Objective To investigate the p53,Bax,bcl-2 gene in NaAsO2-induced human embryonic lung fibroblasts(HELF)apoptosis.Methods HELF was divided into HELF cells transfected with p53 plasmid(p53 group),HELF cells transfected with PC plasmid(PC group)and normal cultured HELF cells(normal group).The mRNA expression of p53,Bax and bcl-2 gene was detected by real-time PCR,the protein expression of p53,Bax and bcl-2 was assessed by immunohistochemical SABC and the cell apoptosis of HELF was detected by flow cytometry(FCM),in a 6-well plate and cultured for 48 hours,which was exposed to different doses(0,3,9,15mmol/L)NaAsO2 for 24 hours.Results The p53 gene mRNA expression level of p53 group(0.51±0.29)was lower than that of the normal group and PC group [ (1.00 ± 0.20), (1.32 ± 0.26), all P < 0.05 ]. The p53 protein expression level of p53 group(4.10 ± 1.20) was lower than the PC group and normal group[ (8.00 ± 1.63), (7.90 ± 1.79), allP < 0.05]. In p53 group, PC group, normal group exposed to 0,3,9,15 mmol/L NaAsO2 doses, the apoptotic rate [(0.57 ± 0.28)%, (22.91 ± 4.86)%, (40.05 ± 3.93)%, (44.87 ± 3.58)%; (0.65 ± 0.24)%, (14.09 ± 3.49)%,(20.31 ± 3.66)%, (32.42 ± 3.63)%; (0.56 ± 0.25)%, (12.14 ± 3.70)%, (19.61 ± 3.63)%, (30.43 ± 2.83)%], Bax mRNA expression level[(12.73 ± 3.96), (25.12 ± 6.42), (104.96 ± 26.77), (154.04 ± 30.52); (14.63 ± 3.57),(36.75 ± 3.67), (272.26 ± 66.11), (846.12 ± 243.36); (14.75 ± 5.65), (37.22 ± 11.27), (278.51 ± 37.42),(861.67 ± 369.29) ], Bax protein expression level [ ( 15.07 ± 0.83 ) %, ( 23.79 ± 3.99 ) %, (38.51 ± 1.58 ) %, (53.86 ±1.74)%;(15.43 ± 1.45)%,(36.11 ± 1.37)%, (56.86 ± 1.97)%, (76.09 ± 2.01)%; (15.20 ± 1.03)%,(35.25 ±1.09)%, (55.56 ± 2.17)%, (74.48 ± 2.85)% ] was respectively increased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05). The bel-2 mRNA expression level [ (443.00 ± 244.47), (156.79 ±53.18), (62.13 ± 13.66), (23.10 ± 6.44); (420.55 ± 110.77), (48.15 ± 10.02), (14.91 ± 6.53), (7.54 ± 2.62);(577.75 ± 123.22), (49.68 ± 10.11), (12.41 ± 1.28), (7.22 ± 1.89)], bcl-2 protein expression level[(47.20 ±3.77)%, (41.80 ± 2.94)%, (36.00 ± 2.36)%, (29.00 ± 2.91)%; (45.90 ± 4.15)%, (35.70 ± 2.77)%, (29.80 ±2.78)%, (24.80 ± 2.66)% ; (46.70 ± 3.47)%, (36.20 ± 2.90)%, (30.10 ± 3.21)%, (25.10 ± 2.28)% ] wasdecreased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05 ). In 3,9,15 mmol/L NaAsO2, apoptotic rate of p53 group, mRNA expression of bcl-2, protein expression of bcl-2 was higher than that ofnormal group and PC group, respectively (all P < 0.05), but mRNA expression of Bax, protein expression of Bax was respeetivelylower than that normal group and the PC group(P < 0.05 ). Conclusion p53 gene reduced the apoptosis induced by NaAsO2 in HELF, possibly by changing the apoptosis pathway.

BACKGROUNDDNA hypomethylation of long interspersed nuclear elements-1 (LINEs-1) occurs during carcinogenesis, whereas information addressing LINE-1 methylation in Wilms tumor (WT) is limited. The main purpose of our study was to quantify LINE-1 methylation levels and evaluate their relationship with relative telomere length (TL) in WT.

METHODSWe investigated LINE-1 methylation and relative TL using bisulfite-polymerase chain reaction (PCR) pyrosequencing and quantitative PCR, respectively, in 20 WT tissues, 10 normal kidney tissues and a WT cell line. Significant changes were analyzed by t-tests.

RESULTSLINE-1 methylation levels were significantly lower (P < 0.05) and relative TLs were significantly shorter (P < 0.05) in WT compared with normal kidney. There was a significant positive relationship between LINE-1 methylation and relative TL in WT (r = 0.671, P = 0.001). LINE-1 Methylation levels were significantly associated with global DNA methylation (r = 0.332, P < 0.01). In addition, relative TL was shortened and LINE-1 methylation was decreased in a WT cell line treated with the hypomethylating agent 5-aza-2'-deoxycytidine compared with untreated WT cell line.

CONCLUSIONThese results suggest that LINE-1 hypomethylation is common and may be linked to telomere shortening in WT.

Cell Line, Tumor ; Child ; Child, Preschool ; DNA Methylation ; genetics ; Female ; Humans ; Long Interspersed Nucleotide Elements ; genetics ; Male ; Polymerase Chain Reaction ; Telomere ; genetics ; Wilms Tumor ; genetics

OBJECTIVETo observe the protective effects of the Huanglian Jiedu decoction aqueous extract and its active fraction, which consists of total alkaloids, total flavonoids and total iridoid, on the thalamus of cerebral ischemia in rats.

METHODThe rat model of middle cerebral artery occlusion (MCAO) was chosen. Male SD rats were randomly divided into sham-operation group, model group, aqueous extract group (800 mg x kg(-1)), total alkaloids group(44 mg x kg(-1)), total flavonoids group (50 mg x kg(-1)) and the total iridoid group (80 mg x kg(-1)). The rats were administered the appropriate drugs intragastrically once a day, for 7 days after surgery. An equivalent volume of saline was given in the sham surgery and model groups. The HE staining was adopted to observe the pathological changes. Determination of Glu and gamma-GABA in thalamus were detected by HPLC with fluorescence detection. The expression of GAD65 was examined with immunohistochemistry and double staining with uorescent-conjugated antibodies against GFAP and Cx43 was chosen in this study.

RESULTThe neurons degenerated in MCAO rats after cerebral ischemia 7 d. The content of Glu, gamma-GABA decreased (P < 0.05), the expression of GAD65 reduced (P < 0.05) and the expression of GFAP and Cx43 increased (P < 0.01) in thalamus of rats compared with sham-operation group. Huanglian Jiedu decoction aqueous extract, total alkaloids, total flavonoids and total iridoid reduced the degeneration of neurons. Total flavonoids could promote the expression of GAD65 (P < 0.05) and decrease the expression of GFAP and Cx43 (P < 0.01) in thalamus compared with model group while it could also increased the content of Glu,gamma-GA BA to normal levels. Compared with model group, Huanglian Jiedu decoction aqueous extract, total alkaloids and total iridoid could raise the expression of Cx43, and Huanglian Jiedu decoction aqueous extract could also increase the expression of GAD65 (P < 0.05). The expression of GFAP in Huanglian Jiedu decoction aqueous extract group, total alkaloids group and total iridoid group were not different compared with model group while the content of gamma-GABA decreased (P < 0.05) compared with sham-operation group.

CONCLUSIONThe degeneration of nerve cells, the reduction of neurotransmitter amino acids content, the aberrant activation of astrocytes and the abnormal expression of GFAP and Cx43 will appear in thalamus of MCAO rats after ischemia. Huanglian Jiedu decoction total flavonoids could relieve the injury of nerve cell through inhibiting the abnormal activation of astrocytes and regulating the expression of GFAP and GAD65.

Animals ; Astrocytes ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Thalamus ; drug effects

Objective To explore and analyze the clinical values of combined therapy of colchicine and benzbromalone capsules in the treatment of gouty arthritis of non-acute episode. Methods From December 2015 to February 2017, 60 patients with gouty arthritis who were not in acute attack were randomly divided into the combined group and the control group, with 30 cases in each group. The control group was given benzbromarone capsules and the combined group was given colchicine combined with benzbromarone capsules. The changes of laboratory indexes [blood uric acid, erythrocyte sedimentation rate (ESR),C reactive protein (CRP),alanine aminotransferase (ALT), glomerular filtration rate (GFR) ], and clinical symptoms (pain, lesion joint limitation, fever, redness and tenderness) were measured and compared in the two groups before and after the treatment. The daily living ability scale (SF-36) was used to evaluate the quality of daily life before and after the treatment, and the incidence rate of drug adverse reactions during the treatment was also counted. Results No significant differences were found in the laboratory related indexes, clinical symptoms and quality of life before the treatment between the two groups (P＞0.05) . After the treatment, the levels of ALT and GFR were not significantly fluctuated in the two groups, but the levels of blood uric acid, ESR, CRP, pain symptoms and quality of life were improved, and the increasing degree in the combined group was more significant than that of the control group. and the number of episodes of acute gouty arthritis in the combined group was also lower than that in the control group, and the difference between the above indexes was statistically significant (P＜0.05) . The total incidence rate of adverse reactions was 30% in the combined group, which was not significantly different from that in the control group with 26.67% (χ2=0.180,P=0.857) . Conclusions The use of colchicine combined with benzbromalone capsules is as the safe as the single use of benzbromalone capsules and the effect of the former one is better than that of the latter one in the treatment of gouty arthritis at non-acute episode stage.

OBJECTIVETo study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress.

METHODSThe rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment.

RESULTSThe concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly.

CONCLUSIONSAt the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP.

Animals ; Blood Pressure ; Cytokines ; biosynthesis ; Hot Temperature ; Male ; Nitric Oxide ; blood ; Rabbits ; Wounds and Injuries ; blood ; physiopathology

OBJECTIVETo assess the prevalence of pre-hypertension and hypertension as well as the association with overweight/obesity in Shanghai adolescents.

METHODSSchool children aged 11 - 17 years in four schools in Shanghai were included in this survey. All students were visited in May 2010 at school by trained nurses or physicians who administered a questionnaire and carried out anthropometric measurements. Anthropometric measurements included height, weight, heart rate and blood pressure. Repeat measurements were performed in school children with elevated blood pressure within one month. The pre-hypertension and hypertension was defined on the basis of the 2004 National High Blood Pressure Education Program Working Group definitions. The overweight and obesity was defined using cutoff points recommended by Working Group of Obesity, China (WGOC).

RESULTSThere were 4175 school children aged 11 - 17 years [mean (15.0 ± 1.9) years, 52.3% (n = 2183) girls and 47.7% (n = 1992) boys]. There were 72.5% (n = 3025) participants with normal blood pressure [ girls (n = 1666) 76.3% and boys (n = 1361) 68.3%], 18.0% (n = 750)participants with pre-hypertension [14.2% (n = 310) for girls and 22.1% (n = 440) for boys], 8.3% (n = 346) participants with stage 1 hypertension [8.2% (n = 179) for girls and 8.3% (n = 165) for boys] and 1.3% (n = 54) participants with stage 2 hypertension [1.3% (n = 28) for girls and 1.3% (n = 26) for boys]. The prevalence of overweight and obesity was 6.7% (n = 147) and 3.0% (n = 66) for girls and 13.7% (n = 273) and 6.3% (n = 125) for boys. After adjusting for gender, age, parental history of hypertension and physical activities, multivariable logistic regression analysis showed that the overweight [adjusted odds ratio and 95% confidence interval: 1.42 (1.16 - 1.75)] and obesity [adjusted odds ratio and 95% confidence interval: 2.35 (1.78 - 3.11)] were independent predictors of elevated blood pressure in this cohort.

CONCLUSIONSThe prevalence of elevated blood pressure is common in adolescents in Shanghai. Overweight and obesity are closely associated with the elevated blood pressure in this cohort.

Adolescent ; Child ; China ; epidemiology ; Epidemiological Monitoring ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Obesity ; epidemiology ; Overweight ; epidemiology ; Prevalence ; Risk Factors

OBJECTIVE: To establish standardized methods and parameters of the isolated heart coronary angiography through the experiment of in vitro porcine heart by MSCT. METHODS: Based on different perfusion volume (50, 60 and 70 mL) and different perfusion-imaging time (5, 10 and 20 min), the in vitro porcine coronary artery was injected liposoluble and water-soluble contrast agents using remodel angiography equipment and scanned by MSCT. And the 3D image results were compared. The images were recorded and evaluated by 2 radiologists and analyzed by statistical software. RESULTS: Liposoluble contrast agent affected the images by damaging and infiltrating the fats around the coronary artery, while the water-soluble contrast agent didn't affect the images. The groups with 60 mL or 70 mL perfusion and 5 min perfusion-imaging time had the best images. CONCLUSION The suitable parameters of the angiography lay the foundation of postmortem coronary angiography.
Animals ; Coronary Angiography/veterinary* ; Coronary Vessels/diagnostic imaging* ; Heart ; Imaging, Three-Dimensional/methods* ; In Vitro Techniques ; Multidetector Computed Tomography/veterinary* ; Predictive Value of Tests ; Sensitivity and Specificity ; Software ; Software Validation ; Swine

OBJECTIVETo investigate the efficacy and adverse reaction of bortezomib plus chemotherapy with or without stem cell transplantation (SCT) for treatment of multiple myeloma (MM).

METHODSThirty-one MM patients were treated with bortezomib plus dexamethasone or thalidomide or DTPACE, followed by SCT. Response to bortezomib was evaluated according to the European Blood and Marrow Transplantation (EBMT) criteria. Adverse events were graded according to the WHO criteria.

RESULTS1) 5 discontinued the bortezomib therapy because of acute renal failure or acute tumor lysis syndrome and 3 died. In 26 evaluable patients received 99 courses of therapy. The overall response rate (ORR) to bortezomib was 80.8%, and was 100.0% in 15 newly diagnosed patients and 54.6% in 11 relapsed/refractory patients. All of the 6 newly diagnosed patients treated with bortezomib plus DTPACE followed by SCT achieved CR. 2) In 7 newly diagnosed patients completed 8 cycles bortezomib treatment, the diseases were improved more and more with the courses of treatment. Chromosome 13 deletion did not exert a negative impact on response. 3) 6 of 7 patients completed 8 cycles treatment without SCT relapsed in 1-3 month after discontinued therapy; only 1 of 6 such patients received SCT relapsed with the rest keeping on CR at 6-11 month follow-up. 4) The most common adverse events were 1-2 grade and tolerable 3 patients had to reduce the bortezomib dosage because of peripheral neuropathy or sinus bradycardia.

CONCLUSIONBortezomib in combination with chemotherapy with or without SCT is an effective therapy with manageable toxicities for MM patients.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Boronic Acids ; administration & dosage ; Bortezomib ; Dexamethasone ; administration & dosage ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; surgery ; therapy ; Pyrazines ; administration & dosage ; Thalidomide ; administration & dosage ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo determine the incidence and clinical significance of chromosome 13q14 deletion in multiple myeloma (MM).

METHODSBone marrow samples were collected from 132 newly diagnosed MM patients referred to our hospital. Interphase fluorescence in situ hybridization (i-FISH) combined with magnetic activated cell sorting (MACS) were performed on chromosome 13q14 (RB-1).

RESULTS(1) i-FISH was used to investigate CD138-enriched bone marrow MM cells and revealed a 13q14 deletion rate of 51.5% (68/132), while conventional cytogenetic (CC) analysis revealed 13q deletions/monosomy 13 (Δ13) only of 5.0%(6/120). (2) Univariate analysis showed that 13q14 deletion rate by i-FISH > 25%, bone marrow plasma cells > 50%, ISS stage and β(2)-MG ≥ 5.5 mg/L were associated with shorter overall survival (OS). Multivariate analysis revealed that 13q14 deletion rate by i-FISH > 25% was an independent unfavorable factor (P = 0.042). (3) Patients treated with bortezomib had a much better response than those treated with traditional chemotherapy (P = 0.001). There was no significant difference in OS between patients received bortezomib with and without 13q14 deletion (P > 0.05), indicating that bortezomib could reverse the poor prognosis of 13q14 deletion.

CONCLUSION(1) i-FISH followed CD138 cell sorting appears to be a highly sensitive method for detecting 13q14 deletion. (2) 13q14 deletion rate by i-FISH > 25% is an independent unfavorable factor. (3) Bortezomib could reverse the poor prognosis of 13q14 deletion.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Boronic Acids ; therapeutic use ; Bortezomib ; Chromosome Deletion ; Chromosome Disorders ; Chromosomes, Human, Pair 13 ; Female ; Flow Cytometry ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; drug therapy ; genetics ; pathology ; Prognosis ; Pyrazines ; therapeutic use

OBJECTIVETo verify applicability of the International Staging System (ISS) for multiple myeloma (MM) to 112 Chinese MM patients and compare ISS with Durie-Salmon (DS) and Intergroup Francophone du Myeloma (IFM) staging system in predicting prognosis.

METHODS112 previously untreated MM patients in Blood Diseases Hospital of CAMS were analyzed according to ISS retrospectively.

RESULTS1) Serum beta2-microglobulin (beta2-MG) > or = 3.5 mg/L was an independent adverse prognostic factor for overall survival (OS), and serum albumin <35 g/L predicted for time to progression (TTP), 2) In the 58 cases having cytogenetic data, chromosome 13 aberration (Delta 13) was the only independent adverse prognostic factor for OS; 3) Factors significantly related to serum beta2-MG were serum creatinine, 24h urinary protein,body mass index (BMI) and performance status (PS); and those related to serum albumin were hemoglobin level, percentage of bone marrow plasma cells, lactate dehydrogenase(LDN), fever, PS, class of M-protein, serum phosphorus and BMI; 4) All traditional prognostic factors had no statistical difference between ISS stage II and III excepting for serum beta2-MG and creatinine, and 5/6 Delta 13 patients were classified to ISS stage II; 5) The median OS of ISS stage I, II, III were 69, 23 and 26 months (m) respectively, being no statistical difference between stage II and III; for DS system, 89.5% of patients were classified in stage III, being no statistical difference for OS between the stage I/II and III; while for IFM system, the median OS of low-, intermediate- and high-risk group were 69, 40 and 8 months respectively, being statistically different between high-risk and intermediate/ low-risk groups.

CONCLUSIONSFrom the result of our limited analysis, the staging of ISS II and III seems unsuitable for Chinese MM patients. The IFM staging system ,which incorporates delta 13, is more effective than ISS, and DS staging system in predicting prognosis.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; pathology ; Neoplasm Staging ; Prognosis ; Retrospective Studies