This study was aimed to observe the effect of Xiao-Chai-Hu Decoction (XCHD) with different doses of Bupleurum on body temperature and content of IL-1β, IL-6 and TNF-α in serum among fever rat models induced by lipopolysaccharide (LPS) in order to and investigate its action mechanism. A total of 50 rats were randomly divid-ed into 5 groups, including the normal control group, fever model group, XCHD group (high-dose), XCHD group (middle-dose), and XCHD group (low-dose). The fever rat model was established by intraperitoneal injection of LPS (20 μg/kg). At the same time, all rats were given corresponding XCHD or saline solution by gavage. The body tem-perature of rats (Ti) was measured every 30 min for 240 min after the injection of LPS. Change values of body tem-perature △T(△T=Ti-T0) were calculated. After the last measurement of body temperature, blood was taken for the de-tection of IL-1β, IL-6 and TNF-α level in serum with ELISA. The results showed that the body temperature of XCHD group (high-dose) was obviously decreased at the time point of 3 h, 3.5 h, 4 h (P< 0.01, or P< 0.05). This temperature reducing effect at the time point of 3.5 h and 4 h was better than the XCHD group (middle-dose) and XCHD group (low-dose, P< 0.01, or P< 0.05). Compared with the fever model group, all three XCHD groups can obviously reduce the IL-1β level (P< 0.05). Among them, the XCHD group (high-dose) had obvious effect (P<0.01). Compared with the fever model group, the XCHD group (high-dose) can obviously reduce contents of IL-6 and TNF-α in serum (P< 0.05). It was concluded that XCHD can reduce fever induced by LPS in rats and decrease contents of IL-1β, IL-6 and TNF-α in serum. This effect has a positive dose-effect relationship with the dose of Bu-pleurum in a certain range.

Chemical disinfectants are generally used for virus inactivation and environment disinfection in biosafety laboratory, and the efficacy and evaluation of the disinfection are critical to ensure the laboratory biosafety.However, there is a current lack of applied standard to evaluate the virucidal efficacy of chemical disinfectants in our country.In this paper, a European Union standard“Method and Requirements of Virucidal Quantitative Suspension Test Method for Chemical Disinfectants Used in Human Medicine” was analyzed and a standard transformation scheme has been proposed.It is suggested that the model viruses should be increased from 3 to 6, including the surrogate viruses to substitute highly pathogenic viruses, and that the method to remove the residual chemical disinfectant and the calculation of 95%confidence interval should be incorporated into the standard.The suggestion will improve the scientific and operational standards related to disinfection and sterilization in biosafety laboratory in China.

Objective To explore the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and the characteristics of human rotavirus(RV) G1P[8] infection to these cells,and establish a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8].Methods The primary small intestinal epithelial cells were obtained by collagenase Ⅺ and dispase I digestion from tree shrew.After purification and identification,the obtained primary small intestinal epithelial cells were infected with RV.Then,culture supernatants of infected cells were collected every 12 hours after infection.Viral titer and viral load were subsequently determined.Western blot and indirect immunofluorescence observation were used to detect the expression of RV protein VP6 in the primary cells.The infectivity of RV to the tree shrew primary cells was finally evaluated.Results After purification and identification of primary epithelial cells from the tree shrew,high purity above 90% primary tree shrew small intestinal epithelial cells was obtained.These primary small intestinal epithelial cells could be infected with RV virus by comparing the virus infectivity to primary renal cells,HCT116 cells and MA104 cells.The virus titer reached to 2.0×105TCID 50/mL at 72 h after infection.Using Western blot and indirect immunofluorescence observation,the specific viral protein of VP6 was determined to be expressed in the tree shrew primary small intestinal epithelial cells,and were located in the cytoplasm from days 1 to 5.Conclusions The separation,purification and cultivation methods of tree shrew primary small intestinal epithelial cells are successful,and the tree shrew model of RV-infected the tree shrew primary small intestinal epithelial cells is successfully established.

OBJECTIVETo screen the hepatocyte proteins that interact with hepatitis B virus X protein (HBx).

METHODSThe recombinant plasmid pSos-HBx was constructed by inserting Sos-HBx fragment into the bait vector, and after sequence verification the plasmid was transformed into competent yeast cells. The expression and self-activation of Sos-HBx protein was detected in the yeast cells. The hepatocyte proteins interacting with the bait protein was screened with CytoTrap yeast two-hybrid technique.

RESULTSThe reconstructed plasmid harboring HBx gene expressed Sos-HBx protein in the yeast cells without self-activation of the protein. CytoTrap yeast two-hybrid system identified 6 hepatocyte proteins that interacted with HBx, including fibronectin 1, translationally controlled tumor protein, IQ motif and WD repeats 1, follistatin, orosomucoid 1, and disulfide isomerase family A member 3.

CONCLUSIONSix HBx-binding hepatocyte proteins have been identified using the CytoTrap yeast two-hybrid system, which provides clues for further investigation of the role of HBx protein in hepatitis and liver cancer.

Genetic Vectors ; Hepatocytes ; metabolism ; Humans ; Plasmids ; Protein Interaction Domains and Motifs ; Proteins ; metabolism ; Trans-Activators ; metabolism ; Two-Hybrid System Techniques

OBJECTIVE： To study improvement effects of Fuling gancao decoction on functional dyspepsia （FD） model rats. METHODS： Sixty SD rats were randomly divided into normal group， model group， Fuling gancao decoction high-dose group （20 g/kg）， Fuling gancao decoction low-dose group （10 g/kg）， drug combination group （Fuling gancao decoction 10 g/kg+domperidone 0.004 g/kg）， domperidone group （0.004 g/kg）， with 10 rats in each group. Except for normal group， other groups were givenicy dilute hydrochloric acid intragastrically to establish FD model.  After modeling， normal group and model group were given constant volume of distilled water at room temperature intragastrically， and other groups were given relevant drug solution （1 mL/100 g） intragastrically， once a day， for consecutive 7 d. The amount of gastric residue was measured to evaluate the gastric emptying function. Immunohistochemistry SP method was used to detect the protein expression of AQP3， Ghrelin and Substance P in gastric mucosa tissue. The protein expression of Claudin-1， Occludin and VIP were detected by Western blot assay. RESULTS： Compared with normal group， the amount of gastric residue was increased significantly in model group （P＜0.01）； positive expression of AQP3 and Ghrelin protein in gastric mucosa tissue was decreased significantly， while the positive expression of Substance P protein tended to increase； the protein expression levels of AQP3， Ghrelin， Claudin-1 and Occludin were decreased significantly， while those of Substance P and VIP were increased significantly （P＜0.05 or P＜0.01）. Compared with model group， the amount of gastric residue was decreased significantly in administration groups （P＜0.05 or P＜0.01）； the positive expression of AQP3 and Ghrelin protein tended to increase， while less positive expression of Substance P was found； the protein expression levels of AQP3 and Occludin in gastric mucosa tissue were increased significantly in administration groups， while those of VIP were decreased significantly  （P＜0.05 or P＜0.01）； the protein expression levels of Ghrelin and Claudin-1 in gastric mucosa tissue were increased significantly in Fuling gancao decoction high-dose group， Fuling gancao decoction low-dose group and drug combination group， while those of Substance P were decreased significantly （P＜0.05）. CONCLUSIONS： Fuling gancao decoction may improve the symptom of fluid retention in FD model rats by up-regulating the protein expression of AQP3， Ghrelin， Claudin-1 and Occludin and down-regulating the protein expression of Substance P and VIP in gastric mucosa tissue of rats.

ObjectiveTo investigate the short-term efficacy of orthopedic elastic bandages on gait symmetry and walking ability in children with spastic hemiplegic cerebral palsy. MethodsFrom June, 2020 to June, 2023, 31 children with spastic hemiplegic cerebral palsy from Beijing Bo'ai Hospital were randomly divided into control group (n = 15) and experimental group (n = 16). Both groups received routine rehabilitation, while the control group received routine walking training, and the experimental group wore an orthopedic elastic bandage for walking training, for four weeks. The indexes of gait symmetry of foot deviation angle ratio (affected/healthy), step length ratio (affected/healthy), gait line ratio (affected/healthy) and standing stage ratio (affected percentage/healthy percentage) were calculated before and after training, and they were measured step width and the optional and maximum walking speed of 10-meter walk test (10MWT). ResultsOne case dropped off in the experimental group. After training, the foot deviation ratio, step length ratio, gait line ratio, and standing stage ratio improved in both groups (|t| > 2.434, P < 0.05), and they were better in the experimental group than in the control group (|t| > 2.230, P < 0.05); while the optional and maximum walking speed of 10MWT improved in both groups (|t| > 9.186, P < 0.001), and they were better in the experimental group than in the control group (|t| > 2.278, P < 0.05). ConclusionWearing orthopedic elastic bandages during rehabilitation can promote the gait symmetry and walking ability of children with spastic hemiplegic cerebral palsy.