Abstract: Objective To construct a lentiviral vector for RNA interference of the Sirt3 gene and to establish a Sirt3 knockdown human neuroblastoma SH-SY5Y cell line. Methods According to the Sirt3 nucleotide sequence archived in the GenBank database, four siRNAs targeting Sirt3 and one negative control were designed, and cloned into a linear vector containing the green fluorescent protein ( GFP ) gene to produce four recombinant lentivirus plasmids. The recombinant plasmids and helper plasmids were transfected into 293T cells, and the titer of the virus was determined. SH- SY5Y cells were infected with the constructed lentivirus, and the silencing effect on Sirt3 was accessed by real-time PCR and Western blot. The lentivirus-infected cells were screened for the most significant Sirt3 knockdown. Results Recombinant lentiviral vectors expressing siRNAs targeting Sirt3 were successfully constructed and confirmed by DNA sequencing. The viral titers of the recombinant lentivirus were as follows: LV-Sirt3-RNAi-1 8×10⁸ TU/ mL, LV-Sirt3- RNAi-2 3×10⁸ TU/ mL, LV-Sirt3-RNAi-3 8× 10⁸ TU/ mL, and LV-Sirt3-RNAi-4 8× 10⁸ TU/ mL. The levels of Sirt3 mRNA and Sirt3 protein in the LV-Sirt3-RNAi-3 group were significantly less than those in the negative control group and the blank control group ( P < 0. 001, P < 0. 001). Conclusions A lentivirus vector for RNA interference of Sirt3 was successfully constructed and SH-SY5Y cell lines with Sirt3 gene knockdown were selected, which will be useful for future research of Sirt3 function in Parkinson’s disease cell models.

Abstract: Objective To explore alleviation of the inflammatory response induced by acute lung injury in sepsis rats by down-regulation of miR-128-3p and effects on lung tissue morphology. Methods Sixty healthy 3-week-old SD rats were divided into a sham-operated group, a sepsis model group, a blank transfection group and an miR-128-3p inhibitor group by a random grouping method with 15 rats in each group. Sepsis models were prepared by cecal ligation and puncture. In the blank transfection group, blank plasmids were constructed and were transfected into rats. In the miR-128-3p inhibitor group, lentiviral plasmids were constructed and were transfected into rats. The expression of miR-128-3p was detected by RT-PCR to determine the success of interference. Resting ventilation, changes in airway resistance and lung volume were determined in each group. The contents of serum IL-6, TNF-α and iNOS in peripheral blood were detected by ELISA. Lung tissues and pathological injury were observed by hematoxylin & eosin ( HE) staining. The expression of Caspase-3 and Caspase-9 in lung tissue was detected by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) staining and western blotting. The expression of TGF-β and α-SMA was detected by Masson 's trichrome staining and western blotting. Results HE staining showed that cell distribution was even in the miR-128-3p inhibitor group, with few necrotic cells. MASSON staining showed that there was severe fibrosis and damage of lung tissues in sepsis model group, and degree of lung tissue fibrosis in miR-128-3p silencing group was improved. Compared with the sham-operated group, expression of miR-128-3p, airway resistance, levels of IL-6, iNOS and TNF-α, apoptosis rate of lung tissue cells, relative expression of cleared cas3 / Caspase-3, cleared cas9 / Caspase-9, and TGF-β and α-SMA protein levels were significantly increased (P< 0.05), while resting ventilation, changes in airway resistance and lung volume were significantly decreased in the sepsis model group (P<0.05). Compared with the blank transfection group, expression of miR-128-3p, airway resistance, levels of IL-6, iNOS and TNF-α, apoptosis rate of lung tissue cells, relative expression of cleared cas3 / Caspase-3, cleared cas9 / Caspase-9, and TGF-β and α-SMA protein levels were significantly decreased ( P < 0. 05), while resting ventilation, changes in airway resistance and lung volume were significantly increased in the miR-128-3p inhibitor group (P<0.05). Conclusions Down-regulation of miR-128-3p can alleviate acute lung injury in sepsis rats and the mechanism of action may be related to a reduced inflammation response and inhibited apoptosis of lung tissue cells.

Abstract: Objective To investigate the effects of agmatine on Propofol-induced neurotoxicity in newborn rats. Methods Rats were divided into five groups for follow-up experiments: Control, Propofol, Propofol + 1 mg / kg Agmatine, Propofol + 2. 5 mg / kg Agmatine, and Propofol + 5 mg / kg Agmatine. The number of errors was detected in a platform experiment, while brain moisture content and brain index were determined by a 2, 3, 5-triphenyl tetrazolium chloride method. The Longa method was used to evaluate neural functional defect and postural reflex scores. HE staining was used to detect the degree of pathological damage in the hippocampus. Apoptosis of hippocampal tissue was detected by Nissl staining. Western blot was used to detect protein expression levels of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), Bax, Bcl-2, Nrf2, p-Nrf2, and heme oxygenase 1 (HO-1). Results Compared with the Control group, the Propofol group exhibited significant increases in the number of platform errors (P<0. 05), brain water content and cerebral index (P<0. 05), neural function defect scores, and posture reflex scores (P< 0. 05), as well as obvious pathological damage. Moreover, the Propofol group exhibited significantly reduced expression of BDNF, NGF, and HO-1 proteins (P<0. 05), p-Nrf2 / Nrf2 ratio (P<0. 05), and numbers of Nissl bodies (P<0. 05), but a significantly higher Bax / Bcl-2 ratio (P<0. 05). Compared with the Propofol group, Propofol+Agmatine groups (2. 5 and 5 mg / kg) exhibited significantly decreased numbers of platform errors ( P< 0. 05), brain water content and cerebral index ( P< 0. 05), neurologic deficit scores (P<0. 05), and posture reflex scores (P<0. 05). Moreover, the degree of pathological damage was significantly improved in Propofol+Agmatine groups, which exhibited significantly increased BDNF and NGF protein expression (P<0. 05), and numbers of Nissl bodies (P<0. 05), and a significantly decreased Bax / Bcl-2 ratio (P<0. 05;. p-Nrf2 / Nrf2 ratio and HO-1 protein expression were also significantly increased ( P<<0.05). Conclusions Agmatine attenuates Propofol-induced neurotoxicity in newborn rats by regulating the Nrf2 / HO-1 signaling pathway.

Abstract: Objective To knock out the high-mobility group AT-hook 2 (HMGA2) gene using the CRISPR/ Cas9 system in HepG2 cells and investigate the effect of HMGA2 knockout on the growth, proliferation, migration, and invasion of hepatoma carcinoma cells. Methods The human HMGA2 gene sequence was obtained from GenBank. Two sgRNAs were designed for each of the first and second exons of the HMGA2 gene using online sgRNA design software. The recombinant sgRNA vectors sgE1 and sgE2 were constructed and transfected into HepG2 cells to obtain the HMGA2^{- / -} cell line. The effects of HMGA2 knockout on the growth and proliferation of HepG2 cells were investigated via CCK8 and clone formation assays, while the migration and invasion abilities were measured using Transwell assays. Results Compared with wild-type HepG2 cells, the knockout cell line showed reduced proliferation, clone formation (121. 83 ± 21. 68 vs 59. 50 ± 20. 68, P< 0. 01), invasion (359. 67 ± 32. 53 vs 245. 61 ± 24. 23,P< 0. 05), and migration ( 251. 33 ± 43. 43 vs 47. 00 ± 10. 00, P< 0. 01). Conclusions HMGA2 knockout in hepatoma carcinoma cells inhibited both cellular proliferation and tumor metastasis in vitro. Therefore, HMGA2 may constitute a target for hepatoma gene therapy.

Abstract: Objective To investigate the effects of curcumin on microRNA-125b (miR-125b) expression and cell damage in high glucose-induced human retinal pigment epithelial (RPE) cells. Methods ARPE-19 cells were cultured in vitro and divided into a control group, a high glucose group, a high glucose + curcumin group, a high glucose + miR-125b inhibitor group and a high glucose + inhibitor-negative control (NC) group. MTT assays were used to detect the activity of ARPE-19 cells. Annexin V-FITC/ PI double staining was used to detect apoptosis of ARPE-19 cells. A 2′ - 7′ dichlorofluorescin diacetate (DCFH-DA) probe loading method was used to detect the level of reactive oxygen species (ROS). An enzyme-linked immunosorbent assay (ELISA) was used to detect the level of superoxide dismutase (SOD) and RT-qPCR was used to detect the expression of miR-125b in cells. Results MTT assays showed that curcumin significantly improved the activity of ARPE-19 cells in a high glucose environment. Curcumin significantly reduced the rate of ARPE-19 cell apoptosis and ROS production in a high glucose environment, and increased the expression of SOD. RT-qPCR showed that curcumin significantly reduced the expression level of miR-125b. Conclusions Curcumin may reduce ROS production and increase SOD expression by down-regulating miR-125b expression. This would improve the antioxidant capacity of ARPE-19 cells and protect them from high glucose-induced cell damage.

Abstract: Objective To investigate the effects of vacuolar protein sorting 4B ( VPS4B) on odontogenic differentiation and Notch pathway signaling in human dental pulp stem cells ( hDPSCs). Methods hDPSCs was divided into normal, mineralized, negative control, and VPS4B-siRNA groups. The normal group was grown in normal culture, without any additional treatment. The remaining groups underwent treatment with mineralized liquid ( 0. 785 g / L dexamethasone, 0. 05 g / L vitamins, 2. 16 g / L sodium β-glycerophosphate); the negative control and VPS4B-siRNA groups underwent transfection of negative control siRNA and VPS4B-siRNA, respectively, using Lipofectamine 2000. The CCK8 assay was used to detect cell proliferation. The protein levels of VPS4B, Notch intracellular domain (NICD), and hairy and enhancer of split-related with YRPW motif1 (Hey1) were detected by Western blotting. Cell mineralization was observed by alizarin red staining. Cell calcium concentrations were determined using a calcium detection kit. The mRNA levels of Runx2, osteopontin (OPN), and dentin sialophosphoprotein (DSPP) were detected by real-time fluorescence quantitative PCR. Results In the normal group, the alizarin red staining area was light and small. The mineralized and negative control groups exhibited a darker and larger staining area, while the VPS4B-siRNA group exhibited a lighter and smaller staining area. Compared with the normal group, the levels of OD₄₅₀ , NICD, and Hey1 proteins were reduced in the mineralized and negative control groups (all P< 0. 05), while the relative number of mineralized nodules, level of VPS4B protein, concentration of calcium, and levels of Runx2, OPN, and DSPP mRNA were enhanced ( all P< 0. 05). The relative number of mineralized nodules and levels of OPN and DSPP mRNA were enhanced in the VPS4B-siRNA group (all P< 0. 05), while the levels of OD₄₅₀ , NICD, and Hey1 proteins were reduced ( all P< 0. 05). Compared with the mineralized and negative control groups, NICD and Hey1 protein levels were enhanced in the VPS4B-siRNA group (both P< 0. 05), while the levels of OD₄₅₀ and VPS4B protein, the relative number of mineralized nodules, concentration of calcium, and levels of Runx2, OPN, and DSPP mRNA were reduced ( all P< 0. 05). Conclusions Interference with VPS4B activity can inhibit odontogenic differentiation in hDPSCs, presumably through modulation of the Notch pathway.

Abstract: Objective The genetic structure composition of the ddY mouse population, which was bred in isolation for a long time, was compared with that of the newly introduced ddY mouse population by microsatellite analysis. Methods Thirty-six microsatellite loci were analyzed. Thirty-eight DNA samples were randomly collected from each of the two groups. The DNA samples were then amplified by PCR and the PCR products were sequenced. PopGen1. 32 and Littleprogram0. 6 were used to process the data. Results The average allele number of group A was 2. 4839, the average heterozygosity was 0. 3376, and the average polymorphism information content was 0. 2875. Group B had an average allele number of 3. 4839, average heterozygosity of 0. 4806, and an average polymorphism information content of 0. 4088. Conclusions The genetic structure of the two laboratory mouse populations was different, and closed breeding result ed in genetic structure changes in the populations. It is necessary to monitor the genetic structure of closed laboratory animal populations regularly to ensure the genetic quality of populations.

Abstract: Objective To investigate the effect of pomegranate blood-enriching syrup on anemia in Wistar rats. Methods An anemia model was established in rats by feeding a low-iron diet. Different doses of pomegranate blood- enriching syrup were then given for 20 consecutive days. Blood samples were taken on the 10th day for routine blood analysis. On the 20th day, rats were euthanized and blood and organs were collected. Serum erythropoietin (EPO) levels and the number of nucleated cells were determined. Nucleic acid and protein levels of stem cell factor ( SCF) and granulocyte-macrophage colony-stimulating factor ( GM-CSF) in bone marrow mesenchymal stem cells were detected. Results The clinical symptoms of the different pomegranate blood-enriching syrup dose groups were relieved compared with those of the model group. Mortality was reduced and the number of peripheral blood cells increased. Serum EPO levels were decreased. Compared with the model group, the number of nucleated cells in the bone marrow and the levels of SCF and GM-CSF in the medium dose group were increased. Conclusions Pomegranate blood-enriching syrup has a clear therapeutic effect on anemia in Wistar rats.

Abstract: Objective Gender differences in immune cell types in peripheral blood and spleen were compared between aged and young Sprague-Dawley rats. Methods Twelve-month and 2-month-old rats were used. Rats in each age group were half male and half female. Classification and counts of peripheral blood cells, organ index, peripheral blood and spleen immune cell typing, and P16 expression in spleen T cells were determined and the result compared with respect to age and gender differences. Results The peripheral blood leucocyte count was decreased in the aged female rats, and there was a gender difference. The erythrocyte count was increased in both aged male rats and aged female rats, but there was a difference between them. The neutrophil percentage was increased and the lymphocyte percentage was decreased in the aged male and female rats. The eosinophil percentage was increased in the aged male and female rats. The monocyte percentage in the aged female rats was lower than that in the aged male rats. No significant changes in other peripheral blood indices were found. The thymus index of the aged female rats was decreased and was lower than that of the aged male rats. The spleen index of the aged male rats was increased and was higher than that of the aged female rats. Peripheral blood immunophenotyping indicated that the percentages of T helper cells, regulatory T cells and cytotoxic T cells were increased in the aged female rats. The percentage of B cells was decreased in the aged female rats. The percentage of natural killer cells was increased in the aged male rats. Splenic lymphocyte typing showed that numbers of CD3+ cells increased and CD45RA+ cells decreased in aged female rats. P16 expression in splenic T cells increased in all aged rats, and no gender difference was observed. Conclusions No gender differences were found in the various indices among young rats. Some indices showed a gender difference in aged rats, including peripheral blood leucocyte count, erythrocyte count, neutrophil percentage, monocyte percentage, thymus index, spleen index, and immune cell typing for regulatory T cells, cytotoxic T cells, natural killer cells, B cells and CD3-positive cells. These result indicate altered immune system homeostasis in aged rats, leading to gender differences in some indices in aged rats. The present result provided basic data for studying the impact of gender on aging-related diseases and animal models of aging.

Abstract: Objective To test the stability of serum samples in proficiency tests. Methods In accordance with CNAS-CL04(CNAS: China National Accreditation Service for Conformity Assessment), serum samples of inbred mice were tested for storage stability, acceleration stability, transport stability, and final stability. Results The samples were stable at -20℃ or 4℃ for 50 days. The samples were stable at 37℃ or room temperature for 27 days. The samples were stable during transportation. All samples passed the final stability test. Conclusions The collected serum samples met the requirements for proficiency tests. Serum samples of esterase-1 were adequate for proficiency testing.

Abstract: Objective To evaluate the anesthetic effect and safety of premedication midazolam combined with butorphanol on isoflurane-anesthetized mice. Methods Seventy five mice were randomly divided into three groups of 25 mice: control group(receiving isoflurane anesthesia only), observation group(receiving premedication midazolam combined with butorphanol with isoflurane anesthesia)and blank group(receiving no anesthesia). The indices of heart rate(HR) and oxygen saturation(SpO₂ )were assessed at induction and every 5 min post-induction for 1 h. The serum levels of glucose (Glu)and cortisol(Cor)were measured and compared pre- and post-induction. The indices of induction and recovery time, and analgesic and sedative time were measured and compared among the three groups. Adverse reactions during induction were also recorded. Results Compared with the control group, the indices of induction time, sedation onset time and pain- loss time were shorter in the observation group(P<0. 05), but the recovery time was no different(P>0. 05). The HR index in the blank group was the highest among the three groups(P<0. 05). The HR index of the control group was higher than that of the observation group during 5~ 35 min post-induction(P<0. 05), and the HR index of the control group was lower than that of the observation group during 35~ 60 min post-induction(P<0. 05). The SpO₂ index of the control group was the lowest among the three groups after 1 h post-induction ( P < 0. 05), and the SpO₂ index was not different between the observation and blank groups(P>0. 05). The HR variation tendency was different among the three groups(F= 15. 215, P= 0. 000). The SpO₂ variation tendency was not different among the three groups(F= 1. 015, P= 0. 180). The serum levels of Glu and Cor were not different among the three groups pre-anesthesia(all P>0. 05). The serum levels of Glu and Cor in the control group were the highest among the three groups at mid-anesthesia(all P<0. 05), but no difference was found between the blank and observation groups at mid-anesthesia(all P>0. 05). The serum levels of Glu and Cor at mid-anesthesia were higher than at pre-anesthesia in the control group ( P< 0. 05), but the serum levels of Glu and Cor in the blank and observation groups were not different between pre-anesthesia and mid-anesthesia ( P> 0. 05). The incidences of adverse reactions were not different between control and observation groups(P= 0. 626). Conclusions The anesthetic effects of premedication midazolam combined with butorphanol on isoflurane-anesthetized mice are safe and reduce the degree of respiratory inhibition.

Abstract: Objective To observe the expression of c-Myc, RhoA and YAP proteins in the kidney and to determine their relationship with tubulointerstitial fibrosis in adenine-induced chronic kidney disease. Methods Thirty rats were randomly divided into a control (Con) group and a CKD (adenine-induced chronic kidney disease rat model) group. Rats were euthanized at weeks 4, 6, and 8. The levels of renal function and 24-hour urine protein in the two groups were determined. Hematoxylin & Eosin (HE) staining was used to observe pathological changes in the kidney and to score the degree of renal interstitial injury. Interstitial fibers of renal tubules were observed by Sirius red staining and the degree of fibrosis was determined. Immunohistochemical staining was used to observe the expression of c-Myc, RhoA and YAP proteins in renal tissue. RT-PCR was used to detect the mRNA levels of c-Myc, RhoA, YAP, Vimentin, Col1, and α- SMA. The relationship between the expression of c-Myc, RhoA, and YAP and renal tubular interstitial fibrosis was analyzed. Results Compared with the Con group, the blood urea nitrogen(BUN), serum creatinine( Scr), cystatin C (Cysc), and the 24-hour urine protein level of the CKD group were significantly increased ( (P< 0. 05). HE staining showed that the renal tubules were significantly expanded, and Sirius red staining showed a significant increase in collagen fibers. Immunohistochemistry showed that the expression of c-Myc, RhoA, YAP, Col1, and αSMA proteins in the CKD group was significantly higher than that in the Con group (P<0. 05). The levels of c-Myc, RhoA, and YAP mRNA were positively correlated with Col1 mRNA (r= 0. 927 P= 0. 001; r= 0. 988 P= 0. 001;r= 0. 745 P= 0. 013, respectively) and α SMA mRNA levels (r= 0. 988; P= 0. 001; r= 0. 818; P= 0. 004;r= 0. 770; P= 0. 009, respectively). Additionally, a significant positive correlation was found between c-Myc and RhoA mRNA (r= 0. 721 P= 0. 019), and RhoA and YAP mRNA (r=0. 661 P= 0. 038). Conclusions In a rat adenine-induced renal tubular interstitial fibrosis model, c-Myc, RhoA and YAP proteins were highly expressed and were closely related to the degree of renal tubular interstitial fibrosis.

Abstract: Objective To investigate the mechanism by which miR-335-5P regulates the transforming factor-β (TGF-β) pathway in promoting islet β cell secretion and enhancing insulin resistance in gestational diabetes mellitus (GDM) mice. Methods Specific pathogen free grade Balb / c mice were reared in cages at a female to male ratio of 2 ∶1. Ten normal pregnant mice were assigned to the blank group, and 48 mice were prepared as GDM model mice and divided into six groups for different drug treatments—groups A, B, C, D, E and F—with eight mice in each group. After drug treatment, fasting glucose (FBG) and fasting insulin (FIRI) were detected by the oral glucose tolerance test (OGTT), and the insulin resistance index (IRI) was calculated. The hyperinsulinemic-euglycemic clamp was used to determine the glucose infusion rate (GIR) and to estimate pancreatic islet β cell function. The levels of miR-335-5P and key islet β cell TGF-β pathway factor mRNAs and proteins were determined by RT-qPCR, western blot and ELISA analysis. Results After drug treatment, FBG, FIRI and IRI in groups D and F were decreased and were lower than respective values of groups C and E (P<0. 05). GIR, first-phase insulin and maximum insulin in groups D and F were increased and were higher than respective values of groups C and E (P<0. 05). The levels of miR-335-5P, TGF-β and c-Myc in groups D and F were lower than those of groups A, B, C and E (P<0. 05). When miR-335-5P expression was detected in islet β cells, TGF-β and c-Myc were expressed at different levels. TGF-β was expressed in the cytoplasm of islet β cells, and the expression of TGF-β in groups D and F was higher than that in groups A, B, C and E (P<0. 05). Conclusions miR-335-5P can up- regulate c-Myc expression in GDM mice, activate the TGF-β pathway, enhance insulin resistance and inhibit islet β-cell secretion.

Abstract: Objective This study focused on the effects of different enrichment designs on behavior and cortin levels in male SD rats. Methods Forty-five 6-week-old male specific pathogen free SD rats were randomly divided into four groups. The single cage group had nine rats and the other three groups each had 12 rats. The novelty group was housed in a composite environmental enrichment scheme composed of nesting material, shelter, grinding rods, and a new scheme was provided every week. The environmental enrichment group provided one kind of material every week. The control group and the single cage group did not receive enrichment. The experiment lasted 9 weeks. Behavior was sampled using instantaneous scanning sampling method. Serum and urine adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels were measured. Results Novelty group rats exhibited more exploring behaviors than rats in the other groups. Rats of the single cage group increased their inactive behavior. No significant differences were found among groups in the serum levels of ACTH or CORT. However, in urine, the concentration of ACTH and CORT showed the following trend: novel grouP>single cage grouP>control grouP> environmental enrichment group. Conclusions Single cage housing inhibits the expression of behaviors in SD rats, which is detrimental to their welfare. However, the complex novel enrichment program increased positive behaviors, such as exploration behavior and also has an effect on adrenocortical hormones. In practice, simple and practical environmental enrichment materials and schemes should be selected according to specific housing conditions.

Abstract: Objective To observe the effect of liraglutide on the expression of glucose-6-phosphatase (G6pase) and phosphoenolpyruvate carboxylase (PEPCK) in the diabetic mouse liver and explore the underlying mechanisms of these effects. Methods Mice were divided into three groups: the normal control group ( male C57 mice, n= 5) received intraperitoneal injection of normal saline, the model control group ( male KK-Ay mice, n= 5) received intraperitoneal injection of normal saline, and the Liraglutide group (male KK-Ay mice+ drug intervention, n= 5) received intraperitoneal injection of liraglutide. All three groups of mice were raised in the same feeding environment. After 8 weeks of intervention, the blood glucose of mice was detected; the expression levels of the FOXO1, cryptochrome 1, the E3 ubiquitin ligase DDB1, G6pase, and PEPCK in the liver were detected by Western blotting. Results Compared with the model control group, blood glucose was significantly reduced in the intervention group. The expression levels of DDB1, FOXO1, G6Pase and PEPCK decreased in the intervention group, while the expression level of Cryptochrome 1 ( CRY1) increased. Conclusions Liraglutide can downregulate the expression of G6Pase and PEPCK, key gluconeogenesis enzymes, in the diabetic mouse liver; this may be related to downregulation of the E3 ubiquitin ligase DDB1.

Abstract: Objective To develop a rapid, high-sensitivity, and high-specificity assay based on TaqMan real- time fluorescent quantitative PCR (TaqMan real-time FQ-PCR) targeting the gag gene of simian-human immunodeficiency virus (SHIV) for quantitative measurement of simian immunodeficiency virus (SIV) and SHIV in plasma, peripheral blood mononuclear cells, and lymph nodes from rhesus macaques. Methods A 109-bp fragment of the SHIV_SF162p3n gag gene was amplified using reverse-transcription PCR and cloned into the pGEM? -T vector to construct the pGEM? -T-SHIV gag plasmid, which was used as a standard for the assay. Results TaqMan real-time FQ-PCR efficiently detected 10² – 10⁷ copies/ μL SIV/ SHIV gag mRNA with a correlation coefficient (R² ) of 0. 998 and slope of - 3. 304. The coefficient of variability was 0. 6%– 1. 1%. Conclusions This rapid assay exhibited both high sensitivity and high specificity. Thus, this method is suitable for studies involving SIV/ SHIV infection macaque models.

Abstract: Objective To investigate the effects of online teaching in a laboratory animal science course and provide guidance for future teaching activities. Methods A questionnaire survey was conducted among students at a College of Veterinary Medicine in Beijing, China, who had completed an online laboratory animal science course. Results The online laboratory animal science course met the basic teaching Objective established by the College of Veterinary Medicine. However, online teaching alone lacks a practical component; a combined approach using both online and on-site teaching method can provide more robust result. Conclusions These result provide some guidance for the future development of laboratory animal science courses. However, further efforts are needed to more comprehensively establish the teaching model of the course, explore additional teaching method , and improve the quality of instruction.

Abstract:Cardiovascular diseases constitute the largest burden to health worldwide. Ischemic heart disease seriously harms the physical and mental health of sufferers and its morbidity and mortality continue to increase. Coronary artery bypass grafting is a treatment for ischemic heart disease that can effectively rebuild the blood flow of the heart, but it has the problem of an insufficient long-term patency rate for venous grafts. Stable animal models can provide tools to investigate venous bypass graft diseases. This article summarizes the establishment and application of animal models of venous bypass graft disease to stimulate new ideas for research.

Abstract:Intrauterine infection can be caused by a variety of pathogens invading into the uterine cavity during pregnancy and can cause abnormal pregnancy, neonatal diseases and other adverse outcomes for mothers and infants. However, its pathogenesis is not yet clear, the effects of treatments are poor, and the prognosis is not good. Therefore, establishment of a stable and reliable animal model of intrauterine infection will facilitate further research into its pathogenesis, mechanisms of drug action, and prevention, which is of great significance for reducing adverse outcomes of mothers and infants and to improve the prognosis. At present, the animal models of intrauterine infection established in China and elsewhere mainly include Escherichia coli or lipopolysaccharide-induced intrauterine infection, Ureaplasma urealyticum intrauterine infection, Candida intrauterine infection, and cytomegalovirus intrauterine infection. This article mainly reviews the selection of experimental animals, the experimental design of different modeling method and their advantages and disadvantages.

Abstract:Ferroptosis is a recently discovered type of programmed cell death and accumulating evidence suggests that it plays an important role in respiratory diseases through a variety of mechanisms, such as iron-dependent lipid peroxidation and reactive oxygen species ( ROS) production; its role in the occurrence and development of respiratory diseases cannot be ignored. This review summarizes current knowledge of the role and mechanism of ferroptosis in the development and progression of respiratory diseases, and provides a theoretical basis for further research.