Phosphoethanolamine N-methyltransferase (PEAMT) is a key enzyme that catalyzes the synthesis of phosphocholine, which is an important precursor of phosphatidylcholine and glycine betaine. A 1249bp 5'-flanking region of phosphoethanolamine N-methyltransferase gene was isolated by anchored PCR, based on the cDNA sequence of PEAMT from halophyte Salicornia europaea. The transcription start site was identified as A and localized at 301bp upstream of the ATG according to the results of RLM-RACE. In SePEAMT promoter region, many potential cis-acting elements were predicted by PlantCARE and PLACE programs. Aside from the basal transcriptional elements TATA-box and CAAT-box, some stress-responsive motifs such as ABRE, HSE and LTR were found. In addition, some pollen-specific activation-related elements were also present in this region. Binary expression vector was constructed by fusing SePEAMT promoter with GUS gene and designated as pPro. The pPro was transferred into tobacco by Agrobacterium-medicated transformation and transient GUS expression analysis indicated that SePEAMT promoter could drive strong GUS expression.

Temporal and spatial expressions of ?-catenin investigated in the hair follicle and epidermis of the hoof periphery in bovine embryonic development. IHC (immunohistochemical method) was applied to qualitatively detect the temporal and spatial expressions of ?-catenin. ?-catenin was detected in suprabasal, epidermal basal layer, placode, hair bud in early phase(E68～93),and expressed strongly in epidermal basal layer, placode, and hair bud, in suprabasal expressed less strongly; in metaphase(E94～184), ?-catenin was detected in epidermis, hair peg, and in suprabasal, epidermal basal layer, hair follicle bulge, inner root sheath, outer root sheath, follicular infundibulum expressed less strongly; in late phase(E184～225), ?-catenin expressed weakly in epidermal basal layers, while expressed strongly in epidermal keratinocytes. The result suggested that ?-catenin plays an important role in hair follicle morphogenesis in the periphery of bovine hoof in bovine embryos.

This study is to establish an UPLC fingerprint of Resina Draconis from different manufacturers, which can provide a comprehensive evaluation for its quality control. The analysis was performed on a Phenomenex Kinetex 2.6 μ C18 100A column by agradientelution program with acetonitrile-water as mobile phase at a flow rate of 1.7 mL x min(-1). The column temperature was 40 degrees C and the detection wavelengthwas 280 nm. The fingerprints of 18 batches of Draconis Resina were further evaluated by chemometrics methods including similarity analysis (SA), hierarchical clustering analysis (HCA) and principal component analysis (PCA). As a result, there were 15 common peaks, 13 of which had been identified by LC-Q-TOF MS, and the similarity degrees of 15 batches of the samples was more than 0.9, and the samples were divided into 4 clusters by their quality difference. The method is reproducible, simple and reliablethat it can be used for quality control and evaluation of Resina Draconis from different manufacturers.
Chromatography, High Pressure Liquid ; methods ; Dracaena ; chemistry ; Drugs, Chinese Herbal ; analysis ; Principal Component Analysis ; Quality Control

Objective To study the feasibility of a novel probe 99Tcm-HYNIC-2(poly-(ethylene glycol),PEG) 4-Dimer (Dimer:E-[c (RGDfK) 2]) as a potential imaging agent for integrin αv β3 positive tumors,and also to observe the influence of an angiogenesis inhibitor,endostar,on the biodistribution and tumor uptake of the tracer in tumor bearing nude mice.Methods The expression of integrin αv β3 in human glioma cells U87MG was determined with immunofluorescence staining before and after treatment with endostar.99Tcm-HYNIC-2PEG4-Dimer was prepared and administered in U87MG tumor bearing mice in 6 h after either administration of endostar (200 μl) or saline (control group) and then biodistribution study was performed.Other 16 mice were divided into endostar treated group (20 mg/kg) and control group (saline) and then gamma imaging was performed in the two groups.Statistical significance of differences between the two groups was assessed using two-sample t test.Results Radiochemical purity of 99Tcm-HYNIC-2PEG4-Dimer was exceeded 95％.The expression of integrin αvβ3 in U87MG cell was high and gradually decreased after treatment with endostar.There was a negative dose-effect relationship between the dose of endostar and the expression of integrin αvβ3 with the peak effect at the dose of 400 μg/ml.The distribution study in vivo showed that the tracer uptake of U87MG tumors was high,but it decreased after injection of endostar.At 90 min,the ％ID/g of endostar and control groups were 1.50±0.08 and 6.27±0.33,respectively (t =40.23,P＜0.05).The average T/NT ratios of 99Tcm-HYNIC-2PEG4-Dimer uptake in the endostar and control groups were 1.02±0.11 and 2.58±0.36,respectively (t =10.25,P＜0.05).The integrin αv β3 positive expression ratios of tumor in endostar and control groups were (33.1 ±2.7) ％ and (81.5±3.2) ％,respectively (t =32.60,P＜0.05).Conclusions The novel probe 99Tcm-HYNIC-2PEG4-Dimer may be a promising radiotracer for integrin αvβ3-positive tumor imaging.It may be used for monitoring the therapeutic effect of endostar and may be potentially used for screening the candidates of anti-angiogenesis therapy.

Objective Metabonomics method based gas chromatography-mass spectrometry (GC/MS)were used to analyze the urine samples of patients with hyperlipidemic pancreatitis (HLP) to describe the characteristics of metabolism changes of HLP,identify potential biomarkers,and investigate the role of metabonomics study in the management of AP.Methods 24 patients of HLP and 40 age,sex matched volunteers were enrolled and their urine samples were collected.The urine samples underwent preparation,derivation and GC/MS analysis,Orthogonal-Projection to Latent Structures-Discriminant Analysis (OPLS-DA)were performed to detect the metabolic profile difference between the HLP and control group.Results HLP patients can be precisely distinguished from healthy controls.21 metabolites (credibility ＞ 700 ) were identified using the reference compounds available in the libraries of NIST and Wiley.It was identified that levels of nicotinic acid,aconitie acid,citric acid,hippurie acid,hydroxyphenylacetic acid,hydroxyphenylpropionicacid were decreased,while the levels of tryptophan,tyrosine,tyramine,16-hexadecanoic acid,18octadecanoie acid were increased.It was also suggested that there was change in tricarboxylic acid cycle and gut bacterial flora,as well as fat metabolism and metabolism of amino acid.Conclusions There are differences between healthy controls and HLP patients in the term of GC/MS metabolic profiling,and the biomarkers in the metabolites could be found through metabonomics analysis,and the mechanisms of the metabolic changes could be explored.It was noted that the research of metabolites in the urine samples may be a useful tool to help diagnose and understand the pathogenesis of HLP.Metabonomics analysis is a promising research method.

Objective To study to take the oxidized Mannan?modified 786?0 in renal clear cell carcinom as tumor cells antigen to sensitize Dendrit?ic cells(DC)and to observe the its killing effect on renal clear cell carcinoma of CTLs induced. Methods Getting the peripheral blood mononucle?ar cells from the volunteers,and then to be stimulated to turn to be maturation by GM?CSF and IL?4 in vitro. Taking the clear renal carcinoma cell as the tumor antigen,and then making it to be modified by oxidized Mannan to acquire the tumor cell vaccine.Experimental groups include:blank group:DC?PBS group,control group:control?DC?786?0,experimental group:DC?Ox?Mannose?786?0 group. Taking the flow cytometry to detect the changes of DC phenotype,then taking the ELISA to detect the sencretion levels of supernatant of IL?12 of DC,then taking the CCK to detecte the cytotoxicity of lymphocytes(CTL)induced by DC of these experiment groups. Results Results by flow cytometry:the mature phenotype of DCs sensitized by Ox?Mannose?786?0 group included CD80,CD83,CD86 and HLA?DR expressed significantly higher than the other groups. As well as the secretion levels of IL?12. Meanwhile the cytotoxicity activity of lymphocytes(CTLs)induced by DCs which are sensitized by Ox?Mannose?786?0 increased more significantly than the other groups. Conclusion Glycosylated Antigens can be more effective in sensitizing antigen?presenting cells DC,and stimulating them to be maturation,while the killing effect to tumor cells also have noticeably improved.

Objective To investigate the correlation of T cell subgroups and natural killer (NK) cell's activity level of peripheral blood of the patients with refractory lymphoma. Methods Flow cytometry was applied to detect T cell subgroups' level and NK cell's activity of peripheral blood in 60 early cure lymphoma patients with chemotherapy before and 20 normal controls , after chemotherapy follow-up they were divided into 30 cases of difficult cure group and 30 cases of effective group. Results Compared with the normal controls, CD+4, CD+4/CD+8 and NK cell in lymphoma patients with chemotherapy before decreased (30.17±8.63 vs 46.52±1.39, t =12.218, P ＜0.05; 0.86±0.45 vs 1.64±0.05, t =11.225, P ＜0.05; 12.39±7.08 vs 19.29±0.84,t =6.365, P＜0.05), while CD+3 and CD+8 cell increased (76.14±10.71 vs 70.48±1.44, t =-3.439, P＜0.05;40.28±14.03 vs 28.35±0.73, t =-5.625, P ＜0.05). Compared with effective group, CD+4 CD+4/CD+8 and NK cell in difficult response group with chemotherapy before decreased (27.70±7.81 vs 33.13±8.82, t =2.163, P =0.036;0.67±0.27 vs 1.10±0.52, t =3.272, P =0.003; 9.87±6.60 vs 15.40±6.58, t =2.771, P =0.008), while CD+3 and CD+8 cell increased (79.67±8.18 vs 71.91±12.00, t =-2.540, P =0.015; 44.70±13.99 vs 34.98±12.41, t =-2.416,P =0.020). Conclusion The detection of T cell subgroups' level and NK cell' s activity in early lymphoma patients before chemotherapy may play a role to diagnose and predict the outcome of refractory lymphoma patients.

Objective To investigate the common chromosome abnormalities in the patients with multiple myeloma and the relationships of cytogenetic abnormalities and clinical features. Methods The interphase fluorescence in situ hybridization (I-FISH) analysis method was designed to detect RB1-/13q14-and 14q32 rearrangements in 49 MM patients. The statistic value of its effect on clinical features were determined. Results FISH disclosed 14q32 translocations in 26 of the 40 (53.1%) patients. 25 out of the 49 (51.02 %) cases were found with deletion of chromosome 13q14 included del(RB1) in 9 (18.4 %) and del(13q14.3) in 18 (36.7 %). 13q14 deletion and 14q32 translocation were simultaneously observed in 18 (36.7 %) cases. Spearman correlation analysis were found associated of 14q32 rearrangement with the percentage of plasma cells in bone marrow (r=0.316, P=0.27). Conclusion The frequency of 13q14 deletion and 14q32 gene translocation in multiple myeloma are high. There is a significant correlation between the presence of 14q32 translocations and chromosome 13 abnormalities in MM patients. The percentage of 14q32 translocation in plasma cells was increased significantly. The 14q32 translocation is an independent prognostic factor.

Protein phosphorylation is one of the most common post-translational modifications (PTMs)in various organisms,which plays critical roles in the regulation of intracellular biological processes,such as cell prolifera-tion,signal transduction,metabolismis and tumorigenesis.However,the low abundance of phosphoprotein in the biological systems poses significant challenges of current analytical techniques.In order to further understand the phosphoproteomics,the roles of phosphorylated proteins in life process,discovery of biomarkers,diagnosis and treatment of disease,enrichment strategies of high efficiency have been developed,including the design of new nanomaterials and combination of a variety of analytical methods,et al.In this paper,we reviewed the develop-ment of enrichment strategies for phosphoproteomics and application of phosphoproteomics in disease.

Abstract: Objective　This article summarizes the clinical characteristics and diagnosis and treatment experience of an elderly patient infected with Omicron variant BA.5.1.3 of COVID-19 in Hainan Province. Methods　The clinical data and treatment of an elderly patient infected with Omicron variant BA.5.1.3 of COVID-19 admitted to Haikou designated hospital on August 15, 2022 were retrospectively analyzed. Results　A 107-year-old female patient was admitted to the hospital with "fever and cough for 1 day". Two of her family members have infected with COVID-19. The patient initially developed fever, accompanied by cough, expectoration, a little white sticky sputum, accompanied by sore throat, muscle pain, fatigue. Nucleic acid test was positive in throat swab, indicating Omicron variant BA.5.1.3 infection. The patient was diagnosed as mild COVID-19 and treated with antiviral therapy, Chinese medicine conditioning, anticoagulation, electrolyte disorder regulation and symptomatic treatment for 9 days. The patient's clinical symptoms were relieved, and she was cured and discharged after two negative nucleic acid tests. One week later, the patient recovered well. Conclusions　Omicron variant BA.5.1.3 is highly infectious, and comprehensive treatment such as antiviral treatment and traditional Chinese medicine treatment has achieved good efficacy. For elderly patients, attention should be paid to maintaining the stability of organ function and internal environment, which is helpful to improve the prognosis of patients.