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Objective:To summarize the clinical manifestations of a case of combined oxidative phosphorylation deficiency 28 (COXPD-28) caused by the mutations of the SLC25A26 gene, thus providing references for the diagnosis and genetic counseling of the disease. Methods:Clinical data of a case of COXPD-28 treated in the Third Affiliated Hospital of Zhengzhou University in October 2020 were retrospectively analyzed.In addition, The retrieval words " Combined oxidative phosphorylation deficiency 28, SLC25A26 gene" were used to search domestic and foreign databases.The clinical characteristics of combined phosphorylation deficiency 28 and the variation characteristics of SLC25A26 gene were summarized. Results:(1) A female patient full-term delivered after 30 min presented with groaning breath was admitted.Her main manifestations included pale complexion, groaning breathing, metabolic acidosis, and high lactate and pyruvate levels.Symptomatic support treatment like anti-infection and assisted ventilation were given, but her condition gradually worsened and died of respiratory and circulatory failure on the day of admission.The child was compound heterozygous mutation of SLC25A26 gene, the terminating mutation of exon 5 c. 403G>T caused the protein change to p. E135, and the non-synonymous mutation of exon 4 c. 212A>G caused the protein change to p. Y71C.(2) A total of 3 cases of COXPD-28 were searched in online databases, and no cases were reported in China.Through literature review, clinical features of COXPD-28 mainly included respiratory and circulatory fai-lure, elevations of lactate and pyruvate, and reductions of complexes Ⅰ, Ⅲ and Ⅳ in muscle biopsy.Two types of mutations in the SLC25A26 gene were detected, including 3 cases of missense mutations and 1 case of splicing mutation. Conclusions:COXPD-28 is an autosomal recessive genetic disease involving multiple systems and mitochondrial dysfunction.Mutations in the SLC25A26 gene is the pathological cause of COXPD-28.
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Objective To study the effect of the inhibitor of apoptosis protein,Livin on proliferation and multi?drug resistance of lung adenocarcino?ma cells A549. Methods A549 cells were transfected with the eukaryotic expression vector pcDNA3.1?Livin. A549 cell clone with stable expres?sion of Livin was obtained through G418 screening. Expressions of Livin mRNA and protein in the transfected cells were respectively measured by re?verse transcription polymerase chain reaction(RT?PCR)and Western blot. The distribution of cell cycle phase was determined using flow cytometry. The level of P?gp mRNA and protein in A549 cells transfected with pcDNA3.1?Livin was detected by RT?PCR and Western blot. The analysis of multi?drug resistance of A549 treated with different chemotherapeutics was performed by MTT. Results The mRNA and protein expressions of Liv?in were both significantly increased in the transfected A549 cells. The flow cytometry analysis showed there was higher percentage of S phase and low?er percentage of G0/G1 phase in A549 cells transfected with pcDNA3.1?Livin. Compared with control groups,the expression of P?gp mRNA and pro?tein was increased in A549 cells transfected with pcDNA3.1?Livin,which showed a higher drug resistance and lower sensitivity to chemotherapic drugs such as ADM,MTX,CTX,and DDP(P<0.05). Conclusion Overexpression of Livin could enhance the proliferation of A549 cells,and high expression of P?gp caused by Livin could serve as one of the causes for multi?drug resistance in lung adenocarcinoma against chemotherapies.
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Objective To investigate the effects of small interfering RNA (siRNA) silencing Nanog gene on the ability of migration and invasion of the human lung adenocarcinoma A549 cells. Methods The human lung adenocarcinoma A549 cells were transfected with siRNA targeting Nanog gene , and three experiment groups were set up. The expression level of Nanog was detected using reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Cell migration was examined by wound healing assay and cell invasion was detected by Transwell assay. Results The Nanog silencing cell group (A549-siNanog) showed much lower level of Nanog mRNA and protein (0.40 ± 0.06, 0.50 ± 0.03) than A549-siNC cell group (0.97 ± 0.03, 0.85 ± 0.02; P < 0.05) under RT-PCR and Western blot analysis. Meanwhile, the wounded area filled rate and the number of invaded cells of A549-siNanog cell group (57% ± 0.04, 69.60 ± 17.14) were decreased significantly compared to A549-siNC cell group (95% ± 0.02, 209.60 ± 15.40; P < 0.05). Conclusion siRNA targeting human Nanog could specially suppress the expression of Nanog gene in lung adenocarcinoma A549 cells. In this way, it couldsignificantly reduce the capability of migration and invasion of A549 cells.
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BACKGROUND:In recent years, monitoring the pressure in the human body, especial y esophageal variceal pressure, becomes a hot spot. A lot of progress has been achieved regarding fiber optical sensors for measurement of the pressure in the human body. OBJECTIVE:To briefly review the fiber optical sensor applications in the human body. METHODS:A computer-based online retrieval was performed to search papers in CNKI periodical ful-text database and PubMed database (from January 1983 to March 2013) using the key words of“fiber optical sensor, pressure, measurement”in Chinese and English, respectively. After excluding objective-independent and repetitive papers, 40 papers were included for further analysis. RESULTS AND CONCLUSION:Compared with traditional sensors, fiber optical sensors, which have advantages in high sensitivity, large dynamic range, fast response, tolerance to electronic interference, explosion proofing, fireproofing and corrosion protection, have been used to measure esophageal variceal pressure, intracranial pressure, pharyngeal pressure, pediatric airway pressure, cardiovascular&blood pressure, intervertebral disc pressure, intrauterin pressure in childbirth, pressure in the colon, plantar pressure and shear force as wel as other pressures in the human body. Fiber optical sensors have been used more widely in pressure monitoring. With the development of production technology and device performance, fiber optical sensors wil further promote the rapid development of medical science in the near future.
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@#Objective To investigate the effects of RNA interference (RNAi) on Caspase-3 expression.Methods According to the gene sequence and the secondary structure characteristics of Caspase-3, three siRNA （Si-Caspase-3-1, Si-Caspase-3-2 and Si-Caspase-3-3） were designed with 21 basic group, and siRNAs against Caspase-3 gene were synthesized. With the liposome conduct, the synthesized siRNAs were transferred into neuroglia cells. After the cells crept the slices, cells were dyed with fluor Hoechst33258; the apoptotic cells were stained with TUNEL, the normal neuroglia cells were as control, the Caspase-3 protease activity were detected with hemi-quantitative RT-PCR and Western blot.Results The synthesized siRNAs could inhibit the expression of Caspase-3 gene. For Si-Caspase-3-3, the expression inhibition rate of Caspase-3 mRNA was 86.32% in neuroglia cell.Conclusion The synthesized siRNAs can inhibit the expression of Caspase-3 gene at cell level.
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Objective To study the expression of Livin and its relalionship with expression of p53 in small cell hmg cancer(SCLC).Methods Immunohistochemical S-P method was used to detect the ex-pression of Livin and p53 protein in 30 SCLC tissues and 16 para-cancerous lung tissues.Results Livin protein was expressed in 17 of 30 SCLC tissues(56.67%),but Livin protein showed low levels in para-can-cerous lung tissues(12.50%),P＜0.01.There was no significant correlalion between positive Livin protein expression and age,sex,TNM staging,lymph node metastasis and lumor diameter(P＞0.05).p53 protein was expressed in 14 of 30 SCLC tissues(46.67%),but p53 protein was no expressed in para-caneerous lung tissues,P＜0.01.The expression of Livin protein was positively related to the expression of p53 protein(P＜0.01).Conclusions The aberrant expression of Livin may be a new target for diagnosis and gene treatment of SCLC.The aberrant expression of Livin and p53 may play synergetic role in process of carcinogenesis of SCLC.
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To study the expression of neurocyte apoptosis and the changes of caspase-3 and Fas after spinal cord injury (SCI) in rats, improved Allen's method was used to make model of acute SCI at the level of T9 and T10. The animals were divided into six groups: a control group and 5 injury groups. The segments of injured spinal cords were taken 6, 24, 48 h and 7, 15 days after injury for morphological studies, including HE staining, Hoechst33258 staining and TUNEL labeling. The expression of caspase-3 was detected by immunohistochemical staining and RT-PCR. TUNEL-positive cells began to appear in the compression region 6 h after the injury, mostly located in the gray matter. TUNEL-positive cells were found in both gray and white matter, reaching a peak at the 3rd day. They began to decrease at the 7th day, distributed mostly in the white matter. Fas increased at the 6th h and peaked at the 3th day. Caspase-3 mRNA increased at the 6th h, peaking 48 h after the trauma, and decreased after 7 days. The protein expression of caspase-3, as revealed by immunohistochemical staining, was similar to TUNEL in time. It is concluded that apoptosis takes place after spinal cord injury, and caspase-3 mRNA and protein expressions were enhanced in the apoptosis. The expression of caspase-3 has a positive correlation with Fas expression.
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<p><b>BACKGROUND</b>The expressive level of glucose-regulated protein 78 (GRP78) is elevated and correlated with resistance of chemotherapy drugs in breast cancer cell. However, little is known about the relationship between its expression and drug resistance in non-small cell lung cancer (NSCLC). The aim of this study was to explore the relationship between drug resistance and the expression of GRP78 in NSCLC.</p><p><b>METHODS</b>Drug sensitivity test was used to detect the resistance to 8 chemotherapy drugs in 52 NSCLC fresh surgical samples by methylthiazoletrazolium (MTT), and expression of GRP78 was detected by immunohistochemistry method. Spearman correlation assay was used to investigate the correlation between the GRP78 expression and drug resistance.</p><p><b>RESULTS</b>The resistance rates to paclitaxel (PTX), adriamycin (ADM), carboplatin (CBP), topotecan (TPT), navelbine (NVB), vincristine (VCR), cisplatin (DDP) and etoposide (VP-16) of the 52 samples were 42.31%, 57.69%, 63.46%, 65.38%, 67.31%, 73.08%, 78.85%, 90.38%, respectively. Fourteen cases showed the complete resistance to the total 8 chemotherapy drugs. Furthermore, the expression of GRP78 was stronger in poorly differentiated cancer as compared with the well and moderately differentiated cancer (P < 0.05), so as in stage II and III cancer than in stage I cancer (P < 0.05). Spearman correlation assay showed that there was a correlation between the chemotherapeutics resistance to ADM, VP-16, VCR, TPT and the expression of GRP78 in NSCLC (P < 0.05).</p><p><b>CONCLUSIONS</b>It is feasible to detect the drug sensitivity to chemotherapy for tumor cells by MTT method. The results of chemosensitivity assay in vitro are indicative of clinical drug administration in NSCLC. The detection of GRP78 isalso indicative of the resistance to chemotherapy drugs and the differentiation and the clinical stage in NSCLC.</p>
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To study the expression of neurocyte apoptosis and the changes of caspase-3 and Fas after spinal cord injury (SCI) in rats, improved Allen's method was used to make model of acute SCI at the level of T9 and T10. The animals were divided into six groups: a control group and 5 injury groups.The segments of injured spinal cords were taken 6, 24, 48 h and 7, 15 days after injury for morphological studies, including HE staining, Hoechst33258 staining and TUNEL labeling. The expression of caspase-3 was detected by immunohistochemical staining and RT-PCR. TUNEL-positive cells began to appear in the compression region 6 h after the injury, mostly located in the gray matter.TUNEL-positive cells were found in both gray and white matter, reaching a peak at the 3rd day. They began to decrease at the 7th day, distributed mostly in the white matter. Fas increased at the 6th h and peaked at the 3th day. Caspase-3 mRNA increased at the 6th h, peaking 48 h after the trauma, and decreased after 7 days. The protein expression of caspase-3, as revealed by immunohistochemical staining, was similar to TUNEL in time. It is concluded that apoptosis takes place after spinal cord injury, and caspase-3 mRNA and protein expressions were enhanced in the apoptosis. The expression of caspase-3 has a positive correlation with Fas expression.
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@# ObjectiveTo study the cognitive function and observe the changes of event-related potential of epileptic children.Methods45 epileptic children and 45 normal children were put in as the test group and the control group.The cognitive function and event-related potential were evaluated by means of Raven's standard Progressive Matrices (SPM) and evoked potential instrument respectively.ResultsAbout 33.3% epileptic children presented cognitive deficits.For children with cognitive deficits, their scores of B, E were significantly different from those in the control group(P<0.05).The latency of event-related potential P300 showed significantly prolonged than that of the control group (P<0.01).ConclusionEpilepsy may cause cognitive function deficit, especially in the ability of analog, analysis and abstract. The latency of event-related potential P300 is a very good objective to assess the cognitive function of children.
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@#ObjectiveTo observe the expression of apoptosis factor, the mechanism of neuronal apoptosis and profective effect of recombinant Human Erythropoietin (rHu EPO) after spinal cord injury (SCI) in SD rats.Methods30 SD rats were divided into four groups including control group, SCI group, treatment groups A and B.The animal SCI model was established with Allen's method. The changes of nerve functions of rats of 4 groups were observed before and after treating with rHu EPO. The expressions of apoptosis factors (Bcl-2,Bax,Fas) were tested with immunocytochemistry technique, and apoptosis neurones were labeled with TUNEL dyeing.ResultsThe grade of nerve function was improved distinctly in treatment groups, but group B was better than group A.The number of the positive cells for Fas and Bax in SCI group was more than that in treatment groups (P<0.05), and group A was more than group B. The number of Bcl 2 positive cells in the treatment groups was greater than that in SCI group (P<0.05).ConclusionApoptosis is a important death mode of neuron after SCI. rHu EPO can distinctly improve the comeback of the nerve function and protect the nerve tissue.
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To modify the surface property of poly lactide-co-glycolide (PLGA) by biomimetic mineralization to construct a new kind of artificial bone. PLGA films and 3-diamensional (3-D) porous scaffolds hydrolyzed in alkaline solution were minerilized in SBF for 14 days. The morphology and composition of the mineral grown on PLGA were analyzed with SEM, FTIR and XRD. The porosity of the scaffolds was detected by using the liquid displacement method. The compressive strength of the scaffolds was detected by using a Shimadzu universal mechanic tester. An obvious mineral coating was detected on the surface of films and scaffolds. The main component of the mineral was carbonated hydroxyapatite (HA) similar to the major mineral component of bone tissues. The porosity of the un-mineralized and mineralized porous scaffolds was (84.86 +/- 8.52) % and (79.70 +/- 7.70) % respectively. The compressive strength was 0.784 +/- 0.156 N/mm2 in un-mineralized 3-D porous PLGA and 0.858 +/- 0.145 N/mm2 in mineralized 3-D porous PLGA. There were no significant differences between the mineralized and un-mineralized scaffolds (P > 0.05) in porosity and biomechanics. Biomimetic mineralization is a suitable method to construct artificial bone.
Subject(s)
Biocompatible Materials , Bone Substitutes , Calcification, Physiologic , Durapatite/metabolism , Lactic Acid/chemistry , Polyglycolic Acid/chemistry , Polymers/chemistry , Porosity , Tissue EngineeringABSTRACT
BACKGROUND: Beside direct trauma, a series of secondary pathological changes would occur in local injured spinal cord area during spinal cord injury. It has been reported that recombinant human erythropoietin (rhEPO)could inhibit cell apoptosis and inflammatory reaction, and possess neuroprotective role.OBJECTIVE: To explore the neuroprotective role of rhEPO in spinal cord injury by observing the nerve cell apoptosis and related cytokine expression in traumatic spinal cord.DESIGN: Randomized and controlled study.SETTING: Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, and Department of Pathology of Tongji Medical College, Central China Science and Technology University.PARTICIPANTS: The study was conduced at Orthopedic Surgery Laboratory of Affiliated Cooperation Hospital of Tongji Medical College, Central China Science and Technology University and Department of Pathology of Tongji Medical College from September 2003 to May 2004. Thirty healthy female adult rats were randomly divided into 4 groups: ① Six rats in blank control group only subjected to spinal cord exposure without injury. ②Eight rats in injury group were subjected to spinal cord injury without medication. ③ Eight rats in medication A group were treated with rhEPO.④ Eight rats in medication B group were treated with thEPO and βaeacine sodium.METHODS: ① Animal model preparation: Spinal cord injury model was established on 24 rats by using improved Allen method. ② Administration: Rats in medication A group were treated with rhEPO in dosage of 300 U/(kg·d) at postoperative 1, 3, 5, 8, 11 days, while rats in medication B group were given additional β-aeacine sodium in dosage of 0.1 mg/kg,once a day for consecutive 11 days. Rats in blank control group and injury group were injected with the same volume physical saline from tail veins.③ Neurological function: The neurological function was scored by the same examiner at 1 day and 12 days after model establishment. Behavioral observation: Basing on improved Gale's neurological functional behavioral analysis, 0 score presented severer dysfunction and 6 scores represents normal. Slope test: The gradient was determined by the grasping capability of rat, which could reflect the recovery of neurological function. ④ Pathological examination: Rat was put to death at postoperative 12 days; traumatic spinal cord was made into slices for HE staining. ⑤ Expression of apoptosis cytokine bcl-2, bax and fas in nerve cells: Specimen was collected for IHC stain and brown colored positive cells was counted for calculating positive expressing rate. ⑥ Apoptosis nerve cells: In situ end-labeling techniques was used to calculate apoptosis index (the number of apoptosis nucleus/total number). All data were analyzed by using paired chi-test.MAIN OUTCOME MEASURES: ① Neurological functional behavioral score and results of slope test. ② Histological observation of traumatic spinal cord. ③ Expression of apoptosis nerve cell cytokines. ④ Examination of apoptostic nerve cells.RESULTS: ① Neurological functional behavioral score and results of slope test: There was no significant difference between 1 day and 12 days in blank control group, while the gradient in traumatic group was significantly larger in 12 days than in 1 day (P ＜ 0.05). In both therapeutic A and B groups, the behavioral scores and slope gradient were found significantly larger at 12 days than 1 day (P ＜ 0.05) and that of traumatic group (P ＜ 0.05). ② Histological observation of traumatic spinal cord: Traumatic spinal cord became slim with a majority of necrosis and glial cell hyperplasia in it; most of neurological tissues were found normal in medication A and B groups, with the neural structure of medication B group better than A group. ③Expression of nerve cell apoptosis cytokines: bax and fas positive cells in traumatic group were more than medication group A and B [traumatic group of (25.75±3.37)% and (41.37±2.83)% vs medication group A of (19.87±3.56) and (26.00±3.29)% vs medication group B of (12.00±2.97)and (17.50±2.20)％, P ＜ 0.05]; bcl-2 was significantly lower in medication group A and group B [(9.75±1.83)％, (14.63±2.83)％, (21.63±5.34)％,P ＜ 0.05]. ④ Examination of apoptotic nerve cells: Cell apeptosis index in traumatic group was significantly higher than medication group A and group B (50.75±5.39, 34.75±3.01, 24.00±3.46, P ＜ 0.05).CONCLUSION: Cell apoptosis is an important kind of neuronal death following spinal cord injury. rhEPO can inhibit nerve cell apoptosis and possess neuroprotective effect for traumatic spinal cord.
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To modify the surface property of poly lactide co-glycolide (PLGA) by biomimetic mineralization to construct a new kind of artificial bone. PLGA films and 3 diamensional (3-D) porous scaffolds hydrolyzed in alkaline solution were minerilized in SBF for 14 days. The morphology and composition of the mineral grown on PLGA were analyzed with SEM, FTIR and XRD. The porosity of the scaffolds was detected by using the liquid displacement method. The compressive strength of the scaffolds was detected by using a Shimadzu universal mechanic tester. An obvious mineral coating was detected on the surface of films and scaffolds. The main component of the mineral was carbonated hydroxyapatite (HA) similar to the major mineral component of bone tissues. The porosity of the un-mineralized and mineralized porous scaffolds was (84.86±8.52) % and (79.70±7.70) % respectively. The compressive strength was 0. 784±0. 156 N/mm2 in un-mineralized 3-D porous PLGA and 0. 858±0. 145 N/mm2 in mineralized 3-D porous PLGA. There were no significant differences between the mineralized and un-mineralized scaffolds (P＞0. 05) in porosity and biomechanics. Biomimetic mineralization is a suitable method to construct artificial bone.