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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
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ObjectiveTo determine the therapeutic effect of Gegentang granules on a disease-syndrome mouse model combining human coronavirus 229E （hCoV-229E） pneumonia with Hanshi Yidu Xifei syndrome in vivo. MethodMice were randomly divided into normal group， infection group， cold-dampness group， model group， chloroquine phosphate group （0.18 g·kg-1）， interferon-α2b （IFN-α2b） group （1.83×106 U·kg-1）， Gegentang granules high-dose and low-dose groups （6.6， 3.3 g·kg-1） with 10 mice in each group. Cold-dampness environment and hCoV-229E infection were used for modeling， and the general status and lung index of mice in each group were observed. The viral load in lung tissue was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， the pathological changes in lung tissue were evaluated by hematoxylin-eosin （HE） staining， the levels of serum gastrointestinal hormones and inflammatory factors in lung tissue were detected by enzyme-linked immunosorbent assay （ELISA）， and the percentage of peripheral blood lymphocytes was detected by flow cytometry. ResultComparing with model group， Gegentang granules could significantly alleviate the physical signs of Hanshi Yidu Xifei syndrome， including listlessness， weakness of limbs， sticky stool， etc. Comparing with model group， Gegentang granules high-dose group significantly reduced lung index， histopathological score of interstitial lung and bronchus， and the level of serum motilin （P<0.05， P<0.01）， two doses of Gegentang granules could significantly increase the level of serum gastrin （P<0.05， P<0.01）， the percentage of CD4+， CD8+ T lymphocytes in peripheral blood （P<0.05， P<0.01）， and the level of tumor necrosis factor-α （TNF-α） in lung tissue was significantly decreased （P<0.01）， and the level of interleukin-6 （IL-6） showed decreasing tendency. ConclusionGegentang granules has therapeutic effect on model mice. It can improve the appearance and behavior characterization， regulate the level of gastrointestinal hormones， decrease lung index and histopathological score， and possibly play an immunomodulatory role by inhibiting the expression of inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes.
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BACKGROUND: Type l diabetes mellitus is a T cell-mediated autoimmune disease resulting in pancreatic islet cell damage. In this study, immunotherapy was used to deal with type l diabetes mellitus and stem cell transplantation was used to repair damaged islet p cells, attempting to explore a new treatment for type l diabetes mellitus. OBJECTIVE: To observe the efficacy of human umbilical cord mesenchymal stem cells combined with immunotherapy for the treatment of type l diabetic mice. METHODS: Fifty BALB/c Foxp3-DTR-EGFP positive mice were selected, six of which were randomly selected as normal control group and the remaining of which were intraperitoneally injected with streptozotocin and diphtheria toxin to prepare an animal model of type l diabetes mellitus. After successful modeling, randomization was performed in model mice and there were four groups: model group (normal saline), immunotherapy group (subcutaneous injection of dexamethasone (10 μg) and insulin (10 μg) mixture), cell transplantation group (injection of human umbilical cord mesenchymal stem cells (1 X106 cells per mouse) through the tail vein, and combined treatment group (the combination of immunotherapy and cell transplantation as described above). At 4 weeks after treatment, changes in blood glucose, C-peptide, body mass, pancreatic histopathology and insulin-positive area were observed in each group. RESULTS AND CONCLUSION: (1) Compared with the normal control group, the blood glucose level of the model group increased (P < 0.01) the C peptide level and body mass decreased (P < 0.01), and the islet was severely atrophied, with decreased number of islet cells and reduced insulin-positive area. (2) Compared with the model group, the blood glucose level of the immunotherapy group decreased (P > 0.05), the C-peptide level and body mass did not change significantly (P > 0.05), the islet cells increased in number, and the insulin-positive area increased. (3) Compared with the model group, the blood glucose level of the cell transplantation group and the combined treatment group decreased (P > 0.05), the C peptide level and body mass increased (P < 0.05), the islet cells increased in number, and the insulin-positive area increased. These findings reveal that either human umbilical cord mesenchymal stem cells or immunotherapy can improve the islet function of type l diabetic mice, and the combination treatment has better outcomes.
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BACKGROUND:Domestic and international studies have confirmed that human umbilical cord mesenchymal stem cel s could be induced to differentiate into islet-like cel s, but little is reported about the changes of insulin and nestin expressions during the differentiation phase. OBJECTIVE:To observe the changes of insulin and nestin expressions during the differentiation of human umbilical cord mesenchymal stem cel s into islet-like cel s. METHODS:Human umbilical cord mesenchymal stem cel s were cultured using UltraCULTURE medium in vitro. Stem cel s were cultured for three generations to observe cel morphological changes under an inverted microscope, to test immunophenotype by flow cytometry, and to identify the capacity of osteogenesis and adipogenic differentiation. Induction protocol was divided into two stages. In stage 1, stem cel s were induced for 14 days in the UltraCULTURE medium with 4 nmol/L activin A, 25μg/L epidermal growth factor, 100μg/Lβ-nerve growth factor, 10 mmol/L nicotinamide. In stage 2, the cel s were cultured in the UltraCULTURE medium with 1%insulin-transferin-selenium, 10 mmol/L nicotinamide, 10μg/L basic fibroblast growth factor for an additional 14 days. The expressions of nestin and insulin in those differentiated cel s were tested by flow cytometry, and zinc ion expression in the islet-like cel clusters was identified by dithizone staining. RESULTS AND CONCLUSION:During the differentiation process, the insulin level was increased gradual y in the induction group and reached a higher level on day 28, but the insulin expression showed negative in the control group. In addition, on day 14 of induced differentiation, the nestin expression reached the peak and then gradual y reduced along with the prolonged inductive time. On day 28 of induction, islet-like cel clusters formed and were positive for dithizone staining. In this experiment, the umbilical cord mesenchymal stem cel s were successful y induced and differentiated into islet-like cel s, accompanied with the variation of insulin and nestin expression.
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Abstract] Objective To investigate the in vivo therapeutic effects of an extract of herb medi-cines, YiGanQingDuKeLi, in combination with adefovir dipivoxil (ADV) on the rebound of duck hepatitis B virus ( DHBV) multiplication after withdrawal of ADV treatment.Methods Peking ducks were infected with DHBV positive serum samples for 7 days and then screened by SYBR Green real-time PCR.The ducks positive for DHBV were randomly divided into five groups including the model control group, the ADV treat-ment group, the herb treatment group, the high-dose combination therapy group and the low-dose combina-tion therapy group.The ducks in the ADV treatment and the herb treatment groups were respectively treated with distilled water and YiGanQingDuKeLi (1.2 g/ml) for 14 days after the treatment of ADV (0.25 mg/ml) for 21 days.The ducks in the high-dose group were treated with YiGanQingDuKeLi (1.2 g/ml) for 14 days after the combined treatment with high-dose YiGanQingDuKeLi (1.2 g/ml) and ADV (0.25 mg/ml) for 21 days.The ducks in the low-dose group were treated with YiGanQingDuKeLi (0.6 g/ml) for 14 days after the combined treatment with YiGanQingDuKeLi (0.6 g/ml) and ADV (0.125 mg/ml) for 21 days.Blood samples were collected from each duck via leg vein after 0, 7, 14 and 21 days of drug adminis-tration and after 7 and 14 days of drug withdrawal.The levels of DHBV-DNA, alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) in blood serum samples were detected.Results Compared with the model group, the levels of DHBV-DNA, ALT and AST in ducks from the herb treatment group and combined treatment groups were decreased before the discontinuation of ADV treatment ( P<0.05 or P<0.01).Moreover, the titers of DHBV-DNA in ducks treated with high doses of drugs were much lower than those from ADV treatment group.The levels of DHBV-DNA, ALT and AST in ducks treated with herb medi-cine and high doses of drugs remained at relatively low levels after the cessation of ADV treatment, but re-bounded significantly in ducks with ADV treatment.The levels of DHBV-DNA and ALT rebounded slightly in ducks treated with low doses of drugs as compared with those of ADV treatment group ( P<0.01 or P<0.05).Conclusion The treatment of YiGanQingDuKeLi in combination with ADV could inhibit not only the in vivo replication of DHBV, but also the rebound of DHBV multiplication after ADV withdrawal.
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Background:Acute hepatic failure( AHF)is a common pathophysiological process of end-stage liver disease with complex etiology,difficulty in diagnosis and high mortality rate. Aims:To investigate the protective effect of nicotinamide on AHF in mice. Methods:AHF model in mice was established by intraperitoneal injection with D-galactosamine 700 mg/kg and lipopolysaccharide 10 μg/kg. Fifty-four mice were divided into blank control group,nicotinamide control group, AHF model group and low,moderate,high dose(400,800,1 000 mg/kg)nicotinamide groups,levels of ALT,AST, TNF-α and IL-6 were determined,HE staining was used to examine hepatic histological injury,liver cell apoptosis was measured by TUNEL assay,and protein expression of Caspase-3 was detected by Western blotting. Another 40 mice were divided into AHF model group,saline group and low,moderate,high dose(400,800,1 000 mg/kg)nicotinamide groups,mortality rate was observed dynamically. Results:Compared with blank control group and nicotinamide control group,levels of ALT and AST were significantly increased(P<0. 05),infiltration of inflammatory cells and necrosis of cells and levels of TNF-α and IL-6 were significantly increased( P <0. 05 ),and apoptosis of liver cells and protein expression of Caspase-3 were significantly increased in AHF model group(P <0. 05). In groups pretreated with low, moderate and high dose nicotinamide,all the above-mentioned indices were significantly improved in a dose-dependent manner(P<0. 05). Survival rate in low,moderate,high dose nicotinamide groups was significantly higher than that in AHF model group(37. 5%,62. 5%,100% vs. 0%,P all <0. 05). Conclusions:Nicotinamide could protect mice from AHF via inhibiting inflammatory response and hepatocyte apoptosis,thereby increase the survival rate.
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BACKGROUND:At present, a great quantity of research has shown the effectiveness of traditional Chinese medicine and bone marrow mesenchymal stem cel s for vascular restenosis. However, studies concerning their combined application to restenosis after percutaneous transluminal angioplasty with diabetes mel itus are presently lacking. OBJECTIVE:To observe the effects of combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription on restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus. METHODS:A dog model of vascular restenosis with diabetes mel itus was established by bal oon injury of femoral artery and intravenous injection of al oxan. After successful model induction, 22 dog models were randomly divided into three groups:model group (n=6), treatment with Chinese medicine (n=8), and combined treatment with bone marrow mesenchymal stem cel s and Chinese medicine (n=8). Serum vascular endothelial growth factor levels were measured using enzyme-linked immunosorbent assay preoperatively and at 1, 2, 4 and 8 weeks postoperation. Samples of vessels were taken to conduct pathomorphological observation and quantitative analysis of proliferation degree. Tissues, including heart, liver, kidney and pancreatic gland, were col ected to evaluate the safety of stem cel transplantation using hematoxylin-eosin staining at 8 weeks postoperation. RESULTS AND CONCLUSION:Serum vascular endothelial growth factor levels began to increase at 1 week postoperation in the Chinese medicine group and combined treatment group, at 4 weeks postoperation in the model group compared with preoperation (P<0.05). At al time points, serum vascular endothelial growth factor levels were highest in the combined treatment group, but lowest in the model group (P<0.05). Quantitative analysis of vascular proliferation demonstrated that at 8 weeks postoperation, new intimal area, new intimal/medial areas and stenosis rate were highest in the model group, but lowest in the combined treatment group at 8 weeks postoperation (P<0.05). Safety assessment of stem cel transplantation showed morphological structures of the heart, liver, kidneys and pancreas were normal, no necrosis. In a word, the effects of the combined application of bone marrow mesenchymal stem cel s and benefiting-Qi nourishing-Yin and dissolving-congestion prescription were much pronounced in preventing restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus rather than single therapy of Chinese medicine. It is a safe and effective treatment to prevent vascular restenosis after percutaneous transluminal angioplasty in dogs with diabetes mel itus.
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BACKGROUND:Previous tudies have shown that the anti-aging effects of stem cel s with lycopene are more significant, and can also significantly improve the aging body immune function. OBJECTIVE:To investigate the anti-aging effects of human umbilical cord mesenchymal stem cel s combined with lycopene on the aging beagles. METHODS:Sixteen aging beagles (6-7 years old) were randomly divided into two groups:aging control group and aging treatment group;young beagles (3-4 years old) were chosen as young control group. In the aging treatment group, human umbilical cord mesenchymal stem cel s combined with lycopene was given;while in the other two group, an equal amount of DMEM/F12 cel culture medium and sunflower oil was given. Each dog's general conditions were observed regularly during the whole progress. The changes of superoxide dismutase, malondialdehyde, glutathione peroxidase in the serum were detected at regular time of the whole process, and the structure changes of each organ were observed at 24 weeks of treatment. RESULTS AND CONCLUSION:(1) Before treatment, the levels of superoxide dismutase and glutathione peroxidase in the aging control and treatment groups were lower than those in the young control group (P0.05). (2) For the aging treatment group at 24 weeks of treatment:the beagle fur became clearer and smoother, motility was strengthened, appetite became better;and the activity of superoxide dismutase in serum at 8 to 24 weeks of treatment increased significantly compared with before treatment (P<0.05), the activity of glutathione peroxidase significantly increased from 6 weeks (compared with treatment before, P<0.05), the malondialdehyde content decreased significantly from 4 weeks of treatment to the completion of the experiment (compared with before treatment, P<0.05). (3) After the experiment, the microscopic observation showed that compared with the aging control group, the tissues and organ structures of the aging treatment group were al clear, had no inflammatory infiltrates, no obvious necrosis and fibrosis lesions. These results were mainly consistent with the observations of young control group. The above results show that umbilical cord mesenchymal stem cel s combined with lycopene therapy on the natural aging beagles may enhance the activities of antioxidant enzymes, reduce malondialdehyde content, and their combination also can repair tissue structures and promote the functions, which has obvious anti-aging effects.
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In order to discover the mechanism of Xuebijing oral effervescent tablet (XBJOET) to treat infectious diseases, the effect of XBJOET on endotoxin induced rabbit fever and disseminated intravascular coagulation (DIC) was investigated. Auricle microcirculation in rabbit was detected by laser speckle blood perfusion imager system; coagulation function was measured by coagulation analyzer, fibrinolytic system was quantified by Elisa assay and micro thrombosis in tissues was observed with HE staining under light microscope. The results demonstrated that the body temperature of rabbit decreased significantly at 1-3 h after administration with 4.8, 2.4 and 1.2 g x kg(-1) XBJOET to endotoxin induced DIC rabbit model, the auricle microcirculation blood flow in model group (54.45 +/- 14.53) PU was lower than that in control group (77.18 +/- 12.32) PU. The auricle microcirculation blood flow increased markedly and there was significant difference between model group and 1.2 g x kg(-1) XBJOET group. There was significant difference between model group and control group in the content of PAI1 and FIB. The PAI1 levels in model and control groups were (30.48 +/- 2.46) ng x mL(-1) and (20.93 +/- 3.25) ng x mL(-1), respectively. The FIB levels in model and control group were (3.34 +/- 1.09) g x L(-1) and (4.84 +/- 1.10) g x L(-1), respectively. The content of PAI1 in rabbit plasma decreased notably, there were significant differences between model group and 4.8, 2.4 g x kg(-1) XBJOET groups. On the contrary the content of FIB increased. XBJOET possessed pharmacological activities of curing infectious fever and DIC, the mechanism of which is related to amelioration of microcirculation disturbance, inhibition of fibrinolytic system activation and coagulation and micro thrombosis elimination.
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This study is to investigate the treatment of Jin Chai antiviral capsule for influenza virus FM1/47 (H1N1) infection. The model of pneumonia was established by dropping influenza virus into the nose of normal mice, real-time PCR and Western blot technique were used to detect the virus load and the interferoninducible transmembrane protein3 (IFITM3) in lung of mice at the 1st day, 3rd day, 5th day and 7th day after affected. The results showed that Jin Chai antiviral capsule in large, middle, small dose groups can decrease virus load significantly at each time point, after being affected (P<0.05, P<0.01), Jin Chai antiviral capsule can increase the interferoninducible transmembrane protein3 in lung of mice, large dose groups are significantly higher in expression of IFITM3 compared with model group at each time point (P<0.05, P<0.01). Middle dose groups are significantly higher in expression of IFITM3 compared with model group at the 3th day and the 5th day (P<0.05), small dose groups are significantly higher in expression of IFITM3 compared with model group at the 3th day (P<0.05). It can be concluded that Jin Chai antiviral capsule exerts antiviral effects against influenzavirus by raised expression of IFITM3.
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Qingkailing injection, Shuanghuanglian injection, baicalin, chlorogenic acid as sample, guinea pig as control, to observe the specificity of allergic response to traditional Chinese medicine (TCM) injection in BN rats and establish a suitable animal model to evaluate applicability of allergic response in BN rats and guinea pigs induced by TCM. BN rats were sensitized by TCM injection, the symptoms, the rate and degree of allergic response were observed, the level of histamine in serum and tissues were determined by ELISA assay, the rate and degree of pathological changes in target organs were observed by HE staining under light microscope. There were significant symptoms of allergic response can be in BN rats, the level of histamine in serum, lung and trachea tissues increased significantly and there were significant pathological changes in lungs and tracheas. Meanwhile, the similar symptoms of allergic response can be induced by penicillin and trichosanthin. The rate and degree of allergic response, the rate and degree of pathological changes was higher in BN rats than in guinea pigs. Compared with guinea pig, BN rat is probably more suitable animal model in evaluating allergic response to injection of TCM.
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Animals , Male , Rats , Disease Models, Animal , Drug Hypersensitivity , Drugs, Chinese Herbal , Guinea Pigs , Hypersensitivity, Immediate , Injections , Medicine, Chinese Traditional , Rats, Inbred BNABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of Yinqiao detoxifcation oral liquid on activity of natural killer cells (NK) and serum content of TNF-alpha, TGF-beta1 of BALB/C nude mouse infected by influenza virus.</p><p><b>METHOD</b>To establish infected mice model by FM1 followed by intragastric administration of Yinqiao detoxifcation oral liquid for treatment. LDH method was used to observe NK cells. ELISA method was used to determine the levels of TNF-alpha, TGF-beta1, in serum on 1st, 3rd, 5th, 7th days after infection.</p><p><b>RESULT</b>Comparing to the normal group, the NK activity of the model group was significantly increased on 1 dpi (day post infection), and significantly decreased on 3, 5, 7 dpi. The NK activity of three dosage groups (5, 10, 20 g x kg(-1)) of Yinqiao detoxifcation oral liquid were respectively higher than that of the model on 3, 5, 7 dpi, especially with high dose (P < 0.01). The serum level of TNF-alpha and TGF-beta1 of model group is higher than that of normal group on 1, 3, 5, 7 d. Compared with model group, the serum level of Yinqiao detoxifcation oral liquid groups (5, 10, 20 g x kg(-1)) were decreased in different degree on every time point, especially the serum level of the higher dose of Yinqiao detoxifcation oral liquid decreased on 3 dpi (P < 0.05), Yinqiao detoxifcation oral liquid inhibit the serum level of TGF-beta1 in a dose-dependent manner.</p><p><b>CONCLUSION</b>Yinqiao detoxifcation oral liquid could enhance the activity of NK cell and decrease the serum level of TNF-alpha and TGF-beta1 of the mice infected by influenza virus.</p>
Subject(s)
Animals , Humans , Male , Mice , Administration, Oral , Disease Models, Animal , Drugs, Chinese Herbal , Influenza A virus , Allergy and Immunology , Physiology , Influenza, Human , Drug Therapy , Allergy and Immunology , Virology , Killer Cells, Natural , Allergy and Immunology , Mice, Inbred BALB C , Mice, Nude , Random Allocation , Transforming Growth Factor beta1 , Allergy and Immunology , Tumor Necrosis Factor-alpha , Allergy and ImmunologyABSTRACT
It is to investigate the effect of two kinds of Houttuynia Cordata Injection on preventing and treating H1N1 influenza virus infection in mice. Pneumonia model was set up by intranasal infection of the normal and immunocompromised mice with influenza virus FM1 and PR8. The two injections were administered before and after the administration of virus, separately, and the lung index was observed. The results showed that the two preparations have obvious therapeutic effect on normal mice infected with influenza virus FM1 and PR8. And to FM1, the new injection's effect is better at small dosage. The results also showed that the two preparations have obvious prophylactic effect on immunodepressed mice infected with influenza virus FM1 and PR8. And to PR8, the old injection's effect is better at small dosage. Houttuynia Cordata Injection can improve the mice pneumonia caused by influenza virus H1N1 and decrease the lung index markedly. It has a remarkable preventive and therapeutic effect on H1N1 influenza virus in mice.
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Objective To investigate the effect of the Gardenia extracts T9 on herpes simplex virus (HSV) viral protein 16(VP16) in HSV-1 infected mice's brains and its potential anti-viral mechanism. Methods HSV-1 infection was induced in BALB/c mice, the expression of HSV VP16 was detected by RTPCR on the 4th, 7th, 10th, 14th, and 21st day. Results The expression of VP16 mRNA decreased in both groups of large and low doses compared with virus group at the same time. Conclusion Gardenia extracts T9 can down-regulate the expression of VP16 mRNA in HSV infected mice's brains, and it may be one of its anti-HSV mechanisms.
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Aim To investigate the effect of simvastatin on the FKN expression in human umbilical vascular endothelial cells(HUVEC)up-regulated by interleukine-18(IL-18),and the effect on adhesion of FKN to monocyte THP-1.Methods FKN expression in HUVEC was determined by reverse transcription-polymerase chain reaction (RT-PCR), and the adhesion was checked by in vitro Flow chamber.Results Incubation of HUVECs with IL-18 upregulated FKN mRNA expression(P