Abstract:Objective To explore the effects of miR-124-3P on the proliferation and invasion of placental trophoblasts during preeclampsia by targeting MAPK 14. Methods Preeclampsia rat models were established, followed by extraction of placental trophoblast tissue and primary cells from normal rats and model rats. Primary cells were manipulated by transfections. The MTT assay was used to determine cell viability; flow cytometry was used to detect apoptosis and cell cycle arrest; Transwell was used to determine the cell migration and invasion ability, and qRT-PCR and western blotting were used to determine the expression of miR-124-3P and MAPK 14 as well as the expression level of migration-related genes. Verify the binding relationship between miR-124-3P and MAPK 14. Results Compared with the control cells, miR- 124-3p overexpression reduced cell proliferation and invasion. Cells arrested at the G1 / G0 phase, and the apoptosis rate increased. Suppressing miR-124-3P expression produced opposite result . The dual luciferase reporter experiment showed that MAPK 14 was the target of miR-124-3P. Suppressing the expression of MAPK 14 produced result that were consistent with the miR-124-3Poverexpression effects, i.e., the cell viability and invasion ability decreased, and the apoptosis rate increased. In contrast, MAPK 14 overexpression in the experimental group was consistent with the effects of inhibition of miR-124-3P expression. Conclusions miR-124-3P negatively regulates MAPK 14 and affects the proliferation and invasion of placental trophoblasts in preeclampsia rats.

Abstract:Objective To investigate the effect of the G protein-coupled estrogen receptor (GPER) on oxidative stress and its possible mechanisms for reducing renal ischemia reperfusion injury. Methods Female ovariectomized rats were divided into the ovariectomy (OVX), renal ischemia reperfusion (OVX+I/ R), estrogen intervention (OVX+I/ R+E₂ ), GPER-specific agonist (G1) intervention (OVX+I/ R+G1), and estrogen+GPER specific blocker G15 intervention (OVX+I/ R+G15+ E₂ ) groups ( n= 8 rats per group). Serum creatinine ( Cr) and urea nitrogen ( BUN) levels were measured, and histopathological examination using hematoxylin and eosin ( HE) staining was used to observe the pathological morphology of the renal tissues. Superoxide dismutase and malondialdehyde activities were detected, and Western blot was used to observe p-PI3K and p-Akt expressions in the renal tissues of each group. Results Compared with the OVX group, serum Cr and BUN levels were increased, pathological damage to the renal tissue was significant, oxidative stress was aggravated, and p-PI3K and p-Akt protein expression levels were significantly decreased in the OVX+I/ R group (allP< 0. 01). Compared with the OVX + I/ R group, the serum Cr and BUN levels were decreased, renal tissue pathological damage was reduced, oxidative stress was reduced, p-PI3K and p-Akt protein expressions were increased (all P< 0. 01), and the GPER-specific inhibitor, G15, partially eliminated the protective effect of E₂ on renal ischemia reperfusion injury in the E₂ and G1 intervention groups. Conclusions During renal ischemia reperfusion injury, E₂ and G1 may reduce oxidative stress to alleviate the injury. GPER may mediate the protective effect of E₂ , and its mechanism may involve activating the PI3K/ Akt signaling pathway.

Abstract:Objective Whether porcine circovirus type 2 (PCV2) can infect human cells is controversial. To study the antiviral effects of human cells on PCV2 and other viruses, we analyzed differentially expressed genes (DEGs) and their related signaling pathways and biological functions in PCV2-infected human cells by transcriptome analysis (RNA sequencing, RNA-seq). Methods HeLa cells were infected with PCV2, and then analyzed by RNA-seq to screen DEGs related to antiviral reactions. The DEGs were further verified by Real-time quantitative PCR. Results A total of 15402 UniGenes and 387 DEGs, including 267 upregulated genes and 120 downregulated genes, were annotated in human HeLa cells infected with PCV2. The most abundant Gene Ontology (GO) terms were response to stress ( 97 / 291 DEGs) and immune system process (80 / 291 DEGs). The most significant pathways were the nucleotide-binding and oligomerization domain ( NOD)-like receptor signaling pathway ( 21 / 170 DEGs ) and herpes simplex infection ( 22 / 170 DEGs ). Conclusions The gene expression profile of HeLa cells significantly changed after PCV2 infection. The result obtained in this study will help understand the antiviral mechanism of human cells and may provide a basis for understanding the immune response and antiviral efficacy of human cells infected with PCV2.

Abstract:Objective To investigate the effects of isoliquiritigenin ( ISO) on cisplatin-induced acute kidney injury (AKI) in mice and explore the potential mechanism. Methods Thirty C57BL/ 6 male mice were randomly divided into a blank control group, model group, ISO-L and ISO-H groups and a positive control group. A single intraperitoneal injection of cisplatin (20 mg / kg) was applied to establish the AKI model. During the intervention, both blank control group and model group were treated with an equivalent volume of saline, while ISO-L group and ISO-H group were administered by gavage with 7. 5 mg / kg and 30 mg / kg ISO, respectively, while the positive control group was administered by gavage with 20 mg / kg irbesartan. After 3 days, serum was collected to determine the levels of serum creatinine ( SCr) and blood urea nitrogen (BUN). HE and PAS staining were used to detect pathological kidney changes, while immunohistochemical and western blot analyses were used to detect inflammatory cytokines’ ( IL-6 and IL-1β) expression and the activity of Smad3 and NF-κB. Real-time PCR was used to detect changes in inflammatory cytokines and long noncoding Arid2-IR. Results Compared with the model group, ISO intervention significantly reduced the SCr and BUN levels in AKI mice (P< 0. 05), in a concentration-dependent manner. Histopathological staining indicated that ISO intervention significantly reduced the infiltration of inflammatory cells and vacuolar degeneration of renal tubular epithelial cells. Furthermore, it significantly improved renal injury. Additionally, ISO significantly downregulated (P< 0. 05) inflammatory cytokines expression, the activity of Smad3 and NF-κB, and long noncoding Arid2-IR expression, suggesting that ISO inhibited the activation of the Smad3 / Arid2-IR/ NF-κB axis. Conclusions ISO effectively reduced kidney inflammation in AKI mice, possibly through regulation of the Smad3 / Arid2-IR/ NF-κB axis.

Abstract:Objective To explore the inhibitory effect of Zhenqi Fuzheng granules on angiogenesis in hepatocellular carcinoma (HCC) model rats and the possible mechanisms. Methods Wistar rats were divided into the control, model [discontinuous low-dose (Diethyl ammonium nitrite, DEN) induction method , Zhenqi Fuzheng granule (2. 62 g / kg), si-miR-200c (20 pm), and Zhenqi Fuzheng granule + si-miR-200c groups ( n= 12 rats per group) and were treated for 8 weeks. Hematoxylin and eosin staining was used to observe the pathomorphological changes in the rat liver tissues. Serum vascular endothelial growth factor ( VEGF ), alanine aminotransferase ( ALT ), and aspartate aminotransferase (AST) levels were detected via enzyme-linked immunosorbent assay ( ELISA). Real-time fluorescence quantitative PCR was used to detect miR-200c expression in the liver tissues; zinc finger protein ( ZEb-2) and VEGF protein were detected via western blot, and the positive expressions of ZEb-2, VEGF and microvessel density (MVD) were detected via immunohistochemistry. Results Compared with the control group, VEGF, ALT and AST levels were increased in the model group, and the model group livers contained more polymorphic and necrotic cells and showed decreased miR-200c expression, increased ZEb-2 and VEGF expressions, and increased ZEb-2, VEGF and MVD positivity rates (allP< 0. 05). Compared with the model group, the Zhenqi Fuzheng granule group had lower VEGF, ALT and AST levels, reduced cell deformation, higher cancer cell differentiation, increased miR-200c expression, decreased ZEb-2 and VEGF expressions, and decreased ZEb-2, VEGF and MVD positivity rates. The si-miR-200c group presented increased VEGF, ALT and AST levels, increased cell carcinogenesis, decreased miR-200c expression in the liver, increased ZEb-2 and VEGF expressions, and increased ZEb-2, VEGF and MVD positivity rates ( allP< 0. 05). VEGF, ALT and AST levels in the Zhenqi Fuzheng granule + si-miR-200c group were lower than those in the si-miR-200c group, while the ZEb- 2 and VEGF expressions and the ZEb-2, VEGF and MVD positivity rates were lower, and the miR-200c expression was higher than that of the si-miR-200c group (allP< 0. 05). Conclusions Zhenqi Fuzheng granules inhibited angiogenesis and alleviated HCC development in rats. Its mechanism may be related to upregulating miR-200c expression and downregulating ZEb-2 protein expression.

Abstract:Objective To use CRISPR/ Cas9 gene editing technology to efficiently construct Bpi gene-knockout mouse models and continue to breed, identify and establish stable genetic Bpi gene-knockout mouse strains. Methods Knockout targets were designed at the two ends of exons 2-3 of the Bpi gene in C57BL/ 6 mice, and high activity guide RNA ( gRNA) targets were selected. The small guide RNA(sgRNA)was transcribed in vitro and microinjected into fertilized eggs of C57BL/ 6 mice together with Cas9 mRNA. F0 mice were obtained after embryo transfer into the surrogate mice. Positive mice with gene mutation was confirmed via genotype identification and DNA sequencing. Results Microinjection with Cas9 mRNA yielded highly active sgRNA and 64 fertilized eggs in good condition, which were successfully transplanted into 2 surrogate mice, yielding 22 F0 mice. After PCR identification and DNA sequencing, a mouse that was positive for a 708 bp single- strand deletion was selected to be propagated. The deletion was detected in the F1 and F2 generations, and homozygous mice were obtained in the F2 generation. Conclusions Bpi-knockout mouse models were successfully constructed, laying a foundation for further study of the biological functions of the Bpi gene and its expression products.

Abstract:Objective To study the absorption mechanism and improve the bioavailability of ambroxol hydrochloride (AMB) across the Caco-2 cell monolayer model. Methods The integrity and functionality of the Caco-2 cell monolayer model were analyzed by the transepithelial electrical resistance (TEER), the apparent permeability coefficient (Papp ) of acyclovir and the efflux ratio (ER) of rhodamine 123. The effects of drug concentration, drug-drug interaction and pH on the P_app(AP→BL) and ER of AMB were investigated using the Caco-2 cell monolayer model. Results The TEER values of all the permeation experiments were above 200 Ω·cm2 . The P_app(AP→BL) of acyclovir was less than 1×10^-6 cm/ s. Furthermore, the ER of rhodamine 123 was lower than 2, indicating that the Caco-2 monolayer model was successfully established in this experiment. Compared with the high permeability control group, the P_app(AP→BL) was significantly decreased in the low-dose AMB group, but the ER was not affected. There was also no difference in the ER between AMB used alone and in combination with METO; however, there was a significant increase in the P_app(AP→BL) in the monotreatment group. Furthermore, the P_app(AP→BL) of AMB transport under pH 6. 5 showed a significant difference compared with that under pH 7. 4 and 8. 0 (P< 0. 01). Conclusions The transport of AMB across the Caco-2 cell monolayer model was facilitated by diffusion, and combination therapy was not conducive to AMB absorption. However, an alkaline environment accelerated AMB absorption.

Abstract:Objective To investigate the effect of betulin on Aβ_25-35 -induced oxidative stress and apoptosis of PC12 cells. Methods PC12 cells were treated with 40 μmol / L of Aβ_25-35 . The cells were randomly divided into control group, Aβ_25-35 group, NAC group, and betulin 5, 10 and 20 μmol / L groups. Apoptosis was detected by V-FITC/ PI double staining flow cytometry; mitochondrial membrane potential was detected by JC-1 kit; LDH, MDA and SOD levels were detected by LDH, MDA and SOD kits, respectively; the enzyme activity of caspase-3, caspase-8, caspase-9 and Cyt c was detected by colorimetry; the activity and expression of Bcl 2, Bax, caspase-3, caspase-8, caspase-9 and Cyt C were detected by western blotting. Results Compared with the Aβ_25-35 group, betulin and NAC improved the cell survival rate and SOD activity ( P < 0. 01), reduced the LDH release rate and apoptosis rate ( P < 0. 01), inhibited caspase - 3, caspase-8, caspase-9 and Cyt c enzyme activity (P < 0. 01), and downregulated Bax, caspase-3, caspase-8, caspase- 9 and Cyt c enzyme activity (P < 0. 01) and expression level (P < 0. 01). In contrast, these treatments upregulated the activity and expression of Bcl2 ( P< 0. 01 ). Conclusions Betulin inhibited the Aβ_25-35 -induced apoptosis. The mechanism may be related to the reduction in ROS production, thus reducing the level of oxidative stress, and thereby inhibiting apoptosis.

Abstract:Objective To investigate the effect of glycyrrhizin compound (CG) on mouse models of experimental autoimmune encephalomyelitis (EAE) via the JAK2 / STAT3 / SOCS3 signaling pathway. Methods Fifty female C57BL/ 6 mice were randomly divided into the control group, model group, and high-dose, medium-dose and low-dose CG intervention groups (n = 10 mice per group). The model and CG intervention groups established the EAE model. The intervention groups were injected with CG (15 mg / (kg·d), 30 mg / (kg·d), or 60 mg / (kg·d)) for 14 consecutive days. The control and model groups were simultaneously intraperitoneally injected with equal volumes of saline. The neurological deficit scores and pathological changes in each group were recorded. Western blot was used to detect protein expression in the brain tissue, and real-time fluorescence quantitative RT-PCR was used to detect mRNA expression in the brain tissue. Results Compared with the model group, the highest neurological deficit and cumulative neurological deficit scores were reduced, spinal cord inflammation and demyelination were reduced, JAK2 and STAT3 protein and mRNA expressions were decreased, SOCS3 protein and mRNA expressions were increased, RORγt mRNA expression was decreased, and FOXP3 mRNA expression was increased in all CG groups ( all P < 0. 05). Conclusions CG exerted preventive and therapeutic effects on EAE model mice. The mechanism may be related to upregulation of SOCS3, inhibition of the JAK/ STAT signaling pathway, and rectification of the Th17 / Treg immune imbalance.

Abstract:Objective To explore the effects of Tet1 protein on cell proliferation, the cell cycle and apoptosis, an inducible Tet1CD overexpression cell line was established. Methods A recombinant pTRE-Tight_Tet1CD_IRES_RFP_ RPBSA_ M2rtTA _ Puro plasmid was constructed, then the recombinant plasmid and the “ Sleeping Beauty” plasmid expressing a transposase were cotransfected into HEK293T cells using lipofectamine. The inducible stable 293T (Tet1CD) cell line was selected using puromycin. Tet1 protein expression was detected via western blot, and the 5hmC content in the genome was detected using DNA Dot Blot. The effect of Tet1CD protein on the cell cycle and apoptosis was detected using flow cytometry, and the effect of Tet1CD protein on cell proliferation was detected by cell counting. Results The HEK293T (Tet1CD) cell line with inducible Tet1CD expression was successfully established. Western blot and DNA Dot Blot assays showed increased amounts of Tet1CD proteins and increased 5hmC content in inducible cells. Flow cytometry result demonstrated that Tet1CD expression promoted apoptosis (P< 0. 05) and increased the proportion of cells in G2 phase (P< 0. 05). Cell counts showed that Tet1CD overexpression inhibited cell proliferation (P< 0. 0001). Conclusions Tet1 protein may inhibit cell proliferation by promoting apoptosis and delaying the cell cycle.

Abstract:Objective To investigate effects of MD1 deficiency on the susceptibility of obese mice to ventricular arrhythmias and the mechanism involved. Methods Sixteen MD1-knockout (KO) mice and 16 male wild type (WT) mice were divided into WT normal group, MD1-ko^- normal group, WT high fat group and MD1-ko^- high fat group. The high fat groups were fed a high-fat diet (fat energy accounted for 60%) for 20 weeks, and the normal groups were fed a normal diet (fat energy accounted for 10%). The body weight (BW), heart weight ( HW), blood glucose level, total cholesterol, triglyceride and low-density lipoprotein ( LDL) cholesterol levels, and HW/ BW ratio of the mice were measured. RR spacing, PR spacing, QRS duration, QTc interval, LVEDd, LVEDs, LVFS, and LVEF were measured by electrocardiography and echocardiography. Western blot was used to analyze TLR4, MyD88, P-CAMKII, Collagen I, Collagen III and TGF-β1 protein levels. Cardiac hypertrophy and fibrosis were observed in mice by HE staining and PSR staining. Results The body index, blood glucose, total cholesterol, triglyceride, LDL cholesterol, QTc interval, LVEDd, LVEDs, LVFS, and LVEF were significantly higher in the MD1-ko^- high fat group than in the WT high fat group (P < 0. 05). Furthermore, the action potential duration ( APD20, APD50, APD90) of the MD1-ko^- high fat group was significantly higher than that of the WT high fat group (P <0. 05), and he APD alternating threshold of the MD1-ko^- high fat group was significantly higher than that of the WT high fat group (P< 0. 05). The arrhythmia ratio of the MD1-ko^- high fat group was significantly higher than that of the WT high fat group (P < 0. 05). The expression of TLR4, MyD88, and P- CAMKII in the MD1-ko^- high fat group was significantly higher than that in the WT high fat group (P < 0. 05). The HW/ BW ratio in the MD1-ko^- high fat group (7. 59 ± 0. 78) was significantly higher than that in the WT high fat group (6. 07 ± 0. 58; P < 0. 05). The Gross hearts of the MD1-ko^- high fat group (381. 23 ± 35. 76) μm²was significantly higher than that of the WT high fat group (190. 15 ± 25. 23) μm² ; P < 0. 05). Finally, the protein expression of Collagen I, Collagen III, and TGF-β1 in the MD1-ko^- high fat group was significantly higher than that in the WT high fat group (P< 0. 05). Conclusions MD1 deficiency increases the susceptibility to high-fat-diet-induced arrhythmias, mainly because of enhanced activation of the TLR4 / MyD88 signaling pathway, leading to left ventricular hypertrophy and fibrosis, which increases the expression of TLR4 / MyD88 signaling pathway-related proteins.

Abstract:Objective To calculate and analyze blood pressure data and histamine sensitivity of domestic cats on examination of depressor substance, and to provide a reference for the use of cats in experiments. Methods Blood pressure and histamine sensitivity were measured in domestic cats using the test method described in the Appendix of China Pharmacopoeia ( edition 2015) after analyzing the combined anesthesia data. Results Blood pressure and histamine sensitivity measurements were not correlated with body weight, whereas histamine sensitivity was correlated with blood pressure in this study. Furthermore, cats with blood pressure within the range of 16 ~ 18 kPa had a greater capacity for repeat examinations of depressor substances. Blood pressure and histamine sensitivity measurements did not decrease during the first six rounds of experiments. While blood pressure decreased significantly on the seventh and eighth rounds (P < 0. 05), and histamine sensitivity decreased on the eighth round (P < 0. 05), compared with data from the first round, the result still met the requirements of China Pharmacopoeia. Conclusions Blood pressure and histamine sensitivity measurements in cats were relatively stable, suggesting that examinations of depressor substances can be performed repeatedly in domestic cats.

Abstract:Objective Proteus mirabilis was isolated from the intestines of introduced cynomolgus monkeys and identified and tested for drug sensitivity to provide theoretical and technical support for detecting microorganisms from introduced experimental monkeys and to conduct breeding management and disease control. Methods Fecal samples were collected from cynomolgus monkeys via anal swabs, and bacterial isolation and culturing, biochemical identification, drug sensitivity tests, and mouse pathogenicity tests were performed. Results Suspicious bacteria were isolated from 8 of 60 cynomolgus monkeys and identified as Proteus mirabilis, indicating a 13. 3% infection rate of Proteus mirabilis in these monkeys. Susceptibility tests of this bacterium showed that it was highly sensitive to ceftriaxone, cefotaxime, cefoperazone, cefazolin, and penicillin; moderately sensitive to amikacin, gentamicin, ampicillin sodium, and furantoin; sensitive to norfloxacin and tetracycline; lowly sensitive to bacitracin, ciprofloxacin, kanamycin, and lincomycin; and resistant to furazolidone. Mouse pathogenicity tests showed that the isolated strains were highly pathogenic to mice. Conclusions The Proteus mirabilis infection rate was relatively high in cynomolgus monkeys. This bacterium was sensitive to cephalosporin antibiotics; therefore, cephalosporins can be used to clinically treat infections with this bacterium. Proteus mirabilis is highly pathogenic and potentially harmful to cynomolgus monkey populations and should be considered in their normal breeding management.

Abstract:Objective To investigate the role of the commercial Youcha powder in preventing blood lipid elevation caused by high-fat diet (HFD). Methods Forty male SD rats were randomly divided into five groups: the Control Group was given normal diet, and the HFD Group, HFD+L Group, HFD+M Group and HFD+H Group were given high-fat diet. The first two groups were given water, and the HFD+L, HFD+M and HFD+H groups were given low-dose, medium-dose and high-dose Youcha, respectively, by gavage. After 4 weeks, the gavage was stopped for 2 weeks. Blood was taken from the submandibular vein from 2nd to 6st weekend to determine serum lipid levels. Results From 2nd to 6st weekend the levels of total cholesterol (TC) in the HFD Group were significantly higher than those in the Control Group (P< 0. 001,P< 0. 05, P < 0. 01,P< 0. 05,P< 0. 05, respectively). From 2nd to 4st weekend, the TC in the HFD + L group was significantly lower than that in the HFD Group (P< 0. 05,P< 0. 05,P< 0. 01, respectively). The TG in the HFD+H Group at 3rd and 4st weekend were significantly lower than that in the HFD Group (P< 0. 05,P< 0. 01, respectively). Conclusions Youcha powder inhibited the increase in blood lipids caused by high-fat diet.

Abstract:Objective To examine the sensitivity of clear-cell renal carcinoma cells ( ccRCCs) using patients- derived cells (PDCs) to molecular-targeted drugs in patient-derived tumor xenograft (PDX) models.Method PDCs from ccRCCs were subcutaneously inoculated into nude mice to establish a tumor model. The molecular-targeting drugs, sunitinib, sorafenib, lenvatinib, regorafenib, apatinib and anlotinib, were orally administered to examine the inhibitory effect of the drug on the subcutaneous formation of PDC-derived ccRCCs in nude mice.Cell and tumor tissue samples were analyzed via qPCR. The targets of the molecular-targeting drugs (i.e., receptor tyrosine protein kinases, such as VEGFR, and protein kinases of the MAPK signaling pathway such as ERK and AKT) were examined.Result Five lines of PDC- derived ccRCCs were successfully established, and the nude mice were injected with PDC-derived ccRCC to obtain a subcutaneous tumor model of kidney cancer in nude mice. Expression of the target molecules of these drugs decreased during in vitro PDC culturing. Moreover, expansion of these molecules in the tumor tissues was relatively stable. The inhibitory rates of the molecular-targeting agents differed for each drug, and the antitumor activity of lenvatinib was stronger than that of several of the other drugs.Conclusions PDC-derived ccRCCs can be used to establish a mouse model of kidney cancer and test the sensitivity of renal cancer cells to molecular-targeted drugs. These models can provide a rational and experimental basis for relevant clinical diagnosis and treatment.

Abstract:Objective To investigate the effect of astragaloside on the TLR4-p38 MAPK signaling pathway in a newborn mouse acute lung injury model. Methods Seven-day-old SD newborn rats were used as experimental subjects. The model of acute lung injury in newborn rats was established by intraperitoneal lipopolysaccharide ( LPS) injection. Dexamethasone was used as a positive control. Each rat was injected intraperitoneally with 10 mL/ kg astragaloside. The lung lesions were observed 12 and 24 h after LPS injection. Furthermore, the mRNA and protein expression levels of TLR4 were determined, as well as the phosphorylation level of p38 protein. Finally, the changes in inflammatory factors TNF-α, IL-6 and IL-12 were analyzed. Results The model group showed pulmonary hemorrhage and a large number of collapsed alveoli at 12 h, and pulmonary edema and a significantly widened alveolar septum at 24 h after LPS injection. Compared with the model group, the lesions in the lung tissue of the astragaloside group were significantly alleviated. Additionally, Western blot result showed the levels of TLR4 (P< 0. 05) and p-p38 (P< 0. 001) were significantly lower in the astragaloside group than in the model group. The qPCR result showed that the level of TLR4 mRNA in the astragaloside group was significantly decreased compared with that in the model group (P< 0. 05). The inflammatory factors, TNF-α, IL-6 and IL- 12, were all decreased, and the relative IL-12 expression was significantly different from that in the model group (P< 0. 05). Conclusions Astragaloside plays an important role in alleviating the acute lung injury model of newborn rats by inhibiting the TLR4-p38 MAPK signaling pathway.

Abstract:Objective To introduce an improved method for intubating the jugular vein and for jugular vein intubation surgery in rats and to apply this method to a rat self-administration experimental study to evaluate the service life and application effect of this intubation design. Methods Twenty-nine male Sprague-Dawley rats who had completed food training were intubated with a modified cannula in the jugular vein. Self-administration models of propofol and cocaine were established, and elimination from the rats was analyzed at approximately 5 months postoperation. Results Both the propofol and cocaine self-administration models were successfully established. The main reasons for postoperational drug elimination were death of the rats, blocked tubes, and fluid leakage. In months 1-5 after surgery, the intubation-related elimination rates for each month were 7%, 7%, 10%, 31%, and 11%, respectively. With an acceptable monthly elimination rate of <10%, the service life of the intubation is expected to be at least 3 months, and some can be as long as 5 months or more. Rat deaths were related to severe liver and kidney damage, electrolyte disturbances and inflammation. Conclusions Improved intubation of rat jugular veins for drug self-administration has many advantages. The optimal service life of this intubation technique was up to 3 months, which meets the basic requirements of self-administration experiments. Therefore, this application effect is good and should be promoted.

Abstract:As an important vertebrate model organism, zebrafish are widely used in research fields such as developmental biology. With the rapid development of zebrafish research, experimental zebrafish housing must be standardized and should facilitate zebrafish husbandry. Here, we systematically discuss the construction of zebrafish facilities, including the automatic breeding system and the building design and renovation. The zebrafish breeding system uses modern industrial automation technology to provide a safe and suitable environment for zebrafish growth and breeding. Suitable zebrafish facilities should integrate the breeding system with the building structure.

Abstract:Regional anesthesia is important in clinical anesthesia work, and with the wide application of visualization tools such as ultrasound, regional anesthesia is increasingly used by clinical anesthesiologists. However, in the basic and clinical research of regional anesthesia, the corresponding research has been hindered by the lack of reliable and stable animal models of regional anesthesia. In recent years, with the assistance of new visualization tools, great progress has been made in the establishment of regional anesthesia animal models. In this review, the types and establishment method of regional anesthesia animal models that promoted the development of regional anesthesia were reviewed, and the future prospects were discussed.

Abstract:Laboratory fish are small in size and caliber, and their microdiet has limited uses because of poor stability, food inductivity, digestibility and water pollution. In contrast, live food is appropriately sized, rich in nutrition, and does not pollute the water quality; thus, it has long been recognized as the main feed for laboratory fish. Herein, we review the biological characteristics; nutrient composition; possible pathogens, toxins, and harmful substances; and quality control indexes of live food, such as Paramecium caudatum, Brachionus plicatilis and Artemia salina, which are commonly consumed by laboratory fish, to provide a quality control reference for laboratory fish feed.

Abstract:Thrombotic diseases are a major threat to human health, ranking first in causes of death worldwide. Endothelial nitric oxide synthetase ( eNOS) is the main source of nitric oxide, an important protective factor of the cardiovascular system. As an important bioactive substance protecting the vascular wall, eNOS participates in the pathophysiological processes of thrombotic diseases such as acute coronary syndrome, atrial fibrillation, stroke, pulmonary thromboembolism and deep vein thrombosis. This paper reviews the structure and physiological functions of eNOS, its mechanisms, and research progress in thrombotic diseases.