Cloning and expression analysis of U6 promoters in Panax quinquefolius / 中国中药杂志

The U6 promoter is an important element driving sgRNA transcription in the CRISPR/Cas9 system. Seven PqU6 promo-ter sequences were cloned from the gDNA of Panax quinquefolium, and the transcriptional activation ability of the seven promoters was studied. In this study, seven PqU6 promoter sequences with a length of about 1 300 bp were cloned from the adventitious roots of P. quinquefolium cultivated for 5 weeks. Bioinformatics tools were used to analyze the sequence characteristics of PqU6 promoters, and the fusion expression vectors of GUS gene driven by PqU6-P were constructed. Tobacco leaves were transformed by Agrobacterium tumefaciens-mediated method for activity detection. The seven PqU6 promoters were truncated from the 5'-end to reach 283, 287, 279, 289, 295, 289, and 283 bp, respectively. The vectors for detection of promoter activity were constructed with GUS as a reported gene and used to transform P. quinquefolium callus and tobacco leaves. The results showed that seven PqU6 promoter sequences(PqU6-1P to PqU6-7P) were cloned from the gDNA of P. quinquefolium, with the length ranged from 1 246 bp to 1 308 bp. Sequence comparison results showed that the seven PqU6 promoter sequences and the AtU6-P promoter all had USE and TATA boxes, which are essential elements affecting the transcriptional activity of the U6 promoter. The results of GUS staining and enzyme activity test showed that all the seven PqU6 promoters had transcriptional activity. The PqU6-7P with a length of 1 269 bp had the highest transcriptional activity, 1.31 times that of the positive control P-35S. When the seven PqU6 promoters were truncated from the 5'-end(PqU6-1PA to PqU6-7PA), their transcriptional activities were different in tobacco leaves and P. quinquefolium callus. The transcriptional activity of PqU6-7PA promoter(283 bp) was 1.59 times that of AtU6-P promoter(292 bp) when the recipient material was P. quinquefolium callus. The findings provide more ideal endogenous U6 promoters for CRISPR/Cas9 technology in ginseng and other medicinal plants.

Deep neck space infections: a retrospective cohort study of surgical risk factors / 中华耳鼻咽喉头颈外科杂志

Objective: To explore risk factors affecting treatment for deep neck space infections (DNSIs) so as to provide guidance for appropriate early managements. Methods: A retrospective cohort study was conducted on inpatients with DNSIs admitted to the Department of Otolaryngology, Head and Neck Surgery, Affiliated Hospital of Qingdao University from March 2013 to February 2021. Patients were divided into surgical and non-surgical groups based on whether they had surgery or not. Information collected included demographic data, disease-related signs and symptoms, treatment history, systemic comorbidities, imaging data and laboratory indicators. Hypothesis testing, univariate Logistic regression and multivariate Logistic regression were used for data processing. Resuts A total of 61 patients were included, including 37 males and 24 females, aged 6-96 years. There were 35 cases (57.4%) in the surgical group and 26 cases (42.6%) in the non-surgical group. Multivariate analysis showed that risk factors for surgery as followings: neck dyskinesia (OR=0.03, 95%CI: 0.00-0.24), dysphagia (OR=0.10, 95%CI: 0.02-0.72), serum white blood cell count≥16.74×109/L (OR=1.18, 95%CI: 1.01-1.39) and interspace gas (OR=0.03, 95%CI: 0.00-0.30). Conclusion: Clinicians should be alert to these risk factors for surgery in the course of treatment and timely surgical treatment for patients who meet the conditions.

Ferroptosis in laryngeal squamous cell carcinoma and its regulation by M2 macrophage-derived exosomes / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate ferroptosis in laryngeal squamous cell carcinoma (LSCC) and its regulation by M2 macrophage-derived exosomes. Methods: LSCC and adjacent noncancerous tissue samples were collected from 32 patients treated in the Department of Otorhinolaryngology, Head and Neck Surgery of the Second Affiliated Hospital of Harbin between September 2018 and April 2021, including 26 males and 6 females, aged 43-79 years. The expressions of ferroptosis marker glutathione peroxidase 4(GPX4) in LSCC and adjacent noncancerous tissues were detected by immunohistochemistry and reverse transcriptase-polymerase chain reaction(RT-PCR). The correlations between GPX4 expression and clinicopathological factors in LSCC were analyzed. Biological changes of TU212 cells after treated with ferroptosis-induced agent erastin were detected by transmission electron microscope, cell counting kit-8(CCK-8), clone test, reactive oxygen species(ROS), malondialdehyde(MDA), glutathione(GSH), JC-1, RT-PCR and western blot. Exosomes were isolated from the supernatant of M0/M2 macrophages (M0-exos/M2-exos) and co-incubated with erastin-treated TU212 cells to detect the change of ferroptosis in cells of each group. The data were analyzed by SPSS software of version19.0. Results: GPX4 expression in LSCC tissues was significantly higher than that in adjacent noncancerous tissues (2.04±0.65 vs. 0.99±0.09, F=30.36, P<0.001), and was closely related to T stage and clinical stage (Ⅰ-Ⅱvs.Ⅲ-Ⅳ: 1.75±0.39 vs. 2.18±0.71, F=2.25, P<0.05; T1-2 vs. T3-4: 1.71±0.42 vs. 2.20±0.69, F=2.06, P<0.05). In TU212 cells treated with erastin, mitochondrial crest became smaller, membrane density increased, proliferation rate decreased, intracellular ROS level increased, mitochondrial membrane potential depolarized, GSH content decreased, intracellular MDA level increased and expressions of GPX4 mRNA and protein decreased. Change of M0 into M2 macrophages was induced by IL-4 stimulation. When erastin-treated TU212 cells were incubated with M2-exos, cell proliferation was partially restored and GPX4 expression was enhanced, and also with the recoveries of levels of ROS, MDA and GSH (all P<0.05). Conclusions: Ferroptosis is one of the cell death ways of LSCC. M2-exos may inhibit ferroptosis of LSCC cells.

Nursing Grading Evaluation System for Patients with Stroke at Home: Construction, Validity and Reliability / 中国康复理论与实践

Objective:To construct a nursing grading evaluation indicator system for patients with stroke at home to provide a basis for graded care. Methods:Based on literature analysis, qualitative interviews and panel meeting, an evaluation indicator system was preliminarily drafted. June to September, 2018, the indicator system was consulted to experts three times. It was used to evaluate 210 patients with stroke at home from September to December, 2018 to test its reliability and validity. Results:The effective recovery rates of the three times of consultation were 85.00%, 89.47% and 100%, and the authority coefficient were 0.878, 0.879 and 0.879. The indicator system included four first-level indicators, 30 second-level indicators and 120 items. The scale-level content validity index (CVI) universal agreement was 0.733, average CVI was 0.927; the item-level CVIs were 0.83 to 1.00. For the overall level, the Cronbach's α coefficient was 0.928, and the Guttman Spilt half-factor coefficient was 0.794. For the dimensions, the Cronbach's α coefficient and the Guttman Spilt half-factor coefficients were all above 0.700. The correlated coefficients inter-evaluator of each indicators were 0.492 to 0.963 (P< 0.05). Conclusion:The nursing grading evaluation indicator system for patients with stroke at home is simple and operable, with satisfactory reliability and validity to grade the level of care for patients with stroke at home.

HDL Coupled Multimeric Micelles Co-delivery of siRNA and Doxorubicin for Cell Uptake and Reversal of Multidrug Resistance / 中国药学杂志

OBJECTIVE: To prepare siRNA/HDL modified Dox/micelle multimeric polymer(siRNA/HDL-Dox/micelle) by using HDL as a siRNA carrier and a targeting ligand and to realize the effective co-delivery of siRNA and antitumor drug. METHODS: HDL was incubated with chol-siRNA to prepare siRNA/LDL complex, then coupled with Dox/micelle to form siRNA/HDL modified Dox/micelle multimeric polymer (siRNA/HDL-Dox/micelle). The particle size and stability were investigated in different medium. HepG2/ADM with P-glycoprotein(P-gp) over-expressed were used to study the cell uptake, sub-cellular localization and anti-tumor efficacy in vitro. The ability of siRNA to silence target genes at mRNA and protein level was examined by RT-PCR and Western blot. RESULTS: HDL exhibited an efficient binding ability for siRNA and protected siRNA from RNase. The size and surface morphology of siRNA/HDL-Dox/micelle confirmed by TEM showed that most of the micelles were compact and spherical, exhibited a narrow size distribution and good dispersion. The particle size and Zeta potential increased with increasing incubation time in pH 5.3 PBS. The siRNA was efficiently delivered into the cells by encapsulation into HDL, and the expression of P-gp is effectively down-regulated at the mRNA level and the protein level, thereby increasing the accumulation of intracellular Dox and enhancing the antitumor activity. CONCLUSION: siRNA/HDL-Dox/micelle could effectively deliver siRNA and Dox into tumor cells, thereby exerting gene silencing, reversing tumor drug resistance and enhancing anti-tumor effect.

Effect of Baihe Gujin Tang on LPS-induced Acute Lung Injury in Mice Based on Nrf2/Keap1/p62 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the protective effect and mechanism of Baihe Gujin Tang on lipopolysaccharide induced acute lung injury (LPS-ALI). Method:KM mice were randomly divided into 5 groups:blank control group, model group, dexamethasone 0.002 g·kg-1 group, Baihe Gujin Tang (0.417, 1.25 g·kg-1) group. Except for the blank control group, the other groups were given LPS to induce the mouse ALI model. Except for the blank control group and the model group, the other groups were continuously given intragastric administration for 7 days on the 1st to 7th days before modeling. The lung tissue of the mice was taken 6 h after modeling, and the wet/dry mass ratio (W/D) of the left lung was measured. The serum levels of superoxide dismutase(SOD), malondialdehyde (MDA),reactive oxygen species (ROS)and nitric oxide (NO) were detected in the mice. Thepathological changes of the lung tissues were observed by hematoxylin-eosin(HE) staining. The expression levels of nuclear factor E2 related factor 2(Nrf2), Kelch-likeECH-associated protein 1 (Keap1), p62 and autophagy associated proteinsLC3Ⅱ proteins in the lung tissues were detected by Western blot. Result:Compared with the blank control group, the W/D of the model group was significantly increased (PPPP-1 group was significantly lower (P-1 group and dexamethasone group were able to significantly inhibited MDA levels in serum (PPPPPPConclusion:Baihe Gujin Tang has obvious protective effect on LPS-ALI mice, and its mechanism may be related to the regulation of Nrf2/Keap1/autophagy feedback loop.

Renoprotection Provided by Dipeptidyl Peptidase-4 Inhibitors in Combination with Angiotensin Receptor Blockers in Patients with Type 2 Diabetic Nephropathy / 中华医学杂志(英文版)

Background@#Treatment with the dipeptidyl peptidase-4 inhibitors (DPP4i) and angiotensin receptor blockers (ARBs) in patients with type 2 diabetic nephropathy (DN) has not been well characterized. This study aimed to assess the renoprotection of this combined treatment in DN patients.@*Methods@#A total of 159 type 2 DN patients from 2013 to 2015 were enrolled retrospectively from a prospective DN cohort at the National Clinical Research Center of Kidney Diseases, Jinling Hospital (China). Fifty-seven patients received DPP4i and ARB treatment, and 102 patients were treated with ARBs alone. All patients were followed up for at least 12 months. Statistical analyses were performed using Stata version 12.0.@*Results@#There were no significant differences at baseline for age, sex, body mass index, duration of diabetes, fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and estimated glomerular filtration rate (eGFR) between the two groups. Antihypertensive and antidiabetic medication use was similar in each group except calcium channel antagonists (P = 0.032). No significant changes in FBG and HbA1c were observed in the two groups after treatment. The eGFR decreased slower in the DPP4i + ARB group than in the ARB group at 12 months (Δ12 months: -2.48 ± 13.86 vs. -6.81 ± 12.52 ml·min·1.73m, P = 0.044). In addition, proteinuria was decreased further in the DPP4i + ARB group than in the ARB group after 24 months of treatment (Δ24 months: -0.18 [-1.00, 0.17] vs. 0.32 [-0.35, 0.88], P = 0.031). There were 36 patients with an eGFR decrease of more than 30% over 24 months. After adjusting for FBG, HbA1c, and other risk factors, DPP4i + ARB treatment was still associated with a reduced incidence of an eGFR decrease of 20% or 30%.@*Conclusions@#The combined treatment of DPP4i and ARBs is superior to ARBs alone, as evidenced by the greater proteinuria reduction and lower eGFR decline. In addition, the renoprotection of DPP4i combined with ARBs was independent of glycemic control.

Metabolomic approach to evaluating the effect of the mixed decoction of kelp and licorice on system metabolism of SD rats / 药学学报

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.

Metabolomic approach to evaluating the effect of the mixed decoction of kelp and licorice on system metabolism of SD rats / 药学学报

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.

Synthesis and anti-tumor activity of baicalin-metal complex / 药学学报

Chelating ligand method has been used to synthesize baicalin-metal (Ni2+, Co2+, Cu2+) complexes (BMC). The composition and structure of BMC were characterized by the element analysis, ultraviolet spectrum (UV), infrared spectrum (IR), mass (MS) and thermal gravitational analysis (TGA). MTT was used to analyze the effects of BMC on SMMC-7721 cell proliferation. PI staining method and Annexin-V/FITC double staining method were used to analyze the effects of BMC on the cell cycle and apoptosis of SMMC-7721 cell. Fluorescence quantitative RT-PCR was used to analyze the expression of BMC on Bcl-2 gene and Bax mRNA, flow cytometry was used to analyze BMC on the expression of Bcl-2 protein and Bax protein. The antineoplastic activity and mechanism of action of BMC was explored comprehensively. The results showed that three new kinds of BMC (molar ratio of 2 : 1) were successfully prepared, the complexes molecular formula are: Na2Ni(C21H16O11)2 x 10H2O, Na2Co(C21H16O11)2 x 8H2O and Na2Cu(C21H16O11)2 x 8H2O. According to the results of cell cycle and apoptosis detection, BMC stopped cells at G0/G1 phase to S phase and G2/M phase. Gene and protein detection showed that under the given concentration and time, BMC can downregulate the expression of Bcl-2 gene in SMMC-7721 cells, and significantly decrease the expression of Bcl-2 protein, at the same time, with the increase of expression of Bax gene, the Bax protein's expression increased significantly. Which indicates that BMC restrain cell proliferation and cell apoptosis by stopping cell cycle, reducing the expression of Bcl-2 and increasing that of Bax; The anti-tumor activities of three kinds of complexes were: baicalin-copper (BC-Cu) > baicalin-cobalt (BC-Co) > baicalin-nickel (BC-Ni) > baicalin (BC), showing the dose-response relationship.

Study on preparation of deoxyrhaponti-beta-cyclodextrin super-molecular inclusion complex and performance of inclusion complex / 中国中药杂志

To prepare beta-cyclodextrin (beta-CD)-deoxyrhaponti inclusion complex by the homogeneous method, and characterize the inclusion complex with ultraviolet-visible spectrum and fluorescence spectroscopy, in order to determine the inclusion rate, the ratio of subject-object, the binding constant of supra-molecular system and the thermodynamic function. The results showed that the designed method was so rational that the inclusion complex was successfully prepared. The ultraviolet-visible spectrophotometry method was adoptedto determine the inclusion rate of 38%, and the ratio of subject-object of 2: 1. The thermodynamic parameters of this inclusion complex: deltaH(0), deltaS(0) was all smaller than zero, which indicated that the main acting forces generated by the inclusion complex were hydrogen bonding and Vander Waals' force. deltaG(0) < 0 and deltaG(0) were directly proportional to the reaction temperature, which suggested that the reaction would spontaneously increase with the temperature rise. The polarization fluorescence method was used to quantitatively prove the non-covalent inclusion complex generated during the interaction between beta-CD interacting with DES. The results could also provide reference for studies on cyclodextrin supra-molecular system.

Change in ambulatory arterial stiffness index with advancing age and the correlation with 24 hours pulse pressure in healthy individuals / 中华心血管病杂志

<p><b>OBJECTIVE</b>Ambulatory arterial stiffness index (AASI) has been recently proposed to reflect the dynamic relation between diastolic and systolic blood pressure throughout the whole day. The aim of our study was to investigate the change in AASI with advancing age and the correlation with 24 hours pulse pressure (24 h PP) in healthy individuals.</p><p><b>METHODS</b>246 healthy subjects [mean age (59.7 +/- 14.6) years, women 38.6%] underwent 24 hours ambulatory blood pressure monitoring (ABPM) in normal life style. The blood pressure recordings, heart rate (HR), mean arterial pressure (MAP) and pulse pressure (PP) were analyzed simultaneously by computer for every 30 minutes during 6:00 am-22:00 pm and every 60 minutes during 22:00 pm-6:00 am. Using all the blood pressure recordings, we plotted diastolic against systolic blood pressure from each individuals and calculated the regression slope. AASI was derived from 1 minus this regression slope.</p><p><b>RESULTS</b>In 246 healthy individuals, AASI increased with age. Among the healthy individuals, the 95th percentile of AASI was 0.56, the upper boundary of the 95% prediction interval of AASI in relation to age were 0.49 at 20 - 39 years, 0.59 at 40 - 59 years, 0.69 at 60 - 79 years, 0.79 at > or = 80 years. The correlation coefficient between AASI and 24 h PP was 0.497 (P < 0.01). AASI linearly increased with age in healthy individuals, whereas the relation between pulse pressure and age was curvilinear.</p><p><b>CONCLUSIONS</b>AASI as a index reflecting blood pressure relationship, manifested the corresponding change with advancing age. The correlation between AASI and traditional index 24 h PP indicated AASI as a new measure of arterial stiffness.</p>

Effects of the extracts from decoction for resuscitation and its component herbs on PGI2, TXA2 and NO release from rat vascular endothelial cells under hypoxia in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the effect of the extracts from Decoction for resuscitation (DRE) and its component herbs on prostacyclin (PGI2), thromboxane A2 (TXA2) and nitric oxide (NO) release from rat vascular endothelial cells under hypoxia.</p><p><b>METHOD</b>After treatment with the extracts from DRE and its component herbs, the contents of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha), thromboxane B2 (TXB2) as well as nitrite (NO), which were degradation products of PGI2, TXA2 and NO respectively, in culture medium of rat vascular endothelial cells under hypoxia were measured with radioimmunoassay and Griess Reaction.</p><p><b>RESULT</b>Compared with the control group, the results indicated that DRE, prepared licorice root extract (LE), dried ginger extract (GE), aconite root extract (AE), extracts of aconite root and prepared licorice root (ALE), extracts of aconite root and dried ginger (AGE) increased significantly the content of 6-keto-PGF1alpha and the ratio of 6-keto-PGF1alpha/TXB2, but had no effect on the content of TXB2 in culture medium of rat vascular endothelial cells under hypoxia. The content of 6-keto-PGF1alpha in the DRE group was higher than that in the groups of LE, GE, AE, ALE, AGE. The ratio of 6-keto-PGF1alpha/TXB2 in the DRE group was higher than that of the groups of GE, AE, ALE. Compared with the control group, DRE, LE, GE, AE, ALE, AGE increased significantly the content of NO2- in culture medium of rat vascular endothelial cells under hypoxia. Moreover, the content of NO2- in the DRE group was higher than that of the groups of GE, AE, ALE.</p><p><b>CONCLUSION</b>The results suggested that DRE increased significantly the content of PGI2 and the ratio of PGI2/TXA2 as well as the content of NO. The effect of DRE on the parameters in culture medium of rat vascular endothelial cells under hypoxia was better than that of the extracts from its component herbs.</p>

Construction of the mutants of rice nonspecific lipid transfer protein and expression comparison in two kinds of thioredoxin fusion expression vectors / 生物工程学报

Five structural important residues of rice nonspecific lipid transfer protein LTP110 were mutated by site-directed mutagenesis. Sequence results showed that they were all mutated successfully. After trying various E. coli expression systems, thioredoxin fusion expression system was found to be a proper system to express wild type and mutant LTP110. cDNA sequences encoding wild type LTP110 and the mutants Y17A, P72L, R46A, D43A, C50A were cloned into two kinds of thioredoxin fusion expression vectors. The expression results were compared. In pTrxFus/GI724 expression system, wild type LTP110 and the mutants Y17A, P72L, R46A could be expressed at low level while D43A and C50A could not be expressed normally; in pET32a(+)/BL21 (DE3) trxB- expression system, wild type LTP110 and all mutant proteins could be expressed very well and the levels were higher than that in pTrxFus/GI724 system. LTP110 fusion protein expressed in pET32a(+) vector was purified and its activity was checked by fluorescence labeled fatty acid. Results indicated that the recombinant LTP110 fusion protein has lipid binding activity. This work provides good basis for the further study.