Objective Discuss the interference of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin .Methods Collected 60 examples ,include total bilirubin concentration 20 examples less than 20 μmol/L ,20 examples between 150-220 μmol/L and 20 examples between 350-410 μmol/L ,add an equal volume of various concentrations of cefotiam in each case ,formulated into cefotiam final concentrations of 300 ,150 ,75 mg/L of serum samples as the test group ,add an equal volume of water in each serum samples as the control group ,determine all the samples total bilirubin concentration respectively by vanadate oxidation method and dry chemical method ,compared the interference of cefotiam on determined total bilirubin by two method ,analyze the data by SPSS13 .0 .Results Determined total bilirubin by dry chemical method ,the test group higher than the control group ,the difference was statistically significant(P<0 .05) ,at the same total bilirubin levels ,with cefotiam concentrations decreased ,increased rate of total bilirubin concentration were decreased in the experimental group .Determined total bilirubin by vanadate oxidation method ,when the total bilirubin concentration between 150 -220 μmol/L ,the test group was higher than the control group ,the difference was statistically significant(P<0 .05) .Conclusion Interference of injection cefotiam on determined to‐tal bilirubin by dry chemical method is strong ,and with the drug concentration increased ,effect is more obvious ,but determination of total bilirubin by vanadate oxidation method has almost no effect .

Objective Using middle cerebral artery occlusion (MCAO) model to observe protective effect of effective components of bezoar on brain damage.To discuss the anti-cerebral ischemia mechanism and compare the efficacy of effective components of bezoar from the endoplasmic reticulum stress intervention angle.Methods Rats were stratified randomly divided into sham group,model group,Qingkailing group (positive drug,3 mL/kg),taurine group,ursodeoxycholic acid (UDCA,78 mg/kg) group,taurine-conjugated ursodeoxycholic acid (TUDCA,100 mg/kg) group.Through establish MCAO model in rats,observed the scores of the neurologic impairment,measured infarct volume by TTC.Immunohistochemistry and Western blotting were Used to detect the content of P-PERK,P-EIF2α,and ATF4.Results Compared with sham group,neurologic impairment scores of model group significantly reduced (P ＜ 0.01).Compared with model group,Qingkailing group,UDCA group,and TUDCA group significantly improved neurological function in rats (P ＜ 0.05,0.01).Compared model group,all the treatment groups could significantly reduce the volume of cerebral infarction (P ＜ 0.01).Compared with sham group,expression of P-PERK,P-EIF2α,and ATF4 was significantly increased (P ＜ 0.01).Compared with model group,all the treatment groups reduced the expression of P-PERK,P-EIF2α,and ATF4 in varying degrees,effect of Qingkailing and TUDCA were more obvious.Conclusion The effective components ofbezoar alleviate cerebral ischemia reperfusion injury in rats by inhibiting endoplasmic reticulum stress,the effect of TUDCA is better than that of taurine and UDCA.

Objective: To investigate the effects of EI injection on learning and memory ability and brain energy of two-way Meynert basal injection of Ibotenic acid (IBO) dementia model rats. Methods: A rat model of dementia wasestablished by bilateral meynert basal injection of IBO. After 8 weeks of EI injection, Morris water maze was used todetect the learning and memory ability of rats. Congo red staining was used to observe the deposition of Aβ plaque inhippocampal CA1 and cortical areas of rats. The changes of ATP, ADP and AMP in brain tissue of each group weredetermined by HPLC. The content of insulin in rat brain tissue was detected by ELISA kit. The expression of key proteinin PI3K/AKT signaling pathway was detected by Western blot. Results: Compared with the sham operation group, theescape latency of the model group was significantly prolonged, the number of entering the platform, the time andpercentage of crossing the platform quadrant decreased significantly (P ＜ 0.05); Aβ plaque deposition was observed inthe hippocampus and cortex; ATP/AMP ratio and insulin content were significantly decreased (P ＜ 0.05); brain tissue PI3 K and AKT protein were low expression (P＞ 0.05) . After intervention with EI injection, the escape latency of themodel rats was significantly shortened, the number of entering the platform and the time of crossing the platform quadrantincreased significantly (P ＜ 0.05); the hippocampus and cortex red staining was alleviated; the brain tissue ATP/AMPratio and insulin content increased significantly (P ＜ 0.05) . Conclusion: EI injection can improve the learning andmemory function of IBO-induced dementia model rats, which is related to the improvement of brain energy.

Objective: To investigate the effects of Kaixin Powder on learning and memory, ATP/AMP ratio, GABA andiNOS levels in rats with multiple infarct dementia. Methods: The rat model of multi-infarct dementia was established bymicro-thromboembolism. Morris water maze and opening experiment were used to evaluate learning and memoryfunction. The pathological changes of hippocampal CA1 area were evaluated by HE staining. The contents of ATP andAMP in brain tissue were determined by HPLC. The content of γ-aminobutyric acid and iNOS in peripheral blood weredetected by ELISA kit. Results: Compared with the rats of model group, rats of Kaixin Powder group can significantlyshorten the escape latency, increase the number of crossing platforms, Results: Compared with the model group, increasethe standing times in the opening experiment, prolong the exercise time, shorten the resting time, improve the nerve celldamage in the hippocampal CA1 area, and significantly increase the ratio of ATP/AMP of brain tissue, decreased brainGABA content and serum iNOS content (P ＜ 0.05) . Conclusion: Kaixin Powder has the effect of improving learning andmemory function and abnormal motor behavior in rats with multiple infarct dementia. The mechanism is related toreducing GABA and i NOS content in brain tissue, increasing ATP/AMP ratio in brain tissue and improving energysupply in brain tissue.

Integrins are transmembrane glycoproteins, closely related to many physiological and pathological processes. In order to explore its role in silkworm, by PCR and Rapid-amplification of cDNA ends (RACE) technology, the full-length cDNA of Bmintegrin β1 in silkworm was acquired. The domain was predicted by domain prediction website. Phylogenetic tree was constructed to analyze its evolutionary relationship. By prokaryotic expression system, protein purification method and immunizing mouse, the antibody against Bmintegrin β1 recombinant protein was obtained. The spatial-temporal expression profile of Bmintegrin β1 was investigated by semi quantitative PCR and Western blotting. Then we identified all 3 different spliceosomes, and they shared a common open reading frame of 2 502 bp, encoding 833 amino acids. Bmintegrin β1 contained all the classic domains of the integrin family, such as Integrin-B-tail, transmembrane domain etc. Phylogenetic analysis indicated that Bmintegrin β1 was close to the homologous proteins from Heliothis assulta and Danaus plexippus. In order to understand the function of Bmintegrin β1 further, we generated the antibody. In addition, Western blotting demonstrated that the antibody recognized the Bmintegrin β1 recombinant protein. Then, semi quantitative PCR and Western blotting results showed that Bmintegrin β1 was widely expressed in most of tissues, among of them, it's exhibited the highest expression level in hemacyte. Overall, this study provides a foundation for the study of silkworm integrin family.

Scavenger receptor class B is involved in various indispensable physiological processes, like the formation and inhibition of atherosclerosis or other cardiovascular diseases, innate immune defense and the removal of apoptotic cells. Here, we cloned BmSCRB8, a member of scavenger receptor class B in silkworm. We obtained the full-length cDNA sequence of BmSCRB8 by rapid amplification of cDNA ends (RACE), including 2 668 bp. The ORF of BmSCRB8 is 1 704 bp, encoding 567 amino acids. Online software prediction indicated that the molecular weight of BmSCRB8 is 63.87 kDa and the isoelectric point (pI) is 6.06. The space-time expression profile of BmSCRB8 was detected by reverse transcription PCR (RT-PCR), which implicated that BmSCRB8 is extensively expressed in each tissue and at each stage of blood. In addition, BmSCRB8 is highest expressed in fat body of silkworm, and is highly expressed in metamorphosis periods. Anti-BmSCRB8 polyclonal antibody was generated through prokaryotic expression, protein purification and mice immunization. Simultaneously, we constructed BmSCRB8 eukaryotic vector and then transfected embryonic cell line of silkworm. Immunofluorescence and overexpression showed that BmSCRB8 expressed specifically in membrane. Western blotting demonstrated that BmSCRB8 protein can be specifically recognized by anti-serum generated after mice immunization.

Integrins are cell adhesion receptors, which consists of several transmembrane glycoproteins. They are widely distributed on the cell surface and involved in signal transduction pathways. As a heterodimer, each integrin is composed of one α subunit and one β subunit. Integrins are mainly expressed on lepidopteran hemocytes and involved in cell immune response. The full-length cDNA sequence of BmIntegrin β2 was obtained by PCR and RACE, including 2 434 bp. BmIntegrin β2 was predicted to be a transmembrane protein. The BmIntegrin β2 expression profile was detected by qRT-PCR at L4D3 or L5D3 larval stage, and it was highly expressed in hemocyte and hematopoietic organ. Anti-BmIntegrin β2 polyclonal antibody was generated following prokaryotic expression, protein purification and animal immunization, which is highly specific and effective for recognizing BmIntegrin β2 protein through Western blotting. The results of plasmatocytes adhesion experiment showed that BmIntegrin β2 plays an important role on the adhesion and spreading of plasmatocytes to foreign surfaces. This study provides a foundation for further research of the biological function of BmIntegrin β2 gene.

Endangered animal medicine is an important part of traditional Chinese medicine， which is distinctive in the treatment of diseases. At present， the rare and endangered medicinal materials such as tiger bone， rhinoceros horn， pangolin， antelope horn， bear bile are listed as national key protected animals， so their clinical application is limited， the current solution is mainly based on the ideas and methods of similar pharmacological effects， close genetic relationships， artificial breeding， and artificial synthesis to find and develop alternatives for endangered animal medicinal materials. Although artificially cultured bear bile and musk， and artificially synthesized tiger bone， bezoar and musk can solve the shortage of endangered animal medicines to a certain extent， there are still some problems such as difficult breakthroughs in breeding technology and incomplete recognition in the substitute industry. According to this， based on summarizing the existing substitutes for endangered animal medicines， our group proposed the concept of homology， homogeneity and equivalent of substitutes， and constructed a new idea to develop and evaluate substitutes by combining frontier biotechnology with multi-omics detection， so as to provide some support for protecting rare and endangered animals and solving the shortage of endangered animal medicines.