OBJECTIVETo establish a rapid specific method to identify the single-nucleotide polymorphisms (SNPs) of HBV polymerase gene region which are the methionine residue of the conserved YMDD motif.

METHODSTwo specific primers were designed to amplify interested gene region involved in SNPs which were also used as HBV DNA identification. Specific primers of SNaPshot were designed to detect 741A-G (YVDD), 743G-T (YIDD). The different fluorescent dye labeled ddNTP was used to further extend the strand of PCR product and was detected by ABI PRISM 310 Genetic Analyzer. Sera from 13 patients with chronic hepatitis B after lamivudine treatment were analyzed.

RESULTSAside from mutation of YMDD, there were mutations of 514C-A, 523C-A, 562T-A, 667C-A. The 13 samples were simultaneously tested with SNaPshot and DNA sequencing, the same results were obtained. The method of SNaPshot showed high specificity.

CONCLUSIONMutation of YMDD results in the changes of ATG codon, and there are new ATG codon in the upper strand of YMDD. SNaPshot technique is rapid, specific and accurate for the SNPs monitoring of HBV DNA mutation during lamivudine therapy. Two samples were determined by SnaPshot technique, identifying the co-existence of the mixed wild type and mutant type HBV infection.

Antiviral Agents ; therapeutic use ; Base Sequence ; DNA, Viral ; blood ; genetics ; Drug Resistance, Viral ; Gene Products, pol ; genetics ; Hepatitis B ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVETo study the protective effect of limb ischemic preconditioning against liver ischemia/reperfusion injury in the rat.

METHODSRats were randomly divided into four groups (n = 8): (1) Sham group (S group), rats without ischemic preconditioning (IPC), (2) Ischemia/reperfusion (I/R) without IP (I/R group); (3) Rats with 5 min IPC (IPC group); (4) Rats with lower limbs IPC and repeated three times (remote ischemic preconditioning, RPC group); The rats were subjected to 60-min sustained liver ischemia followed by 180-min reperfusion except S group. All ischemia rats were only subjected to 70% liver ischemia. Finally, blood and liver samples were obtained to determine the activity of ALT and AST, liver wet/dry weight (W/D), PMN counts and pathology.

RESULTSAll IPC group and RPC group had obviously lower levels of ALT, AST, W/D, PMN counts than that of the I/R group (P < 0.01).

CONCLUSIONThe limb ischemic preconditioning has a protective effects against liver ischemia/reperfusion injury in the rat, possibly are due to suppression of liver inflammatory reaction, improvement of liver microcirculation.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Ischemic Preconditioning ; Liver ; blood supply ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control

OBJECTIVE: To investigate the clinical characteristics of essential thrombocytopenia (ET) patients with positive mutations including JAK2, CALR, MPL, or negative mutations. METHODS: A total of 66 newly diagnosed ET cases from January 2016 to December 2018 in Department of Hematology, Huaian No.1 People's Hospital affiliated to Nanjing Medical University were analyzed. Statistical analysis data included the patient's sex, age, symptoms, thrombosis and embolism events, spleen omegaly, platelet count (Plt), leukocyte (WBC) count, hemoglobin (Hb), fibrinogen (FIB), thrombus elastic diagram (TEG), serum potassium, blood glucose (GLU), lactate dehydrogenase (LDH), JAK2, CALR and MPL mutations, treatment options, and efficacy. RESULTS: All the patients were not MPL-positive, and divided in three groups: JAK2 mutation (46 cases, 69.7%), CALR mutation (9 cases, 13.6%) and gene negative mutation (11 cases, 16.7%) group. The average age of patients in the JAK2 mutation group was 63.2 years old, and significantly higher than that in the CALR mutation group (51.8 year) and gene negative group (50.2 year) (P＜0.05). Compared with the JAK2 mutation group and gene negative group, the CALR mutation group had lower WBC count (6.3×10/L vs 13.79×10/L) (P=0.003) (6.3×10/L vs 9.70×10/L) (P=0.009). Also the Hb level of patients in CALR mutation group was lower than the JAK2 mutation group (121.22 g/L vs 136.2 g/L) (P=0.036). However, there was higher tumor burden in the CALR mutation group, compared with the gene negative mutation group (300.11 U/L vs 227.4 U/L) (P=0. 033). There was no significant difference among the three groups, such as the Plt counts, serum potassium level, GLU level and FIB level (P＞0.05). In addition, thrombus and embolism appeared in 30.3% (20/66) cases. 18.2% (12/66) cases were complicated with hyperkalemia, which significantly correlated with Plt counts (r=0.518). TEG was performed in 34 patients, of which 41.2% (14/34) had abnormal TEG and 55.9% (19/34) were accompanied by Plt count ＞ 1 000 ×10/L, but there was no significant correlation between them (r=0.134). After routine clinical treatment, all the 66 cases achieved partial or complete hematological remission, but the disease usually repeated. Until now 4.5% (3/66) cases had been converted to myelofibrosis (MF) all with JAK2 mutation, but without advancing to acute myeloid leukemia. CONCLUSION ET patients with JAK2 mutation have higher incidence, moreover were in older age. However, the patients with CALR mutations display lower WBC count and Hb level, but higher tumor burden. In short, the multiple gene mutations of ET showed different clinical features closely relates with the prognosis, thus providing guidance for the clinical diagnosis and treatment.
Aged ; Calreticulin ; genetics ; Humans ; Janus Kinase 2 ; genetics ; Middle Aged ; Mutation ; Primary Myelofibrosis ; Thrombocythemia, Essential ; Thrombocytopenia

OBJECTIVETo investigate the inhibitory effect of HSP90 inhibitory 17-AAG on proliferation of multiple myeloma cells and its main mechanism.

METHODSThe multiple myeloma cells U266 were treated with 17-AAG of different concentrations (200, 400, 600 and 800 nmol/L) for 24, 48, and 72 hours respectively, then the proliferation rate, expression levels of β-catenin and C-MYC protein, as well as cell cycle of U266 cells were treated with 17-AAG and were detected by MTT method, Western blot and flow cytometry, respectively.

RESULTSThe 17-AAG showed inhibitory effect on the proliferation of U266 cells in dose- and time-depetent manners (r = -0.518, P < 0.05 and r = -0.473, P < 0.05), while the culture medium without 17-AAG displayed no inhibitory effect on proliferation of U266 cells (P > 0.05). The result of culturing U266 cells for 72 hours by 17-AAG of different concentrations showed that the more high of 17-AAG concentration, the more low level of β-catenin and C-MYC proteins (P < 0.05); At same time of culture, the more high of 17-AAG concentration, the more high of cell ratio in G1 phase (P < 0.05), at same concentration of 17-AAG, the more long time of culture, the more high of cell ratio in G1 phase (P < 0.05).

CONCLUSIONThe HSP90 inhibitory 17-AAG can inhibit the proliferation of multiple myeloma cells, the down-regulation of Wnt/β-catenin signaling pathway and inhibition of HSP90 expression may be the main mechnisms of 17-AAG effect.

Apoptosis ; Benzoquinones ; pharmacology ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; pharmacology ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

Objective To investigate the significance of MTSS1 and E-cadherin expression in upper urinary tract transitional epithelial carcinoma.Methods Paraffin specimens of 60 patients with upper urinary tract transitional epithelial carcinoma between January 2005 and January 2014 were analyzed.At the same time,5 cm normal tissue adjacent to the cancerous tissue specimens in 30 patients were taken for comparison.Immunohistochemical method was used to detect the tissue MTSS1 and E-cadherin expression,and the relationships between their expression with different pathological stage,differentiated degree and lymph node metastasis were analyzed.Results MTSS1 expression rate in normal tissue (100.0％,30/30) was significantly higher than that in cancerous tissue (45.0％,27/60) and there was significant difference (P ＜ 0.05).E-cadherin expression rate in normal tissue (96.7％,29/30) was significantly higher than that in cancerous tissue (41.7％,25/60) and there was significant difference (P ＜ 0.05).The expression of MTSS1 and E-cadherin in different pathological stage,degree and with or without lymph node metastasis had significant difference (P ＜ 0.05).In patients with well differentiated,low TNM stage and no lymph metastasis,MTSS1 and E-cadherin expression rate was higher (P ＜ 0.05).There was no significant correlation between the expression of MTSS 1 and E-cadherin in cancerous tissue (P ＞ 0.05).Conclusion In upper urinary tract transitional cell carcinoma detection of both MTSS1 and E-cadherin has important significance with regards to judging the malignant degree of the tumor,lymph node metastasis and prognosis in patients.

Objective:To investigate the effect and related mechanism of TRAP1 on the invasion and migration of human bladder cancer through TGF/Smad3 signal path.Methods: Selected from BIU-87 of high expression of TRAP1 in bladder cancer cell lines through Western blot techniques.TRAP1 knockdown lentivirus (LV3-TRAP1) was used to silence the expression of TRAP1.GFP fluorescene and PCR detector was used to detected the efficiency of gene silencing and the effectiveness of gene silencing;effect of TRAP1 on the invasion and migration ability of BIU-87 were detected by Transwell matrigel invasion assays and wound healing assays,CM-H2DCFDA fluorescent staining was used to deteced the cell ROS of BIU-87 with LV3-TRAP1.Detected the level of TGF/Smad3 signal protein by Western blot.Results: LV3-TRAP1 lentivirus could effectively inhibit the expression of TRAP1 compared with LV3-NC.LV3-TRAP1 lentivirus could effectively inhibit the cell RPS of BIU-87.Knockdown the expression of TRAP1 could inhibit the invasion and migration of BIU-87.Knockdown the expression of TRAP1 in BIU-87 could reduce the protein level of TGF/Smad3.Conclusion: Silencing TRAP1 could inhibit the invasion and migration of bladder cancer cell through TGF/Smad3 signal pathway.

Objective: To analyze the status of job burnout among the community and township public health workers, and to provide scientific basis for formulating comprehensive prevention and control measures. Methods: A census sampling method was used to investigate the job burnout by using the self-made general demographic data questionnaire and the MBI-GS in HuaiAn. Results: A total of 1074 valid questionnaires were collected, and the total physical examination rate of job burnout was 58.7%, 51.3% were mild burnout, 7.4% were highly burnout. Multivariate ordered logistic regression analysis showed that sex (OR=1.32), chronic disease (OR=1.92)、and daily working hours greater than 7 h (OR=1.40)、township health center (OR=1.31) were independent risk factors for occupational burnout. The age "≥51" (OR=0.45)、"41~" (OR=0.58) are the independent protective factors for the occurrence of job burnout. Conclusion Job burnout detection rate was high in the staff of essential public health service, sex, daily working hours, health status, age and work unit are the main factors influencing job burnout.

OBJECTIVE: Apneagraph can be used to discuss which the best operation scheme is for OSAHS. Effects of uvulopalatopharyngoplasty can be assessed by Apneagraph in obstructive sleep apnea hypopnea syndrome (OSAHS) patients. METHOD: Fifty-six patients with OSAHS received the modified UPPP operation were randomly selected in our hospital. The AG and PSG were applied for diagnosis and evaluation of operation effects. The sleepiness state was assessed by ESS (Epworth sleepiness scale) 6 months after the surgery, compared with the preoperative ESS scores using attest for statistical analysis. We used the SPSS19.0 software to carry our data analysis. RESULT: After 6 months, the evaluation of postoperative efficacy came out to be completely controlled in 42 cases (75%), significantly effective in 14 cases (25%), and uncured in 0 cases. Correlation between the transpalatal obstruction proportion and the AHI reduction percentage was significantly positive (r = 0.667). There were 38 patients with oropharynx obstruction percentage more than 73.35% presented completely controlled in 34 cases (89.47%), significantly effective in 4 cases (10.33%), and uncured in 0 cases. CONCLUSION AG has the dual functions of analyzing sleep-related respiratory disturbance events and determining upper airway obstruction sites. AG application in the postoperative evaluation of modified uppp has significantly objective guide significance. The modified UPPP for treatment of OSAHS can improve the operation effect. Patients with oropharynx obstruction percentage more than 73.5% don't need to receive the operation for treatment of retroglottal region.
Adult ; Cleft Palate ; surgery ; Female ; Humans ; Male ; Middle Aged ; Palate ; surgery ; Palate, Soft ; surgery ; Pharynx ; surgery ; Polysomnography ; Sleep Apnea, Obstructive ; physiopathology ; surgery ; Sleep Stages ; Treatment Outcome ; Young Adult

BACKGROUNDInterferon-induced transmembrane protein 1 (IFITM1) has been identified as a molecular marker of the colorectal tumors; however its influences on the biological behaviors of the colorectal cancer cells are currently unknown. We aimed to study the influences of IFITM1 on the proliferation, invasion, and metastasis of the colorectal cancer SW480 cell lines.

METHODSWe constructed IFITM1/pEGFP-C3 recombinant plasmids and transfected them into the colorectal cancer SW480 cell lines. IFITM1/pEGFP-C3 recombinant plasmids were identified by means of immunofluorescence, laser confocal scanning microscopy, and reverse transcription polymerase chain reaction. IFITM1/SW480 cells with stable over-expression of IFITM1 were confirmed by G418 screening. The influences of IFITM1 on the proliferation of the SW480 cell lines were investigated by MTT assay and tumor transplantation experiments in nude mice. Cell invasion experiments were performed to determine the invasion capacity of the IFITM1/SW480 cells. Matrix metalloproteinase 2 (MMP-2) and MMP-9 activities were detected by the gelatin zymographic analysis, and MMP-9 expression by the Western blotting analysis.

RESULTSIFITM1/pEGFP-C3 recombinant plasmids were successfully constructed in this study, and the IFITM1/SW480 cells with stable IFITM1 gene over-expression were confirmed by G418 screening. MTT results showed that the proliferation of the IFITM1/SW480 cells was significantly enhanced (P < 0.01). Tumors were harvested from four weeks old mice. Tumor volumes were (1347.00 ± 60.94) mm(3), (1032.40 ± 111.38) mm(3) and (1018.78 ± 28.83) mm(3); and tumor weights were (1522.34 ± 62.76) mg, (1137.78 ± 97.22) mg and (1155.76 ± 133.31) mg for mice inoculated with the IFITM1/SW480 cells, pEGFP-C3/SW480 cells and SW480 cells, respectively. Tumor volumes and weights from mice inoculated with the IFITM1/SW480 cells were significantly increased (P < 0.01). In addition, the numbers of the SW480 cells and IFITM1/SW480 cells that migrated through Matrigel were 448.64 ± 38.09 and 540.45 ± 44.61, respectively; so the invasive ability of the SW480 cells transfected with IFITM1 gene was significantly greater than that of the SW480 cells (P < 0.01). Gelatin zymographic analysis showed that MMP-9 and MMP-2 protein activities in the IFITM1/SW480 cells were significantly enhanced, and Western blotting analysis showed that MMP-9 expression in the IFITM1/SW480 cells was also increased.

CONCLUSIONIFITM1 can enhance the proliferation, invasion, and metastasis of the colorectal cancer SW480 cell lines.

Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Plasmids ; Vesicular Transport Proteins ; genetics ; metabolism

Acute lymphoblastic leukemia (ALL) is a malignant clonal disease, its treatment methods include chemotherapy, hematopoietic stem cell transplantation, immunotherapy and molecular targeted therapy. Clinically, ALL patients need to get complete remission through chemotherapy, and then choose the other treatment according to the patient's condition. But the drug resistance has been a biggest obstacle in treatment of ALL. There are many research reports about drug-resistant of ALL at present. In this review, the classic drug resistance mechanisms, such as membrane transporter, gene modifications and some newly finding mechanisms including such as bone marrow microenvironment and Micro RNA and so on are summarized.
Bone Marrow ; physiology ; Cellular Microenvironment ; Drug Resistance, Neoplasm ; Humans ; Membrane Transport Proteins ; physiology ; MicroRNAs ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy