Objective:To explore whether microRNA-21-5p (miR-21-5p) has the effect of anti-apoptosis of human alveolar typeⅡ epithelial cells (ATⅡ).Methods:ATⅡ cells derived from the human were cultured in vitro and used for experiments when the cells were grown until the presence of lamellar bodies and microvilli were observed by light microscope. The cells were divided into blank control group (direct culture), hydrogen peroxide (H 2O 2) injury group (cultured with 0.5 mmol/L H 2O 2), and miR-21-5p overexpression group (using miR-21-5p with a multiplicity of infection (MOI) of 100 lentiviral overexpression vector with 0.5 mmol/L H 2O 2) and miR-21-5p empty virus control group (miR-21-5p lentiviral blank vector was co-cultured with 0.5 mmol/L H 2O 2). In each group, cell proliferation was detected by cell counting kit-8 (CCK-8) at 0, 12, 24, 36, and 48 hours of cell culture; cell apoptosis was detected by flow cytometry at 24 hours of culture. Results:① Cell proliferation activity test results: with the extension of cell culture time, the cell proliferation activity of the blank control group gradually increased, while the cell proliferation activity gradually decreased after the addition of 0.5 mmol/L H 2O 2. However, the cells proliferation activity in the miR-21-5p overexpression group decreased more slowly than that in the H 2O 2 injury group and the miR-21-5p empty virus control group, and the cell proliferation activity at 48 hours was significantly higher than the H 2O 2 injury group and the miR-21-5pempty virus control group ( A value: 0.295±0.005 vs. 0.184±0.005, 0.169±0.002, both P < 0.05). It showed that both H 2O 2 and lentivirus accelerated cell damage, while miR-21-5p could reduce cell apoptosis. ② Apoptosis rate test results: compared with the blank control group, the apoptosis rate increased significantly after adding 0.5 mmol/L H 2O 2; while the apoptosis rate of the miR-21-5p overexpression group was lower than that of the H 2O 2 injury group and miR-21-5p empty virus control group [early apoptosis rate: (14.31±0.12)% vs. (24.50±0.12)%, (23.41±0.13)%; late apoptosis rate: (8.12±0.13)% vs. (9.71±0.11)%, (10.41±0.15)%; overall apoptosis rate: (22.33±0.12)% vs. (34.21±0.10)%, (33.82±0.14)%; all P < 0.05], which further proved that miR-21-5p had anti-apoptotic effects. Conclusion:miR-21-5p has an anti-apoptotic effect on human ATⅡ.

Objective To determine the value of applying LIIR Automatic Analysis System of Infrared Spectroscopy in analyzing urinary stone composition. Methods 1450 samples of urinary stones were collected from 1032 male and 418 female patients. The age of patients ranged from 6 months to 88 years. The mean ages were 41.7±15.3 and 42.0±15.6 years for male and female patients, respectively. Of 1450 stones, 875 cases were located in kidney (60.34%), 504 cases in ureter (34.76%) and 71 cases in bladder (4.90%). All stones were analyzed by LIIR Automatic Analysis System of Infrared Spectroscopy (Tianjin). Analysis results were reevaluated by the artificial analysis of spectrogram, if necessary, with polarization microscope, chemical analysis, and X-ray diffraction.Results Calcium oxalate monohydrate stones were found in 714 cases (49. 24%), carbonate apatite stones in 444 cases (30.62%), anhydrous uric acid stones in 93 cases (6.41%), calcium oxalate dihydrate stones in 92 cases (6. 34 % ), ammonium magnesium phosphate hexahydrate stones in 28 cases (1.93%), cystine stones in 23 cases (1.59%), ammonium urate stones in 20 cases (1.38%), uric acid dihydrate stones in 16 cases (1.10%), brushite stones in 12 cases (0.83%), sodium urate monohydrate stones in 2 cases (0. 14%), calcium carbonate stones in 1 cases (0. 07%), and other stone types in 5 cases (0. 34%). Most urinary stones were composed of 2 or more compositions, and pure stones were only observed in 397 cases (27.38%). Most of the mixed stones contained calcium and non-calcium mixed stone was rarely observed. In addition, 15 stones were found in infants who had consumed melamine-contaminated milk powder. These stones were composed of uric acid dihydrate and ammonium urate. The results of reevaluation by artificial analysis showed the following: among pure and mixed stones, false detection occurred in 6 cases (0.41%), of which the composition was ammonium urate or carbonate apatite determined by automatic system but the true composition was anhydrous uric acid. False negative detection occurred in 9 cases (0.62%), of which the composition was ammonium magnesium phosphate hexahydrate or carbonate apatite in 7 cases, but in other 2 cases the composition could not be determined by artificial analysis. The false negative detection of components with relatively low content occurred in 6 cases and 10 cases in stones with 2 components and 3 components, respectively. The undetected composition in these cases was ammonium magnesium phosphate hexahydrate or carbonate apatite. Conclusion Automatic Analysis System of Infrared Spectroscopy has many advantages in accuracy, automation and is quick in analyzing the composition of urinary stones, and is worthy of promotion in clinical use.

Adolescent ; Colon, Ascending ; pathology ; Colonic Neoplasms ; pathology ; surgery ; Colonic Polyps ; pathology ; surgery ; Diagnosis, Differential ; Follow-Up Studies ; Ganglioneuroma ; pathology ; surgery ; Humans ; Male

OBJECTIVE： To evaluate the clinical therapeutic effect of L-glutamine granules on intestinal damage of severe burn patients and the safty of the drug.METHODS： Thirty-nine severe burn patients were randomly divided into two groups： control group（C group， nineteen patients） and L-glutamine treatment group（GLN group， twenty patients） .GLN group patients were given L-glutamine in a dose of 30g per day for 7 days， and C group patients were given the same dosage of placebo for 7 days.The plasma L-glutamine concentration， the degree of intestinal mucosa damage， blood biochemistry and complication were observed and wound healing rate of burn area was determined， then the length of hospital stay was recorded.RESULTS： After 7 days of taking L-glutamine orally， plasma L-glutamine concentration in GLN group was significant higher than that in C group（P<）. The degree of intestine damage and intestinal mucosal permeability in GLN group were lower than those in C group. In addition, the wound healing rate was faster and the length of hospital stay was shorter in GLN group than those in C group. CONCLUSION: Administration of L-glutamine could abate the degree of intestine damage obviously, lessen intestinal mucosal permeability, ameliorate wound healing rate and reduce the length of hospital stay.

Objective To investigate the effects of icarrin on the activity and protein expression of core binding factor otl(Cbfa1) in rat osteoblasts cultured in vitro,and to explore whether mitogen-activated protein kinase (MAPK) pathway is involved in this process.Methods Calvarial osteoblasts were obtained from newborn (<24 h) SD rats by trypsin-coUagenase digestion method.The second generation osteoblasts were cultured in the medium containing icariin (10 ng/ml) or estradiol (10-8 mol/L) with or without extracellular-signal regulated kinase (ERK) inhibitor (UO126) or p38MAPK inhibitor (SB203580).Nuclear protein was extracted from osteoblasts.And then the activity of Cbfa1 was detected by ELISA.The amounts of Cbfa1 protein were detected by Western blot.Results Calvarial osteoblasts were obtained successfully and were used in this study after indentified by alkaline phosphatase and mineralized nodus staining.Cbfa1 expression and the activity in osteoblasts were up-regulated by both icariin and estradiol (P<0.05).The effects were partly inhibited by addition of U0126or SB203580 (P<0.05).Conclusions Either icarrin or estradiol can stimulate the proliferation and maturation of cultured osteoblasts in vitro via up-regulating the activity and expression of Cbfal.The MAPK signal pathway inhibitor seems to partly decrease Cbfa1 activity.It suggests that MAPK pathway may be involved in the transduction of icariin's impact on proliferation and mineralization of osteoblasts.

Objective To determine the effects of β-estradiol on vasoconstriction in human umbilical artery and vein and its potential mechanisms.Methods Human umbilical cord samples were obtained from 96 term neonates of healthy singleton pregnant women born in the First Hospital of Soochow University between December 2013 and June 2015 (multiple pregnancy,pregnancy complications,cesarean delivery and low birth weight were excluded).Human umbilical arteries and veins were isolated and suspended in 37 2 organ baths containing 5 ml Krebs solution and exposed to β-estradiol followed by phenylephrine (PE) for vasoconstriction test.The subjects were divided into β-estradiol group and control group according to the presence or absence of β-estradiol incubation.To determine the effects and the possible underlying mechanisms of β-estradiol on PE-induced vasoconstriction,human umbilical artery and vein rings were pretreated with N ω-nitro-L-arginine (L-NMMA,nitric oxide synthesis inhibitor),fulvestrant (ICI182780,estradiol receptor antagonist),indomethacin (prostaglandin synthesis blocker),and removal of endothelium,then incubated with β-estradiol for 60 min followed by PE,and the concentration-response curves to PE were recorded.The concentrationresponse curves to phorbol 12,13-dibutyrate (PDBU,protein kinase C agonist) in Krebs solution in the presence or absence of β-estradiol were also obtained.Nonlinear regression and fitting curve were performed,and the two-sample ANOVA was used for analysis.Results (1) β-estradiol suppressed PE-induced vasoconstriction of human umbilical vein and artery.In human umbilical vein and artery of the control group,the maximum contraction intensity induced by PE was (59.17± 5.98)％ and (43.35± 5.02)％ of that induced by potassium chloride,respectively.The maximum contraction induced by PE in β-estradiol group was (5.87± 1.32)％and (4.52±1.22)％ of that induced by potassium chloride.(2) In both groups,incubation with L-NMMA or endothelium removal enhanced the vasoconstriction of human umbilical artery and vein,indicating that the inhibitory effect of β-estradiol was not influenced by the endothelium.(3) The suppression of β-estradiol on PE-induced vasoconstriction in human umbilical artery and vein was not significantly decreased by estrogen receptor antagonist.(4) β-estradiol did not affect human umbilical artery and vein vasoconstriction induced by PDBU.(5) In the control group,incubation with indomethacin did not affect human umbilical artery and vein vasoconstriction induced by PE.In the β-estradiol group,indomethacin significantly enhanced the contraction response induced by PE,suggesting that prostacycline synthesis was partly involved in β-estradiol-suppressed contractility in human umbilical artery and vein.The contractile response induced by phenylephrine was still lower in the β-estradiol group than in the control group,which was induced by indomethacin.Conclusions (1) β-estradiol can suppress vasoconstriction in human umbilical artery and vein,which is not dependent on endothelium and estrogen receptors,or protein kinase C activity,(2) Prostacycline synthesis is partly involved in β-estradiol-suppressed vasoconstriction in human umbilical artery and vein.

Carcinoma, Hepatocellular ; pathology ; Humans ; Leptin ; genetics ; Liver Neoplasms ; pathology ; RNA, Messenger ; genetics ; Receptors, Leptin ; genetics ; Tumor Cells, Cultured

OBJECTIVETo detect the influence of mannitol on cerebral blood flow of post-resuscitation children by transcranial Doppler (TCD).

METHODSThe blood flow changes of left side middle cerebral artery (MCA) and extracranial internal carotid artery (EICA) were monitored by TCD in 21 post-resuscitation children. TCD waveforms, peak velocity (Vp), diastolic velocity (Vd), mean velocity (Vm) and pulsitility index (PI) of MCA and EICA were monitored daily and 30 minutes before and after the first dose of mannitol. Glasgow scores were estimated in the meantime. The patients were divided into 2 groups according to the patterns of diastolic flow. Patients in group I showed no diastolic flow or retrograde diastolic flow (n = 9), and patients in group II had positive diastolic flow (n = 12).

RESULTSVp, Vd, Vm, PI of MCA and EICA in group I patients had no significant change after the administration of mannitol and the Glasgow scores were much lower than that of group II patients (P < 0.05). TCD waveforms showed no improvement in group I patients and their outcomes were poor. Vd, Vm and Vp of MCA and Vd of EICA in group II patients increased, PI of MCA decreased (P < 0.05), while Vp, Vm, PI of EICA had no significant change. TCD waveforms recovered within one week. All the patients in this group survived.

CONCLUSIONSThe results suggested that mannitol could improve cerebral perfusion in patients with mild brain damage. These patients usually had increased diastolic blood flow in the early stage. Lack of or retrograde diastolic flow of TCD waveforms might be associated with severe brain damage with poor outcome. Mannitol had no effect on improving cerebral blood flow in these patients.

Adolescent ; Blood Flow Velocity ; Cardiopulmonary Resuscitation ; Cerebrovascular Circulation ; drug effects ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Mannitol ; pharmacology ; Middle Cerebral Artery ; physiopathology ; Ultrasonography, Doppler, Transcranial ; methods

OBJECTIVETo develop a draft questionnaire (China Musculoskeletal Questionnaire, CMQ) for evaluating of musculoskeletal workload and associated potential hazardous working conditions as well as musculoskeletal symptoms of workers in Sitting Posture.

METHODSMulti-methods, which include the reviewing references, the summarizing results of preliminary studies, the reviewing ergonomic tools, the consulting experts and occupational health workers and the interviewing or discussing with individual workers in sitting posture, were used in developing item pool. The experts and epidemiologists of occupational health scored the importance of every single item in the item pool, and then the survey and sampling were carried out in 325 workers of sitting posture who completed the questionnaire. On the basis of these data, the methods including experts scoring, item analysis, Cronbach's α analysis and factor analysis were synthetically used to select the reliable items which consisted of the formal questionnaire.

RESULTSThe standard of the CMQ, which consists of 34 items on musculoskeletal workload and associated potentially hazardous working conditions, can be divided into nine indices (dynamic loads, static loads, repetitive loads, forces-exertion, prolong time, climatic factors, vibration, position and ergonomic environmental factors).

CONCLUSIONThe CMQ possesses good content validity, and the items of CMQ are divergent, reliable and typical. However, the reliability and validity of CMQ should be validated.

China ; Ergonomics ; Humans ; Musculoskeletal System ; Occupational Health ; Posture ; Surveys and Questionnaires ; Workload

BACKGROUNDFew studies have examined the properties of human immunodeficiency virus type 1 (HIV-1) epitope-specific cytotoxic T lymphocyte (CTL) responses in children. To address this issue, we characterized epitope-specific CTL responses and analyzed the determinants that may affect CTL responses before and after highly active antiretroviral therapy (HAART) in children with HIV-1 infection.

METHODSA total of 22 HIV-1-infected children and 23 uninfected healthy children as control were enrolled in the study. Circulating CD4 T cells and HIV-1 RNA load in plasma were routinely measured. Peripheral HIV-1-specific CTL frequency and HIV-1 epitope-specific, interferon-gamma (IFN-gamma)-producing T lymphocytes were measured using tetramer staining and enzyme-linked immunospot (ELISPOT) assay, respectively. Circulating dendritic cell (DC) subsets were monitored with FACS analysis.

RESULTSMore than 80% of the children with HIV-1 infection exhibited a positive HIV-1-epitope-specific CTL response at baseline, but HIV-specific CTLs and IFN-gamma-producing lymphocytes decreased in patients who responded to HAART in comparison with non-responders and HAART-naive children. The duration of virus suppression resulted from HAART was inversely correlated with CTL frequency. While in HAART-naive children, HIV-1-specific CTL frequency was positively correlated with myeloid DC (mDC) frequency, although the cause and effect relationship between the DCs and CTLs remains unknown.

CONCLUSIONSHIV-1-epitope-specific CTL responses are dependent on antigenic stimulation. The impaired DC subsets in blood might result in a defect in DC-mediated T cell responses. These findings may provide insight into understanding the factors and related mechanisms that influence the outcome of HIV-1 carriers to HAART or future antiviral therapies.

Adolescent ; Antiretroviral Therapy, Highly Active ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Epitopes ; immunology ; Female ; HIV Infections ; drug therapy ; immunology ; HIV-1 ; immunology ; Humans ; Male ; T-Lymphocytes, Cytotoxic ; immunology ; Viremia ; drug therapy