motility of Lake and Ravie (LR) group is 64%, the fertility 49.57% and the hatchability 91.52%, all of which are superior to those of FEB, Beltsville Poultry Semen Extender (BPSE) and Lake (P < 0.05). The sperm motility of adding DMA stock solution is 59%, which is superior to adding DMA directly into diluted semen (P > 0.05). The fertility and hatchability of DMA straw group are 77.61% and 92.30%, respectively, and it is significantly higher than those in the pellet group (P < 0.01; P < 0.05). The fresh drake sperm motility of induction collection method is 71%, the massage collection method 61% and the frozen drake sperm motility of induction 33% while the massage 19%. The fertility of frozen drake semen group is 85.93%, while that of the fresh semen group is 88.17%. The frozen drake semen fertility of the highest batch is 93.8%. In conclusion, the world's advanced fertility of frozen semen can be obtained both in the chicken and drake through the optimized DMA straw freezing technology and the method of screening freeze-resistant individuals.

[Cloning and bioinformatics analysis of SLA-DR genes in Hunan Shaziling pigs].

In order to clone class II DRA and DRB genes of swine leukocyte antigen (SLA) in Hunan Shaziling pigs, to analyze their characteristics and polymorphism and to provide immunological basic parameters for xenotransplantation from pigs to humans. SLA-DRA and SLA-DRB genes in two Shaziling pigs with the absence of porcine endogenous retrovirus (PERV) env-c were amplified by RT-PCR, cloned into PUCm-T vectors, sequenced and analyzed through BLAST in NCBI and related software in ExPASY. The obtained SLA-DRA and SLA-DRB genes of Shaziling pigs were 1,177 and 909 nucleotides in length with their accession numbers in Genbank as EF143987 and EF143988. Bioinformatics analyses have shown that they both contain opening reading frame (ORF) and encode 252 and 266 amino acids respectively. Comparing the ORF and protein sequences of the Shaziling SLA-DRA and SLA-DRB genes with their counterpart sequences of human, the homologies of nucleotide sequences were 83% and 83%, and the homologies of amino acid sequences 83 % and 79% respectively. Further comparison with SLA sequences published in GenBank indicated that SLA-DRB gene found in Shaziling pigs has polymorphism while the homology of SLA-DRA gene is up to 100 % .

Evolution of the electronic band structure of twisted bilayer graphene upon doping.

The electronic band structure of twisted bilayer graphene develops van Hove singularities whose energy depends on the twist angle between the two layers. Using Raman spectroscopy, we monitor the evolution of the electronic band structure upon doping using the G peak area which is enhanced when the laser photon energy is resonant with the energy separation of the van Hove singularities. Upon charge doping, the Raman G peak area initially increases for twist angles larger than a critical angle and decreases for smaller angles. To explain this behavior with twist angle, the energy separation of the van Hove singularities must decrease with increasing charge density demonstrating the ability to modify the electronic and optical properties of twisted bilayer graphene with doping.

[Research of porcine endogenous retrovirus in Shaziling pigs].

To provide basic parameters of evaluating the biological safety of xenotransplantation from pig to human, ear tissues from 31 individuals were randomly collected from a Shazi Ling pig population. PCR and RT-PCR were performed to detect porcine endogenous retrovirus (PERV) proviral DNA and mRNA respectively. The sensitivity of the PCR was evaluated using a positive control. To study tissue distribution, RT-PCR of pol, gag and env was performed in the kidney, heart, liver, lung and spleen of 3 individuals. Finally, env-A, env-B and env-C were amplified, sequenced and analyzed using the BLAST software in NCBI (National Center for Biotechnology Information). The results showed PERV proviral DNA and mRNA could be detected in all 31 individuals by PCR and RT-PCR, respectively. env-A, env-B and env-C were only detected in 2 individuals, while in the other 29 individuals, only env-A and env-B but not env-C was detected. The quantity of DNA in PCR amplification of PERV genes should be more than 15 ng. RT-PCR results showed gag, pol, env-A and env-B were expressed in the kidney, heart, liver, lung and spleen of all 3 individuals, but env-C was not. Sequencing of env genes in Shazi Ling pigs revealed that while there was no difference in env-A sequence when compared to other herd in GenBank, there were 2 and 10 bp differences in the sequences of env-B and env-C respectively, suggesting that env gene is polymorphic in different pig strains. PERV exists in the Shazi Ling pig population and the predominant subtype is PERV-A, B. The distribution of PERV displays no significant tissue specificity and env-C is absent in 93.5% (29/31) of the individuals. The results indicate that Shazi Ling pig may have great potential value as a candidate in xenotransplantation.

[Analysis of effect of microsatellite markers on Escherichia coli F4 (K88) receptor in different swine breeds].

Four microsatellite markers linked with Escherichia-coli F4 receptor gene were selected from chromosome 13 of pig (authorization) to analyze difference in hereditary between the genotype and F4 receptor adhesive appearance. Results showed that the microsatellite markers were highly polymorphic and the heterozygosities ranged from 0.6117 to 0.7500. The value of PIC was over 0.5749 and the heredity diversity of Chinese pig breeds more higher than oversea's. On F4ab adhesive appearance of S0222, the difference among genotypes was significant in SZL breed. The AC genotype of SW458 had no adhesive appearance in SZL and LW breeds. Marker SW458 can thus be potentially used as a genetic marker for E. coli F4 resistive gene.

[Porcine endogenous retrovirus in Daweizi pigs in Hunan].

OBJECTIVE: To detect porcine endogenous retrovirus (PERV) in Daweizi pigs and to provide basic parameters of evaluating the biological safety for xenotransplantation from pigs to humans. METHODS: Ear tissues from 42 individuals were randomly collected from a Daweizi pig population. PCR and RT-PCR were performed to detect PERV proviral DNA and mRNA respectively. Finally, env-A, env-B, and env-C were amplified, sequenced, and analyzed using the BLAST software in National Center for Biotechnology Information. RESULTS: PERV proviral DNA and mRNA could be detected in the 42 individuals by PCR and RT-PCR, respectively. env-A, env-B and env-C were detected in all the individuals. Compared with other pig species (AY288779, DQ011794 and AY534304), there was 1 and 8 bp differences in the sequences of env-A and env-C, while no difference in env-B. CONCLUSION: PERV exists and has transcriptive activity in Daweizi pigs. The predominate subtype is PERV-ABC. Env genes are firstly cloned and sequenced in Daweizi pigs and there are polymorphism in the breed. As to the biological safety, the breed was not suitable as a donor in xenotransplantation.

Polymorphism of E. coli F18 receptor gene in different pig breeds.

The genetic variation of E. coli F18 receptor(ECF18R) loci in 867 pigs including Large Yorkshire, Landrace, Duroc, Ningxiang pig, Shaziling pig and Daweizi pig was detected by PCR-RFLP with Hin6 I. The results showed as follows: The three exotic breeds including Large Yorkshire, Landrace and Duroc were proved to be polymorphic at the Hin6 I-RFLP site. The average susceptibility genotype frequency of AG and GG in the exotic breeds was 0.94, G allelic frequency was 0.76. The C-->A mutation frequency at M307 site was very low in exotic breeds. The resistance genotype frequency of AA was 0.06. But the three native breeds including Ningxiang pig, Shaziling pig and Daweizi pig presented only GG genotype and no variation at the Hin6 I-RFLP site. The chi 2 value of ECF18R genotype distribution at the Hin6I-RFLP site in different pigs indicated that ECF18R genotype distribution of every exotic pig breed had significant or highly significant differences with three native pig breeds'. The ECF18R genotype distribution of three native pig breeds was the same completely. The others had no difference except that Landrace ECF18R genotype distribution had significant differences with Duroc's in the exotic breeds of pig.