Objective:To investigate the effects of sorafenib on hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) levels and recurrence in patients with intrahepatic cholangiocarcinoma with microvascular invasion.Methods:Ninety-two patients with intrahepatic cholangiocarcinoma who received treatment in Yiwu Central Hospital between November 2013 and November 2019 were included in this study. They were randomly assigned to undergo either conventional basic treatment (control group, n = 46) or conventional basic treatment and sorafenib treatment (study group, n = 46). Clinical efficacy, the incidence of adverse reactions and recurrence rate were compared between the two groups. Before and after treatment, HIF-1, alpha fetoprotein (AFP) and VEGF levels were also compared between the two groups. Results:After treatment, total effective rate in the study group was significantly higher than that in the control group [63.04% (29 /46) vs. 28.26% (13/46), χ2 = 11.215, P < 0.05]. After treatment, HIF-1, AFP and VEGF levels in each group were significantly lower than those before treatment (all P > 0.05). After treatment, HIF-1 [(165.23 ± 39.67) pg/mL], AFP [(109.16 ± 67.31) ng/mL] and VEGF [(297.28 ± 42.41) pg/mL] levels in the study group were significantly lower than those in the control group [(205.56 ± 40.23) pg/mL, (235.17 ± 106.41) ng/mL, (365.16 ± 40.91) pg/mL, t = 4.841, 6.788, 7.813, all P < 0.05]. There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Six-month follow-up revealed that the incidence of recurrence in the study group was significantly lower than that in the control group ( χ2 = 4.792, P < 0.05). Conclusion:Sorafenib can reduce the HIF-1, AFP and VEGF levels in patients with intrahepatic cholangiocarcinoma with microvascular invasion, improve the clinical efficacy, decrease the incidence of recurrence, but cannot increase the incidence of adverse reactions.

Objective To compare coagulation data measured with domestic produced and imported coagulation testing solutions on SD rat and human by testing prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time(TT), fibrinogen(FIB).Methods Blood samples were obtained from SPF SD rat and human .Domestic produced and imported coagulation testing solutions were applied to test PT , APTT, TT, and FIB.Results Compared to rat data measured with imported coagulation testing solution , data measured with domestic produced coagulation testing solution of PT, APTT, FIB was significantly higher (P<0.05), while, data of TT was statistically lower(P<0.05), and there was no obvious difference in human blood coagulation .Conclusion The data measured with different coagulation testing solution varies on SD rat , so the laboratories are required to establish reference data according to different products .

This paper was mainly on pesticide degrading microorganisms. The major pesticide degrading bacteria were Pseudomonas, Bacillus, Flavobacterium, Alcaligenes, Arthrobacter, etc. The major pesticide degrading fungi were Aspergillus,Pinicielium, Rhizopus, Trichoderma, Fusarium, etc. The major pesticide degrading actinomycetes were Nocardia, Streptomyces, etc.

OBJECTIVE:To optimize the extraction technology of Bushen huoxue capsules. METHODS:Central composite de-sign-response surface method was used to optimize the extraction technology with the amount of water and boiling time as main fac-tors using normalized value of the contents of icariin and salvianolic acid B,the yield of dry extract as index. Validation test was al-so conducted. RESULTS:The optimal extraction technology was as follows as 15-fold water,reflux extracting for 75 min,extract-ing for 2 times. The deviation of measured value and predicted value was 0.000 3% in validation test. CONCLUSIONS:The cen-tral composite design-response surface method can be used to optimize the extraction technology of Bushen huoxue capsules with good prediction,and the technology is simple and stable.

Objective To study the influence of different routes of keyhole limpet hemocyanin ( KLH) immuniza-tion on the T-cell-dependent antibody response in mice.Methods SPF Kunming mice were divided into four groups: the intravenous injection group, subcutaneous injection group, intraperitoneal injection group and control group.Each mouse was injected 200 μg KLH intravenously, subcutaneously or intraperitoneally daily for consecutive 10 days, respectively. Mice in the control group were given solvent injection only.Serum concentration of IgG stimulated by KLH antigen was measured 7 days after the last dosing.Spleen was isolated to calculate the organ coefficient and examined by pathology u-sing hematoxylin and eosin staining.Results Intravenously, subcutaneously and intraperitoneally administered KLH stimu-lated the generation of secondary lymphoid follicles and germinal center to varying degrees, B cell apoptosis, increased a-mount of cells in the marginal zone and other pathological changes were observed in the spleen.Intravenous and intraperito-neal administration of KLH led to more pronounced pathological changes compared with that in the subcutaneous injection group.All of the three administration routes of KLH induced generation of IgG antibody, significantly higher than that in the control group (P<0.05).Intravenous injection of KLH generated the highest concentration of IgG and organ coefficient among the three administration routes ( P<0.05) .Conclusions Different immunization routes do affect the production of IgG antibody, organ coefficient and pathological changes in the spleen, and these differences should be taken into consider-ation when analyzing the T cell dependent antibody response in mice.

Objective To track the effects of hyperbaric oxygen (HBO) therapy on neonatal rats with hypoxic-ischernic brain damage (HIBD).MethodsA total of 126 healthy,seven-day-old SD rats were used to model the HIBD and then randomly divided into seven groups of 18:a sham group,a hypoxic-ischemic (HI) group,a 6 h HBO group,a 24 h HBO group,a 48 h HBO group,a 72 h HBO group and a 1 week HBO group.One hour of HBO treatment at 2 atmospheres absolute pressure was administered once daily for a week to the rats in the latter 5groups,starting at 6,24,48,72 hours and 1 week post the HIBD modeling,while no HBO was administered to the sham and HI groups.The effects were assessed in terms of histological changes ( the neuron density and the percentage of neuron apoptosis in the CA1 region of the hippocampus) and nitric oxide (NO),malondialdehyde (MDA) and superoxygen dismustase (SOD) concentrations after 15 days.ResultsIn the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups,average neuron density in the CA1 region was significantly higher than in the HI group.But in the 1 week HBO group the average density was not significantly different than in the HI group.In the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups the average percentage of neuron cell apoptosis in the CA1 area of the hippocampus was significantly lower than in the HI group,but the 1 week HBO group again showed no significant difference.There were significant differences in average NO,MDA or SOD levels among the 6 h HBO,24 h HBO,48 h HBO,72 h HBO and HI groups,but the 1 week HBO group showed no significantly higher average levels than the HI group.ConclusionsThe optimal therapeutic window for using HBO to protect against HIBD is within the first week.The best effect can be obtained in the first 6 hours,but after 1 week HBO no longer has a significant effect.The earlier the HBO is administered,the better the effect obtained.

We amplified genes encoding UDP-glucose dehydrogenase, ecohasB from Escherichia coli and spyhasB from Streptococcus pyogenes. Both ecohasB and spyhasB were inserted into T7 expression vector pRX2 to construct recombinant plasmids pRXEB and pRXSB, and to express in E. coli BL21(DE3). After nickel column purification of UDP-glucose dehydrogenases, the enzymes were characterized. The optimum reaction condition of spyHasB was at 30 °C and pH 10. The specific activity reached 12.2 U/mg under optimum condition. The optimum reaction condition of ecoHasB was at 30 °C and pH 9. Its specific activity reached 5.55 U/mg under optimum condition. The pmuhasA gene encoding hyaluronic acid synthase was amplified from Pasteurella multocida and ligated with ecohasB and spyhasB to construct the coexpression vectors pBPAEB and pBPASB, respectively. The co-expression vectors were transformed into E. coli BW25113. Hyaluronic acid (HA) was produced by biotransformation and the conditions were optimized. When recombinant strains were used to produce hyaluronic acid, the higher the activity of UDP-glucose dehydrogenase was, the better its stability was, and the higher the HA production could reach. Under the optimal conditions, the yields of HA produced by pBPAEB/BW25113 and pBPASB/BW25113 in shake flasks were 1.52 and 1.70 g/L, respectively, and the production increased more than 2-3 folds as previously reported.
Biotransformation ; Escherichia coli ; enzymology ; Genetic Vectors ; Glucuronosyltransferase ; genetics ; Hyaluronan Synthases ; Hyaluronic Acid ; metabolism ; Pasteurella multocida ; enzymology ; Streptococcus pyogenes ; enzymology ; Uridine Diphosphate Glucose Dehydrogenase ; metabolism

Equipment Design ; Facial Injuries ; therapy ; First Aid ; instrumentation

OBJECTIVETo verify the pharmacological hypothesis of prescriptions by studying the targeted distribution of major components in stewed rhubarb in the rat model with acute pancreatitis (AP).

METHODNormal SD rats (control group, n = 5) and the AP model induced with intraperitoneal cerulein (model group, n = 5) were taken as the experimental objects. Rats of the two groups were orally administered with stewed rhubarb granules (20 g x kg(-1)). Their heart, liver, spleen, lung, kidney and pancreas were collected two hours after the administration. Such constituents as emodin, chrysophanol, physcion, rhein and aloe-emodin and their concentrations in each tissue homogenate were detected by high performance liquid chromatography-mass-mass.

RESULTAloe-emodin and physcion in stewed rhubarb whose concentrations in liver and kidney of normal rats were higher than that in pancreatic tissues, while the distribution spectrums and concentrations of the remaining components in pancreatic tissues had no significant difference with that of other organs. The concentrations of emodin, aloe-emodin, rhein and chrysophanol in stewed rhubarb in pancreatic tissues of the AP model group were higher than that in other tissues and organs, while their concentrations in pancreatic, renal and splenic tissues were notably higher than that in the normal group.

CONCLUSIONIn the conditions of AP, effective components in stewed rhubarb show a targeted distribution feature in pancreas, which provides experimental basis for the pharmacological hypothesis of prescriptions.

Acute Disease ; Animals ; Anthraquinones ; pharmacokinetics ; pharmacology ; therapeutic use ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Organ Specificity ; Pancreatitis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).