MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.

Four compounds have been isolated from Sargassum fusiforme(HaTv. ) Setch. collected in the south Zhejiang province of China. Among these compounds the structures of three have been deduced as mannitol, fucosterol and saringosterol on the basis of chemical and IR,NMR,MS spectral data.

Objective To observe the apoptosis and expression of Toll-like receptor (TLR)-4's mRNA in ischemia-reperfusion injured rat renal tubular epithelia cells (NRK-52E) in vitro, and effect of Cordyceps Sinensis (CS) extractant.Methods Cultured NRK-52E cells were divided into a control group, a model group, and a CS preincubated group. All first removed media and incubated with antimycin A for 1 h, and then recovered the media to simulate the ischemia-reperfusion injury in vitro. We detected the apoptosis ratio of cells by flow cytometer and the mRNA expression of TLR-4, Bax, Bcl-2 gene by reverse transcription-polymerase chain reaction at different time points.Results In ischemia-reperfusion injured NRK-52E, the apoptosis ratio rose as time passed (P＜0.05). We also observed increased mRNA expression of TLR-4, Bax (P＜0.05) and deceased expression of Bcl-2 (P＜0.05). Compared with the model grpup, the CS preincubated NRK-52E cells showed apparent tolerance to ischemia-reperfusion injury, which manifestated lower apoptosis ratio (P＜0.05), decreased expression of mRNA of TLR-4, Bax and increased expression of Bcl-2 (All Ps＜0.05). Conclusion The number of apoptosis cells and the expression of TLR-4 mRNA inceased with ischemia-reperfusion injury of NRK-52E in vitro. CS can prevent the NRK-52E cells from ischemia-reperfusion injury by downregulating TLR-4 gene.

Aim To explore the inhibitory effect of dexamethasone on the expression of SH2-B? in the lung and the visceral sensory afferent system(C7-T5 spinal ganglia and the corresponding posterior horn of the spinal cord)of asthmatic mice.Methods Murine model of asthma of BABL/c mice was induced by ovalbumin in vivo.By means of immunohistochemistry and Western blot,the expression of SH2-B? in the lung,C7-T5 spinal ganglia and corresponding spinal cord was detected and the inhibitory effect of dexamethasone on it was observed.Inspiratory airway resistance was measured with AniRes 2003 lung function system.Results The expression of SH2-B? in the lung,C7-T5 spinal ganglia and the corresponding spinal cord of the asthmatic mice was much higher than that of the control group and the dexamethasone group(P

A type of water-soluble carboxymethyl chitosan (O--CMC), with 76% degree of substitution determined by conductivity method, was prepared by using chloroacetic acid to react with C6-OH of chitosan. The solubility of O--CMC was characterized also. Animal experiment in rabbits showed that O--CMC could lubricate arthron, inhibit proliferation of fibroblast cells on rabbits' knee joints, benefit the process of repairing pathologic articular cartilage, and produce good therapeutic effect on rheumatoid arthritis.
Animals ; Arthritis, Experimental ; drug therapy ; pathology ; Cartilage, Articular ; drug effects ; Cell Proliferation ; drug effects ; Chitosan ; administration & dosage ; analogs & derivatives ; therapeutic use ; Female ; Fibroblasts ; drug effects ; Injections, Intra-Articular ; Knee Joint ; drug effects ; Male ; Rabbits

Objective To explore the value of transthoracic echocardiography (TTE) and transesophageal echocardiography (TEE) in diagnosing adult multiple atrial septal defect (MASD). Methods Thirty adult patients with MASD were examined with TTE, 25 patients were examined also with TEE, 26 patients were examined with cardiac catheterization as well. Transcatheter closure of MASD was performed in 20 patients and succeeded in 18, while open-chest operation was performed in 4 patients. Results Foramen secundum atrial septal defect was diagnosed with both TTE and TEE with an accuracy rate of 60.00% (18/30) and 96.00% (24/25), respectively. The main color Doppler flow imaging (CDFI) feature of adult MASD was multiple colorful left-to-right shunt signals through the atrial septal designated, i.e. colander sign of CDFI. Conclusion TTE has some difficulties and TEE has specific value in diagnosing adult MASD. TTE can be used before open-chest operation. TEE is necessary before transcatheter occlusion to make sure of the amount and location of atrial septal defect.

Objective To investigate the relationship between antipituitary antibodies (APAs) and hypopituiarism following traumatic brain injury (TBI),as well as the severity of brain injury.Methods The study included 73 patients who suffered TBI 9 to 12 months ago and were diagnosed with hypopituitarism during the follow-up.Based on their Glasgow Coma Scale (GCS) on admission they were categorized into three groups:A (3-8),B (9-12) and C (13-15).Levels of plasma pituitary hormones (GH,TT,FT3 and FSH/LH) and APAs were measured in all patients with chemiluminescence assays and ELISA,respectively.Results Patients in group A had higher levels of APAs and lower levels of hormones compared with those in group B(P＜0.001) or group C(P＜0.001),while no significant difference was found between group B and group C for levels of either APAs (P＞0.05) or hormones (P＞0.05).Levels of APAs were negatively correlated with both GH (r=-0.64071,P＜0.001) and GCS (r=-0.50132,P＜0.001).Conclusion The present investigation provides preliminary evidence that APAs may be associated with the development of TBI-induced hypopituiarism.It suggests that the severity of hypopituiarism following TBI could be predicted by measuring the level of APAs.

Objective: To evaluate the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying the rmpA2-carrying hypervirulentKlebsiella pneumoniae. Methods: A total of 57nonduplicateKlebsiella pneumoniae isolates were collected from the Second Affiliated Hospital of Zhejiang University and Henan Provincial People′s Hospital. Virulence gene rmpA2 and capsule K serotype-specific genes were detected by PCR, multilocus sequence typing (MLST) was performed for molecular typing, and string test was conducted to identify the hypermucoviscous phenotype. The ClinProTools software was used for peak analysis. Four standard algorithms, including support vector machine (SVM), genetic algorithm (GA), supervised neural network (SNN), and quick classifier (QC), were tested for their power to differentiate between rmpA2-positive and rmpA2-negative strains. Results: Among the 57 Klebsiella pneumoniae isolates, 28 isolates belonged to sequence type (ST) 11 and carried the virulence gene rmpA2, of which 5 isolates were positive for string test; while the other 29 isolates were not detected rmpA2, and MLST divided them into five STs including ST11 (n=23), ST15 (n=3), ST1 (n=1), ST76 (n=1), and ST473 (n=1). The results of four standard algorithms were similar, while the sensitivity and specificity of SVM was the highest at 93.23% and 100% respectively. Analysis results of ClinProTools software suggested that the specific peaks for differentiating the rmpA2-positive and rmpA2-negative strains were 7 168.9 and 7 280.76, however, the peak intensity of themwere variant. Conclusion The sensitivity and specificity ofMALDI-TOF MS for rapid identification of rmpA2-carrying hypervirulentKlebsiella pneumoniae werehigher than 90% and there was a difference on the peak intensity of two specific peaks, which needs further study.(Chin J Lab Med, 2018, 41: 571-675)

Objective To evaluate the feasibility of using neutrophil bactericidal activity assay for analyzing the anti-bactericidal ability of hypervirulent Klebsiella pneumoniae ( hvKP) strains that harbored the virulence genes of rmpA and rmpA2 and were positive for string test .Methods A total of 150 non-duplicate blood-borne Klebsiella pneumoniae strains were collected from Zhejiang Province from January 2016 to July 2017.PCR was performed to detect carbapenem resistance genes (blaKPC, blaNDM, blaIMP-1, blaIMP-2), cap-sule genotypes (K1, K2, K5, K20, K54 and K57) and virulence genes (rmpA, rmpA2, iucA and iroN). Klebsiella pneumoniae strains that were positive for string test and harbored rmpA and rmpA2 genes were iden-tified as hvKP strains, while classic Klebsiella pneumoniae (cKP) strains were negative for string test, rmpA or rmpA2 gene.Neutrophil bactericidal activity assay was performed to analyze the virulence of Klebsiella pneumoniae strains and the survival rate was determined by using the following equation: the number of colony-forming units ( CFUs) in experimental group divided by the number of CFUs in control group .Re-sults Of the 150 Klebsiella pneumoniae strains, 43.3％ (65/150) harbored the rmpA2 gene and among them, strains positive for genes of rmpA, iroN and blaKPC and K2 respectively accounted for 73.8％, 80.0％, 75.4％and 40.0％.Twenty-four (36.9％) rmpA2 gene-positive strains showed positive result of string test.The survival rates of hvKP and cKP groups were respectively 0.866±0.056 and 0.368±0.058 and the difference between them was statistically significant (P<0.001).Conclusion Most of the hvKP strains that carry rmpA and rmpA2 genes and are positive for string test in Zhejiang Province survive the neu-trophil treatment , which indicates that the neutrophil bactericidal activity assay is an effective and simple method for identifying the virulence of Klebsiella pneumoniae.

Objective To investigate the effect of continuous nursing based on transtheoretical model (TTM) on elderly patients with knee osteoarthitis (KOA). Methods A total of 120 elderly patients with KOA from January 2015 to December 2016 were selected and divided into observation group and control group by the random digits table method , with 60 cases in each. The observation group was given continuous nursing based on TTM, while the control group was given continuous nursing. The functions and symptoms of knee joint of the 2 groups were evaluated by Lysholm Knee Scale Scores (LKSS) and Lequesne's Indexes (LI) after 6 months of implementation and compared. The compliance of rehabilitation training, the distribution of different behavior stages and the quality of life were compared after 6 months of implementation. Results The score of LKSS of the observation group was (78.04±8.55) points, and that of the control group was (69.22 ± 7.90) points, and there was significant difference between the 2 groups (t=6.780, P<0.05). The LI of the observation group was (5.37±1.05) points, and the control group was (6.09 ± 1.60) points, and there was significant difference between the 2 groups (t =3.356, P<0.05). There were significant differences in the distributions of the compliance of rehabilitation training and different behavior stages between the 2 groups (Z=5.814, 6.910, P<0.05). The rate of completely compliance, the percentage of action and maintenance stages of the observation group was 85.00%(51/60), 53.33% (32/60), and the control group was 55.00% (33/60), 28.33% (17/60) , and there was a significant difference between the 2 groups (χ2=12.857, 7.761, P<0.05). The physical condition, symptom and other 5 life quality scores and the total score of the observation group was (26.37±3.08), (11.80±2.15), (18.66± 2.27), (14.98±2.52), (6.93±0.94), (78.69±10.36) points, and the control group was (24.22±3.32), (9.44± 1.29), (17.24±3.07), (14.06±2.07), (6.56±1.05), (71.62±12.15) points, and there was significant difference between the 2 groups (t =2.034-7.291, all P<0.05). Conclusions The application of continuous nursing based on TTM on elderly patients with KOA can significantly change the stages of healthy behavior, improve the patients' compliance of rehabilitation training and quality of life, improve the function and symptom of knee joint.