In order to screen African swine fever virus (ASFV) diagnostic antigen with the best enzyme linked immunosorbent assay (ELISA) reactivity. By establishing the ELISA method, the diagnostic antigen of ASFV p30 protein expressed by baculovirus-insect cell expression system as reference, we explored the antigenic properties and diagnostic potential of ASFV p35 protein expressed by prokaryotic expression system as a diagnostic antigen. The results of Western blotting and immunofluorescence show that the molecular weight of the recombinant p35 protein and p30 protein obtained was 40 kDa and 30 kDa, respectively, and these two proteins had good immuno-reactivity with ASFV positive serum. Recombinant p30 and p35 proteins were used as diagnostic antigens to establish ELISA, and the sensitivity and repeatability of these methods were tested. The results show that although the detection sensitivity of the p30-ELISA established in this study was higher than that of the p35-ELISA, the sensitivity of p35-ELISA was 95.8%, and variations in intra- and inter-assay repeatability of the two methods were less than 10%. The coincidence rate between the p35-ELISA and the imported kit was 97.2%. Results show that p35-ELISA was sensitive and stable, and could detect specific antibodies against ASFV.
African Swine Fever/diagnosis* ; African Swine Fever Virus/genetics* ; Animals ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Recombinant Proteins/genetics* ; Swine

Objective:To explore the effect of cognitive-behavioral therapy on psychological stress and quality of life in patients with pulmonary tuberculosis.Methods:According to two-level cluster random design 461 patients with tuberculosis from 20 communities in Pizhou county of Jiangsu province were selected in the study from September 2018 to November 2018. The intervention group received cognitive-behavioral therapy for two months, while control group received routine follow-up. Anxiety, depression and quality of life were assessed by GAD-7, PHQ-9 and SF-36 scales, respectively. At the same time, the comparison between the two groups was conducted by independent sample t test, and the difference between the two groups before and after treatment was analyzed by paired sample ttest. Results:A total of 454 participants were finally included in this analysis; there were 230 cases in the intervention group and 224 cases in the control group. In the intervention group the scores of anxiety and depression after intervention were significantly lower than the baseline scores [(7.57±5.27) vs. (5.93±2.56), t=-4.245, P<0.01; (8.13±6.01) vs. (6.02±2.67); t=-4.866, P<0.01], and the quality of life score was significantly higher than the baseline score [(58.46±12.71) vs. (74.31±13.22); t=13.108, P<0.01]; while in the control group there were no significant differences in the scores of anxiety, depression and quality of life after intervention, compared with those at baseline [(7.62±5.41) vs.(7.65±5.38); (8.00±5.84) vs. (8.07±5.91); (59.11±13.25) vs. (60.51±13.76); t=0.059, t=0.126, t=1.104, all P>0.05]. However, only for patients with mild and moderate anxiety and depression symptoms in the intervention group, the anxiety and depression scores were decreased after intervention [(7.29±1.21) vs. (5.54±1.71), (11.99±1.31) vs. (9.17±1.55); (7.01±1.47) vs. (4.42±1.22), (11.88±1.12) vs. (8.39±2.33); t=8.056, t=10.020, t=13.558, t=8.852,all P<0.01]. Conclusion:Cognitive-behavioral therapy can relieve the psychological pressure and improve the quality of life in pulmonary tuberculosis patients with mild or moderate anxiety/depression symptoms.

To screen the best genotypeⅠJapanese encephalitis virus subunit vaccine candidate antigens, the prMEIII gene, the polytope gene and the prMEIII-polytope fusion gene of the GenotypeⅠJapanese encephalitis virus GS strain were cloned into prokaryotic expression vector pET-30a. The recombinant proteins were obtained after the induction and purification. The prepared recombinant proteins were immunized to mice, and the immunogenicity of the subunit vaccine candidate antigens was evaluated through monitoring the humoral immune response by ELISA, detecting the neutralizing antibody titer by plaque reduction neutralization test, and testing the cell-mediated immune response by lymphocyte proliferation assay and cytokine profiling. The recombinant proteins with the molecular weights of 35 (prMEIII), 28 (polytope antigen) and 57 kDa (prMEIII-polytope) induced strong humoral and cellular immune responses in mice. Compared with prMEIII-polytope and polytope proteins, the prMEIII protein induced a significant expression of IL-2 and IFN-γ (P<0.05) and the significant lymphoproliferation of splenocytes (P<0.05). The neutralizing antibody titer induced by the prMEIII protein was close to that induced by the commercial attenuated vaccine SA14-14-2 (P>0.05). The study suggests that the prMEIII protein can be used for the development of the Japanese encephalitis virus subunit vaccine.
Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; immunology ; Encephalitis Virus, Japanese ; immunology ; Encephalitis, Japanese ; immunology ; prevention & control ; Immunogenicity, Vaccine ; Mice ; Mice, Inbred BALB C ; Vaccines, Subunit ; immunology ; Viral Vaccines ; immunology

Objective To explore the value of 3D time-of-flight MRI ( MRI 3D-TOF) combined with 3D reconstruction technique for the diagnosis and treatment of vascular compression of trigeminal neuralgia by magnetic resonance angiography combined with three-dimen-sional reconstruction technique.Methods A total of 80 patients with vascular compression of the trigeminal neuralgia in our hospital from January 2012 to February 2014 were diagnosed by axial MRI 3D-TOF,combined with 3D vascular and nerve reconstruction in the anatomy Department for further diagnosis and treatment.Results The patients were diagnosed according to the trigeminal nerve and the typings of pe-ripheral vascular compression.The diagnosis results for 29 cases of typeⅠ,19 cases of typeⅡ,32 cases of typeⅢfrom physician 1,the di-agnosis results for 48 cases of typeⅠ,21 cases of typeⅡand typeⅢin 11 cases from physician 2,the difference of the diagnosis bwtween two physicians was statistical significance (P=0.00).Compared with intraoperative diagnosis,the diagnosis of 3D reconstruction results was similar to the intraoperative diagnosis,and the difference was no statistical significance (P=0.98).Conclusion MRI 3D-TOF combined with three-dimensional reconstruction technique in the diagnosis of vascular trigeminal neuralgia shows clear images of the nerve and its sur-rounding vessels,which can be applied to the diagnosis of trigeminal neuralgia.

OBJECTIVETo assess the clinical effects of combined endoscopic-laparoscopic technique for one-stage treatment of cholelithiasis with concomitant choledocholithiasis.

METHODSA retrospective analysis was conducted of the clinical data of 30 patients (Group A) with cholelithiasis and choledocholithiasis receiving one-stage laparoscopic cholecystectomy (LC) combined with intraoperative encoscopic retrograde cholangio-pancreatography (ERCP) and 32 patients (Group B) receiving LC combined with 1aparoscopic common bile duct exploration. The operative time, blood loss, conversion to open surgery rate, time to postoperative ambulation, calculi residual rate, hospitalization cost and length of hospital stay were analyzed comparatively.

RESULTSThere were statistically differences between the two groups in hospitalization cost and length of hospital stay (P<0.05) but not in the other indices (P>0.05).

CONCLUSIONCombined endoscopic-laparoscopic techniques can be a safe and feasible option for one-stage treatment of concomitant cholelithiasis and choledocholithiasis to allow rapid postoperative recovery with a shortened hospital stay.

Adult ; Aged ; Cholangiopancreatography, Endoscopic Retrograde ; methods ; Cholecystectomy, Laparoscopic ; methods ; Choledocholithiasis ; complications ; surgery ; Cholelithiasis ; complications ; surgery ; Combined Modality Therapy ; Female ; Humans ; Laparoscopy ; methods ; Length of Stay ; economics ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies

OBJECTIVETo screen tumor biomarkers in the plasma close related with hypopharyngeal carcinoma.

METHODSPooled plasma from 6 patients with hypopharyngeal carcinoma and 6 healthy individuals were collected. After removal of high-abundance plasma proteins, two-dimensional gel electrophoresis (2-DE) was performed to isolate the total proteins, and the protein spots with more than 2-fold differential expressions were detected by 2D analysis software followed by identification by MALDI-TOF/TOF mass spectrometer. Western blotting was performed to validate the expression level of α2-HS-glycoprotein.

RESULTSA total of 11 differentially expressed protein spots were selected, including 5 upregulated proteins and 6 downregulated proteins. MALDI-TOF/TOF identified the upregulated proteins in hypopharyngeal carcinoma patients as alpha-2-HS-glycoprotein and haptoglobin and downregulated ones as Ig kappa chain C region and apolipoprotein A-I. Western blotting demonstrated that α-2-HS- glycoprotein expression level was consistent with that detected by 2-DE.

CONCLUSIONPatients with hypopharyngeal carcinoma show different plasma protein profiles from healthy individuals. These differentially expressed proteins may serve as potential specific tumor biomarkers for hypopharyngeal carcinoma.

Biomarkers, Tumor ; blood ; Blood Proteins ; metabolism ; Humans ; Hypopharyngeal Neoplasms ; blood ; Male ; Middle Aged ; Proteomics ; methods

OBJECTIVE: To compare and analyze the expression difference of proteins between laryngocarcinoma and healthy individuals to search for protein biomarkers that may be detected in plasm of laryngocarcinoma patients. METHOD: Pooled plasm from 6 laryngocarcinoma patients and 6 healthy individuals as controls were collected. Two-dimensional gel electrophoresis (2-DE) was used to isolate the total proteins, and the differential protein spots were identified by MALDI-TOF/TOF mass spectrometer, and then the biological information of the proteins was analyzed. RESULT: Twenty differential expressed protein spots with more than 1.5-fold between the laryngocarcinoma and healthy individuals were selected, and there were 8 proteins upregulated and 12 proteins downregulated among these proteins. After identifying by MALDI-TOF/TOF, compared with healthy controls, the spots that were L1, C-reactive protein, haptoglobin, ceruloplasmin; the spots that were downregulated were: Serotransferrin, alpha-2-HS-glycoprotein, fibrinogen gamma chain, haptoglobin related protein, Ig lambda-1 chain C regions, Ig kappa chain C region, apolipoprotein A-I, transthyretin, apolipoprotein C-III. CONCLUSION Compared with laryngocarcinoma and healthy individuals, the plasm proteins profile showed differently. The proteins of differential expressed are expected to be the specific plasm biomarkers for laryngocarcinoma patients.
Biomarkers, Tumor ; blood ; Case-Control Studies ; Humans ; Laryngeal Neoplasms ; blood ; Male ; Mass Spectrometry ; Middle Aged ; Proteome ; metabolism ; Proteomics ; methods

Objective To assess the clinical effects of combined endoscopic-laparoscopic technique for one-stage treatment of cholelithiasis with concomitant choledocholithiasis. Methods A retrospective analysis was conducted of the clinical data of 30 patients (Group A) with cholelithiasis and choledocholithiasis receiving one- stage laparoscopic cholecystectomy (LC) combined with intraoperative encoscopic retrograde cholangio-pancreatography (ERCP) and 32 patients (Group B) receiving LC combined with 1aparoscopic common bile duct exploration. The operative time, blood loss, conversion to open surgery rate, time to postoperative ambulation, calculi residual rate, hospitalization cost and length of hospital stay were analyzed comparatively. Results There were statistically differences between the two groups in hospitalization cost and length of hospital stay (P<0.05) but not in the other indices (P>0.05). Conclusion Combined endoscopic-laparoscopic techniques can be a safe and feasible option for one- stage treatment of concomitant cholelithiasis and choledocholithiasis to allow rapid postoperative recovery with a shortened hospital stay.

Objective To assess the clinical effects of combined endoscopic-laparoscopic technique for one-stage treatment of cholelithiasis with concomitant choledocholithiasis. Methods A retrospective analysis was conducted of the clinical data of 30 patients (Group A) with cholelithiasis and choledocholithiasis receiving one- stage laparoscopic cholecystectomy (LC) combined with intraoperative encoscopic retrograde cholangio-pancreatography (ERCP) and 32 patients (Group B) receiving LC combined with 1aparoscopic common bile duct exploration. The operative time, blood loss, conversion to open surgery rate, time to postoperative ambulation, calculi residual rate, hospitalization cost and length of hospital stay were analyzed comparatively. Results There were statistically differences between the two groups in hospitalization cost and length of hospital stay (P<0.05) but not in the other indices (P>0.05). Conclusion Combined endoscopic-laparoscopic techniques can be a safe and feasible option for one- stage treatment of concomitant cholelithiasis and choledocholithiasis to allow rapid postoperative recovery with a shortened hospital stay.

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. Phosphotyrosine (pTyr) is the substrate for PTP1B dephosphorylation. Malonic acid moiety was used herein as a mimic of the phosphate group in pTyr, and novel malonic acid derivatives 1-7 were designed, synthesized and evaluated as PTP1B inhibitors. Results from enzymatic assays indicated that compounds 3 and 4 exhibited potent inhibition against human recombinant PTP1B with IC50 values of 7.66 and 1.88 micromol x L(-1), respectively.