Abstract:Objective To study the role and effect of FEM1B in apoptosis of HIV reservoir ACH-2 cell lines. Methods A lentivirus used to overexpress FEM1B in ACH-2 cell lines. FITC-annexin-V/7-AAD staining was used to assess spontaneous cell apoptosis and cell apoptosis under TRAIL stimulation. Viral loads in culture supernatants were examined by qPCR. Results Spontaneous cell apoptosis remained unchanged in FEM1B-overexpressing cell lines. Cell apoptosis and viral loads in culture supernatants under TRAIL stimulation were significantly increased. Conclusions FEM1B increases TRAIL-mediated cell apoptosis and viral loads in culture supernatants of HIV reservoir cell lines.

Abstract:Objective To construct a recombinant luciferase reporter gene vector for wildtype and mutant 3’-untranslated regions (UTRs) of the LGALS3BP gene to study regulation of the LGALS3BP gene and miRNAs. Methods TargetScan was used to predict the binding site of miRNAs in the 3’UTR of the LGALS3BP gene. The wildtype LGALS3BP 3’UTR was amplified by PCR using genomic DNA from SMMC-7721 hepatocellular carcinoma cells as a template. PCR products were digested by two restriction enzymes and inserted into the luciferase reporter vector pGL3-control. Recombinant plasmids were verified by colony PCR and sequencing. Primers for LGALS3BP 3’ UTR with binding site mutations were designed. Single and double site mutant luciferase reporter gene vectors were prepared by overlapping PCR and site-directed mutagenesis PCR using the wildtype LGALS3BP 3’UTR recombinant plasmid as a template. Results Colony PCR showed a target gene fragment of 871 bp, which was consistent with the expected size. The inserted sequence was consistent with the LGALS3BP 3’ UTR in sequencing. TargetScan indicated two miRNA-binding sites in the LGALS3BP 3’UTR. The single and double site mutant sequences were successfully inserted into the LGALS3BP 3’UTR. Conclusions Wildtype LGALS3BP 3’UTR (pGL3-LGAL-W-3’UTR), two single site mutants (pGL3-LGAL-M1-3’UTR and pGL3-LGAL-M2-3’UTR), and a double site mutant (pGL3-LGAL-M-3’UTR) luciferase reporter gene vectors were successfully constructed, which may facilitate studies of LGALS3BP gene regulation through miRNAs.

Abstract:Objective To establish a rat model of chronic kidney disease with hyperhomocysteinemia by high methionine feeding and 5/6 nephrectomy. Then, to evaluate the model by comparing it with the simple high methionine feeding model and simple nephrectomy model. Methods Five-week-old rats were randomly divided into four groups: sham, high methionine (feeding on a high methionine diet), nephrectomy (5/6 nephrectomy), and compound model (high methionine diet and 5/6 nephrectomy) groups. Arterial blood pressure, echocardiography, and serum biochemical indices were measured. Changes in the tension of isolated thoracic aortic vascular rings were observed and expression of pp85, p-Akt, and p-eNOS (Ser1177) proteins was detected. HE staining was used to observe pathological changes in renal, myocardial, and aortic tissue. Results　Compared with the sham group, arterial blood pressure of the three model groups increased continuously (P<0.01). Arterial blood pressure of compound model group was significantly higher than high methionine group and nephreetomy group(P<0.05). Compared with the sham group, echocardiography indexes LVEDd, LVEDV, LVESV, and SV of the three model groups were significantly different (P<0.05). Compared with the sham group, serum homocysteine and methionine in high methionine and compound model groups were increased (P<0. 01), while, creatinine, urea nitrogen, total cholesterol, and high-density lipoprotein in serum were increased significantly (P<0. 05) and triglyceride was significantly decreased (P<0. 05) in nephrectomy and compound model groups. Compared with the sham group, vascular ring relaxation curves of model groups induced by vasodilators shifted to the right, and the Emax and EC50 of the relaxation effect induced by acetylcholine and sodium nitroprusside in the compound model group were significantly different from those in the simple model group (P<0. 05). Compared with the sham group, expression of p-Akt and p-eNOS in aortic tissue of the compound model group was statistically significant (P<0. 05). Pathological changes were observed in renal, myocardial, and aortic tissues of the model animals, among which the compound model group had more severe pathological changes. Conclusions A rat model of chronic kidney disease with hyperhomocysteinemia can be established by combining a high methionine diet and 5/6 nephrectomy. Compared with the simple models, the functional and morphological changes are significantly different in the compound model, which can be applied to fundamental research on chronic kidney disease with hyperhomocysteinemia.

Abstract:Objective To analyze the serum metabolomics of non-alcoholic fatty liver disease (NAFLD), type 2 diabetes mellitus (T2D), and atherosclerosis (AS). Methods NAFLD, T2D, and AS mouse models were established, and serum metabolomics were analyzed by ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS). Data were analyzed using the R language MetaboAnalystR package. Differential metabolites were screened by multivariate statistics. Differential metabolic pathways were obtained by untargeted KEGG enrichment analysis. The result were analyzed for correlations between differential metabolites and blood lipids. Results NAFLD, T2D and AS shared 48 differential metabolites that were particularly enriched in linoleic acid metabolism, pentose phosphate metabolism, arachidonic acid metabolism, and histidine metabolism pathways (P<0. 05). The differential metabolites were significantly correlated to blood lipids. Compared with the control group, the main specific metabolic pathways of NAFLD, T2D and AS were Darginine and D-ornithine metabolism (P=0. 09), galactose metabolism, primary bile acid biosynthesis, starch, and sucrose metabolism (P<0. 05), and sphingolipid and pyrimidine metabolism (P<0. 05), respectively. The three diseases were compared pairwise, and there were differences in D-arginine and D-ornithine metabolism (P=0. 12), starch and sucrose metabolism (P<0. 01), D-glutamine and D-glutamate metabolism (P<0. 05), synthesis and degradation of ketone bodies (P=0. 06), alanine, aspartate and glutamate metabolism (P<0. 05), sphingolipid metabolism (P<0. 05), valine, leucine and isoleucine degradation (P<0. 05), and glycine, serine, and threonine metabolism (P=0. 12). Conclusions This study revealed the common and specific metabolic characteristics of NAFLD, T2D and AS, and provides for a research direction for comprehensive prevention and individualized treatment of these diseases.

Abstract:Objective Optimization of the surgical details of acute myocardial infarction in neonatal 1-day-old mammary mice, and establishment standardized assessment of a stable model. Methods Postnatal day 1 (P1) CD1 mice were subjected to three sets of surgical manipulations. Coronary artery left anterior descending (LAD) ligation (1 mm below the left auricular border, 1 mm in width, and <0. 5 mm ligation depth) was performed in the standard myocardial infarction (SMI) group. The LAD ligation position and width remained unchanged with a >0. 5 mm ligation depth in the deep ligation group (Deep MI, DMI). Only the chest was opened without LAD ligation in the sham surgery group (Sham). The success of LAD ligation was verified by TTC and Evans blue-TTC staining. The extent of myocardial tissue damage, fibrosis, and regeneration of LAD ligation was assessed by HE and Masson staining at 3, 7, 14, 21 and 28 days after surgery. Structural and functional changes of mouse hearts were assessed by echocardiography at 28 days after surgery. Results It describes the process of establishing a P1 neonatal mouse myocardial infarction model in detail and provides stable regeneration and a high survival rate of MI through realization of LAD exposure, ligation, postoperative care, and other processes. Successful ligation of the SMI model was determined by TTC and Evans blue-TTC staining at 1 day postoperatively. Echocardiography at 28 days postoperatively revealed no statistical difference in LVEF, LVFS, LVIDd, or LVIDs compared with the Sham group, indicating that the heart structure and function were restored to normal. Masson staining at 3, 7, 14, 21 and 28 days postoperatively revealed that tissue fibrosis had occurred in 15. 67%, 3. 34%, 2. 99%, 2. 73% and 1. 11% of the heart area, respectively, in the SMI group, indicating almost complete recovery of the myocardial infarct area at 28 days postoperatively. The survival rate of the DMI group was reduced by 35. 71% compared with the SMI group (85. 71% for SMI and 50% for DMI). Ultrasound at 28 days postoperatively showed reduction in LVEF (17. 25±6. 03)%, LVFS (11. 37±4. 06)%, increases in LVIDd (0. 46±0. 15)%, and LVIDs (0. 69±0. 20)% (P<0. 05). The fibrotic area was six times larger in the DMI group than in the SMI group, indicating that complete regenerative repair at 28 days postoperatively could not be achieved with a ligation depth of > 0. 5 mm. Conclusions This study describes in detail the process of establishing a standard acute myocardial infarction model in 1-day-old mice, and evaluated the infarct size and cardiac structure, function, and fibrosis at various time points after surgery by TTC, Evans blue-TTC staining, cardiac ultrasound, and Masson staining. It was clear that complete cardiac repair at 28 days after surgery cannot be achieved with a ligation depth of >0. 5 mm. These data provide a reference to establish a stable and reliable model of acute myocardial infarction in 1-day-old mice for surgical evaluation.

Abstract:Objective To investigate the effect of chiropractic therapy on the inflammatory response and apoptosis of intervertebral disc cells in rats with cervical spondylosis and its possible mechanism. Methods Overall, 8 SPF SD rats were randomly selected as the sham operation group (half male and female). The remaining rats were used to establish an intervertebral disc model by the dynamic and static imbalance method . Model rats were randomly divided into model, chiropractic, and control groups with eight rats in each group. The chiropractic group was subjected to chiropractic therapy every day, the control group was administered 0. 75 mg/ kg meloxicam tablets every day, and sham operation and model groups were untreated for 28 days. HE staining was used to detect histopathological changes of the intervertebral disc. TUNEL staining was used to detect apoptosis of intervertebral disc cells. Immunohistochemistry was used to detect tumor necrosis factor (TNF-α) and interleukin-1β (IL-1β) protein expression in intervertebral disc tissue. Serum TNF-α and IL-1β levels were measured by ELISA. RT-qPCR was used to measure relative mRNA expression of Caspase-3, B-cell lymphoma/ leukemia-2 gene (Bcl2) and Bcl2-related X protein (Bax). Protein expression of Cleaved Caspase-3, Bax, Bcl2 and IκB kinase β (IKKβ), p-IKKβ, NF-κB inhibitor protein α (IκBα), p-IκBα, nuclear factor-κB p65 (NF-κB p65) and p-p65 was detected by Western blot. Results Compared with the sham operation group, the model, chiropractic, and control groups showed that the tissue structure of intervertebral disc was in various degrees of degeneration, the nucleus pulposus tissue shrinked, and the boundary with the external fibrous ring was unclear, meanwhile, intervertebral disc histopathological score, nucleus pulposus (NP) cell apoptosis rate, serum TNF-α and IL-1β, TNF-α and IL-1β protein expression, Caspase-3 and Bax mRNA expression, and p-IKKβ, p-p65 and p-IκBα protein expression in the intervertebral disc tissue all increased, while the expression of Bcl2 mRNA and protein decreaed (P<0. 05). Compared with the model group, the chiropractic, and control groups showed that degeneration of the intervertebral disc tissue structure was reduced gradually, and the intervertebral disc histopathological score, NP cell apoptosis rate serum TNF-α and IL-1β, TNF-α and IL-1β protein expression, caspase-3 and Bax mRNA expression, and p-IKKβ, p-p65 and p-IκBα protein expression in the intervertebral disc tissue were decreased, while Bcl2 mRNA and protein expression were increased in intervertebral disc tissue (P<0. 05). Changes in the above indexes and factors in the chiropractic group were lower than those in the control group (P<0. 05). Conclusions Chiropractic therapy reduces the inflammatory reaction and apoptosis of intervertebral disc cells in rats with cervical spondylosis and improves the progression of intervertebral disc degeneration. The mechanism may be related to inhibition of the IKKβ/ NF-κB pathway.

Abstract:Objective To investigate the ameliorative effect and possible mechanism of Danshensu in retinal vein occlusion (RVO) rats. Methods Eight SPF grade male SD rats were randomly selected as the control group. The RVO model was induced by Bengal red combined with the laser photodynamic method in the left eye of the remaining rats. The model rats were randomly divided into model, Danshensu low (15 mg/ kg), medium (30 mg/ kg), and high dose group(60 mg/ kg), with eight rats in each group. Rats in control and model groups were injected with the same amount of normal saline through the caudal vein every day for 21 days. Fundus fluorescein angiography was used to observe the retinal vein structure. The venous blood flow velocity was measured by ear vein microcirculation. A full field electroretinogram(ffERG) was used to evaluate retinal functions. The retinal layer thickness was measured by optical coherence tomography. Histological changes of the retina were observed by HE staining. The relative expression of VEGF mRNA was measured by RT-qPCR. Expression of hypoxia inducible factor 1α (HIF1α), extracellular signal regulated kinase 1/2 (ERK1/2), p-ERK1/2, focal adhesion kinase (FAK), p-FAK, VEGF, and pigment epithelium-derived factor (PEDF) in retinal tissue was detected by Western blot. Results Before administration, compared with the control group, the retinal vein blood flow of model rats was interrupted, distal blood vessels were tortuous and dilated, the retinal thickness was increased, the amplitudes of ffERG a and b waves were decreased, and the blood flow velocity of the ear vein was decreased (P<0. 05). After administration, compared with the control group, retinal vein occlusion was recanalized, the retinal thickness, amplitudes of ffERG a and b waves, and blood flow velocity of the auricular vein were decreased (P<0. 05), and the retina was damaged by various degrees. The retinal structure of the model group was incomplete, the number of ganglion cell layer cells was decreased, the photoreceptor cell layer of the outer nuclear layer was almost lost, and relative expression of VEGF mRNA and HIF1α, p-ERK1/2, and p-FAK proteins was increased in retinal tissue, while the relative expression of PEDF protein was decreased (P<0. 05). Compared with the model group, the changes in the above indexes and factors in the three Danshensu treatment groups were reversed (P<0. 05) and pathological damage of the retina was gradually reduced. The effect of Danshensu was dependent on dose (P<0. 05). Conclusions Danshensu reduces retinal tissue injury, promotes recovery of blocked venous microcirculation, and improves retinal functions in RVO rats. Its mechanism may be related to inhibiting activation of VEGF-related angiogenesis pathways.

Abstract:Objective To investigate whether Baicalein (BAI) protects nucleus pulposus cells (NPCs) exposed to tert-Butyl hydroperoxide (TBHP). Methods NPCs were extracted from 8-week-old mice and cultured to passages 2 and 3 for experiments in control, model, and baicalein groups. SA-β-galactosidase staining was used to detect senescence of NPCs. Real-time quantitative PCR was used to measure aggrecan, collagen Ⅱ, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-9 (MMP-9), tumor necrosis factor α (TNF-α), interleukin (IL)-1β, p53 and p16 mRNA expression. TNF-α was detected by immunofluorescence. Protein expression of aggrecan, p53, p16, IL-1β and TNF-α was detected by Western blot. Results The concentration of TBHP was 100 μmol/ L, and the treatment time was 4 h. The optimal concentration of baicalein was 1 μg/ mL with a treatment time of 24 h. Compared with the control group, the positive rate of SA-β-galactose staining in the TBHP group was significantly increased (P<0. 05), p53, p16, IL-1β and TNF-α mRNA and protein expression was significantly increased ( P<0. 05), and MMP-3 mRNA expression was significantly increased (P<0. 05). MMP-9 mRNA expression was increased, but the difference was not statistically significant (P>0. 05), aggrecan mRNA and protein expression was significantly decreased (P<0. 05), collagen Ⅱ mRNA expression was decreased, but the difference was not statistically significant (P>0. 05), and IL-6 protein expression was increased, but the difference was not statistically significant (P>0. 05). Compared with the model group, the positive rate of SA-β-galactose staining in the baicalein group was significantly lower (P<0. 05). p53, p16, IL-1β, and TNF-α mRNA and protein expression was significantly lower (P<0. 05), and MMP-3 and MMP-9 mRNA expression was significantly lower (P<0. 05). Aggrecan mRNA and protein expression was significantly increased (P<0. 05), collagen Ⅱ mRNA expression was significantly increased (P<0. 05), and IL-6 protein expression was significantly decreased (P<0. 05). Conclusions Baicalein delays senescence of NPCs induced by TBHP.

Abstract:Objective To establish and evaluate a mouse model of inhaled anthrax infection by Bacillus anthracis(Pasteur II strain) spores by aerosolized intratracheal inoculation. Methods B10. D2-Hc0H2d H2-T18c / oSnJ mice lacking complement component C5 were infected with Bacillus anthracis Pasteur II strain spores (attenuated strain) by aerosolized intratracheal inoculation. Disease progression, tissue bacterial load, histopathological changes, cytokine response, and acute reactive protein levels were examined after infection. Results The median lethal dose was 5×103 CFU. Multiple organ metastasis and bacteremia occurred within 12 hours after pulmonary infection of anthrax spores. Histopathological changes and inflammatory responses were similar to those in other species including rabbits, nonhuman primates, and humans. C-reactive protein and amyloid protein P levels in serum and alveolar lavage fluid indicated that acute inflammatory responses were triggered after infection. Conclusions This model can be used as an alternative small animal model to further elucidate the pathogenesis of Bacillus anthracis and differences in host susceptibility to B. anthracis, providing new approaches for anthrax vaccines and therapeutic drug development.

Abstract:Programmed cell death protein 1 (PD-1) is an important immune checkpoint molecule that regulates the immune system and promotes autoimmune tolerance by regulating the immune response to human cells and inhibiting the inflammatory activity of T cells. Recent studies have found that PD-1 plays an important role in regulating maternal and fetal immune tolerance and maintaining pregnancy. This article reviews the source, distribution, and biological characteristics of PD-1 and its role in normal pregnancy as well as the research progress of abnormal PD-1 expression in the pathogenesis of pregnancy-related diseases such as pre-eclampsia and recurrent abortion to provide a theoretical basis for early prevention, early diagnosis, and timely treatment.

Abstract:Perimenopausal depression (PMD) is a kind of mental disease that occurs during the perimenopausal period accompanied with typical clinical manifestations of emotional depression, anxiety, and thinking retardation. The pathogenesis of PMD mainly includes hormone levels, neurotransmitters, inflammatory reactions, and brain-derived neurotrophic factor. It has been suggested that Traditional Chinese medicine affects PMD. In this review, the pathogenesis of PMD is discussed in order to provide a reference for mechanistic research and treatment of PMD.

Abstract:Depression is a common mental disorder with a high recurrence and suicide rate and is a serious threat to human physical and mental health. Animal models of depression mimic the disease phenotype of human depression, helping humans to study the pathogenesis of the disease and develop new antidepressant drugs. Although several rodent models of depression have been established, none of the existing single models simulate the whole disease well. The development of compound models allows researchers to more comprehensively analyze depression with higher reliability, but there are also problems such as complicated practical operations and poor consistency. Therefore, to select animal models that better meet experimental requirements, we reviewed articles related to rodent models of depression published up to 2021, compared the frequency of use of each model in the previous 5 years, and comprehensively summarized their modeling method , reliability evaluation, advantages and disadvantages, and current applications from the etiological and pathophysiological mechanisms of depression. We also systematically reviewed the current commonly used rodent models of depression. Additionally, the current rodent models of depression, including stress, pharmacological, genetic, surgical injury, composite, and other models, were systematically reviewed. Challenges in the establishment and use of future rodent models of depression are also presented to provide researchers with more feasible references, preferred options, and innovative directions to model depression.

Abstract:With the high incidence and frequency of depression, research on regulation of depression by the inflammatory system has become a hot topic. As an important means to improve depression, current research has confirmed that proinflammatory factors, such as the interleukin-1 family, and TNF-α, and TGF-β, through influencing HPA axis activity, participate in the mechanism of regulating depression, and exercise improves the occurrence and development of depression. However, research on the mechanism of exercise mediated by the inflammatory system in improving depression needs to be explored. Therefore, this review explored the role of the inflammatory system in depression and its important regulatory role in exercise to improve depression from the aspects of proinflammatory and anti-inflammatory cytokines. This will contribute to a more comprehensive understanding of the role of the inflammatory system in the occurrence of depression, and provides new ideas and perspectives to study the mechanism of exercise to improve depression.

Abstract:FEM1B is a highly conserved gene, and the protein encoded by FEM1B is a member of the anchorrepeat protein family. Its spatial conformation contains 7 modules in series. FEM1B is involved in the regulation of a variety of biological functions, including apoptosis, protein ubiquitination modification, cell reductive stress and DNA replication stress. FEM1B has been shown to promote apoptosis of tumor cells in colorectal cancer, human T-cell acute lymphoblastic leukemia, and diffuse large B-cell lymphoma, while downregulation of FEM1B inhibits proliferation, invasion, and migration of tumor cells in non-small cell lung cancer. In addition, the structure of FEM1B is related to the HIV regulatory protein Vif. These characteristics of FEM1B may make it a new target for cancer treatment and the regulation of HIV infection.

Abstract:Environmental air pollution causes serious damage to the human respiratory system and increases the risk of respiratory diseases. Smoke stimulation is a modeling method commonly used in clinical animal experiments to prepare animal models of respiratory diseases, including cigarette smoke stimulation and thick smoke stimulation, by imitating the pathological principle of air environmental pollution. Through drug treatment of animal models, researchers can develop drugs to treat respiratory diseases and alleviate symptoms. Research on the mechanism of traditional Chinese medicine and external treatment with traditional Chinese medicine for respiratory diseases is constantly progressing. Traditional Chinese medicine therapy has unique advantages in the treatment of respiratory diseases. Therefore, establishing an animal model with good reproducibility and high consistency with clinical respiratory diseases is particularly important to prevent, treat, and research respiratory diseases. This article mainly discusses harm caused by environmental air pollution to the human respiratory system, expounds the pathological mechanism of the smog stimulation method and the preparation elements and method of animal models, and analyzes the application of animal models established by the smog stimulation method in traditional Chinese medicine and external treatment with traditional Chinese medicine, laying the foundation to establish more reasonable animal models of respiratory diseases and the emergence of new therapies and drugs in traditional Chinese medicine.

Abstract:Threatened abortion complicated with intrauterine hemorrhage is a common disease during pregnancy. Its pathogenesis is related to an imbalance of the maternal-fetal interface microenvironment and obstruction of uterine spiral artery remodeling. Trophoblast cells compose the outermost layer of the maternal-fetal interface microenvironment. Their proliferation, migration, and invasion are closely related to normal pregnancy. They play an irreplaceable role in immune tolerance and regulation, uterine spiral artery remodeling, and maintenance of the maternal-fetal interface microenvironment. In recent years, trophoblast cells have become the main research focus in autoimmune diseases and other fields. Studies have shown that trophoblast cells play an important role in the diagnosis and treatment of threatened abortion complicated with intrauterine hemorrhage. This article reviews the research progress of trophoblast cells in the treatment of threatened abortion complicated with intrauterine hemorrhage.

Abstract:Depression is a disease that seriously endangers human physical and mental health. The establishment of a suitable animal model of depression is of great significance to study its pathogenesis and develop drugs. Among the animal models of depression, animal models of drug-induced depression are widely used because of the advantages of simplicity and timely establishment. This article systematically reviewed drug-induced depression animal models in the China National Knowledge Network and PubMed until July 1, 2021, and reviewed six commonly used drug-induced depression animal models. The frequency of use of induced depression animal models, selection of model animals, drug injection dosage, injection method , indicators for model evaluation, advantages, and disadvantages are expected to provide references to prepare depression models and develop drugs.

Abstract:To summarize the animal and cell models of attention deficit hyperactivity disorder (ADHD) and describe its pathological mechanism to provide a theoretical foundation for experimental pharmacological research. Research on ADHD was gathered from CNKI, Wanfang, PubMed, and other databases. The principles, characteristics, and research progress of ADHD animal models were comprehensively outlined, and their merits and shortcomings were highlighted. The research status of ADHD cell models was evaluated and the pathological mechanism of ADHD was explained. There are two types of animal models for ADHD: genetic models (hybrid mouse and transgenic animal models) and non-genetic models (chemically and environmentally induced models). The focus on face validity, construct validity, and prediction validity differs among models in addition to their application span. Neuronal and non-neuronal cell lines are the most often used cell lines for ADHD research, but they are understudied. The etiology of ADHD mostly involves neurotransmitter dysregulation, neuroinflammation, and decreased neuronal energy. SHR is the optimum animal model to simulate ADHD symptoms and has a strong predictive validity for neurostimulator medications. SH-SY5Y cells display the dopaminergic phenotype of ADHD and are a useful tool to determine cell viability and drug safety. Neurotransmitter dysregulation, particularly dopamine insufficiency, appears to be the primary pathophysiology of ADHD in accordance with in vitro and in vivo investigations.