To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective To explore the expression of 10 kDa interferon-gamma-induced protein(IP-10)in pathological scar and its role in the pathogenesis of pathological scar.Methods Imrnunohistochemistry was performed to detect the expression and distribution of IP-10 in 28 patients with keloid(group K),34 patients with hypertrophic scar(group HS),and 20 normal controls(group N).The data were collected and analyzed statistically.Results The expression of IP-10 was significantly higher in groups K and HS than in group N(P＜0.01),but no significant difference in the expression of IP-10 was found between groups K and HS.Conclusion IP-10 may enhance the formation of pathological scar by attracting T lymphocytes and inducing immune/inflammatory response.

Objective To analyze the influencing factors of quality of life of patients with occupational pneumoconiosis in Methods Chongqing using a structural equation model. A total of 1 181 pneumoconiosis patients in Chongqing were selected - as the research subjects using convenient sampling method. The Chinese version of the SF 36 Scale was used to investigate the quality of life of the patients. Amos 24.0 software was used to construct a structural equation model reflecting the quality of life of Results ， （ ），（ ） （ ）， patients. The scores of physical health mental health and quality of life were 61.7±18.5 68.7±20.5 and 64.7±16.8 respectively. The scores of the eight dimensions of the quality of life of the research subjects were lower than that of the （ P ） Chongqing City municipal and national norms all <0.01 . Multiple linear regression results showed that personal monthly ， ， ， income pneumoconiosis stage complications employment status and smoking were the influencing factors of the quality of life （ P ） ， of pneumoconiosis patients all <0.05 . The results of the structural equation model showed that the stage of pneumoconiosis ， ， complications personal monthly income and pulmonary rehabilitation treatment could directly affect the physical and mental （ P ） ， health of the research subjects all <0.05 . In addition the stage of pneumoconiosis and complications indirectly affected the （ P ）Conclusion physical and mental health of patients by affecting pulmonary rehabilitation treatment all <0.05 . The structural equation model can be used to verify the influencing factors of the quality of life of patients with occupational pneumoconiosis. ， Complications personal monthly income and pneumoconiosis stage are the main factors affecting the quality of life of patients with occupational pneumoconiosis.

OBJECTIVETo observe the effects of prenatal exposure to lead on nephroblastoma over-expressed gene (NOV) protein and mRNA expression in hippocampus of rats' offspring, and to explore the molecular mechanism of lead on learning and memory.

METHODSThe pregnant rats were divided into 1 control group and 3 lead expose groups randomly: low( 125 mg/L), middle (250 mg/L) and high (500 mg/L). 8 rats in each group. From pregnancy ld until birth, the rats were given double evaporated water or lead acetate water of different doses according to their groups. The samples of descendants were taken on embryo 18 th day, postnatal 1st day, 21st day, 60th day. The contents of lead in blood and hippocampus were determined by hydride generation atomic absorption spectrometry method. The expression of NOV protein and mRNA in hippocampus were observed by immunohistochemistry and in situ hybridization.

RESULTSThe lead contents of blood [(312.46 +/- 43.55), (419.35 +/- 62.25), (541.45 +/- 47.90) microg/L] and hippocampus[(2.10 +/- 0.18), (2.58 +/- 0.12), (3.41 +/- 0.23) microg/L] were significantly higher in lead exposed groups than that of control [(214.31 +/- 40.77), (0.76 +/- 0.13) microg/L] (P < 0.05) on the embryo 18th, 1st and 21 st day, while there was no significantly difference among them on 60 th day. The expression of NOV protein in all lead exposed groups were significantly decreased compared with control group (P < 0.05) on 1st and 21 st day, while there was no significantly difference among them on 60th day. The expression of NOV mRNA of all the lead exposed groups were significantly decreased compared with control group (P < 0.05) on the embryo 18th, 1st and 21st day, while there was significantly difference only in the high dose group (0.0355 +/- 0.0100) compared with control (0.0900 +/- 0.0200) (P < 0.01) on 60th day.

CONCLUSIONPregnancy low level lead exposure could decrease the NOV protein and mRNA expression in hippocampus of offspring, which might be one of the molecular mechanisms of effect of lead on learning and memory.

Animals ; Female ; Gene Expression ; Hippocampus ; metabolism ; Lead ; adverse effects ; blood ; Nephroblastoma Overexpressed Protein ; genetics ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVETo observe the change of nephron damaged by chemotherapy and to evaluate the effect of Baoshen Mixture (, BSM) in protecting and treating damaged nephrons.

METHODSFour hundred tumor patients with normal renal function and ready to receive chemotherapy were randomly assigned to two groups. Both groups received one cycle of chemotherapy program of 28-30 days with conventional hydratization, alkalization and chloridization. To the 200 cases in the treated group BSM was given orally thrice a day, 150 mL every time for 15 successive days and the other 200 cases in the control group were treated by chemotherapy alone. The clinical efficacy was compared after treatment, and the changed condition of damaged nephrons were monitored dynamically and compared at different time points (the 3rd, 7th, 14th and 21st day after chemotherapy) by measuring the micro-globulin beta(2) (beta(2)-MG), albumin (Alb) and immunoglobulin G (IgG) levels in urine with radioimmunoassay (RIA).

RESULTS(1) The effective rates in the treated group at the 4 time points of observation were all higher than those in the control group respectively (P<0.05 or P<0.01); (2) Less occurrence of abnormal beta(2)-M, Alb and IgG levels on the 14th and 21st day in the treated group took place compared to that in the control group (P<0.01); (3) Urinary levels of beta(2)-MG, Alb and IgG reached the peak on the 7th day in both groups, and then, they came down gradually and returned to the normal level on the 21st day. However, comparison between the two groups showed that all the three parameters in the treated group on day 3, 14 and 21 were lower than the respective one at the corresponding time points in the control group (P<0.05 or P<0.01).

CONCLUSIONThe chemotherapy damage on nephron is regular in time, and reversible when treated suitably. TCM shows a marked effect in protecting and treating the damage on nephron caused by chemotherapy.

Adult ; Aged ; Albuminuria ; prevention & control ; Antineoplastic Agents ; adverse effects ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin G ; urine ; Male ; Middle Aged ; Nephrons ; drug effects ; beta 2-Microglobulin ; urine

Abortion, Induced ; adverse effects ; Adult ; Device Removal ; Embolization, Therapeutic ; Female ; Humans ; Intrauterine Devices ; Pregnancy ; Uterine Artery ; Uterine Rupture ; etiology

OBJECTIVETo observe the the expression of endoplasmic reticulum stress (ERS) related factors in deep tissue injury (DTI) at pressure ulcer rat and to investigate the ERS mechanism of DTI in muscle tissue and protective effect of 4-phenylbutyric acid (4-PBA) in local tissue.

METHODSFifty male SD rats were randomly devided into control group, model group, experimental group NS group and PBA group, the experimental groups were divided into 4 d, 7 d, 14 d and 21 d group according to the observation time (n = 5). Rats in the PBA group were administrated with gastric perfusion of 4-PBA after the modeling; the NS group was given normal saline of the same quantity. Using HE staining to observe morphologic character. The expression of glucose regulated protein 78 (GRP78), CHOP, Caspase 12 were detected by immunohistochernical staining. Cell apoptosis was detected by TUNEL assay.

RESULTSHE staining results showed that each group demonstrated compression injury compared with control group: cellular swelling, ompaction of nuclear, and apoptosis in muscle tissue. The new muscle fiber in 4-PBA group fused faster than those in NS group. The number of TUNEL positive cells peaked at 4 day after compression, then got decreased on day 7 in muscle tissue, apoptosis positive cells were diminished after 4-PBA treatment. The immunohistochemical staining results showed that the expression of protein GRP78, CHOP, Caspase 12 peakd 4 d after modeling and decreased gradually. The GRP78, CHOP, Caspase 12 protein expression were significantly higher than those of PBA group at all time points (P < 0.05).

CONCLUSIONCell apoptosis induced by endoplasmic reticulum stress took part in deep tissue injury resulting of pressure ulcer, which mechanism might be related to reducing apoptosis mediated by CHOP, Caspase 12.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Male ; Muscle, Skeletal ; pathology ; Phenylbutyrates ; pharmacology ; Pressure Ulcer ; physiopathology ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Transcription Factor CHOP ; metabolism

Adolescent ; Child ; Child, Preschool ; Complement System Proteins ; analysis ; Female ; Humans ; Immunoglobulins ; analysis ; Infant ; Male ; Pneumonia, Mycoplasma ; immunology

A simple and sensitive assay for rapid detection of human coronavirus NL63 (HCoV-NL63) was developed by colorimetic reverse transcription loop-mediated isothermal amplification (RT-LAMP). The method employed six specially designed primers that recognized eight distinct regions of the HCoV-NL63 nucleocapsid protein gene for amplification of target sequences under isothermal conditions at 63 degrees C for 1 h Amplification of RT-LAMP was monitored by addition of calcein before amplification. A positive reaction was confirmed by change from light-brown to yellow-green under visual detection. Specificity of the RT-LAMP assay was validated by cross-reaction with different human coronaviruses, norovirus, influenza A virus, and influenza B virus. Sensitivity was evaluated by serial dilution of HCoV-NL63 RNA from 1.6 x 10(9) to 1.6 x 10(1) per reaction. The RT-LAMP assay could achieve 1,600 RNA copies per reaction with high specificity. Hence, our colorimetric RT-LAMP assay could be used for rapid detection of human coronavirus NL63.
Colorimetry ; methods ; Coronavirus Infections ; diagnosis ; virology ; Coronavirus NL63, Human ; genetics ; isolation & purification ; DNA Primers ; genetics ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcription ; Sensitivity and Specificity

Neuropathic pain is a chronic pain caused by primary nervous system damage and nerve dysfunction. Its pathogenesis is complex and diverse. It is difficult to treat clinically. In recent years, researchers used electroacupuncture to treat neuropathic pain and obtained a desirable effect. This article summarizes recent years’ studies on the main mechanisms of neuropathic pain and the intervention effect of electroacupuncture to provide reference for following studies on electroacupuncture treatment of neuropathic pain.