Objective To isolate and purify VacA protein secreted by Helicobacter pylori or recombinant VacA , and to investigate the effect of VacA-induced cell vacuolar change and apoptosis .Methods VacA proteins were separated and pu-rified from the culture supernatant of H.pylori ( ATCC26695 ) or from the split products of genetically engineered bacteria (pQE30-VacA-E.coli M15) expressing recombinant VacA.The VacA protein obtained was acidified and then incubated with AGS cells for 24 h at different final concentrations of 5 and 10 ng/ml before the vacuolar change and apoptosis of AGS cells were detected via microscopy and flow cytometry assay , respectively .Results H.pylori-secreted VacA and recombi-nant VacA were successfully separated and purified .The H.pylori-secreted VacA significantly induced the vacuolar change and apoptosis of AGS cells (P<0.01) while the recombinant VacA did not.Conclusion H.pylori-secreted VacA protein can effectively induce cell vacuolar change and apoptosis, but recombinant VacA can not, suggesting that the purified VacA protein secreted by H.pylori can be used to explore VacA-induced pathogenesis.

Objective To investigate the correlation between Helicobacter pylori (HP) infection-associated gastritis and the ap-optotic genes in gastric mucosa .Methods Forty-five patients with chronic gastritis were registrated in our study from November 2013 to December 2014 .HP infection status in the patients was detected by using urease test and 13C-urea breath test .The degree of gastritis in the gastric mucosa with HP infection was confirmed via histopathology .qRT-PCR was used to measure the mRNA ex-pressions of Bax ,Bak and Bcl-2 in the gastric mucosa with HP infection and matched normal gastric mucosa .Person analysis was used to assess the correlation between the HP infection-associated gastritis and the mRNA expressions of Bax ,Bak and Bcl-2 in the gastric mucosa .Results Forty-five patients with HP infection in antrum and 45 patients (100% ) with chronic antrum gastritis were identified ,including 28 patients (62 .2% ) with light gastritis ,16 patients (35 .6% ) with moderate gastritis ,1 patient (2 .0% ) with severe gastritis .9 patients (20 .0% )with metaplasia ,5 patients(11 .1% ) with low grade intraepithelial neoplasms .The urease tests were negative in the gastric body of 45 patients ,6 patients (13 .3% )were mild chronic gastritis in the body ;Patient with meta-plasia and intrapithelial gastritis was not found .The Bax expression in the HP-infected gastric mucosa was markedly increased when compared with the normal gastric mucosa (P< 0 .01) ,and positively correlated with the degree of gastritis (P< 0 .01) , whereas the expressions of Bak and Bcl-2 have no significantly deferences bttween two groups(P>0 .05) .Conclusion HP infec-tion-associated gastritis positively correlated with the expressions of apoptotic genes in gastric mucosa ,suggesting that HP infection might result in increasing the Bax expression and further enhancing the cell apoptosis .

0.05).We did not find any difference of the expression of fibronectin,laminin and type IV collagen in them.Expression of ICAM and VCAM were(56.4?14.8)% and(55.6?12.2)%,respectively,obviously higher than those in control group(P

Objective To detect the mutation of ATP7A gene in 2 families with Menkes disease.Met-hods Genomic DNA of 6 members from 2 families were extracted with salt fractionation.The encoding exons and 2 sides flanking intron of ATP7A gene were amplified from genomic DNA of the probands and their parents by PCR and directly sequence.Light microscope was used to test the hair of probands and normal healthy children.Results Proband 1 had a deletion mutation of c.3 045del T in exon 14 of ATP7A gene and resulted in a stop codon just several nucleotides behind the deletion site.His mother was a heterozygote of the mutation and had normal phenotype.Proband 2 had a nonsense mutation of c.2 956 in exon 14 of ATP7A gene and resulted in a stop of amino acid synthesis.His mother was not a heterozygote of the mutation.Genetype and phenotype in fathers of the 2 probands were normal.Hair of the probands in light microscope were tenuity,midheaven,the color of hair also turned to light.Conclusions The c.3 045del T mutation of ATP7A gene cause the phenotype of Menkes disease in proband 1.His mother is a heterozygote of the disease without symptoms and he is of familial inheritance.The c.2 956 nonsense mutation of ATP7A gene cause the phenotype of Menkes disease in proband 2.His mother is not a heterozygote of the disease and he is not of familial inheritance.

To investigate the expression of NF-kappaB in acute leukemia and its relationship with P21, and matrix metalloproteinases (MMP), the expression of NF-kappaB, P21, MMP-2 and MMP-9 in bone marrow cells from patients with acute leukemia (AL) was detected using immunocytochemical technique. The results showed that the expression ratios of NF-kappaB, P21, MMP-2 and MMP-9 in untreated AL group were significantly higher than those in remission and normal control groups (P < 0.05), and no obvious difference was seen between remission and normal control groups. The expression of NF-kappaB was correlated with that of P21, MMP-2 and MMP-9 (r = 0.767, 0.729 and 0.803, respectively, P < 0.05). This study indicated that P21 protein, encoded by oncogene Ras, and NF-kappaB were super-expressed in leukemia cells. In conclusion, after activation by Ras, NF-kappaB combined with the kappaB sequences of MMP-2 and MMP-9 genes, then upregulated their expression. MMP might enhance the degradative function of leukemic cell, thus to make cells easier to cross through the bone marrow barrier and release into blood.
Acute Disease ; Adult ; Bone Marrow Cells ; metabolism ; Female ; Humans ; Immunohistochemistry ; Leukemia ; drug therapy ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; Matrix Metalloproteinase 9 ; biosynthesis ; NF-kappa B ; biosynthesis ; Proto-Oncogene Proteins p21(ras) ; biosynthesis ; Remission Induction

Adenosine Triphosphatases ; genetics ; Cation Transport Proteins ; genetics ; Copper-transporting ATPases ; Gene Deletion ; Humans ; Infant ; Male ; Menkes Kinky Hair Syndrome ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the clinical value of pleural effusion lung ProGRP, neuron specific enolase (NSE), cytokeratin fragment 19 (CYFRA21-1), carcino-embryonic antigen (CEA), carbohydrate antigen 153 (CA153), carbohydrate antigen 19 - 9 (CA19-9) in differential diagnosis and histological typing of malignant pleural effusion caused by lung cancer.

METHODSAll the 171 patients with malignant hydrothorax caused by lung cancer were from coal-mine area of Kailuan. They were divided into the small cell lung cancer (SCLC) group (n = 39), the adenocarcinoma group (n = 99) and the squamous cell carcinoma group (n = 37). The patients with benign pleural effusion served as the controls (n = 30). The diagnostic value of pleural effusion ProGRP, NSE, CYFRA21-1, CEA, CA153 and CA19-9 was compared for each group.

RESULTSYouden index and the accurate rate of pleural effusion ProGRP + NSE (sequence test) were the highest in the diagnosis of malignant hydrothorax caused by SCLC. CEA + CA153 + CA19-9 (sequence test) was the highest in the diagnosis of malignant hydrothorax caused by adenocarcinoma. CYFRA21-1 + CEA + CA153 (on parallel test) were the highest in the diagnosis of malignant hydrothorax caused by squamous cell carcinoma. The Yonden index and the accurate rate were the highest by the single detection of CYFRA21 (0.5514 and 0.6878), and by the combined detection of ProGRP + CYFRA21-1 + CEA (on parallel test) (0.7029 and 0.8878).

CONCLUSIONThe first pleural effusion tumor markers of malignant hydrothorax caused by the SCLC, adenocarcinoma of lung, and lung squamous cell carcinoma are ProGRP, CEA and CYFRA21-1, respectively. The best combinations of pleural effusion tumor marker in diagnosis of malignant hydrothorax caused by the SCLC, adenocarcinoma of lung, lung squamous cell carcinoma and lung cancer are the combined detection of ProGRP + NSE (sequence test), combined detection of CEA + CA153 + CA19-9 (sequence test), the combined detection of CYFRA21-1 + CEA + CA153 (on parallel test) and ProGRP + CYFRA21-1 + CEA (on parallel test), respectively.

Adult ; Aged ; Aged, 80 and over ; Antigens, Neoplasm ; analysis ; Biomarkers, Tumor ; analysis ; CA-19-9 Antigen ; analysis ; Diagnosis, Differential ; Female ; Humans ; Keratin-19 ; analysis ; Lung Neoplasms ; complications ; diagnosis ; Male ; Middle Aged ; Peptide Fragments ; analysis ; Pleural Effusion, Malignant ; diagnosis ; etiology ; Recombinant Proteins ; analysis

Background: and Purpose Mutations in the FIG4 gene have been linked to amyotrophic lateral sclerosis (ALS) type 11 in Caucasian populations. The purpose of this study was to identify FIG4 variants in a cohort of 15 familial ALS (FALS) indexes and 275 sporadic ALS (SALS) patients of Han Chinese origin. Methods: All 23 exons of FIG4 were sequenced using targeted next-generation sequencing.An extensive literature review was performed to detect genotype-phenotype associations of FIG4 mutations. Results: No FIG4 variants were identified in the FALS patients. One novel heterozygous missense variant (c.352G>T [p.D118Y]) and one novel heterozygous nonsense variant (c.2158G>T [p.E720X]) in FIG4 were identified in two SALS patients. The p.E720X variant is interpreted as likely pathogenic while the p.D118Y variant is a variant of uncertain significance. The patient carrying the p.E720X mutation developed lower-limb-onset slowly progressive ALS, and survived for 11.5 years. The patient harboring the FIG4 p.D118Y variant also presented with progressive ALS, with the score on the ALS Functional Rating Scale–Revised (ALSFRS-R) decreasing by 0.4 per month. The rate of decrease in the ALSFRS-R scores from symptom onset to diagnosis seemed to be lower in the patients carrying FIG4 variants than the no-FIG4-mutation ALS patients in this study. Conclusions Our findings suggest that ALS patients carrying FIG4 mutations are not common in the Chinese population and are more likely to exhibit slow progression.

Objective To research the sequencing weaning in the patients with chronic obstructive pulmonary disease (COPD). Methods The COPD patients were divided into two groups in this prospective case-control study: protocol-directed group vs none- protocol-directed group. The patients in the former group received programmed ventilator weaning strategy, detaily patients were selected through the shallow breathing with fast exponential screening, then were conducted by spontaneous breathing trial SBT , finally were pulled out the endotracheal intubation. The patients in the latter group were determined the timing of ending the ventilator according to doctors subjective judgment and clinical experience. Results The ventilation time of protocoldirected and on-protocol-directed group was ( 8.01 ± 7. 22 ) and ( 10.74 ± 5.58 ) days ( P = 0. 034 ) respectively. ICU stay time was shorter in protocol-directed group than in non-protocol-directed group ( 8.86 ±6.92 vs 12 ±6.10 days, P =0. 026). The success rate of weaning in protocol-directed group was 76.31% (29/38),which was higher than that in non-protocol ～ directed group( 12/25,48% , P = 0. 009 ). Conclusions Protocoldirected weaning and SBT may reduce the ventilation time and ICU stay time and enhance success rate of weaning.

Objective To investigate present situation of ICU nurses' job burnout,self-efficiency and capacity for empathy and explore the relationship among them.Methods The Chinese Maslach Burnout Inventory,General Self-efficacy Scale and the Jefferson Scale of Empathy-Health Professionals were used to conduct a survey among 220 ICU nurses in Provincial Third-level Comprehensive Hospitals in Changsha.Descriptive statistics and Pearson correlation analysis were applied.Results Totals of 220 questionnaires were handed out and 215 were recovered,with the efficiency rate of 97.72％.The score of 215 ICU nurses' emotional exhaustion and depersonalization was respectively (23.28 ± 9.72) and (6.40 ± 5.72),which were moderate burnout.The score of personal accomplishment was (32.27 ± 9.36),a high degree of burnout.The average score of self-efficiency was (2.54 ± 0.51) and the average score of capacity for empathy was (110.33 ±12.36),in which the average score of perspective taking,compassionate care and ability to stand in patients' shoes was (56.64 ±8.02),(40.32 ±6.34) and (13.36 ±2.30) respectively.Emotional exhaustion had significantly negative correlation with self-efficiency,capacity for empathy and ability to stand in patients' shoes (P ＜ 0.05).Depersonalization had significantly negative correlation with self-efficiency,capacity for empathy and its three dimensions (P ＜ 0.05).Personal accomplishment had significantly positive correlation with self-efficiency,capacity for empathy and its three dimensions (P ＜ 0.05).Conclusions The problem of job burnout is serious in ICU nurses.Special attention should be paid to increase their self-efficiency and capacity for empathy so as to reduce their job burnout.