Abstract:Objective To compare the characteristics of massive tibial bone defect (MTBD) models and induced membrane established by the internal fixation plate (IFP), intramedullary pin (IMP), and circular external fixator (CEF)based on Masquelet technology and selecting the optimal model method. Methods A total of 60 10-week-old SD male rats of SPF grade were randomly divided into three groups,namely, the IFP group, IMP group, and CEF group. A 4-mm bone defect model was generated at the middle of the right tibia, which was fixed with a custom-made six-hole stainless steel plate, a 1-mm-diameter Kirschner wire, and a custom-made circular external fixator, respectively. The time of modeling,the amount of bleeding, and the duration of limb swelling were recorded. X-ray examination was performed to observe the stability of bone cement and fixation devices. HE staining was conducted to observe the histomorphological characteristics.Results In terms of modeling time, bleeding volume, and model success rate, the IMP and CEF groups were significantly better than the IFP group, and significant differences were observed( P < 0. 05). The duration of limb swelling was the shortest in the CEF group, but no significant difference was found ( P > 0. 05). In addition, one case of screw loosening and three cases of bone cement loosening occurred in the IFP group, and 1 case of cement loosening occurred in the CEF group, whereas bone cement was well fixed in the IMP group. With regard to infection, there were three cases of plate exposure in the IFP group, two cases of purulent mass in the IMP group, and no infection in the CEF group. The thickness of induced membrane in the three groups was 460 -520 μm, showing no significant difference ( P >0. 05). Conclusions Taking the findings together, the three fixation models can successfully construct the MTBD model, but CEF is optimal for constructing the MTBD model by simulating the Masquelet technology.

Abstract:Objective To investigate the mechanism underlying catch-up growth diet-induced metabolic syndrome and brain aging symptoms in C57 mice. Methods Forty C57 mice were randomly divided into four groups; a normal control group (n =10), a calorie-restricted group (n = 10), a high-energy-diet group (n = 10) and a catch-up growth group (calorie-restricted/ high-energy-diet group, n = 10). These animals were continuously fed for 12 weeks. The body weight and blood glucose of the animals were recorded, biochemical indicators of metabolic syndrome were detected at the end of the experiment, HOMA-IR was calculated, the expression levels of senescence-associated protein P53 and phosphorylated P53 (ser15) were determined by Western blot, and the lipofuscin deposited in the hippocampus was observed by electron microscopy. Results Compared with the normal control group, the calorie-restricted group had lower body weight, blood glucose, and biochemical indicators, its levels of P53 and phosphorylated P53 protein expression were downregulated, and lipofuscin deposition was reduced. Meanwhile, compared with the normal control group, the metabolic syndrome indicators of the high-energy group and the catch-up growth group were significantly higher, the levels of P53 and phosphorylated P53 protein expression were increased, and more lipofuscin deposition was observed. Furthermore, the catch-up growth group showed more pronounced insulin resistance and brain aging symptoms. Conclusions A catch-up growth diet can induce metabolic syndrome in mice with brain aging. This study provides new research ideas for the construction of animal models with metabolic syndrome and neurodegenerative dysfunction caused by different dietary patterns.

Abstract:Objective To establish a high-fat-diet-induced heart failure model in Drosophila melanogaster.Methods Wild-type Drosophila w1118 was chosen as the experimental subject. The triglyceride level was examined in Drosophila fed with different concentrations of coconut oil (0%, 7. 5%, 15%, 30%). In the follow-up experiment, a high-fat diet containing 30% coconut oil was used for the experimental group, in contrast to a non-fat diet (0% coconut oil) for a control group. Using Drosophila negative geotaxis assay, the physical activity was evaluated. The pulse was recorded for 30 s and analyzed. Adult heart physiological parameters, including heart period, systolic and diastolic diameters, as well as fractional shortening, were obtained. The RNA expression of metabolism-related genes was measured by real-time PCR. Results The triglyceride content in high-fat-diet-fed Drosophila increased gradually with the concentration of coconut oil ( P < 0. 05). High-fat-diet-fed Drosophila showed dramatically decreased physical activity ( P < 0. 001), as determined using the negative geotaxis assay, and compromised adult physiological heart function,characterized by a decreased heart period and fractional shortening ( P < 0. 05). Metabolism-related genes, including fa2h, CG6277, CG3699, CG9914, and lip2, were downregulated in high-fat-diet-fed Drosophila compared with the levels in those with a non-fat diet. Conclusion The high-fat diet compromises physical activity and adult heart physiological function, revealing that a high-fat-diet-induced Drosophila heart failure model is successfully established here.

Abstract:Objective To investigate the effects of inflammatory factors, lipid metabolism, and glucose metabolism on weight in the northern pig-tailed macaques in acute stage of SIVmac239 infection. Methods The levels of inflammatory factor (TNF-α, IL-6), lipid metabolism (cholesterol, triglyceride, high-density-lipoprotein cholesterol, and low-density-lipoprotein cholesterol), and glucose metabolism parameters (insulin, insulin resistance index) were analyzed and compared between the weight increase group and the weight decrease group in the northern pig-tailed macaque in acute stage of SIVmac239 infection. Pearson’s correlation analysis was used to investigate the correlation of plasma viral load and insulin resistance. Results There was no difference of plasma lipid level and inflammatory factors between the weight increase group and the weight decrease group in the acute stage of SIVmac239 infection. After SIV infection, the insulin levels decreased in the weight increase group. In the weight decrease group, insulin levels were significantly elevated and insulin resistance appeared. Pearson’s analysis showed that changes of insulin resistance index at the 5^th week and 11^th week after infection were positively correlated with plasma viral load. Conclusion In the acute stage of SIVmac239 infection,abnormal glucose metabolism is the main cause of body weight change in northern pig-tailed macaques.

Abstract:Objective To observe the effect of lentiviral vector-mediated HCV receptor OCLN / CD 81 gene transfection on the biological characteristics and gene expression of tree shrew bone marrow mesenchymal stem cells (BM-MSCs) under in vitro culture conditions. Methods BM-MSCs were obtained by our previously described isolation method.We constructed a lentivirus vector containing the human OCLN / CD 81 gene and transfected it into BM-MSCs. The transfection efficiency was determined by fluorescence inverted microscopy and flow cytometry, and the proliferation of BMMSCs was determined using CCK8. Multipotent differentiation was induced by osteoblasts and adipocytes. The gene and protein expression of OCLN / CD 81 was detected by RT-PCR and WB, respectively. Results The transfection efficiency levels of LV- CD 81 and LV- OCLN were 99. 4% and 22. 6%, respectively, on MOI 100. The cell proliferation trends were similar to those of the untransfected BM-MSCs. The BM-MSCs were still undergoing osteogenic and adipogenic differentiation, but this declined. The pluripotency marker of NANOG mRNA was increased ( P < 0. 05), while the level of LIN28A mRNA was decreased ( P < 0. 05). BM-MSCs could express the transferred gene efficiently after transfection.Conclusions The constructed lentivirus vector containing the human OCLN / CD 81 gene can be successfully transfected into tree shrew BM-MSCs and the target gene is highly expressed. The multipotent differentiation potential of tree shrew BM-MSCs is clearly affected after transfection.

Abstract:Objective In a rabbit model of acute myocardial infarction, blood perfusion of the infarction segment after acute myocardial infarction was subjected to myocardial acoustic contrast visual evaluation. Methods Twenty cleangrade Japanese rabbits were randomly assigned to an acute myocardial infarction model group (group A) and a sham operation control group (group S). Preoperative and 10- and 45-min postoperative electrocardiograms (ECGs) were collected, and routine ultrasound and ultrasonic imaging were performed at 20 min, 2 h, and 6 h before and after the operation. Blood perfusion in the molding segment after acute myocardial infarction was comprehensively evaluated. At the end of the experiment, the myocardial tissue of the molding section was excised for pathological examination. Results Conventional echocardiography showed significant changes in the magnitude of the motion of the modular segment at different time points after ischemia, and significant changes in cardiac histology, indicating the successful establishment of the acute myocardial infarction model in Japanese rabbits. At the same time, visual evaluation of myocardial contrast sonography showed that the myocardial blood volume and perfusion speed of group A were significantly lower than those of the noninfarcted segments and the corresponding segments of group S, which was consistent with the result of routine echocardiography and pathological histology. Conclusion Visual evaluation of myocardial sonography can accurately determine the status of blood perfusion in the myocardial segment after acute myocardial infarction, which is an effective diagnostic method.

Abstract:Objective To develop a scan sequence to reduce respiratory motion and heartbeat artifacts in 256-slice spiral CT in living rat chest CT scans. Methods Using a Philips iCT, chest CT imaging was performed on 10 rats.For each rat, both A and B scanning methods were used: A mode for a conventional chest CT scan sequence, non-ECG gating spiral scanning mode, and B mode for non-ECG gating axis scanning mode. Both scanning method were performed under 120 kV and the scan length was 8 cm. Reconstruction of transverse and coronal lung windows and soft-tissue windows was performed on the obtained CT data. The images were observed and measured. Differences in mean CT values, image noise, signal-to-noise ratio (SNR), and subjective image quality scores of the two groups of images were analyzed. In addition, the consistency of the subjective image quality scores was tested among different observers. Results A total of 10 sets of rat chest CT data were analyzed. The DLP values of the scanning sequences of A and B were 144. 7 mGy/ cm and 72. 2 mGy/ cm. There was no significant difference between the CT values of lung tissue and of liver tissue between the two groups (( t = - 0. 205 and 0. 545; P > 0. 05). There was no statistically significant difference in noise between the two groups (( t = - 0. 865, P > 0. 05). There was also no statistically significant difference in lung tissue SNR between the two groups (( t =0. 903, P > 0. 05). However, liver SNR was significantly higher in group B than in group A (( t = - 2. 885, P <0. 05). The correlation coefficient between the scores of two physicians was 0. 763, and the consistency of the diagnostic result was good. The subjective quality scores in groups A and B were 2. 5 ± 0. 53 points and 4. 3 ± 0. 67 points, which were significantly different (χ² = 14. 76, P < 0. 05). Conclusions The non-ECG gating axial scan mode can obtain approximately the same CT value, noise, and SNR as conventional chest CT scan (non-ECG gating spiral scan mode), but the use of non-ECG gating axial scan mode can effectively reduce respiratory and heartbeat artifacts in living rat chest CT scans, reduce radiation dose, improve image quality, and improve observers’ diagnostic confidence.

Abstract:Objective To observe the dynamic blood glucose levels caused by diet-induced weight gain or loss.Methods Eight-week-old male C57BL/6 mice were fed a high-calorie diet (60% fat) for 8 weeks, and then divided into an intervention group (IT) and an obesity group (OB). Mice in the OB group were fed a high-calorie diet throughout the study.A normal control group (CON) was also established and given normal chow. Limiting caloric intake reduced the weight of the IT group to that of the CON level ( P > 0. 05), after that resuming high-calorie feeding until the body weight rise back to the OB level ( P > 0. 05). Intraperitoneal glucose tolerance test (IPGTT) was performed to evaluate blood glucose metabolism,insulin tolerance test (ITT) for insulin sensitivity, and HE staining of the pancreas for islet morphological study. Results When IT group mice were again exposed to a high-calorie diet, body weight growth and fasting blood glucose rate were 2. 27times and 5. 13 times, higher than those before caloric restriction. At the end of the intervention, larger area under the blood glucose curve of IPGTT ( P < 0. 05), decreased insulin sensitivity ( P < 0. 05), larger body fat depots ( P < 0. 05), and smaller islet area were detected in comparison with those in the OB group. Conclusion Body weight fluctuations lead to an increased intraperitoneal fat mass, decreased insulin sensitivity, and impaired glucose tolerance.

Abstract:Objective To investigate the role of RIP3 (receptor-interacting protein 3), which is a key molecule in regulating programmed cell necrosis, in H1N1 influenza virus infection. Methods RIP3 knockout (RIP3 ^{- / -} ) and wild-type (WT) mice were infected by influenza virus H1N1 PR8 at a dose of 5. 25 × 10³ TCID₅₀ (50% tissue culture infective dose). Changes of clinical signs, survival, and body weight of those mice were monitored daily for 14 consecutive days. Six mice from each group were sacrificed at 3 and 7 days post-infection (d. p. i. ), from which whole lungs were harvested. Some of the lobes were fixed in 4% paraformaldehyde for histopathological assessment and the rest were split and stored at -80℃ for further analysis. Real-time quantitative PCR and cytometric bead array (CBA) were performed to detect viral loads in lungs and inflammatory cytokines in supernatants of lung homogenates. Results Both groups showed severe symptoms after the infection of PR8. The RIP3 ^{- / -}mice with PR8 infection showed a high survival rate (50%)compared with the control group (12. 5%) ( P < 0. 05). The body weight loss of WT was greater than that of RIP3 ^{- / -}mice from 3 d. p. i. , but there was no significant difference between these two groups. No significant differences in viral loads and lung weight to body weight ratio were also observed between the two groups at 3 and 7 d. p. i. Pathological changes in RIP3 ^{- / -} mice were less severe than those in WT mice. CBA detection assay revealed that the levels of proinflammatory cytokines in the lungs of RIP3 ^{- / -} mice were lower than those in WT mice. Conclusion RIP3 plays a pathogenic role in mice infected with 5. 25 ×10³ TCID₅₀of influenza virus H1N1 PR8 by promoting inflammatory responses.

Abstract:Objective To investigate the effects of antioxidant N-acetylcysteine (NAC) on oxidative stress and FoxO1 activity of the liver in type 2 diabetic rats. Methods Twenty-four male SD rats were randomly divided into control(C), diabetes without treatment (D), and NAC-treated diabetic groups (D + NAC) (n =8). A model of type 2 diabetes was established by feeding on a high-fat diet and the intraperitoneal injection of low-dose streptozotocin (STZ). NAC was given at a dose of 1. 5 (kg·d), and groups C and D were given an equal volume of normal saline by gavage for 8 weeks.The levels of plasma triglyceride (TG), free fatty acid (FFA), glutamic oxaline aminotransferase (AST), and alanine transaminase (ALT) were measured by an automatic biochemical analyzer, and the levels of dismutase (SOD), catalase(CAT), glutathione peroxidase (GSH-Px), and adenosine 5′-triphosphate (ATP) in liver tissues and malondialdehyde(MDA) in plasma and liver tissues were detected by commercial kits. Caspase-3 expression and FoxO1 activity in liver were analyzed by Western blot. Results The plasma levels of TG, FFA, AST, ALT, and MDA, and hepatic MDA production, caspase-3 expression, and FoxO1 activity in D group were significantly increased as compared with those in C group, while the hepatic activities of SOD, CAT, and GSH-Px, as well as ATP levels were significantly decreased. All of these changes in diabetic rats were significantly attenuated by NAC treatment for 8 weeks. Conclusion Antioxidant NAC attenuates diabetes-related liver dysfunction, partially as a result of reducing oxidative stress, mitochondrial dysfunction,and FoxO1 activation.

Abstract:Objective To observe the effects of electroacupuncture combined with methylprednisolone sodium succinate and electroacupuncture or methylprednisolone sodium succinate alone for intervertebral disc extrusion. The effects on the limb movement function and Nissl bodies of rat models after 24 h were analyzed, and the incidence of side effects in each group was analyzed to provide a basis for clinical application. Methods A total of 112 SD rats were randomly divided into sham operation group (Sham), model group (Model), electroacupuncture before methylprednisolone sodium succinate group ( EA + MP), electroacupuncture group ( EA), and methylprednisolone sodium succinate group ( MP). The intervertebral disc extrusion model was prepared using a custom-made silicone gasket compression method. Each group was treated separately, and the BBB limb motor function score or Nissl staining of spinal cord tissue was observed at different time points to observe the changes of nerve cell Nissl bodies, as well as side effects and mortality. Results 1) BBB scores showed an upward trend in all groups after treatment, while on the third day and seventh day in the EA + MP group, the average BBB scores of hindlimbs of the compression side were significantly higher than those of the EA group and the MP group ( P <0. 05). Moreover, on the 14th day, each group was significantly higher than the Model group ( P < 0. 05) and was close to the Sham group ( P >0. 05). (2) Nissl staining showed that, compared with the levels in the model group,the damage of nerve cells in each treatment group was mild, and the number of Nissl bodies was high. The average BBB scores of EA + MP group was superior to the other two groups in Nissl bodies protection. (3) The incidence of side effects was 30% in the MP group, 4. 16% in the EA + MP group, and 0% in the EA group. The mortality rate in the MP group was 20%, while that in the EA + MP and EA groups was 0%. Conclusions The three treatment regimens increase the number of Nissl bodies to varying degrees, maintaine neuronal cell morphology, and improved limb motor function in rats with intervertebral disc extrusion for 24 h (more than the time window for methylprednisolone sodium succinate). Among the regimens, EA + MP has the best effect, significantly reduce the side effects and mortality rate of MP therapy, and produce a better curative effect than EA alone.

Abstract:Objective The goal here was to conduct a preliminary evaluation of the efficacy of a conjugate (NIRG) of heptamethine carbocyanine dye (IR-783) and the chemotherapeutic drug gemcitabine for tumor fluorescent imaging and targeted therapy. Methods First, the liver cancer cell line Hep3B was used to verify the specific absorption of NIRG, and nude mice as models of tumors from a subcutaneous xenograft of Hep3B cells were prepared to observe the effect of the NIRG fluorescence imaging in this CDX model. Three subcutaneous PDX models of liver cancer cases (C64003, C34566 and B66873) were prepared to study the effect of NIRG tumor-targeted fluorescence imaging and targeted therapy. Simultaneously, the expression of the hepatoma-specific antigen AFP was observed by immunohistochemistry in tumor tissues of these models. In addition, the expression of the antigens HIF1α and OATP associated with the specific uptake of NIRF dye was observed. Results The results showed that the binding sites of NIRG in liver cancer cell line Hep3B cells were mitochondria and lysosomes. NIRG-specific tumor sites were found in the subcutaneous CDX model of liver cancer, and the NIRF signal was enhanced with the growth of the tumor. NIRF signal and the luminescence signal had a good correlation (R² =0. 99716). The IHC results showed that AFP was highly expressed in the three PDX models, and HIF1α and OATP were also highly expressed. The fluorescence imaging results showed that NIRG had good tumor targeting and could inhibit tumor growth, showing a good therapeutic effect. Conclusions The conjugate NIRG has characteristics suitable for both fluorescence imaging and tumor targeting to deliver chemotherapeutic drugs. It is potentially an effective novel imaging and targeting agent for treatment of human liver cancer.

Abstract:Objective To determine the changes in the expression of mannose receptor and some inflammatory cytokines in the cornea of tree shrews infected by Fusarium solanum, and explore the mechanism of fungal infection keratitis. Methods A tree shrew cornea was infected by Fusarium solanum using a contact lens-assisted method, The animal model was evaluated by anterior segment photography, analysis of pathological sections, and in vivo confocal microscopy. qPCR was used to determine the expression of MR (mannose receptor), dectin-1, IL-1β, IL-6, and TNF-α in the tree shrew keratitis. The primary corneal cells of tree shrews were cultured and an infection experiment was carried out in vitro. Results The success rate of establishing the tree shrew fungal keratitis model was 84%. The positive expression of MR and dectin-1 at the early stage of infection caused peaks in the expression of IL-1β, IL-6, and TNF-α;the expression of these factors was positively correlated with the severity of fungal keratitis in the tree shrews. There was no significant change in corneal epithelial cell infection before and after infection ( P > 0. 05). Conclusions The model of fungal keratitis in tree shrew is successfully established, and its symptoms are similar to those of human fungal keratitis.Type C lectin receptors may be involved in regulating the immune response to fungal infection. The immune system may play an important role in the development and progression of keratitis.

Abstract:Objective To investigate the protective effect of Bifidobacterium (BIFI) on liver function in rats with chronic alcoholic liver injury (CALI) and to explore the mechanism involved. Methods SD rats were randomly divided into a CALI group, metadoxine (90 mg/ kg) group, BIFI low- (500 mg/ kg), medium- (1000 mg/ kg), and high-dose(2000 mg/ kg) groups, and SIRT1 inhibitor Tenovin-6 (25 mg/ kg) group. The CALI group and a blank control group were given equal volumes of normal saline. After 8 weeks, liver function of each group was analyzed. Levels of TG and TC in liver tissue and serum were determined. The pathological changes of liver tissue were observed by hematoxylin-eosin staining. The expression of SIRT1 and chREBP in liver tissue was analyzed by western blotting. Results Compared with the control group, the liver function of the rats in the CALI group decreased significantly, and the levels of ALT and AST in the blood increased significantly ( P < 0. 05). In addition, liver tissue underwent fatty pathological damage. The levels of TG and TC in liver tissue and serum were significantly increased ( P < 0. 05), the expression of SIRT1 protein was significantly decreased ( P < 0. 05), and the expression of chREBP protein was significantly increased ( P < 0. 05).Compared with the CALI group, the liver function of the rats in the low-, middle-, and high-dose BIFI groups was significantly enhanced. The levels of ALT and AST in the blood were also significantly decreased ( P < 0. 05), the pathological damage of liver tissue was significantly reduced, and the levels of TG and TC in liver tissue and serum were significantly decreased ( P < 0. 05), the expression of SIRT1 protein was significantly increased ( P < 0. 05), and the expression of chREBP protein was significantly decreased ( P < 0. 05). All of the above effects could be reversed by the SIRT1-specific inhibitor Tenovin-6. Conclusions BIFI may inhibit the accumulation of lipids by regulating SIRT1/ChREBP expression and protect rats from chronic alcoholic liver injury.

Abstract:Objective This study aimed to investigate method for the optimal breeding and identification of vitamin D receptor (VDR) knockout mice, which could provide animal models for further research on VDR function and related diseases. Methods Based on VDR knockout mice previously obtained in our lab, four different mating methods were used to generate offsprings, the phenotypes and genotypes of which were evaluated and the reproductive ability of VDR knockout homozygous (VDR^{- / -} ) and heterozygous (VDR^{+ / -} ) mice were compared. And, VDR expression was analyzed at the mRNA and protein levels. Results Compared with VDR^{+ / +} mice, VDR^{- / -} mice had a significantly different phenotype, mainly in terms of having less hair, small size, and decreased mobility. The genotype distributions of the offspring mice produced by the four breeding method were basically in accordance with Mendel’ s law of inheritance.VDR^{- / -} cross-over had poor reproductive ability and could not be directly used to expand VDR^{- / -} homozygous knockout mouse strains, while VDR^{+ / -} cross-over could successfully obtain VDR^{- / -} mice. It was also determined that VDR knockout traits could be stably inherited by offspring mice, so VDR^{+ / -} cross-over is the best way to expand the strains.Conclusions This experiment has successfully identified the genotypes of offspring mice and established VDR knockout mouse strains, laying the foundation for subsequent research on VDR function.

Abstract:Objective To investigate a method for systematically evaluating a precocious puberty rat model with phlegm-dampness syndrome. Methods To observe the symptoms of female SD rats in five groups, a precocious puberty with phlegm-dampness group, a precocious puberty without phlegm-dampness group, a blank control group, a parental model group, and a parental control group, and to conduct an open-field test to explore method for systematically evaluating animal model with the disease. Results There was no significant difference in body weight gain between the parental model group and the control group ( P >0. 05). Body weight in the 4-week-old female offsprings of the precocious puberty with phlegm-dampness group was significantly lower than that in the blank control group ( P < 0. 05). The weight of the precocious puberty with phlegm-dampness group was also decreased than the precocious puberty without phlegm-dampness group ( P < 0. 05). Compared with the parental control group, the female SD rats in the parental model group had a distinctly smaller total distance and speed of motion ( P < 0. 05). The motionless frequency and motionless time were clearly higher in the parental model group than in the parental control group ( P < 0. 05), but there was no significant difference ( P >0. 05) in the groups of young rats. Conclusions A high-fat diet failed to increase body weight in female SD rats; therefore, body weight index may not be a key indicator for systematic evaluation of animal models of phlegmdampness syndrome. The change of ethology in the rat model of phlegm-dampness syndrome is similar to the clinical manifestation of this syndrome. The open-field test can be used to further evaluate the animal model of phlegm-dampness syndrome. Difference in behavior was not found among female offsprings. The precocious puberty with phlegm-dampness syndrome remains to be futher investigated.

Abstract:Objective To investigate whether Huang Qi-San Qi mixture can be used to treat chronic kidney disease(CKD)rats by improving the structure of the colon mechanical barrier. Methods Thirty male Sprague-Dawley rats were randomly divided into five groups: sham group, model group (I/ R group), low dose of Huang Qi-San Qi mixture group (L-HSM group), high dose of Huang Qi-San Qi mixture group (H-HSM group), and NiaoDuqing group (NDQ group). The model of chronic kidney disease (CKD) was established by right nephrectomy and left kidney ischemia reperfusion. The blood levels of creatinine and urea nitrogen were measured at 2, 4, and 8 weeks. At the fourth week, the different groups were given intragastric administration as follows: The L-HSM and H-HSM groups were given 18 g/ (kg·d)and 36 g/ (kg·d) of Huang Qi-San Qi mixture, respectively. The NDQ group was given 1. 56g/ (kg· d) of NiaoDuqing granules, while the sham and I/ R groups were given the same volume of distilled water. The rats were killed at the 8th week and their kidney and colon tissues were excised. Then, the renal function (serum creatinine and urea nitrogen) was determined and the expression levels of renal fibrosis index proteins (TGF-β1 and α-SMA), inflammatory factors IFN-γ and IL-6, and colonic epithelium tight junction proteins ZO-1, occludin, and claudin-1 were determined. Results Compared with the model group,the high dose of Huang Qi-San Qi mixture effectively restored the damaged renal function,decreased the expression of TGF-β1 and α-SMA proteins, and alleviated renal fibrosis. The high expression of IFN-γ and IL-6 in colonic tissue of the model group was effectively reduced by the low and high dose of Huang Qi-San Qi mixture, and the results of immunohistochemistry and western blot analysis of colonic epithelium tight junction protein showed that,compared with the control group, the expression of IFN-γ and IL-6 in colonic epithelium was higher than that in the sham group. The expression of tight junction proteins ZO-1 and occludin in the model group was significantly decreased, while Huang Qi-San Qi mixture restored the structure of the colon mechanical barrier to some extent. Conclusion Huang Qi-San Qi mixture can prevent and treat CKD rats by improving the structure of the colon mechanical barrier and reducing the expression of inflammatory factors.

Abstract:Objective To analyze the structure and composition of the intestinal flora of the wild tree shrew using the Illumina MiSeq sequencing platform. Methods The feces of three wild tree shrews were collected and investigated for diversity using the Illumina MiSeq sequencing platform. Results In total, 181,657 sequences and 624 OTU were collected after sequencing. According to the Shannon-Wiener curve, the sequencing data were reliable for all bacteria of the samples. The bacteria in the tree shrew feces were from 9 phyla, 17 classes, 31 orders, 56 families, 124 genera, and 172 species. Among them, 1) Firmicutes and Bacteroidetes had the highest abundance, 68. 38% and 20. 52%, respectively.2) The dominant classes were Bacilli and Bacteroidia, at 54. 81% and 20. 52%. 3) The abundance of Lactobacillales and Bacteroidales was higher, at 50. 01% and 20. 52%. 4) Dominant families were Streptococcaceae and Prevotellaceae, at 40. 52% and 12. 13%. 5) Lactobacillus and Streptococcus were dominant (20. 03%, 19. 62%). 6) Beneficial bacteria,such as Lactobacillus and Lactococcus, were relatively common, while Bifidobacterium was relatively rare. 7) 16S functional prediction showed an abundance of functional genes, such as those involved in amino acid transport and metabolism,carbohydrate transport, and metabolism. Conclusions The composition of tree shrew fecal microbiota shows a rich diversity. Many bacteria that are relatively abundant remain unidentified, so further study is warranted.

Abstract:Objective To investigate the feasibility of PLCζ as an activator and its effect on fertilization and embryo development for oocytes after mouse round spermatid injection. Methods The recombinant plasmid pCRII-TOPOPLCζ was constructed in vitro and its expression was induced. The protein was identified by mass spectrometry and the antigenicity was detected by Western blot. Recombinant PLCζ protein was used as an activator of mouse ROSI oocytes, and fertilization and embryo development were recorded and analyzed statistically. Results The recombinant plasmid pCRIITOPO-PLCζ was successfully constructed, and the recombinant PLCζ protein was identified by mass spectrometry and had good antigenicity. The fertilization rate, two- to four-cell rate, eight-cell rate, and cystic rate associated with the recombinant PLCζ protein did not differ significantly from those in the blank group ( P > 0. 05). Conclusions The recombinant PLCζ protein has no significant effect on oocyte fertilization and embryo development in this study. The feasibility of being an oocyte activator after mouse ROSI is questionable.

Abstract:Objective To study the effects of intervention on physiological and biochemical mechanism of psychodependent drug(heroin)addiction. Methods The rat models of heroin addiction were established by treated with heroin 135 mg/ kg to Wistar rats for 14 days, or treated with reserpine 2. 5 mg/ kg and then heroin, and the intervention effect of reserpine on heroin addiction was assessed. Levels of noradrenaline (NA), adenosine cyclophosphate (cAMP), guanosine cyclophosphate (cGMP) and hydroxybutyric acid (GABA), dopamine (DA) in blood and brain tissue of rats were determined by ELISA. Results The levels of cAMP in the heroin group and reserpine group were increased by 18. 87%and 35. 27%, and the cGMP levels were decreased by 10. 15% and 17. 69%, respectively. NA level in the heroin group was increased by 34. 05% than the control group and decreased by 45. 34% than the reserpine group. The levels of GABA in the frontal cortex (PFC), nucleus accumbens (NAc), thalamus (Thalamus) and amygdala (Amygdala) of the heroin group were reduced by 36. 55%, 40. 35%, 42. 34% and 45. 76%, respectively, compared with the control group ( P <0. 01), while the levels GABA in the four tissues of the reserpine group were increased by 31. 06%, 25. 91%, 49. 32 and17. 97%, respectively ( P < 0. 05, P < 0. 01). The levels of DA in the four tissues of the heroin group and reserpine group were increased by 82. 83%, 68. 19%, 87. 26%, 82. 46% ( P < 0. 01) and 8. 79%, 19. 06%, 21. 65%, 19. 49% ( P <0. 05), respectively, compared with the control group. After heroin and reserpine were given, there were significant differences in the frequency of EEG and ECG in the rats. Conclusions Noradrenaline and dopamine are the key factors to induce heroin addiction in vivo. Reserpine depletes noradrenaline and interferes with the biochemical process of heroin addiction.

Abstract:The patient-derived xenograft (PDX) model established by transplanting the patient’s tumor tissue into immunodeficient mice well preserves the biological characteristics of the primary tumor. However, the stroma component of the tumor tissue from patients is gradually replaced by the mouse-derived matrix over time in mice. The conditions in mice lacking an immune system will also change the microenvironment of the tumor, and the heterogeneity of the primary tumor will gradually be lost, thus limiting the application of the PDX model. The above problems can be resolved by a humanized patient-derived xenograft (Hu-PDX), which was established by transplanting human hematopoietic stem cells (HSCs) into immunodeficient mice, in which exposure to low-dose whole-body irradiation causes damage to bone marrow cells, or other method to clean up the bone marrow, forming humanized immune system mice (HISM). Then, the tumor tissues from patients can be transplanted into the HISM. HSCs in the mice can be induced to produce human immune cells. That makes the microenvironment of the tumor grown more similar to that in humans. This model well maintain the molecular heterogeneity of the primary tumor, and provides a good animal model for individualized tumor treatment.

Abstract:Transcranial ultrasound-assisted thrombolysis, also called sonothrombolysis (STL), is a method of treatment using ultrasound to promote clot dissolution and accelerate recanalization of occluded vessels in the brain with ischemic stroke. It has been demonstrated that STL can enhance the rate of thrombolysis without increasing complications such as symptomatic intracranial hemorrhage. Sonothrombolysis has been used in clinical trials for more than a decade, but has yet to be widely accepted. One of the main reasons is the lack of systematic and rigorous assessment of the safety and effectiveness of this approach. In this review, we summarize STL studies performed in animal models of ischemic stroke reported during the last 10 years and highlight some current issues and potential solutions.