OBJECTIVE To develop a rapid system for bacteria identification and susceptibility test in 24 hours,and provide bacteriological evidence for the control of nosocomial infection and the timely diagnosis and treatment.METHODS The simple constant temperature box was replaced by revolving constant temperature box;2,3,5-triphenyltetrazolium chloride(TTC) and succinate were added to the culture bottle and the culture medium of drug susceptibility;the concentration of the reactive substrate in the bacterial biochemical tube and the number of the inoculated bacteria were increased at the same time.RESULTS The time of positive blood culture in the revolving constant temperature box was significantly shorter than that in the simple one(?2=74.92,P

The aim of this study was to explore the effect of different conditions on two-dimensional gel electrophoresis of proteins from human acute promyelocytic leukemia cell line NB4. The 24 cm pH 3-10 linear immobilized pH gradient (IPG) strips were chosen, the isoelectric focusing was carried out by using IPGphor. Then, the second-dimensional SDS-PAGE was performed. After silver staining, the gel was analyzed by ImageMaster 2D Elite. The results showed that low ion intensity sample washing buffer improved the performance of isoelectric focusing. The lysis buffer containing 7 mol/L urea and 40 mmol/L DTT could solubilize the most proteins from NB4 cell line. The rehydration solution containing thiourea and urea increased the low molecular weight protein points to be resolved in the area of basic end. The reasonable sample load and Volt/hour of NB4 were about 100 micro g and 63 200 V/h for the 24 cm pH 3-10 IPG strips. It is concluded that the proteins from NB4 and similar cell line are complicated and affected by many factors, so that, it is very important to select the right methods for sample preparation and the conditions of two-dimensional gel electrophoresis.
Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Isoelectric Focusing ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Neoplasm Proteins ; analysis

OBJECTIVETo analyze the frequency distribution of single nucleotide polymorphisms (SNPs) of four asthma-related gene loci (ACE I/D; ADRB2 Arg16Gly; TNF-α G-308A; MS4A2 Glu237Gly) in 198 asthmatic children, and to investigate its association with genetic susceptibility to childhood asthma and some clinical phenotypes of asthma.

METHODSPolymerase chain reaction product electrophoresis identification and real-time quantitative PCR detecting system were used to determine the frequency distributions of the SNPs of the four asthma-related gene loci in 198 asthmatic children and 110 healthy controls. The serum total IgE (TIgE) levels and blood eosinophil proportion (%EOS) of the asthmatic children were measured. Different genotypes at the four asthma-related gene loci were compared in terms of TIgE and %EOS.

RESULTSThe genotype DD of angiotensin-converting enzyme (ACE) had a significantly higher frequency in the asthmatic children than in the healthy controls (χ2= 30.667, P<0.01), and the frequency of D allele was also significantly higher in the asthmatic children than in the healthy controls (χ2=7.151, P<0.01). No correlation was found between the polymorphism of each gene locus and serum TIgE level and %EOS (P>0.05).

CONCLUSIONSGenotype DD of ACE is related to genetic susceptibility to childhood asthma and may be the risk factor for childhood asthma.

Asthma ; etiology ; genetics ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, beta-2 ; genetics ; Receptors, IgE ; genetics ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVESepsis and septic shock remain a common problem that results in significant mortality and morbidity in pediatric intensive care units (PICU). According to literature, the use of more physiologic steroid replacement therapy is associated with hemodynamic and survival benefits in adult patients with relative adrenal insufficiency (RAI) and catecholamine-resistant septic shock. But little information is available in children. The aim of the current prospective study was to determine the prevalence of adrenal insufficiency in children with sepsis and septic shock using a low-dose adrenocorticotropic hormone (ACTH) stimulation test (1 microg/1.73 m2) in children.

METHODSThe authors performed cortisol estimation at baseline and after low-dose (1 microg/1.73 m2) ACTH stimulation at 30 mins in children during the first 24 hours in patients with sepsis or septic shock admitted to our PICU. Adrenal insufficiency was defined as a response < or = 90 microg/L. Absolute adrenal insufficiency (AAI) was further defined as baseline cortisol (T0) < 200 microg/L and RAI insufficiency by T0 > or = 200 microg/L.

RESULTSSixty-two consecutive cases with sepsis and septic shock admitted to PICU of Shanghai Jiaotong University Affiliated Children's Hospital from April, 2006 to March, 2007. The median age was 37.6 months (range, 2 - 168 months), and their gender distribution was 42 (67.7%) males and 20 (32.3%) females, 53 cases had sepsis (85.5%) and 9 had septic shock (14.5%). The mean pediatric critical illness score (PCIS) was 79.3 +/- 9.2 and median pediatric risk of mortality score (PRMSIII) 11.3 (5 - 19), respectively. Overall mortality of sepsis and septic shock was 27.42%. The evaluation of adrenal insufficiency was conducted as follows. (1) The mean cortisol levels at baseline (T0) and 30 mins after ACTH stimulation (T1) were (318.6 +/- 230.4) microg/L, (452.3 +/- 230.7) microg/L and (454.7 +/- 212.7) microg/L, (579.3 +/- 231.9) microg/L in patients with severe sepsis and septic shock group, respectively. There were no significant difference between the two groups (P > 0.05). (2) The proportion of patients with adrenal insufficiency in the study population was 40.3% as defined by a response < or = 90 microg/L post test. The proportion of patients with adrenal insufficiency in sepsis and septic shock were 39.6% and 44.4%, respectively (chi2) = 0.073, P > 0.05). (3) The serum T0 and T1 levels were (320.5 +/- 223.9) microg/L, (462.3 +/- 212.0) microg/L and (384.3 +/- 258.3) microg/L, (500.7 +/- 470.6) microg/L, respectively, and the proportion of patients with adrenal insufficiency were 37.8% and 47.1% in the survivors and the dead (P > 0.05). The levels of T0 and T1 were related to the PCIS (P < 0.05). The morbidity of adrenal insufficiency was not related to the PCIS, PRISMIII, and number of organ that developed functional insufficiency (P > 0.05).

CONCLUSIONSAdrenal insufficiency may occur in patients with sepsis and septic shock in children. ACTH stimulation test may be helpful to determine whether corticosteroid therapy has a survival benefit in patients with relative adrenal insufficiency. A low-dose ACTH stimulation test can be used to evaluate the adrenal function status of severe sepsis and septic shock in children.

Adolescent ; Adrenal Insufficiency ; diagnosis ; etiology ; Adrenocorticotropic Hormone ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Intensive Care Units, Pediatric ; Male ; Prospective Studies ; Sepsis ; physiopathology ; Shock, Septic ; physiopathology ; Systemic Inflammatory Response Syndrome ; physiopathology

OBJECTIVETo investigate the potential effects of arsenic trioxide (As(2)O(3)) combined with 8-(4-chlorophenylthio) adenosine 3', 5'-cyclic monophosphate (8-CPT-cAMP) on the retinoic acid (RA)-resistant acute promyelocytic leukemia (APL) cells.

METHODSThe RA resistant APL cell lines NB4-R1 and NB4-R2 were used as in vitro models. The effect of As(2)O(3) and/or 8-CPT-cAMP was evaluated according to cellular morphology, cell surface antigen and nitroblue-tetrazolium (NBT) assay. Meanwhile, immunofluorescence analysis and Western blot assay were used to detect the degradation of PML-RAR alpha fusion protein and the change of several key cell cycle regulatory proteins in these cells before and after the treatment.

RESULTSLow dose of As(2)O(3) (0.25 micromol/L) synergized with 8-CPT-cAMP (200 micromol/L) in inducing differentiation of NB4-R1 and NB4-R2 cells, while neither of these two drugs alone could induce differentiation of these cells. In addition, 8-CPT-cAMP was able to inhibit the cell growth by modulating the expression of some important cell cycle regulators and to facilitate the As(2)O(3)-mediated degradation of PML-RAR alpha fusion protein.

CONCLUSIONSAs(2)O(3) combined with 8-CPT-cAMP could induce differentiation of RA-resistant APL cells.

Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cyclic AMP ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Oxides ; pharmacology ; Thionucleotides ; pharmacology ; Tretinoin ; pharmacology

Calcifying tendinopathy is a tendon disorder with calcium deposits in the mid-substance presented with chronic activity-related pain, tenderness, local edema and various degrees of incapacitation. Most of current treatments are neither effective nor evidence-based because its underlying pathogenesis is poorly understood and treatment is usually symptomatic. Understanding the pathogenesis of calcifying tendinopathy is essential for its effective evidence-based management. One of the key histopathological features of calcifying tendinopathy is the presence of chondrocyte phenotype which surrounds the calcific deposits, suggesting that the formation of calcific deposits was cell-mediated. Although the origin of cells participating in the formation of chondrocyte phenotype and ossification is still unknown, many evidences have suggested that erroneous tendon cell differentiation is involved in the process. Recent studies have shown the presence of stem cells with self-renewal and multi-differentiation potential in human, horse, mouse and rat tendon tissues. We hypothesized that the erroneous differentiation of tendon-derived stem cells (TDSCs) to chondrocytes or osteoblasts leads to chondrometaplasia and ossification and hence weaker tendon, failed healing and pain, in calcifying tendinopathy. We present a hypothetical model on the pathogenesis and evidences to support this hypothesis. Understanding the key role of TDSCs in the pathogenesis of calcifying tendinopathy and the mechanisms contributing to their erroneous differentiation would provide new opportunities for the management of calcifying tendinopathy. The re-direction of the differentiation of resident TDSCs to tenogenic or supplementation of MSCs programmed for tenogenic differentiation may be enticing targets for the management of calcifying tendinopathy in the future.
Animals ; Cell Differentiation ; physiology ; Humans ; Mice ; Rats ; Stem Cells ; pathology ; Tendinopathy ; etiology ; pathology ; Tendons ; pathology

Instruction is the basis for the clinical use of drugs and is the safe and effective legal basis for patient to protect their use of drugs. However, the dosage and usage of the instruction in the clinical practice can not meet the clinical needs, which lead to the excessive usage.In this paper, expert consensus on the excessive use of antimicrobial agents beyond the instruction is interpreted in order to provide reference for clinical medication.

Objective To demonstrate the nuclear orphan receptor Nur77 agonist fungal polyketone (Cytosporone B,Csn-B) protects the neurons in Parkinson's disease cell model.Methods The SH-SY5Y cells were divided into blank group,control group (Csn-B),model group (MPP +) and experimental group (MPP + + Csn-B).The relative viability of human neuroblastoma cells SH-SY5Y induced by 1-methyl-4-pehny1-pyridine (MPP+) was measured with MTT assay.The expression of microtubule associated protein 1 light chain3 (LC3)-Ⅱ / Ⅰ and α-synuclein protein was detected by Western blotting.The expression of LC3 mRNA and α-synuclein mRNA were detected by qPCR.Immunofluorescence was used to observe the expression of LC3 and α-synuclein in each group.Results The MTT results showed that MPP + significantly inhibited proliferation of SH-SY5Y cells in a dose dependent manner.Western blotting showed that,LC3-Ⅱ and LC3-Ⅰ gray scale in blank group,control group,model group and experimental group were 0.21 ±0.02,0.23 ±0.03,0.32 ±0.03,0.69 ±0.05,LC3-Ⅱ/Ⅰ in model group and experimental group was significantly higher than that in blank group(P ＜ 0.01),and LC3-Ⅱ/Ⅰ in experimental group was significantly higher than that in model group (P ＜ 0.01).The α-synuclein in blank group,control group,model group and experimental group were 0.30 ± 0.05,0.51 ± 0.06,2.18 ± 0.12,0.73 ± 0.08,the expression of α-synuclein in model group and experimental group were significantly higher than that in blank group (P ＜0.01).The expression of α-synuclein protein in experimental group was significantly lower than that in model group (P ＜ 0.01).The relative expression of LC3 mRNA in blank group,control group,model group and experimental group were 3.94 ±0.18,4.04 ±0.13,6.01 ±0.21,7.43 ±0.35,the LC3 mRNA in model group and experimental group were significantly higher than that in blank group (P ＜ 0.01),LC3 mRNA in experimental group was significantly higher than that in model group (P ＜0.01).The α-synuclein mRNA in blank group,control group,model group and experimental group were 3.09 ±0.19,3.47 ±0.24,7.18 ±0.30,4.60 ±0.27,the expression of α-synuclein mRNA in model group and experimental group were significantly higher than that in blank group (P ＜0.01).The expression of α-synuclein mRNA in experimental group was significantly lower than that in model group (P ＜0.01).Immunofluorescence showed that LC3 protein was mainly located in cytoplasm.The expression of LC3 in model group and experimental group were significantly higher than that in blank group,and the expression level of LC3 in experimental group was higher than that in model group.oα-synuclein protein was distributed evenly in the cytoplasm and nucleus.The expression of α-synuclein in model group and experimental group was higher than that in blank group,experimental group was lower than that in model group.Conclusion In Parkinson' s disease cell model,Nur77 agonist can promote the degradation of α-synuclein,the mechanism may be related to increase autophagy level.

Objective To investigate the feasibility of retroperitoneoscopy in the treatment of adrenal masses in infants under 6 months old.Methods From January 2020 to November 2023,retroperitoneoscopic surgery was performed in 5 infants under 6 months old with adrenal tumors.Their age was from 1 month and 18 days to 4 months and 27 days,and their body weight was 5-8 kg.The lesion was found by prenatal ultrasonography in 1 case and by abdominal ultrasonography for other reasons after birth in 4 cases.Ultrasound and CT indicated a diameter of 1.7-5.5 cm for the adrenal masses.Results The operations of adrenalectomy and tumor resection were completed under retroperitoneoscopy.The operative time was 65-135 min(median,94 min).The intraoperative blood loss was less than 10 ml.The postoperative drainage tube retention time was 3-6 d(median,5 d).Pathological diagnosis showed 4 cases of adrenal neuroblastoma and 1 case of adrenal hyperplasia.Follow-ups for 1-36 months(median,3 months)with abdominal ultrasound and CT scanning showed no recurrence or metastasis.Conclusion Retroperitoneoscopy is relatively safe for the treatment of adrenal tumors in infants under 6 months old(tumors with acceptable boundaries).

Objective To summarize the clinical experience of thoracoscopic treatment of anterior mediastinal enterogenous cysts.Methods From July 2019 to July 2023,27 children diagnosed as having anterior mediastinal enterogenous cysts were treated with total thoracoscopic surgery through the lateral thoracic approach(three port method)in our department.The patients were placed in a healthy lateral position.The observation hole was located in the 5th intercostal space under the scapula,and the other 2 operating holes were established based on the location of the lesion and the endoscopic diamond-shaped method,both of which were 5 mm trocars.The CO2 pneumothorax was established at a pressure of 6 mm Hg.The visceral pleura of the cyst was opened with an electric hook,the cyst was fixed and pulled by intestinal forceps,and the cyst was completely removed by forceps and electric hook separation alternately.Results No conversion to thoracotomy was required.Complete resection was performed in 26 cases,and residual cyst wall existed in 1 case.Esophageal muscular layer was opened in 9 cases.During the operation,cysts obstructed the surgical field of view in 5 cases,which was not conducive to observation.The cyst puncture and fluid extraction were performed.The operation time was 45-120 min(median,70 min).The amount of blood loss was 3-10 ml(median,5 ml).Postoperative hospitalization lasted for 2-5 d(median,3d).The 27 cases were followed up for 1-43 months(median,22 months),and there was no recurrence.The compressed trachea in 2 cases was all recovered,with emphysema fully recovered.Conclusions Thoracoscopic treatment of anterior mediastinal enterogenous cysts in children is safe and feasible.When the surrounding structure of the lesion is complex,it can be combined with bronchoscopic or gastroscopic surgery if necessary.