Integrating network pharmacology with pharmacological research to elucidate the mechanism of modified Gegen Qinlian Decoction in treating porcine epidemic diarrhea.

Porcine Epidemic Diarrhea Virus (PEDV) poses a significant threat to neonatal piglets, particularly due to the limited efficacy of existing vaccines and the scarcity of efficacious therapeutic drugs. Gegen Qinlian Decoction (GQD) has been employed for over two millennia in treating infectious diarrhea. Nonetheless, further scrutiny is required to improve the drug's efficacy and elucidate its underlying mechanisms of action. In this study, a modified GQD (MGQD) was developed and demonstrated its capacity to inhibit the replication of PEDV. Animal trials indicated that MGQD effectively alleviated pathological damage in immune tissues and modulated T-lymphocyte subsets. The integration of network analysis with UHPLC-MS/MS facilitated the identification of active ingredients within MGQD and elucidated the molecular mechanisms underlying its therapeutic effects against PEDV infections. In vitro studies revealed that MGQD significantly impeded PEDV proliferation in IPEC-J2 cells, promoting cellular growth via virucidal activity, inhibition of viral attachment, and disruption of viral biosynthesis. Furthermore, MGQD treatment led to increased expression levels of IFN-α, IFN-ß, and IFN-λ3, while concurrently decreasing the expression of TNF-α, thereby enhancing resistance to PEDV infection in IPEC-J2 cells. In conclusion, our findings suggest that MGQD holds promise as a novel antiviral agent for the treatment of PEDV infections.

A preoperative parathyroid scan is important for the total removal of the transplanted parathyroid tissue in recurrent secondary hyperthyroidism: A case report and literature review.

RATIONALE: Secondary hyperparathyroidism was one of mineral and bone disorders owing to chronic kidney disease. Patients who suffer from secondary hyperparathyroidism would receive medical treatment or parathyroidectomy with or without autotransplantation (AT). However, some patients receiving parathyroidectomy with AT have recurrent hyperparathyroidism, which impacts their lives. Patients with recurrent hyperparathyroidism may present persistent hypercalcemia and hyperphosphatemia, which would cause cardiovascular disease, like atherosclerosis. PATIENT CONCERNS: A 63-year-old female of Asian descent with chronic kidney disease who suffered from recurrent hyperparathyroidism for twice. The patient underwent parathyroidectomy with AT in the left thigh when secondary hyperparathyroidism happened. After 3 months, recurrent hyperparathyroidism happened. DIAGNOSIS: The patient was diagnosed with recurrent hyperparathyroidism due to chronic kidney disease with hyperparathyroidism status post parathyroidectomy with AT in the left thigh. Our patient also suffered from mineral and bone disorder. INTERVENTION: Two parathyroid adenoma in the left thigh were found. However, one of them was too small to found in the operation. Therefore, autograftectomy of the large one was performed. However, hyperparathyroidism happened again. This time, the autograftectomy was performed under dual phase Tc-99m MIBI (99m Tc-methoxy isobutyl isonitrile) parathyroid scintigraphy and it succeeded. OUTCOMES: After secondary autograftectomy, the value of intact parathyroid hormone was surveyed immediately and dropped by two-third followed by gradual reduction in the following weeks. The calcemia and phosphatemia were back to normal gradually. LESSONS: In our case, importance of scintigraphy in the parathyroidectomy was confirmed.

αPD-1-mesoCAR-T cells partially inhibit the growth of advanced/refractory ovarian cancer in a patient along with daily apatinib.

Epithelial ovarian cancer (EOC) is the leading cause of death among gynecological malignancies in China. In particular, advanced/refractory ovarian cancer lacks effective targeted therapies due to the immunosuppressive and proangiogenic tumor microenvironment. Mesothelin (MSLN) has been found to be highly expressive in most EOC. Targeting MSLN by antibodies or chimeric antigen receptor-modified T (CAR-T) cells and immune checkpoint blockades as well as apatinib, an anti-angiogenic drug, have been used in patients with refractory ovarian cancer. Apatinib was reported to promote the infiltration of CD8+ T cells in lung cancer. However, the combination therapy of CAR-T secreting anti-PD-1 antibody with apatinib in EOC has not been reported. CASE PRESENTATION: Here we report a case of refractory EOC in a patient who had relapsed after multiline chemotherapy. The patient received autologous T cells that contained sequences encoding single-chain variable fragments specific for MSLN and full-length antibody for PD-1 (αPD-1). The modified T cells were called αPD-1-mesoCAR-T cells. After infusion, the copy number and PD-1 antibody secretion of the CAR-T cells were increased in the blood. By application of multimodality tumor tracking, MRI of the liver showed shrinkage of metastatic nodules from average diameter of 71.3-39.1 mm at month 2. The patient achieved partial response and survived more than 17 months. IL-6 levels in the patient fluctuated from the baseline to 2-4-folds after treatment, but side effects were mild with only grade 1 hypertension and fatigue. CONCLUSION: αPD-1-mesoCAR-T cell therapy combined with apatinib demonstrates a potential therapeutic effect on advanced refractory ovarian cancer. TRIAL REGISTRATION NUMBER: NCT03615313.

Mapping Cortical Integration of Sensory and Affective Pain Pathways.

Pain is an integrated sensory and affective experience. Cortical mechanisms of sensory and affective integration, however, remain poorly defined. Here, we investigate the projection from the primary somatosensory cortex (S1), which encodes the sensory pain information, to the anterior cingulate cortex (ACC), a key area for processing pain affect, in freely behaving rats. By using a combination of optogenetics, in vivo electrophysiology, and machine learning analysis, we find that a subset of neurons in the ACC receives S1 inputs, and activation of the S1 axon terminals increases the response to noxious stimuli in ACC neurons. Chronic pain enhances this cortico-cortical connection, as manifested by an increased number of ACC neurons that respond to S1 inputs and the magnified contribution of these neurons to the nociceptive response in the ACC. Furthermore, modulation of this S1→ACC projection regulates aversive responses to pain. Our results thus define a cortical circuit that plays a potentially important role in integrating sensory and affective pain signals.

Granger causality analysis of rat cortical functional connectivity in pain.

OBJECTIVE: The primary somatosensory cortex (S1) and the anterior cingulate cortex (ACC) are two of the most important cortical brain regions encoding the sensory-discriminative and affective-emotional aspects of pain, respectively. However, the functional connectivity of these two areas during pain processing remains unclear. Developing methods to dissect the functional connectivity and directed information flow between cortical pain circuits can reveal insight into neural mechanisms of pain perception. APPROACH: We recorded multichannel local field potentials (LFPs) from the S1 and ACC in freely behaving rats under various conditions of pain stimulus (thermal versus mechanical) and pain state (naive versus chronic pain). We applied Granger causality (GC) analysis to the LFP recordings and inferred frequency-dependent GC statistics between the S1 and ACC. MAIN RESULTS: We found an increased information flow during noxious pain stimulus presentation in both S1[Formula: see text]ACC and ACC[Formula: see text]S1 directions, especially at theta and gamma frequency bands. Similar results were found for thermal and mechanical pain stimuli. The chronic pain state shares common observations, except for further elevated GC measures especially in the gamma band. Furthermore, time-varying GC analysis revealed a negative correlation between the direction-specific and frequency-dependent GC and animal's paw withdrawal latency. In addition, we used computer simulations to investigate the impact of model mismatch, noise, missing variables, and common input on the conditional GC estimate. We also compared the GC results with the transfer entropy (TE) estimates. SIGNIFICANCE: Our results reveal functional connectivity and directed information flow between the S1 and ACC during various pain conditions. The dynamic GC analysis support the hypothesis of cortico-cortical information loop in pain perception, consistent with the computational predictive coding paradigm.

Tracking Changes in Brain Network Connectivity under Transcranial Current Stimulation.

Noninvasive transcranial brain stimulation has been widely used in experimental and clinical applications to perturb the brain activity, aiming at promoting synaptic plasticity or enhancing functional connectivity within targeted brain regions. However, there are different types of neurostimulations and various choices of stimulation parameters; how these choices influence the intermediate neurophysiological effects and brain connectivity remain incompletely understood. We propose several quantitative methods to investigate the brain connectivity of an epileptic patient before and after transcranial alternating/direct current stimulation (tACS/tDCS). The neuro-feedback derived from our analyses may provide useful cues for the effectiveness of neurostimulation.

MicroRNA­363­3p inhibits hepatocarcinogenesis by targeting HMGA2 and is associated with liver cancer stage.

The importance of microRNAs (miRNAs) in cancer development has been widely recognized in recent decades. In the present study, the function and mechanism of miRNA­363­3p (miR­363­3p), formerly characterized as a tumor suppressor, in the hepatocarcinogenesis of liver cancer cells was investigated. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was applied to detect the expression of miR­363­3p in liver cancer tissues. Cell proliferation, survival and migration capacities were determined by MTT, colony formation and wound­healing assays, respectively. The targeting of high mobility group AT­hook 2 (HMGA2) mRNA by miR­363­3p was confirmed by bioinformatics analysis, and RT­qPCR, luciferase reporter and western blot assays. The correlation between the expression levels of HMGA2 and miR­363­3p was analyzed. The RT­qPCR results revealed that the levels of miR­363­3p were downregulated in liver cancer tissues. Cellular assays validated that miR­363­3p exerted tumor suppressing functions, including the inhibition of cell proliferation, survival and migration abilities in two liver cancer cell lines. Bioinformatics prediction and subsequent experiments demonstrated that HMGA2 was a direct target of miR­363­3p. Restoration of the expression of HMGA2 in miR­363­3p mimic­transfected cells reversed the tumor suppressing effects caused by miR­363­3p. Finally, there was a significant negative correlation between the expression levels of HMGA2 and miR­363­3p in liver cancer tissues. miR­363­3p was identified as an important tumor suppressor in liver cancer via targeting HMGA2, which may have potential benefits in liver cancer therapy.

miR-145-5p Increases Osteoclast Numbers In Vitro and Aggravates Bone Erosion in Collagen-Induced Arthritis by Targeting Osteoprotegerin.

BACKGROUND Osteoprotegerin (OPG) inhibits bone resorption and binds with strong affinity to receptor activator of NF κB ligand (RANKL), thereby preventing RANKL from binding to its receptor RANK. Osteoclasts have documented effects on bone erosion of rheumatoid arthritis (RA). The aim of this study was to examine the role of miR-145-5p in the regulation of RA osteoclast differentiation and bone erosion. MATERIAL AND METHODS Expression of microRNA-145-5p in human peripheral blood mononuclear cells (PBMC) and synovial tissue was assayed by real-time polymerase chain reaction (RT-PCR). OPG, RANK, and RANKL expression in RAW-264.7 cells was examined by RT-PCR and Western blot analysis. Osteoclast formation was detected by tartrate-resistant acid phosphatase (TRAP) staining. The effect of miR-145-5p on predicted target mRNAs was examined by luciferase reporter assays. Collagen-induced arthritis (CIA) was induced by injecting DBA/1 mice with bovine type II collagen (CII), and miR-145-5p agomir was administered by intravenous injection. Morphological changes in the CIA joint were assessed by micro-computed tomography (CT) and histopathology. RESULTS miR-145-5p levels significantly increased in RA PBMC and synovial tissue compared with normal PBMC and osteoarthritis (OA) tissue. After transfection of RAW-264.7 cells with miR-145-5p, RANK and RANKL expression increased significantly, while OPG expression decreased significantly. TRAP staining results showed osteoclast numbers increased. Micro-CT analysis of the arthritic joints showed that the miR-145-5p agomir caused bone erosion in mice, and histopathological analysis revealed that miR-145-5p agomir aggravates cartilage erosion. CONCLUSIONS Our findings indicate that administration of miR-145-5p aggravates joint erosion in CIA mice. This suggests that miR-145-5p is a potential target for the treatment of RA.

[Correlation between the genetic polymorphisms of apolipoprotein M with the susceptibility to rheumatic diseases of Chinese Han populastion in Lanzhou].

Objective To investigate the relationship between the genetic polymorphisms of apolipoprotein M (ApoM) and the susceptibility to rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and ankylosing spondylitis (AS) among Chinese Han population in Lanzhou. Methods Primers for the two single nucleotide polymorphism (SNP) sites (rs805296 and rs805297) in ApoM gene were designed and their genotyping methods of polymerase chain reaction-high resolution melting (PCR-HRM) assay were established. Case-control studies were performed among the 599 cases of RA, 194 cases of SLE, 179 cases of AS and 273 matched healthy controls to analyze the correlations between the two SNPs and the susceptibility to rheumatic diseases. Results The genotype frequencies of rs805296 were AA 87.0%, AG 12.7%, GG 0.3% in RA cases, AA 84.5%, AG 15.0%, GG 0.5% in SLE cases, AA 91.6%, AG 7.3%, GG 1.1% in AS cases, AA 85.0%, AG 15.0%, GG 0% in healthy controls. The ones of rs805297 were GG 38.2%, GT 51.8%, TT 10.0% in RA cases, GG 44.3%, GT 45.4%, TT 10.3% in SLE cases, GG 37.4%, GT 47.5%, TT 15.1% in AS cases, GG 40.7%, GT 46.1%, TT 13.2% in healthy controls. Statistical analyses showed that only the genotype distribution of rs805296 was significantly different between the AS cases and the healthy controls. Under the dominant model, the G allele carriers of rs805296 (AG heterozygote and GG homozygote) were found to significantly decrease the risk for AS development. Conclusion The established PCR-HRM genotyping assays in the present study can successfully achieve the molecular diagnosis of the two SNPs sites (rs805296 and rs805297) from clinical samples, and the study found a significant association between the SNP of rs805296 and the susceptibility to AS among Chinese Han population in Lanzhou.

[High-resolution melting technology for detecting genetic polymorphisms of IL-1ß and IL-1Ra in patients with coronary artery disease].

OBJECTIVE: To develop a method of high-resolution melting curve analysis (HRM) for investigating the association of single nucleotide polymorphisms (SNPs) of IL-1ß and IL-1Ra genes with susceptibility to coronary heart disease (CHD) and serological indexes in CHD. METHODS: PCR-HRM assays were established for detecting four SNPs (IL-1B-511C>T, IL-1B-31C>T, IL-1B+3954C>T and IL-1RNT>C) in 260 patients with CHD and 274 healthy controls. A case-control study was performed for analyzing the relationships of the four SNPs with CHD risk and serological indexes. RESULTS: No significant statistical difference was observed between the four SNPs and the susceptibility to CHD, however, the result of gender stratification showed that there was the statistical differences in the male IL-1B-31C>T and the female IL-1B+3954C>T allele frequencies (P=0.039, 0.032, respectively). The result of haplotype analysis of the four SNPs showed that haplotype of IL-1B-511T/IL-1B-31C/IL-1B+3954C/IL-1RN T (TCCT) had a significant correlation with obviously increased CHD risk, but without statistical difference (OR=1.217, 95%CI: 0.950-1.559, P=0.123), while those of CTCC and TTCT had significant correlations with decreased CHD risk (OR=0.612, 95%CI: 0.376-0.994, P=0.046; OR=0.365, 95%CI: 0.154-0.862, P=0.017). No association was found between the four SNPs and the serological indexes in CHD subjects. CONCLUSION: The developed PCR-HRM detection assays based on HRM technology has been verified by direct sequencing and clinical specimen detection to be a rapid and homogeneous detection method for genotyping of the four SNPs. The SNPs of IL-1B-31C>T, IL-1B+3954C>T and the haplotypes of CTCC and TTCT are associated with the susceptibility to CHD in Chinese Han population of Lanzhou region.

Association analysis of single nucleotide polymorphisms of proinflammatory cytokine and their receptors genes with rheumatoid arthritis in northwest Chinese Han population.

OBJECTIVE: To analyze the relationship of genetic polymorphisms in IL1ß, IL6, TNF-α genes and their receptors genes with rheumatoid arthritis (RA) for northwest Han Chinese. This study also explores whether there are gene-gene interactions among these genetic polymorphisms. METHODS: A total of 452 patients with RA and 373 matched healthy controls were enrolled to carry out a case-control study for 16 SNPs of IL1B-511 C>T, IL1B-31 T>C, IL1B+3954 C>T, IL1RN T>C, IL6-597 G>A, IL6-572 G>C, IL6-174 G>C, IL6R-183 G>A, IL6R exon2 T>A, IL6R exon1 A>C, TNFA-863 C>A, TNFA-857 C>T, TNFA-308 G>A, TNFA-238 G>A, TNFR1-383 A>C and TNFR2 T676G T>G from seven genes. Genotyping for the SNPs was conducted on the RotorGene 6000 PCR platform using in-house high resolution melting (HRM) approaches. Detection correctness was validated through direct sequencing. Generalized multifactor dimensionality reduction (GMDR) analysis was applied to discover likely gene-gene interaction model among the SNPs. RESULTS: The results showed that the genotype distributions of TNFA-308, TNFA-857 and TNFA-863 are significantly different between case and control groups (P=0.016, P=0.048 and P=0.016, respectively). Carriers of TNFA-857 mutant allele conferred risk to RA (OR=1.525, 95% CI=1.157-2.009) while those of TNFA-308 and TNFA-863 mutant alleles conferred protection to RA (OR=0.459, 95% CI=0.286-0.739; OR=0.490, 95% CI=0.329-0.732). GMDR analysis for the SNPs indicated that gene-gene interaction existed among IL1B-31, IL1RN, IL6-572, IL6R-183, IL6R-exon1 and TNFA-857. Thirteen of all genotypes of the six SNPs combination were discovered to have significant distribution difference between RA group and the control. CONCLUSIONS: This study demonstrated that PCR-HRM assay is a highly efficient SNP genotyping method especially for the detection of large-scale samples. The SNPs of TNFA-308 and TNFA-863 are closely associated with RA susceptibility and that gene-gene interactions may occur among the six SNPs of IL1B-31, IL1RN, IL6-572, IL6R-183, IL6R-exon1 and TNFA-857 in RA patients from northwest Chinese Han population, especially these SNPs' combination genotypes CT/TT/CC/GG/AC/CC, CT/TT/GC/AA/AC/CT and CT/CT/CC/GA/AC/CC to show high risk of RA susceptibility in our study.