Objective To study the effects of chloroquine on proliferation and cell cycle of hepatocellular carcinoma cell line HepG2 cultured in vitro and provide basis for research on chloroquine in therapy for liver cancer.Methods HepG2 cells were divided into six groups:control group and chloroquine (8.00,16.00,32.00,64.00 and 128.00 mmol?L-1 )groups.The cell viability was determined by MTT,the cell cycle and apoptosis were detected by flow cytometry.Results Compared with control group,the cell viabilities were inhibited significantly 24 h after treated with chloroquine (32.00-128.00 mmol?L-1) or 48-72 h after treated with chloroquine (8.00-128.00 mmol?L-1) ( P

Objectlve investigate the role of Toll-like receptor 2 (TLR2) on polymorphonuclear neutrophil (PMN) during perioperative period in the development of postoperative systemic inflammatory response syndrome (SIRS) in patients undergoing orthotopic liver transplantation (OLT).Methods Twenty patients (18 male and 2 female, aged 33-58 yr and weighing 52-73 kg) with ASA Ⅲ or Ⅳ (NYHA Ⅱ or Ⅲ )undergoing OLT were studied. Blood samples were collected from the central vein for determination of TLR2 expression on PMN and plasma TNF-α, IL-1β and IL-8 concentrations before induction of anesthesia (T1, baseline), at 25 min of anhepatic phase (T2), 3 h (T3) and 24 h after beginning of reperfusion of the allograft (T4). The expression of TLR2 was measured by flow cytometry and the serum concentrations of TNF-α, IL-1β and IL-8 were measured by enzyme linked immunosorbant assay (ELISA). The patients were divided into SIRS and non-SIRS group depending on whether the patients developed SIRS or not within 7 days after operation. The diagnosis of SIRS was based on the criteria laid down by ACCP and SCCM in 1992.Results Ten patients developed SIRS within 7 days after operation. There was no significant difference in Child-Turcotte-Pugh (CTP) scores between the two groups. Compared with non-SIRS group, the TLR2 expression on PMN and the serum IL-1β concentration were significantly increased at T4 and the serum IL-8 concentration was significantly increased at T3 in SIRS group.There was positive correlation between serum TNF-α concentration and TLR2 expression on PMN in SIRS group ( r= 0.607, P ＜0.05).Conclusion The expression of TLR2 on PMN increases significantly at 24 h after beginning of reperfusion of allograft and may play an important role in the development of postoperative SIRS.

AIMTo investigate the effects of hypoxia on the proliferation and apoptosis of PASMC, to evaluate the role of iNOS protein expression and ADM on the hypoxic pulmonary hypertension (HPH) pathogenesis.

METHODSTo culture rat pulmonary artery smooth muscle cell (PASMC), cultured PASMC cells were grouped into: normoxic group; hypoxic group; hypoxia + L-NAME group; hypoxia+ ADM group. Proliferation of PASMC were investigated by MTT and PCNA. Apoptosis of PASMC were examined by flow-cytometry. Westen blot was used to measure protein expression of iNOS induced by hypoxia.

RESULTS(By MTT, the value of 24 h hypoxia was significantly higher than that in the normoxic group (P < 0.01), the value of the hypoxia + ADM was significantly lower than that in hypoxia group, the value of the hypoxia + L-NAME was significantly higher than those of hypoxic group and normoxic group (P < 0.01). (2) By immunohistochemistry, PCNA was poorly positive in PASMC, whereas positive after 24 h hypoxia (P < 0.01), ADM inhibited the expression of PCNA significantly (P < 0.01), whereas L-NAME increased the expression of PCNA significantly (P < 0.01). (3) By FCM, apoptosis index was not significantly different between the normoxic group, hypoxic group, hypoxia + L-NAME and hypoxia + ADM (P > 0.05). (4) By Western blot, iNOS expression was poorly positive in control group, positive after 4 h hypoxia (P < 0.01), increasing as the hypoxia environment continued (P < 0.01). L-NAME had no effect on iNOS protein, ADM promoted iNOS expression (P < 0.01).

CONCLUSION(1) Hypoxia stimulates the proliferation of PASMC, and has no obvious effects on the apoptosis of PASMC. (2) Hypoxia induces the expression of iNOS, ADM can increase expression of iNOS, ADM and INOS plays a role of protection in HPH pathogenesis.

Adrenomedullin ; pharmacology ; Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Proliferation ; drug effects ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; pathology ; Nitric Oxide Synthase Type II ; metabolism ; Pulmonary Artery ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the neuroprotective effects of the constituents in Herba Erigerontis on neuroblastoma SH-SY5Y cells, and find out its possible material foundation in treating Alzheimer's disease (AD).

METHODDifferent combinations of the three constituents in Herba Erigerontis were prepared according to the orthogonality experiment, and the indexes (MTT reduction assay, lipid peroxidation and expressions of nAChR alpha7 protein)were observed upon the SH-SY5Y cells followed by treatment of these combinations and beta-amyloid peptide (AP). The pharmacology data thus obtained and peak data in UPLC fingerprint were analyzed through ANOVA and correlationship by SPSS to give the information of active possible material foundation.

RESULTConstituents B and C showed clear activity and peaks of 4, 7-12 did positive correlationship according to the correlation of fingerprints and pharmacology.

CONCLUSIONThis study makes a valid approach for deducing the active constituents even the exact compounds against neurotoxicity induced by Abeta by correlation of fingerprints and pharmacology.

Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; toxicity ; Analysis of Variance ; Animals ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Erigeron ; chemistry ; Humans ; Nervous System ; drug effects ; Neurotoxins ; toxicity

Objective To explore the correlation and diagnostic value of serum visfatin (visfatin),high sensitive C reactive protein (hs-CRP) and tumor necrosis factor-α (TNF-α) in patients with different types of angina pectoris.Methods Eighty-four patients with angina pectoris who were treated in the Eighth hospital of Wuhan from January 2013 to December 2016 were selected as the research subjects,the patients were divided into stable angina pectoris (SAP) group with 39 cases and unstable angina pectoris (UAP) group with 45 cases according to the classification criteria of angina pectoris,Thirty-seven healthy volunteers in the outpatient department of the same period were selected as the control group.To compare the levels of serum visfatin,hs-CRP and TNF-alpha in three groups,analyze the correlation between the levels of serum visfatin and hs-CRP and TNF-alpha levels in patients with angina pectoris,and analyze the diagnostic value of the levels of serum visfatin,hs-CRP,TNF-α and plasma oxidized low density lipoprotein (Ox-LDL) by ROC curve.Results There were no significant differences in fasting blood glucose,uric acid,creatinine,total cholesterol (TC),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C) among the three groups (all P＞0.05).The systolic pressure (SBP) was (134.27± 11.96) mmHg and diastolic pressure (DBP) was (84.23 ± 7.15) mmHg in SAP group,the SBP was (135.98±12.33) mmHg and DBP was (84.47±7.22) mmHg in UAP group,which were all higher than SBP ((121.49±9.97)mmHg)and DBP ((77.38±6.94)mmHg) in the control group (all P＜0.05),but there was no significant differences between the SAP group and the UAP group (all P＞0.05).The level of visfatin in UAP group was (6.57± 1.36) μg/L,which was higher than that in SAP group ((4.62 ± 1.08) μg/L) and in control group((4.27± 1.03)μg/L) (all P＜0.05),but there was no significant differences between the SAP group and the control group (P＞0.05).The level of serum hs-CRP,TNF-α and plasma Ox-LDL were (12.38±2.63) mg/L,(4.35±0.28) ng/L and (617.32±53.48) μg/L in UAP group,and the level of serum hs-CRP,TNF-α and plasma Ox-LDL were (8.29 ± 1.74) mg/L,(1.82 ± 0.34) ng/L and (418.57±43.85) μg/L in SAP group,which were higher than hs-CRP (4.06±1.52) mg/L,TNF-α (0.98 ±0.13) ng/L,Ox-LDL (213.64±32.83) μg/L in control group(all P＜0.05),and the UAP group was higher than that in the SAP group (all P＜0.05).The level of serum visfatin in group SAP and group UAP had no significant correlation with age,body mass index,SBP,DBP,fasting blood glucose,uric acid,creatinine,total cholesterol,triglyceride,HDL cholesterol,low density lipoprotein cholesterol (all P ＞ 0.05).The serum visfatin level was positively correlated with serum hs-CRP,TNF-alpha level and plasma Ox-LDL level in SAP group.(r=0.687,0.634,0.656,all P＜0.05).The serum visfatin level was positively correlated with the serum hs-CRP,TNF-a level and Ox-LDL level in UAP group (r =0.721,0.652,0.673,all P＜ 0.05).ROC curve analysis showed that the levels of serum visfatin,hs-CRP,TNF-α and plasma Ox-LDL were all near 0.7,and the sensitivity and specificity were all above 0.6 identification of different types of angina pectoris.Conclusion The level of serum visfatin in patients with UAP was significantly higher than that of SAP patients,its level is related to serum hs-CRP,TNF-alpha and plasma Ox-LDL levels,the four indexes have a certain predictive value for different types of angina pectoris,and can provide reference for different types of angina.

Object To verify and evaluate the performances of Sysmex XN-9000 hematology analyzer under body fluid mode when used for peritoneal and pleural fluids detection.Methods According to the guidance of ICSH, Sysmex XN-9000 hematology analyzer under body fluid mode had its performances verified on background counting, carryover, precision, linear range and etc of RBC-BF and TC-BF, and then was compared with microscopy on RBC-BF, TC-BF, PMN％, MN％, N％, L％, M％and E％.Results The RBC-BF and TC-BF had the background counting being 0.00, carryover being 0.00％and 0.07％ respectively, coefficients of variation (CV) of within-run precision being from 0.96％to 17.89％as well as from 2.19％to 10.33％respectively, CV of between-run precision being from 3.34％to 4.73％as well as from 8.33％to 10.75％resptctively, and the linear ranges being (0～5) ×1012/L and (0～20) ×109/L respectively. There was a high correlation between Sysmex XN-9000 hematology analyzer and microscopy when detecting RBC-BF, TC-BF, PMN％, MN％, N％, L％, M％and E％. The Spearman rank correlation coefficients (rs) were 0.977, 0.995, 0.863, 0.929, 0.926, 0.949, 0.965 and 0.816 (P<0.05), and there were statistical significances between the correlations. Bland-Altman bias analysis on the analyzer and microscopy showed that RBC-BF had the bias (4.6％) lower than that (5.6％) of TC-BF;MN％had the bias (-2.0％) lower than that (4.2％) of PMN％;L％, N％, M％and E％had the bias being-0.5％, 4.4％,-5.9％and-1.6％respectively, which were all met the requirements of the manufacturer.Conclusion Sysmex XN-9000 hematology analyzer under body fluid mode proves its performances for routine detection of peritoneal and pleural fluids.

Objective: To assess the trend of lifestyle changing instruction for acute myocardial infarction (AMI) patients at discharge in China from 2001-2011. Methods: Based on national representative data of China PEACE retrospective AMI study and the number of instructions AMI patients received at discharge, our research was classified into 3 groups: no instruction group, (1-2) instructions group and (3-5) instructions group. We described the instruction rates in different groups with 10-year trend and no instruction status in 5 regions of eastern urban, central/western urban, eastern rural, central rural, and western rural. In addition, we reported the instruction rates of diet consulting, exercise consulting, blood lipid evaluation, smoking cessation, and weight control with 10-year trend. Results: A total of 16100 medical records of AMI from 162 hospitals were enrolled. No instruction rates at discharge were 76.6%, 65.7% and 54.4% in 2001, 2006 and 2011, respectively, Ptrend<0.001. The proportion for receiving (1-2) instruction(s) at discharge increased from 23.2% in 2001 to 42.9% in 2011, Ptrend<0.001; the rate for receiving (3-5) instructions was 2.7% in 2011.No instruction rates in 5 regions were from 47.1% to 69.5%; no instruction rates in central and western rural regions had little changes during the 10-year period, Ptrend=0.11 and Ptrend=0.10, respectively; no instruction rates in the rest 3 regions showed decreasing trend, all Ptrend<0.001. Most of the single instruction rates for lifestyle changing increased from 2001 to 2011, diet consulting were 16.6%, 27.8%, and 38.3% respectively, Ptrend<0.001; exercise consulting were 9.2%, 10.9% and 14.3% respectively, Ptrend<0.001; from 2001 to 2011, blood lipids evaluation was increased from 0.9% to 11.9%; smoking cessation increased from 2.2% to 8.8%, both Ptrend<0.001; however the weight control rate was 1.3% in 2011, which was had little change within 10 years, Ptrend=0.32.

OBJECTIVETuberculosis is still a public health problem. Host genetic factors, such as polymorphisms in NRAMP1 gene, may play a role in the development of tuberculosis. To clarify the effect of NRAMP1 gene polymorphisms on the development of childhood tuberculosis, the association of NRAMP1 gene polymorphisms with susceptibility to tuberculosis in the ethnic Han Chinese children was investigated.

METHODSFrom January 2005 to March 2008, 130 ethnic Han children with tuberculosis (TB group) were enrolled. Three hundred and ninety hospitalized ethnic Han children for physical examination in the surgery department were used as the control group. The controls were matched with tuberculosis children by age, sex and area. PCR-RFLP analysis was performed on DNA samples to identify allele genotypes of INT4 and D543N in NRAMP1 gene. Genotype frequency differences between tuberculosis patients and controls were analyzed using x2 test.

RESULTSNo statistical difference was found in the genotype frequency of variants G/C and C/C at the INT4 locus between the TB and the control groups. At the D543N locus, the frequency of genotype variants (G/A and A/A) was significantly higher in the TB group (34/130) than that in the control group (66/390) (x2=5.349, P<0.05; OR=1.74, 95%CI=1.08-2.79). When stratified by sex, differences in the genotype distribution were observed only in females at the D543N locus, which the variant genotypes were higher in the TB group (16/52) than in the control group (21/155) (x2=7.866, P<0.05; OR=2.84, 95%CI=1.34-5.99). For males, there was no difference between the TB and the control groups. At the INT4 locus, no difference was observed between the two groups in boys and girls.

CONCLUSIONSGenotypic variation at the D543N locus in NRAMP1 gene may be associated with susceptibility to tuberculosis in ethnic Han Chinese children. Variant genotypes in NRAMP1 gene (G/A and A/A) may be susceptible genotypes to tuberculosis in ethnic Han Chinese children. Girls with variant genotypes were more susceptible to tuberculosis.

Cation Transport Proteins ; genetics ; Child ; Child, Preschool ; China ; ethnology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Male ; Polymorphism, Genetic ; Sex Factors ; Tuberculosis ; ethnology ; genetics

Objective To investigate the effects of regular and irregular mandatory treadmill running (TR) on learning and memory abilities,stress response,survival and maturation of newborn neurons in dentate area of C57 mice. Methods Forty-eight male C57BL/6 mice were randomly divided into regular running group (n=16,definite time and quantity), irregular running group (n=16,definite time and different quantities) and sedentary group (n=16).Injection of bromodeoxyuridine (BrdU,50 mg/kg) was given intraperitoneally to mice from the 1st d to the 7th d of exercise,and then,these mice were subjected to a 4-wk TR training course or sedentary exercise.Double irnmunofluorescence labeling was employed to observe the effect of TR on survival and maturation of newborn neurons. Mice were trained on the Morris water maze to test the abilities of learning and memory. Corticosterone (CORT).level in the venous blood was measured with a radioimmunoassay kit. Results As compared with that in mice of the irregular running group and sedentary group,significant decrease of latency was noted in mice of the regular running group on the 1st, 3rd, 4th and 5th d of place navigation test (P＜0.05). As compared with that in mice of the sedentary group, significant decrease of latency was noted in mice of the irregular running group and regular running group on the 2nd d of place navigation test (P＜0.05),while no significant difference of that was noted between mice of the irregular running group and regular running group (P＞0.05).The platform was removed for a 60-s probe test 4 h after the last trial to test the recall ability, and the times of staying in the target zone in each group were regular running group＞irregular running group＞control group, and significant difference was noted between each 2 groups (P＜0.05). The numbers of double immunofluorescence labeled cells in the dentate gyrus were counted as regular running group＞irregular running group＞control group, and significant difference was noted between each 2 groups (P＜0.05). The serum CORT level was measured as regular running group＜irregular running group＜control group, and significant difference was noted between each 2 groups (P＜0.05). Conclusion Mandatory treadmill running could improve the learning and memory abilities,which may be related to the increment of survival and maturation of neural precursor cells,and decreased level of serum CORT; and the effect of regular mandatory treadmill running is better than that of irregular mandatory treadmill running.

OBJECTIVETo evaluate the clinical efficacy of mobilized autologous peripheral blood stem cells (PBSC) transplantation in a 48 years old patient with lower limb arteriosclerosis obliterans (ASO).

METHODSrhG-CSF 600 micro g/d for 5 days to mobilize stem cells. On the fifth day, PBSC were collected with a Version 4 blood-cells separator. Three hours late, the PBSC were intramuscularly injected into the ischemic areas of the two lower limbs (3 x 10(9) cells per limb). The clinical and laboratory findings were monitored every week for 3 months. Forty-four days after the implantation, left lower limb with severe ASO was given an additional implantation of the same number of cells as the first time.

RESULTSThe peripheral blood CD(34)(+) cells were increased from 0.18% to 0.75% after 5 days of rhG-CSF mobilization. Three months after the first stem cell transplantation, severe pain lameness, local cool-feeling and ulcer were improved, and ABI increased from 0.49, 0.69 to 0.50, 0.85, the amplitude of blood flow and laser Doppler blood perfusion were also significantly improved (P < 0.01). At the same time, digital subtraction angiographic scores for new collateral vessel formation were showed as + 3(rich). No related complication or adverse effect were observed during the 3-month observation.

CONCLUSIONTransplantation of mobilized autologous PBSC might be a simple, safe, and effective method for the treatment of ASO.

Arteriosclerosis Obliterans ; therapy ; Humans ; Lower Extremity ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous