Recombinant human interferon α2b(rhIFNα2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNα2b is complex.In this study,an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b.RhIFNα2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNα2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other com-mercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.

Objective To investigate the expression of unfolded protein response (UPR) gene glucose regulated protein 78(GRP78) and X-box binding protein 1(XBP1) in esophageal squamous cell carcinoma, its effect on activation of the endoplasmic reticulum stress (ERS) and its mechanism in the growth of esophageal squamous cell carcinoma. Methods The expressions of GRP78 and XBP1 were detected by immunohistochemistry in 30 samples of esophageal squamous cell carcinoma and 30 samples of normal esophageal squamous epithelium. The correlation between expressions of both proteins and prognosis was analyzed. Results GRP78 positive rate was 83.3 %(25/30) in esophageal carcinoma, while the proportion was 20.0 %(6/30) in normal esophageal (χ2=25.833, P<0.05). XBP1 positive rate was 70.0 % (21/30) in esophageal carcinoma, while the proportion was 26.7%(8/30) in normal esophageal(χ2=20.872, P<0.05). The positive rates of GRP78 and XBP1 in invasive muscular layer of esophageal squamous cell carcinoma were significantly higher than those in invasive mucous layer. Conclusion GRP78 and XBP1 are highly expressed in esophageal squamous cell carcinoma, which may involve the occurrence and development of the esophageal carcinoma.

OBJECTIVETo investigate the protective effect of sesamin against cadmium chloride (CdCl2)-induced cardiotoxicity in rats.

METHODSFifty male Wistar rats were randomly assigned to five groups: control group, CdCl2 group, and low-, middle-, and high-dose sesamin groups. The control group was given normal saline. The CdCl2 group and sesamin groups were intraperitoneally injected with CdCl2 (5 mg/kg×2 d), and the low-, middle-, and high-dose sesamin groups were given 20, 40, and 80 mg/kg sesamin, respectively. All treatments lasted for four weeks. ECG was measured by a physiological recorder, and serum myocardial enzyme levels were determined by biochemical assay. The heart was weighed, and heart tissues were used in histopathological examination and determination of malondialdehyde (MDA) level.

RESULTSCompared with the control group, the CdCl2 group showed significantly higher levels of serum CK and CK-MB, an increased heart coefficient, significant ST-segment elevation, and higher level of MDA in myocardial tissue (P < 0.05). Histopathological analysis showed edema of myocardial tissues and cells, myocardial fibers disorder, karyopyknosis, and uneven or deep staining of nuclear chromatin. Different doses of sesamin relieved the myocardial pathological changes induced by CdCl2, and high-dose sesamin was the most effective. The middle- and high-dose sesamin groups showed significantly reduced serum CK and CK-MB levels compared with the CdCl2 group (P < 0.05). The heart coefficient of the high-dose sesamin group (0.19±0.01%) was significantly lower than that of the CdCl2 group (0.21±0.01%) (P < 0.05). Myocardial MDA levels of the three sesamin groups (42.32±4.65, 36.71±5.34, and 33.12±4.62 nmol/mg pro, respectively) were all significantly lower than that of the CdCl2 group (55.87±3.65 nmol/mg pro) (P < 0.05).

CONCLUSIONSesamin can relieve myocardial injury induced by CdCl2, and one possible mechanism is the enhancement of antioxidant capacity of myocardial tissue.

Animals ; Cadmium Chloride ; toxicity ; Creatine Kinase, MB Form ; blood ; Dioxoles ; pharmacology ; Heart ; drug effects ; Lignans ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar

OBJECTIVE To observe the protection of vitamin C on the cardiac injury induced by 50 nm titanium dioxide inmice.METHODS Kunming mice were ad mistered by ig of vitamin C 100,200 and 400 mg·kg -1 for 2 d.And then the mice were ad mistered by ig of nano-TiO2 2 g·kg -1 and vitamin C (100.0,200.0 and 400.0 mg·kg -1 )for 3 d,the interval of treatment with nano-TiO2 and vitamin C was 4 h.The mice were scarified 24 h later after the last ad ministration.Electrocardiogra m (ECG)was determinated by physiological recorder.The myocardial enzy mes activities in serum and superoxide dismutase (SOD)and glutathione peroxidase(GSH-Px)activities in serum and myocardial tissue were determinated by bioche mical method.Cometassay was used to detect the DNA da mage of the heart. Heart tissue was used for histopathological exa mination by HE staining.RESULTS Co mpared with the control,ECG showed higher S-T and T-wave a mplitude of nano-TiO2 2 g·kg -1 (P<0.05).The myocar-dial enzy mes activities significantly increased and activities of SOD and GSH-Px significantly decreased in nano-TiO2 group,compared with the control group(P <0.05).Cometassay showed that olive tail mo ment (OTM)was significantly increased after nano-TiO2 2 g·kg -1 ,compared with the control group (P<0.05).The histopathology showed ede ma of myocardial cells,myofibril disorders and increasing infla mmatory cells.Vita min C 100,200 and 400 mg·kg -1 can decrease S-T in ECG,OTM,myocardial enzy mes activities,increase the SOD and GSH-Px activities in serum and myocardial tissue;reduce myocardial hypertrophy and infla mmatory cells.CONCLUSION nano-TiO2 can induce myocardial injury inmice and vitamin C can alleviate the da mage.The mechanism may be associated with the antioxidant ability of vitamin C inmyocardial tissue.

Objective To analyze the influence of the cesarean section and vaginal delivery at term pregnancy when fetal distress in labor conditions on maternal and neonatal outcomes .Methods 238 cases of fetal distress in labor when full-term pregnant were selected ,and they were randomly divided into the observation group and control group,119 cases in each group .The control group was given midwifery vaginal delivery ,while the observation group was given cesarean section .The maternal and neonatal outcomes were compared between the two groups . Results 78 cases(65.55%) of neonatal score above 7 or more points and 7 or more points of the control group were 58 cases(48.74%),the difference was significant (P<0.05).The incidence rate of neonatal complications in the observation group was 5.88%, which was significantly lower than 20.16% of the control group ( P <0.05 ). Conclusion Fetal distress in full-term pregnancy in labor , the use of cesarean section has higher neonatal score compared with vaginal delivery ,and it has low incidence rate of neonatal complications ,it is worth to be applied in clinical.

This paper reports the determination of the drug antibody ratio in an antibody-drug conjugate with two methods, i.e. LC-MS and UV/VIS, and to provide a reliable method to scientifically evaluate and effectively control the drug antibody ratio. Deglycosylated sample was analyzed with C4 column followed by MS, and the number of conjugated drugs in the antibody was determined by the molecular weight increase due to the addition of different number of drugs to the antibody, and then drug antibody ratio was calculated by weighted average of different number of drugs conjugated to the antibody. Optical density at 252 and 280 nm was measured with UV/VIS, and due to the difference of extinction coefficients between the antibody and the drug, the drug antibody ratio was calculated from linear equation with two unknowns. The drug antibody ratio was 3.21 and 3.25 respectively measured by the two methods, and the results were similar with the two methods. Our study indicated that both methods, LC-MS and UV/VIS, could be applied to the analysis of drug antibody ratio of the antibody drug conjugate.

The hydrophilicity of the normal decoction pieces (NDP) of Indigo Naturalis is not good, therefore, it is not suit for decoctions. In this paper, powder modification technology is used and some NDP and alcohol are ground together in the vibromill to prepare the hydrophilic decoction pieces (HDP) of Indigo Naturalis. Initially, the properties of NDP, ultrafine decoction pieces (UDP) and HDP are compared, the hydrophilicity of UDP was promoted slightly, that of HDP is promoted dramatically. Then, three batches of Indigo Naturalis are prepared to HDP separately, but there is no obvious difference in the contact angle. Furthermore, the size distribution, surface area and micro-shape of HDP are bigger than that of UDP and smaller than NDP. The contents of indigo and indirubin in three decoction pieces are the same, as well as the species of inorganic substance, although there is a little difference in the proportion of five inorganic substances. The fact suggests the change of physical state and the qualitative and quantitative change of organism and inorganic substances are not the main factors to influence the hydrophilicity. In addition, hydroxyl, methylene and methyl can be identified at the wavenumber of 3 356 cm(-1) and 1 461 cm(-1) in infrared spectrum; the content of alcohol in HDP is 0.67% measured by gas chromatogram. The stability of HDP in the heating condition is studied, the fact suggests the hydrophilic effect of HDP at 40 degrees C is relatively stable. All above research suggests that the alcohol is the main factor to influence the hydrophilicity and maybe the intermolecular force which fixed alcohol molecule on the surface of Indigo Naturalis is the basic principle to produce the hydrophilicity.

The primary structure of TNK-tissue plasminogen activator (TNK-tPA) was characterized using liquid chromatography-mass spectrometry (LC-MS). Firstly, the molecular mass of deglycosylated protein was measured. Then peptide mass mapping and MS/MS of the reduced, alkylated and trypsin-digested sample were tested and analyzed so as to verify its amino acid sequence and identify post-translational modifications. Results show that the amino acid sequence was consistent with designed structure; about 5% of M207 was oxidized; T61 was fucosylated with -80% occupancy; N103, N448 and N184 (-15% occupancy) were glycosylated with complex-type oligosaccharides. LC-MS coupled with proper sample pretreatment is approved to be a rapid and powerful approach to characterize the primary structure of TNK-tPA.

The amino acid sequence of the fusion protein FP3 was measured by two types of LC-MS/MS and its primary structure was confirmed. After reduction and alkylation, the protein was digested with trypsin and glycosyl groups in glycopeptide were removed by PNGase F. The mixed peptides were separated by LC, then Q-TOF and Ion trap tandem mass spectrometry were used to measure b, y fragment ions of each peptide to analyze the amino acid sequence of fusion protein FP3. Seventy-six percent of full amino acid sequence of the fusion protein FP3 was measured by LC-ESI-Q-TOF with the remaining 24% completed by LC-ESI-Trap. As LC-MS and tandem mass spectrometry are rapid, sensitive, accurate to measure the protein amino acid sequence, they are important approach to structure analysis and identification of recombinant protein.

The digital library refers to the delivery of networked book and information service which has sprung up with the development of the Internet and which has been realized by means of computer technology. The authors offer a description of the fundamental form, structure and model of the digital library, and expound relevant standards, functions of the chief software, technical indexes and ways of realization through actual practice in building the system architecture of a digital hospital library.