AIM:To study the effect of integrin β1 on multidrug resistance in gastric cancer and its possible mechanisms .METHODS:The expression of integrin β1 at mRNA and protein levels in the SGC-7901 cells and SGC-7901/DDP cells was determined by qPCR and Western blot .The expression of integrin β1 in the SGC-7901/DDP cells was silenced by antisense oligodeoxynucleotide .The cell viability was detected by the CCK-8 assay, the cell apoptosis were ana-lyzed by flow cytometry, and the protein levels of integrin β1, Bcl-2/Bax, cleaved caspase-3/caspase-3, cytochrome C ( Cyt-C) and p-AKT/AKT were determined by Western blot .RESULTS:The expression of integrin β1 at both mRNA and protein levels was significantly upregulated in SGC-7901/DDP cells.The expression of integrin β1 was increased in SGC-7901 cells treated with chemotherapeutic agents such as cisplatin , paclitaxel and 5-fluorouracil .Knockdown of integrin β1 induced apoptosis of SGC-7901/DDP cells with an increased sensitivity to the chemotherapeutic agents .Meanwhile , knock-down of integrin β1 downregulated the protein levels of Bcl-2/Bax, p-AKTSer473 and p-AKTThr308 , while promoted the release of Cyt-C and upregulated the protein level of cleaved caspase-3.CONCLUSION:Knockdown of integrin β1 increases the sensitivity of SGC-7901/DDP cells to the chemotherapeutic agents , and promotes the cell apoptosis via mitochondrial apop-tosis pathway .The mechanism may be related to the attenuation of AKT pathway by inhibiting phosphorylations of AKT at Ser473 and Thr308.

Abstract: Objective　To analyze the etiological characteristics and drug resistance of patients with bloodstream infection (BSI) in the bacterial resistance monitoring network in Hainan Province from 2018 to 2020, so as to provide laboratory data for clinical diagnosis and treatment. Methods　The clinical data of the subjects were collected, and the etiological characteristics of BSI patients and drug resistance of commonly used drugs in clinical treatment were analyzed retrospectively. SPSS 26.0 software was used for statistical analysis. Results　A total of 877 strains were isolated, including Gram-negative bacteria (584 strains, 66.6%), Gram-positive bacteria (239 strains, 27.2%) and fungi (54 strains, 6.2%); male patients (591 cases, 67.4%), female patients (286 cases, 32.6%); inpatients (780 cases, 88.9%), outpatient and emergency patients (97 cases, 11.1%); the main primary diseases of BSI patients were hypertension, cerebral infarction and type 2 diabetes, and the main primary infections were pulmonary infection and urinary system infection. Intensive care unit (25.2%, 221 cases), emergency department (10.9%, 96 cases), oncology department (9.1%, 80 cases), nephrology department (6.8%, 60 cases) and hepatobiliary and pancreatic surgery department (4.3%, 38 cases) had the highest proportion of pathogenic bacteria. Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, coagulase-negative Staphylococcus, Viridans group streptococci and Candida albicans were the most frequently isolated pathogens. The detection rates of carbapenem-resistant Klebsiella pneumoniae, carbapenem-resistant Pseudomonas aeruginosa and carbapenem-resistant Acinetobacter baumannii were 3.4%, 15.2% and 36.4% respectively. The carbapenem-resistant Escherichia coli was not checked out. The detection rates of methicillin resistant Staphylococcus aureus and methicillin resistant coagulase negative Staphylococcus were 18.5% and 79.1% respectively. Conclusions　Gram-negative bacteria are the most common pathogens of BSI, and inpatients are the main source of BSI. Age, underlying diseases and primary infection are the risk factors of BSI. Clinical laboratories should strengthen the etiological monitoring of high-risk patients with BSI, and the resistance analysis of common antibiotics can provide a basis for the rational use of antibiotics in clinical practice.

Objective To detect the expression level of aquaporin 8 (AQP8) in patients with functional constipation(FC) or constipated irritable bowel syndrome (IBS-C),and the correlation between the expression of AQP8 and clinical features.Methods From March to December 2014,a total of 16 patients with IBS-C and 19 patients with FC met Rome Ⅲ criteria were collected,and nine healthy individuals were assigned to control group at the same period.The ascending and decending colonic tissues mucosa of FC,IBS-C and control group were taken under endoscope.The expression of AQP8 at mRNA and protein level was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).The differences in AQP8 mRNA expression and AQP8 relative area were analyzed by Kruskal-Wallis test among groups,and Pearson correlation coefficient was performed for correlation analysis between the expression and clinical features.Results The relative expressions of AQP8 mRNA of ascending colon and descending colon of FC patients (1.38,0.61 to 4.09;2.65,0.82 to 7.52) and IBS-C patients (2.23,0.82 to 4.67;1.35,0.51 to 2.03) were higher than those of control group (0.56,0.19 to 0.97;0.38,0.21 to 1.19),and the differences were statistically significant (ZFc =-2.435,-3.149,ZIBS-C =-2.690,-2.152;all P＜0.05).AQP8 mRNA expression of descending colon in patients with FC was higher than that of patients with IBS-C,and the difference was statistically significant (Z =-2.003,P =0.045).The expression of AQP8 in patients with FC and IBS-C was positively correlated with disease course (ascending colon r=0.57 and 0.53;descending colon r=0.49 and 0.54,all P＜0.05),and was negatively correlated with frequency of defecation (ascending colon r=-0.82 and-0.61;descending colon r=-0.49 and-0.53,all P＜0.05).There was no correlation between the expression of AQP8 and age,gender,onset age,presence of abdominal symptoms of the patients (all P＞ 0.05).Most of AQP8 of FC group was expressed in cytoplasm of colonic mucosa epithelial cells,while that of IBS-C group and control group was mostly expressed at apical membrane and basal membrane of epithelial cells.The results of semi-quantification demonstrated that AQP8 relative area of descending colon of FC and IBS-C group increased compared with that of control group (3.42％ (1.24％ to 5.61％),2.45％(1.72％ to 4.27％) vs 1.18％ (0.35％ to 2.81％);Z=-2.534,-2.151,both P＜0.05).Meanwhile,AQP8 relative area of ascending colon of FC group increased compared with that of control group (2.46％(1.48％ to 4.18％) vs 1.14％(1.29％ to 2.15％) Z=-2.041,P＜0.05).Conclusion There are differences in AQP8 expression quantity and location in cells of descending colon between patients with FC and patients with IBS-C,which is a way for differentiation these two diseases.

Adult ; Dermatitis, Occupational ; etiology ; Drug Eruptions ; etiology ; Humans ; Male ; Phthalimides ; adverse effects

Objective To summarize clinical experience of reconstructive operation with transpositional colon behind the sternum after corrosive esophageal burns and to explore the treatment for its complications.Methods Clinical data of 65 cases with esophageal scarred stricture after corrosive burns receiving reconstructive operation with transpositional colon behind the sternum were reviewed,56 of them by end-to-end anastomosis between transpositional anterograde peristaltic colon and esophagus,seven by end-to- end anastomosis between transpositional anterograde peristaltic colon and pharyngeal fundus,and two by end- to-end anastomosis between transpositional reversed peristaltic colon and esophagus,to summarize treatment experiences in pre-operation,operation and post-operation.Results Fifty-one of this group of patients recovered and discharged form the hospital smoothly,12 with cervical anastomotic leakage after operation including two cured by re-operation and ten cured by conservative treatment,and two with necrosis of transpositional colon including one died during operation and the other cured.Conclusions Corrosive burns of esophagus can be cured by leaving scarred stricture esophagus open without resection,and the effectiveness of reconstructive operation with transpositional colon behind the sternum is satisfactory with good pre-operative preparation,correct surgical operation,and correct post-operative treatment.

OBJECTIVETo clone Helicobacter pylori flagellin B gene (flaB) to construct prokaryotic expression system of the gene and to identify immunity of the fusion protein.

METHODSThe flaB gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The expression vector pET32a with inserted flaB gene was constructed. FlaB fusion protein was expressed in E.coli strain BL21DE3 inducted by IPTG at different dosages. Western blot was applied to determine immunoreactivity and immunogenicity of the fusion protein and antibody against whole cell of H.pylori and rabbit antiserum immunized with the fusion protein, respectively.

RESULTSIn comparison with the reported corresponding sequences, the homology of nucleotide sequence of the cloned flaB gene was from 96.31% approximate, equals 97.73%, while the homology of its putative amino acid sequence was as high as 99.41% approximate, equals 100%. The expression output of FlaB fusion protein in pET32a-flaB-BL21DE3 system was approximately 40% of the total bacterial proteins. FlaB fusion protein was able to combine with antibody against whole cell of H.pylori and induce rabbit to produce specific antibody with high titer after the animal was immunized with the protein.

CONCLUSIONA prokaryotic expression system of H. pylori flaB gene with high efficiency has been established successfully. The expressed FlaB fusion protein with satisfactory immunogenicity and immunoreactivity can be used as antigen in H.pylori vaccine and detect kit.

Antibodies, Bacterial ; blood ; Bacterial Vaccines ; immunology ; Base Sequence ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Flagellin ; genetics ; immunology ; Helicobacter pylori ; immunology ; Humans ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo study the effects of monocyte-macrophages (THP-1) in malignant transformation of human bronchial epithelial cells (BEAS-2B) cells induced by coal tar pitch (CTP) and the expression of TNF-α in the process of the cell malignant transformation.

METHODSBEAS-2B cells and THP-1 Cells were divided into four groups: coal tar pitch (CTP) group, benzo(a)pyrene [B(a)P] group, dimethyl sulfoxide (DMSO) group, BEAS-2B and THP-1 co-culture (co-culture group) group. Carcinogenesis model was established. The soft agar colony formation, chromosome aberrations and cell cycle tests were used to detect the cellular malignant transformation. The ELISA assay was utilized to measure the levels of TNF-α in the supernatant of CTP group and co-culture group.

RESULTSThe chromosome number abnormalities could be observed in early stage of the experiment (the 10th generation cells), which showed the increased ratio of aneuploid to polyploid, and the decreased number of diploid. The colony formation rate of co-culture group (the 20th generation cells) was 17.63‰ ± 0.97‰, which was significantly higher than that (13.94‰ ± 0.84‰) of CTP group and that (12.96‰ ± 1.62‰) of B(a)P group (P < 0.05). The proportion of S phase cells in the co-culture group was 44.49% ± 0.68%, which was significantly higher than that (38.19% ± 1.26%) of CTP group and that (36.41% ± 1.19%) of B(a)P group (P < 0.05). The TNF-α level in the co-culture group was significantly higher than that in CTP group (P = 0.001).

CONCLUSIONMonocyte-Macrophages can accelerate the malignant transformation of BEAS-2B cells induced by CTP and increase the expression level of TNF-α.

Bronchi ; cytology ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; Coal Tar ; toxicity ; Coculture Techniques ; Epithelial Cells ; cytology ; drug effects ; pathology ; Humans ; Macrophages ; cytology ; Monocytes ; cytology ; Tumor Necrosis Factor-alpha ; metabolism

Objective To evaluate the reliability and validity of the application of condensed Chinese version of the MOS 36-item short form health survey (SF-36) in assessment of quality of life among adult patients with Kashin-Beck disease,and to provide a scientific basis in rehabilitation of the patients.Methods Four hundred and twenty seven eases of adult patients with Kashin-Back disease and 419 healthy individuals randomly selected in Kashin-Beck disease endemic areas in 8 counties of Gansu province were surveyed with the SF-36.The reliability of the SF-36 was assessed by split-half reliability and Cronbach's α coefficient and the validity through principal component factor analysis and correlation analysis,etc.The dimension scores of different people were obtained by analysis of variance and univariate t-test.Results The split-half reliability of all the 8 dimensions was greater than 0.6 and the Cronbach's α coefficient was greater than 0.8; the pearson correlate coefficients of all the items to their dimensions were greater than 0.391.SF-36 contained 8 domains and 2 summary scales in the factor analysis.The score differences of quality of life in different ages of the patients,different stages of the disease were statistically significant (all P ＜ 0.05).Conclusion The SF-36 is practical in studying the quality of life among adult patients with Kashin-Beck Disease.

Primary central nervous system germ cell tumors (CNS-GCTs) in children and adolescents have unique clinical features and methods of treatment compared with those in adults. There is little information about Chinese children and adolescents with CNS-GCTs. Therefore, in this study we retrospectively analyzed the clinical features and treatment outcome of Chinese children and adolescents with primary CNS-GCTs. Between January 2002 and December 2012, 57 untreated patients from a single institution were enrolled. They were diagnosed with CNS-GCTs after pathologic or clinical assessment. Of the 57 patients, 41 were males and 16 were females, with a median age of 12.8 years (range, 2.7 to 18.0 years) at diagnosis; 43 (75.4%) had non-germinomatous germ cell tumors (NGGCTs) and 14 (24.6%) had germinomas; 44 (77.2%) had localized disease and 13 (22.8%) had extensive lesions. Fifty-three patients completed the prescribed treatment, of which 18 underwent monotherapy of surgery, radiotherapy, or chemotherapy, and 35 underwent multimodality therapies that included radiotherapy combined with chemotherapy or surgery combined with chemotherapy and/or radiotherapy. PEB (cisplatin, etoposide, and bleomycin) protocol was the major chemotherapy regimen. The median follow-up time was 32.3 months (range, 1.2 to 139 months). Fourteen patients died of relapse or disease progression. The 3-year event-free survival (EFS) and overall survival rates for all patients were 72.2% and 73.8%, respectively. The 3-year EFS was 92.9% for germinomas and 64.8% for NGGCTs (P = 0.064). The 3-year EFS rates for patients with NGGCTs who underwent monotherapy and multimodality therapies were 50.6% and 73.5%, respectively (P = 0.042). Our results indicate that multimodality therapies including chemotherapy plus radiotherapy were better treatment option for children and adolescents with CNS-GCTs.
Adolescent ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; Central Nervous System Neoplasms ; therapy ; Child ; Child, Preschool ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; statistics & numerical data ; Disease-Free Survival ; Etoposide ; administration & dosage ; Female ; Humans ; Male ; Neoplasm Recurrence, Local ; Neoplasms, Germ Cell and Embryonal ; therapy ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo obtain monoclonal antibodies (mAb) against LI-cadherin and investigate their effects on the proliferation of human hepatocellular carcinoma cells.

METHODSBalb/c mice were immunized with recombinant LI-cadherin, and hybridoma cell lines secreting monoclonal antibodies against LI-cadherin were established with routine cell fusion and subcloning approach. The specificity of these mAbs was determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The effect of the mAbs obtained on the growth of HepG2 cells was assessed using inverted microscope and MTT assay.

RESULTSTwo hybridoma cell lines (F001 and F002) stably secreting specific mAbs were obtained. Western blot analysis showed that the two antibodies specifically recognized LI-cadherin antigen derived from human eucaryotic cells or tissue. Treatment of the HepG2 cells with the mAbs resulted in reduced viable cell number and changes in the cell morphologies, and the two mAbs inhibited the proliferation of HepG2 cells in a concentration-dependent manner (P<0.05).

CONCLUSIONThe two specific mAbs obtained can inhibit the proliferation of HepG2 cells in vitro, which facilitates further study of the relationship between LI-cadherin and tumors.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; pharmacology ; Cadherins ; immunology ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Hybridomas ; secretion ; Liver Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C