Objective To construct pET21a-sBAFF by cloning the extracellular regions of 134-285 amino acids of BAFF, a member of human TNF family, and then express the gene in prokaryotic cells and purify the expressed product.Methods cDNA of K562 cell line was used as the template to amplify sBAFF gene to construct pET21a-sBAFF.Expression of sBAFF in E.coli BL21 was induced by IPTG, and the expressed proteins were assayed by SDS-PAGE.The bacteria were analyzed by sonication, and the target proteins mainly existed as inclusion bodies.Then sBAFF was purified by Ni2 +-IDA affinity chromatography.SDS-PAGE electrophoreses displayed that the expressed sBAFF migrated with a relative molecular weight of 18000.ResuIts The induction parameters such as temperature and inducing time were optimized.The target protein accounted for 38.59%of the total bacterial proteins.After refolding, 38.14% of sBAFF proteins were polymerized as an active trimer.The dimer form of sBAFF, which is representative of wrongly refolded product, accounted for very few.ConcIusion The expression and purification of BAFF which formed active trimer after refolding pave the way for its further function study and provide a novel approach for the development of BAFF-targeted therapeutics for autoimmune diseases.

Objective:To study the biological characteristics of condylar chondrocytes isolated from mechanical stress stimulated TMJ condylar cartilage of rats.Methods:The rat models of mechanical stress stimulated condylar cartilage were made by class III orthope-dic force for 14 days.The cartilage from control and experimental rats were observed in gross view.The cell harvest rate and viability were examined,proteomic analysis was performed.Results:After stress stimulation the thickness,the elasticity of condylar cartilage and number of chondrocytes were significantly reduced.The weight of mandibular cartilage was decreased(P<0.01).The harvest rate and the viability rate of the cells were decreased(P<0.01).The chondrocytes in the experimental group appeared to be elongated. Proteomic analysis showed that stress-related proteins,signal transduction proteins were up-regulated;cytoskeleton proteins,cell prolif-erative proteins were down-regulated.Conclusion:Mechanical stress stimulation of condylar cartilage may result in biological activity and protein changes of the condylar cartilage chondrocytes.

This study is to investigate the apoptotic induction effect of the combination of diosgenin and TNF-related apoptosis-inducing ligand (TRAIL) on non-small-cell lung cancer cell line A549 by using the Chou-Talalay method, and observe the mechanism of the combination. The apoptotic effect of diosgenin or TRAIL alone and their combination on A549 and normal cell line 293T proliferation was measured by MTT assay. Chou-Talalay method was used to evaluate the combination effect. Apoptosis was examined by Hoechst 33342 staining and flow cytometry assay. Western blotting detects the expression of apoptosis-associated proteins. Diosgenin or TRAIL alone can inhibit proliferation ofA549 in a concentration-dependent manner. According to the Chou-Talalay method, when f(a) = 0.1, CI > 1, when f(a) > 0.1, CI < 1. Combined with TRAIL, the IC50 of diosgenin decreases from 21.864 to 14.810 micromol x L(-1) (P < 0.05) on A549 cells. But for 293T cells, IC50 of diosgenin does not change significantly. As with Hoechst 33342 staining and flow cytometry assay, the apoptosis ratios also increased in the combination group. At protein expression level, combination-treated group displays increased Caspase-8, Caspase-9, Bid, Caspase-3 activation and PARP cleavage, significantly decreased Bcl-2 and increased Bax expression, and MAPK pathways were activated. The combination of diosgenin and TRAIL has synergistic effect on A549 cells.

Objective To synthesize 99Tcm-TP5-3 and evaluate its biodistribution and kinetics as a molecular probe for the detection of apoptosis,and evaluate tumor apoptosis after a single dose of paclitaxel chemotherapy in MDA-MB-231 breast tumor model.Methods TP5-3 was labeled with 99Tcm directly,and analyzed with HPLC.The radioactivity in tissues was measured and expressed as ％ID/g and T/NT (tumor/muscle).The mice bearing MDA-MB-231 breast tumor were divided into two groups:the treatment group which was given a single dose of paclitaxel (40 mg· kg-1,via tail vein),and the control group which was injected with the same volume of normal saline.After therapy,99Tcm-TP5-3 was injected via tail vein in both groups (100 μ1 for each mouse).MicroSPECT/CT was performed at 3 h postinjection.Radioactivity in different tissues was determined after imaging.Apoptotic cells were measured with flow cytometry.The morphological changes of the apoptotic cells were observed by light microscopies.One-way analysis of variance,two-sample t test and linear correlation analysis were used to analyze the data.Results The radiolabeling efficiency was ＞ 95％ and the radiochemical purity of 99Tcm-TP5-3 was (96.0± 1.5)％ at room temperature for 4 h.The predominant uptake was found in the kidneys at 30 min postinjection ((8.48± 1.07) ％ID/g),with rapid tracer clearance from the circulation.By comparison with activity at 5 min postinjection ((13.74± 4.21) ％ID/g),85％ of the initial activity reduced in blood at 4 h ((2.07±0.35) ％ID/g; F=11.310,P＜ 0.05).99Tcm-TP5-3 was mainly accumulated in the kidneys,liver and stomach,and excreted via the kidneys.T/NT in the treated group was 4.21±0.06,which was significantly higher than that of the control group (1.57±0.67; t =12.820,P＜0.05).The radioactivity of tumor tissue in the treatment group was much higher than that in the control group (4.82±0.54) ％ID/g vs (1.44±0.38) ％ID/g,t=0.679,P＜0.05).The tumor uptake of 99Tcm-TP5-3 in the treatment group positively correlated well with the apoptotic cells (r =0.985,P＜0.05).Histopathology further confirmed that a large number of apoptosis had occurred in the tumor after paclitaxel treatment.Conclusion 99Tcm-TP5-3 appears to have potential to be a useful molecular probe for imaging tumor cell apoptosis.

Hirudin is one of the most potent anti-coagulant protein ever found, and its C-terminus is a key domain for inhibiting thrombin.In order to enhance its specificity,a novel anti-coagulant protein was constructed via fusing the C-terminus of hirudin to Annexin V, which was expected to sustain both anti coagulant activity and phorspholipid affinity. The structure of the designed protein was predicted with both molecular mechanics and dynamics. Molecular dynamics was adopted to simulate the docking interaction between the fusion protein and thrombin. The results showed the inhibitory activity of the fusion protein to thrombin.

Objective To study the prevalence of hypertension in She Chinese population of Fujian province and its epidemiological characteristics. Methods Using random sampling method to take advantage of number table, we select a sample of 5350 people who were conducted a questionnaire survey and measured weight, height, blood pressure and other indicators. Results The prevalence of hypertension in She Chinese population of Fujian province was 36. 09%, including undiagnosed number of 1374 cases. The main risk factors of hypertension were age,the level of education, BMI,saltintake. Smoking was not significant with hypertension. The prevalence rate of hypertension among people over 60 years was 63.10%, people comsumed above 8 grams of salt per day had higher pervalence than that in the goup which comsumed below 6 grams or between 6 grams to 8 grams of salt per day. Conclusions The prevalence of hypertension in She had grown rapidly. The She Chinese population should change their lifestyle and hypertension education should perform in this population.

Object:To explore the clinical effect of periacetabular osteotomy (PAO) in the treatment of developmental dysplasia of hip (DDH).Methods:The clinical data of 305 patients with DDH admitted to Tangshan Second Hospital from January 2012 to June 2018 were analyzed retrospectively. All patients were treated with PAO alone. There were 123 cases of left hip dysplasia, 131 cases of right hip dysplasia and 51 cases of double hip dysplasia (356 hips in total). The patients underwent X-ray examination before and on the second day after operation to compare the acetabular index and the lateral center edge angle (LCEA) and the anterior center edge angle (ACEA) of the acetabulum before and after operation. The ability of daily living and hip joint function were evaluated before and 6 months after operation, and Barthel index and Harris score of hip joint were compared before and after treatment. The measurement data with normal distribution are expressed in xˉ± s, and the paired t test was used for comparison before and after operation. The measurement data of non-normal distribution is expressed by M( Q1, Q3), and the comparison before and after surgery is performed by the rank sum test. Results:On the second day after operation, the LCEA and ACEA of 356 hip joints in 305 patients were greater than those before operation (32.5(20.0,47.5)° vs 8.5(-18.5 23.0)°, 29.0(18.5,52.3)° vs 2.5(-20.8, 24.5)°), while the acetabulum index was lower than that before operation (6.7(-8.4,12.5)° vs 26.8(10.0, 62.3)°), and the differences were statistically significant ( Z values were 51.50, 45.37, 32.22, all P<0.001). After 6 months of follow-up, the Barthel score and Harris score of the hip joint were higher than those before the operation (92.5±1.3) scores vs (65.6±1.5) scores, (96.4±2.5) scores vs (85.1±1.3) scores, and the difference was statistically significant ( t values were 335.56 and 89.70, both P<0.001). Conclusions:PAO can make the acetabulum cover the femoral head well through acetabular transposition, improve the ability of daily living and hip joint function of DDH patients, reduce pain, increase joint range of motion, and correct limb deformities. It is an effective means to treat DDH.

CircRNA is a single-stranded, covalently closed non-coding RNA, whose mechanism mainly involves sponge adsorption of micro RNA (miRNA), regulation of protein transcription and post-transcriptional levels, interaction with RNA-binding proteins and few coding proteins of circRNA. Meanwhile, because of its abundant expression, highly conserved and dynamic changes, circRNAs have promising applications such as becoming biomarkers of diseases, developing circRNA vaccines and gene editing therapies for circRNAs. Cardiovascular diseases are the leading cause of death from diseases worldwide, and existing therapeutic approaches can delay the development of heart diseases, but are still limited by the unknown pathogenesis and therapeutic targets. Many studies have reported the mechanistic link between circRNAs and cardiac diseases, therefore, this review is to explain the progress of circRNA research in cardiovascular diseases and to illustrate the three clinical applications in which circRNAs are currently involved.

Objective To analyze medication rules of traditional Chinese medicine in the treat-ment of thyroid nodules based on real-world data mining technology.Methods Chinese medicine prescriptions for patients with thyroid nodules as clinical first diagnosis in the Department of Thyroid and Breast Surgery in Qingdao Hospital of Traditional Chinese Medicine from August 2018 to August 2023 were collected,and the Traditional Chinese Medicine Inheritance Assistant Platform(V3.0)was used to analyze the medication rules.Results A total of 1 206 traditional Chinese medicine pre-scriptions for the treatment of thyroid nodules were screened,involving 291 traditional Chinese medi-cines;the top five most frequently used drugs were Fritillaria thunbergii,Prunella vulgaris,Bupleu-rum chinense,Poria cocos,and Pinellia ternata;the main function of drugs were clearing heat,toni-fying deficiency,resolving phlegm,activating blood circulation,and regulating qi;the main property of the drugs was cold,the main tastes of drugs were bitter,sweet and spicy,and the meridians in-volved were mainly the lung,liver and spleen meridians;association rule analysis identified 17 high-frequency drug combinations,with Fritillaria thunbergii-Prunella vulgaris as the most frequently oc-curring drug pair;among the 16 association rules,Radix Scrophulariae-Fritillaria thunbergii and Rhi-zoma Cyperi-Bupleurum chinense had the highest confidence level;clustering analysis found 6 core drug combinations.Conclusion The main idea of clinical treatment of thyroid nodules in traditional Chinese medicine is to resolve phlegm and reduce nodules,promote qi circulation and blood circulation,and nourish qi and blood;the common herbal combination of Fritillaria thunbergii and Prunella vul-garis is used to clear heat,resolve phlegm,and reduce nodules,and the basic formula for resolving scrofula is based on Xiaoluo Pills,which resolves phlegm,softens hardened nodules,and reduces nodules.

Objective To analyze medication rules of traditional Chinese medicine in the treat-ment of thyroid nodules based on real-world data mining technology.Methods Chinese medicine prescriptions for patients with thyroid nodules as clinical first diagnosis in the Department of Thyroid and Breast Surgery in Qingdao Hospital of Traditional Chinese Medicine from August 2018 to August 2023 were collected,and the Traditional Chinese Medicine Inheritance Assistant Platform(V3.0)was used to analyze the medication rules.Results A total of 1 206 traditional Chinese medicine pre-scriptions for the treatment of thyroid nodules were screened,involving 291 traditional Chinese medi-cines;the top five most frequently used drugs were Fritillaria thunbergii,Prunella vulgaris,Bupleu-rum chinense,Poria cocos,and Pinellia ternata;the main function of drugs were clearing heat,toni-fying deficiency,resolving phlegm,activating blood circulation,and regulating qi;the main property of the drugs was cold,the main tastes of drugs were bitter,sweet and spicy,and the meridians in-volved were mainly the lung,liver and spleen meridians;association rule analysis identified 17 high-frequency drug combinations,with Fritillaria thunbergii-Prunella vulgaris as the most frequently oc-curring drug pair;among the 16 association rules,Radix Scrophulariae-Fritillaria thunbergii and Rhi-zoma Cyperi-Bupleurum chinense had the highest confidence level;clustering analysis found 6 core drug combinations.Conclusion The main idea of clinical treatment of thyroid nodules in traditional Chinese medicine is to resolve phlegm and reduce nodules,promote qi circulation and blood circulation,and nourish qi and blood;the common herbal combination of Fritillaria thunbergii and Prunella vul-garis is used to clear heat,resolve phlegm,and reduce nodules,and the basic formula for resolving scrofula is based on Xiaoluo Pills,which resolves phlegm,softens hardened nodules,and reduces nodules.