Adolescent ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; International Cooperation ; Pediatrics ; Practice Guidelines as Topic ; standards ; Sepsis ; diagnosis ; physiopathology

Adenocarcinoma ; pathology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Nucleosides ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; Pyridazines ; administration & dosage ; pharmacology ; Stomach Neoplasms ; pathology

Objective To investigate the effect of low-dose hydrocortisone(HC)on hippocampus nuclear factor kappa B((NF-?B)),I?B expression in lipopolysaccharide(LPS)induced septic rats and the role of NF-?B signal transcription pathway in pathogenesis.Methods Fifty-four rats were randomly divided into 3 groups: control group(A group,n=6),model group(B group,n=24),low-dose HC treatment group(C group,n=24).The septic rat model was established by intraperitoneal injection LPS(1 mg/kg),as the intervention by caudal vein injection low-dose HC(6 mg/kg),each of B and C group was subdivied into 2,8,16,24 hours respectively after LPS injection(n=6).At serial time points,the animals in each group were sacrificed,brain tissue samples were harvested to determine NF-?B,I?B expression by immunhistochemistry in hippocampus.Results In B group: NF-?B expression was up regulated compared with A group(P

AIM: To study the effects of insulin and glucose on tissue-type plamingen activator (tPA) and its inhibitor-1 (PAI-1) secretion in cultured human endothelial cells. METHODS: Human endothelial cell line ECV-304 was cultured with glucose and/or insulin at different concentrations with or without hypoxic exposure. RESULTS: The tPA, PAI-1 secretion and ratio of tPA/PAI-1 increased in endothelial cells during hypoxia. Insulin and glucose increased the tPA and PAI-1 secretion in endothelial cells exposed to hypoxia, and increase in tPA/PAI-1 ratio was also observed at 4 h and 8 h. CONCLUSION: Hypoxia stimulates the release of tPA and PAI-1. Insulin and glucose also stimulate the tPA and PAI-1 secretion during hypoxia.

BACKGROUND: It has been proved that interleukin-18 (IL-18) involves in the development of acute cerebral infarction (ACI) and is positively correlated with the time of brain stroke onset, erythrocyte sedimentation rate (ESR), as well as neurological deficit and low-density value of brain CT.OBJECTIVE: To investigate the dynamic changes of serum IL-18 in the course of ACI.DESIGN: Verification analysis with patients as subjects and healthy volunteers as controls.SETTING: Department of neurology of a municipal hospital.PARTICIPANTS: Totally 46 inpatients (29 males and 17 females) with ACI were randomly selected from the Department of Neurology of People's Hospital, Maoming City, between December 2002 and January 2004.Meanwhile, 40 healthy controls (27 males and 13 females) were recruited from those who came to the hospital for routine physical examination. All the participants signed informed consent.METHODS: Fasting peripheral vein blood of 2 mL was collected from the patients on the 1st, 7rh, 14th and 21st days of ACI onset, and from healthy controls on the day of routine examination. The blood samples were centrifuged at 3 000 r/minute for 15 minutes at 4 ℃, and then the supernatant was collected for detecting IL-18 level by ELISA.MAIN OUTCOME MEASURES: The serum IL-18 level of patients on the 1st, 7th 14th and 21st days of ACI onset and that of healthy controls on the day of routine examination.RFSULTS: Totally 46 patients and 40 healthy controls were enrolled in this study and all data were statistically analyzed. The level of serum IL-18 was significantly higher in the patients with ACI than in normal controls on the 1st and 7th days of onset [(178±41) ng/L, (104±34) ng/L, (65±14) ng/L, P ＜ 0.01],but was similar on the 14th and 21st days [(88±36) ng/L, (72±33) ng/L,(65±14) ng/L, P＞ 0.05]. The level of serum IL-18 in ACI patients was significantly higher on the 1st day than on the 7th, 14th and 21 st days of the onset (P ＜ 0.05-0.01); moreover, it was also significantly higher on the 7th day than on the 21st day of the onset (P ＜ 0.05).CONCLUSION: The level of IL-18 increases obviously on the 1st day of ACI onset, and gradually decreases with the extended course of disease and time of treatment.

Objective: To objectively evaluate the short-term and long-term efficacies of arthrolysis under brachial plexus anesthesia in treating adhesive capsulitis of the shoulder (ACS). Methods: One hundred patients diagnosed with ACS were divided into two groups using the random number method. The two groups both received same active rehabilitation exercises. Besides, 55 cases in the treatment group were given one session of arthrolysis under brachial plexus anesthesia, and 45 cases in the control group were given tuina treatment. Changes in the visual analog scale (VAS) score, Melle score and pressure pain index were observed 1 month and 3 months after treatment. The therapeutic efficacies were also compared. Results: The total effective rate was 96.4％ at the 1-month follow-up and 96.4％ at the 3-month follow-up in the treatment group. The total effective rate was 33.3％ at the 1-month follow-up and 28.9％ at the 3-month follow-up in the control group. There were significant differences between the two groups comparing the total effective rate at the two time points (both P<0.05). The scores of VAS, Melle and pressure pain were significantly different at the 1-month and 3-month follow-ups from those before treatment in the treatment group (all P<0.05); the three scores did not show significant differences at the 1-month and 3-month follow-ups compared with those before treatment in the control group (all P>0.05). Conclusion: Based on the active rehabilitation exercises, one session of arthrolysis under brachial plexus anesthesia can release the adhesion and restore the range of motion and function of shoulder joint in ACS patients. It is superior to rehabilitation exercises plus tuina treatment comparing both short-term and long-term efficacies.

Adult ; Granuloma, Respiratory Tract ; microbiology ; Humans ; Male ; Mycoses ; diagnosis ; therapy ; Pharyngeal Diseases ; diagnosis ; microbiology ; therapy

OBJECTIVETo explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HuMSCs) to differentiate into Leydig cells through conditioned medium derived from Leydig cells.

METHODSHuMSCs and Leydig cells were obtained by tissue blocks culture attachment and enzymatic digestion respectively. HuMSCs were induced by conditioned medium of Leydig cells as an experiment group while those before induction were cultured as a control group. The expressions of LHR, 3β-HSD and StAR in the induced HuMSCs were determined by RT-PCR after 3, 7 and 10 days of culture; those of CYP11A1, CYP17A1 and 3β-HSD measured by immunofluorescence staining after 2 weeks; and that of 3β-HSD detected by Western blot after 4 weeks.

RESULTSThe experimental group showed positively expressed LHR, 3β-HSD and StAR at 3, 7 and 10 days, CYP11A1, CYP17A1 and 3β-HSD at 2 weeks, and 3β-HSD at 4 weeks, while the control group revealed negative expressions at all the time points.

CONCLUSIONInduced with conditioned culture medium derived from Leydig cells, HuMSCs are likely to differentiate into steroidogenic cells and eventually into Leydig cells.

Cell Differentiation ; Culture Media, Conditioned ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology

Objective To construct the bp26 deletion mutant of Brucella vaccine strain M5-90 (M5-90Δbp26),to compare its immunogenicity with parental strain(M5-90),and to develope a new candidate vaccine strain of Brucella melitensis with reduced virulence that can be used to distinguish vaccinated livestock from infected animals.Methods Mutant vaccine strain of Brucella melitensis was constructed by conventional molecular biology techniques then the genetic stability of mutant M5-90Δbp26 was tested and its conventional bacteriological nature was identified; 1.0 × 109 CFU/2 ml doses of M5-90Δbp26 strain and the parental strain were used to vaccinate 3 sheep; sera were analyzed for reactivity against BP26 by Western blotting and for agglutination activity; to analyze the virulence of mutant and parental strain,mice were injected with 1.0 × 106,6.0 × 106 and 2.0 × 107 CFU/0.2 ml doses of M5-90 and M5-90Δbp26,respectively,and clinical symptoms were monitored and the death of mice was recorded.Results The M5-90Δbp26 was successfully generated and reversion was not observed in 15 generations.The size of PCR products was 629 bp while the parental strain was 1279 bp.The sequence analysis showed a 650 bp missing in M5-90Δbp26.The conventional bacteria identification tests confirmed that the mutant was depth variant strain,including mono-specific antiserum M type transformed into R,and the BK2 phage based splitting assay converted from the positive to the negative.Western blotting showed the purified BP26 protein was recognized by the serum against the parental strain while not by the serum against M5-90Δbp26 strain.Agglutination test showed the level of the serum antibody induced by M5-90Δbp26 strain(1:50) was significantly lower than that of serum induced by parental strain(＞ 1:800).Virulence test showed that M5-90Δbp26 strain was less virulent than parental strain.Conclusions M5-90Δbp26 has been successfully constructed.M5-90Δbp26 of Brucella melitensis has the characteristic of reduced virulence and has a potential as brucellosis candidate vaccine strain permitting serological discrimination between diseased and vaccinated livestock.

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.