Abstract:Objective To investigate the localization method of microinjection of the solitary nucleus in rats using different parameters and to improve its success rate and accuracy. Methods Twenty rats were divided into a vertical injection group, a Bregma-based localization group, and a Lambda-based localization group according to the experimental progress. The injection point of the vertical injection group was 0. 5 mm at the anterior edge of the skull base of the rat, with a needle insertion depth of 7. 5 mm. The Bregma positioning group used Bregma-11. 0 mm and 0. 5 - 0. 7 mm beside the bilateral midline as the puncture point. The oblique insertion angle was 24° backwards and the depth was 8. 6 mm at the injection point. The Lambda positioning group used Lambda-3. 2 mm and 0. 5 - 0. 7 mm beside the bilateral midline as the puncture point. The oblique insertion angle was 24° backwards and depth was 9. 6 mm at the injection point. The positions of the fluorescent markers in the coronal section of the whole brain after injection of each group of rats were observed. Results The fluorescent label in the vertical injection group was located in the choroid plexus. The fluorescent labeling was unstable in the Bregma-based localization group, and was located in the coronal sections of the cerebellum, superficial brainstem, and choroid plexus. In the Lambda-based localization group, the fluorescent label position in the rats weighing approximately 300 g was located in the space between the cerebellum and the brainstem, while the rats weighing around 400 g could be accurately labeled in the solitary nucleus. Conclusions In rats of approximately 400 g, oblique puncture at a certain angle based on Lambda as the reference point can enable accurate injection into the nucleus of the solitary tract in rats.

Abstract:Objective To establish synovial cell primary cultures and a lipopolysaccharide ( LPS)-induced fibroblast-like synoviocyte ( FLS) inflammatory model of a normal knee joint in Sprague-Dawley ( SD) rats. Methods Normal knee joint synovium (80-120 g) was separated from specific-pathogen-free SD rats in an ice bath with phosphate- buffered saline. The primary synovium was cultured to the 3rd generation. Vimentin expression and FLS proliferation were detected via immunohistochemical staining and the 5-ethynyl-2′-deoxyuridine method. Synovial tissue was used as a control to detect characteristic protein expressions in the FLS in the 3rd-8th generations of primary culturing and to screen the FLS for high purity and physiological functions for use in subsequent experiments. After LPS-induced FLS inflammation, the mRNA and protein expressions of IL-1β and TNF-α were detected at different time points to determine the time point at which LPS could successfully induce an FLS inflammatory model. Finally, the cytokine expression, proliferative functions and characteristic proteins of the FLS before and after LPS induction were detected to provide experimental data to evaluate the FLS inflammatory model. Results FLS primary cells were successfully cultured with 0. 2% collagenase I digestion. The FLS purity in the 3rd generation exceeded 98% for detecting vimentin. By detecting characteristic FLS proteins, FLS with physiological functions that could be used for subsequent experiments were selected as the primary cells from the 3rd-7th generations. Analyzing the cytokines and characteristic proteins in FLS before and after LPS induction revealed that 1 μg / mL LPS is required to stimulate FLS cells for 3 h to replicate the FLS inflammatory model. Conclusion The LPS- induced FLS inflammatory model can be used to study inflammatory arthritis in vitro.

Abstract:Objective To establish a mouse model of endometriosis with fibrosis and to study its application. Methods A mouse model of endometriosis with fibrosis was established by modified intraperitoneal injection using BALB/ c mice; the peritoneal lavage fluid of mice was collected and the level of TGF-β1 in lavage fluid was detected by ELISA. The degree of abdominal adhesion was scored and the success of the endometriosis model was verified by hematoxylin-eosin(HE)staining, while the collagen deposition in the ectopic focus was detected by Masson staining and the degree of collagen deposition was evaluated. The expression of fibrosis-related proteins E-cadherin, collagen I, α-SMA, and smooth muscle myosin heavy chain II ( SMMHC-II) were detected by immunohistochemistry, and the degree of ectopic fibrosis was evaluated. Results The success rate of a mouse model of endometriosis with fibrosis constructed by a modified intraperitoneal injection method could reach 100%. The level of TGF-β1 in the peritoneal lavage fluid of mice increased significantly with the extension of modeling time. Masson staining and immunohistochemical staining of fibrosis-related proteins revealed a fibrotic phenotype in the abdominal cavity of endometriosis mice. Conclusions The improved intraperitoneal injection method has a high success rate in the establishment of a mouse model of endometriosis with fibrosis. TGF-β1 levels in peritoneal lavage fluid, Masson staining, and immunohistochemical staining of fibrosis-related proteins can evaluate the degree of pelvic fibrosis in mice with endometriosis.

Abstract:Objective This study compared mouse and rhesus monkey models of Graves’ disease ( GD), particularly regarding the immunological mechanisms, and provided research tools for further novel treatment. Methods Six-week-old female BALB/ c mice were injected intramuscularly with adenovirus expressing the A-subunit of thyrotropin receptor ( TSHR) ( A-sub-Ad) three times at 3-week intervals. The animals were euthanized 4 weeks after the final injection to obtain their blood, spleen cells, thyroid glands and other organs. Three-year-old female rhesus monkeys were injected intramuscularly with A-sub-Ad five times at 3-week intervals, and sera were prepared from blood samples collected at several time points during the immunization regimen. The animals were euthanized 4 weeks after the final injection to obtain their blood, spleen cells, thyroid glands and other organs. Serum total thyroxine ( TT4) and thyrotropin receptor antibodies (TRAb) levels and thyroid morphology were detected in both the mice and rhesus monkeys to identify the thyroid function. The body weights of the mice and rhesus monkeys were recorded during the experiment as well as the resting heart rate of the rhesus monkeys. Flow cytometry was performed to detect the changes in splenic CD4 + CD25 + Foxp3 + cell proportions. Results After the final immunization, compared with the controls, the A-sub-Ad-injected mice developed significantly higher TRAb and TT4 levels [(8. 1 ± 0. 6) IU/ I vs ( 423. 1 ± 61. 4) IU/ I and ( 57. 1 ± 2. 9) μg / dL vs (96. 7 ± 13. 8) μg / dL, respectively, P< 0. 05]. Using the mean ± 2 standard deviations as the normal range, all A-sub- Ad-injected mice and rhesus monkeys showed positive TRAb levels, and 75% of the mice and half of the rhesus monkeys had increased TT4 levels. The thyroid pathology confirmed the GD-induced changes in the mice and rhesus monkeys. Different from the thyroid follicular epithelial cells in the controls, which were lowly cubic or flat, 6 / 8 mice and 3 / 6 rhesus monkeys in the model group showed obvious follicular epithelial hyperplasia with cubic or tall columnar-shaped cells, and papillary structures caused by hyperplasia protruded into the follicular cavity in part of the visual field. However, no lymphocyte infiltration occurred in the thyroids of either the mice or rhesus monkeys. In addition, the rhesus monkeys with GD lost body weight and had significantly increased resting heart rates (P< 0. 05). Flow cytometry showed decreased CD4+ CD25+Foxp3+ cell proportions in both the mouse and rhesus monkey GD groups compared with those of the control groups. Conclusions Compared with the GD rhesus monkey model, the GD mouse model showed a shorter induction time and a higher GD incidence. However, in terms of physiology and immune response, especially with GD complications, GD rhesus monkeys showed more similarities to GD patients. In future research to explore the pathogenesis and evaluate new treatments for GD, scholars can choose the appropriate research tools according to the different characteristics of the two GD animal models.

Abstract:Objective To explore the effect of EtOAc extract from Saxifraga taugutica (EST) on carcinoma cells and orthotopic-transplanted tumors. Methods Murine cancer cells ( H22 cells ) were incubated with different concentrations of EST for 48 h in vitro. Morphological changes were examined by laser confocal microscopy after AO/ EB double staining. Apoptosis of carcinoma cells was determined by DNA electrophoresis. and flow cytometry. Orthotopic- transplanted tumors in mice were established in vivo. The tumor inhibition rate, thymus index, spleen index, and liver pathology were compared between low-dose, medium-dose, and high-dose groups. Results Apoptotic morphological changes of H22 cells were observed, with the level of apoptosis gradually increasing with increased EST concentration. DNA electrophoresis demonstrated a typical ladder pattern. Flow cytometry revealed that the apoptosis rate of H22 cells was 31% in the EST group with 500 μg / mL. In vivo experiments showed that the tumor inhibition rate was dose-dependent to some extent; the best tumor inhibition rate was 64. 3% in the high dose group, followed by tumor inhibition rates of 46. 7% in the medium dose group and 33% in the low dose group. Spleen and thymus indexes in the EST group was significantly higher than those in the model group (P< 0. 05). Histological observation showed that the proliferation and invasion of cancer cells was inhibited to different degrees, and karyopyknosis of cancer cells was identified in the EST group. Conclusion EST could enhance the immune function of mice, induce apoptosis of tumor cells, and significantly inhibit the proliferation of carcinoma cells in a dose-dependent manner.

Abstract:Objective To explore the therapeutic effect and mechanism of Qingfei Touxie decoction on mice with Mycoplasma pneumoniae (MP). Methods Forty male specific-pathogen-free BALB/ c mice were randomly divided into the control, model, low-dose, medium-dose, and high-dose Qingfei Touxie decoction groups, with n = 8 mice per group. A mouse model of MPP was established via nasal instillation of solution containing MP. Mice in the model and control groups received 0. 9% saline solution by oral gavage, and mice in the Qingfei Touxie decoction (QFTXT)-treated groups received QFTXT at low (200 mg / kg body weight), medium (400 mg / kg body weight) and high (800 mg / kg body weight) doses daily. After 2 weeks of treatment, the lung tissue indices of the mice were calculated, and the pathological changes in the lung tissue were observed via hematoxylin and eosin staining. Apoptosis in the mouse lung cells was detected via TUNEL staining, and the interleukin (IL)-1β, TNF-α, IL-6, and IL-18 levels were detected using enzyme-linked immunosorbent assays. NLRP3, MyD88 and NF-κB mRNA expressions in the lung tissue were detected using qPCR, and the NLRP3, MyD88 and p-NF-κB protein expressions in the lung tissue were detected via western blot. Results Qingfei Touxie decoction significantly reduced the lung indices of the MPP mice and reduced the lung pathological damage and apoptotic rates (P< 0. 05). Qingfei Touxie decoction also significantly reduced the IL-1β, TNF-α, IL-6 and IL-18 levels ( P< 0. 05) and NLRP3, MyD88 and NF-κB mRNA and protein expressions ( P< 0. 05). Conclusions Qingfei Touxie decoction attenuated lung injuries in mice with MP pneumonia, possibly by inhibiting NLRP3 inflammatory bodies and the NF-κB signaling pathway.

Abstract:Objective To elucidate the onset process and pathological characteristics of spontaneous hindlimb paralysis in a hyperlipidemia-susceptible (WSHc) rat population, and to preliminarily study the pathogenesis mechanism. Methods In a WSHc rat population, eight spontaneous hindlimb paralysis rats and eight age-matched rats without paralysis symptoms from the same family were fed with normal chow or high-fat chow to examine their susceptibility to hyperlipidemia. Magnetic resonance imaging and histopathology were used to examine central neuropathy in different parts of hindlimb paralysis rats. TUNEL immunohistochemistry was used to detect apoptosis. The mRNA expression of Caspase-1 and IL-1β in the central nervous system was determined by RT-PCR, and the corresponding protein expression was determined by Western Blot. Results Both male and female WSHc rats with hindlimb paralysis developed the paralysis. The sensitivity to high-fat diet was not significantly different between WSHc rats with and without hindlimb paralysis. No significant lesions were observed in the brain and cerebellum by magnetic resonance imaging. Histopathology showed a large amount of inflammatory cell infiltration and TUNEL-positive staining in the middle and posterior spinal cord of hindlimb paralysis WSHc rats. Compared with the non-hindlimb paralysis WSHc rats, the mRNA and protein expression of Caspase-1 and IL- 1β in the middle and posterior spinal cord was significantly increased (P < 0. 05, P< 0. 01) in the hindlimb paralysis WSHc rats. Conclusions Spontaneous hindlimb paralysis of hyperlipidemia-susceptible WSHc rats is a progressive lesion, which occurs in the middle and posterior spinal cord and is pathologically featured by inflammatory cell infiltration, neuronal degeneration and apoptosis. This pathogenesis may be related to excessive activation of caspase-1.

Abstract:Objective To screen potential biomarkers to explore the pathogenesis in a rat model of isoproterenol- induced cardiac hypertrophy. Methods Isoproterenol 30 mg / ( kg·d) was used to establish the rat model of cardiac hypertrophy via intraperitoneal injection for 14 consecutive days. The rat cardiac hypertrophy model was evaluated via the cardiac index. Ultra high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF- MS) was conducted to detect serum metabolites in normal and model rats. MPP software was used to analyze metabolic differences. The Human Metabolome Database (HMDB) was used to identify biomarkers. MetaboAnalyst 4. 0 was used to analyze metabolic pathways. Results Serum metabolites in the rats with isoproterenol-induced cardiac hypertrophy differed significantly from those of the normal rats, and 10 potential biomarkers were identified. Compared with the normal group, sphingosine 1-phosphate and dihomo-γ-linolenic acid in the model group were significantly downregulated, and D-fructose- 1,6-bisphosphate, deoxyadenosine, N-acetylmethionine, phytosphingosine, allantoin, 3-keto-β-D-galactose, octane, and glycerol were significantly upregulated. Conclusions The metabolic pathways involved in isoproterenol-induced cardiac hypertrophy include sphingolipid metabolism, glycerolipid metabolism, galactose metabolism, biosynthesis of unsaturated fatty acids, and purine metabolism. This study provides a basis for understanding the metabolic changes in the circulating blood in a model of isoproterenol-induced cardiac hypertrophy.

Abstract:Objective To observe the effects of electroacupuncture (EA) on ipsilateral paw withdrawal thresholds (PWTs), activation of satellite glial cells, and the P2X7 receptor in the dorsal root ganglia (DRG) of rats with chronic inflammatory pain, and further explore the peripheral mechanism of EA against chronic inflammatory pain. Methods Part 1: Rats were randomly divided into the control group (Con group; n = 12), CFA-induced inflammatory pain group (CFA group; n = 12), CFA model plus EA treatment group (CFA + EA group; n = 12), and CFA model plus sham EA treatment group (CFA + sEA group; n = 12). The CFA inflammatory pain model was established by injecting 0. 1 mL CFA into the plantar of the rat right hind foot. EA at a 2 / 100 Hz alternative frequency and gradual intensity (0. 5, 1, and 1. 5 mA, 10 min each) was administered at acupoints “Zusanli” and “Kunlun” for 30 min per day for 7 days. PWTs were detected at 1, 3, 7, 8, 10, 12, and 14 days after CFA injection. Protein expression of glial fibrillary acidic protein (GFAP) and P2X7 receptor in DRG at day 14 post-CFA was measured by western blotting. Part 2: CFA rats were randomly divided into the CFA plus DMSO group (CFA + DMSO group; n = 8), CFA plus P2X7R agonist A740003 group (CFA + A740003 group; n = 10), EA plus normal saline group (CFA + EA + NS group; n = 8), and EA plus P2X7R agonist BzATP group (CFA + EA + BzATP group; n = 12), which were intrathecally administrated with DMSO, A740003, normal saline, and BzATP respectively. PWTs were measured at pre-surgery, pre-CFA, post-CFA, and post-intrathecal injection. Results Part 1: PWTs of CFA rats were decreased significantly (P < 0. 01) and EA significantly increased PWTs (P < 0. 01). CFA injection significantly increased the expression of GFAP and P2X7 receptor in the ipsilateral L4-6 DRG of rats (P< 0. 05). EA significantly reduced the expression of GFAP and P2X7 receptor in the L4-6 DRG of rats (P < 0. 05), but sham EA had no obvious effect (P > 0. 05). Part 2: Compared with the CFA + DMSO group, intrathecal injection of P2X7 receptor antagonist A740003 significantly increased PWTs of CFA rats (P < 0. 01). PWTs in the CFA + EA + BzATP group were significantly lower than those in the CFA + EA + NS group (P < 0. 01). Conclusions Electroacupuncture alleviates chronic inflammatory pain in rats, which might be related to the reduction of satellite glial cell activation and decrease of P2X7 receptor activation in the rat DRG. Inhibition of satellite glial cell activation and a decrease of P2X7 receptor expression contribute to EA treating chronic inflammatory pain, which may be one of the peripheral mechanisms of EA analgesia.

Abstract:Objective To assess visceral hypersensitivity and constancy of visceral hypersensitivity models of irritable bowel syndrome (IBS) in rats. Methods Two-day-old Sprague-Dawley rat pups were randomly divided into the neonatal maternal separation group (group A), the unpredictable neonatal maternal separation group (group B), and the control group ( group C). After successful modeling, the rats’ general conditions and stools were observed, and their visceral hypersensitivity and constancy were determined via the abdominal wall withdrawal reflex and visceral pain threshold. After testing, the rats’ colonic tissue was taken for pathological examination. Results Body weight and colonic pathological examinations did not differ between the groups (P> 0. 05). Compared with groups A and C, rats in group B had increased stools and stool water content. Compared with groups B and C, the abdominal withdrawal reflex(AWR) threshold was highest in the group A rats and peaked at 9 weeks of age. The visceral pain threshold did not differ between groups A and B but was lower in both groups compared with that of group C (P > 0. 05). Conclusions Both the neonatal maternal separation and the unpredictable neonatal maternal separation models simulated the IBS visceral hypersensitivity model. The unpredictable neonatal maternal separation model better simulated a diarrheal IBS model. The visceral hypersensitivity model was time-limited, and the optimal experimental time was within 9 weeks of age.

Abstract:Objective To establish an efficient rat model of hyperuricemia with liver and kidney injury, gouty arthritis and other complications. Methods Forty-eight male Sprague-Dawley rats were randomly divided into the blank control group (control group), oteracil potassium group (OA group), oteracil potassium with 6-hydroxypurine group (OA + H group), oteracil potassium with yeast extract group (OA + YE group), 10% fructose with 0. 2% ethambutol group (10% Fru + 0. 2% EMB group), and the 2% oteracil potassium mixed with 12% yeast extract peculiar feed group (OA PO group) (n = 8 rats per group). Blood was collected from the tail vein once weekly, the serum uric acid and urea values were detected, and the model was studied for 5 consecutive weeks. Results Compared with the control group, the OA PO group reached high serum uric acid levels with severe liver and kidney damage and joint inflammation in week 5. The uric acid values of the OA + H, OA + YE and 10% Fru + 0. 2% EMB groups failed to reach high levels. However, biochemical indicators showed that liver and kidney damage occurred in these four groups. Hematoxylin and eosin staining showed that the knee joints of the model group had synovial thickening and scattered and disordered arrangement to varying degrees. Conclusions The 2% oteracil potassium mixed with 12% yeast extract diet is the best diet for establishing a rat model of hyperuricemia and its complications and is suitable for studying hyperuricemia-induced renal injury and arthritis.

Abstract:Objective To explore the effects of purslane, licorice, and dandelion compound on insulin resistance in type 2 diabetic rats. Methods Low-dose streptozotocin was combined with a high-sugar and high-fat diet to establish type 2 diabetic rats. After successful modeling, they were randomly divided into control ( normal), model, purslane, licorice, dandelion, compound, and metformin groups. After 4 weeks of intervention, Fasting blood glucose (FBG), Serum total cholesterol ( TC), Triglyceride ( TG), High-density lipoprotein ( HDL), Low-density lipoprotein ( LDL), and Fasting insulin (FINS) were measured in each group of rats. The changes in the values were calculated and Atherosclerosis index (AI), Insulin sensitivity index ( ISI), and Insulin resistance index ( IRI) values were calculated. The pancreatic tissue was cut into sections and the morphological changes were observed after staining. Results (1) The weight of each intervention group was significantly different from that of the model group (P< 0. 01), but the effect was not as good as metformin. (2)At 2 weeks, the FBG value of each intervention group was significantly lower than that of the model group (P< 0. 05); at 4 weeks, the FBG value of each intervention group was significantly lower (P< 0. 01), and there was no statistical significance between the compound group and the metformin group before and after 2 weeks and 4 weeks of intervention (P > 0. 05). ( 3) The 0. 5, 1, and 2 h blood sugar values of the compound and metformin groups were significantly lower after the glucose load, but there was a significant difference in 2-h blood glucose (P< 0. 05).(4) The ISI and IRI values of each intervention group were significantly higher and lower than those of the model group, respectively (P< 0. 01). (5) The levels of TC, TG, HDL-C, and LDL-C in the metformin group were similar to those in the compound group (P> 0. 05). (6)Microscopic observation revealed that the compound group underwent repair and proliferation, and the cells were arranged evenly and uniformly in size, similar to those in the metformin group. Conclusions Purslane, licorice, and dandelion compound can regulate glucose and lipid metabolism disorders, repair islet β cells, and thereby improve insulin resistance. The overall effect of combination treatment is better than that achieved with single treatment.

Abstract:Objective A rat model of vaginitis was established to provide a model for developing and evaluating therapeutic drugs for vaginitis. Methods Gardnerella vaginalis, Streptococcus albus and Group B Streptococcus were isolated and purified from clinical vaginal samples. Twenty-six female rats were treated with 20 mg / kg estradiol benzoate and 10 μg / kg streptomycin in the morning, and mixed strains were inoculated into the vagina in the afternoon. Ten female rats serving as the control group were subcutaneously administered 20 mg / kg estradiol benzoate once daily for 3 days. Mixed bacterial infections in the vaginas of the rats were observed by collecting vaginal discharge and lavage fluid, then the rats were grouped according to body weight and strain load. Twenty rats were randomly divided into either the model group (54 mg / day compound metronidazole vaginal suppository once daily for 5 days) or the drug validation group, with 10 animals per group. The control and model groups were injected vaginally with 0. 9% sodium chloride. After the last drug administration, the bacterial strain loads were detected via vaginal lavage, and the vaginal smears, cleanliness and pH were examined from the vaginal discharge. The vaginal tissue was performed to observe the histomorphological changes by HE staining. Results In the model group, the vaginal walls became thickened, the stratified squamous epithelial cells became denatured and necrotic, and the lamina propria became engorged and edematous. Neutrophils were the primary cells, and lymphocyte and monocyte infiltration occurred, with scattered punctiform hemorrhaging, obvious fibroblast proliferation in the mastoid region, and slight reticular fiber proliferation. The mixed bacterial numbers, pH values and cleanliness scores were significantly higher in the model group than in the normal group (P < 0. 01), and the negative conversion rate was significantly reduced ( P < 0. 01). A compound metronidazole vaginal suppository significantly reduced the numbers of mixed bacteria, pH values and cleanliness scores; increased the negative rate for vaginal bacteria and significantly attenuated the degree of the vaginal lesions. Conclusions After pretreatment with estrogen and streptomycin sulfate injections, a rat model of mixed vaginitis was successfully established by inoculating mixed bacteria, thus providing an effective animal model for drug screening and evaluation of female reproductive system.

Abstract:Objective To observe whether hFIX is stably expressed in adipose-derived stem cells (ADSCs) and to explore whether ADSCs can be used as a vector cell line for hemophilia gene therapy following transformation by recombinant adenoviruses carrying human coagulation factor IX ( hFIX) gene, F9. Methods ADSCs were isolated from C57BL/ 6 mice and cultured by tissue mass suspension. The third generation of ADSCs was transfected by recombinant adenovirus carrying F9 and GFP fluorescence marker. After transfection, the cells were subcultured again to the 4th and 5th passages. The level of fluorescence expressed by the cells was observed by fluorescence microscopy, the expression of F9 was detected by RT-PCR, and the protein expression of hFIX was detected by ELISA and western blotting. Results Fluorescence was observed after recombinant adenoviral transfer, and was still identified after cell passaging. RT-PCR revealed that all four groups of ADSCs could express internal reference GAPDH, but there was no expression of the target gene F9 in group A, and the expression of F9 could be detected in group D. Detection of hFIX:Ag by ELISA showed that the detection values of groups A (81. 62 ± 8. 82) ng / mL, B (52. 50 ± 3. 25) ng / mL, and C (47. 41 ± 4. 00) ng / mL were significantly higher than that of group D (0. 76 ± 0. 44) ng / mL, and there was a significant difference between the two groups (P< 0. 05). hFIX expression in the four groups of ADSCs was detected by western blotting. The result showed that the gray value of histone expression in groups A (0. 68 ± 0. 10), B (0. 49 ± 0. 15) and C was significantly higher than that in group D (0. 02 ± 0. 01), and the difference was statistically significant (P< 0. 05), while the gray value of histone expression in group A was significantly higher than that in group D (P< 0. 05). Conclusions The recombinant adenoviruses were transformed into ADSCs and cultured after passaging, and could still express high hFIX activity, thus demonstrating their potential as vector cells for hemophilia gene therapy.

Abstract:Objective Artificially formulated milk fed after birth is essential for cultivating germ-free rats. Lactose-free milk powder with deep hydrolyzed protein is easily absorbed by infants and reduces indigestion and allergic reactions. We detected the biochemical indexes of germ-free rats to investigate whether adding lactose-free milk powder to their milk would have a positive effect on the rats. Methods Pregnant Sprague-Dawley rats were determined to be in labor. After surgically removing the uterus, the young rats were housed in isolators and artificially lactated for 22 days. Body weight and survival rates were recorded. The experimental group received lactose-free milk powder with deep hydrolyzed protein in their milk, and the control group received regular formula milk. The blood biochemical indexes were measured at 8 weeks. Results During the first 14 days of artificial feeding, body weight and survival rates did not differ between the experimental and control groups. The weights and survival rates of the experimental group began to increase compared with those of the control group on day 14. The survival rate and body weights of the experimental group were significantly higher than those of the control group on day 22 [ 37. 18% vs 17. 78% and ( 9. 96 ± 0. 49) g vs ( 13. 36 ± 0. 59) g, respectively]. Blood biochemical indexes of the germ-free rats showed that their aspartate aminotransferase levels decreased, and their glucose levels increased. Conclusions In cultivating germ-free rat pups, lactose-free milk powder with deep hydrolyzed protein can be added to formula milk to effectively increase the weight of the young rats and reduce their mortality rate. Adding lactose-free milk powder to formula milk substantially changed the blood biochemical indexes of female germ-free rats.

Abstract:Rotator cuff tendon injury is the main cause of acute and chronic shoulder pain and a limited range of motion. Surgical intervention is usually needed in cases of acute injury with full-thickness tears. The difficulty in repairing rotator cuff injuries lies in reconstructing the bone-tendon interface. Although various animal models of rotator cuff injuries induced by surgical trauma have been developed, suitable animal models and exact treatment method are lacking. This paper summarizes the establishment method and applicable scope of rotator cuff tendon injury-induced animal models and explores the selection of animal models and functional evaluation method to provide a corresponding theoretical basis for basic scientific research on rotator cuff injuries.

Abstract:Acute spinal cord injury is a central nervous system injury with a high disability rate and serious consequences, which brings a heavy burden to individuals, families and society. No effective treatments exist for spinal cord injuries, but different therapies, including stem cells, drugs and tissue engineering have been used in various animal models with some success. Similar questions remain as to whether the mechanism of injury, effectiveness, and integrity of animal models can mimic what is seen clinically. This paper describes the application, advantages, and disadvantages of several common animal models of acute mechanical spinal cord injury, as well as the selection of common animal types and injury segments, to provide references for selecting more effective animal models.

Abstract:The metastatic model of transplanted lung cancer in mouse is an important tool in lung cancer research, and has great significance in interpreting the metastasis mechanism of lung cancer and in studying drugs related to lung cancer. In this paper, the frequent sites of lung cancer metastasis and the species differences of tumors were comprehensively considered, and this model was further classified, while the modeling method and evaluation index of each type were expounded in detail to provide model references for lung cancer-related research.

Abstract:Chronic obstructive pulmonary disease ( COPD) is characterized by incomplete reversible persistent progressive airflow limitations, and always presents acute exacerbations and serious complications. Establishing standardized animal models that are consistent with the clinical COPD characteristics of COPD is important for exploring its development, and morphological changes, as well as new therapeutic approaches. new therapeutic approaches. Many studies have been conducted on COPD, but there is no best method exists for constructing animal models of COPD. Here, we describe and compare the types of animals used to establish the COPD models and the advantages, and disadvantages, and clinical applications of each model.

Abstract:Food allergies are rapidly becoming a global health problem. Food allergic reactions are caused by Th2 responses to harmless foods, leading to IgE-mediated mast cell degranulation and related inflammatory responses. Food allergy murine models are helpful for studying the pathology, physiology and new treatment strategies for human food allergies. These models can be divided into adjuvant-dependent, adjuvant-free, genetic-engineered and humanized mouse models. However, no animal models can reproduce the entire range of human pathological features; therefore, specific mouse models of food allergies must be used to address specific food allergy research issues. This article summarizes the literature, research progress on modeling method, and advantages and disadvantages of different murine models of food allergies over the last 20 years and provides a reference and scientific basis for researching food allergy mechanisms and developing new treatment method .

Abstract:Tumor necrosis factor (TNF) is essential for the development and control of tuberculosis. The zebrafish- marine mycobacterium model is one of a growing number of models currently used in tuberculosis research. This model can help us to explore the relationship between TNF and macrophages, granuloma, or mycobacterium, so as to better understand the relevant factors affecting TNF production and the role of TNF in the course of tuberculosis.