In order to develop ELISA method for measuring the degradation fragment of fibronectin (MAD2),hybridoma technique was used to obtain the monoclonal antibody (McAb)IgG 1 against MAD2 without cross reaction with fibronectin.The sera from 277 patients with hepatocellular carcinoma (HCC),76 with metastatic hepatic carcinoma (HMC),98 with alimentary canal carcinoma (ACC) and 156 with chronic liver disease(CLD) and 48 healthy subjects were assayed with ELISA method using this antibody.The examination showed that the mean value of MAD2 from the patients with HCC showed obviously significant difference compared with those of CLD,HMC,ACC and normal control groups ( p

Pre-S2 protein in 78 samples of liver from patients with various types of hepatitis B was determined by immunohistochemical method.45 out of 78 samples (57.69%) were positive for pre-S2.The location and expression patterns of pre-S2 were similar to those of HBsAg,i.e.,pre-S2 was found in cytoplasm of hepatocytes,epithelial cells of bile ducts,and hepatocellular carcinoma cells,etc,Four different patterns diffuse,inclusion,submembranous,and membranous type were demonstrated.The membranous expression of pre-S2 was often associated with activity of liver diseases.The positive rate of pre-S2 was significantly higher in HBcAg positive group than HBcAg negative group.It seems that pre-S2 protein of HBV may be a replicative marker of HBV.

Objective To clearify the features of an outbreak of Guillain-Barré syndrome (GBS) from Shuangyang district of Changchun city, Jilin province, China.Methods Serum samples obtained were tested serologically for antibodies to chlamydia, coxsackie virus, cytomegalovirus, and hepatitis B virus.Stool samples obtained were cultivated to search Campylobacterjejuni.Electrudiagnostic studies were done 1 day after onset to 10 days.Results Twenty-two patients (12 males and 10 females, aged 15-71 years) were from Shuangyang district, a suburb of Changchun, during a 2-week period in June to July 2007.An illness in the preceding 4 weeks was reported in 17 patients, 16 gave a history of diarrhoea, 1 patient had symptoms consistent with an "upper respiratory infection" and 1 patient had both.Limb weakness were the earliest symptoms.Respiratory assistance was needed by 3 patients.Tendon reflexes were lost as weakness developed.Eleven patients had raised concentrations of protein in the cerebrospinal fluid ((0.74±0.32) g/L), and the averaged cell count was 3×106/L There were 3 kinds of antibodies of hepatitis B virus positive above 17 GBS patients.Electrodiagnostic studies showed severe reductions in motor evoked amplitudes from distal stimulation in 12 patients.Sensory action potentials were normal.Conclusions The distinctive epidemiologicai, clinical and electromyography characteristics of these patients suggest that the disorder, different from acute inflammatory demyelinating polyneuropathy, is concordant with acute motor axonal neurupathy.The features of Shuangyang GBS was similar to those previously reported in several aspects.

To express recombinant Ancylostoma caninum anticoagulant peptide-c2 (AcAPc2), a whole cDNA fragment encoding AcAPc2 was achieved by ligation- PCR and inserted into prokaryotic expression vector pTWIN1 for constructing the specific self-splicing prokaryotic expression vector, pTWIN1-AcAPc2; positive recombinants were transformed into E. coli ER2566 for expression research. The recombinant protein, AcAPc2-intein2-CBD, was soluble and expressed in E. coli ER2566 (about 30.1% fusion protein in total protein). AcAPc2-intein2-CBD was characterized to be 41 KD by SDS-PAGE and identified by Western-blot. The recombinant fusion protein was purified to a efficiently high degree by chitin affinity chromatography. After the process of specific self-splicing induced by beta-Mercaptoethanol, the target protein, AcAPc2, was obtained, characterized to be 21 KD by SDS-PAGE and migrated as a dimmer. Molecular weight of AcAPc2 conformed to native dimmer. Bio-information analysis indicated relationship between secondary construction of AcAPc2 and biologic function. These findings greatly facilitate the purification of AcAPc2 and are very important for the additional studies on its anti-coagulation mechanism and its clinical application as anti-coagulation medicine.
Animals ; Dogs ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genes, Helminth ; Genetic Vectors ; Helminth Proteins ; biosynthesis ; genetics ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; RNA Splicing ; Recombinant Fusion Proteins ; chemistry ; pharmacology

BACKGROUNDTo express recombinant non-structural protein 3 of hepatitis C virus (HCV) in E. coli.

METHODSThe non-structural 3 (NS3) region DNA fragment of HCV was amplified by polymerase chain reaction (PCR) and inserted into inducible proeukaryotic expressive vector pET 30C(+)at Bam H1/EcoR1 sites. The competent BL21 (DE3) E.coli was transformed, and then cultured and induced with IPTG. The expressed HCV NS3 protein was confirmed with ELISA and dot blot hybridization using HCV NS3-specific single chain Fv (ScFv) antibody.

RESULTS1 893 bp DNA fragment of HCV NS3 coding region was amplified by PCR technique. HCV NS3 expressive vector pET-NS3 was constructed. After transformation with pET-NS3 and induction with IPTG, recombinant HCV NS3 protein was expressed and confirmed by specific ELISA and dot blot hybridization.

CONCLUSIONSThe recombinant HCV NS3 can be expressed in E. coli.

Escherichia coli ; genetics ; Gene Expression ; Hepatitis C Antibodies ; biosynthesis ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.