Loss of ATM activity induces GADD45α-dependent apoptosis in cerebellar granular neurons / 中山大学学报(医学科学版)

objectiveTo explore the specific molecular mechanism of neuronal apoptosis induced by ATM inactivation. MethodsCGNs matured 7 days in vitro were cultured 8 h with 25 K， 5 K or 25 K medium containing ATM-specific inhibitors （Ku55933， 10 µmol/L； Ku60019， 15 µmol/L） for Hoechst stain and apoptosis analysis， or cultured for different lengths of time （2， 4， 8 h） to detect the protein expression levels of ATM， caspase-3 and cleaved caspase-3 by Western blotting. ATM and GADD45α specific siRNA was transfected into C6 cells and CGNs， and its interference efficiency was verified by q-PCR and Western blotting. CGNs matured for 5 days in vitro were transfected with ATM specific siRNA and pCMV-EGFP by calcium phosphate for 48 h， Hoechst staining and apoptosis analysis were performed. CGNs matured for 7 days in vitro were treated with 25 K medium containing ATM specific inhibitors for 8 h， transcriptome sequencing， differential expression gene identification and pathway enrichment analysis were performed. CGNs matured for 5 days in vitro were co-transfected with GADD45α specific siRNA and pCMV-EGFP by calcium phosphate for 48 h， then treated with 5 K or 25 K medium containing 15 µmol/L Ku6 for 8 h. Hoechst staining and apoptosis analysis were performed. ResultsCompared with the 25 K， CGNs nuclear pyknosis rate， cleaved Caspase-3 and ATM protein expression level were increased in the 5 K and ATM-specific inhibitor groups. The mRNA and protein expression levels of ATM and GADD45α were effectively reduced after transfection of ATM and GADD45α specific siRNA in C6 cells and CGNs. Compared with control， CGNs transfected with ATM specific siRNA showed a higher nuclear pyknosis rate. Totally 835 genes were identified to be up-regulated and 848 genes to be down-regulated in the Ku55933 treatment group； 454 genes were identified to be up-regulated and 314 genes to be down-regulated in the Ku6 treatment group； 274 genes were co-up regulated in the Ku5 and Ku60019 treatment groups， while 179 genes were co-down-regulated in the Ku5 and Ku6 treatment groups and the expression of ATM downstream target GADD45α was upregulated. The enrichment results showed that TNF signaling pathway， NF-κB signaling pathway and Apoptosis signaling pathway were significantly enriched. Compared with control， mRNA and protein expression levels of GADD45α were increased in inhibitor treatment and 5 K， while knocking down GADD45α resulted in a decrease in nuclear pyknosis rate in the Ku60019 and 5 K treatment group. ConclusionLoss of ATM activity induces GADD45α-dependent cerebellar granular neuronal apoptosis.

Three-dimensional finite element analysis on stress distribution after greater trochanter bone flap for treating osteonecrosis of the femoral head / 中国组织工程研究

BACKGROUND: Three-dimensional finite element analysis has been used by many scholars from department of orthopedics, but the results of postoperative evaluation of hip preserving treatment for osteonecrosis of femoral head are different. OBJECTIVE: To study the biomechanical changes of the femoral head and the biomechanical changes of the proximal femur after greater trochanter bone flap for the treatment of femoral head necrosis using three-dimensional finite element method, and to verify the mechanical safety and effectiveness. METHODS: One case of unilateral femoral head necrosis in ARCOIII stage undergoing parallel vascularized greater trochanter bone flap transplantation was selected. Computed Tomography data of proximal femur were collected before and 6 months after the operation, and preserved in DICOM format. With the aid of computer technology, professional medical modeling software, MIMICS and HYPERMESH, were used to establish the three-dimensional geometric models of the proximal femur. These models were divided into normal group, necrosis group and repair group. Finite element analysis software ANSYS was utilized to simulate human body standing and movement in different situations. The model was divided by free mesh, and given material parameters to establish normal proximal femur, femoral head necrosis and bone defect. Greater trochanter bone flap was applied in repairing three-dimensional finite element model of bone defect. Loads were loaded on different finite element models. The maximum displacement of the femoral head and the stress distribution in the proximal femur of the three groups were observed under different loading models. RESULTS AND CONCLUSION: (1) Under the same load, the maximum displacement of the three sets of models was 0.61 mm in the normal group, 0.66 mm in the necrosis group, and 0.61 mm in the repair group, respectively. Maximum Von Mises stress was greater in necrosis model than in the normal molding. The maximum Von Mises stress gradually decreased in the repair model, and was close to normal value. (2) Three groups of models showed stress concentration above the rotor in femoral neck region. The maximum stress in the trochanteric position was higher in necrosis models than in normal models. The maximum stress in this region gradually increased after repair, but was still lower than the failure stress of bone. (3) The results confirm that the maximum stress and the maximum displacement are closer to the normal value after greater trochanter bone flap for treatment of osteonecrosis of the femoral head. The greater trochanter is safe and reliable for repairing bone defect of femoral head.

The clinical engineering strategy for reducing application risk of ventilator / 中国医学装备

Objective: To reduce the risk of ventilator in clinical use by using technical management so as to enhance cure rate for patients. Methods: Through a series of methods included of clinical investigation, statistic analysis of data, socio-technical system model and others to analyze and assess risk factors which were possibly produced in the cure process of ventilator. And the relevant risk regulation indicators were formulated as above information. Results: The formulation about risk control criterion of operation in the cure process of ventilator provided various aspects, which included of the material qualities of ventilator and breathe pipeline, clinical service environment, human factor and others, for clinical engineer to control risk so as to ensure the life safe of patients. Conclusion: The clinical engineering department should adopt technique management way to analyze the cure process of ventilator, and control risk from aspect of clinical engineering. Therefore, it can efficiently reduce the risk of ventilator in clinical application and promote the safety and efficiency of ventilator in the treatment.

Synergistic Killing Effects of Arsenic Trioxide Combined with Itraconazole on KG1a Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1a cells and its potential mechanism.</p><p><b>METHODS</b>The cell morphology was observed with Wrighe-Giemsa staining; cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2,caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot.</p><p><b>RESULTS</b>The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group, both the cell survival rate and the colony number of the single drug-treatment group were significantly lower(P<0.05), and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group, the protein expression of Caspase-3 and BAX was upregulated, while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated.</p><p><b>CONCLUSION</b>Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis, which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression, and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.</p>

Proteomic Analysis of the Peritrophic Matrix from the Midgut of Third Instar Larvae, Musca domestica / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To better comprehend the molecular structure and physiological function of the housefly larval peritrophic matrix (PM), a mass spectrometry approach was used to investigate the PM protein composition.</p><p><b>METHODS</b>The PM was dissected from the midgut of the third instar larvae, and protein extracted from the PM was evaluated using SDS-PAGE. A 1D-PAGE lane containing all protein bands was cut from top to bottom, the proteins in-gel trypsinised and analysed via shotgun liquid chromatography- tandem mass spectrometry (LC-MS/MS).</p><p><b>RESULTS</b>In total, 374 proteins, with molecular weights varying from 8.225 kD to 996.065 kD and isoelectric points ranging from 3.83 to 11.24 were successfully identified, most identified proteins were mainly related to immunity, digestion, nutrient metabolism and PM structure. Furthermore, many of these proteins were functionally associated with pattern binding, polysaccharide binding, structural constituent of peritrophic membrane and chitin binding, according to Gene Ontology annotation.</p><p><b>CONCLUSION</b>The PM protein composition, which provides a basis for further functional investigations of the identified proteins, will be useful for understanding the housefly larval gut immune system and may help to identify potential targets and exploit new bioinsecticides.</p>

A Comparison of Brain Death Criteria between China and the United States / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Criteria for determining brain death (BD) vary between China and the United States. We reported the results of an investigation designed to compare procedures to determine BD in two countries.</p><p><b>METHODS</b>The latest criteria in the United states were published in 2010. The latest criteria in China were published in 2009. We used these two types of BD criteria to evaluate patients who were considered to be BD. The time, cost, and accuracy of the diagnosis were compared.</p><p><b>RESULTS</b>From January 1, 2012 to October 8, 2013, there were 37 patients which were applied for BD evaluation in the Neurological Intensive Care Unit of Beijing Tiantan Hospital. The cause of coma were known as subarachnoid hemorrhage (18 patients, 48.6%), intracerebral hemorrhage (8 patients, 21.6%), cerebral ischemia (9 patients, 24.3%), brain stem tumor (1 patient, 2.7%), and intracranial infection (1 patient, 2.7%). The clinical examinations were done for all of the patients except 1 patient who had low blood pressure. Three patients had brainstem reflexes that were excluded from BD. Twenty-five patients had apnea tests, and 20 tests were completed that were all positive. Confirmatory tests were completed differently: Transcranial Doppler (30 patients, positive rate 86.7%), electroencephalogram (25 patients, positive rate 100%), and somatosensory evoked potential (16 patients, positive rate 100%). Thirty-three patients were diagnosed BD by criteria of the United States. Only 9 patients were diagnosed BD by Chinese criteria. The use of time and money in the USA criteria was obviously fewer than those in Chinese criteria (P = 0.000).</p><p><b>CONCLUSION</b>Compared with BD criteria of the United States, Chinese criteria were stricter, lower positive rate, more cost in money and time, and more reliable by families and doctors.</p>

Case control study on therapeutic effects of dynamic external fixtor combined with limited internal fixation and cross K-wires fixation for the treatment of Pilon fractures of the proximal interphalangeal joint / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effects and safety of dynamic external fixtor combined with limited internal fixation and cross K-wires fixation for the treatment of close Pilon fractures of the proximal interphalangeal joint.</p><p><b>METHODS</b>From June 2012 to June 2014, totally 41 patients (45 fingers) with close interphalangeal joint Pilon fracture were treated by dynamic external fixtor combined with limited internal fixation or cross K-wires fixation, and all the patients were followed up. In the dynamic external fixtor combined with limited internal fixation group (group A), there were 21 patients with 22 fingers, including 12 males and 9 females, with an average of (30.6±5.6) years old. In the cross K-wires fixation group (group B), there were 20 patients with 23 fingers, including 11 males and 9 females, with an average of (30.1±5.3) years old. Regular re-examination of X-ray was performed to evaluate the active range of joint motion, fracture healing time, infection rate and postoperative joint motion pain.</p><p><b>RESULTS</b>According to the evaluation criteria of upper extremity function issued by the Hand Surgery Society of Chinese Medical Association, the excellent and good cases of group A was up to 19 and 13 for group B. The evaluation results has significant differences (Z=2.558, P=0.011). The excellent and good rate of group A was obviously higher than that of group B. The average bone union time of group A was (7.9±2.1) weeks, and (8.1±2.3) weeks for group B. There was no significant difference on the mean healing time (t=-0.304, P=0.762). The infection fingers of group A was 5, and 1 for group B. The difference between the results was statistically significant (χ2=3.287, P<0.05). The infection rate of group A was higher than that of group B. The postoperative joint motion pain was evaluated by VAS score, the mean score was 0.18±0.50 in group A, and 0.65±0.88 in group B. The difference between the results was statistically significant (t=-2.207, P<0.05). The postoperative joint motion pain was lower than that of group B.</p><p><b>CONCLUSION</b>Dynamic external fixtor combined with limited internal fixation is a reliable and effective method to treat Pilon fractures of the proximal interphalangeal joint. It allows early postoperative functional rehabilitation and restores the joint function.</p>

Treatment of chronic mallet finger deformity with minor bone anchors and palmaris longus tendon graft / 中国骨伤

<p><b>OBJECTIVE</b>To explore the clinical effects of minor bone anchors and palmaris longus tendon graft in treating chronic mallet fingers deformity.</p><p><b>METHODS</b>From January 2008 to June 2013, 26 patients with chronic mallet fingers deformity were treated with minor bone anchors and palmaris longus tendon graft. There were 18 males and 8 females, aged from 18 to 52 years old with an average of (32.0±1.3) years. Among them, 8 cases caused by machine injury, 6 cases by fall injury, 6 cases by sprain from fight, 4 cases by tendon spontaneous rupture, 2 cases by knife trauma. There was no tendon attachment of extensor tendon check in 16 cases, and with 0.3 to 0.5 cm tendon attachment in 10 cases. All patients had the flexion deformity and the disability of dorsiflexion activity. During operation, the distal interphalangeal joint was fixed in 10° to 20° dorsiflexion by a Kirshner wire, the minor bone anchor was used to reconstruct the extensor tendon insertion, the palmaris longus tendon slice was transplanted the decayed area of extensor tendon insertion. Four weeks postoperatively, the Kirshner wire was removed and the plaster external fixation was used, and the patient began function exercises. Postoperative complications were observed and fingers functions were assessed according to Dargan standard.</p><p><b>RESULTS</b>The patients were followed up from 6 to 14 months with an average of (5.0±0.3) months. Wound superficial infection occurred in 2 cases, the skin pressure ulcer in 2 cases, joint activities disability in 1 case; these symptoms got improvement after symptomatic treatment. Traumatic arthritis occurred in 2 cases, 1 case was improved after treatment, and 1 case had chronic pain for a long time. No internal fixation loosening or breakage and tendon rupture were found. According to Dargan standard to evaluate the finger function, 17 cases got excellent results, 8 good, and 1 poor.</p><p><b>CONCLUSION</b>It is an effective way to treat the chronic mallet finger deformity using minor bone anchors and palmaris longus tendon graft, and the method has advantages of reliable fixation, easy operation, satisfactory effect and less complication.</p>

Synergistic killing effect of arsenic trioxide combined with curcumin on KG1a cells / 中国实验血液学杂志

This study was aimed to explore the effect of arsenic trioxide combined with curcumin on proliferation and apoptosis of KG1a cells and its potential mechanism. The cell survival rate was mesured by MTT; colony formation capacity was examined by methylcellulose colony formation test; flow cytometry was used to analyse the cell surface molecules, cell apoptosis rate and cell cycle; the cell morphology was observed with Wright-Giemsa staining and the protein expression of BCL-2, BAX, PARP was detected by Western blot. The results showed that the phenotype of KG1a cells was CD34(+)CD38(-), while the phenotype of HL-60 cell was CD34(+)CD38(+). The former possessed a stronger colony ability than the latter. Effect of curcumin and arsenic trioxide alone on cell proliferation and inhibition was in dose-dependent manner. Compared with single drug-treatment group, the cell survival rate and colony number were lower, and the apoptosis rate was higher in combined drug-treatment group. Protein expression of BCL-2 and PARP was upregulated, while the protein expression of PARP was downregulated in the combined treatment group. It is concluded that compared with HL-60 cells, KG1a cells are the earlier leukemia stem/progenitor cells. Arsenic trioxide combined with curcumin can effectively inhibit the KG1a cell proliferation and induce apoptosis, which may be associated with the downregulation of BCL-2 and PARP protein expression and the upregulation of BAX protein expression.

Effect of honokiol on proliferation and apoptosis in HL-60 cells and its potential mechanism / 中国实验血液学杂志

This study was aimed to investigate the effect of Honokiol (HNK) on proliferation and apoptosis of acute myeloid leukemia HL-60 cells and its potential mechanism. Inhibitory effect of HNK on the HL-60 cell proliferation was detected by MTT assay. Flow cytometry was used to detect the change of cell cycle and AnnexinV/PI staining was used to detect apoptosis. Western blot was applied to analyze the cell cycle protein (cyclins), cyclin-dependent kinase (CDK), P53, P21, P27, BCL-2, BCL-XL, Bax, caspase-3/9 and proteins for MAPK signal pathway. The results showed that HNK could inhibit the proliferation of HL-60 cells in time- and dose dependent ways. HNK arrested HL-60 cells in G0/G1 phase, and S phase cells decreased significantly (P < 0.05). The expression of cyclin D1, cyclin A, cyclin E and CDK2/4/6 were significantly down-regulated (P < 0.05), the expression of P53 and P21 was significantly upregulated after treating for 24 h with HNK (P < 0.05). After 24 h treatment with HNK, HL-60 cell apoptosis increased significantly with the upregulation of activated caspase-3, -9, BAX expression and the downregulation of BCL-2, BCL-XL expression. The MAPK subfamily, P38 and JNK were not significantly changed, but the expression of MEK1/2-ERK1/2 was significantly downregulated (P < 0.05). It is concluded that HNK arrestes the cells at G0/G1 phase and induces HL-60 cell apoptosis through the intervention of MEK1/2-ERK1/2 signaling pathway.

Effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels in a rabbit model of lung ischemia-reperfusion injury / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels during lung ischemia-reperfusion injury in rabbits.</p><p><b>METHODS</b>Eighteen healthy rabbits were randomly divided into three groups: control group (group C, n = 6), ischemia/reperfusion group (group I/R, n = 6), limb ischemia preconditioning group (group L, n = 6). At the end of experiments, the wet to dry-weight ratio (W/D), activities of superoxide dismutase (SOD) and myeloperoxidase (MPO), levels of malondialdehyde (MDA) and the contents of cytokines (TNF-α, IL-6, IL-8 and IL-10) were determined in lung tissues. Protein levels of bronchoalveolar lavage fluid and serum were measured to calculate the lung permeability index. Pathologic changes of lung tissues were also observed.</p><p><b>RESULTS</b>Compared to the group I/R, the lung tissue W/D ratio, MPO activity, lung permeability index, MDA and the cytokines (TNF-α, IL-6 and IL-8) levels were significantly decreased in group L (P < 0.05), while the SOD activity (P < 0.05) and IL-10 contents were significantly increased (P < 0.01). There was no statistical difference in the changes of the above parameters between group L and group C (P > 0.05). The morphologic damages were significantly reduced in group L than that in group I/R.</p><p><b>CONCLUSION</b>Limb ischemia preconditioning has protective effect against lung ischemia-reperfusion injury, which may at least in part through inhibiting the release of oxygen-derived free radicals and pro-inflammatory cytokines (TNF-α, IL-6, IL-8) and increasing the production of anti-inflammatory cytokine IL-10.</p>

Pattern of electrogastrogram in healthy neonates / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the electrogastrogram (EGG) characteristics of healthy neonates.</p><p><b>METHODS</b>Twenty healthy neonates born at 37-39 weeks of gestation (11 males and 9 females, Apagar's score 9.3 +/- 0.4) were enrolled in this study. EGG recordings were performed for half an hour pre- and postprandially at an interval of a week from birth until age 4 weeks. The EEG variables measured included the percentage of normal gastric rhythm, the percentage of tachygastria and bradygastria, the fed-to-fasting ratio of the EEG dominant power, as well as the EEG dominant frequency and its instability coefficient. The paired sample t test (95% CI) was used to compare the recordings.</p><p><b>RESULTS</b>Between birth and age 28 days, the percentage of normal gastric rhythm ranged from 38.2 +/- 4.9% to 39.7 +/- 3.5% of recorded time, tachygastria was observed in the range of 23.7 +/- 5.4% to 23.5 +/- 4.3% of recorded time, and bradygastria was shown to be in the range of 38.1 +/- 5.5% to 36.8 +/- 3.9% of recorded time in the 20 neonates before meal. Statistically significant differences were not seen in neonates with different ages as well as during pre- and postprandial periods. The EEG dominant frequency of neonates before meal was 2.38 +/- 0.5, 2.43 +/- 0.2, 2.54 +/- 0.3, 2.57 +/- 0.2 and 2.59 +/- 0.1 cpm at birth and at postnatal age of 7, 14, 21 and 28 days respectively. There were no significant differences in the dominant frequency and the coefficient of instability of the dominant frequency during pre- and postprandial periods. The EEG dominant frequency at postnatal age of 14, 21 and 28 days during pre- and postprandial periods was significantly higher than that at birth and at postnatal age of 7 days (P < 0.05). The coefficient of instability of the dominant frequency at postnatal age of 21 and 28 days was significantly lower than that at birth and at postnatal age of 7 and 14 days (P < 0.05). There were no statistically significant differences in the fed-to-fasting ratio of EGG dominant power in neonates with different ages.</p><p><b>CONCLUSIONS</b>The pattern of electrical activity in the normal neonatal stomach appears to be different from that demonstrated in adults and children. The percentage of normal gastric rhythm is lower, and tachygastria and bradygastria are more frequently seen. The EEG dominant frequency increases with postnatal age in neonates.</p>

Study on the prevalence rate of rheumatoid arthritis in Zhuang nationality population from Nanning, Guangxi / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi.</p><p><b>METHODS</b>A total of 7407 people with Zhuang nationality, aged 16 and over were surveyed. Questionnaire was administered to each subject under survey, while X-ray and serum rheumatoid factor were carried out to all those who gave positive answers. Physical examinations were done to the suspicious cases by experts on rheumatism.</p><p><b>RESULTS</b>were compared with those in 6826 people of Han ethnicity living in the same district.</p><p><b>RESULTS</b>The prevalence rates of rheumatoid arthritis in Zhuang nationality population was 0.27% when comparing to the Han population which was 0.28% (P > 0.05). Rates of awareness on rheumatoid arthritis in Zhuang and Han population were 5.0% and 10.5% (P > 0.05). After the diagnosis of rheumatoid arthritis was made and among patients who had received the treatment, the rates were 0% vs. 5.25%.</p><p><b>CONCLUSION</b>The prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi was not significantly different than that in Han ethnic group. However, the rates on awareness and the treatment of rheumatoid arthritis were still under satisfaction.</p>

Detection of hantaan virus from gamasid mite and chigger mite by molecular biological methods / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the proliferation and location of hantaan virus (HV) in gamasid mites and chigger mites.</p><p><b>METHODS</b>HV RNA in gamasid mites and chigger mites were detected by reverse transcription, polymerase chain reaction (RT- PCR) and in situ hybridization.</p><p><b>RESULTS</b>The smallest quantity of mite from which HV RNA could be detected was 5 mites group. The titers of -and proliferated in mites HV RNA could be found in ovary cells and dug cells of gamasid mites and chigger mites by in situ hybridization.</p><p><b>CONCLUSIONS</b>The results showed that HV could be trans-stadially transmitted and proliferated in mites, and HV always located in ovary and dug organs of mites. These results provide direct evidence at molecular level for the role of gamasid mites and chigger mites as vectors in transmission of HV.</p>