AIM: To observe the effects of high-dose hepatitis B surface antigen (HBsAg) vaccine on cellular immune response in BALB/C mice. METHODS: The mice were immunilized separately with low-dose and high-dose HBsAg vaccine by intramuscular injection two times. The specific proliferative activities of T lymphocytes were measured by [ 3H]-TdR incorporation assay. IL-2 as well as IFN-? levels in the culture supernatant of T cells and anti-HBs IgG2a lever in sera were detected by enzyme-linked immunoabsorbent assay. RESULTS: After first vaccination with high-dose HBsAg, the proliferative activities of T cells in the experimental group were significantly stronger, both levels of IL-2 and IFN-? were markedly higher than that in the control group and the percentage of mice to produce serum anti-HBs IgG2a was significantly higher compared to that of mice immunilized by low-dose HBsAg. All data in experimental groups were further increased after second dose of vaccine. CONCLUSION: Vaccination of mice with high-dose HBsAg can induce cellular immune responses tended to Th1(T helper 1 subset) response.

We have constructed an immunotoxin(Ng76-TCS),which was composed of a monoclonalantibody directed against human melanoma and trichosanthin(TCS)——a single chain ribosomeinactivating protein.The cultured human melanoma cells(M21)were inhibited effectively byNg 76-TCS.The cytotoxicity of Ng76-TCS to M21 cells was 2,000-fold higher than that of free TCS and Ng76 mixture.A conjugate,which was prepared with normal mice immunoglobulinand TCS(NIgG-TCS),was 160-fold less cytotoxic to M21 cells.Meanwhile Ng76-TCS was125-fold less cytotoxic to nontarget cells Hela.These results showed that the immunotoxinNg76-TCS was a potent and specific anti-human melanoma agent.

Objective To establish a three-dimensional (3D) model of the diaphragm type of Budd-Chiari syndrome (BCS) with perforated membrane,to analyze changes of hemodynamic parameters pre and post percutaneous transluminal angioplasty (PTA) and to analyze any possibly related mechanical factors for postoperative recurrence in BCS.Methods The data on enhanced MRI from patients suffering from BCS with perforated diaphragm of inferior vena cava were reconstructed into a 3D model Computational fluid dynamics (CFD) were applied for numerical simulation based on the model,and the results were expressed as cloud images.Results The total number of model units of the finite element model for preoperative inferior vena cava stenosis was 16 422,and the total number of nodes was 48 170.The total number of model units for postoperative inferior vena cava was 16 539,and the total number of nodes was 51 339.Hemodynamic patterns of the lesion areas were effectively reflected in the 3D flow dynamic model for BCS.By comparing the hemodynamic parameters before and after interventional therapy,the data indicated that there was a gradual decline in wall pressure from the telecentric side.The largest blood flow velocity as well as wall shear stress were observed in the stenotic area of inferior vena cava.Both a large postoperative vascular central velocity and a low increase in local pressure predicted occurrence of vascular restenosis.Conclusions The establishment of a 3D fluid model of inferior vena cava revealed that there were mechanical changes in the location of the lesion.Variations in blood flow patterns exert a far-reaching influence on distribution of hemodynamic parameters,including local blood flow velocity,vascular wall pressure and shear stress.All these might be related to recurrence of BCS after interventional therapy.

Objective To study the characteristics of event-related potentials P300 in patients with anxiety disorder(AD). Methods P300 tests were carried out in 30 patients with AD and 30 healthy adult controls. ResultsPatients with AD had significantly delayed P3 latency ([326?16] ms vs [339?19]ms, P

Objective To explore the clinical application of 3D-printing model in guiding interventional management of Budd-Chiari syndrome (BCS) and in teaching practice.Methods A patient with typical BCS of inferior vena cava type was selected.By using MR enhanced scanning,the original MRA data of DICOM format were extracted,and then the digital data were extracted and reconstructed to obtain 3D BCS model by Simpleware software.The 3D BCS entity model,using 1 ∶ 1 ratio,was printed out by a 3D printer.An experienced chief physician made a simulated interventional manipulation on this 3D BCS entity model.Results The BCS 3D model was successfully printed.Simulated operation could be easily performed on the 3D-printing model,in this way the chief physician could make a demonstration of interventional procedure of BCS to the junior doctors and medical students.Interventional therapeutic manipulation for BCS could be well demonstrated on the 3D-printing model of BCS,which was very helpful in guiding teaching practice and in promoting the communication between doctors and patients.Conclusion The BCS 3D-printing model can truly reflect the spatial architecture features of the inferior vena cava and the hepatic veins,which are very valuable for the making of surgical plan,for the demonstration of simulation operation,and for teaching practice.Moreover,3D-printing model can stereoscopically display the location and morphology of the lesion,which can improve patient's understanding of the disease,thus,the communication between doctors and patients can be strengthened.

Abstract: Objective　To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods　According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results　The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion　The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.

Growth year is one of the important factors for the quality of Polygala tenufolia. In this study, primary metabolites and secondary metabolites were compared in 1, 2 and 3 years old P. tenufolia cultivated in Shaanxi Heyang. The samples were subjected to ultra-high performance liquid chromatography (UPLC)-quadrupole time-of-flight mass spectrometry (Q-TOF MS) and nuclear magnetic resonance (NMR) analysis, and the obtained data were analyzed using principal component analysis (PCA) and other statistical analysis methods. In addition, content and correlation of different metabolites were also calculated. The results showed no significance between main component contents in 2 year-old and 3 year-old P. Tenufolia, but 1 year-old was statistically different. The contents of primary metabolites, such as fructose, sucrose, and choline increased as time goes on, while glycine and raffinose decreased. The contents of secondary metabolites, such as onjisaponin Fg, polygalasaponin XXVIII, polygalasaponin XXXII increased, while polygalaxanthone III and parts of oligosaccharide multi-ester including tenuifoliose A, tenuifoliose C, tenuifoliose C2 and tenuifoliose H decreased with the extension of the growth years. Growth years has important impact on the quality of P. tenuifolia and the existing growing years of commodity P. tenuifolia have its scientific evidence. This study supplied a new method for the quality evaluation of Chinese medicinal materials.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Metabolomics ; Oligosaccharides ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Quality Control

OBJECTIVETo evaluate the benefits of biofeedback therapy in children with functional constipation caused by pelvic floor dysfunction (PFD).

METHODAnorectal manometry (PC Polygraf HR, Medtronic) was performed in children with functional constipation according to the diagnostic criteria of Rome III. Among them, 47 cases with PFD were trained with biofeedback therapy (PC Polygraf HR, Medtronic) according to the degree of cooperation. They received the treatment twice every week, and those in whom the therapy was performed at least three times were enrolled to evaluate the efficacy in this study.

RESULTA total of 27 cases (20 male and 7 female) in whom the procedure was performed at least three times were eligible for inclusion into this study. The mean age of them was (6.7 + or - 2.2) years (range 4 to 12 years), and the mean duration of symptoms was (3.0 + or - 2.3) years with a range of 6 months to 8 years. Among them, 16 cases received the biofeedback training more than 5 times, while 8 cases more than 7 times. The rectal maximum contraction pressure during defecation was increased significantly with the number of biofeedback training before treatment and after 3, 5, and 7 times of treatment was (36.2 + or - 10.4), (45.1 + or - 9.5), (47.6 + or - 9.1), and (47.8 + or - 8.8) mm Hg (1 mm Hg = 0.133 kPa) respectively (P < 0.01). The potential of electromyography of external anal sphincter decreased progressively, but had not reached significance. Follow-up continued for 3 to 12 months by telephone, the clinical response to biofeedback treatment was evaluated as excellent (complete resolution of constipation), good (improvement of constipation), partial or poor (no improvement of constipation). The response was excellent in 13 cases (48.1%), good in 8 (29.6%), partial in 3 (11.1%), and 2 (7.5%) cases had no improvement, and 1 case was lost to follow-up, and the rate of success was 88.9% (24/27).

CONCLUSIONBiofeedback therapy is a safe and effective treatment option for functional constipation due to PFD in children probably by increasing the rectal maximum contraction pressure during defecation.

Biofeedback, Psychology ; Child ; Child, Preschool ; Constipation ; therapy ; Female ; Humans ; Male ; Treatment Outcome

OBJECTIVETo study the antiproliferation effect on HepG2 cells and its underlying mechanism of the active chemical composition of the Viburnum Odoratissimum.

METHODS3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay and trypan blue dye exclusion assay were used to assess the effect of vibsane-type diterpenoids on the proliferation of various tumor cells. Alterations in cell cycle and apoptosis were determined by flowcytometry. The enzymatic activity of caspase-3/7 was measured by Apo-ONE homogeneous Caspase-3/7 Assay kit.

RESULTSCompound 1 #, a vibsane-type diterpenoid, was found to significantly inhibit the growth of HepG2 cells by anticancer proliferation activity screening. It was demonstrated that the modified groups on side chain coupled to C11 site affected the cell growth-inhibition activity of compounds by structure-activity analysis. In addition, HepG2 cell line was most sensitive to compound 1 #, which induced growth arrest of HepG2 cells in a dose- and time-dependent manner. Study on the mechanisms underlying these effects indicated that compound 1 # induced significant G0/G1 phase arrest of HepG2 cells in a time- and concentration-dependent manner. Meanwhile, It was found that higher concentrations of compound (5-10 micromol/L) caused evident increase in the unmber of apoptotic cells and dose-dependent activation of caspase-3/7.

CONCLUSIONVibsane-type diterpenoids could significantly inhibit the growth of HCC HepG2 cells. Induction of cell cycle arrest and apoptosis may play important roles in their anticancer effects.

Apoptosis ; drug effects ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Hep G2 Cells ; Humans ; Viburnum ; chemistry

Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality.
Angelica sinensis ; chemistry ; Animals ; Biological Assay ; methods ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Count ; Erythrocytes ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Plant Roots ; chemistry