Objective Using different concentrations of Coxsackievirus B3(CVB3)to infect young SD rats.To investigate the distribution of coxsackievirus B3(CVB3)in rat tissues and the immune response and inflammatory factors,to clarify the immunopathological mechanism of viral infection and provide an experimental basis for drug screening and efficacy evaluation.Methods Young SD rats(7 days old)were injected intraperitoneally with different doses of CVB3(TCID50=10-3.34/100 μL)and the proportions of lymphocyte subsets(CD4+,CD8+)in whole blood at days 4 and 8 were detected by flow cytometry.The CVB3 loads in the heart,liver,spleen,brain,kidney,and gastrointestinal tissues were detected by real-time fluorescence quantitative polymerase chain reaction,TNF-α and IFN-γ levels were detected by enzyme-linked immunosorbent assay,and histomorphologic changes were observed by hematoxylin and eosin staining.Results Different doses of CVB3 caused different degrees of diarrhea and decreased body mass in young rats.CVB3 was mainly distributed in the stomach,small intestine,large intestine,and stools,with the highest load in the large intestine and stools.The stock solution group(TCID50=10-3.34/100 μL)increased the proportion of CD8+T cells in the whole blood in young rats and decreased the CD4+/CD8+ratio(P＜0.05,P＜0.01).Compared with the nomal group high TNF-α and low IFN-γ expression were observed in the large intestine of young rats in the concentrate group(P＜0.05,P＜0.01),and submucosal edema and inflammatory cell infiltration were observed in the large intestine(cecum and rectum).There were no significant differences in the proportion of lymphocyte subsets,TNF-α and IFN-γ levels,and morphological changes in whole blood of young rats in the group 10-1,10-2,and 10-3(P＞0.05).Conclusions Different doses of CVB3 can induce infections in young SD rats.CVB3(TCID50=10-3.34/100 μL)causes pathological changes in the large intestine(cecum and rectum)in young rats,and high virus replication can increase levels of inflammatory factors and cause an imbalance of immune cells.CVB3 may have a unique pathogenic mechanism in young rats,providing a theoretical basis for developing evaluation strategies for drugs against CVB3 virus infections.

Objective Using different concentrations of Coxsackievirus B3(CVB3)to infect young SD rats.To investigate the distribution of coxsackievirus B3(CVB3)in rat tissues and the immune response and inflammatory factors,to clarify the immunopathological mechanism of viral infection and provide an experimental basis for drug screening and efficacy evaluation.Methods Young SD rats(7 days old)were injected intraperitoneally with different doses of CVB3(TCID50=10-3.34/100 μL)and the proportions of lymphocyte subsets(CD4+,CD8+)in whole blood at days 4 and 8 were detected by flow cytometry.The CVB3 loads in the heart,liver,spleen,brain,kidney,and gastrointestinal tissues were detected by real-time fluorescence quantitative polymerase chain reaction,TNF-α and IFN-γ levels were detected by enzyme-linked immunosorbent assay,and histomorphologic changes were observed by hematoxylin and eosin staining.Results Different doses of CVB3 caused different degrees of diarrhea and decreased body mass in young rats.CVB3 was mainly distributed in the stomach,small intestine,large intestine,and stools,with the highest load in the large intestine and stools.The stock solution group(TCID50=10-3.34/100 μL)increased the proportion of CD8+T cells in the whole blood in young rats and decreased the CD4+/CD8+ratio(P＜0.05,P＜0.01).Compared with the nomal group high TNF-α and low IFN-γ expression were observed in the large intestine of young rats in the concentrate group(P＜0.05,P＜0.01),and submucosal edema and inflammatory cell infiltration were observed in the large intestine(cecum and rectum).There were no significant differences in the proportion of lymphocyte subsets,TNF-α and IFN-γ levels,and morphological changes in whole blood of young rats in the group 10-1,10-2,and 10-3(P＞0.05).Conclusions Different doses of CVB3 can induce infections in young SD rats.CVB3(TCID50=10-3.34/100 μL)causes pathological changes in the large intestine(cecum and rectum)in young rats,and high virus replication can increase levels of inflammatory factors and cause an imbalance of immune cells.CVB3 may have a unique pathogenic mechanism in young rats,providing a theoretical basis for developing evaluation strategies for drugs against CVB3 virus infections.

Ferrets offer an advantage in nonclinical studies of anti-infective drugs because of their ability to be infected with and spread pathogenic microorganisms,especially viral strains,without the need for host adaptation.Additionally,the clinical symptoms exhibited by infected ferrets are very similar to those of humans.Although ferrets play a very important role in the research and development of antiviral drugs,the scope of their application remains limited.This may be related to the lack of corresponding national standards for laboratory animal feeding and application of ferrets as well as the lack of specific diagnostic and detection reagents.This paper summarizes the characteristics of ferrets as infectious disease models with a summary and analysis of the application direction of ferrets in anti-infective drug research.Our aim is to promote further standardization of the use of ferrets.

Objective This study established a model of invasive Aspergillus niger lung disease in immunosuppressed rats to provide theoretical support for the pharmacodynamic evaluation of anti-invasive pulmonary aspergillosis drugs and mechanism studies.Methods Sixty SD rats were randomly divided into a normal control group;cyclophosphamide control group,and cyclophosphamide+fungal infection low,medium,and high dose groups,with 12 animals in each group.General clinical observations were performed daily,and the serum levels of immunoglobulin(Ig)G and IgM and galactomannan(GM)were detected by ELISA on the 3rd and 7th days of modeling.Simultaneously,the ratio of CD4+and CD8+cells,content of white blood cells(WBCs)and neutrophils(Neu)in peripheral blood,the Aspergillus niger load in alveolar lavage,and morphological changes to rat lung tissue were observed.Results Rats in the cyclophosphamide control and cyclophosphamide+fungal infection groups showed reduced voluntary activity and erect hair after modeling,and rats in the cyclophosphamide+fungal infection group also had shortness of breath and audible wet rhonchi in the lungs.Compared with the normal control group,rats in the cyclophosphamide control group showed significant reductions in the levels of CD4+,WBC,Neu,IgG,and IgM in the blood,and their proportion of CD8+cells was significantly higher(P＜0.05,P＜0.01).Compared with the cyclophosphamide control group,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly reduced blood levels of IgG,IgM,and CD4+cells(P＜0.05,P＜0.01);while the cyclophosphamide+fungal infection low-,medium-,and high-dose groups had significantly reduced blood levels of WBC and Neu(P＜0.05,P＜0.01).Additionally,rats in the cyclophosphamide+fungal infection medium-and high-dose groups had significantly increased blood CD8+cells(P＜0.05,P＜0.01),Blood GM levels and the alveolar lavage Aspergillus niger load were significantly increased in rats in the cyclophosphamide+fungal infection low-,medium-,and high-dose groups compared with the cyclophosphamide control group(P＜0.05,P＜0.01).The lung tissues of the cyclophosphamide+fungal infection low-,medium-,and high-dose groups showed mycelial distribution and destruction of alveolar epithelium,increase of bronchial epithelial cup cells in the alveoli,and infiltration of inflammatory cells,and the degree of lesions was positively correlated with the modeling dose.Conclusions In this study,we used Aspergillus niger combined with cyclophosphamide immunosuppressant to construct a model of invasive Aspergillus niger lung disease.The duration of the disease was positively correlated with the concentration of bacterial fluid and modeling time,confirming that cellular immunity plays an important role in the pathogenesis of the disease.At the same time,Ig can also affect the development of invasive pulmonary aspergillosis,and it is speculated that the pathogenesis may be related to the level of Ig produced by humoral immunity.

ObjectiveTo explore the distribution mechanism of network modules in the combined treatment of liver cancer with Jiuwei Zhengxiao granules （JWZX） based on the analysis framework of module pharmacology. MethodThe cell experiment and the animal experiment were carried out to investigate the in vitro anti-liver cancer efficacy of JWZX of different concentrations and the effect on the survival time of H22 ascites tumor mice. By virtue of the analysis strategy of modular pharmacology，the distribution characteristics of nine Chinese drugs in the liver cancer disease network modules were investigated based on the constructed liver cancer disease network and module division by MCODE. In this study，the average degree （AD） of the nodes in the modules was used as an index to screen the main modules of the disease，and the intervention of the sovereign drugs，minister drugs，and assistant drugs on the main modules was explored. Finally，the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis was performed on the drug-acted modules by Metascape. ResultAs revealed by the cell experiment，JWZX could significantly inhibit the proliferation of H22 cells. The animal experiment demonstrated that the medium- and high-dose JWZX could significantly prolong the survival time of mice with H22 ascites tumor （P<0.05，P<0.01）. The distribution of targets of JWZX in the liver cancer disease network modules showed that JWZX interfered with tumor necrosis factor （TNF），epidermal growth factor receptor （EGFR），vascular endothelial growth factor A （VEGFA），transcription factor （JUN），tumor protein p53 （TP53），and other 26 targets and 8 modules. The sovereign drug Ginkgo Semen mainly intervened in modules 3 and 8，and the minister drugs such as Centipeda Herba jointly intervened in modules 1，3，5，8，10，and 12. Centipeda Herba and Phyllanthi Fructus intervened in module 7 and module 19 individually. Artemisiae Annuae Herba and other assistant drugs jointly intervened in modules 3，5，10，and 12. KEGG pathway enrichment analysis found that 135 pathways were enriched in 8 modules，and the pathway functions involved 12 categories including cancer，signal transduction，immune system，endocrine system，and amino acid metabolism. The functions of the four major modules involved cancer，signal transduction，and immune system. According to the results of literature verification，the key links of JWZX on the liver cancer disease network and the core mechanism were presumedly related to the inhibition of phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） and hypoxia-inducible factor-1 （HIF-1） signaling pathways，reduction of the immunosuppressive effect in the tumor microenvironment，and improvement of the anti-tumor immune response. ConclusionJWZX possesses pharmacological activity against liver cancer，and the therapeutic efficacy was achieved through the multiple targets，multiple modules，and multiple functions of drugs alone or in combination to intervene in the disease. The present study reduced the complexity of drug-disease target network analysis with module analysis strategy and explored the network module distribution mechanism of JWZX in the treatment of liver cancer，which provides a new idea for interpreting the complex mechanism of prescription compatibility.

OBJECTIVE: To detect variants of IVD gene among 4 neonates with suspected isovalerate acidemia in order to provide a guidance for clinical treatment. METHODS: 111 986 newborns and 7461 hospitalized children with suspected metabolic disorders were screened for acyl carnitine by tandem mass spectrometry. Those showing a significant increase in serum isovaleryl carnitine (C5) were analyzed for urinary organic acid and variants of the IVD gene. RESULTS: Four cases of isovalerate acidemia were detected, which included 2 asymptomatic newborns (0.018‰, 2/111 986) and 2 children suspected for metabolic genetic diseases (0.268‰, 2/7461). The formers had no obvious clinical symptoms. Analysis of acyl carnitine has suggested a significant increase in C5, and urinary organic acid analysis has shown an increase in isovaleryl glycine and 3-hydroxyisovalerate. Laboratory tests of the two hospitalized children revealed high blood ammonia, hyperglycemia, decreased red blood cells, white blood cells, platelets and metabolic acidosis. The main clinical manifestations have included sweaty foot-like odor, feeding difficulty, confusion, drowsiness, and coma. Eight variants (5 types) were detected, which included c.158G>A (p.Arg53His), c.214G>A (p.Asp72Asn), c.548C>T (p.Ala183Val), c.757A>G (p.Thr253Ala) and 1208A>G (p.Tyr403Cys). Among these, c.548C>T and c.757A>G were unreported previously. None of the variants was detected by next generation sequencing of 2095 healthy newborns, and all variants were predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics. CONCLUSION The incidence of isovalerate acidemia in Liuzhou area is quite high. Screening of metabolic genetic diseases is therefore recommended for newborns with abnormal metabolism. The discovery of novel variants has enriched the mutational spectrum of the IVD gene.
Infant, Newborn ; Child ; Humans ; Acidosis ; Carnitine ; Erythrocytes ; High-Throughput Nucleotide Sequencing

Objective:To discuss the clinical curative effect of the minimally invasive percutaneous suture technique of eight times for repairing closed injury extensor tendon zone I of finger.Methods:From February 2017 to January 2020, 12 patients (male 8, female 4) with mallet finger deformity were retrospectively studied, with an average age of 35 years (range, 18-50 years). And all the affected fingers were acute closed rupture of extensor tendon in zone I of single finger, 5 cases of the left finger and 7 cases of the right finger. There were 1 case of the thumb finger, 2 cases of the index finger, 3 cases of the middle finger, 4 cases of the ring finger and 2 cases of the little finger. 12 patients with fresh sputum mallet fingers were with 3-0 thread monofilament suture on extensor tendon zone I of finger in the minimally invasive percutaneous suture technique of eight times, and the distal end of the tendon was fixed to the base of the distal phalanx through the bone hole. Removal of the Kirschner wire 6-8 weeks, the brace was used to fix the affected finger in the dorsal extension. The flexion and extension of the affected finger was gradually strengthened. The function of the affected finger was evaluated according to the Crawford standard after operation and follow-up. The active flexion and extension range of motion of each joint of the affected finger and the contralateral healthy finger were measured, and the total action movement (TAM) of the finger were recorded. Finger function was evaluated according to TAM of the American Association of Hand Surgeons.Results:All operations were successfully completed, the operation time of the patients ranged from 18 to 25 min, with an average of 20.1±0.2 min. There was only a small amount of bleeding in the surgery. All 12 cases were followed up and the follow-up periods ranged from 6 to 14 months, with an average of 10.2±1.1 months. Mallet finger deformities were all corrected postoperatively; there were no knot exposure, skin necrosis and other complications. According to the Crawford standard, 9 cases were excellent, 2 cases were good, and 1 case was fair. The excellent and good rate was 91.7% (11/12). The mean active flexion of distal interphalangeal joints on the wounded finger and healthy finger were 82.11°±2.02° and 84.09°±2.01°, the mean active extension of distal interphalangeal joints on the wounded finger and healthy finger were -2.04°±3.01° and 0.02°±1.02°, there were significant differences between them ( t=2.447, 3.246; P=0.019, 0.004). The degrees of active joint activity of wounded finger were: 91.02°±4.01° of the metacar-pophalangeal joint, 94.04°±2.11° of the proximal interphalangeal joint, 83.01°±2.02° of the distal interphalangeal joint, and 265.05°±13.04° of total active activity; the degrees of active joint activity of healthy finger were: 93.01°±3.21° of the metacar-pophalangeal joint, 94.03°±3.07° of the proximal interphalangeal joint, 85.02°±2.01° of the distal interphalangeal joint, and 269.02°±12.10° of total active activity. The TAMs of the healthy side were 269.02°±12.10°, and the TAMs of the affected side were 265.05°±13.04°, there was no significant difference between them ( P>0.05). According to TAM system assessment criteria: excellent in 9 patients, good in 3 patients, and the excellent and good rate was 100% (12/12). Conclusion:The minimally invasive percutaneous suture technique of eight times can well repair closed injury extensor tendon zone I of finger, can have satisfactory treatment outcome in mallet finger with a simple procedure and good outcome. It is a simple, safe, effective method with minimal invasion.

OBJECTIVES: Cough variant asthma (CVA) is the main cause of obstinate cough. This study aimed to observe the therapeutic effect of Xiaochuan pill on CVA in a rat model, and to explore the mechanisms. METHODS: The rats were sensitized and challenged with 4% ovaibumin (OA) and 2% Al(OH) to establish the CVA models. They were treated with Xiaochuan pill (at the dose of 0.9, 1.8, 3.6 g/kg) or montelukast sodium once a day for 14 days. After 7 and 14 days of intervention, 5 and 10 rats were randomly selected from each group to collect bronchoalveolar lavage fluid (BALF), trachea, and lungs. The number of white blood cells (WBC) and eosinophils (EOS), and the levels of IL-1β, TNF- α, and IFN-γ in BALF were detected. Histopathological examination of lung tissue was performed to observe the histomorphological changes. The expressions of TLR4, MyD88, NF-κBp65, and p-p65 in lung tissue were detected by Western blotting. RESULTS: The numbers of WBC and EOS in BALF of CVA rats were significantly decreased by Xiaochuan pill (<0.05 or <0.01). The hyperplasia of tracheal, bronchial mucosa and the infiltration of inflammatory cells in lung were alleviated obviously. After 14 d of intervention, high dose of Xiaochuan pill significantly increased the level of IFN- γ (<0.01), reduced the levels of IL-1β (<0.05) and TNF-α (<0.05), and decreased the expressions of TLR4, MyD88, p65, and p-p65 (<0.05 or <0.01). CONCLUSIONS Xiaochuan pill exerts the significant therapeutic effect on obstinate cough in rats. The mechanism of action may be related to the regulation of TLR4-MyD88-NF-κBp65 signaling pathway as well as the inflammation and immune response.
Animals ; Cough ; Myeloid Differentiation Factor 88 ; NF-kappa B ; Rats ; Signal Transduction ; Toll-Like Receptor 4 ; Tumor Necrosis Factor-alpha

This study aimed to observe the therapeutic effect and mechanism of Jinshuibao tablet on acute renal injury induced by cisplatin. Acute renal injury models in SD rats were induced separately by single intraperitoneal injection of cisplatin(5 mg/kg)and intravenous injection for 5 consecutive days at a dosage of 2 mg/kg per day. The renal function and renal histopathological changes were observed in rat acute renal injury models after prevention and treatment with Jinshuibao tablet, respectively. The content of tumor necrosis factor(TNF-α)and reactive oxygen species(ROS), the activity of Caspase 3 and the expression of t-p38, p-p38, Bax and Bcl-2 in the kidneys were detected. The results showed that preventive and therapeutic administration of Jinshuibao tablets could both significantly inhibit the increase of the blood urea nitrogen(BUN)and creatinine(CRE), increase the creatinine clearance rate, reduce the contents of TNF-α and ROS, and decrease the activity of Caspase 3 in acute renal injury models induced by cisplatin. The renal histopathological results showed that Jinshuibao tablets could significantly reduce renal histopathology scores, ameliorate renal tubule degeneration and inflammatory infiltration. Western blot results showed that Jinshuibao tablets could significantly decrease the expression of t-p38 and p-p38, while increasing the Bcl-2/Bax ratio in the kidneys. These results suggested that preventive and therapeutic administration of Jinshuibao tablets could both improve renal function and pathological changes of renal tissue, which might be related to the inhibition of TNF-α and the ROS-p38 MAPK-Caspase3 pathway and thus inhibition of apoptosis.

Adipocytokines are polypeptides or proteins that are secreted by fat cells with a wide range of biological activities. Adiponectin is a fatty cytokine with insulin sensitization. It possesses the function of anti- diabetes, atherosclerosis and anti-inflammation. Adiponectin may participate in regulating the development of cognitive impairment, which is considered as a new regulatory factor for cognitive impairment.
Adiponectin ; Cognitive Dysfunction ; Diabetes Mellitus ; Humans ; Insulin ; Insulin Resistance