Abstract:Objective　To obtain the full-length coding sequence of β-amyloid precursor protein-cleaving enzyme(BACE1) and analyze its molecular characteristics. Methods　Total RNAs from the brains and other organs of tree shrewswere used. The full-length coding sequence of BACE1 was obtained by splicing the sequence obtained via rapid amplificationof cDNA ends (RACE) with the intermediate sequence. Sequence and molecular characteristic analyses were performedusing DNAMAN, MEGA 10. 0. 5, PyMOL and other bioinformatics softwares. Results　The BACE1 nucleic acid sequenceof tree shrews was highly similar to that of humans. The overall structures of the two protein molecules were highly similarand had the same extracellular, transmembrane and intracellular domains. Both the tree shrew and the human BACE1proteins had a highly conserved ACDL sorting endocytic motif of DXXLL in the intramembranous domain. However, the treeshrew BACE1 protein had one potential acetylation site compared with that of humans and differed in its folding structure.Conclusions　The complete coding sequence of the BACE1 gene is obtained in our study, and the protein structure of treeshrews is similar to that of humans, suggesting that tree shrews may be an ideal model for studying pathological changes of Alzheimer’s disease or potential drug research.

Abstract:Objective 　To investigate the distribution of intestinal microflora in tree shrews with high-gradesarcomas and healthy tree shrews and compare their intestinal microbial structure and diversity, and to explore therelationship of intestinal microflora between high-grade sarcoma and healthy one in tree shrews. Methods　Feces from threetree shrews with malignant high-level sarcomas were selected as the CA group, and feces from 6 healthy tree shrews wereused as the control group (NO). Bacterial diversity was identified using primers in the 16S V3+4 region. The intestinalmicrobial structure, abundance and diversity were obtained for both groups by cutting and filtering reads, clusteringoperational taxonomic units (OTUs), and analyzing the species annotation and abundance. Results 　The intestinalmicrobial structure of the tree shrews in the CA and NO groups were compared, and the Chao1 diversity, ACE and Shannonindices did not show significant differences ( P >0. 05). The NO group contained 640 OTUs, and 494 OTUs were found inthe CA group. At the phylum level, the abundance of Spirochaetes in the CA group was 0. 03%, which was significantlylower than that in the NO group (0. 12%; P <0. 05). The abundance of Firmicutes in the CA group was 77. 79%, whichwas higher than that in the NO group (47. 65%), but the difference was not statistically significant. The Bacteroidetesabundance in the CA group was 0. 75% lower than that in the NO group (8. 31%), but this difference was also notstatistically significant. Lactobacillus was a statistically significant biomarker in the CA group. Conclusions　The structuralabundance and diversity of the intestinal microflora in tree shrews with high-grade sarcomas are lower than those in healthy tree shrews.

Abstract:Objective To establish a method for genetic detection of pigeon microsatellite DNA in a closed-groupexperiment. Methods Sixty-one pigeon microsatellite loci were selected and designed through literature search and SSR(simple sequence repeats) Hunter software screening. PCR amplification and condition optimization were performed usingpigeon genome samples. Twenty microsatellite loci with abundant alleles and distinct amplification bands were obtainedusing agarose gel electrophoresis. Through comparative analysis of STR (short tandem repeat) scanning, 16 loci suitable fortesting the genetic quality of experimental pigeons were selected and applied to two closed-group pigeon populations.Results Sixteen microsatellite DNA loci were selected to establish the genetic detection method of a closed group ofpigeons. Conclusions A genetic detection method based on microsatellite DNA is established for a closed group of pigeons. This method lays a foundation for genetic quality control in a closed-group pigeon population.

Abstract:Objective To establish a genetic monitoring method for laboratory chickens. Methods Seventy-twomicrosatellite loci were chosen from the literature. PCR and short tandem repeat (STR) scanning techniques were used toidentify the loci with good amplification effects and stability. The loci suitable for genetic monitoring of closed-colony andhaplotype laboratory chickens were preliminarily established. This genetic monitoring method was applied in three closedlaboratory chicken colonies and three haplotype laboratory chickens. Results We preliminarily determined 28 microsatelliteloci for genetic monitoring of closed-colony laboratory chickens and 14 microsatellite loci for genetic testing of haplotypelaboratory chickens. Conclusions A genetic monitoring method for laboratory chickens is preliminary established.

Abstract:Objective Traditional serological method for detecting the ABO blood group in humans are unsuitablefor macaques. Detecting the ABO blood group in macaques is important for transplantation and research. This study aimed toestablish a rapid and reliable nucleic acid-based method for identifying the ABO blood group in Macaca fascicularis andMacaca mulatta. Methods A new molecular method for ABO blood group detection was established, using competitiveallele-specific PCR (KASP) to identify the Macaca fascicularis and Macaca mulatta’ blood group by identifying the basesat 2 single nucleotide polymorphism (SNP) loci that are closely related to the blood-type functional gene. The bloodsamples identified via the KASP method were identified by Sanger sequencing by designing PCR primers around the twoSNP loci. Results Fifteen whole-blood nucleic acid samples from Macaca fascicularis and Macaca mulatta were determinedby TaqMan PCR and compared. The newly established KASP typing result were consistent with the original blood typeresult. To ensure the accuracy of the KASP method identification result, we used the newly established PCR method verifiedby Sanger sequencing, showing that the KASP method was consistent with the sequencing results. To verify the repeatabilityof the new KASP method, six blood samples with known blood groups were selected, and five replicates were taken perblood sample. The results showed that the method had excellent repeatability. In addition, 51 clinical samples were detectedusing the new KASP method, including 38 samples from cynomolgus monkeys and 13 from rhesus monkeys. The blood typeresults for two sites from the same sample were consistent, of which, 9 were type A (17. 6%), 19 were type B (37. 3%),23 were type AB (45. 1%), and none was type O. From the clinical samples with types A, B, and AB, five of each wereselected for PCR amplification and sequencing analysis, which confirmed that the sequences at the two SNP loci wereconsistent with those of the KASP typing. Conclusions The new detection method using KASP is highly accurate and canbe used to rapidly detect the ABO blood types in macaques. Compared with traditional serology and sequencing method, thismethod is simple, rapid, economical and reliable, and the results are easily interpreted. Furthermore, the samplerequirements are lower than those of traditional serological method, and fresh or long-stored whole blood, saliva or nucleic acid can be used.

Abstract:Objective The research of traditional Chinese medicine has gradually revealed a mechanism fortreating disease by regulating the microflora. The aim of this study is to establish an asthmatic mouse model with airwaymicrobial dysbiosis to provide a suitable model for studying the effects of Chinese medicine on respiratory flora and asthma.Methods Forty BALB/ c mice were randomly assigned into the control, house dust mite (HDM) asthma model, asthmamodel with intranasal administration of cefoperazone sulbactam sodium (CFP; i. n. CFP+HDM) and asthma model withintraperitoneal injection of CFP (i.p. CFP+HDM) groups. The i.n. CFP+HDM group and the i.p. CFP+HDM group wereestablished to observe the effect of antibiotics on HDM-induced asthma in mice. After 24 d of model establishment,eosinophils (EOS) in the peripheral blood ( PB) of the mice were counted. Serum IgE, TH2 cytokines in thebronchoalveolar lavage fluid (BALF) and in the lung homogenates were analyzed by ELISA. Pulmonary pathologicalchanges were observed using hematoxylin and eosin, periodic acid-Schiff and Masson staining. Results In contrast withthe control mice, EOS in the PB, serum IgE, TH2 cytokines in the BALF and lung homogenates in the HDM group weresignificantly increased ( P <0. 05), while these changes in the i. n. CFP+HDM and i. p. CFP+HDM groups were moresignificantly pronounced than those in the HDM group ( P <0. 05). Similarly, the pathological observation showed moresevere asthmatic changes in the i.n. CFP+HDM mice, but the pathology of the i.p. CFP+HDM group did not significantlydiffer from that of the model group. Conclusions Intranasally administered high-dose antibiotics, cefoperazone sulbactamsodium, aggravated HDM-induced asthma in mice, suggesting that administering antibiotics via the airway may disturb theairway microbiome and aggravate asthma occurrence. This model can be used to study Chinese medicine as a method of regulating airway microbiomes of asthma in germ-free mice.

Abstract:Objective To investigate the regulatory role of the clinical hepatinica, polyene phosphatidylcholine(PPC), on LPS-induced macrophage inflammatory response and whether phosphate and tension homology deleted onchromosome ten (PTEN) is involved in the process. The study also preliminarily determined the anti-inflammatory effect ofPPC and its underlying mechanisms. Methods Raw 264. 7 cells were inoculated on cell culture plates and divided into sixgroups, which were labeled and induced as follows: PPC (20 μg/ mL), LPS (100 ng/ mL), LPS (100 ng/ mL) + PPC(20 μg/ mL), PPC (20 μg/ mL) + SF1670 (150 ng/ mL), PPC (20 μg/ mL) + SF1670 (150 ng/ mL) + LPS (100 ng/mL) or an equal volume of phosphate-buffered saline (PBS) groups. The cells and supernatants were collected after 24 h.Enzyme-linked immunosorbent assay was used to detect the levels of IL-6, IL-10 and TNF-α. RT-PCR was applied to detectthe mRNA expression of IL-6. Western blot was performed to confirm the PTEN protein expression. Results TNF-α andIL-10 levels in the supernatants did not significantly differ between the PBS and PPC groups. Compared with the LPSgroup, the TNF-α and IL-6 levels in culture supernatants of the PPC+LPS group decreased significantly ( P <0. 001),while the IL-10 level increased significantly ( P <0. 001). PTEN protein expression in the PPC+LPS group was significantlyhigher than that of the LPS group. Compared with the PPC group, the TNF-α levels and IL-6 mRNA expression in the PPC+SF1670 group were significantly increased ( P <0. 05). The PPC+LPS+SF1670 group showed elevated TNF-α and IL-6production ( P <0. 001) but lower IL-10 production ( P <0. 001) than that of the PPC+LPS group. Conclusions PPC inhibits LPS-induced macrophage inflammatory response by upregulating PTEN expression.

Abstract:Objective To establish an in vitro culture system of adult marmoset kidney cells, and create a meansfor detecting single guide (sg) RNA cutting activity based on the Cas9 editing system. Methods A growth curve wasinitially determined for adult marmoset kidney cells. The growth curve, transfection reagent and resistance concentrationwere determined. These were then used to co-transfect SIRT1 sgRNA and Cas9 into marmoset cells. Following transfection,the genome was extracted and sequenced. Results The transfection rate of ViaFect was greater than 50% ~ 70%. Theoptimum concentration of puromycin was 4 μg/ mL and that of blasticidin was 8 μg/ mL for cell resistance screening.Sequencing result showed that mutation peaks appeared near the PAM sequence of sgRNA, which proved that sgRNA wassuccessfully introduced mutations into the marmoset genome. Conclusions We successfully established an in vitro culture,transfection and resistance screening system for marmoset kidney cells, which can be used as a reference for gene editing site testing. CRISPR/ Cas9 is suitable for renal cell editing in marmosets.

Abstract:Objective To evaluate the antiglioma effect and safety of extracts of the Chinese medicine, Jinlongcapsules (JLC), as well as the crude polysaccharides and proteins prepared from these extracts, to confirm the antigliomaefficacy and safety of the prepared JLC extracts. Methods One hundred 3-4-week old male BALB/ c-nu nude mice weredivided into the following groups: blank control (without tumor, solvent) negative control (solvent), positive control(temozolomide, 32 mg/ kg), low-dose JLC extract (750 mg/ kg), medium-dose JLC extract (1200 mg/ kg), high-doseJLC extract (1500 mg/ kg), crude polysaccharides (100 mg/ kg), and crude protein (100 mg/ kg) groups. An orthotopicsurgical technique was applied to implant a piece of human glioma U87-RFP tumor mass into the brains of nude mice toestablish an orthotopic implantation model of U87-RFP glioma. After building the model, a FluorVivo Model 100fluorescence imaging system was used for noninvasive real-time imaging of the brain tumors in the mice. The effects of JLCextracts on the growth of the orthotopic U87-RFP model were evaluated by comparing the tumor inhibition rates of thesegroups. Results After 25 days of intragastric administration of the drugs, the tumor inhibition rates of the low-dose andmedium-dose groups were both 34. 63%. and neither group’s weight differed significantly from that of the negative controlgroup. For the crude polysaccharide and crude protein groups, the tumor inhibition rates were 22. 26% and 29. 01%, andthe tumor weights were 0. 178±0. 048 g and 0. 144±0. 047 g, respectively, not significantly different from those of thenegative control group. For the positive-control and high-dose groups, the tumor inhibition rates were 99. 57% and61. 90%, and the tumor weights were 0. 001±0. 001 g and 0. 088±0. 045 g, respectively, which differed significantly ( P <0. 01 and P <0. 05, respectively). Conclusions Our findings indicate that crude JLC polysaccharides and proteins have certain inhibiting effect on gliomas. At the doses used in this study, no obvious acute toxic effect is observed.

Abstract:Objective To assess the clinical significance of histone deacetylase 6 (HDAC6) expression in nonsmallcell lung cancer (NSCLC) and determine the relationship between HDAC6 and the epithelial-mesenchymal transition(EMT). Methods Tumor and noncancerous peritumoral lung tissues (controls) were collected from 52 patients withNSCLC and analyzed for HDAC6 and E-cadherin expression using immunohistochemistry. Correlation analyses betweenHDAC6, E-cadherin and clinicopathological parameters were performed using χ2 tests. In vitro studies were conducted intransforming growth factor (TGF)-β1-induced A549 cells to determine the effect of HDAC6 overexpression on molecularmarkers of EMT. Results Immunohistochemical analysis revealed significantly elevated HDAC6 expression in NSCLCtissues, which correlated with the differentiation stage and lymph node metastasis. Moreover, HDAC6 expression wasinversely correlated with that of E-cadherin, an EMT marker. Treatment with an HDAC6 inhibitor and signaling inhibitors ofRho-associated protein kinase ( ROCK) or extracellular signal-regulated kinases ( ERK) significantly reduced theproliferation, migration and invasion of TGF-β1-induced A549 cells. Furthermore, western blot analysis indicated that TGF-β1 induced EMT in A549 cells, which was manifested by upregulated E-cadherin and vimentin and downregulated α-smooth muscle actin (α-SMA). Signaling inhibitors against HDAC6, ROCK and ERK reversed the effects of TGF-β1treatment. Conclusions Our data suggest that HDAC6 plays an important regulatory role in NSCLC tumorigenesis andprogression and is closely related to tumor EMT and regulation of ROCK and ERK signaling. Targeting HDAC6 activity using HDAC6, ERK1/2 or ROCK inhibitors may be a potential therapeutic option for NSCLC.

Abstract:Objective To investigate the protective effect of puerarin (Pue) on acute liver failure (ALF)induced by d-galactosamine in mice and its mechanism. Methods Fifty Kunming mice were randomly divided into fivegroups. Two weeks before modeling, the Pue group, LY294002 group (LY group), and Pue + LY group were injected with300 mg/ kg Pue, 10 mg/ kg LY, or 300 mg/ kg Pue + 10 mg/ kg LY, respectively, into the caudal vein once daily. Thenormal control and model group rats were administered the same amount of sterile physiological saline. After the finaladministration and fasting for 24 h, the ALF model was established by intraperitoneal injection of galactose in all groupsexcept the control group which received the same amount of sterile normal saline. Serum levels of alanine aminotransferase(ALT), aspartate transferase (AST), and total bilirubin (TBil) were detected by a biochemical analyzer, and levels ofmalondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in liver tissues weredetected by kits. Hematoxylin and eosin staining, TUNEL staining, and western blotting were used to detect pathologicalchanges in the mouse liver tissue, apoptosis in liver cells, and the expression levels of P-Akt, P-glycogen synthase kinase(GSK)-3β, and cleaved caspase-3 protein, respectively. Results Compared with the control group, significant increaseswere detected in the number of apoptotic hepatocytes, serum ALT, AST, and TBil levels, the liver tissue MDA content,and cleaved caspase-3 protein expression levels in the model group ( P < 0. 05), while protein expression levels of SOD,GSH-Px, P-Akt, and P-GSK-3β were significantly decreased ( P < 0. 05). After treatment, all indices in the Pue groupwere significantly improved compared with the model group ( P < 0. 05), while there were no significant differencesbetween indices among the LY group, Pue+LY group, and model group ( P > 0. 05). Conclusions Puerarin may play aprotective role in the liver of mice with d-galactosaminoglycan-induced acute liver failure by activating the PI3K/ Akt signaling pathway, thereby reducing the degree of liver function damage.

Abstract:Objective To study the effects of Dendrobium nobile Lindl. alkaloids (DNLA) on insulin resistancein diabetic rats with non-alcoholic fatty liver disease (NAFLD). Methods Sprague-Dawley (SD) rats were fed a high-fat,high-sugar diet for 4 weeks, and intraperitoneally injected with 40 mg/ kg streptozotocin to establish a diabetic rat modelwith NAFLD. Rats were then randomly divided into the model group, DNLA 20 mg/ kg group, DNLA 40 mg/ kg group,DNLA 80 mg/ kg group, and metformin 100 mg/ kg group (n = 10 rats per group). Ten normal SD rats were used ascontrols. Four weeks after intragastric administration of the drugs, fasting blood glucose (FPG) and fasting insulin (FINS)levels were measured, the insulin resistance index (HOMA-IR) was calculated, histological changes of the liver andpancreas were observed using hematoxylin and eosin staining, and islet insulin levels were detected byimmunohistochemistry. Results FPG, FINS, and HOMA-IR were significantly increased in the model rats compared withcontrols ( P < 0. 05), and FPG, FINS, and HOMA-IR were significantly decreased in the DNLA 40 mg/ kg and DNLA 80mg/ kg groups compared with controls ( P < 0. 05). Compared with the model group, the degree of fatty degeneration of theliver and the number of islet cells were reduced in diabetic rats. The expression of insulin in the islets of diabetic rats withNAFLD was significantly decreased. Compared with the model group, DNLA 40 mg/ kg and DNLA 80 mg/ kg groups showeda reduced extent of fatty degeneration of the liver and more islet cells. Insulin expression increased by varying degrees in theislets of the 40 mg/ kg and 80 mg/ kg DNLA groups. Conclusions DNLA reduces blood glucose and liver steatosis in diabetic rats with NAFLD. The mechanism may be related to the improvement of insulin resistance.

Abstract:Objective To investigate the changes in cardiac electrophysiology and cardiac function in rats afterlimb amputation and their relationships with the endothelial nitric oxide synthase/ nitric oxide ( eNOS/ NO) pathway.Methods Seventy-two healthy 8-week old male Wistar rats were divided into the normal group (anesthesia only),amputation control, amputation 0. 25 h, amputation 0. 5 h, amputation 0. 75 h, and amputation 1. 5 h groups (n=12 ratsper group). The left hind limb trauma model was established by amputation surgery. Changes in heart rate, QT interval,and PR interval were detected by electrocardiogram; the left ventricular ejection fraction (LVEF) and left ventricular shortaxis shortening rate (LVFS) were measured by echocardiography; the left ventricular systolic pressure (LVSP), thehighest rate of increase of left ventricular pressure (+dp/ dt max), and the highest rate of decrease of left ventricularpressure (-dp/ dt max) were measured by right carotid artery intubation; the level of malondialdehyde (MDA) in themyocardium was measured with thiobarbituric acid; the level of superoxide dismutase (SOD) was detected by pyrogallolcolorimetry; Green’s method was used to detect the nitric oxide (NO) levels; levels of myeloperoxidase (MPO), tumornecrosis factor-ɑ (TNF-α), and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay; hematoxylinand eosin staining was used to observe the histopathological changes in the myocardium; myocardial cell apoptosis wasobserved by TUNEL staining; and the expression of eNOS, Bcl-2, and Bcl-2-related X protein (Bax) in the myocardiumwas detected by western blotting. Results Compared with the normal group, the following changes were observed 0. 5 hand 0. 75 h after amputation: the heart rate increased, the QT interval decreased, LVSP, +dp/ dmax, -dp/ dmax, LVEF,and LVFS decreased, MPO, MDA, SOD, TNF-α, and IL-6 increased, and the myocardial cells were loosely arranged,necrotic, and accompanied with a large amount of inflammatory cell infiltration. Moreover, at these same time points, thefollowing significant changes were observed in a time-dependent manner ( P < 0. 05): an increased apoptotic index ofcardiomyocytes, decreased expression of Bcl-2 protein in the myocardium, increased expression of Bax protein, anddecreased NO level and eNOS protein expression. Compared with 0. 75 h after amputation group, the heart rate decreased at0. 5 h after amputation, the QT interval and PR interval increased, LVSP, +dp/ dmax, -dp/ dmax, LVEF, and LVFSincreased, MPO, MDA, SOD, TNF-α, and IL-6 decreased, and the degree of myocardial pathological injury wasalleviated. Moreover, significant differences were seen as following: a decreased apoptotic index of cardiomyocytes,increased Bcl-2 and eNOS protein expression, increased NO levels, and decreased Bax protein expression ( P <0. 05).Conclusions Amputation trauma causes ischemic electrocardiogram changes, cardiac function damage, excessiveoxidative stress, and inflammatory damages in rats. The underlying mechanism may be related to inhibition of the eNOS/ NO pathway.

Abstract:Objective To investigate the expression level of Fc receptor-like protein 3 (FCRL3) in hepatocellularcarcinoma and to determine its effect on the proliferation and migration of hepatocellular carcinoma HepG2 cells. MethodsThe expression of FCRL3 mRNA and protein was detected in hepatocellular carcinoma tissues, healthy liver tissues, andHepG2 cells by RT-PCR. The effects of FCRL3-specific small interfering (si)RNA on the proliferation and migration ofHepG2 cells were detected by an MTT assay and cell scratch test. Results FCRL3 mRNA and protein expression wassignificantly higher in hepatocellular carcinoma tissues than in healthy liver tissues ( P <0. 05). FCRL3 mRNA and proteinexpression levels were significantly lower following siRNA in HepG2 cells, and the proliferation rate and migration abilitywere significantly decreased ( P < 0. 05). Additionally, the expression of FCRL3 mRNA and protein in HepG2 cellsoverexpressing FCRL3 was significantly increased compared with controls, as well the proliferation rate and migrationability ( P <0. 05). Conclusions The expression of FCRL3 in hepatocellular carcinoma is up-regulated compared withhealthy liver tissue. FCRL3 inhibition reduces the proliferation and migration of hepatoma cells, suggesting that FCRL3 plays an important role in the progression of hepatocellular carcinoma.

Abstract:Objective To investigate the level of CD4⁺ T cells and related mechanisms in fetal heart tissue fromfemale rats with gestational diabetes mellitus ( GDM). Methods Eighty pregnant female Sprague-Dawley rats wererandomly divided into normal control (NC) group and GDM group induced by streptozotocin. CD4⁺ cell levels were detectedin the fetal heart tissues of different groups. Western blotting was performed to detect the interleukin (IL)-17, IL-4, T-cellimmunoglobulin mucin (Tim)-3, and programmed cell death protein (PD)-1 expressions, and real-time PCR was used todetect the expression of miR-233-3p. Results The expression of CD4⁺ CD25⁺ Treg cells in the GDM group (46. 41±10. 94%) was significantly higher than in the NC group (21. 63±10. 94%) ( P <0. 01). Expression levels of IL-17, Tim-3, and PD-1 were also significantly higher in the GDM group than in the NC group, while the expression of IL-4 wassignificantly lower ( P <0. 05 or P <0. 01). The relative expression of miR-233-3p in the GDM group (0. 682±0. 104) wassignificantly lower than in the NC group (0. 466±0. 117) ( P <0. 05). Conclusions Cardiac tissue abnormalities in GDMrats may be associated with the down-regulated expression of CD4⁺ T cells, and the underlying mechanism may be associated with the down-regulation of miR-233-3p expression in the fetal rat heart tissue.

Abstract:Objective To explore the effect of matrine combined with an Akt signaling pathway inhibitor on theproliferation and apoptosis in renal carcinoma cells in vitro. Methods Renal cell carcinoma ACHN and GRC-1 cells weretreated with different concentrations of matrine, and cell proliferation activity was detected by MTT assay to calculate thehalf inhibitory concentration. Cells treated with a half inhibitory concentration of matrine or an Akt pathway inhibitor werenamed the matrine group and inhibitor group, respectively; cells treated with both were the combined group, and untreatedcells were used as controls. Cell proliferation was detected by MTT, apoptosis was measured by flow cytometry, levels ofreactive oxygen species (ROS) were examined by laser scanning confocal microscopy, and the expression of p38, p-p38,and cleaved caspase-3 was investigated by western blotting. Results Matrine can inhibit the proliferation of renal cancercells in a concentration-dependent manner. Compared with the control group, the cell survival rate decreased, apoptosisrate, ROS level, p-p38 level and cleaved caspase-3 protein level increased in the matrine, inhibitor and combinationgroups. Compared with the matrine group and inhibitor group, the combined group had a lower survival rate, higherapoptosis rate, higher ROS level, higher p-p38 level and cleaved caspase-3 protein level. Conclusions Matrine combinedwith an Akt signaling pathway inhibitor inhibits the proliferation of renal carcinoma cells, and promotes their apoptosis. ROS and p38 signaling pathways may be one of the mechanisms of action of this effect.

Abstract:Objective To evaluate whether high-fat diet-induced obesity affects ovarian function in rats and tocontribute to an understanding of the mechanisms involved. Methods Female 5-week-old Wistar rats were fed a high-fatdiet (HFD group, n =40) or a normal diet (control group, n = 40). After 2 months of feeding, the estrous cycle and thenumber of cells in each stage of rat follicular development were evaluated. The levels of serum progesterone and estradiol ineach group were also measured. Male and female rats (1 ∶2) were then mated over a period of 1 week. Results HFDinduced obesity and hyperglycemia, without altering levels of serum triglycerides, cholesterol, or C-reactive protein. HFDalso altered ovarian function by prolonging diestrous phases, decreasing serum estradiol concentrations, and increasing thenumber of antral atretic follicles. Moreover, the HFD group developed follicular cysts and became pregnant later thancontrol group rats. Conclusions HFD induces follicular cyst development and disturbs the ovulatory process, leading to delayed pregnancy.

Abstract:Objective To observe the inhibitory effect of oxymatrine (OMT) combined with all-trans retinoic acid(ATRA) on diethylnitrosamine (DEN)-induced hepatocellular carcinoma in rats, and to explore its mechanism of action.Methods A rat model of hepatocellular carcinoma was established by DEN induction for 18 weeks. Wistar rats wererandomly divided into control, model, and intervention groups. Rats were killed at 6, 12, and 18 weeks after cancerinduction. Pathological changes of the liver tissues were observed by hematoxylin and eosin (HE) staining, related indexesof rat liver function were detected, and interleukin (IL)-6 expression was detected by enzyme-linked immunosorbent assay.The mRNA expressions of let-7a, nuclear factor (NF)-κB-p65, IL-6, and signal transducer and activator of transcription(STAT)3 were detected by quantitative real-time (RT-q)PCR, while the changes of STAT3 and p-STAT3 proteins weredetected by western blotting. Results The pathological examination revealed a significantly decreased number of livercancer nodules in the intervention group compared with the model group ( P <0. 05), while levels of serum IL-6, alanineaminotransferase(ALT), glutamyl transpeptidase(GGT), aspartate aminotransferase(AST), and alkaline phosphatase(ALP) were significantly lower ( P < 0. 05). RT-qPCR results showed that the expression of let-7a was significantlyincreased in the intervention group compared with the model group, while that of NF-κB-p65, IL-6, and STAT3 wassignificantly decreased ( P <0. 05). At the 18th week of the experiment, the expressions of STAT3 and p-STAT3 in theintervention group were significantly lower than in the model group ( P <0. 05). Conclusions OMT combined with ATRA can significantly inhibit DEN-induced hepatocarcinogenesis in rats and delay the growth of tumors to some extent.

Abstract:Cytochrome P450 2E1 ( CYP2E1) is an enzyme that can metabolize endogenous or exogenoussubstances, mainly in the endoplasmic reticulum. CYP2E1 is involved in the oxidative metabolism of exogenous chemicalssuch as steroids, fatty acids, prostaglandins, drugs, carcinogens and environmental pollutants. In alcoholic liver disease,nonalcoholic liver disease, cardiovascular diseases, and AIDS, and in the pathological process of diseases such as diabetesand Parkinson’s disease, the CYP2E1 expression levels rise, produce reactive oxygen species and reactive metabolites,cause oxidative stress, and deactivate DNA, protein and lipids, causing organ damage in the body. Therefore, targetedinhibition of CYP2E1 can be used to treat various diseases. Diallyl sulfide (DAS), an effective component in garlic, is aselective inhibitor of CYP2E1 and can reduce ROS, oxidative stress and apoptosis by inhibiting CYP2E1. This article reviews the potential role of DAS in targeted inhibition of CYP2E1 for treating related diseases.

Abstract:Small intestinal submucosa (SIS) treated via decellularization technology is a natural extracellularmatrix material. Because of its low immunogenicity and good biocompatibility, decellularized SIS is widely used for tissueregeneration and repair. However, the biological characteristics of SIS must be accurately understood to better transform andapply SIS in regenerative medicine. The function of SIS as a tissue-repair material depends on its composition andmechanical properties, the small molecules that the SIS degrades after being implanted, and the various properties andfunctions of these degraded molecules. This article reviews the physical and chemical composition, characteristics,processing method and applications of SIS to help understand the structure and function of SIS and promote its transformation and application.

Abstract:3-methyladenine (3-MA) is a classic autophagy inhibitor that mainly inhibits autophagosome formationand development. In the nervous system, 3-MA plays an important role in central nervous system diseases, peripheralnervous system diseases, nerve injuries, and neurodegenerative diseases. However, the exact effect of 3-MA in nervoussystem diseases is controversial, and its application lacks systematic discussion. This paper reviews both the role of 3-MA innervous system diseases and the relevant research on this role, and to provide new ideas for further research.

Abstract:Stroke is characterized by high mortality, disability, and recurrence rate, and is a disease withunsatisfactory therapies. Hemorrhage and ischemic strokes are the two main types of stroke, with the latter accounting for80% of all cases. Research has been conducted on the development of effective drugs to prevent and treat stroke, and theestablishment of a convenient and practical model is an important part of pathologic mechanism research and the drugscreening process. Among ischemic stroke models, the photothrombotic stroke model is increasingly used because of itsunique advantages such as its simple operation, very low mortality rate, no requirement for craniotomy, and the fact that itis less traumatic than other models. It can adjust the position of the region of the brain where the laser light strikes to controlthe location of blood clots, change the dosage of the photosensitizing dye, and the intensity and time of laser irradiation toaffect the size of focal ischemic formation. It is also suitable for research into the cellular and molecular mechanisms ofneurodegeneration, neuroprotection, and neurogenesis during ischemic stroke, and the development of anti-stroke agents.This paper summarizes the mechanisms, advantages, and disadvantages of the photothrombotic stroke model and its applications in drug development.

Abstract:Parkinson’ s disease is a degenerative disease of the central nervous system in which dopaminergicneurons in the substantia nigra gradually degenerate and are lost, leading to damage of the substantia nigra-striatumpathway. The main clinical treatments for this disease are drugs and surgery, but both only relieve short-term symptoms andhave many side effects. Moreover, neither results in a cure. Stem cells have the capacity for self-renewal and a multidirectionaldifferentiation potential. Stem cell transplantation can result in nerve regeneration and restoration within thedamaged tissue, bringing hope for a possible treatment for Parkinson’ s disease. This article compares and analyzes theseparation methods, modes of transplantation, efficacy, and advantages and disadvantages of different types of stem cells inthe treatment of Parkinson’s disease. It also summarizes possible mechanisms by which stem cells could provide a treatmentfor Parkinson’s disease. This article should aid the selection of cell types and methods of transplantation for use in clinical trials.