Abstract:Objective To establish the common iliac vein stenosis model in miniature pigs. Next, to observe the influence and evaluate the safety of simulated drug?coated balloons (DCB) on the stenosis segment of the common iliac vein. Methods Twenty young and healthy male miniature pigs were selected for intravenous general anesthesia. Under anesthesia, the animal model was established by oversized balloon dilatation. After the animal model was successfully established, the miniature pigs were randomly divided into an experimental group and control group. Stenosis segment veins in miniature pigs of the experimental group were occluded by two balloons and perfused with paclitaxel. These steps were not performed in miniature pigs from the control group. Venous blood specimens were taken before and two weeks after operation. Venography was performed and pathological sections of stenotic segment veins observed before and two months after the operation. Hepatorenal function index, diameter of the common iliac vein, and histological changes of the venous wall were compared between both groups. Results Seventeen miniature pigs survived to the end of the experiment. Two weeks after the operation, there were no significant differences ( P > 0.05) between the experimental group ( n = 9) and control group ( n = 8) regarding alanine aminotransferase, aspartate aminotransferase, creatine, and blood urea nitrogen values. However, two months after the operation, lumen diameter was significantly different ( P < 0.05) between the experimental group [(3.74 ± 1.24) mm] and control group [(2.25 ± 1.41) mm]. Compared with the untreated contralateral common iliac vein, the intima and media of the target vein were thicker in both the experimental group and control group. Indeed, this was more obvious in the control group than experimental group. Conclusions The common iliac vein stenosis model can be established in miniature pigs by oversized balloon dilatation. Local perfusion with paclitaxel has little effect on hepatorenal function of miniature pigs. Using this approach, simulated DCB can reduce the degree of common iliac vein stenosis in miniature pigs.

Abstract:Objective To observe the process of SIVmac239 adsorbing onto and infecting target cell line CEMx174 and revealing the basis for the virus entering target cells. Methods CEMx174 cells were infected by the SIVmac239 virus. Culture supernatants and cells were collected at various time points. The viral load in culture supernatants was detected by quantitative real?time PCR. Expression of intracellular viral protein p27 was detected by indirect immunofluorescence, intracellular staining, and Western blotting. Results CEMx174 cells absorbed the virus and were infected. Virus particles were detected on the cell membrane at 0.5 h. At 12 h, the level of viral RNA in the culture supernatant was reduced. Then up to 96 h, the expression of SIV p27 in the cytoplasm was increased, and the level of viral RNA in the culture supernatants increased continually. Conclusions When the virus infects its target cells, it is first adsorbed through virus?associated receptors and then enters the cell.

Abstract:Objective To observe the effects of the lactose content in feed, the time duration of chronic restraint stress and the duration of modeling on the diarrhea?predominant of irritable bowel syndrome (IBS?D) model rats, and to select a preferred method for establishing the IBS?D rat model. Methods Rats were randomly divided into a normal control group, model group A (30% lactose + bound for 1 h), model group B (30% lactose + bound for 1.5 h), model group C (45% lactose + bound for 0.5 h), and model group D (45% lactose + bound for 1 h). The visceral sensitivity was evaluated with the pain threshold at the time of abdominal wall retraction on the 7th, 10th, and 14th days after modeling. The diarrhea index was used to evaluate the extent of diarrhea. A multi?channel physiological signal acquisition processing system was used to record the wave in jejunum smooth muscle movement at 1 h after feeding. Wave amplitude, frequency analysis, and movement calculation were used to evaluate gastrointestinal dynamics. Results The capacity threshold in each model rat at 7, 10, and 14 days was significantly lower than that in the normal control group. Diarrhea indexes at 7, 10, and 14 days were obviously increased compared with the normal control group. Model group D had less significant abnormalities in rhythmic movement of smooth muscle. Conclusions When lactose in the feed is > 30% with a time duration of chronic restraint stress for more than 1 h, gastrointestinal motility abnormalities and visceral high sensitive state of typical IBS?D occur in the model rats at 7 -14 days after modeling.

Abstract:Objective To study the molecular mechanism of ligustilide (LIG) in HUVEC injury induced by ox?LDL. Methods HUVECs were randomly divided into five groups: control group (untreated cells), model group (cells treated with 100 μmol/ L ox?LDL for 24 h), and LIG groups (cells pre?treated with 10, 20, or 40 μmol/ L LIG for 2 h and then exposed to 100 μmol/ L ox?LDL for 24 h). Cell viability was measured by CCK8 assays. Levels of inflammatory cytokines TNF?α and IL?6, and ICAM?1 and VCAM?1 were assayed by ELISAs. mRNA levels of TNF?α, IL?6, ICAM?1, VCAM?1, and miR?181b were determined by qRT?PCR and the protein level of NF?κB by Western blotting. Results Compared with the control group, ox?LDL induced apoptosis and inflammatory cytokine secretion in HUVECs ( P <0.01). LIG protected cells and reduced apoptosis. Moreover, LIG reduced the secretion of inflammatory cytokines compared with the model group ( P < 0.01). LIG also upregulated miR?181b expression and downregulated its target gene, NF?κB. Inhibition of miR?181b mostly reduced the effect of LIG on inflammation in HUVECs. Conclusions LIG reduces damage by ox?LDL in HUVECs, partly through inhibiting expression of NF?κB via miR?181b to reduce the inflammatory reaction induced by ox?LDL.

Abstract:Objective To evaluate the effect of sevoflurane on the proliferation of neural stem cells (NSCs) in the ventricular/ subventricular zone of neonatal rats. Methods Forty?eight 7?day?old Sprague?Dawley rats, weighing 15 -20 g, were divided into three groups (n =16 each) using a random number table: control group (Con), 1% sevoflurane group (S1), and 3% sevoflurane group (S2). Rats in the control group inhaled air for 4 h. Rats in S1 and S2 groups inhaled 1% and 3% sevoflurane, respectively, for 4 h. Cell proliferation was detected by Nestin and 5?bromodeoxyuridine staining. Protein expression of Nestin, Cyclin D1, BMP2, and Smad4 in ventricular/ subventricular tissues was analyzed by Western blotting. Results Compared with the control group, the proliferation rate of NSCs was decreased significantly. Expression levels of Nestin and Cyclin D1 were downregulated, but expression levels of BMP2 and Smad4 were significantly upregulated after sevoflurane treatment ( P < 0.05). Conclusions Sevoflurane inhibits proliferation of NSCs in the ventricular/ subventricular zone of neonatal rats and the mechanism may be related to regulation of Cyclin D1, BMP2, and Smad4 expression.

Abstract:Objective To investigate the effect of focal cerebral ischemia/ reperfusion injury on TMEM166 and neuron autophagy. Methods In total, fifty C57/ BL6J mice were used to establish the focal cerebral ischemia model through middle cerebral artery occlusion (MCAO) by endovascular suture and were randomly divided into four groups according to different reperfusion time points: 6, 12, 24 and 48 h. Changes in the expression of TMEM166 and the number of LC3?II positive cells in cerebral cortex and basal ganglia of the ischemic hemisphere of the mice were measured by immunochemistry. Protein levels of TMEM166 and LC3?II were detected by Western blotting. Results TMEM166 was induced in mice after cerebral ischemia/ reperfusion injury, peaking at 24 h after injury in the ischemic hemisphere. LC3?II began to increase at 6 h following MCAO, increasing over time to reach a highest level at 24 h after ischemic insult. The changing pattern of TMEM166 was consistent with changes in LC3?II positive cell numbers. No changes of TMEM166 and LC3?II were found in the contralateral hemisphere cerebral cortex and basal ganglia. Conclusions TMEM166 is induced after MCAO, and can lead to neuron autophagy by activating LC3?II.

Abstract:Objective To investigate the effect of in utero intraperitoneal injection of human tumor HepG2 cells on the growth and development of fetal mice. Methods Pregnant mice were randomly divided into five groups and various densities of HepG2 cell suspensions were injected into the fetus by intrauterine injection at embryonic day 14. Sham operation and control groups were included. Pregnancies and the body weight of offspring were recorded, and developmental indicators were evaluated. Adult mice were selected and dissected to observe malformations and pathological changes in major organs including the liver, lungs, and kidneys. Results There was no significant difference in the growth or development of offspring in HepG2 cell injection and control groups. There were no lesions in major organs based on macroscopic and microstructural observations. Conclusions In utero intraperitoneal injection of human tumor HepG2 cells has no effect on the development of fetal mice.

Abstract:Objective To establish a stable, effective, and reproducible acute promyelocytic leukemia model in severe combined immunodeficient ( SCID) beige mice using acute myelocyte HL60 and adriamycin ( ADR)?resistant HL60/ ADR cell lines. Methods Female SCID beige mice (4 – 5 weeks old) were divided randomly into one control group and four model groups with three mice in each group. After X?ray irradiation, SCID mice received 1 × 107 (M1 group) or 5 ×106 (M2 group) HL60 cells, or 1 × 107 (M3 group) or 5 × 106 (M4 group)HL60/ ADR cells via tail vein injection. The white blood cell (WBC) count and positive rate of promyelocytes in peripheral blood were dynamically monitored by detecting cells expressing CD33 using flow cytometry. Morphological examination and histopathological assays were employed to confirm promyelocyte infiltration into organs (liver, spleen, and kidneys). Results The four model groups displayed abnormal behaviors of tremors, retardation, and piloerection. The two high dose model groups experienced significant weight loss compared with the two low dose model groups ( P < 0.05). Moreover, the four model groups had significantly lower body weights than the control group ( P <0.01). At day 32, survival rates of M1 and M3 groups were 33% and 67%, respectively. All mice in M2 and M4 groups survived. The WBC count in peripheral blood declined after X?ray irradiation. At day 28 after inoculation, the peripheral blood WBC counts of the four model groups were significantly higher than that in the control group ( P <0.01). At day 28, the leukemic cell percentage of the M3 group was the highest compared with the other three model groups ( P <0.05). The rates of CD33?positive cells in flow cytometric analysis of the four model groups were higher than that in the control group at day 28, but there were no significant differences between the four model groups. Morphological examination and HE staining of tissue biopsies demonstrated a large number of promyelocytes in the spleen and liver. Conclusions The human acute leukemia SCID beige mouse model was successfully established by tail vein injection of 5 × 106 or 1 × 107 HL60 or HL60/ ADR cells. This model mimics the biological characteristics of acute myeloid leukemia. The survival period of the high dose SCID beige mouse model was short with a median survival time of about 29 days.

Abstract:Objective To study effects of Ca, Mg, Cu, Zn, VD combined with protein on ovarian functions in female rats with coal?burning fluorosis. Methods A total of 120 clean?grade SD female rats with body weights of about 80 g were used. Rats were randomly divided into control, fluorosis, element, and element + protein groups with 30 rats in each group. Model groups were fed edible raw coal mixed with peat and different formulas of zhijin roasted corn feed. Rats were sacrificed at 60, 120, and 180 days, and bilateral ovaries were harvested. Expression of apoptosis proteins Bcl?2 and Bax in ovarian granulosa cells was observed by immunohistochemistry. HE staining was used to observe ovarian tissue morphology and changes in follicle and corpus luteum numbers. Results There was no statistical significance in the protein expression of Bcl?2 in ovarian granulosa cells. The expression of Bax in each group was compared with that in the fluorosis group, and a significant difference was found at 120 and 180 days ( P <0.05). There were significant differences at 180 days between the two intervention groups ( P <0.05). Compared within each group, Bax expression in the fluorosis group gradually increased and the difference was significant ( P <0.05). There was a significant difference at 60 and 180 days for various elements ( P < 0.05). Over a prolonged time in the fluoride group, ovarian granulosa cells showed progressive edema, atretic follicles, and corpus luteum degradation, and mature follicles were decreased significantly. Ovarian granular cells in the two intervention groups showed rare mild edema and normal numbers of follicles at all levels. Atresia of follicles and degeneration of the corpus luteum were decreased significantly. Compared with the fluorosis group, there were significant differences in atresia follicles between control and multiple element + protein groups at 60 days, and the control group and two intervention groups at 120 and 180 days ( P <0.05). Conclusions Ca, Mg, Cu, Zn, and VD antagonize the reproductive toxicity of coal?burning fluorosis and reduce protein expression of Bax and the number of atresia follicles. And the effects of intervention combined with protein can be better.

Abstract:Objective To determine the effect of reed rhizome polysaccharides (R?Poly) on hyperlipidemia in rats. Methods In total, 32 male SD rats were randomly divided into four groups: control group, model group, R?Poly group, and Lipitor group. Rats in the control group were fed a standard diet, while those in the other three groups received a high?fat diet. After eight weeks, rats in the R?poly group and Lipitor group were gavaged daily with 200 mg/ kg R?Poly or 2 mg/ kg atorvastatin for six weeks. The normal group and model group received an equal volume of physiological saline. At the end of the sixth week, all rats were terminated. Serum lipids were detected, and liver and aorta pathological examinations performed. Result Compared with the control group, serum triglyceride ( TG) levels were significantly decreased in the model group, while levels of total cholesterol (TC), low?density lipoprotein?cholesterol (LDL?C), and high?density lipoprotein?cholesterol (HDL?C) were significantly increased ( P < 0.01). Compared with the model group, levels of TC, LDL?C, and HDL?C were visibly decreased ( P < 0.01) in the R?Poly group and Lipitor group. Extensive fatty degeneration and inflammatory cells were found in model group rats, with the degree of hepatic steatosis and inflammation significantly alleviated in the R?Poly group and Lipitor group. Conclusions R?Poly can reduce the level of TC and LDL?C in rats with hyperlipidemia, and alleviate the degree of hepatic steatosis and inflammation. Accordingly, R?Poly has a good regulating effect.

Abstract:Objective To investigate the mechanisms underlying cognitive impairment induced by chronic aluminum exposure through observing the relationships between learning and memory functions and levels of ACh, β?EP, SOD, and MDA in the mouse cerebral cortex. Methods Mice were fed diets containing various doses of AlCl3. Their learning and memory abilities were measured by the Y maze test, and levels of ACh, β?EP, SOD, and MDA in the mouse cerebral cortex were measured at end of the experiment. Results The learning times and memory errors in groups administered moderate and high doses of aluminum were significantly increased compared with those in the control group. The levels of ACh, β?EP, and SOD in the cerebral cortex were decreased significantly, whereas the MDA level in the cerebral cortex was increased in groups administered moderate and high doses of aluminum compared with the control group. ACh, β?EP and SOD levels in the cerebral cortex were negatively correlated with learning times and memory errors in mice administered moderate and high doses of aluminum. MDA levels were positively correlated with learning times and memory errors in mice administered the high dose of aluminum. Conclusions Aluminum reduces the abilities of learning and memory in mice, possibly through decreasing the levels of ACh, β?EP, and SOD, and increasing the MDA level in the cerebral cortex, which may be the mechanisms underlying aluminum?induced cognitive impairment in mice.

Abstract:Objective To study the therapeutic effects of Alisma orientale extract (AOE) on nonalcoholic fatty liver disease in rats and systematically determine the underlying mechanism. Methods Collected samples of an oleic acid?palmitic acid mixture were applied to adipohepatic HepG2 cells after AOE treatment. ER stress marker proteins GRP78, CHOP, and XBP?1, as well as JNK1, p?JNK1, and STAT3 were detected by Western blotting. Serum SOD, AST, ALT, TG, HDL, and LDL levels were measured after establishment of high fat diet?induced nonalcoholic fatty liver disease and AOE treatment in SD rats as confirmed by oil red staining. CYP2E1 and CYP2A5 gene expression levels were determined in liver tissue by RT?PCR. Results After AOE treatment, JNK1, p?JNK1, GRP78, CHOP, and XBP?1 expression showed obvious dose?dependent downregulation, while STAT3 expression was increased. Serum AST, ALT, TG, and LDL levels were reduced dose?dependently, but SOD and HDL levels were increased. CYP2E1 and CYP2A5 gene expression levels were decreased obviously in liver tissue, especially in the high dose group. Conclusions AOE inhibited ER stress in adipohepatic HepG2 cells, and JNK signaling pathway and metabolic enzymes in vivo to treat nonalcoholic fatty liver disease.

Abstract:Objective To explore expression of GLUT?1 and HIF?1α in human colorectal carcinoma tissues and discuss its clinical significance. Methods Fifty specimens of colorectal carcinoma tissues ( cancer group) and 50specimens of normal adjacent tissues (normal group) were selected, and immunohistochemistry was used to detect the expression levels of GLUT?1 and HIF?1α in the two groups. Correlations between their expression levels and clinicopathological characteristics were analyzed, and the mechanism of carcinogenesis was investigated. Results Relative expression levels of GLUT?1 in cancer and normal groups were (2.285 ±0.459) and (1.086 ±0.148), respectively, and the difference was statistically significant (t =8.919, P <0.001). The relative expression levels of HIF?1α in cancer and normal groups were (2.203 ±0.401) and (1.055 ± 0.127), respectively, and the difference was statistically significant (t = 7.144, P < 0.001). GLUT?1 expression levels were positively correlated with expression levels of HIF?1α in colorectal carcinoma tissues (r = 0.958, P < 0.001). Expression of GLUT?1 and HIF?1α was correlated with the TNM stage, depth of tumor invasion, differentiation degree, lymph node metastasis, liver metastasis, and vessel invasion ( P <0.05). Conclusions The expression levels of GLUT?1 and HIF?1α are higher in colorectal carcinoma tissues, which serve as prognostic indictors for colorectal carcinoma.

Abstract:Objective Isolation, culture, and identification of dopaminergic neurons from newborn tree shrew to establish a cellular model and study abnormal dopaminergic neurons. Methods B27 and Glutamax were added to culture medium for in vitro culture of dopaminergic neurons from tree shrew. The cultured cells were identified by immunofluorescence. Results Cells isolated from newborn tree shrew were identified as dopaminergic neurons. The rate of tyrosine hydroxylase (TH)?positive neurons was (88.49 ± 2.4)%. Conclusions This method can be used to culture dopaminergic neurons from tree shrew in vitro.

Abstract:Objective To provide a simple, convenient, and safe anesthesia method for long?term operations in macaques. Methods Ten male Macaca mulatta macaques were anesthetized by a combination of intravenous anesthesia with propofol and isoflurane inhalation. The induction time, duration of anesthesia, and recovery time were recorded. After continuous monitoring of blood oxygen saturation, respiratory rate, and heart rate, the feasibility and effectiveness of the anesthetic scheme was evaluated by data analysis. Results All 10 monkeys survived the experimental anesthesia protocol. The anesthesia induction time was short and the onset was rapid. During anesthesia, vital signs were stable, and the sedation, analgesia, and muscle relaxation were satisfactory. Conclusions Combined anesthesia induced by propofol and isoflurane is a safe and effective method for long?term anesthesia of macaques.

Abstract:Acid calponin is an actin filament?associated regulatory protein expressed in smooth muscle and non?muscle cells. The roles of acid calponin in regulating dendritic spine remodeling, cytoskeletal rearrangement, smooth muscle contraction, skeletal muscle development, placentation processes, embryonic development, and signal transduction have been investigated extensively. These roles indicate that changes in acid calponin expression are related to the occurrence and development of drug?resistant epilepsy, colorectal cancer, mucosa?associated lymphoid tissue lymphoma, and ovarian cancer, which are significant for diagnosis and prognosis.

Abstract:Retinal bipolar cells are intermediate neurons in the signal pathway of the mammalian visual system. They receive signal inputs from photoreceptors (cone and rod cells) and transmit information to amacrine and ganglion cells after integrating information. Recent studies have shown that, through immunocytochemical staining, biophysics, pharmacology, membrane capacitance measurement, fluorescent biosensors, and two?photon microscopy, at the point where the visual path first diverges, a more comprehensive understanding of the entire category of neurons in the range of arrival can be obtained. Here, this article reviews the research progress in comparisons of the unique physiological structures and functions of different mammalian retinal bipolar cells and synaptic remodeling by immunocytochemical methods.

Abstract:Functional dyspepsia (FD) is a common clinical gastrointestinal functional disease. Establishment of an FD rat model will facilitate in?depth study of the disease. The primary method to induce the model employs single factors, but its limitations become obvious when the etiology of FD has been researched continually. However, compound factors have been gradually applied to induce the model, because they have the advantage of variously simulating the etiology.

Abstract:Marmosets, a kind of small non?human primates, are very suitable for research by gene modification because of their lower feeding cost and higher efficiency,shorter time of sexual maturation than macaques. Any model established by gene modification cannot be produced without assisted reproduction technology (ART) consisting of semen collection, superovulation, oocyte retrieval, in vitro maturation, in vitro fertilization, and embryo transplantation. At present, the research of these techniques is relatively comprehensive, but there are still details that need improvement. Recently, studies of gene?modified marmosets based on ART have gradually become a research focus with the development of gene editing technology. In 2009, the first report of transgenic marmosets was published by Japanese scientists, whereas related reports have not been published by Chinese scientists. This article reviews the recent progress of marmoset ART and their gene?modified models.

Abstract:Osteoarthritis is a major degenerative disease that affects human health. It causes a large number of physical disabilities and medical resource expenditure. However, the specific pathogenesis for this disease has not yet been clarified, and thus, it cannot be cured. The genes encoding sterol regulatory element binding proteins (SREBPs) have been studied in recent years. They play an important role in maintaining the homeostasis of lipid metabolism. Accumulating studies have confirmed that the genes are closely related to osteoarthritis. This article introduces the SREBP genes and discuss the relationship between SREBPs and osteoarthritis, and how SREBPs act in the mechanisms of metabolic disorders and osteoarthritis through related factors to provide a foundation for relevant research.

Abstract:The starry sky pattern is a distinctive histological feature in lymphoid tissue and lymphoma. The cytoplasm of these histiocytes typically contains cellular remnants. Here, this review discusses the historical, pathophysiological and pathophysiological research of the starry sky pattern, which may contribute to establishment of a lymphoma animal model, development of radiation and drug therapies, and determination of doses.

Abstract:Reactivation of cytomegalovirus ( CMV) can induce T cells to undergo immune senescence, a mechanism of aging. After CMV infection of immunocompetent hosts, it persists in bone marrow. CMV is stimulated and propagates into circulation via differentiation?dependent reactivation throughout life. This article compares and reviews the immune status of human and nonhuman primates after CMV infection, the immune escape mechanisms, and the relationship with immune senescence. Such information may provide a basis for establishing models of immune senescence and the development of drugs and vaccines.