Objective To clone the full length staphylococcal protein A(SPA) gene from Staphylococcus aureus (ATCC6538), and subsequently study the gene structure and antibody binding ability.Methods The full length and the functional region of the SPA gene were cloned into pHisSUMO vector respectively, and expressed in E. coli. The full length and the functional fragment of the SPA protein were detected for antibody binding ability and stability. The functional fragment of the SPA protein fused with SUMO was coupled to the CNBr-activated agarose for antibody purification from rabbit serum. Results A variant of the full length SPA gene was cloned, which has been submitted to GenBank (the accession number is EU695225). Two fusion proteins had the same antibody binding ability as the untagged SPA protein. However, the formers was more stable than the latter at the tested conditions. SUMO-SPA conjugated-agarose kept high efficiency for antibody binding. Conclusion To our knowledge, the full length SPA gene of S.aureus(ATCC6538) is a novel variant. The SUMO tag can improve the stability of the functional region of the SPA protein without damaging the antibody binding ability. This fusion protein has been used for antibody purification successfully.

Objective To investigate the role of neuropeptide trefoil factor 3 (TFF3) in cocaine reward responses and the underlying mechanism.Methods Fifty SD rats were divided into 6 groups including Control group,Cocaine group,Cocaine + TFF3 (0.01 mg/kg) group,cocaine + TFF3 (0.1 mg/kg) group,cocaine + TFF3 (0.5 mg/kg) group and TFF3 (0.1 mg/kg) group,ip,n=7 ～ 9),and were treated with TFF3 / saline (ip),30 min later,rats were injected with cocaine (10 mg/kg ip) followed by 1 h hypedocomotion test.Immediately after behavioral test,rats (n=4～6) were decapitated and tissues of nucleus accumbens (NAc) were isolated and prepared for neurotransmitters analysis by HPLC; Another twenty one rats were divided into control and TFF3-treatment group,and the rats were trained with a modified 2-day cocaine CPP conditioning procedure.During cocaine CPP conditioning process,saline or TFF3 (0.1 mg/kg ip) was injected 30 min prior to cocaine injection (5 mg/kg ip).Results Systemic administration of TFF3 (0.1 mg/kg,ip) significantly increased the cocaine-induced locomotor activity (Total distance in 1 hour were (180±41) cm,(359±53) cm,(590±75) cm,(153±27) cm for Control,Vehicle + Cocaine,TFF3+Cocaine and TFF3+Vehicle groups respectively) and augmented cocaine rewarding effects in CPP(Post-training CPP score were (98± 18) s,(187±24) s for Vehicle + Cocaine,TFF3+Cocaine groups respectively).TFF3 (0.1 mg/kg ip) administration increased the dopamine concentration in the NAc induced by cocaine injection ((0.65±0.1) ng/ml,(1.24±0.14) ng/ml,(1.75±0.23) ng/ml,(0.74±0.21) ng/ml for Control,Cocaine,TFF3 + Cocaine and TFF3 + Vehicle groups respectively).Conclusion TFF3 is involved in regulation of behavioral response to cocaine,which is associated with the increasing of dopamine in the NAc.

Objective To investigate the feasibility, accuracy and senaitivety of echo tracking (ET) technology on carotid elasticity in rabbits. Methods Thirty-nine healthy New Zealand rabbits were divided into 4 groups,which group 1,2,3 were experimental groups and group 4 was control group. Experimental groups were fed with high cholesterol diet,control group were fed with basic diet. The parameters of elasticity of carotid were measured with ET technology, including pressure-strain elasticity modulus(Ep), stiffness parameter(β), arterial complianee(AC), pulse wave velocity( PWV), augmentation index (AI) at the interval of 0,4,8,12 weeks of the experiment respectively. Blood-fat level of all rabbits were checked and the common carotid artery was examined pathologically,which were compared and analyzed then. Results With the prolong of the experiment time,the Ep and β raised, but the AC reduced. There were significant difference between experimental group 1 (4 weeks) and basement,which were same between experimental group 2 (8 weeks) and experimental group 1, between experimental group 3(12 weeks) and experimental group 2( P<0.05). PWV increased comparing with basement in the end of 8 weeks,which increased obviously in the end of 12 weeks,meanwhile was higher than in the end of 8 weeks,which there were significant differences ( P<0.01 ). AI had no change all the time. Conclusions ET technology is accurate and reliable in detecting early atherosclerosis. Probe maintenance tool can keep the steady of checking progress, sensitive of result and repeatability.

A porcine reproductive and respiratory syndrome virus (PRRSV) was obtained from clinic samples. Genes 5 and 6 encoding for the viral glycoprotein 5 and a membrane protein of the PRRSV designated as HH08 were amplified by reverse transcription-PCR. These sequences were compared with reference sequences derived from different geographical locations. The results indicated that the virus belongs to the North American type rather than European. Comparative analyses of the genetic diversity between the PRRSV isolate HH08 and other Chinese as well as foreign reference strains of PRRSV were discussed based on the sequence comparison and the topology of phylogenetic trees constructed in this study.
Animals ; Base Sequence ; China/epidemiology ; Gene Expression Regulation, Viral/physiology ; Genetic Variation ; Molecular Sequence Data ; *Phylogeny ; Porcine Reproductive and Respiratory Syndrome/epidemiology/virology ; Porcine respiratory and reproductive syndrome virus/*genetics/*metabolism ; Sequence Alignment ; Swine ; Viral Envelope Proteins/genetics/*metabolism ; Viral Matrix Proteins/genetics/*metabolism

Chloromethyl ether and diclomethyl ether are statutory substances that cause occupational lung cancer. From 2021 to 2022, the Department of Occupational Diseases of the Eighth People's Hospital of Hebei Province successively received 4 cases of lung cancer from a chemical company that required occupational disease diagnosis. All four patients had a clear occupational history of chloromethyl ether and diclomethyl ether for more than 1 year, diagnosis of primary small cell lung cancer supported by relevant histopathology, immunohistochemistry, and tumor markers. All the 4 patients were diagnosed as occupational lung cancer (chloromethyl ether, diclomethyl ether) .

Chloromethyl ether and diclomethyl ether are statutory substances that cause occupational lung cancer. From 2021 to 2022, the Department of Occupational Diseases of the Eighth People's Hospital of Hebei Province successively received 4 cases of lung cancer from a chemical company that required occupational disease diagnosis. All four patients had a clear occupational history of chloromethyl ether and diclomethyl ether for more than 1 year, diagnosis of primary small cell lung cancer supported by relevant histopathology, immunohistochemistry, and tumor markers. All the 4 patients were diagnosed as occupational lung cancer (chloromethyl ether, diclomethyl ether) .

The aim of the study is to obtain an efficient expression of recombinant ubiquitin-like specific protease 1 (Ulp1) by gene engineering. We cloned the Ulp1p, active fragment (403 aa-621 aa) of Ulp1, from Saccharomyces cerevisia, and subcloned into pGEX/Rosetta (DE3) to form an expression plasmid, pGEX-Ulp1p-His6. In order to enhance the solubility of GST-Ulp1p-His6, we purified the fusion protein GST-Ulp1p-His6 by either glutathione S-transferase agarose or Ni-NTA resin chromatography, the purity was up to 98%. We utilized the protein to cleave the SUMO fusions, and the specific activity of GST-Ulp1p-His6 was 1.375 x 10(4) U/mg. This study showed that the recombinant protein GST-Ulp1p-His6 displayed high specificity and activity.
Cloning, Molecular ; Cysteine Endopeptidases ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; biosynthesis ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; enzymology ; Solubility

Objective:To explore the classification and treatment strategy of femoral neck fracture combined with ipsilateral intertrochanteric fracture.Methods:A retrospective analysis was performed of the 44 patients who had been admitted to Department of Orthopedics, Liangxiang Teaching Hospital, Capital Medical University from March 2003 to March 2019 for femoral neck fracture combined with ipsilateral intertrochanteric fracture. They were 19 males and 25 females, aged from 37 to 93 years (average, 77.9 years). According to the anatomical location and displacement severity, the femoral neck fractures were divided into 3 types while the intertrochanteric fractures were classified as stable or unstable ones. There were 3 cases of type Ⅰ which were completely extracapsular ones, 31 cases of type Ⅱ which were intracapsular stable ones, and 10 cases of type Ⅲ which were intracapsular unstable ones. Types Ⅰ and Ⅱ fractures were treated with intramedullary fixation, and type Ⅲ fractures with cemented hip hemi-replacement+reduction and fixation of the intertrochanteric fracture with Kirschner wires and steel cables. Recorded were fracture healing time, function of the affected hip and complications.Results:The 44 patients were followed up for at least 2 years. The fracture healing time for the 3 patients with type I fracture averaged 5.6 months (from 4.4 to 6.8 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 2 cases and good in one case. For 30 of the 31 patients with type Ⅱ fracture, the fracture healing time averaged 7.2 months (from 5.1 to 9.3 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 18 cases, good in 6 cases, fair in 5 cases, and poor in 2 cases, giving an excellent and good rate of 77.4% (24/31). As for complications, withdrawal or cutting-out of the head screw happened in 6 cases, infection in one case and nonunion in one case. In 10 patients with type Ⅲ fracture, the hip function at the last follow-up was excellent in 7 cases, good in 2 cases and poor in one case.Conclusions:For type Ⅰ and type Ⅱ femoral neck fractures combined with ipsilateral intertrochanteric fracture, intramedullary fixation with angulation stability may be a proper choice. For type Ⅲ ones, hip joint replacement should be the first choice.

Objective To explore the safety and feasibility of laparoscopic hepatectomy combined with radiofrequency ablation for primary liver cancer difficult to manage.Methods A retrospective study was conducted to analyze the clinical data of 16 patients who underwent laparoscopic hepatectomy combined with radiofrequency ablation in the Zhejiang Provincial People's Hospital from Apr 2015 to Dec 2017.Results 2 more tumors were found by intraoperative laparoscopic ultrasound.All patients underwent laparoscopic hepatectomy combined with radiofrequency ablation successfully.There were 2 laparoscopic left hepatectomy combined with radiofrequency ablation,2 laparoscopic right hepatectomy combined with radiofrequency ablation,6 laparoscopic left lateral lobectomy combined with radiofrequency ablation,4 laparoscopic right postrior lobectomy combined with radiofrequency ablation and 2 irregular laparoscopic hepatectomy combined with radiofrequency ablation.The intraoperative blood loss ranged from 100-800 ml.The average operative time was (283 ± 112) min.The length of postoperative hospital stay ranged from 5 to 12 days.The tumor-free survival rate after operation is 100％ (16/16) on 6 to 38 months follow up.Conclusion It is safe and feasible to carry out laparoscopic hepatectomy combined with radiofrequency ablation in difficult to manage primary liver cancer.

Objective:To explore the clinical characteristics and genetic etiology for a Chinese pedigree affected with Dyschromatosis symmetrica hereditaria (DSH) in conjunct with developmental delay.Methods:A child who had presented at the First Affiliated Hospital of Zhengzhou University on May 28 2021 for abnormal skin pigmentation of the extremities and growth retardation for over 2 years was selected as the study subject. Clinical data of the child and his pedigree (11 individuals from three generations) was collected. The child was subjected to whole exome sequencing, and candidate variant was verified by Sanger sequencing.Results:The child, a two-year-and-seven-month-old male, had hyper- and hypopigmentation on his hands, feet and face, in addition with delayed development. All members of his pedigree had typical presentation of DSH. A heterozygous c. 2657G>A variant was found in exon 8 of the ADAR gene in the child, his mother, and elder sister. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted as likely pathogenic (PM1+ PM2_Supporting+ PP1+ PP3). Conclusion:The c. 2657G>A variant of the ADAR gene probably underlay the DSH in this pedigree.